IgA肾病肾组织内乙型肝炎病毒感染的发病机制研究

目的：探讨乙型肝炎病毒感染致IgA肾病肾损伤的发病机制。方法： 随机选取48例IgA肾病肾穿刺组织，参照Meadow病变分级标准分为Ⅰ-Ⅴ级5个实验组，应用Envision免疫组织化学方法检测各级肾组织内HBsAg和HBcAg；同时用直接IS-PCR技术检测其中18例IgA肾病肾组织内HBV DNA。结果： 48例IgA肾病肾组织内HBcAg和HBsAg总的阳性检出率分别为75.00%(36/48)和43.75%(21/48)；18例IgA肾病肾组织内HBV DNA阳性检出率为61.11%(11/18)；3者均表现为肾小管阳性检出率高于肾小球(P＜0.05)，但各级之间，HBcAg、HBsAg和HBV DNA检出率均无显著差异(P＞0.05)。结论： HBV参与了IgA肾病的发生，其导致肾组织损伤的机制可能主要是由细胞免疫或一系列细胞因子介导，并非病毒直接所致；肾小管上皮细胞可能是HBV感染的靶对象。     

肾组织preS1/S2-Ag检测在乙型肝炎病毒相关性肾炎诊断中的作用

 目的：观察前S1/S2抗原（preS1/S2-Ag）及其它乙型肝炎病毒(HBV)抗原在HBV相关性肾小球肾炎（HBV-GN）患者肾组织中的表达,探讨它们在HBV-GN诊断中的应用价值。方法：回顾性收集2003年1月~2013年1月于中山大学附属第三医院肾内科住院的患者,并符合以下入组标准：血清学HBsAg阳性;有血尿、蛋白尿等临床表现;经肾组织活检病理诊断为肾小球肾炎病变;排除合并有系统性红斑狼疮、过敏性紫癜、糖尿病、丙肝等疾病。入组共49例。用免疫组织化学的方法对其肾组织石蜡切片进行preS1/S2-Ag、HBeAg、HBsAg和HBcAg检测。随机选取 5例HBsAg阴性的肾小球轻微病变患者,5例HBsAg阳性的非肾小球肾炎患者作对照。结果： 49例HBV感染合并肾小球肾炎患者中preS1/S2-Ag、HBeAg、HBsAg和HBcAg检出阳性率分别为32.7%（16例）、 388%（19例）、14.3%（7例）和46.9%（23例）,总阳性率为70.2%（36例）;preS1/S2-Ag主要表达在肾小管上皮细胞、肾小球系膜细胞和内皮细胞胞质内;其表达与肾组织HBcAg的表达呈正相关（r=0459, P<001）。5例HBsAg阴性的肾小球轻微病变患者,4种抗原均未检测到;5例HBsAg阳性的非肾小球肾炎患者有2例表达HBeAg,1例表达HBcAg,无1例表达preS1/S2-Ag及HBsAg。结论：HBV-GN患者肾组织中检测到preS1/S2-Ag; preS1/S2-Ag、HBeAg、HBsAg和HBcAg联合检测可提高HBV-GN的临床诊断率。     

血清HBV标志物阴性肾炎肾组织中HBV-DNA的原位杂交检测

目的 对血清HBV标志物阴性肾炎患者肾组织进行HBV—DNA的检测，为临床诊疗提供理论依据。方法 应用原位杂交（ISH）技术检测37份血清HBVAg阴性和18份血清HBVAg阳性的肾炎患者肾活检石蜡包埋组织切片中的HBV—DNA。结果 在血清HBVAg阳性的18例标本中HBV-DNA的检出率为88．9％（16／18），血清HBVAg阴性37例中检测出HBV—DNA5例（13．5％）。原位杂交显示两组检出的HBV-DNA均以肾小管上皮细胞质分布为主，与组织中HBVAg阳性颗粒的分布基本一致。结论 血清HBVAg阴性的HBV相关性肾炎临床上并非少见，应给予足够的关注。HBV—DNA在肾组织中的检出，表明HBV在HBV相关性肾炎的发病机理中有着重要作用。     

