The nucleolus is an important nonmembranous organelle within the nucleus and is responsible for ribosome biogenesis. This process is frequently upregulated in tumors to support elevated protein synthesis and sustain tumor growth. The transcription of ribosomal DNA (rDNA) into pre-45S rRNA, a rate-limiting step in ribosomal biogenesis, has been reported to be enhanced to drive the progression of non-small cell lung cancer (NSCLC). However, the mechanism that upregulating rDNA transcription in NSCLC remains unclear. Here, we show that rDNA transcript levels were increased in NSCLC tumor tissues. Reduction of rDNA transcription by CX-5461, an RNA polymerase I inhibitor, significantly inhibited NSCLC cell proliferation. We found that the expression level of DDB1- and CUL4-associated factor 13 (DCAF13), a nucleolar protein, positively correlated with that of pre-45S rRNA. DCAF13 knockdown impairs rDNA transcription, ribosome biogenesis, and protein synthesis, indicating that DCAF13 participates in rDNA transcription. Mechanistically, DCAF13 interacts directly with TAF1A, a component of the RNA polymerase I preinitiation complex. This interaction is necessary for preinitiation complex assembly. In addition, DCAF13 is highly expressed in NSCLC, and its expression is negatively correlated with the overall survival in NSCLC patients. DCAF13 knockout inhibited NSCLC cell proliferation, colony formation, and migration, indicating that DCAF13 is a poor prognosis indicator of NSCLC. In summary, we found that DCAF13 promotes NSCLC cell proliferation by coordinating with TAF1A to regulate rDNA transcription and maintain high ribosome biogenesis.

Objective Karoshi, death from overwork, is a serious problem with unclear identification standards and mechanisms. This study aims to establish a karoshi rat model by integrating weight-bearing swimming and sleep deprivation. This model will enable us to investigate the adverse effects of acute physical and mental fatigue on cardiac functions and explore the response mechanisms to overwork using integrated omics approaches, specifically metabonomics and proteomics. Methods The experimental design involved healthy male sprague-dawley (SD) rats subjected to weight-bearing swimming and sleep deprivation for 7 d. The rats were monitored for changes in physiological function indexes, including electrocardiogram and respiration. Protein digestion, iTRAQ labeling, and quantitative data analyses were performed to determine differentially expressed proteins (DEPs). Additionally, GC-MS analysis was conducted to identify differential metabolites. The integration analysis of differential metabolites and proteins shared by the fatigue group and the overwork group was performed to construct a relevant metabolic pathway network and integrate the proteomics and metabolomics data. Statistical analysis was carried out using one-way ANOVA and Duncan's ' s multiple range t-tests. Results The rats subjected to weight-bearing swimming and sleep deprivation showed various physical and behavioral changes associated with fatigue, including hair disorder, decreased muscle tension, reduced food intake, and weight loss. Analysis of cardiac functions revealed cardiac hypertrophy and heart failure in the fatigue and karoshi groups, as evidenced by changes in heart color, myocardial fiber structure, heart rate, respiratory rate, and cardiac ultrasound measurements. Metabolomics analysis using GC-MS identified several differential metabolites in response to overwork, including amino acids involved in various metabolic pathways. Proteomic analysis using iTRAQ technology identified DEPs in the fatigue and karoshi groups, with a subset of DEPs shared by both groups. The GO analysis revealed that the up-regulated DEPs were primarily associated with mitochondria and peroxisomes in the cellular component category. The Reactome analysis further highlighted the enrichment of DEPs in the transfer of ferriheme from methemoglobin to hemopexin pathway. Integration analysis of the DEPs and differential metabolites revealed the activation of autophagy, increased mitochondrial oxidative phosphorylation, enhanced branched-chain amino acid degradation, and altered peroxisomal beta-oxidation. These findings suggested complex metabolic adaptations to meet the increased energy demands during overwork while also dealing with oxidative stress. Furthermore, the reprogramming of energy metabolism was observed, with upregulation of fatty acid beta-oxidation enzymes and glycolysis-related enzymes in the fatigue group, indicating a shift towards glucose metabolism. In contrast, the karoshi group showed a decreased dependence on fatty acids as an energy source and increased utilization of glucose. The model proposed in this study highlights the interconnected metabolic changes involving mitochondria, peroxisomes, and lysosomes in response to overwork. The findings contribute to our understanding of the mechanisms involved in overwork-related pathologies and provide a basis for further research in the field of karoshi. Conclusion Overall, metabolic reprogramming might provide sufficient energy to the heart, alleviate oxidative stress and damage to cardiac cells in response to excessive exertion and fatigue. Our findings provide an insight into response mechanism to overwork death and lay a foundation for further research on overwork death.

