庚型病毒性肝炎22例临床观察(附7例病理报告)

目的：观察庚型肝炎（ Ｈ Ｇ）临床、生化和病理特点。方法：用逆转录聚合酶链反应（ Ｒ Ｔ Ｐ Ｃ Ｒ）检测血清 Ｈ Ｇ Ｖ Ｒ Ｎ Ａ,并对２２ 例 Ｈ Ｇ的临床表现、肝功能检查和７ 例肝组织的病理特征进行分析。结果：（１） Ｈ Ｇ 的传播以输血及血制品和注射途径为主;（２）７ 例单纯 Ｈ Ｇ 的临床表现有一定隐匿性,血清丙氨酸转氨酶（ Ａ Ｌ Ｔ）、总胆红素（ Ｔｂｉｌ）、白蛋白（ Ａ）、 Ａ／ Ｇ、和凝血酶原活动度（ Ｐ Ｔ Ａ）的异常程度均明显轻于 Ｈ Ｇ重叠慢丙肝（９ 例）或慢乙肝者（６ 例）（ Ｐ＜ ０．０５）;（３）肝组织学示３ 例单纯 Ｈ Ｇ者呈急性轻型肝炎改变,而４ 例 Ｈ Ｇ 合并慢丙肝或慢乙肝者的肝组织则呈现不同程度的炎症活动和肝纤维化。结论：单纯 Ｈ Ｇ Ｖ 感染的临床症状和肝损害均较轻、预后良好,而 Ｈ Ｇ重叠慢性乙肝或慢性丙肝者则肝功能损害可较明显。     

An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis

A specific assay has been developed for a blood-borne non-A, non-B hepatitis (NANBH) virus in which a polypeptide synthesized in recombinant yeast clones of the hepatitis C virus (HCV) is used to capture circulating viral antibodies. HCV antibodies were detected in six of seven human sera that were shown previously to transmit NANBH to chimpanzees. Assays of ten blood transfusions in the United States that resulted in chronic NANBH revealed that there was at least one positive blood donor in nine of these cases and that all ten recipients seroconverted during their illnesses. About 80 percent of chronic, post-transfusion NANBH (PT-NANBH) patients from Italy and Japan had circulating HCV antibody; a much lower frequency (15 percent) was observed in acute, resolving infections. In addition, 58 percent of NANBH patients from the United States with no identifiable source of parenteral exposure to the virus were also positive for HCV antibody. These data indicate that HCV is a major cause of NANBH throughout the world.     

Complete replication of hepatitis C virus in cell culture

Many aspects of the hepatitis C virus (HCV) life cycle have not been reproduced in cell culture, which has slowed research progress on this important human pathogen. Here, we describe a full-length HCV genome that replicates and produces virus particles that are infectious in cell culture (HCVcc). Replication of HCVcc was robust, producing nearly 10(5) infectious units per milliliter within 48 hours. Virus particles were filterable and neutralized with a monoclonal antibody against the viral glycoprotein E2. Viral entry was dependent on cellular expression of a putative HCV receptor, CD81. HCVcc replication was inhibited by interferon-alpha and by several HCV-specific antiviral compounds, suggesting that this in vitro system will aid in the search for improved antivirals.     

HLA and NK cell inhibitory receptor genes in resolving hepatitis C virus infection

Natural killer (NK) cells provide a central defense against viral infection by using inhibitory and activation receptors for major histocompatibility complex class I molecules as a means of controlling their activity. We show that genes encoding the inhibitory NK cell receptor KIR2DL3 and its human leukocyte antigen C group 1 (HLA-C1) ligand directly influence resolution of hepatitis C virus (HCV) infection. This effect was observed in Caucasians and African Americans with expected low infectious doses of HCV but not in those with high-dose exposure, in whom the innate immune response is likely overwhelmed. The data strongly suggest that inhibitory NK cell interactions are important in determining antiviral immunity and that diminished inhibitory responses confer protection against HCV.     

