黑素瘤缺乏因子2在乙型肝炎病毒相关性肾小球肾炎发病机制中的作用

目的 初步探讨黑素瘤缺乏因子2(AIM2)激活与表达在HBV相关性肾小球肾炎(HBV-GN)发病机制中的作用.方法 54例HBV-GN患者为研究组,25例慢性肾小球肾炎患者为阴性对照组,6例慢性乙型肝炎患者为阳性对照组.免疫组织化学法分别测定研究组、阴性对照组患者肾组织及阳性对照组患者肝组织中AIM2、Caspase-1、IL-1β的表达.组间比较采用卡方检验,相关性分析采用Spearman相关分析.结果 54例HBV-GN患者肾组织中AIM2的阳性表达率为81.4％,25例慢性肾小球肾炎组为4.0％,两组间差异有统计学意义(x2=38.746,P＜0.01).AIM2在6例慢性乙型肝炎患者肝组织中均有表达.HBV-GN组AIM2的表达强度与Caspase-1呈正相关(rs=0.444,P＜0.01),IL-1β的表达强度与Caspase-1也呈正相关(rs=0.515,P＜0.01).HBV-GN患者中血HBV DNA≥1×105拷贝/mL者AIM2的表达强度高于HBV DNA＜1×105拷贝/mL者(x2=6.097,P＜0.05).结论 AIM2的激活与HBV-GN有关,可能通过Caspase-1激活,释放炎性因子IL-1β.     

乙型肝炎病毒相关性肾炎中Toll样受体4沉积特点及其意义

目的 观察HBV相关性肾炎(HBV-GN)患者肾组织Toll样受体4(TLR4)的表达及分布情况,探讨TLR4与HBV-GN肾组织病变及临床表现的关系.方法 采用免疫组化法检测经肾活检确诊的48例HBV-GN患者及154例非HBV-GN患者的肾组织标本,观察肾组织TLR4的分布特征,明确TLR4与HBsAg的分布关系,结合病史资料,统计并分析其与病理类型,肾小球、肾小管病变程度,肾间质炎性细胞浸润程度及血清HBV等临床指标之间的关系.结果 TLR4以棕红色颗粒状物主要沉积在HBV-GN组患者的肾小管及间质中,且与HBsAg的分布基本一致.HBV-GN组TLR4阳性率及阳性积分均高于非HBV-GN组(P＜0.05).在系膜增生性肾小球肾炎组、局灶节段性肾小球硬化症组TLR4阳性积分略高,但差异无统计学意义(P＞0.05);肾小管病变程度与TLR4的表达显著相关(r =0.748,P＜0.001),且随肾小管萎缩、炎性细胞浸润及间质纤维化的程度加重而升高(r分别为0.577、0.684、0.569,P值均＜0.001),但与肾小球的病变程度无显著相关性(r=0.293,P=0.053).回归分析发现,TLR4表达随血压的升高、GFR的下降、高敏C反应蛋白的增加及24h尿蛋白定量的增多而增加(R2分别为0.869、0.784、0.979、0.615),其余临床指标均无统计学意义.结论 TLR4在HBV-GN肾组织中有异常表达,主要分布于肾小管上皮细胞及间质组织,其分布与HBsAg分布基本一致,表达水平与炎性细胞浸润、肾间质病变及肾功能变化密切相关.故推测,HBV-GN肾组织中的HBV可促进TLR4的异常表达,后者可能参与了HBV-GN的肾脏组织病变的进展.     

乙型肝炎病毒相关性肾炎患者病毒基因型及变异的临床分析

目的检测乙型肝炎病毒相关性肾炎(HBV-GN)患者的HBV基因型,探讨HBV S区、P区变异与HBV-GN病情变化的相关性。方法对10例确诊HBV-GN患者取血,抽提全基因组DNA,DNA测序仪检测HBV P区、S区序列,进行病毒基因分型,并查找突变位点。结果 10例HBV-GN患者检出HBV基因型C型9例(占90%),B型1例(占10%);出现YVDD变异1例,同义突变1例,各占10%。结论本组HBV基因型以C型为主,发生变异少,未发现HBV变异与HBV-GN病情变化的相关性。     

乙型肝炎病毒相关性肾炎的临床病理特征及治疗

目的探讨乙型肝炎病毒相关性肾炎(HBV-GN)的病理特征、治疗方法及其疗效。方法选取2012年2月至2015年2月我院收治的HBV-GN患者61例,根据HBV DNA水平将其分为两组(A组为HBV DNA水平103拷贝数/mL;B组为HBV DNA水平≤103拷贝数/mL),分别对其用药[A组-强的松(0.5 mg/kg)+血管紧张素转化酶抑制剂(ACEI)+拉米夫定(100 mg),B组-强的松(0.5 mg/kg)+ACEI],比较两组患者治疗前后生化指标的改变,同时对其病理特征进行分析。结果①患者的病理表现主要为膜性肾病(62.3%)、膜增殖性肾炎(31.1%)、系膜增生性肾炎(6.6%);②A组治疗前后差异具有统计学意义的生化指标有HBV DNA、血清白蛋白以及24 h尿蛋白定量(P均(.05);B组有HBV DNA、血清白蛋白、24 h尿蛋白定量以及血肌酐(P均0.05)。结论 HBV-GN的临床症状多样、病理类型复杂,应加强肾组织活检中HBV DNA的检查,以提高检出率;治疗方案应根据其具体病程或身体状态,应用ACEI等优化整体疗效。     