Objective To investigate the occurrence and mechanism of acute Karoshi and explore its forensic identification.Methods SD rats were divided into the control group(n=15)and experimental groups(n=45,acute Karoshi group and overwork survival group).A severe fatigue model was estab-lished by combining forced swimming under load to exhaustion and sleep deprivation.Their daily ac-tivities,diets,weight,respiratory functions,electrocardiogram and echocardiography were recorded.After the rats were sacrificed,samples were collected at autopsies.HE staining was used to observe the pathological morphology,and GC-MS was used to detect the changes of substance metabolism in se-rum,myocardium and liver.Results The mortality rate of the experimental group was 33.3%.There were decreases of aminobutyric acid and arachidonic acid in myocardium tissues,decreases of urea and increases of methionine and phenylalanine in serum.In liver tissues,the content of amino acids sush as histidine increased.The blood biochemical testing showed increases of alanine aminotransferase,as-partate aminotransferase,creatine kinase and creatine kinase isoenzymes and decreases of glucose and uric acid.There were interferences of energy metabolism pathways in serum,heart,and liver tissues.After three days,the experimental group developed cardiac conduction block and ventricular arrhyth-mia.Ventricular fibrillation and ventricular flutter appeared in acute Karoshi group.Echocardiogram showed ejection fraction and left ventricular short axis shortening rate decreased.The histological exami-nation showed granular swelling and sarcoplasmic condensation in myocardium and increased dark neu-rons in the brain stem.The combination of differential metabolites of serum urea,methionine and phe-nylalanine was highly correlated with Karoshi with a diagnostic rate of 90.6%.Conclusion Acute Karoshi can trigger a cascade reaction of metabolic,functional and morphological changes.The mecha-nism of death,especially central failure and sudden cardiac death,may be associated with multi-organ failure.

BACKGROUND:Hepatocellular carcinoma (HCC) stands out as one of the most prevalent malignant tumors globally. The combination of all-trans-retinoic acid (ATRA) with FOLFOX chemotherapy has shown promise in enhancing the prognosis of HCC patients. ATRA, serving as a chemosensitizing agent, presents novel possibilities for therapeutic applications. Nevertheless, the responsiveness of HCC cells to ATRA varies. The epigenetic modifier-GSK-126 is currently under investigation as a potential antitumor drug. Our aim is to explore the molecular mechanisms underlying the diverse sensitivity of HCC patients to ATRA, and to propose a new combination regimen. This research aims to lay the groundwork for personalized medication approaches for individuals with HCC. METHODS:A cell model with low expression of retinoic acid receptor Alfa (RARA), retinoic acid receptor belta (RARB), and retinoic acid receptor gamma (RARG) was established through siRNA interference. The impact of reduced expression of RARA, RARB, and RARG on the half maximal inhibitory concentration (IC50) of ATRA in Hep3B cells was assessed using the 3-(4,5-Dimethyl-2-Thiazolyl)-2,5-Diphenyl Tetrazolium Bromide (MTT) cytotoxicity assay. Flow cytometry revealed that RARG emerged as the key receptor influencing the combination's sensitivity. Conducting ChIP-qPCR analysis on genomic DNA from HCC cells through relevant websites demonstrated enrichment of the trimethylation modification of lysine 27 on histone H3 (H3K27me3) upstream of the RARG promoter. ChIP-PCR assay confirmed that GSK-126 could diminish H3K27me3 levels on the RARG promoter, subsequently elevating RARG expression. The synergistic efficacy of GSK-126 and ATRA was validated through MTT assay, flow cytometry apoptosis assay, cell cycle assay, and cell scratch assay. RESULTS:Our study unveiled that the insensitivity of HCC cells to ATRA could be linked to the low expression of RARG. ChIP-qPCR analysis illuminated that GSK-126 activated RARG expression by diminishing H3K27me3 enrichment in the RARG promoter region. Consequently, the concurrent administration of ATRA and GSK-126 to hepatoma cells exhibited a synergistic effect, inhibiting cell proliferation, inducing cell apoptosis, and reducing the proportion of cells in the S-phase. CONCLUSION:Our findings emphasize that the synergistic action of GSK-126 and ATRA enhances the sensitivity of HCC cells by upregulating the expression of RARG. This presents a potential foundation for personalized HCC treatment.

