慢性肝炎中乙型和丙型病毒混合感染的重症化研究

在慢性肝炎中,乙、丙型病毒性肝炎混合感染相当多见,可使肝炎慢性化、重症化,肝组织损伤加重,肝硬化(LC)和肝癌(HCC)发生率增加[1]。本文应用血清学和分子生物学方法对196例肝病患者的血清进行检测,初步探讨了乙型肝炎病毒(Hepatitis B virus,HBV)、丙型肝类病毒(Hepatitis C virus,HCV)的复制状况以及两者间的相互作用与预后的关系。1材料和方法1.1病例受检的196例病例均为2004年1月至2005年7月我院住院及门诊病人,男149例,女47例,年龄15~82岁,其中慢性肝炎(CH)患者139例,肝硬化(LC)患者42例,肝癌(HCC)患者15例。所有病例诊断符合…     

两例丙型肝炎病毒(HCV)和乙型肝炎病毒(HBV)重叠感染者的HCV核心区cDNA序列分析

从临床肝病患者中选择两例ＨＣＶ和ＨＢＶ重叠感染者ＨＳＱ和ＳＺＨ,他们血清中的生化指标丙氨酸转氨酶（ＡＬＴ）持续异常,肝活检病理示有严重的肝损伤。在ＡＬＴ异常期,血清学检测结果为ＨＢｓＡｇ、ＨＢｅＡｇ阳性,抗ＨＣＶＩｇＧ（包括Ｃ２２、Ｃ３３ｃ）阴性,但套式ＰＣＲ检测ＨＣＶＲＮＡ阳性,核心区ｃＤＮＡ序列分析发现该区有１个密码子（ＧＧＣｎｔ３８５—３８７）缺失,对应缺失的氨基酸是甘氨酸（ＧＬＹ）,从血清学检测和序列分析结果推测,在ＨＣＶ和ＨＢＶ重叠感染中,ＨＢＶ和ＨＣＶ均可处于持续复制状态,抗ＨＣＶＩｇＧ抗体阴性可能是ＨＣＶ的多蛋白前体翻译和病毒颗粒装配受到ＨＢＶ干扰的结果。     

丙型肝炎病毒的感染特点

１９８８年非甲非乙型肝炎病毒被命名为丙型肝炎病毒（ＨＣＶ）,随后建立了血清学诊断方法和更特异的免疫杂交和多聚酶链反应（ＰＣＲ）法。感染ＨＣＶ后病人多呈慢性过程,可导致肝硬化及肝癌。ＨＣＶ主要经胃肠道外传播,静脉药瘾者为高危人群,性传播、家庭内密切接触和母婴传播不占重要地位。治疗可用ＩＦＮ－α,但易复发。ＨＣＶ感染后免疫保护作用差,尚无疫苗可用。     

丙型肝炎病毒性传播感染研究进展

随着性传播疾病（ＳＴＤ）的流行,丙型肝炎病毒（ＨＣＶ）性传播感染日益受到重视。本文综合分析ＨＣＶ性传播感染流行病学调查研究的不同观点,旨在提醒人们对ＨＣＶ性传播感染的警惕。     

丙型肝炎病毒核心基因置换对病毒复制和感染影响的初步研究

本文旨在探讨丙型肝炎病毒(HCV)核心(core)蛋白在病毒复制和感染中的作用,采取基因置换的方法,用HCV1b型J4株的核心基因平行置换2a型J6JFH1株的核心区,构建了FL-J6JFH/J4core嵌合复制子。体外制备RNA转录体,以脂质体介导转染Huh7.5.1肝癌细胞。转染后第5天,用荧光定量聚合酶链反应(FQ-PCR)检测细胞内HCVRNA水平。第8天,以丙型肝炎患者血清为一抗,免疫荧光法(IFA)检测转染细胞内HCV蛋白表达。同时收集转染后第8天的细胞上清液,感染naveHuh7.5.1肝癌细胞,72h后IFA检测HCV蛋白表达。结果显示,FL-J6JFH/J4core转染组第5天细胞内RNA水平与野生型FL-J6JFH1接近,无显著差异(n=4,P0.05)。免疫荧光检测显示,FL-J6JFH/J4core转染细胞后第8天及其上清液感染的naveHuh7.5.1细胞的HCV阳性细胞数都低于野生型。本研究结果初步表明,HCV各株间核心基因的替换会影响HCV的蛋白翻译和感染性病毒颗粒的产生与释放。     

