BACKGROUND:Autophagy,as a key regulatory mechanism of cell development,plays an important role in different stages of embryonic development.The mechanism of how autophagy regulates embryonic development through histone modifications is currently unclear.OBJECTIVE:To investigate the effect of autophagy on trimethylation of lysine 4 on histone H3(H3K4me3)modification in embryos and its effect on embryonic development.METHODS:Mouse fertilized eggs were divided into control and autophagy inhibitor-treated groups(chloroquine phosphate-treated group and 3-methyladenine-treated group),and cultured in vitro to different periods of time,and were then classified as early 2-cell embryos,middle 2-cell embryos,late 2-cell embryos,4-cell embryos,8-cell embryos,morula stage,and blastocyst stage.Levels of reactive oxygen species,autophagy marker proteins LC3B and P62,DNA loss marker γH2AX,and H3K4me3 were analyzed by immunofluorescence assay in late 2-cell embryos of each group.Changes in H3K4me3 modification in late 2-cell embryos of each group were detected by CUT&Tag.RESULTS AND CONCLUSION:(1)Autophagy inhibition caused embryo development arrest.(2)There was no significant difference in reactive oxygen species and γH2AX between the autophagy inhibitor-treated groups and control group.(3)H3K4me3 levels were significantly elevated in the autophagy inhibitor-treated group compared with the control group.(4)CUT&Tag results showed a significantly increased H3K4me3 peaks on the proximal promoter region of the genes after autophagy inhibition and an increase of H3K4me3-specific modification genes.These findings suggest that autophagy may affect embryonic development by regulating the level of H3K4me3 modification.

BACKGROUND:Exercise is an effective way to improve microvascular function in patients with type 2 diabetes.In recent years,exercise has been used as an intervention therapy for microvascular dysfunction in patients with type 2 diabetes.However,few studies have systematically explored the influence of factors,such as"exercise type,exercise intensity and amount of exercise,"on microvascular function in patients.To some extent,this limits the formulation of precise exercise prescriptions to improve microvascular dysfunction in patients with type 2 diabetes and the comparison of study results.OBJECTIVE:To investigate the effects of exercise type,intensity,frequency and amount of exercise on microvascular function in patients with type 2 diabetes mellitus,and to make suggestions on exercise prescription.METHODS:The first author used computer to search the studies on the improvement of microvascular function in type 2 diabetes patients involving exercise in CNKI,WanFang,PubMed and other databases.The search terms were"diabetes mellitus,type 2 diabetes mellitus,microcirculation,microvascular reactivity,microvessels,capillaries,vasodilation,blood perfusion volume,endothelial cells,shear stress,exercise,aerobic exercise,resistance exercise,high-intensity exercise"in Chinese and English.The articles were screened by a quick glance at the article titles and abstracts to exclude those that were not closely related to the topic,and finally 60 articles were included for review.RESULTS AND CONCLUSION:(1)Exercise is an effective way to improve microvascular function in patients with type 2 diabetes.Aerobic exercise lasting 12-24 weeks,3-5 times/week,exercise time＞30 minutes and intensity between 40％and 59％reserve oxygen intake can significantly improve microvascular function in patients with type 2 diabetes.On the basis of aerobic exercise,systemic resistance exercise 2-3 times a week(50％-85％1RM,every other day)or pressure resistance exercise can obtain better intervention effects.(2)In addition,exercise can improve microvascular function in patients with type 2 diabetes in a"dose-effect"manner,and patients can get better results from the intervention by increasing the amount of exercise,while maintaining safety.(3)The mechanism of exercise improving microvascular function in patients with type 2 diabetes is mainly related to promoting the release of nitric oxide and vascular endothelial growth factor from endothelial cells and inhibiting the release of endothelin1.

Background: Chinese herbal pieces are an essential component of traditional Chinese medicine. Accurate identification and classification of these materials are crucial in clinical practice. Objective: This study aims to enhance the recognition efficiency of Chinese herbal pieces using deep learning technology, while addressing the limitations of traditional manual classification methods in terms of both quality and efficiency. Methods: A comprehensive dataset containing 201 types of Chinese herbal pieces was established. Based on Real-time Detection Transformer (RT-DETR), we designed and integrated a Feature-focused Diffusion Network (FDN), resulting in an improved model termed RT-DETR-FDN. The proposed FDN includes a Feature-focus Module and a feature diffusion mechanism, enabling the model to capture more extensive feature information from Chinese herbal pieces and diffuse it across multiple detection scales. Results: Experimental results show that RT-DETR-FDN achieved a precision of 0.925, a recall of 0.943, and an mAP50-95 of 0.851. In addition, the model was compared with representative You Only Look Once series models commonly used in object detection. Compared with these models, RT-DETR-FDN achieved higher recognition accuracy while maintaining a lightweight architecture. Conclusion: This study integrates deep learning with traditional Chinese medicine, providing a more effective solution for the recognition of Chinese herbal pieces.

