| 1. |
Y Miyazaki, T Satoyama, H Nakano, S Takeuchi, H Takeuchi, D Tachikawa, The anti-inflammatic activity of a sulfated polysaccharide Fucoidan in innate immune cells, 106th Annual Meeting of The American Association of Immunologists, 2022.05, Fucoidan is a series of natural sulfated polysaccharides derived from brown seaweeds and have reported to possess various biological activities such as anti-tumor, anti-virus, and immuno-modulatory effects. In previous study, we revealed that fucoidan derived from Cladosiphon okamuranus activated murine macrophage-like cell line RAW264 cooperatively with Zymosan, a Saccharomyces cerevisiae-derived β-glucan. On the other hand, several reports demonstrated that fucoidan inhibited lipopolysaccharide (LPS)-induced production of nitrogen oxide (NO) and inflammatory cytokines by RAW264 cells. Therefore, in this study, we investigated the potential of fucoidan derived from Undaria pinnatifida to regulate the excessive activation of RAW264 cells in response to overstimulation with pathogen components. RAW264 cells were inoculated at 20000 cells/well in 96-well culture plates, and after a recovery culture for 24 hr, the cells were treated with fucoidan with or without appropriate concentrations of each ligand for pattern recognition receptors (PRRs) including Toll-like receptors (TLRs). Whereas the production of NO was significantly enhanced with fucoidan alone, it was dose-dependently inhibited by fucoidan under stimulation with Pam3CSK4 (TLR1/TLR2 ligand), heat killed Listeria monocytogenes (HKLM, TLR2 ligand), LPS (TLR4 ligand), Pam2CGDPKHPKSF (FSL-1, TLR2/TLR6 ligand). These results suggested that fucoidan have beneficial ability to alleviate excessive inflammatory reaction during pathogenic infection and maintain suitable immune balance.. |
| 2. |
大野波月, 中野勇人, 柴崎哲哉, 里山俊哉, 宮﨑義之, 中山二郎, in vitro実験系を用いたフコイダンのヒト腸内細菌叢に対する効果の解析, 日本農芸化学会, 2022.03. |
| 3. |
宮﨑義之，朴俊夏，中野勇人，竹内秀吾，竹内秀晃, 立川大介, Anti-angiogenic effect of fucoidan-mix AG via improvement of tumor microenvironment in a mouse melanoma model, 第50回日本免疫学会, 2021.12, Fucoidan is a series of natural sulfated polysaccharides constitute viscous component of brown seaweeds. Previously, we reported that a mixed preparation consists of high-molecular weight fucoidans derived from Okinawa-mozuku and Mekabu (sporophyll of Wakame) and Agaricus blazei mycelium extract, which named fucoidan-mix AG, acted to reinforce anti-tumor immune functions. In this study, we evaluated dietary effects of fucoidan-mix AG on tumor angiogenesis with C57BL/6 mice subcutaneously implanted basement membrane matrix (GeltrexTM) enclosing 5x104 mitomycin C-treated B16 melanoma cells or 500 ng/mL vascular endothelial growth factor (VEGF), and 2% fucoidan-mix AG containing diet was fed for 7 days before and after the matrix injection. As the result, accumulation of hemoglobin (Hb) and CD31-positive cells in the B16- enclosed matrix was lower by the fucoidan-mix AG feeding, which indicated that fucoidan-mix AG was effective to suppress tumor angiogenesis. In contrast, the Hb contents in the VEGF- enclosed matrix were not affected by the fucoidan-mix AG intake. Because the VEGF gene expression in the B16-enclosed matrix was reduced in the fucoidan-mix AG-fed mice, it was guessed that fucoidan-mix AG did not inhibit angiogenic activity of VEGF but suppressed VEGF production. In this context, the matrix infiltration of CD206-positive macrophages, which known to produce VEGF, were declined in the fucoidan-mix AG-fed mice. Furthermore, fucoidan-mix AG decreased matrix accumulation of Treg, and delayed B16 tumor growth. In conclusion, fucoidan-mix AG was expected to be effective in inhibition of tumor angiogenesis by improving immunosuppressive tumor microenvironment.. |
| 4. |
宮﨑義之，朴俊夏，中野勇人，竹内秀吾，竹内秀晃, 立川大介, がん性血管新生および腫瘍の形成に及ぼすフコイダン− アガリクスミックス(CUA-フコイダン)の摂食効果, 第68回日本食品科学工学会, 2021.08, 【目的】 フコイダンは，L-フコースを主な構成糖とする褐藻類由来の天然硫酸 化多糖であり，抗がん作用や抗ウイルス作用などの生理活性が報告されている. 先の研究で我々は, オキナワモズクおよびメカブに由来する2種類のフコイダン とアガリクス菌糸体エキス末を配合したフコイダン-アガリクスミックス(通称: CUA-フコイダン)が，免疫増強作用を通してがん腫瘍形成を効果的に抑制する ことを明らかにしてきた.本研究では，新たな視点としてがん性血管新生に着目 し CUA-フコイダンの摂食効果とその作用機序を検証した.