人类免疫缺陷病毒/丙型肝炎病毒混合感染者免疫细胞和肝生化指标的分析

目的 探讨人类免疫缺陷病毒/丙型肝炎病毒(HIV/HCV)混合感染者的免疫淋巴细胞亚群和肝脏生化指标的特征。方法 检测并分析研究组(40例混合感染HIV/HCV的血友病患者)和对照组(12例单独感染HIV的血友病患者)若干相关实验室指标。结果两组(研究与对照)结果 差异显著的指标分别为:自然杀伤(NK)细胞9.26%±5.98%对12.19%±5.80%,P<0.05;CD4/CD8细胞比值0.39±0.21对0.53±0.24,P<0.05;CD3细胞74.99%±10.33%对68.42%±8.82%,P<0.05;CD8细胞52.28%±12.59%对43.58%±10.99%,P<0.05;丙氨酸氨酶(ALT)(48.59±40.21)U/L对(26.87±27.23)U/L,P<0.01;天冬氨酸转氨酶(AST)(61.66±51.02)U/L对(34.17±30.24)U/L,P<0.001;谷氨酰转肽酶(GGT)(136.50±111.50)U/L对(43.88±36.17)U/L,P<0.001;甘胆酸(CG)(683.41±962.22)μg/L对(250.96±290.81)μg/L,P<0.001;透明质酸(HA)(180.94±196.69)ng/ml对(64.68±32.74)ng/ml,P<0.001;白/球蛋白(A/G)比值1.35±0.24对1.50±0.34,P<0.05。与单独HIV感染者比较,HIV/HCV混合感染者的NK细胞、CD4/CD8细胞比值和A/G比值显著降低,而ALT、AST、GGT、CG和HA的水平显著增加。结论 HIV/HCV混合感染可能加剧患者淋巴细胞亚群的不平衡,加重肝脏损伤和出现肝硬化趋势。     

丙型肝炎病毒核心蛋白抑制乙型肝炎病毒转录的分子机理

为研究丙型肝炎病毒 (HCV)核心蛋白抑制乙型肝炎病毒 (HBV)表达的分子机理 ,从HCV和HBV共感染中国病人血清中分离了 5个含HCV 5′非编码区 ( 5′NCR)和核心区 (CR)cDNA序列的克隆 .序列比较和系统发育分析显示 :从共感染病例中分离的HCV序列与其它已发表的从单独HCV感染病例中分离的序列没有显著差异 .共转染实验证实采用从其中 1例共感染病人中分离的核心蛋白cDNA基因所表达的核心蛋白 ,可抑制HBV表面抗原 (HBsAg)和HBVe抗原 (HBeAg)的表达 ,缺失突变分析说明HCV核心蛋白的C端疏水区对这种抑制作用是必需的 ,报道基因产物分析进一步说明了HBVC启动子和增强子Ⅱ是HCV核心蛋白的效应靶序列之一 ,而HCV核心蛋白在肝细胞和非肝细胞中均对HBV和其他细胞和病毒的基因的转录起负调节作用 .因此 ,认为HCV核心蛋白是一种多功能的负调节因子 ,与HCV和HBV以及HCV与细胞之间的相互作用密切相关     