Objective: To investigate the effect of the senescence state of parental human bone marrow mesenchymal stem cells (BMSCs) on the osteoinductive properties of their derived apoptotic vesicles (apoVs), and to provide an experimental basis and a quality control reference for the treatment of alveolar bone defects and osteoporosis based on apoVs. Methods: A replicative senescence model of human aging BMSCs (A-BMSCs) was established via serial passaging, with young BMSCs (Y-BMSCs) serving as controls. Following the induction of apoptosis with staurosporine, apoVs were isolated from Y-BMSCs and A-BMSCs (termed Y-apoVs and A-apoVs, respectively) via differential centrifugation. The physicochemical properties (morphology, size, zeta potential, and yield) and protein markers of both apoV populations were systematically characterized. Subsequently, Y-BMSCs were divided into a proliferation medium (PM) group, osteogenic induction medium (OM) group, OM + Y-apoVs group, and OM + A-apoVs group. The osteoinductive efficacy in vitro was evaluated by alkaline phosphatase (ALP) staining, alizarin red S staining, and quantitative real-time PCR detection of key osteogenic genes, including runt-related transcription factor 2 (RUNX2), ALP, osteopontin (OPN), and osterix (OSX). Approved by the Institutional Animal Care and Use Committee, in vivo biodistribution (labelling apoVs with the red fluorescent dye PKH26) and bone regeneration efficacy were assessed in 18-month-old osteoporotic C57BL/6 mice. Mice were randomly divided into a control group (injected with phosphate-buffered saline), Y-apoVs group (injected with Y-apoVs), and A-apoVs group (injected with A-apoVs). Following tail-vein injection administration of apoVs for 8 weeks, bone regeneration was evaluated via micro-computed tomography and histological analysis. Results: Electron microscopy and particle size analysis revealed that both Y-apoVs and A-apoVs displayed typical biconcave discoid structures, with diameters mainly ranging from 100 to 500 nm. Western blot assays confirmed high expression of universal vesicle markers (CD9, CD63, CD81) and the apoptotic marker Fas in both groups. Y-apoVs and A-apoVs exhibited indistinguishable morphologies, size distributions, zeta potentials, and yields. In vitro experiments showed that, compared with the PM group, the OM, OM + A-apoVs, and OM + Y-apoVs groups significantly enhanced ALP activity, calcium nodule formation, and the expression of osteogenic genes (RUNX2, ALP, OPN, OSX) in recipient Y-BMSCs (P<0.05); moreover, compared with the OM and OM + A-apoVs groups, the OM + Y-apoVs group exhibited a more significant promoting effect (P<0.05). An in vivo analysis demonstrated that tail-vein-injected apoVs effectively homed to bone tissue. Moreover, the Y-apoVs group significantly improved trabecular microarchitecture, bone mineral density, and bone volume fraction in aged mice, exhibiting superior therapeutic efficacy over the control and A-apoVs groups (P<0.05) Conclusion The senescence state of parental cells significantly impairs the osteoinductive ability of BMSC-derived apoVs. Y-apoVs are effective biological agents for the treatment of age-related bone loss.

[摘 要] 目的：探讨蟾毒灵（BU）对肿瘤相关巨噬细胞（TAM）介导的结直肠癌（CRC）细胞程序性死亡蛋白-配体1（PD-L1）表达的调控作用及其分子机制。方法：采用HCT116细胞与THP-1来源巨噬细胞构建体外共培养体系。佛波酯（PMA）诱导THP-1细胞分化为M0型巨噬细胞，进一步以HCT116细胞条件培养基（CM）刺激形成TAM样细胞。流式细胞术检测CD11b、CD206表达，以评价巨噬细胞极化水平；RT-qPCR及WB法检测TGF-β、IL-10、STAT3/p-STAT3及PD-L1表达变化；慢病毒转染构建STAT3敲低细胞系HCT116sh-STAT3，结合BU干预验证STAT3/PD-L1信号通路作用。结果：HCT116细胞CM可诱导巨噬细胞向M2表型极化，表现为CD11b⁺CD206⁺细胞比例升高（P < 0.01）及TGF-β、IL-10表达增加（均P < 0.001）。TAM条件培养基可显著促进HCT116细胞STAT3磷酸化（P < 0.001）并上调PD-L1 mRNA及蛋白表达水平（P < 0.001或P < 0.01）。BU干预后，TAM介导的STAT3磷酸化水平明显下降，PD-L1表达同步下调（均P < 0.01）。STAT3敲低可降低PD-L1表达，其作用趋势与BU一致。结论：BU可通过抑制TAM介导的STAT3信号激活，下调CRC细胞PD-L1表达，提示其在肿瘤免疫微环境调控中具有潜在的应用价值。