乙型肝炎病毒前S1蛋白抗原检测的临床意义

目的通过分析HBV前S1（PreS1）蛋白抗原和血清标志物之间的关系,探讨PreS1蛋白抗原检测在临床上的应用价值。 方法收集382例HBV患者血清样本,采用ELISA法检测HBVPreS1蛋白抗原和血清标志物,采用荧光定量PCR法检测HBV—DNA,并比较所有检测结果。 结果382例HBV患者血清样本PreS1蛋白抗原和HBV-DNA的检出率分别为33．8％和40．8％;其中HBeAg阳性的144例中,PreS1抗原阳性为112例,HBV-DNA阳性为134例,阳性率分别为77．8％和93．1％;HBeAg阴性的102例中,PreS1抗原阳性为17例,HBV-DNA阳性为22例,阳性率分别为7．1％和9．2％。HBV-DNA检出率稍高于PreS1抗原的检出率,但两者间检出率差异均无统计学意义（P〉0．05）。 结论HBVPreS1蛋白可反映HBV复制的情况,与HBV-DNA结果有较好的一致性,可作为临床上HBV感染诊断或监测的指标。     

HBV DNA及HBV抗原在血清HBV标志物阴性肝炎肝组织中表达的研究

目的 研究HBV DNA及HBV抗原在血清HBV标志阴性的肝炎肝组织中的表达。方法 对45例HBV血清标志阴性阴性肝炎患者，进行肝组织HBV DNA的原位杂交及免疫组织化学染色检测。结果 原位杂交表明，HBV DNA阳性者7例，（阳性率15．56％），阳性信号主要存在于肝细胞的胞核中，少数位于胞浆内；免疫组化染色表明，HBsAg及HBcAg均呈阴性。结论 血清HBV标志阴性的肝炎肝组织中可检出HBV DNA，有利于提高对HBV感染的诊断。     

HBV携带者血清病毒标志物和HBV DNA与肝组织中HBVcccDNA相关性的研究

目的探讨HBV携带者血清病毒标志物和HBV DNA与肝组织中HBVcccDNA之间的关系。方法应用实时荧光定量聚合酶链反应（RT-PCR）方法检测30例经肝组织活检病理检查确定为HBV携带者的肝组织中HBVcccDNA、HBVtDNA和血清HBV DNA,同时用化学发光免疫分析法检测HBsAg、HBeAg定量,分析感染者肝组织内HBVcccDNA与肝组织内HBVtDNA、血清HBVDNA、HBsAg及HBeAg定量水平之间的关系。结果HBV携带者肝组织中均可检出HBVcccDNA,范围在3．15×10^3拷贝／mg～1．06×10^7拷贝／mg（对数值：5．66±O．93）;肝组织cccDNA定量与肝组织总HBVDNA定量呈正相关（r=0．375,P〈0．05）,与血清HBVDNA无相关性（r=0．174,P〉0．05）。肝组织中HBVcccDNA水平与血清HBsAg定量呈高度正相关（r=0．562,P〈0．001）;而与血清HBeAg定量无相关性（r=0．152,P〉0．05）。结论HBV携带者肝组织内HBVcccDNA成稳定的中等水平复制;血清HBVDNA载量不能直接代表其肝组织中的HBVcccDNA水平;血清HBsAg定量可作为反映肝幺日织中HBVcccDNA水平的指标。     

HBsAg阳性母亲死胎肾脏组织中HBV表达

目的探讨乙型肝炎病毒通过产妇传播在胎儿肾脏组织中表达的情况。方法采用我院行中期妊娠引产,HBsAg阳性产妇分娩的死胎30例。取死胎肾脏组织用SP法检测死胎肾脏组织中HBcAg。结果孕妇血清乙型肝炎标志物小三阳及大三阳分娩的死胎肾脏组织中HBcAg阳性分别为6/12(50%)及10/18(55.6%)。两者之间比较无显着性差别(P>0.05)。结论死胎肾脏组织中HBcAg阳性率与孕妇HBV标志物无关,肾脏组织可能存在HBV复制。     