This work aimed to investigate the pharmacokinetics of six major blood-entering components(triptolide,triptophenolide,wilforgine,wilforine,tripterine,and wilforlide A)in Tripterygium wilfordii Polyglycosides Tablets(TWPT)between normal rats and the rat model of adjuvant arthritis,which was established by injection of Freund's complete adjuvant.Liquid chromatography-mass spectrometry(LC-MS)was employed to determine the content of six main components in the serum samples of normal and arthritic rats after administration of TWPT,and the pharmacokinetics of the six components were compared between normal and model rats.The specificity,linearity,accuracy,precision,stability,extraction recovery and matrix effect of the established LC-MS method all met the requirements.Compared with normal rats,the model rats showed significantly prolonged t1/2 and MRT0-∞ of wilforgine,significantly shortened t1/2 and MRT0-∞ of wilforlide A,and no significant changes in the pharmacokinetics of the other four components in vivo.The results indicated that adjuvant arthritis had different effects on the pharmacokinetics of different active components of TWPT in vivo.The findings provide guidance for the rational clinical application of TWPT and the secondary development of related products.

BACKGROUND:Infertility is a common reproductive disease that affects not only individuals and families, but also the growth of the social population. Hence, understanding the burden of female infertility in China and worldwide is of great significance for the development of infertility prevention and treatment strategies.METHODS:The Global Burden of Disease Study (GBD 2019) Data Resources were used to collect and collate relevant data on female infertility in China and worldwide from 1990 to 2019. The difference in the number, age-standardised prevalence rate (ASPR), disability-adjusted life years and age-standardised disability-adjusted life years rate (ASDR) of women with infertility in different periods and geographical areas were analysed. The autoregressive integrated moving average method was used to predict the ASPR and ASDR of female infertility in China and worldwide in the next 11years.RESULTS:In the past 30years, the number of female infertility cases increased by 7.06million in China and 56.71million worldwide. The corresponding average annual increase of ASPR was 10.10% and 7.28%, respectively, and that of ASDR was 0.08% and 0.79%, respectively. In addition, there are differences in age and time between Chinese and global female infertility. In 1990, the crude prevalence rate of female infertility was the highest in women aged 40-44years and 35-39years in China and worldwide, respectively. In 2019, the crude prevalence rate of female infertility was still the highest in women aged 40-44years in China, whereas that around the world reached the highest in women aged 30-34years, which was significantly earlier. The forecast for the next 11years suggests that the ASPR and ASDR for female infertility in China will first rise and then decline, but the overall magnitude of change is not very significant, whereas the ASPR and ASDR for female infertility globally are still on the rise. The ASPR value of female infertility is expected to be 5025.56 in 100 000 persons in China and 3725.51 in 100 000 persons worldwide by 2030. The ASDR value of female infertility is expected to be 26.16 in 100 000 persons in China and 19.96 in 100 000 persons worldwide by 2030.CONCLUSION:The burden of female infertility is still increasing in China and worldwide. Therefore, it is of great significance to pay more attention to infertile women, and advocate a healthy lifestyle to reduce the burden of disease for infertile women.