丙型肝炎病毒囊膜蛋白基因疫苗的构建及其诱导小鼠体液免疫应答

将丙型肝炎病毒（HCV）H77株全长囊膜蛋白基因克隆到真核表达载体pcDNA4.0的CMV启动子下游，采用磷酸钙转染法将重组质粒转入293T细胞，用流式细胞仪检测细胞瞬时表达的目的蛋白。将构建的重组质粒肌注BALB/c小鼠，用表达HCV囊膜蛋白的SP2/0细胞作为抗原，用流式细胞仪检测小鼠血清HCV囊膜蛋白抗体。再用原核系统表达的E2蛋白作为抗原,Western Blot检测免疫小鼠血清抗体。试验结果显示,囊膜蛋白E1E2在293T细胞膜上进行了瞬时表达，基因疫苗免疫小鼠后产生了抗HCV囊膜蛋白的抗体，该抗体能与表达HCV囊膜蛋白的SP2/0细胞特异性结合，Western Blot表明该抗体也能与原核系统表达的E2蛋白结合。     

HCV persistence and immune evasion in the absence of memory T cell help

Spontaneous resolution of hepatitis C virus (HCV) infection in humans usually affords long-term immunity to persistent viremia and associated liver diseases. Here, we report that memory CD4+ Tcells are essential for this protection. Antibody-mediated depletion of CD4+ Tcells before reinfection of two immune chimpanzees resulted in persistent, low-level viremia despite functional intra-hepatic memory CD8+ Tcell responses. Incomplete control of HCV replication by memory CD8+ Tcells in the absence of adequate CD4+ Tcell help was associated with emergence of viral escape mutations in class I major histocompatibility complex-restricted epitopes and failure to resolve HCV infection.     

河南省猪瘟流行毒的E2基因序列分析和基因分型

采用 RT- PCR和 n PCR扩增出 3株河南省近期 (1999～ 2 0 0 0年 )猪瘟流行野毒的 E2 基因 ,分别克隆于 PMD- 18T载体 ,对其进行了核苷酸序列测定及氨基酸序列推导 ,同时进行了同源性比较及 E2 糖蛋白结构的分析。结果表明 ,所测 3株 HCV E2 基因的长度均为 116 9bp。编码的氨基酸序列均包括完整信号肽序列和部分跨膜序列 ,共由 384个氨基酸组成。3株流行毒的 E2 基因核苷酸序列同源性在 99%以上 ,相应的氨基酸序列同源性也在 99%以上 ;这 3株流行毒与 C-株兔脾组织毒 (疫苗种毒 ) E2 基因的核苷酸序列同源性在 83.14 %～ 83.2 3% ,相应的氨基酸序列同源性在 88.14 %～ 88.6 8%。经遗传发生关系分析 ,C-株兔组织毒 C-株细胞毒属于组群 1(group 1) ,河南省近期流行猪瘟毒属于组群 2 (group 2 ) ,近期猪瘟流行毒与 C-株疫苗毒的 gp5 5蛋白之间存在一定的差异     

Regulation of interferon regulatory factor-3 by the hepatitis C virus serine protease

Persistent infections with hepatitis C virus (HCV) are likely to depend on viral inhibition of host defenses. We show that the HCV NS3/4A serine protease blocks the phosphorylation and effector action of interferon regulatory factor-3 (IRF-3), a key cellular antiviral signaling molecule. Disruption of NS3/4A protease function by mutation or a ketoamide peptidomimetic inhibitor relieved this blockade and restored IRF-3 phosphorylation after cellular challenge with an unrelated virus. Furthermore, dominant-negative or constitutively active IRF-3 mutants, respectively, enhanced or suppressed HCV RNA replication in hepatoma cells. Thus, the NS3/4A protease represents a dual therapeutic target, the inhibition of which may both block viral replication and restore IRF-3 control of HCV infection.     

