Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Kiyoko Kato Last modified date:2022.06.16

Professor / Reproductive and Developmental Medicine / Department of Clinical Medicine / Faculty of Medical Sciences

1. Yahata H¹,Kodama K²,Okugawa K²,Hachisuga K²,Yasutake N²,Maenohara S²,Yagi H²,Yasunaga M²,Ohgami T²,Onoyama I²,Asanoma K²,Kobayashi H³,Sonoda K⁴,Baba S⁵,Ishigami K⁵,Ohishi Y⁶,Oda Y⁷,Kato K², Long-term follow up after sentinel node biopsy alone for early-stage cervical cancer, Gynecol Oncol, 10.1016/j.ygyno.2022.01.031, 165, 1, 149-154, 2022.04, Objective: Sentinel node biopsy alone (SNB) reduces the postoperative complications of pelvic lymphadenectomy, such as lymphedema and lymphangitis; however, the long-term prognosis after SNB is unclear. The objective of this study was to evaluate the long-term outcome and complications of patients with early-stage cervical cancer who underwent SNB for hysterectomy or trachelectomy.
Methods: We performed SNB for cervical cancer using a radioisotope method in 181 patients between 2009 and 2017. If the intraoperative sentinel lymph node evaluation was negative for metastasis, no further lymph nodes were removed.
Results: The median age of the patients was 34 years (range, 21-73 years). The International Federation of Gynecology and Obstetrics 2008 stage was IA1 in 6 patients, IA2 in 18, IB1 in 154, and IIA1 in 3. Of the 181 patients (44 with hysterectomy, 137 with trachelectomy), 8 did not undergo pelvic lymphadenectomy because of a false-negative intraoperative diagnosis, 20 received adjuvant therapy after surgery, and 4 (2.2%) experienced recurrence over a median follow-up period of 83.5 months (range, 25-145 months). In the four recurrent cases, recurrence occurred in the pelvis, lung, and bone in one patient each, while the remaining patient developed pelvic and para-aortic lymph node metastases. Of these four patients, one died, and the remaining three are alive without disease after multidisciplinary therapy. The 5-year progression-free and overall survival rates were 98.8% and 99.4%, respectively. Postoperative complications, such as lymphedema, were very low rate.
Conclusions: SNB for early-stage cervical cancer might be safe and effective, with no increase in the recurrence and postoperative complications rate.
Keywords: Cervical cancer; Long-term follow up; Sentinel node biopsy..
2. Nakahara K¹, Morokuma S², Maehara K¹, Okawa H¹, Funabiki Y³ ⁴, Kato K¹, Association of fetal eye movement density with sleeping and developmental problems in 1.5-year-old infants, Sci Rep, 10.1038/s41598-022-12330-1, 12, 1, 8236-8236, 2022.03, Eye movement density (EMD) is an evaluation index of rapid eye movements observed during sleep. This study aimed to investigate the association of fetal EMD with sleeping and developmental problems in infancy. We observed 60 normal singleton pregnancies (gestational age 28-37 weeks) using ultrasonography for 1 h. Fetal eye movements were counted, and EMD was calculated. Participants answered questionnaires regarding their child's sleep and development 1.5 years after their delivery. The outcomes of an infant's sleep were night awakening (yes or no), bedtime (before or after 22:00), and nighttime sleep duration (< 9 or ≥ 9 h). An infant's development was evaluated using the Child Behavior Checklist (CBCL) T-score. We found that decreased fetal EMD was associated with increased night awakening at the age of 1.5 years (odds ratio 0.84, 95% confidence interval 0.69-1.00 per unit decrease in EMD). However, fetal EMD was not associated with bedtime or nighttime sleep duration. In addition, fetal EMD was independently associated with the total problems T-score of the CBCL at the age of 1.5 years in the multivariate model (p = 0.047). In conclusion, fetal EMD may be associated with sleep and developmental problems in infants..
3. Kiyokoba R¹ ², Uchiumi T³ ⁴, Yagi M¹ ⁵, Toshima T¹, Tsukahara S¹, Fujita Y², Kato K², Kang D¹, Mitochondrial dysfunction-induced high hCG associated with development of fetal growth restriction and pre-eclampsia with fetal growth restriction, Sci Rep, 10.1038/s41598-022-07893-y, 12, 1, 4056-4056, 2022.03, Fetal growth restriction (FGR) and pre-eclampsia with fetal growth restriction (PE/FGR) are high-risk perinatal diseases that may involve high levels of human chorionic gonadotropin (hCG) and mitochondrial dysfunction. However, little is known about how these factors affect placental function. We investigated how mitochondrial dysfunction and high hCG expression affected placental function in unexplained FGR and PE/FGR. We observed elevated expression of hCGβ and growth differentiation factor 15 mRNA and protein levels in the placenta with both diseases. Likewise, antiangiogenic factors, such as Ang2, IP10, sFlt1, IL8, IL1B, and TNFα, were also upregulated at the mRNA level. In addition, the expression of COXI and COXII which encoded by mitochondrial DNA were significantly decreased in both diseases, suggesting that mitochondrial translation was impaired. Treatment with hCG increased Ang2, IP10, IL8, and TNFα mRNA levels in a dose-dependent manner via the p38 and JNK pathways. Mitochondrial translation inhibitors increased hCGβ expression through stabilization of HIF1α, and increased IL8 and TNFα mRNA expression. These results revealed that high expression of hCG due to mitochondrial translational dysfunction plays an important role in the pathogenesis of FGR and PE/FGR..
4. Morisaki N¹,Nagata C²,Morokuma S³,Nakahara K⁴,Kato K⁴,Sanefuji M⁵ ⁶,Shibata E⁷ ⁸,Tsuji M⁸ ⁹, Shimono M⁸ ¹⁰,Kawamoto T⁸,Ohga S⁵ ⁶,Kusuhara K⁸ ¹⁰, Japan Environment, Children’s Study Group, Lack of catch-up in weight gain may intermediate between pregnancies with hyperemesis gravidarum and reduced fetal growth: the Japan Environment and Children's Study, BMC Pregnancy Childbirth, 10.1186/s12884-022-04542-0, 22, 1, 199, 2022.03, Background: Women with nausea and vomiting of pregnancy (NVP) have higher birth weight infants, while those with hyperemesis gravidarum, a severe manifestation of NVP, have lower birth weight infants. We aimed to investigate the associations between maternal weight loss (a consequence of hyperemesis gravidarum), NVP, and infant birth weight.
Methods: This study was a secondary analysis of a nationwide birth cohort in Japan. Singleton pregnancies delivered at 28-41 weeks of gestation were included in the analysis. Women were categorized based on their weight change in the 1st trimester (as a proportion to their pre-pregnancy weight: > + 3%, > 0 to + 3%, > -3 to 0%, > -5 to -3%, ≤ -5%) and severity of NVP (no nausea, only nausea, vomiting but able to eat, vomiting and unable to eat). The effects of weight change and severity of NVP on infant birth weight and small for gestational age (SGA) were assessed using regression models. We further examined how these effects could be modified by maternal weight gain up to the 2nd trimester.
Results: Among 91,313 women, 5,196 (5.7%) lost ≥ 5% of their pre-pregnancy weight and 9,983 (10.9%) experienced vomiting and were unable to eat in the 1st trimester. Women with weight loss ≥ 5% in the 1st trimester had infants 66 (95% CI: 53, 78) g lighter and higher odds of SGA (aOR: 1.29; 95% CI: 1.14, 1.47) than women who gained > 3% during the same period. However, when adjusting for weight gain up to the 2nd trimester, women with weight loss ≥ 5% in the 1st trimester had infants 150 (95% CI: 135, 165) g heavier and lower odds of SGA (aOR: 0.39; 95% CI: 0.33, 0.46) than those who gained > 3% during the same period. In contrast, women with more severe NVP tended to have infants with larger birth weight and lower odds of SGA compared to women without NVP. These trends were strengthened when adjusting for weight gain up to the 2nd trimester.
Conclusions: Our study suggests the possibility that reduced fetal growth in pregnancies with hyperemesis gravidarum may be caused by the lack of catch-up in gestational weight gain up to the 2nd trimester.
Keywords: Birth weight; Fetal growth; Gestational weight gain; Hyperemesis gravidarum; Morning sickness; Nausea and vomiting of pregnancy; Small for gestational age..
5. Nakahara K¹, Michikawa T², Morokuma S³ ⁴, Hamada N¹ ⁴, Ogawa M¹ ⁴, Kato K¹ ⁴, Sanefuji M⁴ ⁵, Shibata E⁶ ⁷, Tsuji M⁶ ⁸, Shimono M⁶ ⁹, Kawamoto T⁶, Ohga S⁴ ⁵, Kusuhara K⁶ ⁹, The Japan Environment and Children's Study Group, Association of maternal hemoglobin levels during pregnancy with sleep and developmental problems in 1-year-old infants: A cohort study, Health Sci Rep, 10.1002/hsr2.552, 5, 2, e552, 2022.03, Background and aims: Maternal hemoglobin concentration during pregnancy is reported to be associated with various perinatal outcomes and may also be associated with infant development. This study aims to investigate the association between maternal hemoglobin levels during early or mid-pregnancy and sleep and developmental problems in 1-year-old infants.
Methods: We used the data of 66,935 pregnant women who were participants of the Japan Environment and Children's Study, a nationwide cohort study in Japan, between 2011 and 2014. Maternal hemoglobin level was examined at recruitment (mean gestational age, 15.3 weeks; SD, 2.85 weeks; range, 6-22 weeks). Information on infant sleep and development at the age of 1 year was acquired using a questionnaire. Infant development was evaluated using the Ages and Stages Questionnaire (ASQ).
Results: The mean (SD) maternal hemoglobin level was 12.0 (1.0) g/dl. Maternal hemoglobin levels were not associated with the majority of infant sleep and developmental outcomes. In the group with maternal hemoglobin <10.0 g/dl, the risk ratio (RR) for sleep at 22:00 or later was higher than that in the reference group with 11.0 g/dl ≤ hemoglobin < 14.0 g/dl (RR 1.12, 95% confidence interval = 1.00-1.25). In the analysis with maternal hemoglobin level as a continuous variable, both high and low hemoglobin levels were associated with a higher RR of a late bedtime. In addition, a low maternal hemoglobin level was associated with a higher RR for abnormal fine motor skills in the ASQ.
Conclusion: Our results suggest that a low level of maternal hemoglobin during pregnancy is associated with late bedtime and abnormal fine motor skills in 1-year-old infants. Conversely, a high level of maternal hemoglobin may also be associated with the infant's late bedtime.
Keywords: development; hemoglobin; infant; pregnancy; sleep..
6. Michikawa T¹, Morokuma S², Takeda Y¹, Yamazaki S³, Nakahara K⁴, Takami A⁵, Yoshino A⁵, Sugata S⁵, Saito S⁶, Hoshi J⁶, Kato K⁴, Nitta H³, Nishiwaki Y¹, Maternal exposure to fine particulate matter over the first trimester and umbilical cord insertion abnormalities, Int J Epidemiol, 10.1093/ije/dyab192, 51, 1, 191-201, 2022.02, Background: Our hypothesis was that exposure to fine particulate matter (PM2.5) is related to abnormal cord insertion, which is categorized as a form of placental implantation abnormality. We investigated the association between exposure to total PM2.5 and its chemical components over the first trimester and abnormal cord insertion, which contributes to the occurrence of adverse birth outcomes.
Methods: From the Japan Perinatal Registry Network database, we used data on 83 708 women who delivered singleton births at 39 cooperating hospitals in 23 Tokyo wards (2013-2015). We collected PM2.5 on a filter and measured daily concentrations of carbon and ion components. Then, we calculated the average concentrations over the first trimester (0-13 weeks of gestation) for each woman. A multilevel logistic-regression model with the hospital as a random effect was used to estimate the odds ratios (ORs) of abnormal cord insertion.
Results: Among the 83 708 women (mean age at delivery = 33.7 years), the frequency of abnormal cord insertion was 4.5%, the median concentration [interquartile range (IQR)] of total PM2.5 was 16.1 (3.61) μg/m3 and the OR per IQR for total PM2.5 was 1.14 (95% confidence interval = 1.06-1.23). In the total PM2.5-adjusted models, total carbon, organic carbon, nitrate, ammonium and chloride were positively associated with abnormal insertion. Organic carbon was consistently, and nitrate tended to be, associated with specific types of abnormal insertion (marginal or velamentous cord insertion).
Conclusions: Exposure to total PM2.5 and some of its components over the first trimester increased the likelihood of abnormal cord insertion.
Keywords: Abnormal cord insertion; fine particle; implantation; nitrate; organic carbon; pregnancy..
7. Kamura S¹'², Sasaki A¹, Ogawa K¹, Kato K², Sago H¹, Periconceptional folic acid intake and disturbing factors: A single-center study in Japan, Congenit Anom (Kyoto), 10.1111/cga.12449, 62, 1, 42-46, 2022.01, We investigated folic acid (FA) intake and disturbing factors in pregnant women who visited our center in 2017. Among 1531 pregnant women, 45.1% of women initiated FA supplementation before pregnancy. The risk of failure of supplementation was significantly lower among women of ≥35 (adjusted odds ratio [aOR] 0.43) and 30-34 years of age (aOR: 0.59) in comparison to women of <30 years of age, and among those conceived with timing/artificial insemination of husband (aOR 0.47) and in vitro fertilization/intracytoplasmic sperm injection (aOR 0.32) in comparison to those conceived naturally. The risk among those with 1 (aOR 1.44) or ≥2 previous deliveries (aOR 2.75) was significantly higher in comparison to nulliparous women. Young, multiparous women, and those with natural conception should be targeted to promote preconceptional FA intake.
Keywords: folic acid; neural tube defect (NTD); preconceptional care; risk factors; supplementation..
8. Michikawa T¹, Morokuma S², Yamazaki S³, Takami A⁴, Sugata S⁴, Yoshino A⁴, Takeda Y⁵, Nakahara K⁶, Saito S⁷, Hoshi J⁷, Kato K⁶, Nitta H³, Nishiwaki Y⁵, Exposure to chemical components of fine particulate matter and ozone, and pla-centa-mediated pregnancy complications in Tokyo: a register-based study, J Expo Sci Environ Epidemiol, 10.1038/s41370-021-00299-4, 32, 1, 135-145, 2022.01, Background: Maternal exposure to fine particulate matter (PM2.5) was associated with pregnancy complications. However, we still lack comprehensive evidence regarding which specific chemical components of PM2.5 are more harmful for maternal and foetal health.
Objective: We focused on exposure over the first trimester (0-13 weeks of gestation), which includes the early placentation period, and investigated whether PM2.5 and its components were associated with placenta-mediated pregnancy complications (combined outcome of small for gestational age, preeclampsia, placental abruption, and stillbirth).
Methods: From 2013 to 2015, we obtained information, from the Japan Perinatal Registry Network database, on 83,454 women who delivered singleton infants within 23 Tokyo wards (≈627 km2). Using daily filter sampling of PM2.5 at one monitoring location, we analysed carbon and ion components, and assigned the first trimester average of the respective pollutant concentrations to each woman.
Results: The ORs of placenta-mediated pregnancy complications were 1.14 (95% CI = 1.08-1.22) per 0.51 μg/m3 (interquartile range) increase of organic carbon and 1.11 (1.03-1.18) per 0.06 μg/m3 increase of sodium. Organic carbon was also associated with four individual complications. There was no association between ozone and outcome.
Significance: There were specific components of PM2.5 that have adverse effects on maternal and foetal health.
Keywords: Chemical component; Particulate matter; Placenta-mediated pregnancy complications; Placentation..
9. Kato K¹, Terauchi M², Annual report of the Women's Healthcare Committee, Japan Society of Obstetrics and Gynecology, 2021, J Obstet Gynaecol Res, 10.1111/jog.14986, 48, 1, 38-48, 2022.01, Our committee was founded in 2010 with the goal of improving women's health. This academic year we focused on the following five activities for the second consecutive year: cooperation between the department of pediatric surgery and the department of obstetrics and gynecology for the treatment of persistent cloaca and Mayer-Rokitansky-Küster-Hauser syndrome in Japan; assessment of the educational training of women's healthcare advisers; increasing screening for breast and cervical cancer; conducting the Nationwide Survey of Prescribing Practices for the Treatment of Menopausal Symptoms; and preventing osteoporosis in survivors of gynecologic cancer. The activities of each subcommittee are detailed below. This report is based on the Japanese version of the annual report (Acta Obst Gynaec Jpn 2021;73(6):684-699).
Keywords: Mayer-Rokitansky-Küster-Hauser syndrome; cancer screening; healthcare adviser; menopausal symptoms; osteoporosis; persistent cloaca..
10. Kikuchi K¹, Sato Y², Izukura R³, Nishikitani M⁴, Kato K⁵, Morokuma S², Nessa M⁶, Nohara Y⁷, Yokota F⁸, Ahmed A⁹, Islam-Maruf R⁴, Nakashima N⁴, Portable health clinic for sustainable care of mothers and newborns in rural Bangladesh, Comput Methods Programs Biomed, 10.1016/j.cmpb.2021.106156, 207, 106156, 2021.08, Background and objective: Regular health checkups are important for mothers and newborns to detect health problems at an early stage; however, this is often difficult in resource-limited settings. Therefore, the portable health clinic (PHC) for maternal and child health (MCH), a telemedicine health checkup system, was introduced as an intervention study in a rural area in Bangladesh. The aim of this research project was to report findings that we had observed at a mid-point of the intervention period.
Methods: This was an intervention study conducted in Shariatpur, Bangladesh. The study population included pregnant/parturient women aged 15-49 years and their newborns. With the help of the newly created PHC for MCH, health workers, with a set of sensor devices in an attaché case, visited mothers and newborns at home to examine their health status. Their health status was triaged into four categories using a data management application, and in cases of affected or emergent health status, they were placed on remote video consultation with a doctor.
Results: In total, 94 women were included in the PHC for MCH intervention. The rate of participants who received antenatal care at least four times or postnatal care at least once increased (from 29% to 51%, and from 27% to 78%, respectively) compared with before introducing PHC for MCH. Using the PHC for MCH, we detected health problems in pregnant/parturient women; a relatively high percentage had anemia (45-54%) and/or abnormal pulse rate (20-40%). Moreover, after introducing the PHC for MCH, more than 40% of women who received multiple antenatal care or postnatal care checkups improved their health status.
Conclusions: The PHC for MCH could be an effective system to improve the health of mothers and newborns by increasing the availability of care. In the future, this system is expected to be used as a primary resource for maternity healthcare, not only in rural areas but also in other social environments.
Keywords: Bangladesh; Global Health; Maternal–Child Health Services; Telemedicine..
11. Morisaki N¹, Piedvache A¹, Nagata C², Michikawa T³, Morokuma S⁴, Kato K⁵, Sanefuji M⁶,Shibata E⁷, Tsuji M⁸, Shimono M⁷, Ohga S⁹, Kusuhara K¹⁰, Japan Environment and Children’s Study Group, Maternal blood count parameters of chronic inflammation by gestational age and their associations with risk of preterm delivery in the Japan Environment and Children's Study, Sci Rep, 10.1038/s41598-021-93101-2, 11, 1, 記事番号:15522, 2021.07, Neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and lymphocyte-to-monocyte ratio (LMR), are three reportedly predictive biomarkers that reflect subclinical chronic inflammatory burden. However, how these biomarkers change during pregnancy and its clinical utility among pregnant women have been rarely studied. Among 76,853 singleton pregnancies delivered at 28-41 weeks of gestation that were enrolled in the Japan Environment and Children's Study, we observed the distribution of maternal NLR, PLR, and LMR values from week 0 to week 36 using spline curves, as well as their predictive values for preterm delivery with and without hypertensive disorders in pregnancy, placental abruption and intrauterine growth restriction (collectively termed ischemic placental disease due to their shared pathological and pathophysiological features) for measurements at 8-11 weeks, 12-17 weeks, and 18-21 weeks. NLR and PLR increased, whereas LMR decreased, with increasing gestation. High LMR and low NLR observed at 18-21 weeks, but not at earlier gestations, were associated with higher risk of preterm delivery with IPD (odds ratio 1.80 [95% CI 1.02, 3.19] per log[LMR]; odds ratio 0.49 [95% CI 0.29, 0.82] per log[NLR]). All parameters were not predictive of preterm delivery without IPD. We provide a robust reference curve for maternal blood count parameters NLR, PLR, and LMR by gestational week..
12. Yasunaga M¹, Yahata H², Okugawa K¹, Hori E¹, Kodama K¹, Yagi H¹, Ohgami T¹, Onoyama I¹, Asanoma K¹, Kato K¹, Prognostic impact of the subclassification of Müllerian cancer stage IV in the FIGO 2014 staging system with a focus of extra-abdominal lymph node metastases, Int J Clin Oncol, 10.1007/s10147-021-01908-w, 26, 7, 1330-1335, 2021.07, Background: The International Federation of Gynecology and Obstetrics (FIGO) staging system for Müllerian cancer was changed in 2014. Our objective was to evaluate the prognostic impact of stage IV subclassification in this new staging system, especially focusing on extra-abdominal lymph node metastasis.
Methods: Eighty-two patients with stage IV Müllerian cancer treated between 2005 and 2016 at our hospital were retrospectively analyzed. Data for the following clinicopathological variables were analyzed: (1) FIGO stage; (2) tumor stage; (3) lymph node status; (4) histologic type; (5) neoadjuvant chemotherapy; (6) optimal surgery; and (7) bevacizumab use. Survival analysis was performed using Kaplan-Meier curves, log-rank tests, and Cox proportional hazards models.
Results: In accordance with the new classification, 28 and 54 patients were classified as FIGO IVA and IVB, respectively. In the Cox proportional hazards model, early-stage tumors (T1b-3b) and optimal surgery were statistically significant favorable prognostic factors. However, the new FIGO system did not discriminate prognostically between stage IVA and IVB. Median overall survival of stage IVB patients diagnosed with extra-abdominal lymph node metastasis only was better than that of stage IVA and stage IVB patients diagnosed with solid organ metastasis.
Conclusions: In this analysis of the revised FIGO system of patients reclassified as FIGO stage IVA or IVB, no new prognostic information was obtained. There is a possibility that stage IVB patients diagnosed with extra-abdominal lymph node metastasis only can be classified as an earlier stage. Further modification of the FIGO staging system may be needed to improve the prediction of patient prognosis.
Keywords: Extra-abdominal lymph nodes; Müllerian cancer; Prognosis; Stage IV subclassification..
