A precore-defective mutant of hepatitis B virus associated with e antigen-negative chronic liver disease

The pathogenesis of chronic liver disease (CLD) due to persistent hepatitis B virus (HBV) infection has not been defined, but the disease activity is believed to correlate with the presence of hepatitis B e-antigen (HBeAg) antigenemia and high viremia. The molecular characterization of an HBV mutant isolated from an HBeAg-negative patient with severe CLD required amplification of the circulating HBV DNA (2 pg/ml) by the polymerase chain reaction (PCR). Direct sequencing of the nucleotides from five independent amplifications of the conserved precore region consistently revealed a G to A mutation in each of the two terminal codons of the precore region. Codon 28 was mutated from tryptophan-encoding TGG to a translational stop codon, TAG; codon 29 preceding the core initiation codon was changed from GGC to GAC. For biologic evaluation of these mutations on HBV replication and expression of HBeAg in vitro, HepG2 cells were transfected with cloned, recircularized mutant HBV DNA. The transfected cells contained subviral core particles in the cytoplasm and secreted mature HBV, without HBeAg, into the medium. The findings present the first evidence that complete HBV genomes can be amplified by PCR and are replication-competent in vitro. The data also indicate that HBeAg is not necessary for replication of HBV and furthermore suggest that HBeAg is not required for the progression of HBV-induced CLD.     

Dose-dependent increase in plasma interleukin-6 after recombinant tumour necrosis factor infusion in humans.

Several studies have shown that the cytokine interleukin-6 (IL-6) is produced in response to tumour necrosis factor (TNF) in vitro. This study examines the in vivo relation between these two cytokines with assays of plasma IL-6 and TNF levels in subjects with chronic hepatitis B undergoing immunomodulatory therapy with recombinant TNF (rTNF). Plasma IL-6 was detected from 20 min after rTNF infusion with levels peaking after 2-3 h and levels correlated with the dose of rTNF administered (r = 0.67, P = 0.004). Peak levels of IL-6 (mean 295, range 266-297 ng/l) were lower than those seen in certain disease states despite the very high peak levels of rTNF (mean 11,750, range 5623-18,620 ng/l). These findings suggest that the very high levels of IL-6 found in certain disease states are not purely the result of circulating TNF. Other factors such as endotoxin or other cytokines may also play a role in determining levels of plasma IL-6.     

A cDNA clone encoding a peptide highly specific for hepatitis C infection

A random primed lambda gtll-cDNA library was constructed from donors plasma presumably infected by blood-borne non-A, non-B hepatitis (hepatitis C:HC) agent and immunoscreened with serum pooled from patients with acute or chronic HC. Twelve lambda gtll-cDNA clones encoding antigens associated with HC infection in Japan as well as in the USA were isolated. Of these one clone consisting of 114 nucleotides and showing a discrete band on an immunoblot analysis, was extensively studied. The clone is not derived from the host DNA encoding one polypeptide specific and highly sensitive for serum from patients with HC and has no homology to the nucleotide sequences of known human viruses including hepatitis A, B and D viruses, Ebstein-Barr virus, coxsackievirus, immunodeficiency virus type 1 or Japanese encephalitis virus. These results suggest that this clone is derived from the genome of HC agent.     

慢性乙型肝炎肝内HBV DNA和HBsAg的双标记定位研究

应用原位杂交和免疫组化PAP法双标记技术,结合病人乙型肝炎(乙肝)病毒血清学标志物检测结果,研究了31例慢性乙肝病人肝穿刺组织中乙型肝炎病毒DNA(HBV DNA)和HBsAg的分布及意义。结果显示,肝组织内检出HBV DNA 23例,HBsAg 26例,二者同时检出者21例。从同组病人肝组织内HBV DNA和HBsAg双标检测结果与其乙肝病毒血清学标志物检测结果的比较来看,肝组织内HBV DNA和HBsAg同时阳性很可能表明HBV正处于复制及表达的活动状态。     

重型乙型肝炎患者外周血中IL-2系统的初步研究

以放射免疫分析法、微量细胞培养法检测31例重型乙型肝炎患者血清和用PHA-P、rIL-2刺激外周血PBMC的培养上清中IL-2、sIL-2R含量,以间接免疫荧光法检测培养后PBMC中Tac~+细胞数,结果显示:重型乙型肝炎患者血清、培养上清中sIL-2R含量和Tac~+细胞数显著高于慢性活动型肝炎、急性黄疸型肝炎及献血员对照组,而血清、上清中IL-2含量与急性肝炎接近,但显著高于慢性活动性肝炎患者(P<0.01)。提示:重型乙型肝炎患者IL-2R表达增强,PBMC处于较高的免疫活化状态,其IL-2系统的生物学活性有可能高于各对照组。     

Detection of IgA class antibody to hepatitis E virus in serum samples from patients with hepatitis E virus infection

A newly developed assay for IgA class antibody to hepatitis E virus (IgA anti-HEV) was used to study 145 serum samples collected during an outbreak of an enterically transmitted hepatitis that occurred in 3 villages in the lower Shebeli region of Southern Somalia between January, 1988 and November, 1989. A total of 52.4% of the afflicted patients were found positive for IgA anti-HEV, and 73.1% of these were also positive for IgM. Both antibodies disappeared during the convalescence period. Similar results were also seen in serum obtained from sporadic cases of acute waterborne hepatitis in Pakistan. © 1993 Wiley-Liss, Inc.     