Evolutionary Patterns of the Proviral gp90 V3 to V5 Regions of Equine Infectious Anemia Virus Associated with Immune Selection in Progressors and Nonprogressors

The aim of this study was to determine the genomic evolutionary pattern of virulent equine infectious anemia virus (EIAV)during persistent infection.The evolutionary dynamics of proviral genomes were examined by challenging an EIAV seronegative equine (pony 1) and three EIAV vaccinated equines (ponies 4,7,and 8) with the Chinese virulent strain EIAVL.Ponies I and 7 succumbed to disease and were called progressors,while ponies 4 and 8 lacked clinical symptoms and were considered nonprogressors.Sequences spanning the V3,V4,and V5 hyper-variable regions of the EIAV-L envelope gp90 gene were sequenced from each pony as evolutionary markers of the provirus.The proviral genome of the EIAV-Linoculum evolved during persistent infection and displayed different patterns between EIA progressors and nonprogressors.Inoculum-like variants were isolated from nonprogressors during persistent infection,but only from progressors during acute infection.Variant mutations from nonprogressors were dispersed throughout the sequenced region,while those from progressors were predominantly localized to V3.Humoral immunity and virus variant population selection analyses indicated that immune selection was positive in chronically infected progressors and weak in nonprogressors.In-frame stop codons were frequently localized to a defect "hot spot".The high number of defective variants in nonprogressors may promote disease survival.     

丙肝感染宿主亲嗜性基因OCLN和CD81在猕猴川西亚种组织中的分布

丙型肝炎动物模型缺乏限制了丙型肝炎的研究,如果与人类基因背景相似的猕猴能作为丙型肝炎动物模型将有重大意义。文章拟通过检测决定HCV感染物种特异性的关键基因OCLN与CD81在猕猴不同组织的表达量,并以HCV易感的人肝癌细胞系Huh 751为对照,来探究猕猴作为丙型肝炎动物模型的可能性。结果表明,OCLN与CD81在猕猴川西亚种的肝脏、脾脏、肺脏、淋巴结和脊髓中均有表达,但表达量均极显著低于在Huh 751细胞系中的表达量（P<0.01）。肝脏中的OCLN表达量略低于肺脏中的表达量,而肝脏CD81的表达量均高于其他组织。并且,OCLN在不同组织中的表达水平与人体OCLN表达规律相符合（脾＜肝＜肺）。该结果提示HCV感染的宿主亲嗜性基因OCLN与CD81在猕猴中的低表达量可能导致HCV不能有效地结合组织靶细胞,因此推测,猕猴不能直接用于丙型肝炎动物造模可能与此有关。今后可通过转基因等技术增强OCLN与CD81基因的表达,从而使猕猴获得直接感染HCV的能力。     

流式细胞术检测TcR Vα24/Vβ11双阳性NKT细胞及其在HCV感染者中的应用

目的研究丙肝病毒感染者外周血NKT细胞含量与HCV感染状态之间的相关性.方法应用流式细胞术检测TcR Vα24/Vβ11双阳性NKT细胞.结果抗-HCV阳性且HCVRNA也阳性的病例NKT细胞数量明显低于抗-HCV阳性而HCV RNA阴性的病例（P〈0.01）,明显低于正常对照组（P〈0.05）.结论由于NKT细胞数量减少或活化受阻,造成HCV的持续感染,不利于病毒的清除;另一方面,由于肝内有HCV的存在,NKT细胞大量向肝内趋化,导致外周血的数量相对减少.     

广西猪瘟流行毒与C-株疫苗毒gp55（E2）主要抗原区DNA序列差异的比较

采用反转录 PCR(RT- PCR)和套式 PCR(nest Polym erase Chain Reaction,n PCR)扩增出 3株广西省近期 (1999～ 2 0 0 0年 )猪瘟流行野毒的 E2 基因 ,将其分别克隆于 PMD- 18T载体上并进行了核苷酸序列测定 ,根据 C株及 Brescia和 Alfort株确定起始氨基酸三联体的正确位置后进行氨基酸序列推导 ,同时进行了同源性比较及 E2 糖蛋白结构的分析。结果表明 ,所测 3株 HCV E2 基因的长度均为 1170 bp,编码的氨基酸序列均包括完整信号肽序列和部分跨膜序列 ,共由 384个氨基酸组成。3株流行毒的 E2 基因核苷酸序列同源性为 90 .10 %～ 98.5 4%,相应的氨基酸序列同源性为 89.5 9%～ 97.92 %。这 3株流行毒与 C-株兔脾组织毒 (疫苗种毒 ) E2 基因的核苷酸序列同源性为 82 .87%～ 83.99%,相应的氨基酸序列同源性为 86 .98%～ 90 .10 %,表明近期猪瘟流行毒与 C-株疫苗毒的 gp5 5蛋白之间存在一定的差异。     

