原位杂交检测乙肝病毒携带者精子中乙肝病毒-DNA的研究

目的通过检测乙肝病毒携带者经辅助生育技术洗涤后的精子是否携带有乙型肝炎病毒,从而判断是否存在经精子传播乙型肝炎的可能性。方法用地高辛标记,针对乙型肝炎病毒DNA(HBV-DNA)设计的探针,对来我院要求行辅助生育治疗的20例乙肝病毒携带者的精子,经洗涤离心上游后进行检测。结果发现8例携带者精子中有乙肝病毒DNA,分布于精子的头部,阳性精子计数占每片精子总数的3‰～10‰。结论经辅助生育技术洗涤后的乙肝病毒携带者的精子仍可传播HBV-DNA。     

The existence and significance of hepatitis B virus DNA in glomerulonephritis

Summary: In order to investigate the role of hepatitis B virus (HBV) in the pathogenesis of glomerulonephritis 50 cases of glomerulonephritis with HBV antigenaemia and/or hepatitis B antigen (HBAg) detected by immunohistochemistry in renal tissue were collected. the distribution and localization of HBV DNA were observed by using in situ hybridization. In addition, Southern blot analysis was performed on 23 of the 50 cases in order to reveal the state of renal HBV DNA. Thirty-six cases (72%) were found to be HBV DNA positive by in situ hybridization, which localized in the nucleus of tubular cells. In 26 cases HBV DNA was detectable in the nucleus of glomerular mesangial and epithelial cells as well as the mesangial matrix. Seventeen of the 23 cases were proved to be HBV DNA positive in Southern blot analysis (82%). Three of these cases were identified with non-replicating free HBV DNA, while 14 cases were the integrated form. the results of this study showed that the renal tissue was infected with HBV; however, it was considered that it may be possible that the HBAg deposited on glomeruli was not only from circulation but also from the HBV infected glomerular cells although the evidence of this is not conclusive. In addition to the humoral immune injury mediated by HBAg-hepatitis B antibody (HBAb) immune complexes the cellular immune injury mediated by target antigen (hepatitis B core antigen; HBcAg) might be also involved in the pathogenesis of HBV glomerulonephritis (GN) associated GN.     

乙型肝炎病毒相关性肾炎发病机制的研究进展

乙型肝炎病毒相关性肾炎(HBV-GN)是指由乙型肝炎病毒感染后引起的肾小球肾炎,是我国常见的继发性肾小球肾炎之一,多发于儿童及中青年人,男性多见。HBV-GN的发病机制目前尚不清楚,可能与病毒、宿主及环境因素的相互作用有关。近年来,关于乙型肝炎病毒基因及其其他分子生物学方面的研究取得了一些重要进展,本文将对此进行综述,以期对HBV-GN诊断和治疗提供新的方向和理论依据。     

The effect of hepatitis B vaccination on the incidence of childhood HBV-associated nephritis

The aim of the present study was to investigate the effect of a vaccination program for hepatitis B virus (HBV) on the incidence of HBV-associated glomerulonephritis (HBV-GN). In total, 727 renal biopsies were carried out at our hospital from November 1979 through March 2002. Two groups were established. Group A included those biopsied from November 1979 through December 1991 (prior to the HBV vaccination program) and group B from January 1992 through March 2002. Group B was divided into five subgroups (B₁ to B₅), with an interval of 2 years between each subgroup. Patients were divided into those with or without a history of HBV vaccination. Of the 727 renal biopsies, 64 fulfilled the criteria of HBV-GN, There were 28 cases of the 211 cases in group A and 36 cases of the 516 cases in group B (X ²=7.397,P<0.01). The incidence in group A and group B₁ through B₅ was 13.27% (28/211), 13.04% (9/69), 7.32 (6/82), 6.25% (4/64), 4.88% (4/82), and 5.94% (13/219), respectively (X ²=9.627, P<0.01). Only 8 of the 231 vaccinated children had HBV-GN, while there were 48 HBV-GN cases of the 381 non-vaccinated children (X ²=14.44, P<0.001). There were only 6 cases of membranous nephropathy (MN) in the vaccinated group, while 40 cases of MN occurred in the non-vaccinated group (X ²=12.92, P<0.01). There were 8 children that developed HBV-GN with abnormal serum HBV markers despite HBV vaccination. Two mothers of these 8 children had evidence of HBV infection. The incidence of HBV-GN in children has been decreasing each year since the implementation of the nationwide HBV vaccination program in Shanghai, China. Furthermore, since childhood MN is associated with HBV, vaccination can also reduce the incidence of childhood MN.     

