Hepatitis viruses and hepatocarcinogenesis

Hepatocellular carcinoma (HCC) is among the most frequent malignancies worldwide. Hepatitis viruses, such as the hepatitis B virus (HBV) and hepatitis C virus (HCV) are undoubtedly listed in the etiology of HCC. Studies show that, in the near future, viral hepatitis will carry increasing weight in the etiology of HCC. This review briefly discusses the known carcinogenic effects of HBV and HCV in the light of experimental and human studies. The data show that viral proteins may directly interfere with gene products responsible for cell proliferation and cell growth. Many other signal transduction cascades may be affected as well. Direct integration of HBV viral sequences into the host genome increases the genomic instability. The genomic imbalance allows the development and survival of malignant clones bearing defected genomic information. HBV and HCV infection induces indirect and direct mechanisms through cellular damage, increased regeneration and cell proliferation, therefore enhancing the development of HCC.     

Seroepidemiology of hepatitis C virus infection in Japan and HCV infection in haemodialysis patients

Abstract: Since January 1990, Japanese Red Cross Blood Centres have introduced hepatitis C virus screening with a first-generation ELISA. From April to December 1992, approximately 0.98% among 10905 489 blood donations screened by a second-generation assay were anti-HCV-positive in all Japan. Seropositivity of anti-HCV increased with the age and serum transminase value in both sexes. In blood donors having a history of transfusion, the anti-HCV reactive rate was 7.4%. The results of the study made by the Japanese Red Cross Non-A, Non-B Hepatitis Research Group show the effectiveness of implementation of HCV screening to prevent posttransfusion hepatitis. Consecutive haemodialysis patients with chronic renal failure are at risk for inflection by a variety of blood-borne agents transmitted within dialysis units. Because of their immunocompromised state, they frequently also have an unusual susceptibility to a variety of nosocomial infections, such as HBV, and HTLV-I. We tested the prevalence of anti-HCV in 1423 (848 males and 575 females) haemodialiysis patients from 18 hospitals in Kumamoto Prefecture, Japan using the Orhto first generation anti- HCV screening assay. There were 316 patients (22.2%) positive for HCV antibodies. The second-generation test was positive in most haemodialysis patients who were eractive to the firs-generation assay. The prevalence of HCV infection increased with the duration of haemodialysis, yet there was a high frequency of HCV seropositivity even wihtout blood transfusion. Acquisition of HCV in dialysis patients could be explained by HCV seropositivity even without blood (all haemodialysis are done with disposable kits, and needles), by secondary HCV infection after the immunodeficiency of haemodialysis, or by HCV infection of the kidney or glomerular deposition of immune HCV/anti-HCV complexes leading to chronic renal failure (as with HBV infection of the liver and kidney).     

常见肝炎病毒感染性疾病相关miRNA功能研究进展

病毒感染引发的疾病一直威胁着人类健康。Mi RNA是真核生物表达的一类重要的小分子RNA,可特异性的调节基因与蛋白的表达。mi RNA的研究为病毒性疾病的发生发展提供了新思路,为目前热点研究领域。随着mi RNA的研究深入,一些病毒感染中相关mi RNA的功能也被相继报道,如有些mi RNA具有抑制病毒感染宿主细胞的功能,有些mi RNA则可促进病毒在宿主细胞中的复制,有些mi RNA却参与病毒相关疾病的发生,还有些mi RNA则可作为病毒感染性疾病的特异性生物标志物。本文主要以两种常见肝炎病毒:HBV、HCV为例来系统阐述mi RNA在病毒感染中的相关功能。     

一种新型丙型肝炎病毒培养方法的建立

以丙肝病毒 (ＨＣＶ)为切入点 ,建立一套通过基因操作技术在体外细胞培养系统中生产高滴度反转录病毒颗粒的大规模病毒培养技术平台 ,并将该技术应用于其它难以体外大规模培养的反转录病毒的生产。该体系包括一株插入Ｔ7ＲＮＡ聚合酶基因的重组痘苗病毒ｖＴＦ 3和 2个重组质粒 ,质粒ＰＴ7ＨＣＶ在上游Ｔ7启动子和Ｔ7终止子之间插入ＨＣＶ基因组ｃＤＮＡ ,可通过Ｔ7ＲＮＡ聚合酶指导转录产生 9 5ｋｂ的ＨＣＶＲＮＡ ;另一质粒Ｐ...     