目的 探讨自制生化膏在人工流产术后的临床应用价值.方法 选取 2021 年 3 月至 2022 年 7 月于广州市番禺区妇幼保健院门诊实施人工流产手术患者 88 例作为研究对象,按随机数字表法分为对照组与观察组,每组 44 例.对照组口服头孢呋辛酯片,观察组在对照组基础上采用自制生化膏进行治疗,比较两组术后恢复情况.结果 观察组阴道出血时间、月经复潮时间明显短于对照组(P＜0.05);观察组腹痛、首次复潮后月经量减少发生率明显低于对照组(P＜0.05);观察组术后不良反应发生率明显低于对照组(P＜0.05).结论 人工流产术后在应用抗生素的基础上予以自制生化膏,能够减少腹痛发生概率,缩短阴道出血时间和月经复潮时间,降低术后不良反应发生率.

观察EZH2抑制剂(GSK-126)对小鼠肝纤维化的保护作用.将24只实验小鼠随机分为4个小组:对照组(食用油注射量:5 mL/kg)、模型组(CCl4注射量:5 mL/kg的20％的橄榄油溶液)、CCl4+GSK-1265 mg/kg组(CCl4注射量:5 mL/kg的20％的橄榄油溶液,GSK-126注射量:5 mg/kg)、CCl4+GSK-12615 mg/kg组(CCl4注射量:5 mL/kg的20％的橄榄油溶液,GSK-126注射量:15 mg/kg).每3天注射1次,20天后解剖,观察肝脏形态、病理情况及血清ALT值,并通过HE染色和天狼猩红染色来检测GSK-126对小鼠肝纤维化的影响.结果发现:两个GSK-126处理组的肝脏质地较软、肝重指数较轻、ALT活性较低、血管交联和胶原蛋白水平较低,且GSK-12615 mg/kg组比GSK-1265 mg/kg组的肝纤维化程度更低(p<0.05),说明GSK-126对肝纤维化小鼠有明显的保护作用.

目的:探究益经汤加减对早发性卵巢功能不全患者卵巢储备功能的影响.方法:选取2018年2月-2020年6月广州市番禺区妇幼保健院收治的早发性卵巢功能不全患者142例作为研究对象.按照随机数字表法分为常规治疗组和益经汤加减组,每组71例.两组行临床常规治疗,益经汤加减组联合应用益经汤加减治疗.比较两组基础窦卵泡计数、子宫内膜容受性指标[卵巢动脉收缩期峰值流速(PSV)、搏动指数(PI)、阻力指数(RI)、子宫内膜厚度]、性激素[基础促黄体生成素(bLH)、基础雌激素(bE2)、基础促卵泡刺激素(bFSH)]水平、卵巢储备功能[血清抗苗勒管激素(AMH)、抑制素b(INH-b)]、中医症候积分及妊娠率.结果:益经汤加减组治疗后第一个月经期第2～4天的RI低于常规治疗组,基础窦卵泡计数、PSV、PI、子宫内膜厚度均高于常规治疗组,差异均有统计学意义(P<0.05).益经汤加减组治疗后第一个月经期结束bLH、bFSH水平均低于常规治疗组,bE2水平高于常规治疗组,差异均有统计学意义(P<0.05).益经汤加减组治疗后第一个月经期第2～3天AMH、INH-b水平均高于常规治疗组(P<0.05).益经汤加减组治疗后中医症候主症、次症评分和总分均低于常规治疗组(P<0.05).益经汤加减组妊娠率高于常规治疗组(P<0.05).结论:益经汤加减可显著改善早发性卵巢功能不全患者性激素、卵巢储备功能,改善INH-b、bLH异常表达,促进卵巢功能恢复,提高妊娠率.