13. Nakahara K¹, Michikawa T², Morokuma S³ ⁴, Ogawa M⁵, Kato K¹ ⁵, Sanefuji M⁵ ⁶, Shibata E⁷ ⁸, Tsuji M⁷ ⁹, Shimono M⁷ ¹⁰, Kawamoto T⁷, Ohga S⁶, Kusuhara K⁷ ¹⁰, Japan Environment and Children’s Study Group, Association of maternal sleep before and during pregnancy with sleep and developmental problems in 1-year-old infants, Sci Rep, 10.1038/s41598-021-91271-7, 11, 1, 11834-11834, 2021.06, This study investigated the association of maternal sleep before and during pregnancy with sleeping and developmental problems in 1-year-old infants. We used data from the Japan Environment and Children's Study, which registered 103,062 pregnancies between 2011 and 2014. Participants were asked about their sleep habits prior to and during pregnancy. Follow-up assessments were conducted to evaluate the sleep habits and developmental progress of their children at the age of 1 year. Development during infancy was evaluated using the Ages and Stages Questionnaire (ASQ). Maternal short sleep and late bedtime before and during pregnancy increased occurrence of offspring's sleeping disturbances. For example, infants whose mothers slept for less than 6 h prior to pregnancy tended to be awake for more than 1 h (risk ratio [RR] = 1.49, 95% confidence interval [CI] 1.34-1.66), sleep less than 8 h during the night (RR = 1.60, 95% CI 1.44-1.79), and fall asleep at 22:00 or later (RR = 1.33, 95% CI 1.26-1.40). Only subjective assessments of maternal sleep quality during pregnancy, such as very deep sleep and feeling very good when waking up, were inversely associated with abnormal ASQ scores in 1-year-old infants..
14. Hachisuga K¹, Ohishi Y¹ ², Tomonobe H³, Yahata H³, Kato K³, Oda Y¹, Endometrial endometrioid carcinoma, grade 1, is more aggressive in the elderly than in the young, Histopathology, 10.1111/his.14400, 79, 5, 708-719, 2021.05, Aims: The aim of this study was to characterise grade 1 (G1) endometrioid carcinoma in the elderly, by using clinicopathological features and immunohistochemical features of surrogate markers of molecular subtypes.
Methods and results: We retrospectively analysed tumour samples from 268 patients with G1 endometrioid carcinoma (<40 years, n = 24; 40-59 years, n = 169; ≥60 years, n = 75) for whom long-term clinical follow-up data were available. G1 endometrioid carcinoma in the elderly (≥60 years) was characterised by frequent deep myometrial invasion, less frequent endometrioid intraepithelial neoplasia (EIN), lack of benign hyperplasia (BH), less frequent squamous differentiation, and occasional aberrant p53 expression. In contrast, this condition in the young (<40 years) was characterised by frequent EIN, BH, and squamous differentiation. Univariate analysis revealed that elderly status (≥60 years), International Federation of Obstetrics and Gynecology (FIGO) 2009 stage and aberrant p53 expression were significantly associated with shorter progression-free survival, and multivariate analysis revealed that elderly status and FIGO 2009 stage were independently associated with a poor prognosis.
Conclusions: G1 endometrioid carcinoma in the elderly is more aggressive than that in the young, and elderly status is an independent predictor of shorter progression-free survival in this condition. We propose that type 1 tumours can be subdivided into type 1a (young age at onset and indolent) and type 1b (old age at onset and relatively aggressive).
Keywords: G1 endometrioid carcinoma; elderly; p53 expression; shorter progression-free survival..
15. Nakahara K¹, Michikawa T², Morokuma S³⁴, Ogawa M⁵, Kato K¹⁵, Sanefuji M⁵⁶, Shibata E⁷⁸, Tsuji M⁷⁹, Shimono M⁷¹⁰, Kawamoto T⁷, Ohga S⁶, Kusuhara K⁷¹⁰, the Japan Envi-ronment and Children's Study Group, Influence of physical activity before and during pregnancy on infant's sleep and neurodevelopment at 1-year-old, Sci Rep, 10.1038/s41598-021-87612-1, 11, 1, Article number 8099, 2021.04, The aim of this study was to investigate the association between maternal physical activity (PA) before and during pregnancy and sleep and developmental problems in 1-year-old infants. We used data from a nationwide cohort study in Japan that registered 103,062 pregnancies between 2011 and 2014. Participants were asked about their PA before and during pregnancy, and the sleep and development of their children at the age of 1 year. Maternal PA was estimated using the International Physical Activity Questionnaire and was expressed in METs per week. We defined scores below the cut-off points of the Ages and Stages Questionnaire (ASQ) as abnormal for infant development. Based on the levels of PA before or during pregnancy, the participants were divided into five groups. In mothers with higher PA levels, the risk ratio for bedtime after 22:00 or abnormal ASQ scores in their 1-years-old infants were lower. These associations were observed for PA before and during pregnancy. Higher levels of maternal PA, both before and during pregnancy, may reduce sleep and developmental problems in infants..
16. Yoshida K¹, Kusama K¹, Fukushima Y¹, Ohmaru-Nakanishi T², Kato K², Tamura K¹, Alpha-1 Antitrypsin-Induced Endoplasmic Reticulum Stress Promotes Invasion by Extravillous Trophoblasts, INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 10.3390/ijms22073683, 22, 7, 記事番号:3683, 2021.04, Alpha-1 antitrypsin (A1AT) is a glycoprotein that has been shown to protect tissues from proteolytic damage under various inflammatory conditions. Several studies show that A1AT may be associated with pre-eclampsia. However, the role of A1AT expression in placental physiology is not fully understood. In the present study, we aim to characterize the expression and function of placental A1AT. A1AT knockdown is found to reduce the expression of the serine protease HTRA1 in a trophoblast cell line. In addition, A1AT overexpression (A1AT-OE) increases the expression of HTRA1, IL6, CXCL8, and several markers of endoplasmic reticulum (ER) stress. Treatment with tunicamycin or thapsigargin, which induces ER stress, increases HTRA1 expression. Furthermore, immunohistochemistry reveals that HTRA1 is expressed in trophoblasts and the endometrial decidual cells of human placentas. An invasion assay shows that A1AT and HTRA1 stimulate cell invasion, but treatment with the ER stress inhibitors reduces the expression of HTRA1 and ER stress markers and prevents cell invasion in A1AT-OE trophoblasts. These results suggest that endogenous A1AT regulates inflammatory cytokine expression and HTRA1-induced trophoblast invasion via the induction of ER stress. It is concluded that an imbalance in the functional link between A1AT and ER stress at the maternal-fetal interface might cause abnormal placental development.
Keywords: alpha-1 antitrypsin; cell invasion; endoplasmic reticulum stress; extravillous trophoblast; high-temperature requirement A serine peptidase 1..
17. Sato Y¹, Hidaka N², Sakai A³, Kido S⁴, Fujita Y⁵, Okugawa K⁶, Yahata H⁷, Kato K⁸, Evaluation of the efficacy of vaginal progesterone in preventing preterm birth after abdominal trachelectomy, Eur J Obstet Gynecol Reprod Biol, 10.1016/j.ejogrb.2021.02.009, 259, 119-124, 2021.04, Objective: To determine whether vaginal progesterone (VP) reduces the rate of preterm birth in pregnant women after abdominal trachelectomy (AT) for early-stage cervical cancer STUDY DESIGN: This is an interventional study with a historical cohort. For the interventional study participants who had singleton pregnancies after AT between October 2016 and September 2020, the administration of vaginal progesterone was started between 16+ and 19+6 weeks of gestation and discontinued at 34 weeks of gestation or at the time of delivery, rupture of membranes, or massive uterine bleeding. We investigated obstetric and neonatal outcomes among the study participants and compared them with outcomes of the historical control group participants, included women with singleton pregnancies after AT who were managed without VP at our institution between January 2007 and September 2016, using Fisher's exact test and the Mann-Whitney U test The main outcomes were the gestational age at delivery and incidence of preterm birth before 37 weeks and 34 weeks of gestation.
Result: Twelve pregnancies in ten women were included in the VP group. In contrast, 19 pregnancies in 17 women were included in the historical control group. The incidence of preterm birth at <37 weeks was 10/12 (83 %) in the VP group and 11/19 (58 %) in the control group. The incidence of preterm birth at <34 weeks was 6/12 (50 %) in the VP group and 9/19 (48 %) in the control group. The incidence of preterm birth in the two groups was similar, and the difference between the two groups was not statistically significant.
Conclusion: The administration of vaginal progesterone did not reduce the rate of preterm birth among pregnant women after AT.
Keywords: Abdominal trachelectomy; Preterm birth; Preterm premature membranes rupture; Vaginal progesterone..
18. Sonoda K¹ ², Ohgami T², Hachisuga M², Fujita Y², Okugawa K², Yahata H², Kato K², Difficulty of cervical cancer diagnosis during pregnancy: A case series analysis of the clinicopathological characteristics and prognosis of cervical cancer diagnosed during pregnancy or within 6 months after parturition, Mol Clin Oncol, 10.3892/mco.2021.2229, 14, 4, Article number 67, Pages 1-7, 2021.04, Due to the recent trend of women having children at an older age, the number of pregnancies complicated by cervical cancer has increased. In the present study, the clinical characteristics of patients with cervical cancer complicated by pregnancy were analyzed. The clinicopathological characteristics and prognosis of patients with cervical cancer during pregnancy who underwent treatment at Kyushu University Hospital from January 2008 to December 2017 were assessed retrospectively from their medical records. The medical information of patients diagnosed within 6 months after parturition was also evaluated as these patients were considered to be affected by cervical cancer during pregnancy. A total of 19 women were diagnosed with cervical cancer during pregnancy (median age, 33 years), three of whom were diagnosed as negative for intraepithelial lesion or malignancy at the initial visit to a previous clinic after pregnancy. The tumor stage was IA1 in one patient, IB1 in 16 patients, IB2 in one patient and IVB in one patient. The median gestational age at the time of cervical cancer diagnosis was 13 weeks. One patient died of cervical cancer during the follow-up period. An additional 12 patients were diagnosed within 6 months after parturition. The median age of these patients was 35.5 years. Cancer screening was performed in 10 patients during pregnancy, none of whom were diagnosed with cervical cancer, including three patients who were negative for intraepithelial lesion or malignancy. Of the 12 patients, one had tumor stage IA1, eight had IB1, two had IB2 and one had IIB. Three patients experienced recurrence, of whom one died of cervical cancer. Advanced cervical cancer was diagnosed both during pregnancy and within 6 months after parturition. Diagnosis of cervical cancer during pregnancy is difficult even at an advanced stage; however, rapid diagnosis and prompt multidisciplinary treatment are critical. Therefore, it is necessary to improve the accuracy of cervical cancer diagnosis, and to characterize tumor cells and their microenvironment, during pregnancy.
Keywords: diagnosis; parturition; pregnancy; prognosis; uterine cervical neoplasms..
19. Kodama K¹, Yahata H², Okugawa K¹, Tomonobe H¹, Yasutake N¹, Yoshida S¹, Yagi H¹, Yasunaga M¹, Ohgami T¹, Onoyama I¹, Asanoma K¹, Hori E¹, Shimokawa M³, Kato K¹, Prognostic outcomes and risk factors for recurrence after laser vaporization for cervical intraepithelial neoplasia: a single-center retrospective study, Int J Clin Oncol, 10.1007/s10147-020-01848-x, 26, 4, 770-776, 2021.04, Background: Cervical intraepithelial neoplasia (CIN) is a precancerous lesion that may progress to invasive cervical cancer without intervention. We aim to examine the prognostic outcomes and risk factors for recurrence after laser vaporization for CIN 3, CIN 2 with high-risk human papillomavirus (HPV) infection, and CIN 1 persisting for more than 2 years.
Methods: Between 2008 and 2016, a total of 1070 patients underwent cervical laser vaporization using a carbon dioxide laser. We performed a retrospective review of their medical records to assess their clinical characteristics, pathologic factors, and prognostic outcomes.
Results: The mean patient age was 34 years (range 18-64 years). The preoperative diagnosis was CIN 1 in 27 patients, CIN 2 in 485 patients, and CIN 3 in 558 patients. Over a median follow-up period of 15 months, the 2-year recurrence rate was 18.9%, and the 5-year recurrence rate was 46.5%. The 2-year retreatment rate was 12.6%, and the 5-year retreatment rate was 30.5%. We diagnosed 9 patients with invasive cancer after treatment; all patients underwent combined multidisciplinary treatment, and there were no deaths during follow-up. The recurrence-free interval was correlated with patient age (hazard ratio [HR], 1.028; 95% CI 1.005-1.051; P = 0.0167), body mass index (HR, 1.052; 95% CI 1.008-1.098; P = 0.0191), and glandular involvement (HR, 1.962; 95% CI 1.353-2.846; P = 0.0004).
Conclusions: Cervical laser vaporization is effective and useful for patients with CIN who wish to preserve fertility. However, patients with glandular involvement, older age, and higher body weight require close follow-up for recurrence.
Keywords: Cervical intraepithelial neoplasia; Laser vaporization; Recurrence-free interval..
20. Yamaguchi S¹, Yahata H¹, Okugawa K¹, Kodama K¹, Yagi H¹, Yasunaga M¹, Ohgami T¹, Onoyama I¹, Asanoma K¹, Kato K¹, Hypersensitivity reaction to pegylated liposomal doxorubicin administration for Mullerian carcinoma in Japanese women, J Obstet Gynaecol Res, 10.1111/jog.14680, 47, 4, 1544-1548, 2021.04, Aim: PEGylated liposomal doxorubicin (PLD) is a therapeutic agent for gynecological malignancy. Hypersensitivity reaction (HSR) is a major adverse effect that usually disappears after halting administration of PLD. Premedication is usually not necessary before administration of PLD to prevent HSR. Here, we evaluated the frequency of HSR during administration of PLD following premedication in Japanese women.
Methods: We performed PLD administration in 78 patients (386 cycles) between 2013 and 2018. Granisetron hydrochloride and dexamethasone sodium phosphate were administered 30 min before PLD administration. Then, PLD (40 or 30 mg/m2 combined usage with carboplatin) was administered. We retrospectively reviewed the medical records of 78 patients and examined the frequency of HSR.
Results: Seven of 78 (9%) patients showed HSR by PLD administration following premedication. One patient showed cardiopulmonary arrest in 13 min after PLD administration (grade 4). The other six patients showed grade 2 HSR. All patients developed HSR in the first course. The incidence of HSR was significantly higher in patients with allergic history than in patients without allergic history (p = 0.0151).
Conclusions: Clinicians should be aware of the potential for HSR in patients administered PLD, particularly those with allergic history and those receiving the first cycle of PLD, even following premedication..
21. Takamatsu K¹, Ogawa M¹, Obayashi S², Takeda T³, Terauchi M⁴, Higuchi T⁵, Kato K⁶, Kubota T⁷, A Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial to Investigate the Effects of Kamishoyosan, a Traditional Japanese Medicine, on Menopausal Symptoms: The KOSMOS Study, Evid Based Complement Alternat Med ., 10.1155/2021/8856149, 2021, 8856149-8856149, 2021.02, Objective: The KOSMOS study, a multicenter, randomized, double-blind, placebo-controlled trial, investigated the effects and safety of kamishoyosan (TJ-24), a traditional Japanese medicine, in the treatment of climacteric disorder.
Methods: Japanese women with climacteric disorder were administered a placebo during a 4-week run-in period, after which they were classified as placebo responders (R group) if their score on the modified Questionnaire for the Assessment of Climacteric Symptoms in Japanese Women (m-QACS) with excitability and irritability as the primary outcome improved by ≥ 3 points and as placebo nonresponders (NR group) otherwise. Members of the NR group were randomly allocated to receive either TJ-24 or placebo. After 12 weeks, their m-QACS scores, anxiety and depression, sleep, and overall quality of life (QOL) were compared.
Results: The TJ-24 and placebo arms in the NR group included 20 patients each. The change in the m-QACS scores of members of the NR group for excitability and irritability at 12 weeks versus baseline was -3.1 ± 1.7 in the TJ-24 arm, a significant decrease, but compared with -2.7 ± 2.2 in the placebo arm, no significant difference was between two arms. However, the proportion of participants whose score improved by ≥3 points was significantly higher in the TJ-24 arm. In the subgroup analysis of premenopausal women, the changes in the score for excitability and irritability were significantly larger in the TJ-24 arm. The incidence of adverse drug reactions or adverse events did not differ between the two arms, and no serious events were reported.
Conclusion: Although no significant difference was identified for the primary outcome, a significantly higher proportion of patients who received TJ-24 displayed improvement. Its high level of safety and effects on excitability and irritability in premenopausal women suggest that TJ-24 may be a useful treatment..
22. Suzuki I¹ ²,Yoshida S², Tabu K³, Kusunoki¹S¹, Matsumura Y², Izumi H⁴, Asanoma K², Yagi H², Onoyama I², Sonoda K⁵, Kohno K⁶, Taga T³, Itakura A¹, Takeda S¹, Kato K⁷, YBX2 and cancer testis antigen 45 contribute to stemness, chemoresistance and a high degree of malignancy in human endometrial cancer, Sci Rep, 10.1038/s41598-021-83200-5, 11, 1, 4220-4220, 2021.02, Y-box binding protein 2 (YBX2) has been associated with the properties of both germ cells and cancer cells. We hypothesized that YBX2 might contribute to the characteristics of cancer stem cells (CSCs). In this study, we clarified the function of YBX2 in endometrial cancer stem cells. We established a human YBX2-expressing Ishikawa (IK) cell line (IK-YBX2 cells). We analyzed gene expression associated with stemness and isolated SP cells from IK-YBX2 cells. The SP population of IK-YBX2 cells, the expression of ALDH1 and serial sphere-forming capacity were associated with levels of YBX2 expression. IK-YBX2 cells were resistant to anti-cancer drugs. In gene expression analysis, a gene for cancer testis antigen, CT45, was generally overexpressed in IK-YBX2 cells. YBX2-mediated CT45 expression was associated with increased levels of self-renewal capacity and paclitaxel resistance. The level of CT45 expression was enhanced in high-grade and/or advanced stages of human endometrial cancer tissues. We conclude that expression of YBX2 is essential for the stem cell-like phenotype. CT45 contributes to stemness associated with YBX2 and might be related to the progression of endometrial cancer..
23. Kawamura T¹, Tomari H¹, Onoyama I¹, Araki H², Yasunaga M¹, Lin C¹, Kawamura K¹, Yokota N¹, Yoshida S¹, Yagi H¹, Asanoma K¹, Sonoda K¹, Egashira K¹, Ito T², Kato K¹, Identification of genes associated with endometrial cell ageing, Mol Hum Reprod, 10.1093/molehr/gaaa078, 27, 2, gaaa078-gaaa078, 2021.02, Ageing of the uterine endometrium is a critical factor that affects reproductive success, but the mechanisms associated with uterine ageing are unclear. In this study, we conducted a qualitative examination of age-related changes in endometrial tissues and identified candidate genes as markers for uterine ageing. Gene expression patterns were assessed by two RNA-sequencing experiments using uterine tissues from wild type (WT) C57BL/6 mice. Gene expression data obtained by RNA-sequencing were validated by real-time PCR. Genes expressing the pro-inflammatory cytokines Il17rb and chemokines Cxcl12 and Cxcl14 showed differential expression between aged WT mice and a group of mice composed of 5- and 8-week-old WT (young) animals. Protein expression levels of the above-mentioned genes and of IL8, which functions downstream of IL17RB, were analysed by quantitative immunohistochemistry of unaffected human endometrium tissue samples from patients in their 20s and 40s (10 cases each). In the secretory phase samples, 3,3'- diaminobenzidine staining intensities of IL17RB, CXCL12 and CXCL14 for patients in their 40s were significantly higher than that for patients in their 20s, as detected by a Mann-hitney U test. These results suggest that these genes are candidate markers for endometrial ageing and for prediction of age-related infertility, although confirmation of these findings is needed in larger studies involving fertile and infertile women.
Keywords: CXCL12; CXCL14; IL17RB; endometrial cell ageing; infertility; quantitative immunohistochemistry..
24. Yoshida S¹, Asanoma K¹, Yagi H¹, Onoyama I¹, Hori E¹, Matsumura Y¹, Okugawa K¹, Yahata H¹, Kato K¹, Fibronectin mediates activation of stromal fibroblasts by SPARC in endometrial cancer cells, BMC Cancer, 10.1186/s12885-021-07875-9, 21, 1, 156-156, 2021.02, Background: Matricellular glycoprotein, SPARC is a secreted molecule, that mediates the interaction between cells and extracellular matrix. SPARC functions as a regulator of matrix organization and modulates cell behavior. In various kinds of cancer, strong SPARC expression was observed in stromal tissues as well as in cancer epithelial cells. The function of SPARC in cancer cells is somewhat controversial and its impact on peritumoral stromal cells remains to be resolved.
Methods: We investigated the effects of SPARC expression in endometrial cancer cells on the surrounding stromal fibroblasts using in vitro co-culture system. Changes in characteristics of fibroblasts were examined by analysis of fibroblast-specific markers and in vitro contraction assay.
Results: SPARC induced AKT phosphorylation and epithelial-to-mesenchymal transition, consistent with previous reports. Cancer-associated fibroblasts of endometrial cancer expressed higher levels of mesenchymal- and fibroblast-associated factors and had a stronger contraction ability. Unexpectedly, cancer-associated fibroblasts expressed comparable levels of SPARC compared with fibroblasts from normal endometrium. However, co-culture of normal fibroblasts with SPARC-expressing Ishikawa cells resulted in activation of the fibroblasts. Immunodepletion of SPARC did not affect the activation of fibroblasts.
Conclusions: Our data indicated that SPARC activated fibroblasts only in the presence of fibronectin, which was abundantly secreted from SPARC-expressing endometrial cancer cells. These results suggested that a SPARC-fibronectin-mediated activation of fibroblasts might be involved in enhanced mobility and invasion of cancer cells..
25. Okugawa K¹, Yahata H², Sonoda K³, Kodama K², Yagi H², Ohgami T², Yasunaga M², Onoyama I², Kaneki E², Asanoma K², Kobayashi H⁴, Kato K², Evaluation of adjuvant chemotherapy after abdominal trachelectomy for cervical cancer: a single-institution experience, Int J Clin Oncol, 10.1007/s10147-020-01778-8, 26, 1, 216-224, 2021.01, Background: To avoid the loss of fertility, chemotherapy should be chosen as an adjuvant treatment after trachelectomy. Our study evaluated the effectiveness and safety of adjuvant chemotherapy after abdominal trachelectomy for cervical cancer.

