乙型肝炎病毒感染的自然病程

在我国,慢性乙型肝炎病毒表面抗原（HBsAg）携带者达1.2亿人之多,其中相当一部分患者有活跃的病毒复制和肝脏炎症反应,而且部分患者会演进为肝硬化和/或肝细胞癌,这对我国人民身体健康和国民经济危害极大.     

乙型肝炎病毒在异种动物原代肝细胞中复制与表达的研究

目的 探讨乙型肝炎病毒(HBV)DNA复制和表达的跨种属特异性。 方法 分离培养原代大鼠肝细胞(PRH)与原代鸭肝细胞(PDH),电转HBV线性裸DNA(转染组每1×107PRH或PDH 1.19×1012拷贝),分别于转染后1～15d各时点,收集PRH或PDH培养上清液与细胞裂解液,分别以Southern杂交分析和斑点杂交法分析HBV DNA的复制中间体与复制型式;以全自动免疫荧光检测系统检测乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg),以western免疫印迹、免疫斑点印迹和免疫细胞化学法检测乙型肝炎核心抗原(HBcAg);用逆转录聚合酶链反应检测HBV S/X mRNA。以单纯电击PRH/PDH为对照组。 结果 Southern杂交分析显示:PRH/PDH转染组HBV DNA均为游离复制型,可见4.0 kb以下分子区带,包括松弛环状DNA、共价闭环形DNA和单链线形DNA等复制中间体;未见整合型HBVDNA-高分子(4.0～24.0 kb)区带。HBV DNA在PRH中表达蛋白产物水平,HBsAg于转染后各时点PRH裂解液中均可检测到(P/N值2.17～93.41,平均值为14.74±31.82,阳性≥2.1),峰值于1～3 d;仅转染后1d PRH培养上清液中检测到HBsAg,P/N值为6.66;HBcAg和HBeAg仅于转染后1～3 d时点内检测到低度表达;HBV S mRNA为阳性,而X mRNA为阴性。HBV DNA转染PDH组,转染后1、3、5 d各时点PDH裂解液中HBsAg分别为15.24、     

乙型肝炎病毒Ⅱ型与肝细胞癌

调查了237例肝细胞癌(HCC)患者血清中感染乙型肝炎病毒(HBV)的状态,其中152例(64.14%)HBs Ag阳性,并揭示HCC患者血清中存在HBV_2感染的血清学模式,即HBsAg阳性而抗-HBc阴性,占HBsAg阳性HCC患者的10.53%(16/152)。占所有HCC患者的6.75%(16/237)。应重视对HBV_2感染的防治。     

乙型肝炎病毒在HepG2细胞中的稳定复制及表达

目的 建立HBVHepG2肝细胞株，使HBV能长期稳定地表达抗原并复制。方法将含完整转录单位的1．2倍体HBVDNA经Sal1位点克隆入真核表达载体pREP10，构建的重组载体pREP—HBV以Lipofemamine2000转染HepG2细胞，250μg／ml。潮霉素筛选抗性细胞克隆。ELISA检测细胞上清液中的HBsAg和HBeAg。电子显微镜下观察细胞上清液HBV颗粒。制备HBV特异性探针，以Southern印迹法检测细胞株内HBV核心颗粒DNA。结果获得含1．2倍体HBVDNA的重组载体，即pREP—HBV，该重组载体转染HepG2细胞后，获得5株潮霉素抗性细胞RHBV1～RHBV5。均能表达HBsAg和HBeAg。Southern印迹结果显示各株细胞均可见明显的杂交拖带，即存在HBVDNA复制中间体。细胞培养上清液浓缩后在电子显微镜下可见成簇的、直径约42nm的HBV颗粒及直径为22～26nm的球形颗粒。结论 成功建立了HBV稳定复制及表达的HepG2细胞株，目前细胞已传代50次，每3天1次。     

原发性肝细胞癌乙型肝炎病毒X抗原表达的分析

我们前已报道了抗ＨＢＶＸ抗原（ＨＢｘＡｇ）三种不同肽段的抗体检出肝癌细胞中ＨＢｘＡｇ的存在，我们用ＰＣＲ技术扩增肝癌组织中，ｘ基因，用重组ＨＢｘＡｇ及其抗体中的抗ＨＢｘ和ＨＢｘＡｇ并用免疫组织化学的方法分析肝组织中ＨＢｘＡｇ的表达及其分布，我们发现ＨＢｘＡｇ在肝癌肝组织中阳性率高达９２．６％，而且分布广泛，主要表达在细胞浆，但细胞核内亦有见到，癌旁组织检出率也很高，而在小胆管上皮细胞内亦见有ＨＢｘ     