Absence of hepatitis B and C viruses in pediatric idiopathic membranoproliferative glomerulonephritis

The blood-borne hepatitis viruses, hepatitis B virus (HBV) and hepatitis C virus (HCV), have similar epidemiological features. The association of chronic HBV infection and glomerulonephritis is well established, particularly in children. Recent reports have shown an association between HCV infection and glomerulonephritis in adults. In order to assess the role of these hepatotropic viruses in membranoproliferative glomerulonephritis (MPGN) we screened 34 children with idiopathic MPGN for the presence of HBV and HCV infection using highly sensitive polymerase chain reaction techniques for the detection of HBV DNA and HCV RNA. Also, enzyme-linked immunosorbent assays were used to detect the presence of antibody to hepatitis B surface antigen and antibody to HCV. No evidence of HBV or HCV infection was demonstrated in any of the patients. We conclude that HBV and HCV are not significant causes of idiopathic MPGN in children in the United States.     

乙型肝炎病毒相关性肾炎的相关因素分析

目的 研究乙型肝炎病毒相关性肾炎（HBV-GN）的相关因素。 方法 采用回顾性病例对照研究。用Logistic回归对86例HBV-GN患者及135例非HBV-GN慢性HBV感染者进行比较。 结果 单因素分析结果显示,HBV-GN与男性（OR 2.79,95％CI 1.48～5.25,P = 0.001）、HBeAg阳性（OR 2.60,95％CI 1.49～4.53,P = 0.001）、HBV复制（OR 3.63,95％CI 1.80～7.33,P < 0.01）、肝硬化（OR 4.58,95％CI 1.41～14.91,P = 0.011)及谷丙转氨酶（ALT）升高（OR 2.53,95％CI 1.42～4.51,P = 0.002)相关。多因素分析结果显示,HBV-GN与男性（OR 2.21,95%CI 1.12～4.33,P = 0.022）、HBV复制（OR 2.77,95%CI 1.28～5.97,P = 0.01）、肝硬化（OR 4.55,95%CI 1.29～16.10,P = 0.019）及ALT升高（OR 1.96,95%CI 1.04～3.69,P = 0.037）相关。HBV-GN中,与IgA肾病型（HBV-IgAN）比较,单因素分析结果显示,经典型HBV-GN（膜性肾病或膜增生性肾炎型）与男性（OR 5.24,95％CI 1.64～16.81,P = 0.005）、HBV复制（OR 8.02,95％CI 2.15～29.84,P = 0.002）及ALT升高（OR 3.10,95％CI 1.00～9.67,P = 0.051）相关;多因素分析结果显示,仅与男性（OR 6.51,95%CI 1.76～24.11,P = 0.005）及HBV复制（OR 7.22,95%CI 1.68～30.97,P = 0.008）相关。结论 男性、HBV复制、肝硬化及ALT升高都可能是HBV-GN的易患因素。与HBV-IgAN比较,经典型HBV-GN与病毒复制及男性更相关,两者可能有不同的病理机制。     