Methods: Our institutional review board approved this clinical study, and informed consent was obtained from each patient. We began performing abdominal trachelectomy at our institution in 2005. Deep stromal invasion (more than two-thirds) with lymphovascular space invasion, diffuse cervical invasion, skip lesions in the vagina, and lymphovascular space invasion in the cardinal ligament and vagina were defined as intermediate-risk factors, and parametrial invasion and pelvic lymph node metastasis were defined as high-risk factors. Patients who had intermediate- or high-risk factors received post-trachelectomy adjuvant treatment. The medical records and information of the patients were reviewed retrospectively.

Results: Through January 2020, we performed 212 trachelectomies. Among the included patients, 16 and 7 patients with intermediate- and high-risk cancer, respectively, received adjuvant chemotherapy after trachelectomy (2 and 21 patients underwent abdominal modified radical trachelectomy and radical trachelectomy, respectively). Among these patients, only one (4.3%) experienced relapse and subsequent death of the disease after a median postoperative follow-up of 80 months (range 12-146 months). The 5-year survival rate was 95.5%. Chemotherapy-related life-threatening acute adverse events were not observed. Persistent ovarian dysfunction and late adverse events did not occur. One woman achieved three pregnancies, and two infants were delivered.

Conclusion: Adjuvant chemotherapy after abdominal trachelectomy could be an alternative treatment option from the aspects of effectiveness, safety, and fertility preservation..
26. Kato K¹, Terauchi M², Annual report of the Women's Health Care Committee, Japan Society of Obstet-rics and Gynecology, 2020, J Obstet Gynaecol Res, 10.1111/jog.14417, 47, 1, 52-62, 2021.01, Our committee was founded in 2010 with the goal of improving women's health. This academic year, we focused on five activities: cooperation between the departments of pediatric surgery and obstetrics and gynecology for the treatment of persistent cloaca and Mayer-Rokitansky-Küster-Hauser syndrome in Japan; assessment of the educational training of women's health-care advisers; increasing screening for breast and cervical cancer; conducting the Nationwide Survey of Prescribing Practices for the Treatment of Menopausal Symptoms; and preventing osteoporosis in survivors of gynecologic cancer. The activities of each subcommittee are detailed below. This report is based on the Japanese version of the annual report (Acta Obst Gynaec Jpn 2020;72(6):697-707)..
27. Yahata H, Sonoda K, Inoue S, Yasutake N, Kodama K, Yagi H, Yasunaga M, Ohgami T, Onoyama I, Kaneki E, Okugawa K, Asanoma K, Kato K, Is Adjuvant Therapy Necessary for Patients with Intermediate-Risk Cervical Cancer after Open Radical Hysterectomy?, Oncology, 10.1159/000508569, 98, 12, 853-858, 2020.11, Introduction: Adjuvant therapy is usually recommended for patients with intermediate-risk cervical cancer (deep stromal invasion [DSI], lymphovascular space invasion [LVSI], and bulky tumor) after radical hysterectomy. However, we previously reported that DSI, LVSI, and bulky squamous cell carcinoma (SCC) were not correlated with prognosis in multivariate analysis; therefore, the indications we use for adjuvant therapy include complete stromal invasion, not DSI or LVSI or bulky SCC. The objective of this study was to evaluate the adequacy of our therapeutic strategy for cervical cancer after radical hysterectomy.
Methods: We performed 321 type III open radical hysterectomies for cervical cancer between 2001 and 2013. Eighty-two patients with DSI, LVSI, or bulky SCC did not receive adjuvant therapy after radical hysterectomy under informed consent. We retrospectively evaluated the prognosis of these 82 patients.
Results: Forty-two patients had >2/3 DSI and 35 patients had 1/3-2/3 DSI. Five patients had LVSI alone. The mean patient age was 43 years (range, 27-72). Six patients (7%) experienced recurrence during a median follow-up period of 84 months (range, 1-206). Two of the 6 patients with recurrence suffered cervical cancer-related deaths, but the remaining 4 cases are alive without evidence of disease after treatment during a follow-up period of 87-165 months. The 5-year disease-free survival rate was 92.6%, and the 5-year overall survival rate was 96.3%.
Conclusions: Adjuvant therapy for DSI, LVSI, or bulky SCC after open radical hysterectomy might not be necessary. Further data collection is warranted to determine the standard of care for patients with intermediate-risk cervical -cancer..
28. Kato M¹ ², Onoyama I¹, Kawakami M¹, Yoshida S¹, Kawamura K¹, Kodama K¹, Hori E¹, Cui L¹, Matsumura Y¹, Yagi H¹, Asanoma K¹, Yahata H¹, Itakura A², Takeda S², Kato K¹, Downregulation of 5-hydroxymethylcytosine is associated with the progression of cervical intraepithelial neoplasia, PLoS One, 10.1371/journal.pone.0241482, 15, 11, e0241482-e0241482, 2020.10, Around the world, cervical cancer is one of the most common neoplastic diseases among women, and the prognosis of patients in an advanced stage remains poor. To reduce the mortality rate of cervical cancer, early diagnosis and treatment are essential. DNA methylation is an important aspect of gene regulation, and aberrant DNA methylation contributes to carcinogenesis and cancer progression in various cancers. Although 5-methylcytosine (5mC) has been analyzed intensively, the function of 5-hydroxymethylcytosine (5hmC) has not been clarified. The purpose of our study was to identify the molecular biomarkers for early diagnosis of cervical tumors due to epigenetic alterations. To assess the clinical relevance of DNA methylation, we used immunohistochemistry (IHC) to characterize the level of 5hmC in 102 archived human cervical intraepithelial neoplasia (CIN) samples and cervical cancer specimens. The level of 5hmC was significantly decreased between CIN2 and CIN3. The progression of cervical tumors is caused by a reduction of TP53 and RB1 because of HPV infection. We observed that Tp53 and Rb1 were knocked down in mouse embryonic fibroblasts (MEF), a model of normal cells. The level of 5hmC was reduced in Tp53-knockdown cells, and the expression levels of DNA methyltransferase 1 (DNMT1) and ten-eleven translocation methylcytosine dioxygenase 1 (TET1) were induced. In contrast, there was no significant change in Rb1-knockdown cells. Mechanistically, we focused on apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like (APOBEC) 3B (A3B) as a cause of 5hmC reduction after TP53 knockdown. In the human cell line HHUA with a wild-type TP53 gene, A3B was induced in TP53-knockdown cells, and A3B knockdown recovered 5hmC levels in TP53-knockdown cells. These data indicate that TP53 suppression leads to 5hmC reduction in part through A3B induction. Moreover, IHC showed that expression levels of A3B in CIN3 were significantly higher than those in both normal epithelium and in CIN2. In conclusion, 5hmC levels are decreased between CIN2 and CIN3 through the TP53-A3B pathway. Since A3B could impair genome stability, 5hmC loss might increase the chances of accumulating mutations and of progressing from CIN3 to cervical cancer. Thus, these epigenetic changes could predict whether CINs are progressing to cancer or disappearing..
29. Kato M¹ ², Onoyama I¹, Yoshida S¹, Cui L¹, Kawamura K¹, Kodama K¹, Hori E¹, Matsumura Y¹, Yagi H¹, Asanoma K¹, Yahata H¹, Itakura A², Takeda S², Kato K¹, Dual-specificity Phosphatase 6 Plays a Critical Role in the Maintenance of a Can-cer Stem-Like Cell Phenotype in Human Endometrial Cancer, Int J Cancer, 10.1002/ijc.32965, 147, 7, 1987-1999, 2020.10, The prognosis of patients with high-grade or advanced-stage endometrial cancer remains poor. As cancer stem-like cells (CSCs) are thought to be associated with endometrial cancers, it is essential to investigate the molecular mechanisms that regulate endometrial CSCs. Dual-specificity phosphatase 6 (DUSP6) functions as a negative-feedback regulator of MAPK-ERK1/2 signaling, but its role in endometrial cancer remains unknown. We investigated whether DUSP6 is involved in cancer cell stemness using endometrial cancer cell lines and specimens from endometrial cancer patients. DUSP6 induced the expression of CSC-related genes including ALDH1, Nanog, SOX2 and Oct4A, increased the population of cells in the G0/G1 phase, and promoted sphere formation ability. DUSP6 knockdown resulted in reduced cell invasion and metastasis, whereas DUSP6 overexpression inhibited apoptosis under serum-free conditions. Moreover, DUSP6 decreased phosphorylated ERK1/2 and increased phosphorylated Akt levels, which potentially induces CSC features. In patients with endometrial cancers, DUSP6 expression was determined using immunohistochemistry, and based on the results, the patients were dichotomized into high- and low-DUSP6-expression groups. Progression-free survival and overall survival were significantly shorter in the high-DUSP6-expression group. These results suggest that DUSP6 has potential value as a biomarker of CSCs and as a target of therapies designed to eliminate CSCs in endometrial cancer..
30. Matsui T¹, Tamoto R², Iwasa A², Mimura M², Taniguchi S¹, Hasegawa T¹, Sudo T¹, Mizuno H¹, Kikuta J¹, Onoyama I³, Okugawa K³, Shiomi M⁴, Matsuzaki S⁴, Morii E⁵, Kimura T⁴, Kato K³, Kiyota Y², Ishii M⁶, Nonlinear Optics with Near-Infrared Excitation Enable Real-Time Quantitative Diagnosis of Human Cervical Cancers, Cancer Res, 10.1158/0008-5472.CAN-20-0348, 80, 17, 3745-3754, 2020.09, Histopathologic analysis through biopsy has been one of the most useful methods for the assessment of malignant neoplasms. However, some aspects of the analysis such as invasiveness, evaluation range, and turnaround time from biopsy to report could be improved. Here, we report a novel method for visualizing human cervical tissue three-dimensionally, without biopsy, fixation, or staining, and with sufficient quality for histologic diagnosis. Near-infrared excitation and nonlinear optics were employed to visualize unstained human epithelial tissues of the cervix uteri by constructing images with third-harmonic generation (THG) and second-harmonic generation (SHG). THG images enabled evaluation of nuclear morphology in a quantitative manner with six parameters after image analysis using deep learning. It was also possible to quantitatively assess intraepithelial fibrotic changes based on SHG images and another deep learning analysis. Using each analytical procedure alone, normal and cancerous tissue were classified quantitatively with an AUC ≥0.92. Moreover, a combinatory analysis of THG and SHG images with a machine learning algorithm allowed accurate classification of three-dimensional image files of normal tissue, intraepithelial neoplasia, and invasive carcinoma with a weighted kappa coefficient of 0.86. Our method enables real-time noninvasive diagnosis of cervical lesions, thus constituting a potential tool to dramatically change early detection. SIGNIFICANCE: This study proposes a novel method for diagnosing cancer using nonlinear optics, which enables visualization of histologic features of living tissues without the need for any biopsy or staining dye..
31. Nakahara K¹, Michikawa T², Morokuma S³ ⁴, Ogawa M⁵, Kato K¹ ⁵, Sanefuji M⁵ ⁶, Shibata E⁷ ⁸, Tsuji M⁷ ⁹, Shimono M⁷ ¹⁰, Kawamoto T⁷, Ohga S⁶, Kusuhara K⁷ ¹⁰, Japan Environment and Children’s Study Group, Non-reassuring foetal status and sleep problems in 1-year-old infants in the Japan Environment and Children's Study: a cohort study, Sci Rep, 10.1038/s41598-020-67856-z, 10, 1, 11432-11432, 2020.07, Abnormal autonomic function may cause false-positive non-reassuring foetal status (fpNRFS) and may also cause sleeping problems after birth. However, an association between fpNRFS and sleeping problems in infants has not been reported. We previously showed an association of NRFS with temperament, including bad mood and frequent crying for long durations in 1-month-old infants. In the present study, we aimed to assess this association in 1-year-old infants. A total of 62,612 single pregnant women were included in the analysis. fpNRFS was identified from medical records. Sleep problems, such as short sleep duration or crying at night, were investigated in 1-year-old infants using a questionnaire for mothers. We used a log-binominal regression model to explore the association of fpNRFS with each sleep problem and to estimate risk ratios (RRs). The number of fpNRFS cases was 2,071, with a frequency of 3.3%. We observed an association of fpNRFS with shorter sleep duration of less than 8 h a night (RR 1.30, 95% confidence intervals [CI] 1.10-1.54), crying at night (RR 1.19, 95% CI 1.03-1.39), and bedtime after 22:00 (RR 1.09, 95% CI 1.00-1.18). fpNRFS may be associated with sleep problems in 1-year-old infants..
32. Nakahara K¹, Michikawa T², Morokuma S³ ⁴, Ogawa M⁵, Kato K¹ ⁵, Sanefuji M⁵ ⁶, Shibata E⁷ ⁸, Tsuji M⁷ ⁹, Shimono M⁷ ¹⁰, Kawamoto T⁷, Ohga S⁶, Kusuhara K⁷ ¹⁰, Japan Environment and Children’s Study Group, Association of maternal sleep before and during pregnancy with preterm birth and early infant sleep and temperament, Sci Rep, 10.1038/s41598-020-67852-3, 10, 1, 11084-11084, 2020.07, This study aimed to investigate the association of maternal sleep before and during pregnancy with preterm birth, infant sleep and temperament at 1 month of age. We used the data of the Japan Environment and Children's Study, a cohort study in Japan, which registered 103,099 pregnancies between 2011 and 2014. Participants were asked about their sleep before and during pregnancy, and the sleep and temperament of their newborns at 1 month of age. Preterm birth data were collected from medical records. Maternal sleep was not associated with preterm birth, but subjective sleep quality during pregnancy was associated with late preterm birth (birth at 34-36 weeks of gestation). For example, participants with extremely light subjective depth of sleep were more likely to experience preterm birth (RR = 1.19; 95% confidence interval [CI] = 1.04-1.35). Maternal sleep both before and during pregnancy seemed to be associated with infant sleep and temperament at 1 month of age. Infants, whose mothers slept for less than 6 hours before pregnancy, tended to cry intensely (RR = 1.15; 95% CI = 1.09-1.20). Maternal sleep problems before and during pregnancy were associated with preterm birth and child sleep problems and temperament..
33. Okugawa K¹, Yahata H¹, Sonoda K¹, Ohgami T¹, Yasunaga M¹, Kaneki E¹, Kato K¹, Safety evaluation of abdominal trachelectomy in patients with cervical tumors ≥2 cm: a single-institution, retrospective analysis, J Gynecol Oncol, 10.3802/jgo.2020.31.e41, 31, 4, e41-e41, 2020.07, Objective: For oncologic safety, vaginal radical trachelectomy is generally performed only in patients with cervical cancers smaller than 2 cm. However, because inclusion criteria for abdominal trachelectomy are controversial, we evaluated the safety of abdominal trachelectomy for cervical cancers ≥2 cm.
Methods: We began performing abdominal trachelectomies at our institution in 2005, primarily for squamous cell carcinoma ≤3 cm or adenocarcinoma/adenosquamous carcinoma ≤2 cm. If a positive sentinel lymph node or cervical margin was diagnosed intraoperatively by frozen section, the trachelectomy was converted to a hysterectomy. Medical records of these patients were reviewed retrospectively. Patients who had undergone simple abdominal trachelectomy were excluded from this study.
Results: We attempted trachelectomy in 212 patients. Among the 135 patients with tumors <2 cm, trachelectomy was successful in 120, one of whom developed recurrence and none of whom died of their disease. Among 77 patients with tumors ≥2 cm, trachelectomy was successful in 62, 2 of whom developed recurrence and 1 of whom died of her disease. The overall relapse rate after trachelectomy was 1.6% (0.8% in <2 cm group and 3.2% in ≥2 cm group), and the mortality rate was 0.5% (0% in <2 cm group and 1.6% in ≥2 cm group). Recurrence-free survival (p=0.303) and overall survival (p=0.193) did not differ significantly between the <2 cm and ≥2 cm groups.
Conclusions: Abdominal trachelectomy with intraoperative frozen sections of sentinel lymph nodes and cervical margins is oncologically safe, even in patients with tumors ≥2 cm..
34. Tomari H¹ ², Kawamura T¹, Asanoma K¹, Egashira K¹, Kawamura K¹, Honjo K², Nagata Y², Kato K¹, Contribution of Senescence in Human Endometrial Stromal Cells During Prolifer-ative Phase to Embryo Receptivity, Biol Reprod, 10.1093/biolre/ioaa044, 103, 1, 104-113, 2020.06, Successful assisted reproductive technology pregnancy depends on the viability of embryos and endometrial receptivity. However, the literature has neglected effects of the endometrial environment during the proliferative phase on implantation success or failure. Human endometrial stromal cells (hESCs) were isolated from endometrial tissues sampled at oocyte retrieval during the proliferative phase from women undergoing infertility treatment. Primary hESC cultures were used to investigate the relationship between stemness and senescence induction in this population and embryo receptivity. Patients were classified as receptive or non-receptive based on their pregnancy diagnosis after embryo transfer. Biomarkers of cellular senescence and somatic stem cells were compared between each sample. hESCs from non-receptive patients exhibited significantly higher (p < 0.01) proportions of senescent cells, mRNA expressions of CDKN2A and CDKN1A transcripts (p < 0.01), and expressions of genes encoding the senescence-associated secretory phenotype (p < 0.05). hESCs from receptive patients had significantly higher (p < 0.01) mRNA expressions of ABCG2 and ALDH1A1 transcripts. Our findings suggest that stemness is inversely associated with senescence induction in hESCs, and by extension, that implantation failure in infertility treatment may be attributable to a combination of senescence promotion and disruption of this maintenance function in this population during the proliferative phase of the menstrual cycle. This is a promising step towards potentially improving the embryo receptivity of endometrium. The specific mechanism by which implantation failure is prefigured by a loss of stemness among endometrial stem cells, and cellular senescence induction among hESCs, should be elucidated in detail in the future..
35. Nitahara K¹, Hidaka N¹, Sakai A¹, Kido S¹, Kato K¹, The impact of general anesthesia on mother-infant bonding for puerperants who undergo emergency cesarean deliveries, J Perinat Med, 10.1515/jpm-2019-0412, 48, 5, 463-470, 2020.06, Background Mother-infant bonding is an emerging perinatal issue. While emergency cesarean deliveries are associated with a risk of bonding disorders, the mode of anesthesia used for emergency cesarean deliveries has never been studied in this context. We aimed to investigate the impact of administering general anesthesia and neuraxial anesthesia to women undergoing cesarean deliveries on mother-infant bonding. Methods This was a retrospective, propensity score-matched multivariable analysis of 457 patients who underwent emergency cesarean deliveries between February 2016 and January 2019 at a single teaching hospital in Japan. The Mother-Infant Bonding Scale (MIBS) scores at hospital discharge and the 1-month postpartum outpatient visit were evaluated in the general anesthesia and the neuraxial anesthesia groups. A high score on the MIBS indicates impaired mother-infant bonding. Results The primary outcome was the MIBS score at hospital discharge in propensity score-matched women. After propensity score matching, the median [interquartile range (IQR)] MIBS scores were significantly higher in the general anesthesia group than those in the neuraxial anesthesia group at hospital discharge [2 (1-4) vs. 2 (0-2); P = 0.015] and at the 1-month postpartum outpatient visit [1 (1-3) vs. 1 (0-2); P = 0.046]. In linear regression analysis of matched populations, general anesthesia showed a significant and positive association with the MIBS scores at hospital discharge [beta coefficient 0.867 (95% confidence interval [CI] 0.147-1.59); P = 0.019] but not at the 1-month postpartum outpatient visit [0.455 (-0.134 to 1.044); P = 0.129]. Conclusion General anesthesia for emergency cesarean delivery is an independent risk factor associated with impaired mother-infant bonding..
36. Maehara K¹, Morokuma S², Nakahara K¹, Okawa H¹, Kato K¹, A study on the association between eye movements and regular mouthing movements (RMMs) in normal fetuses between 24 to 39 weeks of gestation, PLoS One, 10.1371/journal.pone.0233909, 15, 5, e0233909-e0233909, 2020.05, Regular Mouthing Movements (RMMs) are movements in which lips and lower jaw movements occur regularly and can be observed in the fetus using transabdominal ultrasonic tomography. In near term infants, it is known that RMMs form clusters during the quiet sleep period. The notation of RMMs is not uniform, and is described as spontaneous sucking movement or non-nutritive sucking in newborns. Non-nutritive sucking is used to evaluate neurological function after birth, but there are no fetal indicators. The purpose of this study was to clarify the changes in the RMM clusters in fetuses at 24-39 weeks of gestation, and to investigate the relationship with the non-eye movement (NEM) period, which corresponds to the quiet sleep period after birth. Subjects included 83 normal single pregnancy cases. Fetal RMMs and eye movement (EM) were observed for 60 minutes using ultrasonic tomography and recorded as moving image files. We created time series data of eye movements and mouth movements from video recordings, and calculated RMM clusters per minute within effective observation time, RMM clusters per minute in EM period, RMM clusters per minute in NEM period, mouthing movements per cluster and ratio of number of RMM clusters per minute between NEM and EM periods and analyzed using linear regression analysis. As a result, critical points were detected in at two time points, at 32-33 weeks and 36-37 weeks of gestation, in RMM clusters per minute within the effective observation time and RMM clusters per minute in NEM period, respectively. RMM clusters in human fetuses increased from 32-33 to 36-37 weeks. This change is thought to represent fetal sleep development and central nervous system development..
37. Sonoda K¹ ², Nogami M⁴, Kodama K², Oda Y³, Kato K², Re-evaluation of preoperative endometrial smears for the cytodiagnosis of uterine leiomyosarcoma, Eur J Gynaecol Oncol, 10.31083/j.ejgo.2020.02.4950 Original Research, 41, 2, 167-170, 2020.04, Introduction: Uterine leiomyosarcoma (ULS) is a highly aggressive and lethal tumor. The absence of specific symptoms and diagnostic imaging findings makes the diagnosis of ULS challenging. Endometrial sampling reportedly has a significantly lower predictive value in diagnosing ULS compared with epithelial uterine malignancies. Materials and Methods: The authors retrospectively reviewed the preoperative endometrial cytology findings in seven of the 12 patients with ULS who were treated in this institution between 2008 and 2017. The other five patients did not have preoperative samples obtained for cytology. Results: Only one of the seven patients was originally diagnosed with a malignant tumor, with preoperative cytology showing rounded tumor cells with enlarged, irregular, hyperchromatic, and sometimes multiple nuclei, with conspicuous nucleoli in a necrotic background. After re-evaluation of the preoperative specimens, atypical cells were detected in four of the six patients who were initially deemed to have negative findings. A monomorphic population of spindle cells with slightly hyperchromatic, elongated nuclei was detected in two patients. Another two patients had rounded tumor cells with a moderate amount of basophilic cytoplasm and round or oval nuclei with minimal atypia. Conclusion: Novel diagnostic techniques are needed to accurately identify ULS in the preoperative period. Careful microscopic observation of the entire cytological specimen, together with detailed patient medical information, are essential to making a correct preoperative diagnosis of ULS..
38. Kodama K¹, Sonoda K¹, Kijima M¹, Yamaguchi S¹, Yagi H¹, Yasunaga M¹, Ohgami T¹, Onoyama I¹, Kaneki E¹, Okugawa K¹, Yahata H¹, Ohishi Y¹, Oda Y¹, Kato K¹, Retrospective Analysis of Treatment and Prognosis for Uterine Leiomyosarcoma: 10-year Experience of a Single Institute, Asia Pac J Clin Oncol, 10.1111/ajco.13286, 16, 2, e63-e67, 2020.04, AIM: Uterine leiomyosarcoma (ULMS) is a highly aggressive and lethal disease. This malignancy remains the most common type of uterine sarcoma, affecting approximately 0.4/100 000 women each year. Our aim was to assess the treatment and prognosis of ULMS patients.

METHODS: A total of 14 patients were treated at our institution between January 2008 and July 2017. We retrospectively analyzed their clinicopathological variables, treatment and prognosis.

RESULTS: The median patient age was 63 years (range, 35-83 years). The largest group of patients had stage IB disease (stage IB, n = 8; IIB, n = 2; IIIB, n = 1; IVB, n = 3) and the largest group by histological subtype was ordinary (ordinary, n = 11; myxoid, n = 2; epithelioid, n = 1). Total abdominal hysterectomy and bilateral salpingo-oophorectomy were performed for all patients, with additional surgical procedures (e.g., tumor resection, lymphadenectomy) performed if necessary. Twelve patients received adjuvant chemotherapy (ACT) consisting of gemcitabine and docetaxel. Ten patients experienced recurrence and received multidisciplinary therapies, including tumor resection, chemotherapy, radiation and targeted therapies. The median observation period was 17 months (range, 5-75 months), and 11 patients were alive (without disease, n = 5; with disease, n = 6). Intriguingly, five of eight stage IB patients who received postoperative ACT were alive without disease.