【方法】 本試験では C57BL/6 マウス(n=6)を用い，マイトマイシン C 処理により 増殖を阻害した 5x104 個の B16 メラノーマ細胞または 500 ng/mL の血管内皮細 胞増殖因子(VEGF)を封入した基底膜マトリクス(GeltrexTM)をマウスの背部皮 下に注射した.マトリクス皮下注射の前後 7 日間にわたり CUA-フコイダンを 2% 重量添加した AIN-93G 準拠食を自由摂食させ，マトリクスゲル内での血管形成 (ヘモグロビン(Hb)蓄積，血管内皮細胞数)および免疫細胞浸潤を評価した. 【結果】 B16 細胞を封入したマトリクスゲルにおける Hb 蓄積量は，CUA-フコイ ダン摂取群で有意に低値を示し，CD31 陽性血管内皮細胞の割合もまた CUA- フコイダン摂取群で減少傾向が観察された.これらの結果から，CUA-フコイダ ンはがん性血管新生に対して抑制作用を有することが示された.一方，VEGF を封入したマトリクスゲルでの Hb 蓄積には CUA-フコイダン摂取の効果は観察 されず，VEGF の血管新生誘導作用には影響を及ぼさないことが示された.し かし，ゲル中の VEGF 遺伝子発現は CUA-フコイダン摂取群で低値を示してお り，血管新生抑制の作用機序の一つとして，腫瘍内で VEGF を産生する CD206 陽性マクロファージの浸潤が CUA-フコイダン摂取に伴い減少することが考えら れた.さらに，CUA-フコイダンが抗腫瘍免疫の抑制に働く制御性 T 細胞(Treg) の浸潤減少や B16 腫瘍の成長遅延に働く可能性が併せて示された.以上の結 果から，CUA-フコイダンは腫瘍内の免疫抑制的な環境を改善し，がん性血管 新生および腫瘍形成の効果的な阻害に寄与することが期待された.. |
| 5. |
Y Miyazaki, J Bak, H Nakano, S Takeuchi, H Takeuchi, D Tachikawa, Regulatory mechanism of fucoidan-mix AG in intestinal mucosal immune cells, 105th Annual Meeting of The American Association of Immunologists, 2021.05, Fucoidan is a series of natural sulfated polysaccharides contained in brown sea algae, and their beneficial physiological activities such as anti-tumor, anti-virus and immune regulatory effects have been reported. In previous studies, we proved that fucoidans derived from Cladosiphon okamuranus (C. okamuranus) and Undaria pinnatifida effectively improved anti-tumor immunity and tumor vaccine efficiency in combination with an Agaricus blazei mycelium extract. It is thought that the fucoidan-agaricus mix (named fucoidan-mix AG) stimulate innate immune cells to activate intestinal and mucosal immune reaction, but actual target immune tissues and cells by fucoidan have not been evidenced yet. In this study, we investigated action mechanisms of fucoidan in regulation of mucosal immune activities. At first, immune cells were isolated from Payer’s patches, mesenteric lymph node (MLN) and intestinal epithelia of Balb/c mice, then stained with fluorophore-labeled fucoidan. As the results, it was revealed that fucoidan directly interact with lymphocytes besides dendritic cells and macrophages existing in these intestinal immune tissues. Furthermore, immune cells from Payer’s patches and MLN stimulated with fucoidan from C. okamuranus and fucoidan-mix AG produced more TNF-α, IL-12 and Th1 cytokine IFN-γ. The production of an immunosuppressive cytokine IL-10 also was augmented in the fucoidan-treated immune cells, which suggesting that fucoidan balanced immune system to prevent occurrence of excessive reaction. In addition, it was newly proved that fucoidan directly stimulated T lymphocytes to induce enhanced IFN-γ production.. |
| 6. |
篠田あかり, 朴俊夏, 中野勇人, 柴崎哲哉, 宮﨑義之, 中山二郎, 腸内環境および免疫機能に及ぼすあまおう乳酸菌およびフコイダンの摂食効果, 日本農芸化学会, 2021.03. |
| 7. |
Y Miyazaki, Immune-improving activities of fucoidan, seaweed-derived natural sulfated polysaccharides, 2021 Health Food Society of Taiwan Annual Meeting & International Symposium on Health Food Development in the Post-Pandemic Era, 2021.03, Fucoidan is one of intercellular viscous components cover surface of brown sea seaweeds and a series of natural sulfated polysaccharides. Peoples in Asian countries eat seaweeds willingly as nourishing healthy foods, and fucoidan have been reported to have beneficial physiological activities such as