膜性肾病伴乙肝病毒感染的病理学改变

目的探讨肾穿刺确诊膜性肾病(membranous nephropathy,MN)合并HBV感染的病理特点及肾组织HBV抗原检测的意义。方法回顾性分析147例MN患者的临床和肾脏病理资料,根据HBV血清学结果分为A组(HBV抗原阳性)、B组(仅HBV抗体阳性)、C组(HBV阴性的原发MN),比较其肾脏病理、临床表现及实验室检测结果。肾组织HBsAg/HBcAg检测采用EnVision方法。结果 A组肾小球基膜假双轨、肾小球节段硬化、节段内皮细胞肿胀增生、内皮下和系膜区电子致密物及IgM、C4、C1q沉积的比率显著高于C组。A组的发病年龄较低,肝功能异常和低补体血症发生率明显高于B、C两组(P0.05)。A组肾组织HBsAg和(或)HBcAg阳性患者在病理及临床表现方面与阴性患者比较无明显差异(P0.05)。结论不同于原发性MN,伴HBV感染的MN常伴肾小球节段硬化、内皮细胞肿胀增生、假双轨和多种免疫球蛋白及补体多部位沉积。免疫组化未发现肾组织HBsAg和(或)HBcAg沉积不能完全排除MN与HBV感染不相关,还需结合光镜、荧光和电镜的改变综合判断。     

乙型肝炎病毒前S1抗原检测的临床意义

探讨乙型肝炎病毒前s1抗原（Pre-S1）检测在乙型肝炎病毒诊断中的临床意义。采用酶联免疫吸附试验（ELISA）和荧光定量聚合酶链反应技术（fluorescenee quantitative PCR,FQ-PCR）对650份HBV-M不同阳性模式及40份HBV—M全阴性模式血清标本进行乙型肝炎病毒Pre-S1、乙肝五项和HBV—DNA检测,并对三种检测结果进行统计学分析。在650份HBV—M不同阳性模式标本中,在119份大三阳标本中Pre—S1阳性检出率92．4％,HBV-DNA阳性检出率100％,在186份小三阳标本中Pre—SI阳性检出率42．5％,HBV—DNA阳性检出率63．4％,在21例HBsAg（＋）和HBcAb（＋）阳性组中Pres1阳性检出率47．6％,HBV．DNA阳性检出率66．7％;在297例HBsAb（＋）标本中Pre—S1阳性检出率0．4％,HBV-DNA阳性检出率0％,在268例HBV—DNA阳性的标本中Pre-S1阳性检出率79．3％。在40份HBV—M全阴模式中Pre-S1阳性检出率0％,HBV-DNA阳性检出率0％。Pre—S1在大三阳、小三阳及HBV-DNA阳性组阳性检出率明显高于阴性组（P〈0．01）,Pre—S1检测可补充和完善乙肝“两对半”检测的不足,尤其对HBeAg阴性或变异的HBV感染者能更好的反映病毒的复制状态和传染性。     

194例慢性乙型肝炎病理及其与血清HBV DNA、HBeAg、ALT关系的研究

目的探讨慢性乙型肝炎病理及其与血清HBV DNA、HBeAg、ALT关系。方法对194例慢乙肝患者进行肝组织病理、HBV免疫组化检查,并检测肝功能、血清HBVM和HBV DNA。结果血清HBeAg阳性组的肝组织G2、G3~4、S2、S3~4发生率与阴性组比较差异有统计学意义,肝组织S0组与S1~4组比较差异有统计学意义,肝组织G0~1组与G2~4组、HBcAg阳性组与阴性组的HBV DNA含量比较差异亦有统计学意义,肝组织HBsAg表达为" "者与" ~ "者血清HBV DNA含量比较差异无统计学意义,肝组织达S1或(和)G2以上者血清ALT水平分别为:<40U/L组占28.57%,40~80U/L组占53.33%,81~400U/L占80.15%,>400U/L组占77.88%。结论血清HBV DNA与肝组织HBcAg表达有一致性,与肝内HBsAg无关,HBV DNA含量低可能是肝组织炎症活动度和纤维化程度高,ASC和轻度肝损害者应争取肝活检,以及时判断肝组织病理程度和治疗时机。     