CONCLUSION: ULMS is rare but is associated with a poor prognosis, even if multidisciplinary therapies are administered. However, ACT appears to be effective in improving the prognosis of patients with stage IB disease..
39. Hamada N, Hamazaki N, Shimamoto S, Hikabe O, Nagamatsu G, Takada Y, Kato K, Hayashi K, Germ cell-intrinsic effects of sex chromosomes on early oocyte differentiation in mice, PLoS Genet, 10.1371/journal.pgen, 16, 3, e1008676-e1008676, 2020.03, A set of sex chromosomes is required for gametogenesis in both males and females, as represented by sex chromosome disorders causing agametic phenotypes. Although studies using model animals have investigated the functional requirement of sex chromosomes, involvement of these chromosomes in gametogenesis remains elusive. Here, we elicit a germ cell-intrinsic effect of sex chromosomes on oogenesis, using a novel culture system in which oocytes were induced from embryonic stem cells (ESCs) harboring XX, XO or XY. In the culture system, oogenesis using XO and XY ESCs was severely disturbed, with XY ESCs being more strongly affected. The culture system revealed multiple defects in the oogenesis of XO and XY ESCs, such as delayed meiotic entry and progression, and mispairing of the homologous chromosomes. Interestingly, Eif2s3y, a Y-linked gene that promotes proliferation of spermatogonia, had an inhibitory effect on oogenesis. This led us to the concept that male and female gametogenesis appear to be in mutual conflict at an early stage. This study provides a deeper understanding of oogenesis under a sex-reversal condition..
40. Yagi H¹, Onoyama I¹, Asanoma K¹, Hori E¹, Yasunaga M¹, Kodama K¹, Kijima M¹, Ohgami T¹, Kaneki E¹, Okugawa K¹, Yahata H¹, Kato K¹, Gα13-mediated LATS1 down-regulation contributes to epithelial-mesenchymal transition in ovarian cancer., FASEB J, doi: 10.1096/fj.201901278R, 33, 12, 13683-13694, 2019.12, Gα13, a heterotrimeric G-protein of the Gα12/13 subfamily, is associated with aggressive phenotypes in various human cancers. However, the mechanisms by which Gα13 promotes cancer progression have not been fully elucidated. Here, we demonstrate that the activation of Gα13 induces epithelial-mesenchymal transition in ovarian cancer (OvCa) cells through down-regulation of large tumor suppressor kinase (LATS) 1, a critical component of the Hippo signaling pathway. A synthetic biology approach using a mutant GPCR and chimeric G-protein revealed that Gα13-regulated phosphorylation of LATS1 at serine 909 within its activation loop induced recruitment of the itchy E3 ubiquitin protein ligase to trigger LATS1 degradation. Our findings uncover novel mechanisms through which Gα13 activation induces dysregulation of the Hippo signaling pathway, which leads to aggressive cancer phenotypes, and thereby identify a potential target for preventing the metastatic spread of OvCa.-Yagi, H., Onoyama, I., Asanoma, K., Hori, E., Yasunaga, M., Kodama, K., Kijima, M., Ohgami, T., Kaneki, E., Okugawa, K., Yahata, H., Kato, K. Gα13-mediated LATS1 down-regulation contributes to epithelial-mesenchymal transition in ovarian cancer..
41. Yahata H¹, Sonoda K¹, Okugawa K¹, Yagi H¹, Ohgami T¹, Yasunaga M¹, Onoyama I¹, Kaneki E¹, Asanoma K¹, Kato K¹, Survey of the desire to have children and engage in sexual activity after trachelectomy among young Japanese women with early-stage cervical cancer, J Obstet Gynaecol Res, 10.1111/jog.14099, 2019.08, AIM: To evaluate how the desire to have children and engage in sexual activity change after trachelectomy in Japanese women with early-stage cervical cancer who strongly desired to have children before surgery.
METHODS: Desire to have children, coital pain, fear of sexual intercourse, sexual activity frequency and libido were assessed in cervical cancer patients who received follow-up after trachelectomy. An anonymous questionnaire survey was conducted via informed consent.
RESULTS: Of the 151 patients who underwent trachelectomy at Kyushu University Hospital between 2005 and 2015, 46 patients were evaluated; the response rate was 30%. The desire to have children disappeared in 13 of 46 (28%) patients, and 14 (30%) patients experienced increased coital pain. Moreover, 19 (41%) patients experienced fear of sexual intercourse, and sexual frequency decreased in 24 (52%) patients.
CONCLUSION: Trachelectomy is an important fertility-sparing surgical method; however, this study revealed loss of the desire to have children and/or to engage in sexual activity in some patients after surgery. Counseling about these issues is important and should be addressed..
42. Yamaguchi Y¹’², Tayama C¹, Tomikawa J¹, Akaishi R³, Kamura H¹, Matsuoka K⁴’⁵, Wake N², Minakami H⁶, Kato K², Yamada T⁷, Nakabayashi K⁸, Hata K⁹, Placenta-specific epimutation at H19-DMR among common pregnancy complications: its frequency and effect on the expression patterns of H19 and IGF2, Clin Epigenetics, 10.1186/s13148-019-0712-3, 11, 1, 113-113, 2019.08, BACKGROUND: H19 and IGF2 genes are imprinted and involved in regulating fetal and placental growth. The H19 differentially methylated region (DMR) is paternally methylated and maternally unmethylated and regulates the imprinted expression of H19 and IGF2. Epimutation at the H19-DMR in humans results in congenital growth disorders, Beckwith-Wiedemann and Silver-Russell syndromes, when erroneously its maternal allele becomes methylated and its paternal allele becomes unmethylated, respectively. Although H19 and IGF2 have been assessed for their involvement in pregnancy complications including fetal growth restriction (FGR) and pregnancy-induced hypertension (PIH)/hypertensive disorder of pregnancy (HDP) intensively in the last decade, it is still not established whether epimutation at the H19-DMR in the placenta results in pathogenic conditions in pregnancy. We aimed to assess the frequency of H19-DMR epimutation and its effects on the allelic expression patterns of H19 and IGF2 genes among normal and abnormal pregnancy cases.
RESULTS: We enrolled two independently collected sets of placenta samples from normal pregnancies as controls and common pregnancy complications, FGR and PIH (HDP). The first set consisted of 39 controls and 140 FGR and/or PIH cases, and the second set consisted of 29 controls and 62 cases. For these samples, we initially screened for DNA methylation changes at H19-DMR and IGF2-DMRs by combined bisulfite restriction analysis, and further analyzed cases with methylation changes for their allelic methylation and expression patterns. We identified one case each of FGR and PIH showing hypomethylation of H19-DMR and IGF2-DMRs only in the placenta, but not in cord blood, from the first case/control set. For the PIH case, we were able to determine the allelic expression pattern of H19 to be biallelically expressed and the H19/IGF2 expression ratio to be highly elevated compared to controls. We also identified a PIH case with hypomethylation at H19-DMR and IGF2-DMRs in the placenta from the second case/control set.
CONCLUSIONS: Placental epimutation at H19-DMR was observed among common pregnancy complication cases at the frequency of 1.5% (3 out of 202 cases examined), but not in 68 normal pregnancy cases examined. Alteration of H19/IGF2 expression patterns due to hypomethylation of H19-DMR may have been involved in the pathogenesis of pregnancy complications in these cases..
43. Ikeda Y¹, Uemura Y², Asai-Sato M¹, Nakao T¹, Nakajima T¹, Iwata T³, Akiyama A⁴, Satoh T⁴, Yahata H⁵, Kato K⁵, Maeda D⁶, Aoki D³, Kawana K¹, Safety and efficacy of mucosal immunotherapy using human papilliomavirus (HPV) type 16 E7-expressing Lactobacillus-based vaccine for the treatment of high-grade squamous intraepithelial lesion (HSIL): the study protocol of a ran-domized placebo-controlled clinical trial (MILACLE study), Japanese Journal of Clinical Oncology, 10.1093/jico/hyz095, 2019,1-4, 2019.07, We developed an HPV16 E7-expressing Lactobacillus-based therapeutic vaccine, IGMKK16E7, to elicit mucosal E7-specific TH1 cellular immune responses. This study aims to examine the safety and clinical efficacy of IGMKK16E7 on HPV16-positive high-grade squamous intraepithelial lesion (HSIL). This is a multicenter, placebo-controlled, double-blind randomized phase I/II trial to test the safety and efficacy of IGMKK16E7 against HPV16-positive HSIL. The groups will include placebo, low-dose (0.5 g/day), middle-dose (1 g/day), and high-dose (1.5 g/day) IGMKK16E7. The target sample size will be 41 patients per group, and our data on our former agent, GLBL101c, were used to calculate sample size for 70% power and an α level = 0.05. The primary endpoint is IGMKK16E7 safety and pathological regression at week 16, and the secondary endpoints are cytological regression and HPV16 E7 immunological response. This study protocol has been approved by the Japanese Pharmaceuticals and Medical Devices Agency. Patient enrollment will begin in May 2019..
44. Asanoma K¹, Hori E¹, Yoshida S¹, Yagi H¹, Onoyama I¹, Kodama K¹, Yasunaga M¹, Ohgami T¹, Kaneki E¹, Okugawa K¹, Yahata H¹, Kato K¹, Mutual suppression between BHLHE40/BHLHE41 and the MIR301B-MIR130B cluster is involved in epithelial-to-mesenchymal transition of endometrial cancer cells, Oncotarget, 10.18632/oncotarget.27061, 45, 10, 4640-4654, 2019.07, BHLHE40 and BHLHE41 (BHLHE40/41) are basic helix-loop-helix type transcription factors involved in multiple cell activities including epithelial-to-mesenchymal transition (EMT). However, the expression mechanism of BHLHE40/41 in EMT remains unclear. In the present study, we showed that the expression levels of BHLHE40/41 were negatively correlated with those of the microRNA (MIR) 130 family in endometrial cancer (EC) specimens. Our in vitro assays indicated that the expression of BHLHE40/41 was suppressed directly by the MIR130 family in a 3'-untranslated region-mediated manner. In EC cells, the MIR130 family promoted EMT and tumor cell invasion by suppressing the expression of BHLHE40/41. We identified the critical promoter region of the MIR301B-MIR130B cluster for its basal transcription by the transcription factor, SP1. We also found that BHLHE40/41 suppressed the expression of MIR301B and MIR130B, and we identified a binding site in the promoter region for BHLHE40/41. This study is the first to report that BHLHE40/41 and the MIR301B-MIR130B cluster suppressed each other to regulate EMT and invasion of EC cells. We propose that BHLHE40/41 and the MIR130 family are excellent markers to predict the progression of EC cases, and that molecular therapy targeting the MIR130 family-BHLHE40/41 axis may effectively control EC extension..
45. Inoue S¹,², Ito H¹, Hosono S¹, Hori M¹, Matsuda T¹, Mizuno M¹, Kato K², Matsuo K¹, Net Survival of Elderly Patients with Gynecological Cancer Aged Over 75 Years in 2006-2008, Asian Pacific journal of cancer prevention, 10.31557/APJCP.2019.20.2.437, 20, 2, 437-442, 2019.02, BACKGROUND: The number of elderly patients with gynecological cancer in Japan is increasing in line with the aging of society. However, little has been reported on the survival of elderly patients aged 75 or older with gynecological cancer in Japan.
METHODS: To clarify survival in women aged 75 years or older with gynecological cancer, we analyzed data of 4,089 gynecological cancer cases (cervical cancer, 1,309 cases; endometrial cancer, 1,319 cases; and ovarian cancer, 1,461 cases) in patients aged 75 or older from 21 population-based cancer registries in Japan, diagnosed in 2006-2008. We calculated the net survival (NS) of younger (75-79 years old), older (80-84 years old) and the oldest age group (85-99 years old). We also calculated NS stratified by extent of disease and histological type.
RESULTS:Five-year NS of cervical cancer patients was 54.5% in the younger age group, 40.8% in the older age group and 28.2% in the oldest age group. Five-year NS of endometrial cancer patients was 64.5%, 51.6% and 39.0% in the younger, older and oldest age groups, respectively. Five-year NS of ovarian cancer was 34.7%, 18.8% and 8.3%, respectively.
CONCLUSION:We estimated NS in elderly patients aged 75 years or older with gynecological cancers in Japan using data from population-based cancer registries..
46. Fujita Y¹, Nakanishi TO², Sugitani M², Kato K², Placental Elasticity as a New Non-invasive Predictive Marker of Pre-eclampsia., Ultrasound in Med. & Biol, 10.1016/j.ultrasmedbio.2018.09.007 , 45, 1, 93-97, 2019.01, Point shear wave elastography is an ultrasonography technique used to evaluate tissue elasticity. We examined whether placental elasticity is useful for predicting the onset of pre-eclampsia. Two hundred twenty-one participants were divided into two groups: one group at low risk (n = 185) and the other at high risk (n = 36) for pre-eclampsia. The two groups were compared with respect to shear wave velocity (SWV) of the placenta. Use of SWV as a predictor of pre-eclampsia was also investigated by creating a receiver operating characteristic (ROC) curve. The ROC curve was used to set a cutoff SWV value for predicting pre-eclampsia. The SWV of the high-risk group was significantly higher than that of the low-risk group (p < 0.001). Thirteen participants developed pre-eclampsia after SWV measurements, and the SWVs of these participants were significantly higher than those of participants in who pre-eclampsia did not develop. The cutoff value and area under the ROC curve were 1.188 m/s and 0.9118, respectively. Placental elasticity was significantly increased even before the onset of pre-eclampsia onset and, thus, may be a parameter used to predict the onset of pre-eclampsia..
47. Morisaki N¹, Nagata C², Yasuo S³, Morokuma S⁴, Kato K⁵, Sanefuji M⁴, Shibata E⁶, Tsuji M⁷, Senju A⁶, Kawamoto T⁷, Ohga S⁸, Kusuhara K⁹, Japan Environment and Children’s Study Group, Optimal protein intake during pregnancy for reducing the risk of fetal growth restriction: the Japan Environment and Children's Study, Br J Nutr, 10.1017/S000711451800291X, 120, 12, 1432-1440, 2018.12, Clinical trials show that protein supplement increases infant size in malnourished populations; however, epidemiological studies in high-income countries have reported mixed results. Although these findings suggest a non-linear relationship between maternal macronutrient intake and fetal growth, this relationship has not been closely examined. We assessed the association between maternal protein intake and fetal growth among 91 637 Japanese women with singletons in a nation-wide cohort study using validated FFQ. The respondents answered the FFQ twice, once during early pregnancy (FFQ1; 16·3 (sd 6·0) weeks), and second during mid-pregnancy (FFQ2, 28·1 (sd 4·1) weeks). Daily energy intake and percentage energy from protein, fats and carbohydrates were 7477 (sd 2577) kJ and 13·5 (sd 2·0), 29·5 (sd 6·5) and 55·3 (sd 7·8) %, respectively, for FFQ1, and 7184 (sd 2506) kJ and 13·6 (sd 2·1), 29·8 (sd 6·6) and 55·3 (sd 7·9) %, respectively, for FFQ2. The average birth weight was 3028 (sd 406) g, and 6350 infants (6·9 %) were small for gestational age (SGA). In both phases of the survey, birth weight was highest and the risk of SGA was lowest when the percentage energy from protein was 12 %, regardless of whether isoenergetic replacement was with fat or carbohydrates. Furthermore, when protein density in the maternal diet was held constant, birth weight was highest when 25 % of energy intake came from fat and 61 % came from carbohydrates during early pregnancy. We found maternal protein intake to have an inverse U-curve relationship with fetal growth. Our results strongly suggest that the effect of protein on birth weight is non-linear, and that a balanced diet fulfilling the minimum requirement for all macronutrients was ideal for avoiding fetal growth restriction..
48. Egashira K¹, Hiasa K¹, Yokota N¹, Kawamura T¹, Matsushita T¹, Okugawa K¹, Yahata H¹, Sonoda K¹, Kato K¹, Infertility after abdominal trachelectomy, Acta Obstet Gynecol Scand, doi: 10.1111/aogs.13429, 97, 11, 1358-1364, 2018.11, INTRODUCTION: Despite numerous reports on pregnancy outcomes after trachelectomy, there are few descriptions of fertility treatment after trachelectomy. Moreover, little is known about the differences in fertility outcomes between various radical trachelectomy procedures. The purpose of this report was to clarify the infertility problems that occur in patients who have previously undergone an abdominal trachelectomy.
MATERIAL AND METHODS: We retrospectively investigated the medical records of 37 patients who received fertility treatments or were evaluated for menstrual disorders after trachelectomies in our institution between 2012 and 2016.
RESULTS: Twenty-two of 37 patients had complications which affected fecundity. Six patients had cervical stenosis requiring surgical dilation, 4 had ovarian insufficiency, and 14 had Asherman's syndrome.
CONCLUSIONS: In spite of efforts to preserve fertility, some patients have severe complications after trachelectomy, such as Asherman's syndrome, resulting in infertility. Clinicians should pay careful attention to the status of the endometrial cavity after trachelectomy..
49. Sato Y¹, Hidaka N¹, Nakano T¹, Kido S¹, Hachisuga M¹, Fujita Y¹, Kato K¹, Efficacy of an Emergency Cervical Cerclage Using Absorbable Monofilament Su-tures, Journal of Pregnancy 2018:4049792, 10.1155/2018/4049792, 2018.11, Introduction: Although nonabsorbable woven tape has been widely used for cervical cerclage, technical difficulties that can occur with an effaced cervix because of the thickness of the tape, and the risks of local infection are two major concerns. This study investigated perinatal outcomes of pregnancies involving an emergency cervical cerclage using absorbable monofilament polydioxanone sutures, which is a narrow thread and protects against bacterial infection.
Materials and Methods: We performed a chart review of patients who underwent emergency McDonald cerclage with polydioxanone sutures at our institution between 2007 and 2015. Gestational age at delivery, duration between cerclage and delivery, and neonatal prognosis were evaluated as primary outcomes.
Results:Among the 23 patients (18 singleton and five twin pregnancies) evaluated, ultrasound-indicated (progressive cervical length shortening) were eight (35%) and physical examination-indicated (fetal membranes that prolapsed into the vagina or dilated cervix) were 15 patients (65%). The median gestational age at cerclage was 22+3 weeks (range, 17+5 to 25+3 weeks). Postoperative spontaneous abortion occurred in only one patient. The median gestational age at delivery was 32+5 weeks (range, 20+5 to 40+6 weeks). Extremely preterm delivery before 28 weeks of gestation occurred in four (17%) cases. Full-term delivery was achieved in 10 (42%) cases. The duration between cerclage and delivery ranged from 5 to 136 days (median, 77 days). Except for one case of spontaneous abortion, all newborns survived till hospital discharge.
Conclusions:Although our series included some patients at high risk for spontaneous abortion and preterm delivery, satisfactory prolongation and favorable neonatal outcomes were achieved for most patients by using absorbable monofilament sutures, thus suggesting the efficacy of this type of suture for emergency cervical cerclage..
50. Morokuma S¹’², Michikawa T³, Kato K⁴’⁵, Sanefuji M⁴’⁶, Shibata E⁷’⁸, Tsuji M⁹, Senju A⁷'¹⁰, Kawamoto T⁷’⁹, Ohga S⁴’⁶, Kusuhara K⁷’¹⁰, Non-reassuring foetal status and neonatal irritability in the Japan Environment and Children's Study: A cohort study., Sci Rep, 10.1038/s41598-018-34231-y, 8, 1, 15853-15853, 2018.10, The aim of this study was to investigate whether non-reassuring foetal status (NRFS) affected an infant's temperament, or if the temperament formed prenatally resulted in an excessive heart rate reaction that was diagnosed as NRFS. We examined the correlation between NRFS and difficulty in holding a baby, and the amount of crying in the one month after birth, which was considered an indicator of the newborn's temperament. We divided the cases with NRFS into positive NRFS and false positive NRFS. NRFS was associated with bad mood, frequent crying for a long duration, and intense crying. After adjustment for other covariates, NRFS was associated with bad mood (odds ratio, OR = 1.15, 95% confidence interval, CI = 1.00-1.33), and intense crying (1.12, 1.02-1.24). In the multi-variable model, positive and false positive NRFS were not clearly associated with neonatal irritability. When stratified by parity, NRFS and false positive NRFS were likely to be positively associated with neonatal irritability in parous women. The clear association between NRFS and intense crying was observed in parous women (multi-variable adjusted OR = 1.46, 95% CI = 1.16-1.83), but not in nulliparae (1.01, 0.91-1.12) (p for effect modification <0.01). Similarly, increased odds of intense crying associated with false positive NRFS were only found in parous women (multi-variable adjusted OR = 1.40, 95% CI = 1.09-1.81) (p for effect modification = 0.03). There was no association observed between positive NRFS and irritability; therefore, NRFS has no effect on an infant's temperament..
51. Okugawa K¹, Sonoda K¹, Ohgami T¹, Yasunaga M¹, Kaneki E¹, Yahata H¹, Kato K¹, Pelvic abscess:A late complication of abdominal trachelectomy for cervical cancer, J Obstet Gynaecol, 10.1111/jog.13814, 2018.09, AIM:Only a few reports of pelvic abscess as a late complication of trachelectomy have been published to date. To evaluate the cases of pelvic abscess as a late complication of abdominal trachelectomy for cervical cancer.
METHODS: In June 2005, we began a clinical trial of abdominal trachelectomy at our institution. Written informed consent was obtained from all patients. We retrospectively reviewed the medical records of patients who underwent trachelectomy and extracted the data of patients who experienced pelvic abscess as a late complication.
RESULTS: From June 2005 to September 2017, we performed 181 trachelectomies at our institution. In total, 15 pelvic abscesses occurred in 12 of these patients more than 1 month after trachelectomy. The median postoperative period before the onset of pelvic abscess was 51 months (range, 1-104 months). Among the 15 cases, abscess formed in the uterine adnexa in 12, in a pelvic lymphocyst in two, and in the uterus in one. Abscess drainage was performed in six cases. Three patients underwent laparotomy with salpingo-oophorectomy.
CONCLUSION: It is possible that not only surgical removal of the uterine cervix but also the use of nonabsorbable suture in cervical cerclage and placement of an intrauterine device triggered post-trachelectomy infection. Pelvic abscess can occur as a late complication of abdominal trachelectomy..
52. Kitawaki J¹, Kato K², Annual report of the Women's Health Care Committee, Japan Society of Obstetrics and Gynecology, 2018., J Obstet Gynaecol Res, 10.1111/jog.13803, 2018.09, Since 2010, the Women's Health Care Committee has addressed important issues linked to women's quality of life. The five agendas this committee has engaged this academic year are: (i) clinical study of urinary complications in perinatal and postpartum women's health care; (ii) education on infectious diseases prophylaxis in obstetrics and gynecology in Japan; (iii) educational activities regarding management guidelines for a female athlete's health care; (iv) training program for women's healthcare advisers; and (v) pregnancy- and lactation-associated osteoporosis (PLOP). The detailed activities of the five subcommittees are described herein. This report is based on the Japanese version of our annual report (Acta Obst Gynaec Jpn 2018; 70: 1538-1546), which publicizes the activities of our committee..
53. Yahata H, Kobayashi H, Sonoda K, Kodama K, Yagi H, Yasunaga M, Ohgami T, Onoyama I, Kaneki E, Okugawa K, Baba S, Isoda T, Ohishi Y, Oda Y, Kato K, Prognostic outcome and complications of sentinel lymph node navigation surgery for early-stage cervical cancer., Int J Clin Oncol, 10.1007/s10147-018-1327-y, 2018.08, BACKGROUND: To evaluate the prognostic outcome and surgical complications in patients with early-stage cervical cancer who underwent sentinel node navigation surgery (SNNS) for hysterectomy or trachelectomy.
METHODS: A total of 139 patients who underwent SNNS using 99mTc phytate between 2009 and 2015 were evaluated. No further lymph node dissection was performed when intraoperative analysis of the sentinel lymph nodes (SLNs) was negative for metastasis. We compared the surgical complications between the SNNS group and 67 matched patients who underwent pelvic lymph node dissection (PLND) after SLN mapping between 2003 and 2008. We also examined the clinical outcomes in the SNNS group.
RESULTS: The mean number of detected SLNs was 2.5 per patient. Fourteen of the 139 patients in the SNNS group underwent PLND based on the intraoperative SLN results. The amount of blood loss, the operative time, and the number of perioperative complications were significantly less in the SNNS group than in the matched PLND group. There was no recurrence during a follow-up period ranging from 2 to 88 months (median 40 months) in the SNNS group.
CONCLUSIONS: Using SNNS for early-stage cervical cancer is safe and effective and does not increase the recurrence rate. A future multicenter trial is warranted..
54. Fujita Y, Kiyokoba R, Yumoto Y, Kato K, Max dD/Dt: A Novel Parameter to Assess Fetal Cardiac Contractility and a Sub-stitute for Max dP/Dt., Ultrasound Med Biol, 10.1016/j.ultrasmedbio.2018.02.001, 44, 7, 1433-1438, 2018.07, Aortic pulse waveforms are composed of a forward wave from the heart and a reflection wave from the periphery. We focused on this forward wave and suggested a new parameter, the maximum slope of aortic pulse waveforms (max dD/dt), for fetal cardiac contractility. Max dD/dt was calculated from fetal aortic pulse waveforms recorded with an echo-tracking system. A normal range of max dD/dt was constructed in 105 healthy fetuses using linear regression analysis. Twenty-two fetuses with suspected fetal cardiac dysfunction were divided into normal and decreased max dD/dt groups, and their clinical parameters were compared. Max dD/dt of aortic pulse waveforms increased linearly with advancing gestational age (r = 0.93). The decreased max dD/dt was associated with abnormal cardiotocography findings and short- and long-term prognosis. In conclusion, max dD/dt calculated from the aortic pulse waveforms in fetuses can substitute for max dP/dt, an index of cardiac contractility in adults..
55. Morokuma S¹, Maehara K², Okawa H², Kato K², Mine Y³, Nakauchi S³, Simplified Wide-Range Ultrasonic Measurements Using the Sensor Three-Dimensional System, J Med Ultrasound, 10.4103/JMU.JMU_17_18, 26, 2, 100-102, 2018.06, We developed a simplified three-dimensional ultrasonic device that can scan a wide area and performed measurements in the scanned area. The system is more compact than magnetic resonance imaging (MRI) systems and can measure random cross sections by acquiring volume data over a wide range through freehand scanning with a magnetic sensor unit that detects the transducer position. The system was applied successfully to a case with a huge myoma. Our system, in ways similar to computed tomography or MRI systems, can support both the objective understanding of the pathology of huge tumors and follow-up determinations of tumor diameters at arbitrary cross sections in the volume data..
56. Yasutake N¹, Ohishi Y¹, Taguchi K², Hiraki Y³, Oya M³, Oshiro Y⁴, Mine M⁵, Iwasaki T¹, Yamamoto H¹, Kohashi K¹, Sonoda K⁶, Kato K⁶, Oda Y¹, Insulin-like growth factor II messenger RNA-binding protein-3 is an independ-ent prognostic factor in uterine leiomyosarcoma, Histopathology, 10.1111/his.13422, 72, 5, 739-748, 2018.04, AIMS: The aim of this study was to identify the prognostic factors of uterine leiomyosarcoma (ULMS).
METHODS AND RESULTS: We reviewed 60 cases of surgically resected ULMSs and investigated conventional clinicopathological factors, together with the expression of insulin-like growth factor II messenger RNA-binding protein-3 (IMP3), hormone receptors and cell cycle regulatory markers by immunohistochemistry. Mediator complex subunit 12 (MED12) mutation analysis was also performed. Univariate analyses revealed that advanced stage (P < 0.0001), older age (P = 0.0244) and IMP3 expression (P = 0.0011) were significant predictors of a poor outcome. Multivariate analysis revealed advanced stage (P < 0.0001) and IMP3 (P = 0.0373) as independent predictors of a poor prognosis. Expressions of cell cycle markers and hormone receptors, and MED12 mutations (12% in ULMSs) were not identified as prognostic markers in this study.
CONCLUSIONS: IMP3 expression in ULMS could be a marker of a poor prognosis..
57. Sakai A¹'², Matsuda T¹, Doi H¹, Nagaishi Y¹, Kato K², Nakashima K³, Ectopic neurogenesis induced by prenatal antiepileptic drug exposure augments seizure susceptibility in adult mice, Proc Natl Acad Sci U S A, 10.1073/pnas.1716479115, 115, 16, 4270-4275, 2018.04, Epilepsy is a neurological disorder often associated with seizure that affects ∼0.7% of pregnant women. During pregnancy, most epileptic patients are prescribed antiepileptic drugs (AEDs) such as valproic acid (VPA) to control seizure activity. Here, we show that prenatal exposure to VPA in mice increases seizure susceptibility in adult offspring through mislocalization of newborn neurons in the hippocampus. We confirmed that neurons newly generated from neural stem/progenitor cells (NS/PCs) are integrated into the granular cell layer in the adult hippocampus; however, prenatal VPA treatment altered the expression in NS/PCs of genes associated with cell migration, including CXC motif chemokine receptor 4 (Cxcr4), consequently increasing the ectopic localization of newborn neurons in the hilus. We also found that voluntary exercise in a running wheel suppressed this ectopic neurogenesis and countered the enhanced seizure susceptibility caused by prenatal VPA exposure, probably by normalizing the VPA-disrupted expression of multiple genes including Cxcr4 in adult NS/PCs. Replenishing Cxcr4 expression alone in NS/PCs was sufficient to overcome the aberrant migration of newborn neurons and increased seizure susceptibility in VPA-exposed mice. Thus, prenatal exposure to an AED, VPA, has a long-term effect on the behavior of NS/PCs in offspring, but this effect can be counteracted by a simple physical activity. Our findings offer a step to developing strategies for managing detrimental effects in offspring exposed to VPA in utero..
58. Yahata H, Sonoda K, Yasunaga M, Ogami T, Kawano Y, Kaneki E, Okugawa K, Tsunehisa Kaku, Kato K, Surgical treatment and outcome of early invasive adenocarcinoma of the uterine cervix (FIGO stage IA1)., Asia Pac J Clin Oncol 2017, 10.1111/ajco.12691, 14, 2, e50-e53, 2018.04, AIM: To investigate the surgical outcome of FIGO stage IA1 cervical adenocarcinoma.
METHODS: Between 2005 and 2011, 12 patients from Kyushu University Hospital had cervical adenocarcinoma, with a tumor depth of less than 3 mm and a horizontal width of less than 7 mm (FIGO stage IA1), diagnosed by cervical conization. All patients underwent simple hysterectomy or simple trachelectomy with pelvic lymphadenectomy.
RESULTS: The mean patient age was 34 years (range, 26-70 years). The median follow-up period was 70.5 months (range, 26-99 months). No pelvic lymph-node metastasis was seen, and no patient experienced disease recurrence.
CONCLUSION:Early invasive cervical adenocarcinoma with a depth of invasion of 3 mm or less and a horizontal spread of 7 mm or less has little potential for nodal metastasis or recurrence. Therefore, simple hysterectomy or trachelectomy, without lymphadenectomy, might be an alternative treatment option for stage IA1 cervical adenocarcinoma..
59. Ohmaru-Nakanishi T¹, Asanoma K², Fujikawa M¹, Fujita Y¹, Yagi H¹, Onoyama I¹, Hidaka N¹, Sonoda K¹, Kato K¹, Fibrosis in Preeclamptic Placentas Is Associated with Stromal Fibroblasts Acti-vated by the Transforming Growth Factor Beta 1 (TGFB1) Signaling Pathway., Am J Pathol, 10.1016/j.ajpath.2017.11.008, 188, 3, 683-695, 2018.03, Although fibrosis is one of the most prominent pathologic features of preeclamptic (PE) placentas, its mechanism remains largely unknown. Consistent with previous reports, we observed overexpression of collagen; actin, α2, smooth muscle, aorta; connective tissue growth factor; and fibronectin in PE placentas compared with control ones. To investigate the mechanism of fibrosis in PE placentas, placental fibroblasts were isolated from PE placentas or normal pregnancies at delivery. The expression of fibrosis-related factors in fibroblasts was evaluated by real-time RT-PCR, Western blotting, enzyme-linked immunosorbent assay, and gene microarrays. An in vitro collagen gel contraction assay was also performed. Fibroblasts isolated from PE placentas showed higher expression levels of fibrosis-related factors compared with those from control ones. Global gene expression profiling of PE fibroblasts was contrasted with that of control ones and indicated an intimate association with transforming growth factor-β1 (TGF-β1) signaling. Furthermore, the PE fibroblasts expressed abundant phosphorylated SMAD family member 2 and showed higher expression levels of target genes of TGF-β1 signaling compared with the control ones. The PE fibroblasts also had a greater ability to contract compared with the control ones. Contractility also depended on TGF-β1 signaling. Our results suggest that TGF-β1 signaling is activated in the fibroblasts in PE placentas and that these active fibroblasts contribute to fibrosis..
60. Muraoka M¹, Nagata H², Hirata Y¹, Uike K¹, Terashi E¹, Morihana E¹, Ochiai M¹, Fujita Y³,Kato K³, Yamamura K¹, Ohga S¹, High incidence of progressive stenosis in aberrant left subclavian artery with right aortic arch., Heart Vessels, 10.1007/s00380-017-1056-6, 33, 3, 309-315, 2018.03, Right aortic arch with aberrant left subclavian artery (RAA/aLSCA) is a rare aortic arch anomaly. The clinical association of aLSCA stenosis with RAA/aLSCA has not yet been fully elucidated. The aim of this study was to investigate the diagnosis, incidence, management and outcome of aLSCA stenosis in infants with prenatally diagnosed RAA/aLSCA. Ten fetuses who were diagnosed as having RAA/aLSCA in Kyushu University Hospital between January 2011 and December 2014 were enrolled. The maternal and child medical records were reviewed to investigate sex, gestational age at the fetal diagnosis, gestational age and body weight at birth, the findings of computed tomography (CT), Doppler ultrasonography of the vertebral artery and angiography, and the complications and outcomes of aLSCA stenosis. In 8 of 10 patients, aLSCA stenosis was identified on the first CT examination after birth. No patients had dysphagia or respiratory distress. The stenosis spontaneously resolved in 3 patients. In 4 of the 5 remaining patients, aLSCA stenosis progressed, including one case in which complete occlusion occurred-the case was associated with retrograde flow from the left vertebral artery supplying the distal LSCA. Balloon angioplasty was successfully used to treat stenosis in two cases. The subclavian steal phenomenon and developmental problems were not observed in any patients. aLSCA stenosis was identified in 80% of patients with RAA/aLSCA after birth. The early detection and elective treatment of stenotic lesions may be required to prevent complete occlusion during the development of the cardiovascular and cerebrovascular systems..
61. Hidaka N , Kido S, Sato Y, Murata M, Fujita Y, Kato K, Thoracoamniotic shunting for fetal pleural effusion with hydropic change using a double-basket catheter: An insight into the preoperative determinants of shunt-ing efficacy., Eur J Obstet Gynecol Reprod Biol, 10.1016/j.ejogrb.2017.12.008, 221, 34-39, 2018.02, OBJECTIVES: Although the efficacy of thoracoamniotic shunting (TAS) for fetal hydrothorax is well-recognized, the coexistence of hydrops fetalis is still a clinical challenge. The preoperative determinants of shunting efficacy are not fully understood. In this study, we aimed to investigate the perinatal and postnatal outcomes of hydrops fetalis with pleural effusion treated by TAS using a double-basket catheter, and to discuss the preoperative factors predictive of patients who will benefit from TAS.