anti-tumor, anti-virus and immune regulatory effects. In this context, we have proved in mouse model experiments that fucoidans derived from Cladosiphon okamuranus (Okinawa-mozuku) and Undaria pinnatifida (Mekabu) in combination with an Agaricus blazei mycelium extract effectively improved anti-tumor immunity, and potentially support effective induction of anti-tumor immune function by vaccination with tumor antigen peptide. Furthermore, we conducted open-label trials with the fucoidan-agaricus mix (named fucoidan-mix AG), and revealed that its intake was effective to augment natural killer (NK) cell activities and salivary secretory immunoglobulin A (sIgA) production in humans. On the other hand, we evaluated the capacity of Cladoshiphon okamuranus-derived fucoidan to activate murine macrophage-like cell line RAW264, and investigated the functional relationship with a Saccharomyces cerevisiae-derived β-glucan. As the results, it was shown that the interaction of fucoidan on cell surface activated RAW264 cells to produce more nitric oxide and TNF-α, and the maximum augmentation was observed in combinational stimulation with fucoidan and β-glucan. The results suggested that fucoidan and β-glucan had a cooperative effect to reinforce macrophage function. In conclusion, these results indicate that the intake of fucoidan is useful in augmentation of intestinal and mucosal immune defense, and expects to contribute to the prevention of cancer and infectious diseases and support our healthy life.. |
| 8. |
Y Miyazaki, J Bak, H Nakano, S Takeuchi, H Takeuchi, D Tachikawa, Suppressive effects of fucoidan-agaricus mix (CUA-fucoidan) on angiogenesis and tumor growth in B16 melanoma-bearing mice, 104th Annual Meeting of The American Association of Immunologists, 2020.05. |
| 9. |
Juneha Bak, Yoshiyuki Miyazaki, Toshiro Matsui, An establishment of NMR-based analytical assay for sulfated saccharides in solution, ICoFF2019/ISNFF2019 KOBE, 2019.12. |
| 10. |
Juneha Bak, Yoshiyuki Miyazaki, Toshiro Matsui, Ligand-aided quantitative and selective NMR assay for sulfated saccharides in solution., International symposium between National Taiwan University and Kyushu University, 2019.12. |
| 11. |
祝原由莉，中溝公次，竹内秀吾，竹内秀晃, 立川大介, 宮﨑義之, 硫酸化多糖フコイダンによるマクロファージ活性化機構の解明, 第65回日本食品科学工学会大会, 2018.08. |
| 12. |
Y Miyazaki, Y Iwaihara, M Nakamizo, S Takeuchi, H Takeuchi, D Tachikawa, Potentiating effects of high-molecular weight fucoidan-agaricus mix (CUA) feeding on tumor vaccination, 102nd Annual Meeting of The American Association of Immunologists, 2018.05. |
| 13. |
宮﨑義之, 祝原由莉, 中溝公次, 竹内秀伍, 竹内秀晃, 立川大介, The induction of macrophage activation by a sulfated polysaccharide fucoidan, 第46回日本免疫学会, 2017.12. |
| 14. |
宮﨑 義之, 中溝 公次, 川原 和弘, 竹内 英明, 立川 大介, 山田 耕路, Augmentation of salivary IgA production in healthy subjects by fucoidan-agaricus mix ingestion, The American Association of Immunologists, 2016.05, Fucoidan is a series of sulfated polysaccharides derived from brown algae, and have reported to have various biological activities. Previously, we demonstrated that fucoidan derived from Cladosiphon okamuranus and Undaria pinnatifida effectively augmented anti-tumor immunity in combination with Agaricus blazei mycelia extract. In this study, we evaluated the capacity of the fucoidan-agaricus mix (FAM) to enhance oral mucosal immune function. Fifteen healthy volunteers (mean age, 41.2 years old; range, 22-56 years old; 8 males, 7 females) ingested 4 capsules each containing 250 mg FAM powder every day for 12 weeks. As a result, the mean of secretory rates of salivary secretory immunoglobulin A (sIgA) tended to be increased by the FAM administration for 4 and 12 weeks as compared with the initial value. The enhanced salivary sIgA secretion was more distinctly observed in subject group whose initial values of salivary sIgA secretory rate were lower than the total average. Furthermore, the intake of FAM led to significant augmentation in the salivary sIgA secretion (by 1.3-fold) in the group of subjects under 40 years of age. On the other hand, concanavalin A-induced blastogenic response of peripheral blood mononuclear cells and serum IgA concentration was not elevated during this trial. Therefore, it was suggested that FAM stimulated functional maturation rather than expansion of B lymphocyte. The intake of FAM did not significantly affect NK cell activities. In addition, the safety of FAM consumption was confirmed because no abnormal findings were observed in general clinical tests. From the results, it was suggested that intake of FAM was useful in augmentation of oral mucosal immune defense via enhancing salivary sIgA production.. |
| 15. |
宮﨑 義之, 中溝 公次, 桐野 智美, 山口 千仁, 立花 宏文, 山田 耕路, Pilot clinical study to evaluate the immune enhancing activity of fucoidan mix, 第12回アジア栄養学会議, 2015.05, Objectives: Fucoidan, sulfated polysaccharides from brown algae, have reported to have immunomodulatory effects, however the validation in human use is not enough. In this study, we performed an open label trial to evaluate the capability of a fucoidan-containing healthy food (fucoidan mix) to enhance immune function. Materials & Methods: The fucoidan mix is a powdered food blending Okinawamozuku fucoidan, Mekabu fucoidan and Agaricus mycelium extract. Fifteen healthy volunteers (mean age, 41.2 years; range, 22-56 years; 8 male, 7 female) were ingested 4 capsules containing 250 mg of fucoidan mix every day for 12 weeks. Immunological parameters including salivary secretory immunoglobulin A (sIgA) secretory rates, concanavalin A-induced blastogenic responses and NK cell activities of peripheral blood mononuclear cells were measured at before and 4, 8, 12 weeks after the fucoidan mix ingestion.
Results & Findings: First of all, the mean of secretory rates of salivary sIgA tended to be increased by the fucoidan mix administration for 4 and 12 weeks as compared with the initial value. The enhanced secretion of sIgA was more distinctly observed in subject group whose initial values on sIgA secretory rate were lower than the whole average. Furthermore, fucoidan mix significantly augmented the sIgA secretion (by 1.3-fold) in the group of subjects under 40 years of age. On the other hand, the lymphocyte blastogenic response was not reinforced during this trial, and it was suggested that fucoidan mix stimulated rather functional maturation than expansion of B lymphocyte. The intake of fucoidan mix did not significantly affect NK cell activities. In addition, the safety of fucoidan mix was confirmed, because no abnormal findings were observed in general clinical tests.
