Authorship：Lead author   Language：English   Publisher：Journal of Biological Chemistry  

5-Fluorouracil (5-FU) is a commonly used chemotherapeutic agent for breast cancer. Its efficacy relies on the function of p53, and mutations in p53 contribute to the development of resistance during 5-FU chemotherapy. Here, we report that microcurrent stimulation (MCS) of a p53-mutant breast cancer cell line induces p53-mediated cell death. Although MDA-MB-231 and MDA-MB-468 cells, both human breast cancer cell lines, are less sensitive to 5-FU due to p53 mutations, MCS (300 μA for 30 min) induced apoptosis in these cells and improved the antitumor effect of 5-FU in tumor-bearing mice. MCS-induced apoptosis was mediated by an increase in intracellular Cu²⁺ ions and reactive oxygen species, along with the concurrent transcriptional enhancement of pro-apoptotic genes by p53. Furthermore, MCS induced apoptosis in MDA-MB-231 cells that had developed resistance to 5-FU and inhibited tumor growth in tumor-bearing mice with reduced 5-FU sensitivity. These findings suggest that an approach involving MCS could serve as a foundation for developing breast cancer treatment strategies to overcome p53 mutations.

DOI： 10.1016/j.jbc.2025.110414

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Authorship：Lead author   Language：English   Publisher：Theranostics  

Rationale: Macrophage phagocytosis plays a role in cancer immunotherapy. The phagocytic activity of macrophages, regulated by circadian clock genes, shows time-dependent variation. Intervening in the circadian clock machinery of macrophages is a potentially novel approach to cancer immunotherapy; however, data on this approach are scarce. Microcurrent stimulation (MCS) promotes inflammation, proliferation, and remodeling, suggesting its potential to modulate macrophage function; however, its application has been limited. In this study, we investigated the impact of MCS on macrophage phagocytosis of cancer cells using mouse/human macrophage cell lines and various mouse/human cancer cell lines. Methods: Cells and mice received 300 µA, 400 Hz bidirectional pulsed MCS. Gene expression, protein expression, and phagocytosis activity were assessed in intraperitoneal macrophages collected from mice, as well as in RAW264.7, and THP-1 cells. Flow cytometry, population, phagocytosis activity, RNA-seq, and immunohistochemistry analyses were performed. Results: Noninvasive MCS prevented time-dependent reduction in macrophage phagocytosis of cancer cells by modulating the circadian clock genes. MCS also enhanced phagocytosis in mouse RAW264.7 and human THP-1 cells across various cancer types by promoting actin polymerization; similar in vivo effects were observed in mice. This enhancement occurred in abdominal macrophages of both sexes and was mediated by changes in clock gene expression. Specifically, suppressing the clock gene Per1 nullified the effects of MCS. Moreover, although macrophage phagocytosis typically declined during the dark period, MCS during the light period prevented this reduction. MCS also increased phagocytosis of peritoneally implanted cancer cells (4T1, ID8, and Hepa1-6) in mice, significantly reducing tumor engraftment and growth, and ultimately improving prognosis. Conclusions: The findings of this study suggest that targeting macrophage circadian mechanisms via MCS could enhance cancer immunity, offering new avenues for cancer immunotherapy.

DOI： 10.7150/thno.100748

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Language：English   Publisher：International Journal of Molecular Sciences  

The present review discusses vitamin A/retinoid metabolism as a cross-organ axis in which hepatic clock-dependent retinoid handling may affect immune clock gene expression through the stimulation of retinoic acid 6–Janus kinase 2–signal transducer and activator of transcription 5 signaling, potentially promoting pro-inflammatory monocyte states. We further highlight mechanosensory signaling as a second convergent layer that integrates hemodynamic forces with tissue microenvironmental cues. Among these pathways, G protein-coupled receptor 68, a proton- and flow-sensitive G protein-coupled receptor, is discussed as a representative druggable node linking mechanical and inflammatory signaling in chronic kidney disease-associated cardiac injury. Finally, we outline potential therapeutic directions, including (i) circadian alignment/chronopharmacology, (ii) modulation of retinoid metabolism and signaling, and (iii) targeted inhibition of primary immune and mechanosensory effectors.

DOI： 10.3390/ijms27083436

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Language：English   Publisher：Frontiers in Pharmacology  

Introduction – Hericenone C exhibits antinociceptive effects in inflammatory pain; however, its molecular target and underlying mechanism remain unclear. Methods and Results – We assessed the effect of hericenone C on formalin-induced nociceptive behavior in mice and explored its molecular target using in vitro experiments. Based on competitive affinity proteomics, we identified direct interactions between RORα and hericenone C; functional assays confirmed the role of hericenone C as a RORα antagonist that suppresses RORE-mediated transcriptional activity. Integrated bioinformatics and experimental validation indicated hericenone C-mediated suppression of TLR4 expression via inhibited RORα binding to the TLR4 promoter, which attenuates NF-κB signaling. This mechanism was further validated through pharmacological and genetic approaches, revealing that hericenone C and RORα antagonist SR3335 synergistically modulate TLR4 expression in RORα-modified macrophages. In the formalin-induced nociceptive pain model mice, formalin activated NF-κB through TLR4-dependent P65 phosphorylation, while macrophage depletion selectively suppressed phase 2 nociception. Critically, adoptive transfer of RORα-overexpressing or SR1078-pretreated monocyte-enriched PBMCs exacerbated pain, which was effectively reversed by hericenone C. Notably, hericenone C pretreatment reduced CD11c⁺ cell infiltration and decreased TLR4 expression in inflamed paw tissues. Conclusion – Overall, these findings establish hericenone C as a novel RORα antagonist that alleviates inflammatory pain through inhibition of the RORα-TLR4-NF-κB axis in CD11c⁺ cells, offering a promising therapeutic strategy for pain management.

DOI： 10.3389/fphar.2026.1703176

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Language：English   Publisher：Journal of Structural Biology  

The advent of direct electron detectors (DEDs) has driven a major breakthrough in cryo-electron microscopy (cryo-EM), particularly in single-particle analysis (SPA), establishing DEDs as essential tools for achieving near-atomic resolution. In this study, we re-evaluated the performance of the TVIPS TemCam-XF416, an indirect scintillator-coupled CMOS camera (scintillator camera). Using a JEOL CRYOARM 300II, we performed SPA on two well-established benchmark specimens, β-galactosidase and apoferritin, at a 300 kV acceleration voltage. The resulting reconstructions reached resolutions of 2.6 Å and 2.1 Å, respectively. Notably, the apoferritin map clearly resolves the central holes of aromatic side chains—a level of detail previously considered exclusive to DEDs. These results were achieved by implementing the latest standard reconstruction workflows, including motion correction and contrast transfer function refinement, underscoring the critical role of computational methods in attaining high-resolution structures. While scintillator cameras inherently exhibit a lower signal-to-noise ratio than DEDs, our findings with XF416 demonstrate that, with appropriate data collection and processing, such cameras can deliver near-atomic resolution structures. This work establishes a crucial technical benchmark for the scintillator camera evaluated in this study on a high-end 300 kV cryo-EM platform, demonstrating its capability to achieve resolutions suitable for many structural biology applications and providing an updated perspective on its performance capabilities.

DOI： 10.1016/j.jsb.2026.108286

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Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：Japanese