STUDY DESIGN: We conducted a retrospective study in hydropic fetuses with pleural effusion treated by TAS between 2007 and 2015. We extracted information regarding postnatal survival and pretherapeutic sonographic findings, including skin-edema thickness, pleural-effusion pocket size, and Doppler readings.

RESULTS: Twelve subjects underwent TAS at a median gestational age of 29+5 weeks (range, 25+5-33+2 weeks). Skin edema disappeared or regressed in 7. Three experienced early neonatal death and the other 9 ultimately survived after a live birth at a median gestational age of 33+4 weeks (range, 29+1-38+2 weeks). All surviving children, except for 1, had a pretherapeutic pleural-effusion pocket greater than the precordial-edema thickness. All 3 children that died had precordial-edema thickness equal to or greater than the size of the pleural-effusion pocket.

CONCLUSIONS: We achieved a high survival rate (75%) using the double-basket technique. A greater pretherapeutic width of skin edema compared with the pleural-effusion pocket is possibly suggestive of a treatment-resistant condition and subsequent poor postnatal outcome..
62. Inoue S¹’², Hosono S³, Ito H¹’⁴, Oze I¹, Nishino Y⁵, Hattori M⁶, Matsuda T⁷, Miyashiro I⁸, Nakayama T⁸, Mizuno M⁹, Matsuo K¹’⁴, Kato K², Tanaka H³’¹⁰, Ito Y⁸, J-CANSIS Research Group, Improvement in 5-Year Relative Survival in Cancer of the Corpus Uteri From 1993-2000 to 2001-2006 in Japan., J Epidemiol, 10.2188/jea.JE20170008, 28, 2, 75-80, 2018.02, BACKGROUND: Medical circumstances in Japanese patients with cancer of the corpus uteri have greatly changed since the late 1990s, including the introduction of concomitant therapy with taxane and platinum. We evaluated changes in survival rates for this cancer following these advances by analyzing data from population-based cancer registries in Japan.
METHODS:Data were available for 8562 cases of cancer of the corpus uteri from six prefectural cancer registries. We defined the two periods of 1993-2000 (1st period) and 2001-2006 (2nd period). Relative survival (RS) in each period was calculated to assess changes using an excess mortality model, with adjustment for age group (15-54, 55-69, and 70-99 years), extent of disease (localized, regional, and distant), and histological subtype.
RESULTS: Overall 5-year RS improved from 77.7% in the 1st period to 80.2% in the 2nd period, with an excess hazard ratio (EHR) of 0.785 (95% confidence interval [CI], 0.705-0.873). Five-year RS significantly improved in the group aged 55-69 years, in all groups by extent of disease, and in the endometrioid adenocarcinoma group. In particular, 5-year RS significantly improved in patients with endometrioid adenocarcinoma, from 84.5% to 89.7%, with an EHR of 0.698 (95% CI, 0.560-0.870).
CONCLUSION:Overall 5-year RS for cancer of the corpus uteri in Japan improved from the 1990s to early 2000s. These improvements might have been aided by the comprehensive medical development of management for this cancer, including the spread of concomitant therapy with taxane and platinum as a standard adjuvant chemotherapy in the early 2000s..
63. Sonoda K¹, Yahata H, Okugawa K, Kaneki E, Ohgami T, Yasunaga M, Baba S, Oda Y, Honda H, Kato K, Value of Intraoperative Cytological and Pathological Sentinel Lymph Node Di-agnosis in Fertility-Sparing Trachelectomy for Early-Stage Cervical Cancer, Oncology, 10.1159/000484049, 94, 2, 92-98, 2018.02, BACKGROUND AND OBJECTIVES: Trachelectomy, a fertility-sparing surgery for early-stage cervical cancer, can be performed only when there is no extrauterine extension present. Therefore, identifying the sentinel lymph nodes (SLNs) and using them to obtain an intraoperative pathologic diagnosis can provide information on the feasibility and safety of trachelectomy. Our aim was to assess the value of an intraoperative SLN diagnosis.
METHODS: We retrospectively analyzed the accuracy of intraoperative imprint cytology and frozen-section examination in 201 patients at our institution in whom trachelectomy was planned.
RESULTS: All patients could be evaluated for SLNs; a total of 610 SLNs were analyzed. Although the specificity of both imprint cytology and frozen-section examination was 100.0%, the sensitivity was only 58.6 and 65.5%, respectively. The diagnostic sensitivity was higher in 2-mm slices along the short axis than on bisection along the longitudinal axis. Imprint cytology correctly diagnosed 2 patients who had false-negative results on frozen section. The nature of the metastatic foci that caused an intraoperative false-negative diagnosis was either micrometastasis or isolated tumor cells.
CONCLUSIONS: The accuracy of intraoperative SLN diagnosis requires improvement, especially when small metastatic foci are present..
64. Benefits of an automated screening system for quality control of cervical cytology.
65. Ohmura Y¹, Morokuma S²’³,, Kato K³, Kuniyoshi Y⁴, Species-specific Posture of Human Foetus in Late First Trimester, Sci Rep, 10.1038/s41598-017-18384-w, 8, 1, 27-27, 2018.01, The ontogeny associated with the arm-hanging posture, which is considered ape-specific, remains unknown. To examine its ontogeny, we measured foetal movements of 62 human foetuses aged 10-20 gestation weeks using four-dimensional sonography. We observed that the first-trimester foetuses show this particular species-specific posture. After 11 weeks of gestation, all foetuses showed the arm-hanging posture, and the posture was most frequently observed at 14-16 weeks of gestation. Moreover, this posture often involved extension of both arms and both legs, indicating that it is not myogenic but neurogenic. Furthermore, early ontogeny suggests that it originates because of subcortical activity. Such posture extension bias and persistence indicates that vestibulospinal tract maturation involves the ontogeny of arm-hanging posture during 14-16 weeks of gestation..
66. Hidaka N¹, Sato Y², Kido S², Fujita Y², Kato K², Ductus venosus Doppler and the postnatal outcomes of growth restricted fetuses with absent end-diastolic blood flow in the umbilical arteries., Taiwanese journal of obstetrics & gynecology, 10.1016/j.tjog.2017.08.012, 56, 5, 642-647, 2017.10, OBJECTIVE: We aimed to evaluate the outcomes of growth-restricted fetuses with absent end-diastolic velocity in the umbilical arteries (UA-AEDV), and investigate the relationship between Doppler flow velocity waveforms in the ductus venosus (DV) and the clinical features.

MATERIALS AND METHODS: This was a retrospective study of growth-restricted fetuses diagnosed with UA-AEDV delivered at our institution between 2013 and 2015. The time from diagnosis of UA-AEDV to delivery, postnatal survival, and developmental prognoses were the primary outcomes. The time lag between the occurrence of UA-AEDV and an abnormal increase in the DV pulsatility index (DV-PI) were investigated. We also examined the correlation between the DV-PI values immediately before birth and umbilical cord arterial pH at birth.

RESULTS: The median gestational age at birth among the 18 subjects was 28+2 (24+0-34+6) weeks, and the observation period between the first detection of UA-AEDV and delivery ranged from 0 to 35 days with a median of 8 days. Among the 18 infants, 15 (83%) survived, among whom 2 were diagnosed with a developmental disability. Gestational age at delivery was significantly lower in the poor outcome group. A positive correlation (correlation coefficient, 0.68) was observed between the umbilical artery pH and the last measured DV-PI.

CONCLUSION: The time interval from initial detection of UA-AEDV to delivery is highly variable, and it is reasonable to manage these growth-restricted fetuses with UA-AEDV expectantly with careful surveillance for fetal well-being. Specifically, Doppler DV analysis is clinically valuable for their evaluation..
67. Setoguchi K, Cui L, Hachisuka N, Obchoei S, Shinkai K, Hyodo F, Kato K, Wada F, Yamamoto T, Harada-Shiba M, Obika S, Nakano K, Antisense Oligonucleotides Targeting Y-Box Binding Protein-1 Inhibit Tumor Angiogenesis by Downregulating Bcl-xL-VEGFR2/-Tie Axes, Mol Ther Nucleic Acids, 10.1016/j.omtn, 9, 170-181, 2017.12, Y-box binding protein-1 (YB-1), involved in cancer progression and chemoradiation resistance, is overexpressed in not only cancer cells but also tumor blood vessels. In this study, we investigated the potential value of amido-bridged nucleic acid (AmNA)-modified antisense oligonucleotides (ASOs) targeting YB-1 (YB-1 ASOA) as an antiangiogenic cancer therapy. YB-1 ASOA was superior to natural DNA-based ASO or locked nucleic acid (LNA)-modified YB-1 ASO in both knockdown efficiency and safety, the latter assessed by liver function. YB-1 ASOA administered i.v. significantly inhibited YB-1 expression in CD31-positive angiogenic endothelial cells, but not in cancer cells, in the tumors. With regard to the mechanism of its antiangiogenic effects, YB-1 ASOA downregulated both Bcl-xL/VEGFR2 and Bcl-xL/Tie signal axes, which are key regulators of angiogenesis, and induced apoptosis in vascular endothelial cells. In the xenograft tumor model that had low sensitivity to anti-VEGF antibody, YB-1 ASOA significantly suppressed tumor growth; not only VEGFR2 but also Tie2 expression was decreased in tumor vessels. In conclusion, YB-1/Bcl-xL/VEGFR2 and YB-1/Bcl-xL/Tie signal axes play pivotal roles in tumor angiogenesis, and YB-1 ASOA may be feasible as an antiangiogenic therapy for solid tumors..
68. Hidaka N¹, Sato Y¹, Kido S¹, Fujita Y¹, Kato K¹, Expectant management of pregnancies complicated by fetal growth restriction without any evidence of placental dysfunction at term: Comparison with routine labor induction., The journal of obstetrics and gynaecology research, 10.1111/jog.13461, 44, 1, 93-101, 2017.09, AIM: To assess the feasibility and practicality of expectant management for pregnancies with fetal growth restriction (FGR) at term without evidence of placental dysfunction.
METHODS: We reviewed the records of pregnancies with an estimated fetal weight ≤ 1.5 SD below the mean at 37 weeks of gestation. We excluded elective cesarean deliveries and pregnancies that, at 37 weeks, were complicated by oligohydramnios, decreased fetal cerebroplacental ratio, or pregnancy-related hypertensive disorders. Prior to May 2013, we performed routine labor induction for FGR at term; after that time, we used routine expectant management. The rate of delivery by cesarean or instrumental assist and the rate of neonatal morbidity were compared between the groups.
RESULTS: The gestational age at delivery and the neonatal birthweight were higher in the expectant management policy group (39+4 vs 38+1 weeks; 2405 vs 2205 g). The cesarean rate (7/77 vs 7/73) and the instrumental delivery rate (5/77 vs 6/73) did not differ. Neonatal hypoglycemia and hyperbilirubinemia were significantly less frequent (10/77 vs 21/73; 7/77 vs 20/73) in the expectant management policy group. Seven patients in the expectant management policy group underwent emergency cesarean delivery; five of these (71%) had required labor induction because of progression to oligohydramnios.
CONCLUSIONS: Expectant management policy for FGR at term can reduce neonatal morbidity without increasing maternal risk or the cesarean rate. Caution should be used, however, during labor if oligohydramnios develops during expectant management..
69. Morokuma S, Michikawa T, Yamazaki S, Nitta H, Kato K, Association between exposure to air pollution during pregnancy and false posi-tives in fetal heart rate monitoring., Scientific reports, 10.1038/s41598-017-12663-2, 7, 1, 1-8, 2017.09, Fetal heart rate (FHR) monitoring is essential for fetal management during pregnancy and delivery but results in many false-positive diagnoses. Air pollution affects the uterine environment; thus, air pollution may change FHR reactivity. This study assessed the association between exposure to air pollution during pregnancy and FHR monitoring abnormalities using 2005-2010 data from the Japan Perinatal Registry Network database. Participants were 23,782 singleton pregnant women with FHR monitoring, without acidemia or fetal asphyxia. We assessed exposure to air pollutants, including particulate matter (PM), ozone, nitrogen dioxide (NO2), and sulfur dioxide (SO2). In a multi-trimester model, first-trimester PM exposure was associated with false positives in FHR monitoring (odds ratio [OR] per interquartile range (10.7 μg/m3) increase = 1.20; 95% CI: 1.05-1.37), but not second-trimester exposure (OR = 1.05; 95% CI: 0.91-1.21) and third-trimester exposure (OR = 1.06; 95% CI: 0.96-1.17). The association with first-trimester PM exposure persisted after adjustment for exposure to ozone, NO2, and SO2; however, ozone, NO2, and SO2 exposure was not associated with false positives in FHR monitoring. First-trimester PM exposure may alter fetal cardiac response and lead to false positives in FHR monitoring..
70. Kido S, Hidaka N, Sato Y, Fujita Y, Miyoshi K, Nagata K, Taguchi T, Kato K, Re-evaluation of lung to thorax transverse area ratio immediately before birth in predicting postnatal short-term outcomes of fetuses with isolated left-sided congenital diaphragmatic hernia: a single center analysis., Congenit Anom (Kyoto), 10.1111/cga.12243, 1-6, 2017.08, We aimed to investigate whether the lung-to-thorax transverse area ratio (LTR) immediately before birth is of diagnostic value for the prediction of postnatal short-term outcomes in cases of isolated left-sided congenital diaphragmatic hernia (CDH). We retrospectively reviewed the cases of fetal isolated left-sided CDH managed at our institution between April 2008 and July 2016. We divided the patients into two groups based on LTR immediately before birth, using a cut-off value of 0.08. We compared the proportions of subjects within the two groups who survived until discharge using Fisher's exact test. Further, using Spearman's rank correlation, we assessed whether LTR was correlated with length of stay, duration of mechanical ventilation, and supplemental oxygen. Twenty-nine subjects were included (five with LTR < 0.08, and 24 with LTR ≥ 0.08). The proportion of subjects surviving until discharge was 40% (2/5) for patients with LTR < 0.08, as compared with 96% (23/24) for those with LTR ≥ 0.08. LTR measured immediately before birth was negatively correlated with the postnatal length of stay (Spearman's rank correlation coefficient, rs = -0.486), and the duration of supplemental oxygen (rs = -0.537). Further, the duration of mechanical ventilation was longer in patients with a lower LTR value. LTR immediately before birth is useful for the prediction of postnatal short-term outcomes in fetuses with isolated left-sided CDH. In particular, patients with prenatal LTR value less than 0.08 are at increased risk of postnatal death..
71. Morokuma S, Shimokawa M, Kato K, Sanefuji M, Shibata E, Tsuji M, Senju A, Kawamoto T, Kusuhara K, Maternal sleep and small for gestational age infants in the Japan Environment and Children's Study: a cohort study., BMC Res Notes, 10.1186/s13104-017-2675-9, 10, 1, 394-394, 2017.08, OBJECTIVES: Small for gestational age infants have an increased risk of immediate complications, short-term morbidity and mortality, and long-term neurologic and metabolic disorders in adulthood. Previous research has shown that reduced sleep duration is a risk factor for SGA birth. However, only a few studies have evaluated maternal sleep as a risk factor for SGA birth. In the present study, we investigated the relationship between the amount and quality of mothers' sleep and infants' birth weight.

RESULTS: This cohort study (n = 8631) used data from the Japan Environment and Children's Study, an ongoing cohort study that began in January 2011. Data on sleep status (sleep duration and one indicator of sleep quality) and potential confounding factors were recorded. A log-binomial regression model was used to estimate the risk of small for gestational age birth, and the results were expressed as risk ratios and their respective 95% confidence interval. No significant results were observed for sleep duration or tiredness upon waking. Neither the amount nor the quality of mothers' sleep was associated with the risk of small for gestational age birth..
72. Okawa H, Morokuma S, Maehara K, Arata A, Ohmura Y, Horinouchi T, Konishi Y, Kato K, Eye movement activity in normal human fetuses between 24 and 39 weeks of gestation., PLos One, 10.1371/journal.pone.0178722, 12, 7, 1-12, 2017.07, Rapid eye movement (REM) sleep occurs throughout a relatively large proportion of early development, and normal REM activity appears to be required for healthy brain development. The eye movements (EMs) observed during REM sleep are the most distinctive characteristics of this state. EMs are used as an index of neurological function postnatally, but no specific indices of EM activity exist for fetuses. We aimed to identify and characterize EM activity, particularly EM bursts suggestive of REM periods, in fetuses with a gestational age between 24 and 39 weeks. This cross-sectional study included 84 normal singleton pregnancies. Fetal EMs were monitored using real-time ultrasonography for 60 min and recorded as videos. The videos were manually converted into a time series of EM events, which were then analyzed by piecewise linear regression for various EM characteristics, including EM density, EM burst density, density of EMs in EM bursts, and continuous EM burst time. Two critical points for EM density, EM burst density, and density of EMs in EM bursts were evident at gestation weeks 28-29 and 36-37. Overall EM activity in human fetuses increased until 28-29 weeks of gestation, then again from 36-37 to 38-39 weeks of gestation. These findings may be useful for creating indices of fetal neurological function for prognostic purposes..
73. Michikawa T, Morokuma S, Fukushima K, Kato K, Nitta H, Yamazaki S, Maternal exposure to air pollutants during the first trimester and foetal growth in Japanese term infants, Environ Pollut, 10.1016/j.envpol.2017.06.069, 230, 387-393, 2017.06, Evidence supporting an inverse association between maternal exposure to air pollutants and foetal growth has been accumulating. However, the findings from Asian populations are limited, and the question of critical windows of exposure remains unanswered. We examined whether maternal exposure to air pollutants, in particular exposure during the first trimester (an important period of placental development), was associated with foetal growth in Japanese term infants. From the Japan Perinatal Registry Network database, we received birth data for 29,177 term singleton births in western Japan (Kyushu-Okinawa Districts) between 2005 and 2010. Exposure was expressed in terms of average concentrations of air pollutants (ozone, suspended particulate matter, nitrogen dioxide, and sulphur dioxide), as measured at the nearest monitoring stations to the respective delivery hospitals of the pregnant women, during the entire pregnancy and each trimester. As proxy markers of foetal growth restriction, we used small for gestational age (SGA), and adverse birth weight (low birth weight in addition to SGA). For pollutant exposure during the entire pregnancy, we did not observe the association with SGA and adverse birth weight. In the single-trimester model for the first trimester, however, we found a positive association between ozone exposure, and SGA (odds ratio [OR] per 10 ppb increase = 1.07, 95% confidence interval [CI] = 1.01-1.12) and adverse birth weight (OR = 1.07; 95% CI = 1.01-1.14). This association persisted in the multi-trimester model, and no association for exposure during the second or third trimester was observed. Exposure to other pollutants during each trimester was not associated with these outcomes. In conclusion, maternal exposure to ozone during the first trimester was independently associated with an elevated risk of poor foetal growth..
74. Sonoda K, Yahata H, Okugawa K, Kaneki E, Nakatsuki K, Naka M, Terado N, Ookubo F, Oda Y, Tsunehisa Kaku, Kato K, Isthmic-vaginal cytological findings after a trachelectomy for early-stage cervical cancer. , Cytopathology, 10.1111/cyt.12379, 28, 3, 243-245, 2017.06.
75. Michikawa T, Morokuma S, Nitta H, Kato K, Yamazaki S, Comparison between air pollution concentrations measured at the nearest monitoring station to the delivery hospital and those measured at stations nearest the residential postal code regions of pregnant women in Fukuoka, Environmental Health and Preventive Medicine 2017 22:55, 10.1186/s12199-017-0663-2, 1-7, 2017.06, Background:Numerous earlier studies examining the association of air pollution with maternal and foetal health estimated maternal exposure to air pollutants based on the women’s residential addresses. However, residential addresses, which are personally identifiable information, are not always obtainable. Since a majority of pregnant women reside near their delivery hospitals, the concentrations of air pollutants at the respective delivery hospitals may be surrogate markers of pollutant exposure at home. We compared air pollutant concentrations measured at the nearest monitoring station to Kyushu University Hospital with those measured at the closest monitoring stations to the respective residential postal code regions of pregnant women in Fukuoka.

Methods:Aggregated postal code data for the home addresses of pregnant women who delivered at Kyushu University Hospital in 2014 was obtained from Kyushu University Hospital. For each of the study’s 695 women who resided in Fukuoka Prefecture, we assigned pollutant concentrations measured at the nearest monitoring station to Kyushu University Hospital and pollutant concentrations measured at the nearest monitoring station to their respective residential postal code regions.