Conclusion: From the results, it was suggested that oral administration of fucoidan mix was useful in augmentation of oral mucosal immune defense through enhancement of salivary sIgA production.. |
| 16. |
宮﨑 義之, 中溝 公次, 桐野 智美, 立川 大介, 立花 宏文, 山田 耕路, フコイダン-アガリクスミックス経口摂取による抗がん剤投与に伴う免疫抑制の緩和 〜マウスモデル試験による検証〜, 日本食品科学工学会, 2014.08, 【目的】 フコイダンは、主に L-フコースから成る褐藻由来の硫酸化多糖類であ る。フコイダンは、抗がん作用や免疫増強作用などの様々な生理活性を有する ことが多く報告されているが、抗がん剤治療と併用することの有用性は十分に検 討されていない。本研究では、オキナワモズクおよびメカブ由来のフコイダン抽 出物にアガリクスエキス末を混合した調製品(Fucoidan-Agaricus Mix, FAM)を 用い、抗がん剤の副作用である免疫抑制に対する緩和効果を検討した。 【方法】 実験には、Balb/c 雄マウスを用い、抗がん剤 5-フルオロウラシル (5-FU)をマウス 1 頭あたり 1 mg、隔日5回皮下注射することで、抗がん剤治療 モデル試験系を構築した。比較対照として、同量のリン酸緩衝生理食塩水 (PBS)を皮下注射した。また、5-FU 投与および非投与マウスには、それぞれ、1 日あたり 0, 20 または 40 mg の FAM を含む餌を、5-FU 投与開始の 3 日前から 最終投与の 2 日後まで与えた。その後、採血ならびに脾臓を採取し、脾臓細胞 の増殖能、サイトカイン産生能および免疫細胞集団の割合などを評価した。 【結果】 始めに、抗がん剤投与の効果を検討するために血中ヘモグロビン量お よび脾臓重量を測定した結果、共に 5-FU を投与したマウスで有意な低下が観 察され、抗がん剤副作用による骨髄抑制および免疫抑制の評価に適したモデ ル実験系であることが示唆された。また、本モデルマウスでは、がん細胞の殺傷 に働く NK 細胞の活性が 5-FU 投与によって有意に低下したが、FAM 摂取によ って用量依存的に上昇し、40 mg FAM 摂取群で正常値まで回復した。また、 NK 細胞などの抗腫瘍免疫に関わる細胞の活性化を誘導するインターフェロン -γ(IFN-γ)についても、5-FU 投与による低下が、FAM 摂取によって顕著に軽減 された。さらに、免疫細胞の存在比をフローサイトメーターで測定した結果、 IFN-γ を産生する Th1 細胞や抗腫瘍免疫の亢進に働くマクロファージなどで、 FAM 摂取による 5-FU 細胞毒性の緩和が観察された。以上の結果から、FAM 摂取が、抗がん剤 5-FU の副作用の一つである抗腫瘍免疫機能の低下の緩和 に有効であることが示唆された。. |
| 17. |
宮﨑 義之, 中溝 公次, 柴崎 哲哉, 桐野 智美, 川原 和弘, 竹内 英明, 立花 宏文, 山田 耕路, 立川 大介, Immune enhancing effects of fucoidan-agaricus mix under treatment of an immunosuppressive anti-cancer agent, The American Association of Immunologists, 2014.05, Fucoidan is a series of sulfated polysaccharides derived from brown algae that mainly consist of L-fucose. Fucoidan have various biological activities, and our investigation with experimental animal models and a healthy human trial demonstrated fucoidan from Cladosiphon okamuranus and Undaria pinnatifida effectively augmented anti-tumor immunity in combination with agaricus mycelium extract. In this context, we have reported that feeding of the fucoidan-agaricus mix (FAM) protected mice from immunosuppressive effect of 5-fluorouracil (5-FU). In current study, we confirmed dose-dependent immunoprotective effects of the dietary FAM (at 20 and 40 mg/day) by observing recovery in spleen weight, NK cell activity and IFN-γ production in 5-FU-treated mice. Furthermore, we investigated in vitro effects of FAM against 5-FU on growth and function of primary cultured immune-related cells. As the results with thioglycollate-induced peritoneal exudates cells (PEC), 5-FU dose-dependently suppressed the growth whereas FAM exerted no effects. However, production of cytokines such as IL-12p40 and TNF-α in PEC were stimulated by FAM treatment even in the presence of 5-FU. In the case of bone marrow derived cells differentiated with GM-CSF and M-CSF, FAM markedly enhanced the growth and IL-12 production of these cells. From these results, it was suggested that FAM contributed to amelioration of decline in anti-tumor immune function by 5-FU treatment through activation of innate immune cells.. |
| 18. |
宮﨑 義之, 中溝 公次, 柴崎 哲哉, 桐野 智美, 齋藤 ゆず, 川原 和弘, 立花 宏文, 山田 耕路, 立川 大介, Protective effects of orally administered fucoidan mix on immunosuppression by an experimental cancer chemotherapy, The American Association of Immunologists, 2013.05, Fucoidan are sulfated polysaccharides derived from brown seaweeds that mainly consist of L-fucose. Peoples in Asian countries eat seaweeds willingly as healthy foods. In previous studies, we have demonstrated that fucoidan augment anti-tumor immunity in an experimental mouse model and in healthy human subjects. However, availability of fucoidan to protect immune system from hazardous side effects of cancer chemotherapy has not been verified enough. Therefore, we performed a mouse model experiment in which immunosuppression was induced by subcutaneous 