Polycystic ovary syndrome (PCOS) is a common endocrine disease in women, potentially causing ovarian infertility for women at gestational age. Huatan Tongjing Decoction is commonly used to treat PCOS; however, the involved molecular mechanism has not been fully understood. In this study, the active components of Huatan Tongjing Decoction and potentially targeted proteins were downloaded from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. PCOS-related genes were accessed from Malacards database. STRING database was utilized to construct a protein-protein interaction (PPI) network based on the PCOS-related genes and the predicted targets. Subsequently, the PPI network was subjected to Random walk with restart (RWR). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on top 50 genes with the high affinity scores to the drug targets. Subsequently, based on the predicted drug components and targets, a component-gene interaction network was constructed. Finally, the most central drug targets were selected, and the corresponding compounds were subjected to molecular docking and dynamic simulations to examine their bindings. The 122 main active components and 246 potential targets of Huatan Tongjing Decoction were obtained from TCMSP, and a total of 259 nodes and 1919 interactions were acquired from the PPI network. The top 50 genes were mainly enriched in response to peptide hormone function and PI3K-Akt signaling pathway. Molecular docking and dynamic simulations predicted that MMP-quercetin interaction played an important role in the treatment of PCOS using Huatan Tongjing Decoction. Luteolin and quercetin in Huatan Tongjing Decoction potentially bound MMP9 and served as active components. This study preliminarily suggested the efficacy of Huatan Tongjing Decoction against PCOS in molecular degree.

目的:探究温针灸结合来曲唑治疗多囊卵巢综合征不孕肾虚血瘀型的临床疗效.方法:100例分成两组各50例,两组均用来曲唑治疗,观察组加用温针灸治疗.结果:治疗后观察组中医证候总积分低于对照组(P＜0.05),观察组各项性激素指标数值均优于对照组(P＜0.05),观察组卵泡直径和子宫内膜厚度均优于对照组(P＜0.05),观察组妊娠率高于对照组(P＜0.05),观察组生活质量评分高于对照组.结论:温针灸结合来曲唑治疗多囊卵巢综合征不孕肾虚血瘀型效果较好.

中性粒细胞胞外诱捕网是活化的中性粒细胞分泌的由去聚化染色质DNA骨架和多种抗菌蛋白组成的细胞外纤维网状结构，通过诱捕病原菌参与机体的免疫应答。近年来，多项研究表明，中性粒细胞胞外诱捕网在肿瘤转移中发挥重要的作用。本文主要关注中性粒细胞胞外诱捕网形成的机制、在肿瘤转移中的作用以及在判断肿瘤预后和抗肿瘤治疗中的潜在作用。

[目的]细菌机械敏感性离子通道MscS能够在细菌周围环境渗透压急剧降低时,打开并释放胞内内容物,平衡内外渗透压差,使细菌存活.鉴于其广泛分布在各种细菌中,而在哺乳动物中未发现其同源体,MscS被认为是一种新型抗生素靶点.MscS一个独特的开放特征是具有失活特性,即在持续的机械刺激条件下,MscS从开放状态进入一种非离子通透的失活状态,从而避免因通道持续开放引起大量内容物流失导致细菌死亡.该研究的目的是鉴定影响MscS失活的关键氨基酸,为靶向MscS的药物设计提供思路.[方法]采用分子克隆方法制备MscS Cyto-helix(P166-I170)半胱氨酸突变体,利用巯基化合物MTSET+结合半胱氨酸从而对其侧链基团进行修饰,并通过低渗刺激实验,检测表达MscS半胱氨酸突变体的大肠杆菌分别在无或有MTSET+处理下,低渗刺激诱发通道开放后的存活率筛选显著影响通道功能的突变体.利用电生理膜片钳方法检测突变体在MTSET+处理前后通道失活特性的变化,结合定点突变手段进一步探讨失活机制.[结果]MTSET+处理导致表达半胱氨酸突变体G168C-MscS的大肠杆菌在低渗刺激后存活率极大降低;G168C-MscS在结合MTSET+后失去失活特性,保持持续开放,是导致细菌胞内内容物大量流失并死亡的重要原因;酪氨酸突变G168Y-MscS、亮氨酸突变G168L-MscS和赖氨酸突变G168K-MscS的失活特性与野生型WT-MscS 一致,而天冬氨酸突变G168D、缬氨酸突变G168V和异亮氨酸突变G168I的失活速率显著降低,尤其是G1681-MscS失去失活特性,表明MscS 168位点是影响通道失活的关键位点,并且通道失活特性与该位点氨基酸侧链基团的大小及电荷性质相关.[结论]G168位点甘氨酸是影响MscS通道失活的关键氨基酸.