Results:Among the 695 women, 584 (84.0%) resided in the proximity of the nearest monitoring station to hospital or one of the four other stations (as the nearest stations to their respective residential postal code region) in Fukuoka city. Pearson’s correlation for daily mean concentrations among the monitoring stations in Fukuoka city was strong for fine particulate matter (PM2.5), suspended particulate matter (SPM), and photochemical oxidants (Ox) (coefficients ≥0.9), but moderate for coarse particulate matter (the result of subtracting the PM2.5 from the SPM concentrations), nitrogen dioxide, and sulphur dioxide. Hospital-based and residence-based concentrations of PM2.5, SPM, and Ox were comparable.

Conclusions:For PM2.5, SPM, and Ox, exposure estimation based on the delivery hospital is likely to approximate that based on the home of pregnant women..
76. Wada T, Ohishi Y, Kaku T, Aman M, Imamura H, Yasutake N, Sonoda K, Kato K, Oda, Y, Endocervical Adenocarcinoma With Morphologic Features of Both Usual and Gastric Types: Clinicopathologic and Immunohistochemical Analyses and High-risk HPV Detection by In Situ Hybridization., Am J Surg Pathol, 10.1097/PAS.0000000000000833, 41, 5, 696-705, 2017.05, The fourth edition of the World Health Organization classification set up new entities of endocervical adenocarcinoma (ECA), namely the "usual type" and "gastric type." These 2 types are considered to be distinct histogenetically because of their differing immunophenotypes, human papillomavirus (HPV) status, and prognoses. Usual-type ECAs (U-ECAs) are virtually always associated with high-risk human papillomavirus (HR-HPV) infection. Gastric-type ECAs (G-ECAs) are believed not to be associated with HR-HPV infection. Morphologically, U-ECA cells are characterized by mucin-poor and eosinophilic cytoplasm, resembling endometrioid carcinoma (a pseudoendometrioid feature). G-ECA cells are characterized by abundant clear or pale, mucinous cytoplasm and distinct cell borders. However, in routine practice we noticed that some ECAs contain morphologically usual type-like components and gastric type-like components in a single tumor; we have named these "G+U" ECAs. The histogenesis of such tumors has not been investigated. We conducted the present study to clarify the clinicopathologic and immunohistochemical features and HPV status of G+U ECAs, and to determine whether G+U ECAs are genuine G-ECAs mimicking U-ECAs or genuine U-ECAs with gastric type-like morphology. We retrospectively analyzed a series of 70 consecutive cases of ECA diagnosed as mucinous ECA, endocervical type, and we reclassified them on the basis of the latest World Health Organization classification. We identified 48 (69%) pure U-ECAs, 9 pure G-ECAs, and 13 G+U ECAs. Ten of the 13 G+U ECAs (77%) showed no HR-HPV infection by in situ hybridization (HPV-unrelated G+U ECAs) and showed frequent HIK1083 expression and aberrant p53 expression in both usual type-like and gastric type-like components. The other 3 G+U ECAs showed HR-HPV infection (HPV-related G+U EACs) and frequent p16+/p53-/HIK1083- immunophenotype in both usual type-like and gastric type-like components. The U-ECAs were characterized by HR-HPV infection detected by in situ hybridization and frequent p16+/p53-/HIK1083- immunophenotype, similar to that of the HPV-related G+U ECAs. In contrast, the pure G-ECAs were characterized by the absence of HPV infection and frequent HIK1083 expression and aberrant p53 expression, similar to that of HPV-unrelated G+U ECAs. G+U ECAs thus represent a heterogenous group composed of genuine G-ECAs and genuine U-ECAs. Most of the G+U ECAs we examined were genuine HPV-unrelated G-ECAs with usual type-like components showing mucin-poor, eosinophilic cytoplasm (pseudoendometrioid morphology). A small population of G+U ECAs was genuine HPV-related U-ECAs with gastric type-like components showing mucin-rich, voluminous cytoplasm. Thus, both types of ECAs can occasionally display patterns of differentiation suggesting a component of the other type but true mixed tumors do not appear to exist. Ancillary techniques (immunohistochemical analysis of p16, p53, and HPV DNA detection assays) should be used to assure proper classification of tumors with mixed morphologic features..
77. Yumoto Y, Jwa SC, Wada S, Takahashi Y, Ishii K, Kato K, Usui N, Sago H, The outcomes and prognostic factors of fetal hydrothorax associated with trisomy 21., Prenatal Diagnosis, 10.1002/pd.5066, 37, 1-7, 2017.05, OBJECTIVES: To determine the characteristics, outcomes, and prognostic factors of fetal hydrothorax (FHT) with trisomy 21.
METHODS: A nationwide survey was conducted on FHT fetuses with trisomy 21 delivered after 22 weeks' gestation between January 2007 and December 2011 at perinatal centers.
RESULTS: The 91 cases of FHT with trisomy 21 included 28 (30.8%) diagnosed in utero and 63 (69.2%) diagnosed after birth. The natural remission rate was 6.6% (6/91). Thoracoamniotic shunting was performed in 14.3% (13/91) of cases. The survival rates of the hydropic, nonhydropic, and total cases were 47.0% (31/66), 84.0% (21/25), and 57.1% (52/91), respectively. The crude odds ratio for death was 8.2 (p = 0.003) for fetuses diagnosed at 26-30 weeks of gestational age (vs ≥30 weeks), 5.9 (p = 0.003) for hydrops, 4.0 (p = 0.04) for bilateral pleural effusion, 0.68 (p = 0.42) for associated cardiovascular anomalies, and 2.1 (p = 0.26) for thoracoamniotic shunting (vs no fetal therapy).
CONCLUSIONS: The prognosis of FHT with trisomy 21 was not very poor, but it was still worse than that of primary FHT. Hydrops, an early gestational age at the diagnosis and bilateral effusion, but not associated anomalies, were risk factors for death. Fetal therapy showed no survival benefit for FHT with trisomy 21..
78. Aman M, Ohishi Y, Imamura H, Shinozaki T, Yasutake N, Kato K, Oda Y, Expression of protease-activated receptor-2 (PAR-2) is related to advanced clinical stage and adverse
prognosis in ovarian clear cell carcinoma., Human Pathology, 10.1016/j.humpath.2017.04.008, 64, 156-163, 2017.04, Recent studies demonstrated that Protease-activated receptor-2 (PAR-2) correlates with tumor progression in various tissues. On the other hand, oxidative stress arising from endometriosis has been considered a cause of carcinogenesis in ovarian clear cell carcinoma (OCCC). We previously demonstrated that oxidative stress upregulates PAR-2 expression, and we conducted the present study to investigate the PAR-2 expression and its relation to clinicopathological factors and oxidative stress in OCCC. We performed an immunohistochemical evaluation in 95 cases of OCCC. For the evaluation of oxidative stress markers, 31 cases of ovarian endometrioid carcinoma (OEC) were also examined. No significant differences in the expression of COX2 and iNOS were observed between OCCC and OEC. Sixty-two percent of the OCCC cases showed high 8-OHdG expression, whereas all of the OEC cases showed almost negative immunoreactivities. The presence of endometriosis did not affect the expression of these oxidative stress markers or prognosis. High PAR-2 expression was observed in 20% (14/71) of the early FIGO stage cases but 58% (14/24) of the advanced FIGO stage cases. High PAR-2 expression was significantly correlated with advanced FIGO stage and shorter overall survival. We found no correlations between PAR-2 expression and oxidative stress in OCCC. Our results suggest that PAR-2 plays an important role in the progression of OCCC. The expression of 8-OHdG is a characteristic finding of OCCC, indicating that the injury of DNA by oxidative stress may be involved in the carcinogenesis of OCCC..
79. Okugawa K, Kobayashi H, Sonoda K, Kaneki E, Kawano Y, Hidaka N, Egashira K, Fujita Y, Yahata H, Kato K, Oncologic and obstetric outcomes and complications during pregnancy after fertility-sparing abdominal trachelectomy for cervical cancer:a retrospective review.
, Int J Oncol, 10.1007/s10147-016-1059-9, 22, 2, 340-346, 2017.04, BACKGROUND: Trachelectomy was developed as a fertility-sparing surgery for early-stage cervical cancer in patients of childbearing age. The purpose of this study is to evaluate oncologic and obstetric outcomes and complications after abdominal trachelectomy.

METHODS: We began to perform abdominal trachelectomy in 2005. Our institutional review board approved this clinical study, and fully informed consent was obtained from each patient. The medical records of patients who underwent trachelectomy were retrospectively reviewed.

RESULTS: We performed 151 abdominal trachelectomies (89 radical trachelectomies, 48 modified radical trachelectomies, and 14 simple trachelectomies). The median age of the patients was 33 years, and the median postoperative follow-up period was 61 months. Although one patient experienced recurrence at the preserved cervix, none died after treatment. A total of 61 patients attempted to conceive after trachelectomy, and 21 pregnancies were achieved in 15 women. Hence, the pregnancy rate among patients who attempted to conceive was 25%. Fifteen babies were delivered by cesarean section between gestational weeks 23 and 37. Six babies were delivered at term. Six cases of preterm premature rupture of the membranes occurred. Varices appeared around the uterovaginal anastomotic site in five patients.

CONCLUSIONS: Our data indicate that the oncologic outcome was excellent but infertility treatment was necessary to achieve the majority of conceptions. Additionally, preterm premature rupture of the membranes and premature delivery were frequently observed. An improved pregnancy rate and prevention of complications during pregnancy are issues that should be addressed in future studies.
80. Morokuma S, Tsukimori K, Hori T, Kato K, Furue M, The Vernix Caseosa is the Main Site of Dioxin Excretion in the Human Foetus., Sci Rep, 10.1038/s41598-017-00863-9, 7, 1, 739-739, 2017.04, Dioxins are highly toxic to foetuses and prenatal exposure leads to adverse health effects; however, the metabolic pathways involved in dioxin excretion are poorly understood. We determined the dynamics of maternal-to-foetal dioxin transfer during normal pregnancy and how foetuses eliminate polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and non-ortho polychlorinated biphenyls. Dioxin levels in maternal blood, cord blood, placenta, vernix caseosa, meconium, and amniotic fluid were analysed by high-resolution gas chromatography/mass spectrometry. The average levels of total dioxins, expressed as picograms of toxic equivalency quantity per gram of lipid and in parentheses, dioxin fraction, with maternal blood levels arbitrarily set as 100%, were as follows: maternal blood, 15.8 (100%); placenta, 12.9 (81.5%); cord blood, 5.9 (37.2%); vernix caseosa, 8.4 (53.2%); meconium, 2.9 (18.2%); and amniotic fluid, 1.5 (9.2%). Similar proportions were observed for each dioxin congener. Thus, the highest content of foetal dioxins was observed in the vernix caseosa, indicating that this is the major site of dioxin excretion in human foetuses..
81. Michikawa T, Seiichi M, Yamazaki S, Fukushima K, Kato K, Nitta H, Air Pollutant Exposure Within a Few Days of Delivery and Placental Abruption in Japan., Epidemiology, 10.1097/EDE.0000000000000605, 28, 2, 190-196, 2017.03, BACKGROUND: Placental abruption is an emergency obstetric complication. Although the etiology of abruption is not fully understood, acute stimuli, such as ischemia and/or inflammation, are associated with rupture of the decidual artery, resulting in placental separation. Ischemia and inflammation are acute biologic effects of air pollution. Using a case-crossover design, we tested the hypothesis that a short-term increase in exposure to air pollutants is a potential trigger of placental abruption.

METHODS: We received data for western Japan (Kyushu-Okinawa Districts) from the Japan Perinatal Registry Network database. From 2005 to 2010, 821 singleton pregnant women with placental abruption were identified. We assigned daily concentrations of air pollutants, including nitrogen dioxide (NO2), suspended particulate matter, ozone, and sulfur dioxide (SO2), from the nearest monitoring station to the respective delivery hospital of each woman. Because information on the onset day of abruption was not obtained, we assumed the case day to be 1 day before the day of delivery.

RESULTS: Exposure to NO2 at 2 days' lag was associated with placental abruption (temperature adjusted odds ratio per 10 ppb increase = 1.4; 95% confidence interval = 1.1, 1.8). The association patterns were similar, when we restricted to participants who delivered by emergency cesarean (1.4, 1.1, 1.9), or who delivered after 35 weeks of gestation (1.4, 1.0, 2.0). There was no association with suspended particulate matter, ozone, or SO2.

CONCLUSIONS: We observed an association between NO2 exposure at 2 days before the day of delivery and placental abruption in pregnant Japanese women..
82. Kitade S, Onoyama I, Kobayashi H, Yagi H, Kato M, Tsunematsu R, Asanoma K, Sonoda K, Wake N, Hata K, Nakayama K, Kato K, FBXW7 is involved in the acquisition of the malignant phenotype in epithelial ovarian Tumors., Cancer Sci, 10.1111/cas.13026., Epub ahead of print, 2016.08, FBXW7 is a ubiquitin ligase that mediates ubiquitylation of oncoproteins, such as c-Myc, cyclin E, Notch and
c-Jun. FBXW7 is a known tumor-suppressor gene, and mutations in FBXW7 have been reported in various
human malignancies. In this study, we examined the sequences of the FBXW7 and p53 genes in 57 ovarian
cancer clinical samples. Interestingly, we found no FBXW7 mutations associated with amino acid changes.
We also investigated FBXW7 expression levels in 126 epithelial ovarian tumors. FBXW7 expression was
negatively correlated with the malignant potential of ovarian tumors. That is to say, FBXW7 expression
levels in ovarian cancer samples were significantly lower than those in borderline and benign tumors
(P < 0.01). FBXW7 expression levels in serous carcinoma samples were the lowest among four major
histological subtypes. In addition, p53-mutated ovarian cancer samples showed significantly lower levels
of FBXW7 expression compared with p53 wild-type cancer samples (P < 0.001). DNA methylation arrays
and bisulfite PCR sequencing experiments revealed that 5'-upstream regions of FBXW7 gene in p53-mutated
samples were significantly higher methylated compared with those in p53 wild-type samples (P < 0.01).
This data indicates that p53 mutations might suppress FBXW7 expression through DNA hypermethylation
of FBXW7 5'-upstream regions. Thus, FBXW7 expression was downregulated in ovarian cancers, and was
associated with p53 mutations and the DNA methylation status of the 5'-upstream regions of FBXW7..
83. Morokuma S, Shimokawa M, Kato K, Sanefuji M, Shibata E, Tsuji M, Senju A, Kawamoto T, Kusuhara K, Relationship between hyperemesis gravidarum and small-for-gestational-age in the Japanese population:
the Japan Environment and Children's Study (JECS)., BMC pregnancy and childbirth, 10.1186/s12884-016-1041-6., 16, 247-247, 2016.08, BACKGROUND: Small-for-gestational-age in infancy is a known risk factor not only for short-term prognosis
but also for several long-term outcomes, such as neurological and metabolic disorders in adulthood.
Previous research has shown that severe nausea and vomiting in early pregnancy (NVP) and hyperemesis
gravidarum, which is an extreme form of NVP, represent risk factors for small-for-gestational-age birth.
However, there is no clear consensus on this association. Thus, in the present study, we investigated the
correlation between hyperemesis gravidarum and NVP on the one hand, and infant birth weight on the other,
using data from the Japan Environment and Children's Study (JECS).

METHODS: The data utilized in the present study were obtained from the JECS, an ongoing cohort study
that began in January 2011. Our sample size was 8635 parent-child pairs. The presence or absence of severe
NVP, hyperemesis gravidarum, and potential confounding factors were noted. A multivariable regression
analysis was used to estimate risks for small-for-gestational-age birth, and the results were expressed as risk
ratios and 95 % confidence intervals.

RESULTS: The risk ratios of small-for-gestational-age birth (95 % confidence interval) for mothers with
severe NVP and those with hyperemesis gravidarum were 0.86 (0.62-1.19) and 0.81 (0.39-1.66),
respectively, which represents a non-significant result.

CONCLUSIONS: In our analysis of JECS data, neither severe NVP nor hyperemesis gravidarum was associated with increased risk for small-for-gestational-age birth..
84. Yahata H, Kobayashi H, Sonoda K, Shimokawa M, Ogami T, Saito T, Ogawa S, Sakai K, Ichinoe A, Ueoka Y, Hasuo Y, Nishida M, Masuda S, Kato K, Efficacy of aprepitant for the prevention of chemotherapy-induced nausea and vomiting with a moderately
emetogenic chemotherapy regimen: a multicenter, placebo-controlled, double-blind, randomized study in
patients with gynecologic cancer receiving paclitaxel and carboplatin., Int J Clin Oncol, 10.1007/s10147-015-0928-y., 21, 3, 491-497, 2016.06, BACKGROUND: Substance P contributes to the hypersensitivity reaction (HSR) to paclitaxel in a rat model.
Aprepitant acts as an inhibitor of the binding of substance P to the neurokinin-1 receptor and, consequently,
may reduce the frequency of paclitaxel-induced HSR. While aprepitant has a prophylactic effect against
vomiting caused by high-dose cisplatin, the benefits of aprepitant have not been clearly demonstrated in
patients receiving paclitaxel and carboplatin (TC) combination chemotherapy.

METHODS: We conducted a multicenter, placebo-controlled, double-blind, randomized study in Japanese
patients with gynecologic cancer who received TC combination chemotherapy. Patients received aprepitant
or placebo together with both a 5-HT3 receptor antagonist and dexamethasone prior to chemotherapy.
The primary endpoint was the proportion of patients with HSR, and the secondary endpoints were the
proportion of patients with "no vomiting", "no significant nausea", and complete response, respectively.

RESULTS: Of the 324 randomized patients, 297 (151 in the aprepitant group; 146 in the placebo group)
were evaluated. The percentage of patients with HSR (9.2 vs. 7.5 %, respectively; P = 0.339) was not
significantly different between the groups. The percentage of "no vomiting" patients (78.2 vs. 54.8 %;
P < 0.0001), "no significant nausea" patients (85.4 vs. 74.7 %; P = 0.014), and patients showing complete
response (61.6 vs. 47.3 %, P = 0.0073) was significantly higher in the aprepitant group than in the placebo

CONCLUSION: The administration of aprepitant did not have a prophylactic effect on the HSR but was
effective in reducing nausea and vomiting in gynecologic cancer patients receiving TC combination
85. Michikawa T, Morokuma S, Yamazaki S, Fukushima K, Kato K, Nitta H, Exposure to air pollutants during the early weeks of pregnancy, and placenta praevia and pla-centa accreta in the western part of Japan., Environ Int, 10.1016/j.envint.2016.04.037, 92-93, 464-470, 2016.05, BACKGROUND: Placenta praevia is an obstetric complication involving placental implantation in the lower uterine segment. Given the suggested aetiology of placenta praevia, adverse biological effects of air pollutants, such as plasma viscosity increment, endothelial dysfunction, and systemic inflammation, have the potential to induce low implantation. We explored the association between exposure to air pollutants during the pregnancy period up to implantation, and placenta praevia, in pregnant Japanese women. The outcome also included placenta accreta, which often exists in combination with placenta praevia.

METHODS: From the Japan Perinatal Registry Network database, we obtained data on 40,573 singleton pregnant women in western Japan (Kyushu-Okinawa Districts) between 2005 and 2010. We assigned pollutant concentrations (suspended particulate matter [SPM], ozone, nitrogen dioxide [NO2], and sulphur dioxide [SO2]), measured at the nearest monitoring station to the respective delivery hospital of each woman. A logistic regression model was used to adjust for several covariates.

RESULTS: The odds ratios (ORs) of placenta praevia per 10 units increase were 1.12 (95% confidence interval (CI)=1.01-1.23) for SPM over 0-4weeks of gestation, and 1.08 (1.00-1.16) for ozone. The association between exposure to NO2 and SO2, and praevia, was in the direction of increased risk. SPM exposure during 0-4weeks was associated with placenta accreta without praevia (OR=1.33, 95% CI=1.07-1.66). We found no association with exposure to air pollutants during 5-12weeks and the second trimester.

CONCLUSIONS: Exposure to air pollutants through to implantation was positively associated with placenta praevia and accreta.
86. Masuda A, Katoh N, Nakabayashi K, Kato K, Sonoda K, Kitade M, Takeda S, Hata K, Tomikawa J, An improved method for isolation of epithelial and stromal cells from the human endometrium., The Journal of reproduction and development, 10.1262/jrd.2015-137, 62, 2, 213-218, 2016.04, We aimed to improve the efficiency of isolating endometrial epithelial and stromal cells (EMECs and EMSCs) from the human endometrium. We revealed by immunohistochemical staining that the large tissue fragments remaining after collagenase treatment, which are usually discarded after the first filtration in the conventional protocol, consisted of glandular epithelial and stromal cells. Therefore, we established protease treatment and cell suspension conditions to dissociate single cells from the tissue fragments and isolated epithelial (EPCAM-positive) and stromal (CD13-positive) cells by fluorescence-activated cell sorting. Four independent experiments showed that, on average, 1.2 × 10(6) of EMECs and 2.8 × 10(6) EMSCs were isolated from one hysterectomy specimen. We confirmed that the isolated cells presented transcriptomic features highly similar to those of epithelial and stromal cells obtained by the conventional method. Our improved protocol facilitates future studies to better understand the molecular mechanisms underlying the dynamic changes of the endometrium during the menstrual cycle..
87. Toomine Yukie, Sumiko Watanabe, Setsuo Sugishima, Yoshihiro Ohishi et al., Tamiya Sadafumi, Kobayashi Hiroaki, Sonoda Kenzo, Oda Yoshinao, Kato Kiyoko, Kaku Tsunehisa, Diagnostic value of squamous cell change associated with endometrial carcinoma: A cytopathologic approach., Diagn Cytopathol, 10.1002/dc.23418, 44, 3, 187-194, 2016.03, BACKGROUND: To determine the frequency of squamous cell change associated with endometrial carcinoma, to evaluate the relationship between squamous cell change and clinicopathological features, and to assess cytological findings for squamous cells with and without nuclear atypia in endometrial smears.

METHODS: In 103 cases of endometrioid adenocarcinoma of the endometrium having both histological and cytological specimens, the frequencies and relationships between the presence and absence of squamous cell changes were evaluated, as were the clinicopathological features of such changes. In endometrial smears, squamous cells with and without nuclear atypia were clinicopathologically assessed.

RESULTS: Squamous cell changes were found in 58.3% of cases that had both histological and cytological preparations. There were no significant differences between the group with squamous cell changes and the group without in any of the clinicopathological features. In the cytological smears, 70.0% of the 60 cases that showed squamous cell changes in both preparations did not have nuclear atypia of squamous cells, while 30.0% of those cases had atypia. The group of cases with squamous cells without atypia tended to be better differentiated than the group with atypia. Vessels were permeated significantly more often in the group with atypia than in the group without.