5-fluorouracil (5-FU) injection (1 mg/mouse/day, 5 times on alternate days) and evaluated protective effects of orally administered fucoidan mix (40 mg/mouse/day). The immunosuppression by 5-FU s.c. was confirmed with the loss in spleen weight of 5-FU-injected mice. NK cell activity was lower in 5-FU-injected mice than sham-treated control mice, but the decline was recovered by the fucoidan mix feeding. The intake of fucoidan mix significantly alleviated the reduction in interferon (IFN)-gamma production of splenocytes from 5-FU-injected mice. Furthermore, flow cytometric analyses revealed that specific population such as Th1 and CD11b+ cells were supported by the fucoidan mix feeding even in the 5-FU-injected mice. These results suggested that oral administration of fucoidan mix is effective to maintain tumor immunity during immunosuppressive cancer chemotherapy.. |
| 19. |
宮﨑 義之, 中溝 公次, 辻 博光, 桐野 智美, 齋藤 ゆず, 川原 和弘, 立花 宏文, 山田 耕路, 藤井 信, 立川 大介, Enhancement of NK cell activity and Th1 immunity in healthy subjects by orally administered fucoidan mix, The American Association of Immunologists, 2012.05, Fucoidan is a sulfated polysaccharide derived from brown seaweeds which mainly consist of L-fucose. In Asian countries such as Japan, Korea and China, peoples eat seaweeds willingly as nourishing health foods. We previously showed that the tumor formation by transplanted Sarcoma180 cells was suppressed in mice fed 2% fucoidan mix containing diets, and NK cell activity was enhanced in the fucoidan-fed mice. The results suggest that the activated NK cells participated in elimination of the tumor. However, in human, immune enhancing activities of fucoidan have hardly been proved. Therefore, we performed an open-label trial in this study. Five healthy volunteers (47 ± 6 years old, 2 male and 3 female) took 2.5 g fucoidan mix per day for 30 days. There were no abnormal observations in the values of WBC, RBC, platelet count and blood profile. On the other hand, NK cell activity was augmented by 1.42 times in the average at end of the trial than the initial value. The ratio of IFN-gamma-producing (Th1) cells also increased to 114% for the initial value by the day 30 by the fucoidan mix intake, whereas IL-4-producing (Th2) cells decreased to 77%. As a result, the rise of Th1/Th2 ratio by 1.46 times in the average was observed with 4 subjects among 5 examinees. Furthermore, CD8+ cells tended to expand with the all subjects. These results suggest that intake of fucoidan is effective to augment anti-tumor immunity in human.. |
| 20. |
宮﨑 義之, 山﨑（宮本）有美, 立花 宏文, 山田 耕路, Biological functions and anti-cancer effect of fucoidan from brown algae, International Society for Nutraceuticals & Functional Foods, 2010.10, Fucoidan is one of viscous components from brown seaweeds including Cladosiphon okamuranus TOKIDA (Okinawamozuku), Kjellmaniella crassifolia (Gagome) and Undaria pinnatifida (Mekabu), and is collective term for sulfated polysaccharides which mainly consist of L-fucose. At present, fucoidan is known to have several biological activities such as anti-pathogenic, anti-oxidant, and anti-inflammatory activities. In particular, we focus on the anti-tumor activity of fucoidan, and have revealed that Okinawamozuku fucoidan induced apoptotic cell death of a human breast cancer cell line, MCF-7, by activating intracellular signaling cascade through caspase-8, (-9) and -7. Other plausible mechanisms for anti-tumor action of fucoidan are inhibition of angiogenesis, and buildup of anti-tumor immunity. It is thought that fucoidan stimulate innate immune cells existing in Peyer’s patch to induce production of cytokines including IFN-γ and IL-12, and they activate effector cells such as natural killer cells. Further experiments will surely provide useful information for applying fucoidan to cancer therapies.. |