[摘 要] 目的：探究西贝素通过调控Cyclin D1介导的CDK4/6-Rb-E2F1信号轴对人胃腺癌细胞（AGS）的增殖抑制效果及其分子机制。方法：利用不同浓度的西贝素处理AGS和人正常胃黏膜上皮细胞（GES-1），通过CCK-8检测和结晶紫染色实验确定西贝素的最适抑制剂量和作用时间；采用划痕实验和Transwell迁移实验与侵袭实验检测西贝素对AGS细胞迁移与侵袭能力的影响；利用流式细胞术和WB检测西贝素对AGS细胞周期及细胞周期相关蛋白表达的影响；利用质粒过表达Cyclin D1，并通过WB检测、结晶紫染色实验、划痕实验和Transwell迁移实验与侵袭实验进一步明确西贝素的抑制机制；构建AGS移植瘤裸鼠模型，采用免疫荧光染色法和免疫组化检测Cyclin D1、CDK4、CDK6、Rb、p-Rb、E2F1的表达水平。结果：与对照组相比，西贝素药物浓度为100 μg/mL时，对GES-1细胞的增殖无影响（P > 0.05），但能够显著性抑制AGS细胞增殖、迁移和侵袭能力（P < 0.01），并诱导AGS细胞周期G0/G1期阻滞，显著抑制周期相关蛋白Cyclin D1、CDK4、CDK6、p-Rb、E2F1的表达水平（P < 0.05或P < 0.01）；Cyclin D1过表达能够显著降低西贝素对AGS细胞的抑制效果（P < 0.05或P < 0.01）；荷瘤裸鼠实验结果显示，西贝素能够抑制AGS移植瘤的增殖，影响肿瘤组织中Cyclin D1、CDK4、CDK6、p-Rb、E2F1的表达（P < 0.01）。结论：西贝素可能通过下调Cyclin D1介导的CDK4/6-Rb-E2F1信号轴活化抑制AGS增殖。

The brain-gut interaction theory is a multidimensional integrative concept based on the brain-gut axis， involving neural， endocrine， and immune regulatory networks as well as the gut microbiota. Zang-fu organs （脏腑） theory in traditional Chinese medicine （TCM） shows a high degree of consistency with the brain-gut interaction theory， and the core functions such as the spleen and stomach governing the ascending of the clear and descending of the turbid， the liver governing the free flow of qi， and the heart governing mental and emotional activities are closely associated with the multi-level regulatory mechanisms of the brain-gut axis. TCM therapy can modulate brain-gut interactions through multiple pathways in the treatment of spleen and stomach diseases， including the regulation of gastrointestinal hormone secretion， neurotransmitter levels， the hypothalamic-pituitary-adrenal （HPA） axis， immune homeostasis and inflammatory responses， as well as the gut microecology. However， current basic research on the brain-gut interaction theory in TCM for spleen and stomach diseases still faces several challenges， such as difficulties in integrating TCM spleen-stomach theory with modern pathophysiology， lack of innovation in research concepts， and limitations in research methodologies. It is therefore proposed that multidisciplinary collaboration， multi-omics technologies， and targeted research approaches should be adopted to provide more comprehensive methods for basic research on TCM spleen and stomach diseases， thereby promoting the in-depth development of brain-gut interaction theory.

This paper explores the differentiation and treatment strategies for pediatric IgA vasculitis based on the correlation between blood trubidity theory and oxidative stress. It is proposed that pediatric IgA vasculitis follows an evolution of pathogenesis characterized by "deficiency of healthy qi leading to turbidity generation， accumulation of turbid toxin， and toxin damage to the collateral vessels"， which corresponds to the pathological process of oxidative stress， namely decreased antioxidant capacity with accumulation of reactive oxygen species， metabolite deposition， and endothelial cell injury. A staged treatment strategy is proposed. In the acute stage， wind-toxin invading the colla-terals and reckless movement of heat in the blood are the main manifestations， for which the treatment should focus on dispelling wind and venting pathogens， resolving toxins and unblocking the collaterals， with a modified Yinqiao Powder （银翘散） and Xijiao Dihuang Decoction （犀角地黄汤） to regulate oxidative stress burst. In the prolonged stage， intermingling of turbidity and stasis with collateral obstruction is emphasized； treatment should focus on resolving turbidity and dispelling stasis， harmonizing the blood and stabilizing the collaterals， and a modified Simiao Powder （四妙散） and Taohong Siwu Decoction （桃红四物汤） can be used to improve microcirculatory dysfunction. In the remission stage， when healthy qi remains insufficient and residual pathogens persist， treatment should focus on strengthening the root and clearing the source， reinforcing healthy qi and nourishing the collaterals， for which modified Zhibai Dihuang Pill （知柏地黄丸） and Yupingfeng Powder （玉屏风散） is suggested to rebuild the antioxidant defense system.