乙型肝炎病毒感染的肝癌患者肝组织中乙型肝炎病毒cccDNA的检测

HBV属嗜肝DNA病毒家族,其基因组是双链松弛环状DNA(rcDNA),相对分子量为(1.6～2.0)×109,全长约为3.2 kb。在HBV的生命周期中,共价闭合环状DNA (cccDNA)是HBV的原始复制模板,对HBV的复制及感染状态的建立具有十分重要的意义。清除HBV cccDNA是目前抗HBV药物研究的一个重要目标。我们对天津市第三中心医院收治的20例HBV感染的肝癌患者肝组织(包括癌     

乙型肝炎病毒和丙型肝炎病毒对纤维连接蛋白基因表达的调节

乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)感染是病毒性肝炎的主要病原体，感染后引起慢性化的频率较高。在我国，HBV或HCV的感染也是肝硬化和肝细胞癌(HCC)发病的主要原因。纤维连接蛋白(Fibronectin，Fn)是一种存在于血液、体液及各种组织中的具有多种功能的糖蛋白，来源于肝细胞，枯否细胞和内皮细胞，通过与整合素结合调节细胞间的粘附、免疫、凝血和血小板的聚集^[1]，另外在肿瘤与纤维化的发展与发病机制中，Fn也起着重要的作用^[2]。     

乙型肝炎病毒X蛋白在肝细胞癌发生中的作用机制

HBx蛋白（HBxAg）被认为是乙型肝炎病毒（HBV）致肝细胞癌（HCC）的病毒蛋白,能影响肝细胞的生长、转化、迁移和凋亡以及DNA的修复等过程,己成为近年来研究的热点之一。HBx是一种多功能的病毒蛋白,有转录调控作用,对肝细胞内许多癌基因和／或抑癌基因的表达有直接或间接的影响,在肝癌形成中起重要作用。通过对HBx在肝癌发生、发展中机制的深入研究,有望在HCC防治方面开辟新途径。     

中老年肝细胞癌患者乙型肝炎病毒X基因检测与分析

目的检测慢性乙型肝炎病毒(HBV)感染后肝细胞癌(HCC)患者血清HBVX基因,并与慢性乙型肝炎、肝硬化患者HBVX基因检出率对比分析。方法在微孔板上包被HBVX基因片段的捕获探针;PCR扩增被检标本中目的片段,其引物用Bio-Ⅱ-dUTP修饰,在微孔板上杂交后用链霉亲和素碱性磷酸酶系统显色。结果该方法检测HBVX基因灵敏,特异;乙型肝炎后HCC患者HBVX基因检出率明显高于慢性乙型肝炎组和肝硬化组。结论检测HBVX基因对HCC具有辅助诊断价值。     

过氧化物酶体增殖物活化受体与乙型肝炎病毒的复制及肝纤维化的关系

过氧化物酶体增殖物激活受体(peroxisome proliferator activated receptor，PPAR)是一种由配体激活的核转录因子。近年研究发现。PPAR能够调节炎症反应、控制细胞周期和细胞凋亡，其中也包括一些肝炎相关疾病，如乙型肝炎病毒(HBV)的复制、肝纤维化以及肝癌等。本文主要论述。PPAR与HBV复制、肝纤维化的关系。     

Occult hepatitis B virus and hepatocellular carcinoma

Occult hepatitis B virus (HBV) infection (OBI) is a challenging pathobiological and clinical issue that has been widely debated for several decades. By definition, OBI is characterized by the persistence of HBV DNA in the liver tissue (and in some cases also in the serum) in the absence of circulating HBV surface antigen (HBsAg). Many epidemiological and molecular studies have indicated that OBI is an important risk factor for hepatocellular carcinoma (HCC) development. OBI may exert direct pro-oncogenic effects through the activation of the same oncogenic mechanisms that are activated in the course of an HBsAg-positive infection. Indeed, in OBI as in HBV-positive infection, HBV DNA can persist in the hepatocytes both integrated into the host genome as well as free episome, and may maintain the capacity to produce proteins-mainly X protein and truncated preS-S protein - provided with potential transforming properties. Furthermore, OBI may indirectly favor HCC development. It has been shown that the persistence of very low viral replicative activity during OBI may induce mild liver necro-inflammation continuing for life, and substantial clinical evidence indicates that OBI can accelerate the progression of liver disease towards cirrhosis that is considered the most important risk factor for HCC development.     