肝硬变中丙型肝炎病毒C33c抗原及HBxAg的分布及意义

应用抗ＨＣＶＣ３３ｃ抗原２Ｂ６株单克隆抗体和抗ＨＢｘＡｇ多克隆抗体。以ＡＢＣ法对８６例肝硬变组织进行ＨＣＶ及ＨＢＶ相关抗原定位研究,ＨＣＶＣ３３ｃ抗原及ＨＢｘＡｇ在肝硬变中的阳性率分别为７６．７％及６２．８％,Ｃ３３ｃ抗原和ＨＢｘＡｇ阳性占所检病例８８．４％,二者同时阳性为５１．２％,ＨＣＶＣ３３ｃ抗原位于肝硬变组织的肝细胞胞桨内,充满整个胞桨,细胞核及胸膜未见阳性;阳性细胞呈弥温、局灶及散在分布     

HIV and HCV: from Co-infection to epidemiology, transmission, pathogenesis, and treatment

Human immunodeficiency virus (HIV) is the infectious agent causing acquired immunodeficiency syndrome (AIDS), a deadliest scourge of human society. Hepatitis C virus (HCV) is a major causative agent of chronic liver disease and infects an estimated 170 million people worldwide, resulting in a serious public health burden. Due to shared routes of transmission, co-infection with HIV and HCV has become common among individuals who had high risks of blood exposures. Among hemophiliacs the co-infection rate accounts for 85%; while among injection drug users (IDU) the rate can be as high as 90%. HIV can accelerate the progression of HCV-related liver disease, particularly when immunodeficiency has developed. Although the effect of HCV on HIV infection is controversial, most studies showed an increase in mortality due to liver disease. HCV may act as a direct cofactor to fasten the progression of AIDS and decrease the tolerance of highly active antiretroviral therapy (HARRT). Conversely, HAART-related hepatotoxicity may enhance the progression of liver fibrosis. Due to above complications, co-infection with HCV and HIV-1 has imposed a critical challenge in the management of these patients. In this review, we focus on the epidemiology and transmission of HIV and HCV, the impact of the two viruses on each other, and their treatment.      

HIV and HCV： from Co-infection to Epidemiology, Transmission, Pathogenesis, and Treatment

Human immunodeficiency virus (HIV) is the infectious agent causing acquired immu-nodeficiency syndrome (AIDS),a deadliest scourge of human society. Hepatitis C virus (HCV) is a major causative agent of chronic liver disease and infects an estimated 170 million people worldwide,resulting in a serious public health burden. Due to shared routes of transmission,co-infection with HIV and HCV has become common among individuals who had high risks of blood exposures. Among hemophiliacs the co-infection rate accounts for 85%; while among injection drug users (IDU) the rate can be as high as 90%. HIV can accelerate the progression of HCV-related liver disease,particularly when immunodeficiency has developed. Although the effect of HCV on HIV infection is controversial,most studies showed an increase in mortality due to liver disease. HCV may act as a direct cofactor to fasten the progression of AIDS and decrease the tolerance of highly active antiretroviral therapy (HARRT). Conversely,HAART-related hepatotoxicity may enhance the progression of liver fibrosis. Due to above complications,co-infection with HCV and HIV-1 has imposed a critical challenge in the management of these patients. In this review,we focus on the epidemiology and transmission of HIV and HCV,the impact of the two viruses on each other,and their treatment.     

结合RT-PCR体外扩增检测HCV ssRNA的研究

本研究旨在以HCV为平台,在简化RT-PCR基础上,结合体外转录,建立一种特异、高效、简便的检测血清中HCV RNA的体外转录合成系统.本法扩增终产物为特定极性的ssRNA,其特异性经凝胶电泳和斑点杂交确定;RNA定量分析结果显示本法核酸扩增效率高于巢式PCR近20倍.     

NS5A protein of HCV enhances HBV replication and resistance to interferon response

Patients with acute vitiligo have low epidermal catalase expression/activities and accumulate 10(-3) M H(2)O(2). One consequence of this severe oxidative stress is an altered calcium homeostasis in epidermal keratinocytes and melanocytes. Here, we show decreased epidermal calmodulin expression in acute vitiligo. Since 10(-3)M H(2)O(2) oxidises methionine and tryptophan residues in proteins, we examined calcium binding to calmodulin in the presence and absence of H(2)O(2) utilising (45)calcium. The results showed that all four calcium atoms exchanged per molecule of calmodulin. Since oxidised calmodulin looses its ability to activate calcium ATPase, enzyme activities were followed in full skin biopsies from lesional skin of patients with acute vitiligo (n=6) and healthy controls (n=6). The results yielded a 4-fold decrease of ATPase activities in the patients. Computer simulation of native and oxidised calmodulin confirmed the loss of all four calcium ions from their specific EF-hand domains. Taken together H(2)O(2)-mediated oxidation affects calcium binding in calmodulin leading to perturbed calcium homeostasis and perturbed l-phenylalanine-uptake in the epidermis of acute vitiligo.     