Polyclonal T-cell activation protocol stimulates preferential expansion of EBV-specific T-cell clones in vitro

Purpose Allogeneic bone marrow transplantation (AlloBMT) can be curative for patients with leukemia. The most important anti-leukemic effect may be mediated by the T-cells contained within the graft; however, the allogeneic T-cells may also give rise to graft-vs-host disease (GVHD). One way to control GVHD might be to transduce the donor T-cells with a drug-inducible suicide gene. If a retrovirus vector is to be used for this transduction, activation of the T-cells is required for integration of the transgene to occur. The activation protocol should ensure expansion of a broad repertoire of donor T-cells. Notably, T-cells specific for herpes virus family antigens are important for adoptive immunoprotection.Methods To define optimal activation conditions for retrovirus-mediated suicide gene transduction of donor T-cells, we examined the repertoire of CD8+ T-cells in general, and Epstein-Barr virus (EBV) specific T-cells in particular, following two different activation and expansion procedures.Results We found that repeated CD3/CD28 stimulation resulted in a high level of activation-induced T-cell death, affecting in vivo expanded clones, some of which were specific for EBV, in particular. In contrast, initial CD3/CD28 activation followed by proliferation in interleukin-2 lead to expansion of EBV-specific clones over and above the expansion observed for CD8+ T-cells in general.Conclusion These results should impact on protocols for ex vivo activation of T-cells prior to suicide gene transduction.     

C3/Ig and Ig/C3 two-component-determined circulating immune complexes (TCIC) in patients with HCV infection

In the present study, we measured the levels of immunoglobulin (Ig)- and complement 3 (C3)-determined circulating immune complexes (two-component-determined CIC, or TCIC) in hepatitis C virus (HCV)-infected patients. TCIC was dissected into C3/Ig-TCIC and Ig/C3-TCIC by a reciprocal use of coating and detecting antibodies. The current study was carried out in 117 infected HCV patients and 252 healthy controls. We found that C3/Ig-TCIC elevation was a common feature in patients with HCV infection. Positive rates and levels of C3/IgG-TCIC and C3/IgM-TCIC were significantly higher in the patients with abnormal alanine aminotransferase (ALT) than patients with normal ALT (70.6% vs. 17.0%, 0.56 OD vs. 0.47 OD and 0.71 OD vs. 0.65 OD, respectively, P<0.001). However, the levels of IgM/C3-TCIC and IgA/C3-TCIC were significantly higher in individuals with HCV infection than in healthy controls, whereas the level of IgG/C3-TCIC was significantly lower in the former group than in the latter group. In summary, our results suggest that IgG and C3 TCIC may play an important role in liver cell injury during the course of HCV infection and may be a hallmark for hepatitis C pathogenesis. Elevated C3/Ig-TCIC, accompanied by decreased Ig/C3-TCIC, forms a peculiar trait in HCV infection. Our findings thus provide new insights into HCV pathogenesis.     

ELISA in serodiagnosis of HCV infection

Abstract: A high level of anti-HCV is generally associated with viral replication and the number of recognized epitopes appears to be correlated with the viral charge. Nevertheless, the absence of detectable antibodies in about 60% of patients during the acute phase of the disease adn in 10% of chronically infected (generally immunocompromised subjects) are heavy handicaps for HCV serology. Moreover, low levels of anti-HCV antibodies can persist after complete recovery, and HCV viremia does not appear to be associated with the presence of a special antibody specificity. The immunoblots presented as 'confirmatory test' always appear to be less sensitive that the screening tests adn therefore are unable to discriminate between post-infection antibodies and false-positive reactions, as rare as they can be. In these cases, as in non-responder patients, PCR appears essential. The possible reasons of immune response limitations and the possible improvements of HCV serology are discussed.     

乙肝病毒感染与B 细胞型非霍奇金淋巴瘤的相关性研究

目的：探讨乙肝病毒（HepatitisB virus, HBV）感染与B细胞型非霍奇金淋巴瘤（B-cell Non-Hodgkin''s Lymphoma,B-cell NHL）的相关性。方法：回顾性分析2008 年1 月至2014 年1 月我院232 例NHL患者作为研究组,另选取经病理学、影像学等诊断为其他类型肿瘤的患者230 例作为对照组。比较两组研究对象HAV、HBV及HCV 的感染情况。结果：研究组患者HBsAg阳性率高于对照组,差异具有统计学意义（P<0.05）;但两组患者Anti-HAV和Anti-HCV 阳性率无明显差异（P>0.05）。B-cell NHL患者HBsAg阳性率高于对照组和T-cell NHL患者,差异具有统计学意义（P<0.05）。T-cell NHL患者HBsAg阳性率与对照组无显著差异（P>0.05）。B-cell NHL 年轻患者HBsAg 阳性率高于T-cell NHL患者,差异具有统计学意义（P<0.05）。B-cell NHL 患者Anti-HBs、HBeAg、Anti-HBe及Anti-HBc阳性率与对照组存在明显差异（P<0.05）;而T-cellNHL患者与对照组无显著差异（P>0.05）。结论：B-cellNHL感染HBV 的几率较高,HBV 感染与B-cell NHL早期发病有明显的关联性。     