CONCLUSIONS: Our data suggest the importance of observing squamous cells in endometrial cytology, especially concerning findings on nuclei with vs without atypia, when endometrial carcinoma is suspected..
88. Yagi H, Asanoma K, Ohgami T, Ichinoe A, Sonoda K, Kato K, GEP oncogene promotes cell proliferation through YAP activation in ovarian cancer., Oncogene, 10.1038/onc.2015.505., 2016.01, G-protein-coupled receptors (GPCRs) and their ligands function in the progression of human malignancies. Gα12 and Gα13, encoded by GNA12 and GNA13, respectively, are referred to as the GEP oncogene and are implicated in tumor progression. However, the molecular mechanisms by which Gα12/13 activation promotes cancer progression are not fully elucidated. Here, we demonstrate elevated expression of Gα12/13 in human ovarian cancer tissues. Gα12/13 activation did not promote cellular migration in the ovarian cancer cell lines examined. Rather, Gα12/13 activation promoted cell growth. We used a synthetic biology approach using chimeric G proteins and GPCRs activated solely by artificial ligands to selectively trigger signaling pathways downstream of specific G proteins. We found that Gα12/13 promotes proliferation of ovarian cancer cells by activating the transcriptional coactivator YAP, a critical component of the Hippo signaling pathway. Furthermore, we reveal that inhibition of YAP by short hairpin RNA or a specific inhibitor prevented the growth of ovarian cancer cells. Therefore, YAP may be a suitable therapeutic target in ovarian cancer.Oncogene advance online publication, 25 January 2016; doi:10.1038/onc.2015.505..
89. Inagaki T, Kusunoki S, Tabu K, Okabe H, Yamada I, Taga T, Matsumoto A, Makino S, Takeda S, Kato K, Up-regulation of lymphocyte antigen 6 complex expression in side-population cells de-rived from a human trophoblast cell line HTR-8/SVneo., Human Cell, 10.1007/s13577-015-0121-7, 29, 1, 10-21, 2016.01, The continual proliferation and differentiation of trophoblasts are critical for the maintenance of pregnancy. It is well known that the tissue stem cells are associated with the development of tissues and pathologies. It has been demonstrated that side-population (SP) cells identified by fluorescence-activated cell sorting (FACS) are enriched with stem cells. The SP cells in HTR-8/SVneo cells derived from human primary trophoblast cells were isolated by FACS. HTR-8/SVneo-SP cell cultures generated both SP and non-SP (NSP) subpopulations. In contrast, NSP cell cultures produced NSP cells and failed to produce SP cells. These SP cells showed self-renewal capability by serial colony-forming assay. Microarray expression analysis using a set of HTR-8/SVneo-SP and -NSP cells revealed that SP cells overexpressed several stemness genes including caudal type homeobox2 (CDX2) and bone morphogenic proteins (BMPs), and lymphocyte antigen 6 complex locus D (LY6D) gene was the most highly up-regulated in HTR-8/SVneo-SP cells. LY6D gene reduced its expression in the course of a 7-day cultivation in differentiation medium. SP cells tended to reduce its fraction by treatment of LY6D siRNA indicating that LY6D had potential to maintain cell proliferation of HTR-8/SVneo-SP cells. On ontology analysis, epithelial-mesenchymal transition (EMT) pathway was involved in the up-regulated genes on microarray analysis. HTR-SVneo-SP cells showed enhanced migration. This is the first report that LY6D was important for the maintenance of HTR-8/SVneo-SP cells. EMT was associated with the phenotype of these SP cells..
90. Ohishi Y, Imamura H, Aman M, Shida K, Kaku T, Kato K, Oda Y, Is Invasive Micropapillary Serous Carcinoma a Low-grade Carcinoma?, Int J Gynecol Pathol, 10.1097/PGP.0000000000000211., 35, 1, 56-65, 2016.01, "Invasive micropapillary serous carcinoma" has been proposed as a synonym for low-grade serous carcinoma by some expert pathologists. In contrast, Singer and colleagues reported that some serous carcinomas with conspicuous invasive micropapillary pattern (SC-IMPs) can show high-grade nuclear atypia. However, the molecular features of such tumors have not been well documented. The aim of this study was to demonstrate and emphasize the fact that high-grade serous carcinoma confirmed by immunohistochemistry and molecular analysis can show conspicuous invasive micropapillary pattern. We selected 24 "SC-IMPs" and investigated: (1) their morphologic features; (2) the immunostaining pattern of p53 protein; and (3) KRAS/BRAF/TP53 gene mutations. The 24 SC-IMPs were subdivided into low-grade and high-grade tumors based primarily on the nuclear atypia, with the mitotic rate used as a secondary feature: low grade (n=5) and high grade (n=19). Low-grade SC-IMPs were characterized by low-mitotic activity, absence of abnormal mitosis, presence of serous borderline tumor, occasional BRAF mutation, and infrequent TP53 mutation. High-grade SC-IMPs were characterized by high-mitotic activity, presence of abnormal mitosis, conventional high-grade serous carcinoma, frequent TP53 mutation, and lack of KRAS/BRAF mutation. We demonstrated that high-grade serous carcinoma confirmed by aberrant p53 immunostaining and molecular analysis can show conspicuous invasive micropapillary pattern, validating Singer and colleague's report. Serous carcinoma with conspicuous invasive micropapillary pattern should not be readily regarded as low-grade serous carcinoma. Nuclear grade is the most important diagnostic feature in the SC-IMPs..
91. Miyata T, Sonoda K, Tomikawa J, Tayama C, Okamura K, Maehara K, Kobayashi H, Wake N, Kato K, Hata K, Nakabayashi K, Genomic, Epigenomic, and Transcriptomic Profiling towards Identifying Omics Features and Specific Biomarkers That Distinguish Uterine Leiomyosarcoma and Leiomyoma at Molecular Levels., Sarcoma 2015, 10.1155/2015/412068, 1-14, Epub 2015 Dec 28, 2015.12, Uterine leiomyosarcoma (LMS) is the worst malignancy among the gynecologic cancers. Uterine leiomyoma (LM),
a benign tumor of myometrial origin, is the most common among women of childbearing age. Because of their similar symptoms, it is difficult to preoperatively distinguish the two conditions only by ultrasound and pelvic MRI. While histopathological diagnosis is currently the main approach used to distinguish them postoperatively, unusual histologic variants of LM tend to be misdiagnosed as LMS. Therefore, development of molecular diagnosis as an alternative or confirmatory means will help to diagnose LMS more accurately. We adopted omics-based technologies to identify genome-wide features to distinguish LMS from LM and revealed that copy number, gene expression, and DNA methylation profiles successfully distinguished these tumors. LMS was found to possess features typically observed in malignant solid tumors, such as extensive chromosomal abnormalities, overexpression of cell cycle-related genes, hypomethylation spreading through large genomic regions, and frequent hypermethylation at the polycomb group target genes and protocadherin genes. We also identified candidate expression and DNA methylation markers, which will facilitate establishing postoperative molecular diagnostic tests based on conventional quantitative assays. Our results demonstrate the feasibility of establishing such tests and the possibility of developing preoperative and noninvasive methods..
92. Naka M, Ohishi Y, Kaku T, Watanabe S, Tamiya S, Ookubo F, Kato K, Oda Y, Identification of Intranuclear Inclusions is Useful for the Cytological Diagnosis of Ovarian Clear Cell Carcinoma, DIAGNOSTIC CYTOPATHOLOGY, 10.1002/dc.23322, 43, 11, 879-884, 2015.11, OBJECTIVE: The aim of this study was to clarify the diagnostic significance of the presence of intranuclear inclusions in clear cell carcinoma (CCC).

MATERIALS AND METHODS: We analyzed 98 imprint specimens and 53 ascites specimens from 98 ovarian carcinoma cases [28 CCCs, 37 serous carcinomas (SCs), 22 endometrioid carcinomas (ECs), and 11 mucinous carcinomas (MCs)]. We examined (1) frequency of intranuclear inclusion-positive cases of each ovarian carcinoma subtype, using imprint specimens, (2) frequency of intranuclear inclusion-positive cells of each ovarian carcinoma subtype, using imprint specimens, (3) frequency of intranuclear inclusion-positive cases of each ovarian carcinoma subtype, using ascites specimens, and (4) sensitivity and specificity of the presence of intranuclear inclusions for the cytological diagnosis of CCC.

RESULTS: (1) The frequency of intranuclear inclusion-positive cases in CCC (96.4%) was significantly higher than in SC (13.5%), EC (13.6%), and MC (18.2%) (P < 0.001). Two or more intranuclear inclusions in a single nucleus were observed only in CCC. (2) The frequency of intranuclear inclusion-positive cells in CCC (median, 0.41%) was significantly higher than in non-CCC subtypes (0.010%) (P < 0.001). (3) Using ascites specimens, the frequency of intranuclear inclusion-positive cases in CCC (78.6%) was significantly higher than in SC (10.3%), EC (0%), and MC (0%) (P < 0.001). (4) The sensitivity of intranuclear inclusions was 96.4%, and the specificity was 85.7%.

CONCLUSIONS: The identification of intranuclear inclusions, in particular a high frequency and multiple intranuclear inclusions in a single nucleus, is useful for the cytological diagnosis of CCC. Furthermore, these results may be applicable to ascites cytology. Diagn. Cytopathol. 2015;43:879-884. © 2015 Wiley Periodicals, Inc..
93. Matsueda S, Hidaka N, Kondo Y, Fujiwara A, Fukushima K, Kato K, External iliac artery thrombosis after common iliac artery balloon occlusion during cesarean hysterectomy for placenta accreta in cervico-isthmic pregnancy, JOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH, 10.1111/jog.12777, 41, 11, 1826-1830, 2015.11, Although the role of interventional radiology in the field of obstetrical hemorrhage has been widely reported upon recently, the rate of procedure-related complications has not been fully determined. We present the case of a patient who developed an external iliac artery thrombosis, a rarely reported complication associated with prophylactic common iliac artery balloon occlusion (CIABO). After CIABO, we found that the dorsalis pedis artery of the right foot was weak and the foot was cold, despite the fact that the patient had no complaints. Computed tomography demonstrated a linear thrombus in the right external iliac artery. We managed the patient conservatively using a heparin drip without the need for thromboembolectomy. Our experience suggests that it is important to consider the risk of thrombosis formation after CIABO. Physical examination post-procedure is key to identifying this complication early..
94. Takako Yamane, KAZUO ASANOMA, Hiroaki Kobayashi, Liu Ge, Hiroshi Yagi, Tatsuhiro Ogami, AKIMASA ICHINOE, Kenzo Sonoda, Norio Wake, Kiyoko Kato, Identification of the Critical Site of Calponin 1 for Suppression of Ovarian Cancer Properties., Anticancer Res., 35, 11, 5993-5999, 2015.11, BACKGROUND: Although several studies have demonstrated the tumor suppressive function of CNN1 (calponin 1), no studies have performed a site-specific analysis of CNN1 on tumor cell activities.

MATERIALS AND METHODS: We herein studied the site-specific effects of CNN1 in ovarian cancer cells using full-length CNN1 (fCNN1), three CNN1 repeats (3CNRs), or the first CNN1 repeat (CNR1) expression vectors. Ovarian cancer cells stably expressing each construct were analyzed for in vitro proliferation, cell motility, invasion, and soft agar assays. An in vitro model of pleural dissemination was also established.

RESULTS: Cell proliferation, anchorage-independent colony formation, cell motility, and cell invasion were all suppressed in fCNN1, 3CNRs, and CNR1-stably-expressing cells. CNN1 expression in mesothelial cells suppressed cancer cell invasion into a monolayer of mesothelial cells.

CONCLUSION: CNR1 showed similar suppressive effects as fCNN1. Results suggest CNR1 as a potential small synthetic peptide candidate for therapeutic strategies against ovarian cancer..
95. Imamura H, Ohishi Y, Aman M, Shida K, Shinozaki T, Yasutake N, Sonoda K, Kato K, Ovarian high-grade serous carcinoma with a noninvasive growth pattern simulating a serous borderline tumor, HUMAN PATHOLOGY, 10.1016/j.humpath.2015.06.002, 46, 10, 1455-1463, 2015.10, Ovarian serous borderline tumors (SBTs) being a precursor of low-grade serous carcinomas are morphologically characterized by noninvasive growth and low-grade cytology. On the other hand, many pathologists regard cytologically high-grade, noninvasive (HG-noninv) ovarian serous tumors resembling SBTs in low magnification as conventional high-grade serous carcinomas (HGSCs) by personal experiences. Nonetheless, there are no established molecular characteristic of such tumors. In this study, therefore, we attempted to provide the molecular evidence. We selected 37 ovarian serous tumors that exhibited a cytologically HG-noninv growth pattern, including 36 tumors that coexisted with conventional invasive HGSC components (HG-inv) and a single tumor exclusively composed of pure HG-noninv. Histologically, all HG-noninv showed many mitotic figures, and serous tubal intraepithelial carcinomas were identified in 3 tumors with HG-noninv. Immunohistochemically, most HG-noninv showed aberrant p53 expression, frequent IMP3 positivity, p16 overexpression, a high MIB-1 labeling index, and infrequent PAX2. By molecular analysis, the pure HG-noninv and 13 HGSCs with HG-noninv showed TP53 mutations, but KRAS/BRAF mutations were not detected in any of them. In 1 tumor, we detected an identical TP53 mutation in both HG-noninv and HG-inv components by using laser capture microdissection. These immunohistochemical and molecular features of HG-noninv were similar to those of conventional invasive HGSCs but different from those of SBTs. In conclusion, our results showed that a cytologically HG-noninv growth pattern simulating an SBT is a morphological spectrum of HGSC, but not a true SBT..
96. Michikawa T, Morokuma S, Fukushima K, Ueda K, Takeuchi A, Kato K, Nitta H, A register-based study of the association between air pollutants and hypertensive disorders in pregnancy among the Japanese population., Envionmental research, 10.1016/j.envres.2015.08.024., 142, 644-650, 2015.10, BACKGROUND:
Ambient air pollution is hypothesized to be a risk factor for hypertensive disorders in pregnancy, one of the major pregnancy complications. Past studies have reported the supporting evidence, however this mainly referred to the Western population, and results from trimester-specific analysis have been varied. In this study, we focused on exposure during the first trimester of pregnancy (placental development stage), and tested the hypothesis among the Japanese population.

We drew on data from the Japan Perinatal Registry Network database, and studied 36,620 singleton pregnant women without medical complications, in western Japan (Kyushu and Okinawa districts) between 2005 and 2010. In addition, data on ozone, suspended particulate matter (SPM), nitrogen dioxide (NO2), and sulfur dioxide (SO2) concentrations were obtained. The nearest monitoring station to the respective birthing hospital was used as a reference point for assigning average concentrations of each pollutant during the first trimester of pregnancy for each woman. The logistic regression model was applied to assess the association between quintiles of each pollutant and hypertensive disorders in pregnancy.

Mean concentrations during the first trimester were 41.3ppb for ozone, 27.4μg/m(3) for SPM, 11.8ppb for NO2, and 3.2ppb for SO2. High exposure to ozone was associated with an increased risk of hypertensive disorders in pregnancy (for highest quintile vs. lowest: odds ratio=1.20, 95% confidence interval=1.01-1.42). With regard to SPM, NO2 and SO2, we did not obtain the results with constant directionality.

Ozone exposure during early pregnancy may be a risk factor for hypertensive disorders in pregnancy..
97. Hachisuga M, Oki S, Kitajima K, Ikuta S, Sumi T, Kato K, Wake N, Meno C, Hyperglycemia impairs left-right axis formation and thereby disturbs heart morphogenesis in mouse embryos, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 10.1073/pnas.1504529112, 112, 38, E5300-E5307, 2015.09, Congenital heart defects with heterotaxia are associated with pregestational diabetes mellitus. To provide insight into the mechanisms underlying such diabetes-related heart defects, we examined the effects of high-glucose concentrations on formation of the left-right axis in mouse embryos. Expression of Pitx2, which plays a key role in left-right asymmetric morphogenesis and cardiac development, was lost in the left lateral plate mesoderm of embryos of diabetic dams. Embryos exposed to high-glucose concentrations in culture also failed to express Nodal and Pitx2 in the left lateral plate mesoderm. The distribution of phosphorylated Smad2 revealed that Nodal activity in the node was attenuated, accounting for the failure of left-right axis formation. Consistent with this notion, Notch signal-dependent expression of Nodal-related genes in the node was also down-regulated in association with a reduced level of Notch signaling, suggesting that high-glucose concentrations impede Notch signaling and thereby hinder establishment of the left-right axis required for heart morphogenesis..
98. KAZUO ASANOMA, Liu Ge, Takako Yamane, Yoko Miyanari, Tomoka Takao, Hiroshi Yagi, Tatsuhiro Ogami, AKIMASA ICHINOE, Kenzo Sonoda, Norio Wake, Kiyoko Kato, Regulation Mechanism of TWIST1 Transcription by BHLHE40 and BHLHE41 in Cancer Cells., Mol Cell Biol, 35, 24, 4096-4109, 2015.09, BHLHE40 and BHLHE41 are basic helix-loop-helix type transcription factors that play key roles in multiple cell behaviors. BHLHE40 and BHLHE41 (BHLHE40/41) were recently shown to be involved in epithelial-to-mesenchymal transition (EMT). However, the precise mechanism of EMT control by BHLHE40/41 remains unclear. In the present study, we demonstrated that BHLHE40/41 expression was controlled in a pathological stage-dependent manner in human endometrial cancer (HEC). Our in vitro assays showed that BHLHE40/41 suppressed tumor cell invasion. BHLHE40/41 also suppressed the transcription of the EMT effectors, SNAI1, SNAI2, and TWIST1. We identified the critical promoter regions of TWIST1 for its basal transcriptional activity. We elucidated that the transcription factor, SP1, was involved in the basal transcriptional activity of TWIST1 and that BHLHE40/41 competed with SP1 for DNA binding to regulate gene transcription. This study is the first to report the detailed functions of BHLHE40 and BHLHE41 in the suppression of EMT effectors in vitro. Our results suggest that BHLHE40/41 suppress tumor cell invasion by inhibiting EMT in tumor cells. We propose that BHLHE40/41 are promising markers to predict the aggressiveness of each HEC case, and that molecular targeting strategies involving BHLHE40/41 and SP1 may effectively regulate HEC progression..
99. Satoru Kyo, Kiyoko Kato, Endometrial Cancer Stem Cell as a Potential Therapeutic Target., Semin Reprod Med, 33, 5, 341-349, 2015.09, Adult stem cells have recently been identified in several types of mature tissue and it has been also suggested that stem-like cells exist in cancerous tissues. It is believed that many cancer stem cells (CSCs) upregulate the expression of drug transporters, allowing them to efficiently pump antitumor agents out of the cells. CSCs reside in a quiescent state, making them resistant to chemotherapeutic agents that target rapidly cycling cells. They are also endowed with a more invasive and metastatic phenotype. These results indicate the requirement to develop a new target treatment for CSCs. There are several methods for the identification of CSCs; for example, detection by CSC markers, such as CD133, CD44, CD117(c-kit), aldehyde dehydrogenase 1 (ALDH1), and isolation of side population (SP), which are identified based on their ability to remove intracellular Hoechst 33342, a fluorescent dye. Here, we review recent articles that show the presence of stem cells in endometrial cancer and introduce the results of our own recent studies using CD133 or CD117 positive cells and SP cells..
100. Ohmaru T, Fujita Y, Sugitani M, Shimokawa M, Fukushima K, Kato K, Placental elasticity evaluation using virtual touch tissue quantification during pregnancy, PLACENTA, 10.1016/j.placenta.2015.06.008, 36, 8, 915-920, 2015.08, INTRODUCTION: Virtual touch tissue quantification (VTTQ) has been developed to evaluate tissue elasticity. Our previous study using delivered placentas showed increased elasticity in fetal growth restriction (FGR). Therefore, we investigated changes in placental elasticity during pregnancy, including complicated pregnancies.

METHODS: Based on complications, 199 women were divided into 5 groups (normal, FGR, pregnancy induced hypertension (PIH), diabetes mellitus and collagen disease), and shear wave velocity (SWV) of the placenta, measured using VTTQ, was compared. A cross-sectional study was performed with the 143 normal cases to construct the reference range. The association between placental SWV and the expression ratio of collagen fibers in the placenta stained with Masson's trichrome was determined.

RESULTS:The SWV was safely measured for all participants. The correlation between SWV and gestational weeks was not significant. The mean ± SD SWVs in the normal, FGR, and PIH groups were 0.98 ± 0.21, 1.28 ± 0.39, and 1.60 ± 0.45 m/sec, respectively. The FGR and PIH groups had significantly higher SWVs than that of the normal group. SWV and the expression ratio of collagen fibers were significantly correlated.

DISCUSSION: Based on the present findings, changes in SWV during pregnancy were associated with placental fibrosis, and increased SWV in PIH and/or FGR cases might be influenced by infarction, ischemic changes, and inflammation, as well as fibrosis. In conclusion, the measurement of placental SWV is potentially useful to evaluate the condition of the placenta during pregnancy..
101. Kanako Okamoto, Ryosuke TSUNEMATSU, Tomoko Tahira, Kenzo Sonoda, KAZUO ASANOMA, Hiroshi Yagi, Tomoko Yoneda, Kenshi Hayashi, Norio Wake, Kiyoko Kato, SNP55, a new functional polymorphism of MDM2-P2 promoter, contributes to allele-specific expression of MDM2 in endometrial cancers, BMC MEDICAL GENETICS, 10.1186/s12881-015-0216-8, 16, 1, 67, 2015.08, BACKGROUND:
The functional single nucleotide polymorphism (SNP) in the MDM2 promoter region, SNP309, is known to be associated with various diseases, particularly cancer. Although many studies have been performed to demonstrate the mechanism of allele-specific expression (ASE) on SNP309, they have only utilized in vitro techniques. It is unknown whether ASE of MDM2 is ascribed solely to SNP309, in vivo.

We attempted to evaluate ASE of MDM2 in vivo using post-labeling followed by automated capillary electrophoresis under single-strand conformation polymorphism conditions. For measuring a quantitative difference, we utilized the SNPs on the exons of MDM2 as markers, the status of which was heterozygous in a large population. To address the cause of ASE beyond 20 %, we confirmed sequences of both MDM2-3'UTR and promoter regions. We assessed the SNP which might be the cause of ASE using biomolecular interaction analysis and luciferase assay.

ASE beyond 20 % was detected in endometrial cancers, but not in cancer-free endometria samples only when an SNP rs1690916 was used as a marker. We suspected that this ASE in endometrial cancer was caused by the sequence heterogeneity in the MDM2-P2 promoter, and found a new functional polymorphism, which we labelled SNP55. There was no difference between cancer-free endometria and endometrial cancer samples neither for SNP55 genotype frequencies nor allele frequencies, and so, SNP55 alone does not affect endometrial cancer risk. The SNP55 status affected the DNA binding affinity of transcription factor Sp1 and nuclear factor kappa-B (NFκB). Transcriptional activity of the P2 promoter containing SNP55C was suppressed by NFκB p50 homodimers, but that of SNP55T was not. Only ASE-positive endometrial cancer samples displayed nuclear localization of NFκB p50.

Our findings suggest that both the SNP55 status and the NFκB p50 activity are important in the transcriptional regulation of MDM2 in endometrial cancers..
102. Yuka Otera, Seiichi Morokuma, Kotaro Fukushima, Ai Anami, Yasuo Yumoto, Yushi Itoh, Masayuki Ochiai, KIMIAKI HASHIGUCHI, Norio Wake, Haruhiko Sago, Kiyoko Kato, Neurological outcomes in Chiari type II malformations and their correlation to morpho-logical findings and fetal heart rate patterns: a retrospective study., 10.1186/s13104-015-1014-2, 27, 8, 57-57, 2015.02, BACKGROUND: Correlations among Chiari type II malformation (CMII) morphological findings, the proportion of fetal heart rate patterns corresponding to the quiet phase (QP), and neurological outcomes have yet to be investigated.

FINDINGS: The correlations among the morphological findings (i.e., the degree of ventriculomegaly, myelomeningocele levels, and degree of cerebellar herniation), proportion of time spent in QP, and developmental quotients (DQs) were analyzed in 22 children. The proportion of time spent in QP was compared between children with poor neurological outcomes (n = 9) and those with good outcomes (n = 13). Pearson's correlations and the Mann-Whitney U-test were used to assess for statistical significance; P < 0.05 was considered statistically significant. No significant differences were observed between the DQs and morphological findings, but the DQs and the proportion of time spent in QP were significantly correlated (r = 0.287, P = 0.01). The proportion of time spent in QP was significantly different between children with poor outcomes and those with good outcomes (median, 11% [range, 0-32%] vs. 28% [range, 2-55%]; P = 0.006).

CONCLUSIONS: The proportion of fetal heart rate patterns corresponding to the QP might be a useful predictor of neurological outcomes in 2-year-old children with CMII..
103. Liu Ge, KAZUO ASANOMA, Tomoka Takao, Tsukimori Kiyomi, Uchi Hiroshi, Masutaka Furue, Kiyoko Kato, Norio Wake, Aryl hydrocarbon receptor SNP -130 C/T associates with dioxins susceptibility through regulating its receptor activity and downstream effectors including interleukin 24., Toxicology Letters, 232, 2, 384-392, 2015.01, Dioxins are persistent environmental pollutants that cause multiple adverse health effects in humans, mainly through binding to the ligand-activated transcription factor, aryl hydrocarbon receptor (AhR). Genetic variation in AhR may modulate the susceptibility to dioxins. In this study, we aimed to evaluate the effects of the single nucleotide polymorphism (SNP) -130 C/T in the AhR promoter on dioxin-inducible gene transcription, and to investigate interleukin-24 (IL-24) and interleukin-1β (IL-1β) as proxies for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure. Using primary human chorionic stromal cells, we found that cells with the TT genotype showed higher AhR mRNA and protein levels than did those of the CC genotype. Microarray was carried out to analyze the gene expression profiles of cells (CC and TT genotype) after exposing the cells to TCDD. Several genes associated with human disorders were more highly up-regulated in cells of the TT genotype. Higher up-regulation of IL-24 and IL-1β mRNA in cells with the TT genotype was observed. Furthermore, blood samples from 64 Yusho patients who were accidentally exposed to high concentrations of dioxins were analyzed for the genotype, dioxins concentrations and serum levels of IL-24 and IL-1β. We observed higher serum IL-24 levels and lower serum IL-1β levels in Yusho patients with the TT genotype than in those with the CC genotype. AhR SNP -130 C/T affects serum IL-24 and IL-1β levels, independently of serum dioxins concentrations in Yusho patients. Our observations demonstrate that SNP -130 C/T modulates AhR expression and expression levels of IL-24 and IL-1β, and suggest an association of AhR SNP -130 C/T with the susceptibility to dioxins. .
104. Kenzo Sonoda, Hideaki Yahata, AKIMASA ICHINOE, Koru Okugawa, Eisuke Kaneki, Yoshiaki Kawano, Hironori Kenjo, Tatsuhiro Ogami, Hiroshi Yagi, Ohga Saiji, Kaori Asai, Katsumasa Nakamura, Hiroshi Honda, Kiyoko Kato, Retrospective Analysis of Concurrent Chemoradiation with Triweekly Cisplatin plus 5-Fluorouracil Versus Weekly Cisplatin in Cervical Cancer., 35, 6, 3447-3454, 2015.01, BACKGROUND/AIM: Concurrent chemoradiation (CCRT) is the standard treatment for locally advanced cervical cancer. The purpose of the study was to compare the outcomes of triweekly cisplatin plus 5-fluorouracil and weekly cisplatin regimens.

PATIENTS AND METHODS: We retrospectively reviewed data from 91 patients with stage IB1-IVA cervical cancer.