广西猪戊型肝炎血清流行病学调查

为了解广西各地猪群戊型肝炎（HE）的感染状况及其流行特点,采用双抗原夹心ELISA检测来自广西11个市34个猪场及南宁市某屠宰场猪血清中的抗HEV抗体,并比较分析不同地区、不同年龄段猪抗HEV抗体阳性率的差异。结果表明,所检测的34个猪场均为抗HEV抗体阳性猪场;537份血清中有448份呈阳性,阳性率为83．4％;其中2月龄以内、3-4月龄和5月龄以上猪的抗体阳性率分别为74．0％、47．0％和95．5％,差异显著（P〈0．05）;广西各地区猪群抗HEV抗体阳性率均在70．0％以上,差异不显著（P〉0．05）。可见,广西猪群普遍存在HEV感染,且母源抗体消失后,猪群抗HEV抗体阳性率随着年龄增长而升高。     

猪瘟伪狂犬病重组病毒SA215(A)疫苗株的构建(初报)

采用磷酸钙转染系统 ,将伪狂犬病三基因缺失疫苗SA2 15株DNA与PP6 3LacZE2DNA共转染Vero细胞 ,获得SA2 15 (A) 1、SA2 15 (A) 2和SA2 15 (A) 3等 12个重组病毒株。以光生物素标记的HCVE2基因为探针进行初步鉴定后 ,挑选SA2 15 (A) 1株作BamHⅠ酶切和southern转印杂交鉴定 ,结果表明构建是成功的 ,将其命名为SA2 15(A)。直接荧光抗体检测、SDS -PAGE电泳和western免疫印迹检测结果表明HCVE2基因在重组病毒内获得表达 ,产生大小约 5 1kd的蛋白。对SA2 15 (A)株进行部分生物学特性研究 ,培养特性观察试验表明该毒株可适应Vero、BHK2 1和鸡胚成纤维细胞等多种细胞 ,但对不同细胞系表现有一定的差异。     

肝病患者纤维蛋白溶解系统关键因子检测的意义

目的研究肝病患者血浆中继发性纤维蛋白溶解系统中重要标志物D-二聚体（D-D）和纤维蛋白原（FIB）指标的变化及其临床意义.方法采用美国ACL-ELITEPRO全自动凝血仪检测35例健康体检者和94例不同肝病患者血浆中FIB和D-D含量.结果与健康体检正常组相比,各肝病组FIB含量降低,顺序为急性肝炎组〉健康对照组〉慢性肝炎组〉肝癌组〉肝硬化组.急性肝炎组和慢性肝炎组差别不显著,肝硬化组和肝癌组差别显著,差异有统计学意义（P〈0.05）;D-D含量升高顺序为健康对照组〈急性肝炎组〈慢性肝炎组〈肝癌组〈肝硬化组,各肝病组差异均有统计学意义.肝癌组和肝硬化组分别与急性肝炎组、慢性肝炎组和肝硬化组FIB和D-D含量比较均有显著差异,差异有统计学意义.结论血浆FIB和D-D可客观地反映肝病的严重程度.这为临床肝脏疾病的诊断、治疗以及判断预后提供了重要依据.     