Shengmai injection (SMI), a traditional Chinese medicine formula with the nature of multicomponent and multi-target, has been widely used in clinic for treating cardiovascular diseases in China; however, its comprehensive mechanism of action remains unclear. In this study, a TMT-based quantitative serum proteomics was performed to explore SMI's global mechanism and help identify serum biomarkers of its effect on isoproterenol (ISO)-induced myocardial ischemia rats. The results of TMT-based proteomic analysis identified 227, 100, and 228 differentially expressed proteins (DEPs) for the model compared to the control group, SMI pretreatment + model compared to the model group, and SMI pretreatment + model compared to the control group, respectively. Based on bioinformatics analyses of gene ontology (GO), KEGG pathways, and the protein-protein interaction (PPI) networks for the DEPs, it is concluded that the comprehensive mechanism of SMI's effect on ISO-induced myocardial ischemia injury includes regulation of energy metabolism, reducing endothelial cell permeability, regulation of vessel and cardiac contractility, anti-inflammation, and prevention of cell apoptosis. Furthermore, 10 common DEPs were found, and six of them were regulated in model vs. control group, while back-regulated in SMI pretreatment + model vs. model group. Among them, three functional proteins of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Fas apoptotic inhibitory molecule 3 (FAIM3), and uncharacterized protein (M0R5J4), which were verified by the PRM analysis, might be the potential serum biomarkers on SMI's effects. Overall, this serum proteomics of SMI not only provides insights into the comprehensive mechanism underlying SMI's effects on ischemic heart disease but also helps identify serum biomarkers for directing SMI's cardioprotective effects.

肝纤维化是常见的慢性进行性肝病,是慢性肝病发展到肝硬化阶段的必经阶段,却有逆转的可能性.肝纤维化的中心环节是肝星状细胞的激活.关于肝星状细胞的激活,除了经典的肝纤维化通路,不断有新的通路和机制出现,包括自噬、内质网应激、氧化应激、胆固醇代谢和表观遗传,这些都揭示了肝星状细胞的活化机制.表观遗传包括DNA甲基化、组蛋白修饰和调节性非编码RNAs,这些机制也参与调节肝星状细胞活化和肝纤维化发生,对表观遗传和肝纤维化治疗之间的关系研究具有重要意义.

生脉注射液(Shengmai Injection,SMI)是由红参、麦冬和五味子3味中药组成的中药复方制剂,在临床上除了广泛用于治疗心血管系统疾病外,还用于抗肺病、癌症化疗等的辅助治疗.近年来,SMI的临床前基础研究日渐广泛,主要包括SMI的化学成分、体内过程和作用机制等,为揭示该方的药效物质基础提供了一定的科学依据.综述了该方的物质组成、临床前药动学和药效学研究,以期为SMI的质量控制、产品二次开发和临床合理应用提供参考.

Background/Aims: Liver progenitor cells (LPCs) were considered as a promising hepatocyte source of cell therapy for liver disease due to their self-renewal and differentiation capacities, while little is known about the mechanism of LPC differentiate into hepatocytes. This study aims to explore the effect of miR-382, a member of Dlk1-Dio3 microRNA cluster, during hepatic differentiation from LPCs. Methods: In this study, we used rat liver progenitor cell WB-F344 as LPC cell model and HGF as inducer to simulate the process of LPCs hepatic differentiation, then microRNAs were quantified by qPCR. Next, WB-F344 cell was transfected with miR-382 mimics, then hepatocyte cell trait was characterized by multiple experiments, including that periodic acid schiff staining and cellular uptake and excretion of indocyanine green to evaluate the hepatocellular function, qPCR and Western Blotting analysis to detect the hepatocyte-specific markers (ALB, Ttr, Apo E and AFP) and transmission electron microscopy to observe the hepatocellular morphology. Moreover, Luciferase reporter assay was used to determine whether Ezh2 is the direct target of miR-382. Results: We found that miR-382 increased gradually and was inversely correlated with the potential target, Ezh2, during WB-F344 hepatic differentiation. In addition, functional studies indicated that miR-382 increased the level of hepatocyte-specific genes. Conclusions: This study demonstrates that miR-382 may be a novel regulator of LPCs differentiation by targeting Ezh2.