肾小球肾炎肾组织中HBV感染的标志

为了解肾组织中ＨＢＶＤＮＡ与ＨＢＶ抗原存在的关系，探讨肾组织中ＨＢＶ抗原的来源及其与病理变化的关系，我们检测２４６例肾炎肾组织中三种ＨＢＶ抗原。发现除肾小球沉积外，肾小管常有阳性表达。肾小管ＨＢｃＡｇ阳性率达２１．５４％，高于肾小球（１０．９８％）。有１８例肾组织经Ｓｏｕｔｈｅｒｎ印迹杂交检测ＨＢＶＤＮＡ，１５例阳性者中１４例肾组织ＨＢＶ抗原同时阳性，１２例血清感染标志亦阳性。结果提示某些肾炎可能与肾组织感染ＨＢＶ相关，肾组织中出现的ＨＢＶ抗原抗体免疫复合物除来自血循环外，有肾源性──原位形成的可能。     

慢性乙型肝炎血清及肝组织病毒学标志与病理损伤的关系

目的探讨慢性乙型肝炎(CHB)患者血清及肝组织病毒学标志与肝组织病理损伤的关系。方法对647例CHB患者血清病毒学标志HBsAg、HBsAb、HBeAg、HBeAb、HBcAb、HBVDNA及其中418例肝细胞病毒学标志HBsAg、HBcAg的表达与肝组织病理损伤进行对比分析。结果CHB患者血清及肝组织病毒学标志与肝组织病理损伤密切相关。结论血清HBsAg、HBeAb、HBcAb阳性,HBVDNA阴性的患者肝组织炎症及纤维化程度较轻;HBVDNA与肝组织炎症分级及纤维化分期无明显相关;肝细胞HBsAg、HBcAg均阴性表达的肝组织炎症及纤维化程度较重。     

IgA nephropathy associated with chronic hepatitis B virus infection in adults: the pathogenetic role of HBsAG

Five adult cases of IgA nephropathy associated with chronic hepatitis B virus infection were studied. Serum HBsAg and anti-HBc were present in five patients and HBeAg in four patients. Glomerular changes were typical of primary IgA nephropathy in four patients, and a mixed picture of IgA and membranous nephropathy was demonstrated in one patient. Immunofluorescence microscopy using polyclonal and monoclonal antibodies against HBsAg, HBcAg, and HBeAg revealed mesangial deposits of HBsAg in renal biopsies from four patients. One renal biopsy showed only mesangial and capillary HBcAg by polyclonal antiserum, and virus-like particles were demonstrated in the intramembranous electron-dense deposits on ultrastructural examination. Mesangial HBeAg was not detected in the renal biopsies from these patients with IgA nephropathy. As for the single patient with a mixed picture of IgA and membranous nephropathy, granular deposits of HBeAg with a distribution similar to IgG were detected in the glomerular capillary walls in addition to the mesangial deposition of HBsAg. These findings suggest that HBsAg rather than HBeAg may play a role of the pathogenesis in some of the adult patients with IgA nephropathy associated with chronic hepatitis B virus infection.     

乙型肝炎病毒前S1蛋白检测的临床意义

目的探讨乙肝前S1抗原和“乙肝六项”、HBV-DNA的关系并结合ALT的变化确定前S1抗原的临床意义和诊断价值。方法采用ELISA方法检测前S1和乙肝六项,荧光定量PCR法检测HBV-DNA,采用紫外连续监测法检测谷丙转氨酶(ALT)。结果“大三阳”标本中前S1阳性率为85.2%,HBV-DNA阳性率为90.3%;“小三阳”中前S1阳性率为46.4%,HBV-DNA阳性率为31.6%;HBSAg+、HBCAg+、e系统为阴性的前S1阳性率为26.9%,HBV-DNA的阳性率为23.0%。急性乙肝跟踪观察前S1、HBV-DNA、ALT时发现前S1能够较早反应乙肝恢复情况。ALT恢复越快前S1抗原转阴越早,且先于DNA阴转。结论前S1蛋白能够较好地反映乙肝病毒的复制情况,对病情的预后和疗效判断有较好的临床应用价值。     