乙型肝炎病毒相关性肾炎的诊治——附20例报告

目的总结乙型肝炎病毒相关性肾炎（HBV-GN）患者的临床及病理特点,以提高HBV-GN的诊治水平。方法经肾活检确诊为HBV-GN的患者20例,回顾性分析患者临床表现、实验室检查（尿常规、24h尿蛋白定量、转氨酶、血清清蛋白、HBV-DNA等）、肾组织光镜改变及免疫荧光（IgG、IgA、IgM、C3、C1q、FRA、HBsAg、HBcAg、HBeAg）染色结果,并分析其临床及病理特点。结果本组20例,其中肾病综合征10例,肾炎综合征9例,无症状镜下血尿1例。病理分型以非典型膜性肾病最多见（MN）,共16例（占78％）,系膜增生性肾小球肾炎（MsPGN）2例（占10％）,系膜毛细血管性肾炎（MPGN）2例（占10％）。7例患者HBV-DNA阴性。给予抗病毒联合激素治疗后,大多数患者肾功能稳定。结论目前HBV-GN的确诊仍依赖肾活检,在乙型肝炎高发区应常规进行肾活检组织乙型肝炎病毒标志物检测。     

Resolution of hepatitis B virus-related membranoproliferative glomerulonephritis after orthotopic liver transplantation

The most well-described renal disease associated with hepatitis B virus (HBV) infection is membranous glomerulonephritis; membranoproliferative glomerulonephritis is described much less frequently. The course of HBV-associated renal disease after liver transplantation has not been described to date. We present a 15-year-old girl with HBV-associated membranoproliferative glomerulonephritis and end-stage liver disease, in whom, after cadaver liver transplantation, clinical and histological resolution of renal disease was observed. Resolution was associated with diminution of circulating HBV surface antigen levels.     

Interferon treatment for hepatitis B-associated membranous glomerulonephritis in two Chinese children

Two Chinese boys, aged 3.5 and 5 years, developed nephrotic syndrome and were chronic carriers of hepatitis B virus surface antigen (HBsAg) and hepatitis B virus e antigen (HBeAg). Renal biopsy showed membranous glomerulonephritis and liver biopsy showed chronic persistent hepatitis. They were given interferon--2a at a dose of 5 MU/m² on alternate days for 12 and 16 weeks after 2 years of persistent nephrotic syndrome. Patient 1 showed complete remission and resolution of hepatosplenomegaly, but his serum remained positive for HBsAg, HBeAg and hepatitis B virus DNA. Patient 2 showed only a transient clinical response and seroconversion from HBeAg to anti-HBe status. Although not always successful, interferon treatment should be considered in severe persistent nephrotic states, since there is at present no satisfactory treatment for this form of glomerulonephropathy.     

肝外胆管癌组织中HBx mRNA的原位检测和临床病理意义

目的 检测肝外胆管癌和癌旁胆管组织中乙型肝炎病毒X基因(HBx)mRNA的表达,了解HBV感染与胆管癌发生间的关系,明确HBx蛋白和基因在胆管癌发生中的意义。方法酶切质粒凝胶回收HBx基因片段,地高辛标记HBx DNA探针,原位杂交检测胆管癌及癌旁胆管石蜡标本中HBx mRNA的表达,并与临床病理资料进行相关分析。结果71例肝外胆管癌组织中HBx mRNA表达阳性率为61％(43／71);而39例癌旁胆管组织中的HBx mRNA表达阳性率仅为18％(7／39),且阳性信号全部见于伴有中重度不典型增生的胆管上皮细胞内。统计分析显示胆管癌组织中HBx mRNA表达和分布与临床病理参数无相关性,但HBx蛋白和基因的表达呈正相关。结论 肝外胆管癌组织中可以检测到HBx mRNA表达,HBV感染及其基因整合在胆管癌发生中有一定意义。     

间质性肾炎中EB病毒的原位杂交检测

明确EB病毒(EBV)在间质性肾炎患者肾组织中的感染部位。方法 用原位杂交方法对12例间质性肾炎患者及10例微小病变肾病患者对照组肾组织石蜡切片进行EB病毒编码的小RNA(EBER1)检测。结果 发现3例阳性,主要分布于肾小管上皮细胞,间质浸润细胞及部分肾小球内的细胞胞核中,其中2例为特发性间质性肾炎,1例为药物过敏性间质性肾炎,对照组均为阴性。结论 支持EB病毒感染在间质性肾炎发病中具有重要作用的推测,在不同类型的间质性肾炎中,EB病毒感染的作用不尽相同,不应一概而论。     