RESULTS: Out of 91 patients, 48 received triweekly CCRT and 43 received weekly CCRT. For triweekly CCRT, patients received a median of two chemotherapy cycles and median total doses of cisplatin and 5-fluorouracil were 210 mg/body and 8,525 mg/body, respectively. For weekly CCRT, patients received a median of five chemotherapy cycles and the median total dose of cisplatin was 252 mg/body. No statistically significant differences in overall survival or progression-free survival were noted between the two groups.

CONCLUSION: Both triweekly CCRT and weekly CCRT appear to have similar efficacy for cervical cancer patients, but the toxicities were better tolerable in weekly CCRT.
105. Kohei Miyata, Tomoko Miyata, Kazuhiko Nakabayashi, Kohji Okamura, Masashi Naito, Tomoko Kawai, Shuji Takada, Kiyoko Kato, Shingo Miyamoto, Kenichiro Hata, Hiroshi Asahara, DNA methylation analysis of human myoblasts during in vitro myogenic differentiation: de novo methylation of promoters of
muscle-related genes and its involvement in transcriptional down-regulation.
, 10.1093/hmg/ddu457, 24, 2, 410-423, 2015.01, Although DNA methylation is considered to play an important role during myogenic differentiation, chronological alterations in DNA methylation and gene expression patterns in this process have been poorly understood. Using the Infinium HumanMethylation450 BeadChip array, we obtained a chronological profile of the genome-wide DNA methylation status in a human myoblast differentiation model, where myoblasts were cultured in low-serum medium to stimulate myogenic differentiation. As the differentiation of the myoblasts proceeded, their global DNA methylation level increased and their methylation patterns became more distinct from those of mesenchymal stem cells. Gene ontology analysis revealed that genes whose promoter region was hypermethylated upon myoblast differentiation were highly significantly enriched with muscle-related terms such as 'muscle contraction' and 'muscle system process'. Sequence motif analysis identified 8-bp motifs somewhat similar to the binding motifs of ID4 and ZNF238 to be most significantly enriched in hypermethylated promoter regions. ID4 and ZNF238 have been shown to be critical transcriptional regulators of muscle-related genes during myogenic differentiation. An integrated analysis of DNA methylation and gene expression profiles revealed that de novo DNA methylation of non-CpG island (CGI) promoters was more often associated with transcriptional down-regulation than that of CGI promoters. These results strongly suggest the existence of an epigenetic mechanism in which DNA methylation modulates the functions of key transcriptional factors to coordinately regulate muscle-related genes during myogenic differentiation. .
106. Kazuhisa Hachisuga, Nobuhiro Hidaka, Yasuyuki Fujita, Kotaro Fukushima, Kiyoko Kato, Can we predict neonatal thrombocytopenia in offspring of women with idiopathic thrombocytopenic purpura?, Blood Res, 49(4): 259-264, 2014.12, BACKGROUND: We aimed to investigate which factors in the clinical profile of mothers with idiopathic thrombocytopenic purpura (ITP) can predict neonatal risk of thrombocytopenia.
METHODS: Data was retrospectively collected from all pregnant women with ITP who presented to our institution between 2001 and 2013. Neonatal offspring of these women were classified into 2 groups based on the presence or absence of neonatal thrombocytopenia (platelet count <100×10(9)/L). Several parameters were compared between the 2 groups, including maternal age, maternal platelet count, maternal treatment history, and thrombocytopenia in siblings. We further examined the correlation between maternal platelet count at the time of delivery and neonatal platelet count at birth; we also examined the correlation between the minimum platelet counts of other children born to multiparous women.
RESULTS: Sixty-six neonates from 49 mothers were enrolled in the study. Thrombocytopenia was observed in 13 (19.7%) neonates. Maternal treatment for ITP such as splenectomy did not correlate with a risk of neonatal thrombocytopenia. Sibling thrombocytopenia was more frequently observed in neonates with thrombocytopenia than in those without (7/13 vs. 4/53, P<0.01). No association was observed between maternal and neonatal platelet counts. However, the nadir neonatal platelet counts of first- and second-born siblings were highly correlated (r=0.87).
CONCLUSION: Thrombocytopenia in neonates of women with ITP cannot be predicted by maternal treatment history or platelet count. However, the presence of an older sibling with neonatal thrombocytopenia is a reliable risk factor for neonatal thrombocytopenia in subsequent pregnancies.
107. Kenzo Sonoda, Kiyoko Kato, A disintegrin and metalloproteinase 9 is involved in ectodomain shedding of receptor-binding cancer antigen expressed on Siso cells.
, Biomed Res Int, 10.1155/2014/482396, 1-13, 2014.10, In several human malignancies, the expression of receptor-binding cancer antigen expressed on SiSo cells (RCAS1) is associated with aggressive characteristics and poor overall survival. RCAS1 alters the tumor microenvironment by inducing peripheral lymphocyte apoptosis and angiogenesis, while reducing the vimentin-positive cell population. Although proteolytic processing, referred to as "ectodomain shedding," is pivotal for induction of apoptosis by RCAS1, the proteases involved in RCAS1-dependent shedding remain unclear. Here we investigated proteases involved in RCAS1 shedding and the association between tumor protease expression and serum RCAS1 concentration in uterine cancer patients. A disintegrin and metalloproteinase (ADAM) 9 was shown to be involved in the ectodomain shedding of RCAS1. Given the significant correlation between tumor ADAM9 expression and serum RCAS1 concentration in both cervical and endometrial cancer as well as the role for ADAM9 in RCAS1 shedding, further exploration of the regulatory mechanisms by which ADAM9 converts membrane-anchored RCAS1 into its soluble form should aid the development of novel RCAS1-targeting therapeutic strategies to treat human malignancies. .
108. Hitomi Okabe, Shintaro Makino, Kiyoko Kato, Kikumi Matsuoka, Hioyuki Seki, Satoru Takeda, The effect of progesterone on genes involved in preterm labor. , J Reprod Immunol, 10.1016/j.jri.2014.03.008, 104-105, 80-91, 2014.10, The decidua is known to be a major source of intrauterine PGF2α during late gestation and labor, and inflammatory cytokines, including IL-1β, IL-6, and IL-8, are elevated in spontaneous preterm deliveries. In the present study, to elucidate how progesterone blocks the pathways associated with preterm birth, we determined the effects of P4 on the expression of PTGS-2 and PTGFR mRNA in human decidua fibroblast cells, as well as the genes, using microarray analysis. Senescence was induced in primary cultured human decidual cells treated with IL-1β. The IL-1β treatment implicated by microarray analysis increased gene expression levels of PTGS-2, PTGFR, NFκ-B p65, IL-17, and IL-8. In contrast, P4+IL-1β decreased the expression levels of all of these genes in comparison to treatment with IL-1β alone (p<0.05). IL-1β also increased the proportion of SA-β-gal-positive cells. Treatment with IL-1β also increased the p21 protein level in comparison to cells treated either with the vehicle or P4. Neither the p21 protein level nor the number of SA-β-gal-positive cells was increased in normal endometrial glandular cells by IL-1β (p<0.05). Our studies demonstrated that P4 changes the level of gene expression in a manner that favors an anti-inflammatory milieu. Because IL-8 appears to be the cytokine whose expression is most significantly modulated by P4, further studies evaluating IL-8 as a therapeutic target are needed. .
109. Nurismangul Yusuf, Tetsunori Inagaki, Soshi Kusunoki, Hitomi Okabe, Izumi Yamada, Akemi Matsumoto, Yasuhisa Terao, Satoru Takeda, Kiyoko Kato, SPARC was overexpressed in human endometrial cancer stem-like cells and promoted migration activity.
, Gynecologic Oncology, 134, 2, 356-363, 2014.08, Objectives
We previously demonstrated that side-population (SP) cells found in human endometrial cancer tissue have features of cancer stem cells (CSCs). Endometrial cancer SP cells show enhanced migration, the potential to differentiate into the mesenchymal cell lineage, and they are associated with the epithelial–mesenchymal transition (EMT). In this study, we analyzed the expression and function of a specific protein, SPARC (secreted protein acidic and rich in cysteine) which we found to be up-regulated in endometrial cancer.

We performed microarray expression analysis to screen for up-regulated genes in CSCs using a set of RK12V-SP cells and -non-SP (NSP) cells. We used the MetaCore package to identify the Gene GO pathway MAPs associated with the up-regulated genes. Here, we investigated the expression and functions of SPARC, one of the genes up-regulated in endometrial CSCs.

We established SPARC-overexpressing cells by transfecting endometrial cancer cells (Ishikawa cells [IK-SPARC cells]). We characterized these cells' growth rate, tumorigenicity, migration and invasion activity. The levels and locations of SPARC protein expression in Hec1SP cells-derived tumors and endometrial cancer tissues were examined by immunohistochemistry.

SPARC was detected by microarray expression analysis during screens for up-regulated genes in SP and NSP CSC. The level of SPARC expression was enhanced in Hec1 SP cells compared with that in Hec1 non-SP cells. SPARC enhanced fibronectin expression and promoted migration activity in IK cells. SPARC expression suppressed tumor growth but promoted formation of tumor stroma.

SPARC was expressed in endometrial cancer tissues, in particular, poorly differentiated endometrioid adenocarcinoma, clear and serous adenocarcinoma,but not in normal endometrial tissue.

This is the first report of overexpression of SPARC in endometrial cancer stem-like cells. SPARC expression is associated with cell migration and stroma formation.

SPARC; Endometrial cancer; EMT; CSCs; Cell migration
110. Arisa Fujiwara, Kotaro Fukushima, Hirosuke Inoue, Takeshi Takashima, Hiromasa Nakahara, Shoji Satoh, Masayuki Ochiai, Toshiro Hara, Mototsugu Shimokawa, Kiyoko Kato, Perinatal management of preterm premature ruptured membranes affects neonatal prognosis.
, Journal of Perinatal Medicine, Volume 0, Issue 0, 1-7, 2014.01, Abstract Aim: To determine the factors affecting neonatal prognosis in preterm premature rupture of membranes (PPROM). Method: We conducted a case-control study involving 92 women between the years 2000 and 2010 diagnosed with PPROM between 25 and 31 weeks' gestation, who received antenatal steroids, and delivered between 26 and 31 weeks' gestation; a retrospective cohort study was conducted based on the results. We used data from four tertiary centers and compared the frequencies of neonatal neurologic deficits and neonatal deaths. Results: There was a difference between the two groups; specifically, the ND group (n=18) consisted of patients whose infants had neurologic deficits and/or neonatal deaths and the neurologically normal (NN) group (n=74) included NN neonates amongst the patients who had expectant management (94% vs. 73%, respectively). Multivariable analysis revealed that expectant management was independently associated with an increased risk for neonatal neurologic deficits and neonatal deaths (odds ratio, 16.14). All neonates with poor prognosis in the expectant-management group delivered within 14 days after PPROM. Conclusions: Expectant management within 14 days after PPROM is associated with poor neonatal outcomes. Decisions regarding an expectant strategy should be made carefully. An immediate, planned delivery after steroid administration should be considered to improve neonatal prognosis in patients who have PPROM after 26 weeks' gestation..
111. Masayuki Ochiai, Kinjo Tadamune, Yasushi Takahata, Mariko Iwayama, Takeru Abe, KENJI IHARA, Shouichi Ohga, Kotaro Fukushima, Kiyoko Kato, tomoaki taguchi, Toshiro Hara, Survival and Neurodevelopmental Outcome of Preterm Infants Born at 22-24 Weeks of Gestational Age, NEONATOLOGY, 10.1159/000355818, 105, 2, 79-84, 2013.11.
112. Seiichi Morokuma, Kotaro Fukushima, Kiyoko Kato, Relationship between arousal response in newborn infants before micturition and bed-wetting from 5 to 7 years of age, EARLY HUMAN DEVELOPMENT, 10.1016/j.earlhumdev.2013.08.020, 89, 12, 989-991, 2013.12.
113. Yuka Otera, Seiichi Morokuma, Kotaro Fukushima, Norio Wake, Kiyoko Kato, Correlation between regular mouthing movements and heart rate patterns during non-rapid eye movement periods in normal human fetuses between 32 and 40 weeks of gestation, EARLY HUMAN DEVELOPMENT, 10.1016/j.earlhumdev.2012.12.007, 89, 6, 381-386, 2013.06.
114. Maiko Sugitani, Yasuyuki Fujita, Kotaro Fukushima, Kiyoko Kato, A new method for measurement of placental elasticity: acoustic radiation force impulse imaging, Placenta, 34, 11, 1009-1013, 2013.11.
115. Soshi Kusunoki, Kiyoko Kato, Kouichi Tabu, Tetsunori Inagaki, Hitomi Okabe, Hiroshi Kaneda, Shin Suga, Yasuhisa Terao, Tetsuya Taga, Satoru Takeda, The inhibitory effect of salinomycin on the proliferation, migration and invasion of human endometrial cancer stem-like cells, GYNECOLOGIC ONCOLOGY, 10.1016/j.ygyno.2013.03.005, 129, 3, 598-605, 2013.06, Goals: We previously demonstrated that side-population (SP) cells in human endometrial cancer cells (Hec1 cells) and in rat endometrial cells expressing oncogenic human K-Ras protein (RK12V cells) have features of cancer stem cells (CSCs). Hec1-SP cells showed enhanced migration and
the potential to differentiate into the mesenchymal cell lineage. In this study, we analyzed the association of the epithelial-mesenchymal transition (EMT) with the properties of these endometrial CSCs. We also assessed and the effects of salinomycin (a compound with EMT-specific toxicity) on the proliferative capacity, migration and invasiveness of these endometrial CSCs using Hec1-SP cells.
Method: We performed microarray expression analysis to screen for up-regulated genes in CSCs using a set of RK12V-SP cells and –non-SP(NSP) cells and used the Metacore package to identify the Gene GO pathway MAPs involved in the up-regulated genes. To analyze their association with EMT, the expression of several EMT associated genes in Hec1-SP cells was investigated by real time PCR and compared with that in Hec1-NSP cells. We assessed the expression of BAX, BCL2, LEF1, cyclinD and fibronectin by real time PCR. We also evaluated the viabilities, migration and invasive activities, and tumorigenicities of these SP cells and NSP cells in the presence or absence of salinomycin.
Results: We demonstrated that i) EMT processes were observed in both RK12V-SP cells and Hec1-SP cells, ii) the level of fibronectin was enhanced in Hec1-SP cells and salinomycin reduced the level of fibronectin expression, iii) salinomycin induced apoptosis and inhibited Wnt signaling, and iv) salinomycin inhibited the proliferation, migration, invasiveness and tumorigenicity of these SP cells.
Conclusion: This is the first report of an inhibitory effect of salinomycin on the properties of endometrial CSCs..
116. Tadahisa Takeuchi, Yoshihiro Ohishi, Hiroko Imamura, Murasaki Aman, Kaai Shida, Hiroaki Kobayashi, Kiyoko Kato, Yoshinao Oda, Ovarian Transitional Cell Carcinoma Represents a Poorly Differentiated Form of High-grade Serous or Endometrioid Adenocarcinoma, AMERICAN JOURNAL OF SURGICAL PATHOLOGY, 37, 7, 1091-1099, 2013.07.
117. Tomoko Yoneda, Ayumi Kuboyama, Kiyoko Kato, Tatsuhiro Ogami, Kanako Okamoto, Toshiaki Saito, Norio Wake, Association of MDM2 SNP309 and TP53 Arg72Pro polymorphisms with risk of endometrial cancer, ONCOLOGY REPORTS, 10.3892/or.2013.2433, 30, 1, 25-34, 2013.07.
118. Takao T, Asanoma K, Tsunematsu R, Kato K, Wake N, The maternally expressed gene Tssc3 regulates the expression of Mash2 transcription factor in mouse trophoblast stem cells through thr Akt-Sp1 signaling pathway, J Biol Chem.287(51):42685-42694,2012, 2012.12.
119. Takao T, Asanoma K, Tsunematsu R, Kato K, Wake N, The maternally expressed gene Tssc3 regulates the expression of Mash2 transcription factor in mouse trophoblast stem cells through thr Akt-Sp1 signaling pathway, J Biol Chem.287(51):42685-42694,2012, 2012.12.
120. Kiyoko Kato, Kusunoki S, Inagaki T, Yusuf N, Suga S, Terao Y, Arima T, Tsukimori K, Takeda S, Side-population cells derived from non-tumorigenic rat endometrial cells are a candidate cell of origin for malignant endometrial tumors, J Stem Cell Res Ther. doi:10.4172/2157-7633.S7-003, 2012.
121. Fukushima K, Tsukimori K, Li D, takao T, Morokuma S, Kato K, Seki H, Takeda S, matsumura S, Wake N, Effect of transient TCDD exposure on immortalized human trophoblast-derived cell lines., Hum Exp Toxicol. 31(6):550-556, 2012, 2012.05.
122. Takao T, Asanoma K, Kato K, Fukushima K, Tsunematsu R, Hirakawa T, Matsumura S, Seki H, Takeda S, Wake N: ,
Isolation and Characterization of Human Trophoblast Side-Population (SP) Cells in Primary Villous Cytotrophoblasts and HTR-8/SVneo Cell Line.

, PLoS One. , 6, 7, e21990, 2011.07.
123. Onoda A, Ueno T, Uchiyama S, Hayashi SI, Kato K, Wake N: ,
Effects of S-equol and natural S-equol supplement (SE5-OH) on the growth of MCF-7 in vitro and as tumors implanted into ovariectomized athymic mice.
, Food Chem Toxicol. , 49, 9, 2279-2284, 2011.09.
124. Kato K, Kuhara A, Yoneda T, Inoue T, Takao T, Ohgami T, Dan L, Kuboyama A, Kusunoki S, Takeda S, Wake N: , Sodium Butyrate inhibitis the Self-Renewal Capacity of Endometrial Tumor Side-Population Cells by Induction a DNA Damage Response.
, Mol Cancer Ther. , 10, 8, 1-10, 2011.09.
125. Kukita Y, Yahara K, Tahira T, Higasa K, Sonoda M, Yamamoto K, Kato K, Wake N, Hayashi K: ,
A definitive haplotype map as determined by genotyping duplicated haploid genomes finds a predominant haplotype preference at copy number variation events.
, Am J Hum Genet. , 86, 6, 918-928, 2009.06.
126. Ohgami T, Kato K, Kobayashi H, Sonoda K, Inoue T, Yamaguchi S, Yoneda T, Wake N: ,
Low-dose mithramycin exerts its anti-cancer effect via the p53 signaling pathway and synergizes with nutlin-3 in gynecologic cancers.

, Cancer Sci. , 101, 6, 1387-1395, 2010.06.
127. Higuchi M, Yamayoshi A, Kato K, Kobori A, Wake N, Murakami A:,
Specific regulation of point-mutated K-ras-immortalized cell proliferation by a photodynamic antisense strategy.
, Oligonucleotides. , 20, 1, 37-44, 2010.01.
128. Kato K, Takao T, Kuboyama A, Tanaka Y, Ohgami T, Yamaguchi S, Adachi S, Yoneda T, Ueoka Y, Kato K, Hayashi S, Asanoma K, Wake N: , Endometrial cancer side-population cells show prominent migration and have a potential to differentiate into the mesenchymal cell lineage. , Am J Pathol., 176, 1, 381-392, 2010.01.
129. Inoue T, Kato K, Kato H, Asanoma K, Kuboyama A, Ueoka Y, Yamaguchi S, Ohgami T, Wake N, Level of reactive oxygen species induced by p21Waf1/CIP1 is critical for the determination of cell fate. , Cancer Sci., 100, 7, 1275-1283, 2009.07.
130. Tanaka Y, Wake N, Kato K, Letter to Editor, Menopause, in press, 2009.05.
131. Fukushima K, Murata M, Hachisuga M, Tsukimori K, Seki H, Takeda S, Kato K, Wake N, Gene expression profiles by microarray analysis during matrigel-induced tube formation in a
human extravillous trophoblast cell line: comparison with endothelial cells
, Placenta, 29(10):898-904, 2008.10.
132. Higasa K, Kukita Y, Kato K, Wake N, Tahira T, Hayashi K, Evaluation of haplotype inference using definitive haplotype data obtained from complete
hydatidiform moles, and its significance for the analyses of positively selected regions
, PLoS Genetics, in press, 2009.05.
133. Inoue T, Kato K, Kato H, Asanoma K, Kuboyama A, Oogami T, Ueoka Y, Wake N, The level of reactive oxygen species induced by p21 WAF1/CIP1 is critical the determination of cell
, Cancer Sci, in press, 2009.05.
134. Yamaguchi S, Asanoma K, Takao T, Kato K, WakeN, Homeobox gene NECC1 is epigenetically
silenced in human uterine endometrial cancer and suppresses edtrogen-stimulated proliferation of
cancer cells by inhibiting serum response factor
, Int J Cancer, 124(11),2577-2588, 2009.01.
135. Matsushita I, Uchida S, Yamaguchi A, Tanaka Y, Ogawa S, Kato K, Tsukimori K, Wake N, Interstitial pregnancy in a woman with congenital afibrinogenemia, J Obstet Gynaecol Res, 34(5), 914-918, 2008.10.
136. Tanaka Y , Kato K , Mibu R , Uchida S , Asanoma K , Hashimoto K , Nozaki M , Wake N, Medroxyprogesterone acetate inhibits proliferation of colon cancer cell lines by modulating cell
cycle-related protein expression
, Menopause, 15:442-453, 2008.05.
137. Asanoma K,Kato H,Yamaguchi S,Shin CH,Liu ZP,Kato K,Inoue T,Miyanari Y,Yoshikawa K,Sonoda K,Fukushima K,Wake N, HOP/NECC1,a novel regulator of mouse trophoblast differentiation , J Biol Chem, 17:24065-74, 2007.08.
138. Yamayoshi A, Kato K, Suga S, Ichinose A, Arima T, Matsuda T, Kato H, Murakami A, Wake N. , Specific apotosis induction in HPV-positive cervical carcinoma cells by photodynamic antisense regulation, oligonucleotides, 17:66-79, 2007.05.
139. Kato K, Yoshimoto M, Kato K, Adachi S, Yamayoshi A, Arima T, Asanoma K, Kyo S, Nakahata T, Wake N, Characterization of side population cells (SP cells) in human normal endometrial cells
, Human Reproduction, 22:1214-23, 2007.04.
140. Suga S, Kato K, Yamayoshi A, Adachi S, Asanoma K, Yamaguchi S, Arima T, Kinoshita K, Wake N, An inhibitory effect on cell proliferation by blockage of the MAPK / Estrogen Receptor / MDM2 signal pathway in gynecologic cancer, Gynecologic Oncology, 105:341-350, 2007.03.
141. Arima T,Yamasaki K,John RM,Kato K,Sakumi K,Nakabeppu Y,Wake N,Kono T, The Human HYMAI/PLAGL1 differentially methylated region acts as an imprint control region in mice., Genomics, 88(5):650-8, 2006.08.
142. Ogura T, Kobayashi H, Ueoka Y, Okurgawa K, Kato K, Hirakawa T, Hashimoto S,Taniguchi S, Wake N, Nakano H , Adenovirus-mediated calponin h1 gene therapy directed against peritoneal dissemination of ovarian cancer : bifunctional therapeutic effects on peritoneal cell layer and cancer cells
, Clin Cancer Res, 12.5216-23, 2006.12.
143. Kato HD,Kondoh H,Inoue T,Asanoma K,Matsuda T,Arima T,Kato K,Yoshikawa T,Wake N, Expression of DCC and netrin-1 in normal human endometrium and its implication in endometrial carcinogenesis., GynecolOncol, 10.1016/j.ygyno.2004.07.050, 95, 2, 281-289, 95(2),281-9, 2004.11.
144. Arima T, Kato K, Hayashida T, Kamikihara T, Matsuda T, Kato H, Shirayoshi Y, Oshimura M, Soejima H, Mukai T, Wake N, Zac, LIT1 (KCNQ10T1) and p57KIP2 (CDKN1C) are in an imprinted gene network which may play a role in Beckwith-Wiedemann Syndrome., Nucleic Acids Research, 10.1093/nar/gki555, 33, 8, 2650-2660, 33, 8,2650-2660, 2005.01.
145. Ninomiya Y, Kato K, Takahashi A, Ueoka K, Kamikihara T, Arima T, Matsuda T,Kato H, Nishida J, Wake N,, K-Ras and H-Ras activation promote distinct consequences on endometrial cell survival., Cancer Research, 10.1158/0008-5472.CAN-3487-2, 64, 8, 2759-2765, 64, 2759-2765, 2004.04.
146. Kamikihara T, Arima T, Kato K, Matsuda T, Kato H, Douchi T, Nagata Y, Wake N, Epigenetic silencing of the imprinted gene ZAC by DNA methylation is an  early event in the progression of human ovarian cancer., International Journal of Cancer., 10.1002/ijc.20971, 115, 5, 690-700, 115,690-700, 2005.03.
147. Horiuchi S, Kato K, Suga S, Takahashi A, Ueoka Y, Arima T, Nishida JI, Hachisuga T, Kawarabayashi T, Wake N., Expression of progesterone receptor B is associated with G0/G1 arrest of the cell cycle and growth inhibition in NIH3T3 cells., Exp Cell Res, 10.1016/j.yexcr.2005.01.003, 305, 2, 233-243, 1;305(2):233-243., 2005.05.