Comparison of polyclonal and monoclonal antibodies in determination of glomerular deposits of hepatitis B virus antigens in hepatitis B virus-associated glomerulonephritides

The nature of hepatitis B virus (HBV) antigens in HBV-associated glomerulonephritides was investigated in 7 hepatitis B surface antigen (HBsAg) carriers with membranous nephropathy, 16 HBsAg carriers with mesangial IgA nephropathy, and 1 HBsAg carrier with a mixed picture of membranous and IgA nephropathies. Consecutive frozen sections of renal biopsy specimens were stained with polyclonal and monoclonal antibodies against HBV antigens. Glomerular capillary deposits of HBeAg and HBcAg were detected in 66% and 57% of renal biopsies from HBsAg carriers with membranous nephropathy by monoclonal and polyclonal antibodies, respectively. The discrepancy in the immunofluorescence findings resulted from the cross-reactivity of the polyclonal anti-HBcAg antiserum because it contains both anti-HBcAg and anti-HBeAg activities. Mesangial deposits of HBsAg were detected in 40% and 21% of renal biopsies from HBsAg carriers with mesangial IgA nephropathy by polyclonal and monoclonal antibodies, respectively. The authors' study confirms that HBeAg is the predominant HBV antigen deposited in HBV-associated membranous nephropathy, and glomerular HBsAg deposits are detected in some HBsAg carrier with mesangial IgA nephropathy. Careful testing and evaluation of each antibody are necessary to prevent misinterpretation.     

Peroxidase-anti-peroxidase detection of hepatitis B surface and core antigen in liver biopsy specimens from patients with chronic type B hepatitis

Liver biopsy specimens from 58 American patients with chronic type B hepatitis were investigated for the presence and distribution of the hepatitis B core (HBcAg) and surface (HBsAg) antigens by peroxidase-anti-peroxidase techniques. HBsAg was detected in 43 (77%) and HBcAg in 52 (90%) patients. HBcAg was present in 50 of 51 (98%) patients with hepatitis B e antigen (HBeAg) but in only two of seven (29%) of patients with antibody to HBeAg (anti-HBe). There was no correlation between severity of hepatitis or height of aminotransferase activities and the amount of HBsAg or HBcAg in hepatocytes but there was a positive correlation between amount of HBcAg and height of HBV-DNA and DNA polymerase activity in serum. Follow-up liver biopsies, taken 1 to 3 yr later, were available from 39 patients. HBcAg remained detectable in 25 of 26 patients with persistence of HBeAg but disappeared in 12 patients who had lost HBeAg. In nine patients, HBcAg was cytoplasmic as well as nuclear in distribution. Seven of these patients had an intense lobular hepatitis with marked elevations in aminotransferase activities. These findings indicate that the amount of HBcAg in liver correlates with the amount of serum hepatitis B virus as quantified by serum levels of DNA polymerase and HBV-DNA. The amount of nuclear HBcAg does not correlate with the severity of the liver disease, but the presence of cytoplasmic HBcAg usually reflects an active and severe ongoing hepatitis.     

早孕妇女绒毛细胞HBV感染与母婴传播关系的研究

目的探讨在携带乙型肝炎病毒的早孕孕妇中,绒毛细胞感染乙型肝炎病毒的发生情况,以及与母婴传播的关系。方法选择自愿在我院门诊行人工流产的孕妇50例,孕妇血清乙型肝炎病毒携带者为实验组（20例）,孕妇血清乙型肝炎感染标志物均阴性为对照组（30例）。分别用ELISA法检测外周血血清HBV表面标志物、荧光定量PCR法检测HBV—DNA,用免疫组织化学链霉亲和素-生物素过氧化物酶复合物（SABC）法检测孕妇的绒毛细胞。结果实验组HBV—DNA阳性16例,阴性4例。HbsAg和HbcAg均阳性5例,HbsAg和HbeAg均阳性4例,绒毛细胞出现HBV的阳性染色;对照组HBV—DNA均阴性,HbsAg和HbcAg均阴性,绒毛细胞未出现HBV的阳性染色。结论在早孕人工流产术的乙型肝炎病毒携带孕妇中,乙型肝炎病毒可感染绒毛细胞。     