Key role of hepatitis B virus mutation in chronic hepatitis B development to hepatocellular carcinoma

Chronic hepatitis B virus (HBV) infection is a major risk factor for hepatocellular carcinoma (HCC). The HBV mutations, which include point mutation, deletion, insertion and truncation mutation of HBV gene in 4 open reading frames (S, C, P, X), are closely associated with HCC pathogenesis. Some mutations accumulated during chronic HBV infection could be regarded as a biomarker to predict the occurrence of HCC. The detection of the mutations in clinical practice could be helpful for defining better preventive and therapeutic strategies and, moreover, predicting the progression of liver disease.     

Natural history of chronic hepatitis B:Phases in a complex relationship

Chronic hepatitis B(CHB)is a condition of globalprevalence and its sequelae include cirrhosis and hepatocellular carcinoma.The natural history of CHB isa complex interplay of virological,environmental andhost factors.The dynamic relationship between thevirus and host evolves over the duration of the infection and different phases of the disease have been observed and described.These have been conceptualizedin terms of the state of balance between the host immune system and the hepatitis B virus and have beengiven the labels immune tolerant,immune clearance,immune control and immune escape although othernomenclature is also used.Host factors,such as age atinfection,determine progression to chronicity.Virological factors including hepatitis B viral load,mutationsand genotype also have an impact on the adverseoutcomes of the infection,as do hepatotoxic cofactorssuch as alcohol.Our understanding of the natural history of CHB has evolved significantly over the past fewdecades and characterizing the phase of disease ofCHB remains an integral part of managing this virus in the clinic.     

慢性乙型肝炎病毒感染者肝细胞癌筛查和监测

肝细胞癌是我国常见恶性肿瘤,疾病负担十分沉重。筛查和监测是提高肝细胞癌患者早诊早治和生存率的有效措施。慢性乙型肝炎病毒感染是我国肝细胞癌的主要病因,有必要制定专门的筛查和监测策略。中国肝炎防治基金会组织国内有关专家,参考国内外相关指南,并结合当前研究进展和临床实践经验共同讨论后达成一致意见,旨在为规范开展慢性乙型肝炎病毒感染者肝细胞癌的筛查和监测提供参考,进而改善我国肝细胞癌的防控效果和患者预后。     

Expression of B7-H4 and hepatitis B virus X in hepatitis B virus-related hepatocellular carcinoma

AIM: To investigate the expression and clinical significance of B7-H4 and hepatitis B virus X(HBx) protein in hepatitis B virus-related hepatocellular carcinoma(HBV-HCC).METHODS: The expression of B7-H4 in the human HCC cell lines Hep G2 and Hep G2.2.15 were detected by western blot, flow cytometry, and immunofluorescence. The expression of B7-H4 and HBx in 83 HBV-HCC was detected by immunohistochemistry, and the relationship with clinicopathological features was analyzed. Paraffin sections were generated from 83 HBV-HCC patients(22 females and 61 males) enrolled in this study. The age of these patients ranged from 35 to 77 years, with an average of 52.5 ± 11.3 years. All experiments were approved by the Ethics Committees of the Second Affiliated Hospital, Zhejiang University School of Medicine.RESULTS: B7-H4 was significantly upregulated in Hep G2.2.15 cells compared to Hep G2 cells. Specifically, the protein expression of B7-H4 in the lysates of Hep G2 cells was more than that in Hep G2.2.15 cells. In addition, HBx was expressed only in Hep G2.2.15 cells. Similar data were obtained by flow cytometry. The positive rates of B7-H4 and HBx in the tissues of 83 HBV-HCC patients were 68.67%(57/83) and 59.04%(49/83), respectively. The expression of HBx was correlated with tumor node metastases(TNM) stage, and the expression of B7-H4 was positively correlated with HBx(rs = 0.388; p 0.01). The expression level of B7-H4 in HBx-positive HBV-HCC tissues was substantially higher than that in HBx-negative HBV-HCC tissues. The expression level of B7H4 was negatively related to tumor TNM stage.CONCLUSION: Higher expression of HBx and B7-H4 was correlated with tumor progression of HBV-HCC, suggesting that B7-H4 may be involved in facilitating HBV-related hepatocarcinogenesis.     

