Hemorrhagic fever with renal syndrome virus infection in livers studied by in situ hybridization and immunohistochemistry

ＨｅｍｏｒｒｈａｇｉｃｆｅｖｅｒｗｉｔｈｒｅｎａｌｓｙｎｄｒｏｍｅｖｉｒｕｓｉｎｆｅｃｔｉｏｎｉｎｌｉｖｅｒｓｓｔｕｄｉｅｄｂｙｉｎｓｉｔｕｈｙｂｒｉｄｉｚａｔｉｏｎａｎｄｉｍｍｕｎｏｈｉｓｔｏｃｈｅｍｉｓｔｒｙＹａｎｇＳｈｏｕｊｉｎｇ（杨守京）；Ｌ...     

2型糖尿病患者外周血白细胞iNOSmRNA表达的变化及意义

目的研究2型糖尿病DM患者外周血白细胞中iNOSmRNA表达的变化及其与糖尿病肾病DN发生、发展的关系。方法101例2型DM患者,根据尿微量白蛋白排泄率和血肌酐水平分为单纯DM组和不同的DN组,用原位杂交法检测外周血白细胞iNOSmRNA表达的阳性细胞的百分率,并与21例健康体检者进行比较。结果早期DN组白细胞iNOSmRNA表达的百分率明显高于对照组、DM组及晚期DN组(P<0.001)。结论外周血白细胞iNOSmRNA表达的变化参与了DN的发生、发展。     

Protective effect of interferon-α on the DNA- and RNA-degrading pathway in anti-Fas-antibody induced apoptosis

Aim: Fas membrane-associated polypeptide antigen is a receptor molecule responsible for apoptosis-mediated signals. In animal models of acute viral hepatitis, apoptosis of hepatocytes is mediated by Fas-death receptors; therefore, the aim of this study was to evaluate the effect of interferon (IFN)-alpha on apoptotic markers and nuclease activity against different coding and non-coding single and double stranded RNAs during Fas-induced liver apoptosis. Methods: An in vivo experiment was performed with simultaneous administration of anti-Fas (CD95) antibodies and IFN-alpha, and an in vitro experiment was performed in hepatocyte cultures treated with anti-Fas antibodies and IFN-alpha. Results: Detection of apoptosis using Annexin V-FITC/propidium iodide, Bcl-2 and Bax expression in hepatocyte cultures confirmed the appearance of early apoptotic events and progression toward late apoptosis after anti-Fas antibody treatment. IFN-alpha had a tendency to retard the apoptosis process in Fas-induced apoptosis by increasing the number of viable cells and decreasing the number of cells in late apoptosis, by increasing the percentage of Bcl-2 positive cells, by decreasing the percentage of Bax positive cells, and by decreasing the nuclease activity compared to the anti-Fas antibody treated group. Total DNA and RNA concentration was much reduced in the Fas group and DNA fragmentation assay provided evidence for increased DNA degradation. Enhanced nuclease activity against DNA, rRNA, poly(A), poly(C), poly(U), poly(I:C), and poly(A:U) was manifested in the anti-Fas antibody treated group, except for the inhibitory-bound alkaline RNase. Conclusions: The results demonstrate that the RNA-degrading pathway in Fas-induced apoptosis can accelerate the liberation of the latent enzyme from the inhibitor complex. IFN-alpha prevented enormous, Fas-ligand induced degradation of all the substrates used in this experimental study, most probably due to similarities in the signal transduction pathways. Investigations of death receptor-induced apoptosis may lead to novel treatment combinations for patients with acute or chronic liver diseases.     

乙型肝炎病毒转基因小鼠体内肝靶向反义RNA抗病毒疗效研究

目的：探讨半乳糖修饰的反义RNA真核表达质粒在乙型肝炎病毒转基因小鼠体内的抗病毒作用。方法：以半乳糖多聚赖氨酸（Gal-PLL)作肝靶向载体，将乙型肝炎病毒基因C区的反义RNA真核表达重组质粒（pCEP4-aC）制备为Gal-PLL－pCEP4-aC。将24只血清HBV DNA、HBsAg阳性的小鼠，随机等分为Gal-PLL－pCEP4-aC治疗组、Gal-PLL-pCEP4对照组和生理盐水阴性对照组，于实验第1天尾静脉分别注射Gal-PLL－pCEP4-aC、Gal-PLL－pCEP4（100μg/只）和等体积的生理盐水，观察治疗前后血清HBV DNA以及HBsAg变化。结果：Gal-PLL－pCEP4-aC治疗组21天时血清HBV－DNA转阴率62．5％（5／8），且7，14，21天时血清HBsAg明显降低；而Gal-PLL－pCEP4组血清HBV DNA转阴1只（1／8），生理盐水组8只均未转阴，两组用药后血清中HBsAg与用药前比较差异性均不明显（P＞0．05）。结论：肝靶向反义RNA能在乙肝基因小鼠体内抑制HBV的复制和抗原表达。     

Prevalence of anti-HCV in institutionalised mentally handicapped subjects

Data on the prevalence of antibody to hepatitis C virus among 278 subjects in an institution for the mentally handicapped were analysed by risk factors. A prevalence of 4% was found, higher than blood donors belonging to the same area (1.4%). No differences in prevalence with regard the length of residence, age, sex, degree of retardation or for presence of HBV markers were observed.     

补中益气汤治疗慢性乙型肝炎的临床与实验研究

通过对补中益气汤及西药治疗慢性乙型肝炎临床疗效的对比研究,发现补中益气汤能较快改善慢性乙型肝炎的临床症状及体征,改善肝功能,促使乙型肝炎病毒血清学标志好转(HBsAg、HBeAg转阴或滴度下降,抗-HBe或抗-HBs转为阳性),与西药治疗对照组比较有显著性差异(P<0.05)。动物实验观察了补中益气汤对小鼠肝组织DNA、RNA、蛋白质合成的影响,结果表明,补中益气汤对三者的合成均有明显的促进作用。其抗肝炎的作用机制可能与该方增强肝脏蛋白质的合成,促进肝组织的修复,改善机体整体的抗病机能有关。     

Cdh1小干扰RNA载体的构建及鉴定

目的:构建Cdh1小干扰RNA载体,并将其转染至Hela细胞进行鉴定.方法:根据pENTRTM/H1/TO中间载体要求,设计Cdh1小干扰RNA载体的干扰序列及无干扰作用的对照序列,合成相应的DNA单链,退火后连接到pENTRTM/H1/TO线性载体,形成完整载体,进行测序鉴定.分别将测序鉴定成功的Cdh1小干扰RNA载体及对照载体采用脂质体法转染Hela细胞,转染后48 h提取细胞总RNA及细胞总蛋白,采用实时定量PCR和Western Blot检测Cdh1的表达.结果:经测序鉴定成功构建Cdh1小干扰RNA载体和对照载体,分别命名为pENTR/shCdh1和pENTR/shcontrol.与未转染及转染pENTR/shcontrol的Hela细胞相比,转染pENTR/shCdh1的Hela细胞Cdh1表达降低(P《0.05).结论:成功构建Cdh1小干扰RNA载体,并能下调Hela细胞Cdh1的表达.