丹红注射液联合甘利欣治疗慢性乙型肝炎肝纤维化的临床研究

目的探讨丹红注射液联合甘利欣注射液治疗慢性乙型肝炎肝纤维化的临床疗效。方法选取192例慢性乙型肝炎肝纤维化患者,随机分为治疗组110例,对照组82例,治疗组用丹红注射液联合甘利欣注射液治疗,对照组单用甘利欣注射液。观察疗程结束前后患者血清透明质酸（HA）、血清Ⅲ型前胶原（PC-Ⅲ）、层粘蛋白（LN）、Ⅳ型胶原（Ⅳ-C）、肝功能指标变化及临床疗效。结果治疗组血清肝纤维化指标、肝功能指标下降明显,而白蛋白有明显升高,与对照组相比有显著性差异（P〈0.05或P〈0.01）。结论丹红注射液联合甘利欣注射液治疗慢性乙型肝炎,可有效减轻和延缓肝纤维化的发生与发展,对肝纤维化指标和肝功能有明显改善作用。     

不同基因型HBx对乙型肝炎病毒复制的影响

目的探讨A、B、C和D四个基因型乙型肝炎病毒X蛋白（hepatitis Bvirus Xprotein,HBx）对乙型肝炎病毒（hepatitis Bvirus,HBV）复制的影响。方法构建含A、B、C和D四种基因型HBx的真核表达载体和提前终止HBx表达的突变型单倍体HBV表达系统,共转染至人肝肿瘤细胞系HepG2细胞,利用荧光素酶报告系统检测不同基因型HBx对HBV核心启动子的激活作用,利用酶联免疫法（ELISA）检测培养上清中HBV表面抗原（HBsAg）和e抗原（HBeAg）的表达情况,利用特异性引物PCR方法检测细胞中HBV前基因组RNA（pgRNA）和总RNA（pgRNA,2.2kb和0.8kbRNA）的转录情况。结果 A、B、C和D四种基因型HBx的表达均可显著激活HBV核心启动子活性,但不同基因型HBx的激活作用不同。除C基因型外,A、B和D三种基因型HBx均可显著增强HBsAg和HBeAg的胞外分泌水平。检测HBV复制中间产物发现,C基因型HBx对HBVpgRNA和总RNA转录水平的影响显著低于A、B和D基因型。结论不同基因型HBx对HBV复制的影响不同,为进一步研究HBV基因型与HBV复制能力以及临床病理表现的关系打下基础。     

胶体金法检测乙肝病毒血清标志物的实验研究

 目的:客观评价胶体金法检测乙肝病毒标志物(HBVM)的准确性,了解胶体金法与ELISA法之间的差异.方法:收集急诊手术患者的血清标本1100份,用胶体金法和ELISA法同时检测,以ELISA方法检测结果为参照,比较胶体金法检测HBVM的灵敏度和特异性.结果:与ELISA胶体金法为标准计算HBsAg和HBeAg的灵敏度和特异性均超过90%;其他3个抗体的灵敏度和特异性均低于80%.结论:胶体金法检测HBsAg和HBeAg适用于急诊筛检,乙肝三项抗体金标法存在较严重的漏检,不适合急诊筛查.     

广东省62例自身免疫性肝炎的流行病学及临床特征分析

目的 研究自身免疫性肝炎(AIH)的流行病学及临床特征.方法 收集2002年11月1日-2014年3月31日在暨南大学附属第一医院诊断为AIH患者的住院资料,分析其流行病学、临床特征、免疫球蛋白IgG或γ球蛋白水平、肝功能及肝纤维化指标.结果 ①共纳入62个病例,其中男9例,女53例,男女比例1∶5.89,患者年龄15～79岁,中位年龄56岁.62例患者中,中年患者占40.3％(25例);乏力(61.3％)、肝大(54.8％)、纳差(53.2％)为最常见症状;Ⅰ型AIH占77.42％,Ⅱ型3.23％,Ⅲ型3.23％,Ⅳ型16.13％.②不同分型AIH患者性别、年龄构成比、临床表现及并发症发生率、血清IgG或γ球蛋白水平和肝功能指标之间的差异均无统计学意义.③Ⅰ型AIH患者男性腹胀、肝掌、蜘蛛痣、双下肢水肿、上消化道出血、自发性腹膜炎发生率均高于女性,而女性血清谷氨酰转肽酶(GGT)、碱性磷酸酶(ALP)浓度高于男性,血清透明质酸(HA)及层粘连蛋白(LN)浓度低于男性.Ⅰ型AIH患者各年龄组临床表现及并发症发生率、血清IgG或γ球蛋白水平、肝功能指标差异均无统计学意义.结论 AIH以中年女性多见,以乏力、肝大、纳差为最常见症状,Ⅰ型AIH最多,女性患者血清GGT、ALP浓度明显高于男性,而血清HA及LN浓度则低于男性.