Human hepatitis B virus and hepatocellular carcinoma II. Experimental induction of hepatocellular carcinoma in tree shrews exposed to hepatitis B virus and aflatoxin B1

On the basis of the successful establishment of an animal model in tree shrews experimentally infected with human hepatitis B virus (HBV), a study on the hepatocarcinogenic effects of HBV and/or aflatoxin B1 (AFB1) was conducted. The results showed that the incidence of hepatocellular carcinoma (HCC) was significantly higher in the animals both infected with HBV and exposed to AFB1 (52.94%) than in those solely infected with HBV (11.11%) or exposed to AFB1 (12.50%). No HCC of precancerous lesions were found in the controls that were neither HBV-infected nor AFB1-exposed. Precancerous lesions, including liver cell dysplasia and enzyme-altered hyperplastic hepatocyte foci, were observed before the occurrence of HCC, and the frequency of their appearance correlated well with the incidence of HCC. HBV DNA and the protein it encodes were detected in the cancer cells and/or the surrounding hepatocytes. Integration of HBV DNA inot the host liver genome was found during hepatocarcinogenesis among the animals infected by HBV. These results suggest that exposure to HBV and AFB1 may play a synergistic role in the development of HCC, and support the viewpoint of an aetiological relationship between HBV and HCC.Abbreviations HCC hepatocellular carcinoma - HBV human hepatitis B virus - AFB1 aflatoxin B1 - GGT foci hyperplastic hepatocyte foci positive for -glutamyltranspeptidase - LCD liver cell dysplasia     

Prevention of hepatocellular carcinoma in chronic viral hepatitis B and C infection

Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality worldwide,with the majority of cases associated with persistent infection from hepatitis B virus(HBV)or hepatitis C virus(HCV).Natural history studies have identified risk factors associated with HCC development among chronic HBV and HCV infection.High-risk infected individuals can now be identified by the usage of risk predictive scores.Vaccination plays a central role in the prevention of HBV-related HCC.Treatment of chronic HBV infection,especially by nucleoside analogue therapy,could also reduce the risk of HBV-related HCC.Concerning HCV infection,besides the advocation of universal precautions to reduce the rate of infection,pegylated interferon and ribavirin could also reduce the risk of HCV-related HCC among those achieving a sustained virologic response.Recently there has been mounting evidence on the role of chemopreventive agents in reducing HBV-and HCV-related HCC.The continued advances in the understanding of the molecular pathogenesis of HCC would hold promise in preventing this highly lethal cancer.     

HBx致肝细胞癌分子机制的研究进展

乙型肝炎病毒(HBV)慢性感染是我国肝细胞肝癌(HCC)发生的主要原因之一.一些体内外研究发现,乙型肝炎病毒x(HBx)可诱导肝细胞的恶性转化及癌变,成为目前研究乙肝相关性肝细胞癌发生机制的热点.近来在HBx与抑癌基因,如p53的新成员p73以及p16,p21等之间的相互作用及其对肝细胞恶性转化与癌变的影响等方面的研究也取得初步成果,但其关系错综复杂,这方面不断的深入研究将有助于进一步揭示HBx致肝细胞癌发生的分子机制,对探寻乙肝相关性肝癌新的防治策略具有重要意义.     

Clinical features of hepatitis B virus-related hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a major cause of cancer death,and chronic hepatitis B is a serious worldwide problem.The epidemiology of HCC is distinctive.Hepatitis B virus (HBV) plays a major role in hepatocarcinogenesis.Prevention of HBV-related HCC is a key issue in current hepatology.This paper describes the prevention and clinical features of HBVrelated HCC,along with a short review of the disease.     

Genotypes, mutations, and viral load of hepatitis B virus and the risk of hepatocellular carcinoma: HBV properties and hepatocarcinogenesis

Chronic infection with hepatitis B virus (HBV) is the major risk factor for hepatocellular carcinoma (HCC) worldwide. Ten HBV genotypes (A-J) have been discovered so far. Genotypes B and C are endemic in East and Southeast Asia. Genotype C HBV is associated with increased risks of cirrhosis and HCC. Genotype B (B2) is associated with the development of HCC in non-cirrhotic patients younger than 50 years and with relapse of HCC after surgical treatment. It is also associated with earlier hepatitis B e antigen seroconversion than genotype C. High HBV load is independently associated with the occurrence and post-treatment recurrence of HCC. Different genotypes have distinct patterns of mutations. Viral mutations in the core promoter region and in the preS region are frequently found to be significantly associated with an increased risk of HCC. These mutations often occur before the onset of HCC and accumulate during the progression of chronic HBV infection. Multiple such mutations are more frequent in patients with HCC and are specific for HCC. HBV subgenotypes, viral mutations, and viral load can be used for the prediction of HCC. Early identification of HBV-infected individuals who will eventually develop HCC will help to develop active prophylactic protocols to reduce or delay the occurrence of HCC.