黑龙江省乙型肝炎血清流行病学调查分析

为了掌握黑龙江省乙型肝炎(乙肝)病毒(HBV)流行现状和人群免疫水平,评价我省儿童乙肝疫苗预防接种效果,为制订乙肝预防控制策略提供依据。采用横断面调查方法,选择全省7个国家级疾病监测点,在每个监测点随机抽取2个乡级单位,1～59岁作为目标人群共调查3337名人群。黑龙江省1～59岁人群乙肝病毒表面抗原(HBsAg)阳性率、乙肝病毒表面抗体(抗-HBs)阳性率和乙肝病毒核心抗体(抗-HBc)阳性率经标化后为5.65%、55.45%、30.50%,1～3岁人群乙肝表面抗原阳性率明显低于15～59岁人群。1～3岁、4～10岁和11～14岁人群乙肝疫苗全程接种率为99.43%、89.40%和64.81%;首针及时接种率分别为86.64%、75.57%和49.87。城市和农村HbsAg阳性率分别为3.26%和5.04%。医院出生儿童乙肝疫苗首针及时接种率高于在家出生儿童。结果显示接种乙肝疫苗可明显提高抗-HBs阳性率,提高人群对HBV的免疫保护能力,降低HBsAg携带率。     

结合RT-PCR体外扩增检测HCVssRNA的研究

本研究旨在以HCV为平台,在简化RT-PCR基础上,结合体外转录,建立一种特异、高效、简便的检测血清中HCVRNA的体外转录合成系统。本法扩增终产物为特定极性的ssRNA,其特异性经凝胶电泳和斑点杂交确定;RNA定量分析结果显示本法核酸扩增效率高于巢式PCR近20倍。     

HCV核心蛋白诱导Cos—7细胞凋亡

将丙型肝炎病毒(HCV)核心蛋白(Core)基因的全长序列插入到真核表达载体pCDNA3 CMV启动子下游,构建真核表达载体pCDNA3—Core,用脂质体LipoVec^TM转染Cos—7细胞系进行瞬时表达;DNA转染24h后用免疫斑点试验检测在细胞中表达的Core蛋白;转染72h后用Hoechst染色和DNA Ladder检测Cos—7细胞的凋亡情况;荧光染色观察到了细胞凋亡核碎裂,琼脂糖凝胶电泳也呈现出180—200bp整数倍的梯形带,呈现典型的细胞凋亡特征。这些结果表明HCV Core蛋白的表达能引起Cos—7细胞凋亡,Core蛋白的这种功能可能在HCV的持续感染过程中起着一定的作用。     

HCV核心蛋白诱导Cos-7细胞凋亡

将丙型肝炎病毒(HCV)核心蛋白(Core)基因的全长序列插入到真核表达载体pCDNA3 CMV启动子下游,构建真核表达载体pCDNA3-Core,用脂质体LipoVecTM转染Cos-7细胞系进行瞬时表达;DNA转染24h后用免疫斑点试验检测在细胞中表达的Core蛋白;转染72h后用Hoechst染色和DNA Ladder检测Cos-7细胞的凋亡情况;荧光染色观察到了细胞凋亡核碎裂,琼脂糖凝胶电泳也呈现出180-200bp整数倍的梯形带,呈现典型的细胞凋亡特征.这些结果表明HCV Core蛋白的表达能引起Cos-7细胞凋亡,Core蛋白的这种功能可能在HCV的持续感染过程中起着一定的作用.