Interferon treatment on glomerulonephritis associated with hepatitis C virus

We report on a 10-year-old girl with glomerulonephritis associated with hepatitis C virus infection, who was treated with interferon-α. On the first renal biopsy at 8 years of age, mild mesangial hypercellularity in a segmental to semiglobal pattern was present in all glomeruli. After 6 months interferon-α therapy, proteinuria diminished completely. However, mesangial proliferation was advanced on the second biopsy at 10 years of age. We concluded that the interferon-α was effective in the treatment of proteinuria despite the lack of pathological improvement. Received: 15 March 2000 / Revised: 11 July 2000 / Accepted: 14 July 2000     

血清HBsAg、HBcrAg和HBV DNA预测慢性乙型肝炎肝脏病理状态的评价

目的评价血清乙型肝炎病毒表面抗原（HBsAg）、乙型肝炎病毒核心相关抗原（HBcrAg）和乙型肝炎病毒DNA（HBV DNA）预测慢性乙型肝炎（CHB）患者肝脏不同病理学分级和分期的有效性。 方法共选取CHB患者211例,其中HBeAg阳性和阴性分别为125例和86例。血清HBsAg、HBeAg和HBcrAg分别采用化学发光微粒子免疫法和化学发光酶免疫法检测,血清HBV DNA采用实时荧光定量PCR检测。 结果HBeAg阳性患者,血清HBsAg、HBcrAg和HBV DNA与肝脏病理学分级和分期均呈显著负相关（P均< 0.05）;HBeAg阴性患者,血清HBcrAg和HBV DNA与肝脏病理学分级和分期均呈显著正相关（P均< 0.01）。Bayes逐步判别分析显示,HBeAg阳性患者,仅血清HBsAg符合判别肝脏不同病理学分级和分期的方程纳入自变量的标准（F > 3.84）,Fisher判别函数预测肝脏病理学分级G1、G3和分期S1、S4的正确率分别为62.7%、58.8%和63.8%、69.6%;HBeAg阴性患者,血清HBV DNA和HBcrAg分别符合判别肝脏不同病理学分级和分期的方程纳入自变量的标准（F > 3.84）,Fisher判别函数预测肝脏病理学分级G1、G3和分期S1、S4的正确率分别为79.3%、66.7%和71.7%、75.0%。 结论血清HBsAg对HBeAg阳性患者以及血清HBV DNA和HBcrAg分别对HBeAg阴性患者的肝脏病理学分级G1、G3和分期S1、S4有显著的预测意义。     

Efficacy of adefovir dipivoxil combined with a corticosteroid in 38 cases of nephrotic syndrome induced by hepatitis B virus-associated glomerulonephritis

Objective: To investigate the treatment efficacy of adefovir dipivoxil combined with a corticosteroid on hepatitis B virus-associated glomerulonephritis (HBV-GN). Methods: A total of 38 patients with nephrotic syndrome induced by HBV-GN were treated for 36 weeks between 2010 and 2012. Results: The efficacy analysis showed that 11 patients achieved complete remission and 17 patients achieved partial remission, and the effective remission rate was 73.7%. In addition, 10 patients achieved no remission. Conclusions: Adefovir dipivoxil combined with corticosteroids has a certain efficacy on the HBV-GN and displays few adverse reactions. A large sample, randomized double-blind controlled study and long-term follow-up are needed to verify the efficacy of adefovir dipivoxil combined with corticosteroids.     