丙型肝炎感染者IL-2、IL-6、IL-18血清水平的研究

目的:探讨血清IL-2、IL-6、IL-18水平与丙型肝炎病变的关系。方法:运用EL ISA法检测不同类型丙型肝炎感染者血清3种细胞因子IL-2、IL-6、IL-18的水平,观察感染者细胞因子水平与肝炎发展的关系。结果:慢性及重症肝炎感染者血清IL-2水平出现显著性下调(P<0.05),而慢性及重症肝炎感染者的IL-6、IL-18的水平均显著高于正常对照组(P<0.05或P<0.01),升高程度与病情相关。结论:血中细胞因子水平是判断丙肝感染者免疫状态及进行治疗的重要指标。     

Rat resistance to schistosomiasis: platelet-mediated cytotoxicity induced by C-reactive protein

In rats infected with the parasite Schistosoma mansoni, the concentration of C-reactive protein in the serum increases after the lung stage of infection and is at its highest at the time of terminal worm rejection. The peak of platelet-mediated cytotoxicity induced by infected serum that has been heated (and is free of immunoglobulin E) as well as the time course for the development of platelet cytotoxic activity in infected rats was found to be correlated with the concentration of C-reactive protein. Rat and human platelets treated with homologous serum obtained during an acute phase of inflammation or with purified C-reactive protein were able to kill the immature forms of the worm in vitro. Platelets treated with C-reactive protein were furthermore capable of conferring significant protection against schistosomiasis in transfer experiments. Collectively these data indicate that a system that includes C-reactive protein and platelets participates in the natural resistance of the rat to schistosomal infection.     

乙型肝炎与戊型肝炎病毒重叠感染者血清HBV标记物结果分析

目的：观察乙型肝炎病毒（ＨＢＶ）与戊型肝炎病毒（ＨＥＶ）重叠感染对ＨＢＶ复制的影响。方法：以血清ＨＢｓＡｇ和抗ＨＥＶ均阳性患者３６例作为重叠组,以单纯ＨＢＶ感染患者３６例作为对照组。两组病例抗ＨＡＶ－ＩｇＭ和抗ＨＣＶ均为阴性。采用ＥＬＩＳＡ法测定两组的ＨＢＶ－Ｍ。结果：重叠组血清ＨＢｅＡｇ的阳性率明显低于对照组（Ｐ＜０．００１）,抗ＨＢｅ阳性率明显高于对照组（Ｐ＜０．０５）。结论：ＨＢＶ与ＨＥＶ重叠感染后,ＨＢＶ的复制受到一定程度的抑制     

丙型肝炎病毒囊膜蛋白基因E1E2在真核细胞中的表达及动物免疫试验

利用DNA重组技术将丙型肝炎病毒（HCV）H77株E1E2囊膜蛋白基因插入逆转录病毒载体pBABE-puro中，构建成重组逆转录病毒载体pBABE-puro-E1E2，用重组逆转录病毒载体与pVSVg质粒磷酸钙共沉淀法转染293T细胞,包装逆转录病毒假病毒。用包装的假病毒感染SP2/0细胞，经嘌呤霉素筛选阳性细胞后进行流式细胞仪技术（FACS）分析，结果表明,HCV E1E2基因在SP2/0细胞膜上成功表达。将表达E1E2蛋白的SP2/0细胞腹腔免疫BALB/c小鼠，经FACS分析免疫鼠血清，成功诱导小鼠产生了抗HCV E1E2蛋白的抗体，Western blot检测结果表明该抗体能与原核系统表达的E2蛋白结合。     

病毒性肝炎患者血清微量元素的检测及其临床意义

目的:探讨病毒性肝炎患者和肝炎肝硬化患者血清微量元素水平与病情的相关性。方法:采用RI- 10 0 0 TM全自动生化分析仪检测2 73例病毒性肝炎患者(慢性肝炎130例、急性肝炎35例、重型肝炎38例、肝炎肝硬化70例)及4 0例正常对照者血清中镁、铁、铜、锌的水平。结果:各型肝炎及肝硬化患者血清锌水平均显著低于正常对照组(P<0 .0 1)。除急性肝炎组外,慢性肝炎、重型肝炎、肝炎肝硬化患者血清中镁、铜水平均显著低于正常对照组(P<0 .0 1) ,血清中镁、铜、锌水平以重型肝炎组最低。各型肝炎及肝硬化患者血清铁水平均显著高于正常对照(P<0 .0 1) ,以重型肝炎组最高。结论:检测病毒性肝炎患者血清中的微量元素,对了解病情及指导临床治疗具有一定的意义。