丙型肝炎病毒多表位DNA疫苗的研究进展

丙型肝炎病毒（hepatitis C virus,HCV）是非甲非乙型肝炎的主要病原,目前还没有有效的预防和治疗手段。多表位DNA疫苗(multi-epitope DNA vaccine, minigenes/epigenes)是指通过筛选与组合优势抗原表位(包括T细胞、B细胞表位)基因,以能产生高效细胞、体液免疫应答进而清除HCV病毒为目的的新型核酸疫苗。其优势在于通过选择最具免疫保护潜力的表位以覆盖尽量多的病毒亚型和诱导全面的机体抗病毒免疫应答,并尽量减少无关、干扰和抑制序列可能产生的负面影响。本文介绍近年来国内外在丙型肝炎多表位DNA疫苗方面的研究进展,并展望了其发展方向。     

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.     

丙型肝炎病毒高水平复制细胞株的构建及其机制初步研究

旨在探讨丙型肝炎病毒(hepatitis C virus, HCV)cured细胞株的易感机制。本研究将体外转录的HCV RNA电转入肝癌细胞系Huh 7细胞,建立HCV复制子细胞株,用 γ-干扰素(interferon,IFN)处理复制子细胞株,获得HCV cured Huh 7A和Huh 7B细胞株。用插入报告基因的HCV毒株Jc1-G感染上述细胞株,分别进行荧光素酶活性测定、蛋白质印迹法和荧光定量聚合酶链反应(polymerase chain reaction,PCR)检测以验证其易感性。收集Huh 7、Huh 7.5、Huh 7A和Huh 7B细胞并利用IFN-α处理,之后用蛋白质印迹法及荧光定量PCR进行检测,验证细胞株中IFN诱生信号通路中关键因子内源性表达及抗病毒活性ISGs的激活水平。结果显示,在Huh 7A和Huh 7B细胞中检测不到病毒RNA,与Huh 7细胞一致。病毒感染实验中,与Huh 7细胞相比,Huh 7A和Huh 7B细胞株中荧光素酶活性增高百倍,病毒蛋白表达和RNA水平亦显著上调,与Huh 7.5细胞株中的表达水平接近。IFN信号通路实验中,与Huh 7细胞相比,Huh 7A和Huh 7B细胞株中RIG-I/MDA5/MAVS内源性蛋白表达和mRNA水平无明显差异;IFN-α处理细胞后IFN刺激基因isg56,mx1,mx2,oax1,oax2,viperin,cxcl10,ifitm1和ifitm3激活水平亦无显著变化。结果提示,本研究制备的Huh 7A和Huh 7B细胞株可支持HCV高水平复制,将为研究病毒复制机制提供有力的支持。     

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…     

一株丙型肝炎病毒缺陷株NS5A基因片段的序列分析

Ｑ丙型肝炎是由丙型肝炎病毒（ＨＣＶ）引起的一种严重的传染病。丙型肝炎病毒主要通过输血和应用不洁的血液制品传播,所以利用敏感、特异的检测方法筛选献血员对丙型肝炎的预防尤为重要。现在第二代ＨＣＶ检测试剂已大批应用,加入ＮＳ５Ａ抗原的第三代试剂也正在研制中...     

戊肝与丙肝病毒在献血员人群中感染状况的对比研究

采用市售试剂对武汉地区乡村献血员进行血清抗ＨＥＶ与抗ＨＣＶ检测,两者的阳性率分别为５．７４％及９．３５％。在２８８份有ＡＬＴ记录的单采浆献血员中,有近期ＡＬＴ升高史的献浆者抗ＨＥＶ及抗ＨＣＶ检出率分别为１４．０４％及１４．１８％,均显著高于无近期ＡＬＴ升高史的献浆者。对上述标本同时进行多项血清ＨＢＶ标志检测,抗ＨＥＶ阳性及抗ＨＣＶ阳性组献血员多项ＨＢＶ标志检测结果与相应阴性组比较均未见显著的差别。     

4′-Substituted pyrimidine nucleosides lacking 5′-hydroxyl function as potential anti-HCV agents

Hepatitis C virus (HCV) infection is one of the major health problems worldwide. If left untreated, it leads to liver cirrhosis, liver cancer and death. Herein, we report synthesis and anti-HCV activity of a new class of pyrimidine nucleosides possessing a 4′-carboxymethyl (9–16, 21 and 23) or 4′-carboxamide function (17–19 and 24). Among these, 10–12 (EC₅₀ = 33.1–42.4 μM), 14 and 21 (EC₅₀ = 43.4–59.5 μM) exhibited potent activity in HCV-1a replicon cells without any toxicity to parent Huh-7 cells (CC₅₀ = >829–1055 μM). The anti-HCV activities demonstrated by this unusual class of compounds were superior to that of ribavirin (EC₅₀ = 81.9 μM). Further, the most active analog, 12, was found to interact synergistically with ribavirin to inhibit HCV RNA replication.