血清 HBV DNA 定量与 HBV 相关性肾炎患者临床病理特征及预后的相关性

目的：探讨HBV相关性肾炎（ HBV-GN）患者血清HBV DNA复制水平与临床病理特征及预后的相关性。方法选取2009年1月至2012年10月在青岛大学附属医院接受治疗的HBV-GN患者102例。采用荧光定量聚合酶链反应法测定所有患者血清HBV DNA载量,并依此将患者分为HBV DNA低度复制组（ HBV DNA＜103拷贝／mL）、HBV DNA中度复制组（103拷贝／mL≤HBV DNA≤105拷贝／mL）和HBV DNA高度复制组（ HBV DNA﹥105拷贝／mL）。所有患者行肾脏穿刺活检,明确病理类型,并进行肾脏组织中HBV相关抗原（ HBsAg、HBcAg及HBeAg）的免疫荧光定量检测。所有患者均接受拉米夫定（100 mg／d）联合阿德福韦酯（10 mg／d）治疗,随访观察18个月,并对各组患者治疗前后的肾功能和生化免疫学指标进行比较。采用One-way ANOVA单因素方差分析比较不同HBV DNA复制水平的患者肾功能、生化免疫学指标、肾脏组织中HBV相关抗原沉积的差异,并采用Spearman相关分析法分析血清HBV DNA水平与肾脏病理分期的相关性。结果102例患者中, HBV DNA低度复制组20例,HBV DNA中度复制组51例,HBV DNA高度复制组31例。随着血清HBV DNA复制水平的增高,患者24 h尿蛋白定量、胆固醇及三酰甘油呈现不同程度的增加（ F＝34.64,40.10和31.72,P＜0.01）,血浆白蛋白降低（F＝24.04,P ＜0.01）;肾脏组织中 HBV 抗原（HBsAg、HBcAg及HBeAg）免疫复合物的沉积增加（F＝41.49,15.64和10.41,P＜0.01）。对78例膜性肾病（MN）患者分析发现,随着血清HBV DNA复制的增加,HBV-MN患者病理分期加重（r＝0.38, P＜0.01）。抗病毒治疗后,各组24 h尿蛋白排泄量较治疗前均显著下降（ t＝7.86,19.28和16.74, P＜0.01）;补体C3较治疗前上升,但与治疗前比较差异无统计学意义（t＝1.05,1.04和1.94,P＞0.05）;血肌酐较治疗前无明显变化（ t＝0.14,0.52和0.57,P＞0.05）。治?     

乙型肝炎病毒相关性肾炎的中西医结合治疗

乙型肝炎病毒相关性肾炎(HBV-GN)的研究已40余年。其发病机制仍不十分明确,可能与病毒介导的免疫反应有关。HBV－GN诊断标准为：(1)血清HBV抗原阳性;(2)确诊为肾小球肾炎;(3)肾组织中找到HBV抗原,其中(2),(3)为必要条件。在诊断HBV-GN时应注意排除肾组织HBV标志物假阳性;排除HBV肾组织沉积伴发原发性肾炎;注意与狼疮肾炎的鉴别。HBV-GN的肾脏病理以膜性肾病及膜增生性肾炎常见。HBV-GN以肾病综合征及轻微的血尿为主要临床表现。儿童HBV-GN患者的病程多呈自限性,而成人HBV-GN则常呈缓慢进展,常发展为慢性肾功能衰竭。关于HBV-GN治疗的临床报道很多,多为无对照或小样本的回顾性研究,尚无统一的治疗原则可循。抗病毒治疗对HBV-GN有一定疗效,而激素和免疫抑制剂的使用尚存争议。抗病毒药[干扰素或核苷类]＋免疫抑制剂[激素和(或)其他免疫抑制剂]＋中药(辨证施治)的治疗模式,可能是较好的选择。     

HBx基因在乙型肝炎病毒相关性肾炎致病机制中的研究进展

HBx基因是一种多功能的调节因子,具有广泛的反式激活作用,能激活转录因子、抑制DNA修复,并调节细胞的增殖、转化及凋亡.近年来,HBx在乙型肝炎病毒相关性肾炎（HBV-GN）致病中的作用成为研究的热点.研究显示,HBx对肾小球系膜细胞增殖、足细胞及肾小管上皮细胞的损伤及凋亡存在广泛影响.此文就HBx的生物学特性及在肾脏损伤机制中的研究进展进行了综述.     