HBV serum and renal biopsy markers are associated with the clinicopathological characteristics of HBV-associated nephropathy

Background: Accumulated evidence has shown that hepatitis B virus infection is associated with numerous types of nephropathy but it remains to clarify the different role of HBV markers, either in serum or deposit in kidney, in the pathogenesis of HBV-associated nephropathy. In this study, we investigated the relationship between HBV markers and HBV-associated nephropathy by using multi-linear regression in Chinese patients with HBV-associated membranous nephropathy (MN). Methods: A total of 196 cases of HBV-associated MN, which were diagnosed based on renal biopsy, were collected during the period of January 2000 to December 2009 from our hospital. Serum and renal biopsy HBV markers included HBsAg, anti-HBs, HBeAg, anti-HBe, and anti-HBC. HBV-associated nephropathy was characterized by a panel of clinical manifestations and pathological parameters, which included proteinuria, hematuria, serum creatinine, hypertension, and renal damage in glomeruli, tubules, interstitium, and blood vessels. Multilinear regression was used to analyze the relationship between the HBV markers in serum and renal biopsy and the clinicopathological characteristics of HBV-associated nephropathy. Results: After analysis of the clinical and pathological data in 196 cases of HBV-associated membranous nephropathy, this study revealed that glomerular lesion was marginally associated with serum HBsAg (P = 0.0528), Anti-HBs (P = 0.0978), but significantly associated with the presence of IgA (P = 0.0242), IgG (P < 0.0001) and C3 (P = 0.0064) in renal biopsy. There was no significant association between glomerular lesion and HBV markers in kidney. The presence of crescent and renal tube impairment was not related to HBV markers. The renal fibrosis was significantly related to gender (P = 0.023), age (P = 0.0211), HBsAg (P = 0.0001) and HBcAg (P = 0.0083) and C3 (P = 0.0299) in renal biopsy. Notably, the renal blood vessel impairment was significantly related to systolic Blood Pressure (SBP) (P < 0.0001), diastolic blood pressure (DBP) (P = 0.0002), serum HBsAg (P = 0.0428), serum HBeAg (P = 0.0766), FRA (P = 0.0002), and HBsAg (P = 0.0241) and HBcAg (P = 0.0599) in renal tissues. Also, the renal interstitial infiltration was related to patient age (P = 0.015, SBP (P < 0.0001), DBP (P = 0.0001), C3 (P = 0.0028), FRA (P = 0.0165), HBsAg (P = 0.0016) and HBcAg (P = 0.0203) in kidney biopsy. These results suggest that the major pathological changes in kidneys in HBV patients are related to one or more HBV markers, such as HBsAg, HBeAg, or anti-HBs antibody. Besides, most of the pathological changes in kidneys are related to C3 and FRA in kidney tissues. The clinical markers of nephropathy, such as proteinuria, hematuria and creatine serum levels, were also evaluated for their relationship with HBV markers in serum and kidney tissues. We found proteinuria was marginally related to HBV DNA (P = 0.0537), significantly related to IgA (0.0223). Hematouria was significantly related to IgA (P = 0.0434), IgG (P < 0.0001), and C1q (P = 0.0282). The serum creatine level was related to patient gender (P = 0.0077), SBP (P < 0.0001), DBP (0.0049), IgG (P-0.0006), and C3 (P = 0.0113). These clinical manifestations were not related to HBV markers in either serum or kidney. These results indicate that some of clinical manifestations of nephropathy are related to HBV markers, but the relationship is limited.