The incidence and mortality of liver disease are very high. So far, liver transplantation remains the only effective treatment for end-stage liver failure but is limited by the availability of donor organs. After decades of development, cell therapy in people has taken the form of hepatocyte transplantation for metabolic disorders with a hepatic basis, and for acute or chronic liver failure. Compared to orthotopic liver transplantation,hepatocyte transplantation is featured with the advantages of sufficient resource, small trauma, easy to operate and affordable, so it plays an important role in the treatment of endstage liver disease and metabolic liver disease. However,hepatocyte transplantation still has some disadvantages, such as low activity, small number of donor cells,easy rejection and lack of proliferation after transplantation, restricting the development of hepatocyte transplantation.

目的 基于二硫键交联合成精氨酸-组氨酸(HRss)阳离子聚合物,构建新型纳米复合物HRss/miR-15-a,用于前列腺癌研究.方法 利用1 H-NMR鉴定HRss2/miR-15-a载体的结构,通过电位粒度仪测定纳米复合物的电位和粒径,利用琼脂糖凝胶电泳考察载体对miR-15-a的包裹能力.以体外培养的前列腺癌干细胞RC-92a/hTERT为研究对象,CCK8法检测3种纳米复合物对细胞增殖的抑制作用,以pEGFP为报告基因考察基因转染效率,利用Transwell小室考察HRss/miR-15-a对细胞运动能力的影响.结果 细胞毒性试验结果显示,载体材料对正常及肿瘤细胞毒性较低,可以包裹miR-15-a形成稳定的纳米复合物,HRss2/miR-15-a相较于HRss1/miR-15-a和HRss3/miR-15-a,转染效率更高,对前列腺癌干细胞的运动能力影响也更大.结论 HRss可以运载基因药物,体外评价效果最佳的HRss2,有潜力成为抗前列腺癌基因治疗中的新型载体.

Brain ischemia/reperfusion (I/R) injury (BI/RI) is a leading cause of death and disability worldwide. However, the outcome of pharmacotherapy for BI/RI remains unsatisfactory. Innovative approaches for enhancing drug sensitivity and recovering neuronal activity in BI/RI treatment are urgently needed. The purpose of this study was to evaluate the protective effects of tumor necrosis factor (TNF)-α-loaded poly(ethylene glycol)-b-(poly(ethylenediamine L-glutamate)-g-poly(L-lysine)) (TNF-α/PEG-b-(PELG-g-PLL)) nanoparticles on BI/RI. The particle size of PEG-b-(PELG-g-PLL) and the loading and release rates of TNF-α were determined. The nanoparticle cytotoxicity was evaluated in vitro using rat cortical neurons. Sprague Dawley rats were preconditioned with free TNF-α or TNF-α/PEG-b-(PELG-g-PLL) polyplexes and then subjected to 2 hours ischemia and 22 hours reperfusion. Brain edema was assessed using the brain edema ratio, and the antioxidative activity was assessed by measuring the superoxide dismutase (SOD) activity and the malondialdehyde (MDA) content in the brain tissue. We further estimated the inflammatory activity and apoptosis level by determining the levels of interleukin-4 (IL-4), IL-6, IL-8, IL-10, and nitric oxide (NO), as well as the expression of glial fibrillary acidic protein (GFAP), intercellular adhesion molecule-1 (ICAM-1), and cysteine aspartase-3 (caspase-3), in the brain tissue. We provide evidence that TNF-α preconditioning attenuated the oxidative stress injury, the inflammatory activity, and the apoptosis level in I/R-induced cerebral injury, while the application of block copolymer PEG-b-(PELG-g-PLL) as a potential TNF-α nanocarrier with sustained release significantly enhanced the bioavailability of TNF-α. We propose that the block copolymer PEG-b-(PELG-g-PLL) may function as a potent nanocarrier for augmenting BI/RI pharmacotherapy, with unprecedented clinical benefits. Further studies are needed to better clarify the underlying mechanisms.