乙型肝炎病毒X蛋白对DNA甲基转移酶3A/3B表达的影响

目的观察乙型肝炎病毒X蛋白（HBx）对QSG7701细胞中DNA甲基转移酶（DNMT）3A/3B表达的影响。方法本研究室前期构建好的重组表达质粒pcDNA-X及空载体pcDNA3.0分别转染QSG7701细胞,经含G418的选择性培养基筛选获得稳定转染HBV-X基因的细胞克隆（pcDNA-X/QSG7701）及稳定转染空载体pcDNA3.0的细胞克隆（pcDNA3.0/QSG7701）。采用RT-PCR、Westernblot分别检测转染细胞中HBxmRNA和HBx蛋白的表达。Real-timePCR检测3种细胞DNMT3A/3BmRNA的表达情况。免疫组织化学法检测3种细胞DNMT3A/3B蛋白的表达情况。结果RT-PCR、Westernblot检测结果显示pcDNA-X/QSG7701细胞中有HBxmRNA及HBx蛋白的表达。Real-timePCR结果显示pcDNA-X/QSG7701中DNMT3A/3BmRNA表达水平显著高于pcDNA3.0/QSG7701及未转染的细胞QSG7701（P〈0.05）。免疫组织化学检测结果显示pcDNA-X/QSG7701细胞中DNMT3A/3B蛋白表达水平显著高于pcDNA3.0/QSG7701及QSG7701细胞（P〈0.05）。结论稳定转染HBV-X基因的人源性永生化非瘤性肝细胞QSG7701中DNMT3A/3BmRNA和蛋白的表达水平均显著升高,提示HBV-X基因在mRNA及蛋白水平能上调转染细胞中DNMT3A/3B的表达,而细胞中DNMT3A/3B表达的增加是否能进一步影响癌基因、抑癌基因的表达水平,从而导致细胞癌变,尚有待进一步研究。     

HBeAg阳性慢性乙型肝炎患者HBV前C/BCP突变/准种与HBeAg和HBV DNA的关系

目的探讨HBeAg阳性慢性乙型肝炎（eP-CHB）HBV前C/BCP突变/准种及其与HBeAg、HBV DNA水平的关系。 方法采用断面研究对2016年1月至2018年12月就诊于首都医科大学附属北京佑安医院的220例eP-CHB患者进行前C/BCP突变检测，其中24例患者进行前C/BCP区扩增、克隆，同步检测血清HBeAg和HBV DNA水平，分析前C/BCP突变/准种的发生情况及其与HBeAg和HBV DNA水平的关系。 结果220例eP-CHB患者中，HBV前C/BCP总突变率为70.0%（154/220），前C/BCP共同突变率为18.2%（40/220），前C突变率为30.9%（68/220），BCP突变率为57.3%（126/220）。HBV DNA≥ 5 lgIU/ml患者上述4种突变检出率均高于HBV DNA< 5 lgIU/ml者，其中前C/BCP总突变和BCP突变患者差异有统计学意义（χ² = 5.809、P = 0.016，χ² = 5.081、P = 0.024）。HBeAg水平越低（< 500 COI、500～1 000 COI和> 1 000 COI共3组患者比较），以上4种突变检出率越高，差异有统计学意义（χ² = 31.738、17.291、16.263、22.164，P均< 0.001）。HBV DNA≥ 5 lgIU/ml患者中，HBeAg水平越低，以上4种突变检出率越高，差异亦均有统计学意义（χ² = 40.503、19.654、16.727、29.119，P < 0.001）。准种检测中，前C区高突变组患者HBeAg水平低于低突变组，差异有统计学意义（t = 2.230、P = 0.017），前C、BCP高突变组与低突变组间HBV DNA水平差异无统计学意义（t = 0.624、P = 0.462，t = 0.893、P = 0.317）。 结论eP-CHB患者中仍存在广泛的前C/BCP突变。高HBV DNA、低HBeAg表达者，前C和BCP突变的发生率较高；前C区突变株在准种中比率高者更影响HBeAg的表达。推测前C/BCP突变可能是eP-CHB出现低HBeAg、高HBV DNA，并导致抗病毒治疗停药后易复发的原因。