拉米夫定联合雷公藤多苷治疗乙型肝炎病毒相关性肾炎的研究

目的：探讨拉米夫定联合雷公藤多苷治疗乙型肝炎病毒相关性肾炎的疗效及安全性。方法我科住院并行肾穿刺活检术确诊的36例HBV-GN患者,所有患者口服拉米夫定100 mg/d、雷公藤多苷0．5 mg · kg -1· d-1,治疗12个月。观察24小时尿蛋白定量、血清白蛋白、血肌酐、ALT、HBV DNA ,并行肾组织病理及 HBV抗原检测。结果拉米夫定联合雷公藤多苷治疗 HBV-GN ,35例患者13例获得 CR （37．1％）,16例获得 PR （45．7％）。总有效率为82．8％,治疗效果随治疗时间延长而提高。治疗后蛋白尿、ALT明显下降,血清白蛋白明显升高（P＜0．05）,血肌酐未见明显变化（P＞0．05）。治疗后11例HBeAg消失,6例出现抗HBe ,HBV DNA明显下降。结论拉米夫定联合雷公藤多苷是治疗HBV-GN安全有效的方法,不良反应少,值得临床借鉴。     

IgA肾病患者肾组织中HBV DNA表达状况的研究

为了探讨乙型肝炎病毒(HBV)感染在IgA肾病发病中的作用，采用免疫组化技术检测12例IgA肾病患者肾组织中HBV抗原，同时应用Southern印迹杂交和原位分子杂交技术检测肾组织中HBV DNA的存在状态及定位情况。结果表明12例IgA肾病患者血清HBV感染标志至少一项阳性；肾组织中HBV抗原均阳性，其中HBcAg在肾小管和肾小球中阳性分别为8例和5例，HBsAg阳性分别为4例和6例；10例中有8例用Southern印迹杂交技术证实存在整合型HBV DNA；原位杂交技术证实12例患者肾组织HBV DNA均阳性，定位于肾小管上皮细胞核内，其中9例在肾小球系膜细胞核、上皮细胞核和基质中同时阳性。提示除了HBV抗原抗体复合物所致体液免疫损伤机制外，亦应考虑肾组织感染HBV导致的细胞免疫机制参与了IgA肾病的发病。     

肾组织伴有或无乙型肝炎病毒抗原沉积的老年患者临床和病理对比分析

目的通过对肾组织伴有或无乙型肝炎病毒(HBV)抗原沉积的老年患者的临床和病理资料进行对比分析,进一步探讨老年乙肝相关性肾炎的临床和病理特点。方法对联勤保障部队第九〇〇医院年龄≥60岁的33例乙肝相关性肾炎(HBV-GN)和27例乙肝合并肾炎(HBV-CG)患者的临床、病理资料进行回顾性分析及统计学比较。结果(1)两组均以男性[26例(78.8%)和21例(77.8%)]多见,临床表现均以肾病综合征[18例(54.5%)和15例(55.6%)]为主,其中HBV-GN组合并高血压者明显多于HBV-CG组;(2)两组均以膜性肾病(MN)为主要病理类型,其中HBV-GN组病理类型为膜增生性肾小球肾炎(MPGN)者明显多于HBV-CG组;(3)HBV-GN组在"大三阳"及高病毒载量者所占比例上均明显高于HBV-CG组;(4)HBV-GN组肾组织可观察到多种免疫复合物沉积的现象,即"满堂亮";(5)HBV-GN组病理改变不同于HBV-CG组之处:具有不典型MN的表现;各病理类型肾组织中偶可观察到新月体形成;局灶节段肾小球硬化(FSGS)中可有基底膜增厚的表现;(6)HBV-GN组在肾小管及间质病变的发生率及病变程度上均明显高于HBV-CG组。结论尽管老年HBV-GN与HBV-CG在临床表现及检验指标上有诸多相似之处,但HBV-GN者在血清病毒学及病理改变上仍与HBV-CG者存在较大的区别。因此对于该类老年患者,应重视血清HBV复制及肾脏病理改变情况,进而为治疗提供临床依据。     

激光微分离检测肾组织中病毒DNA在乙型肝炎相关性肾炎分子诊断中的应用

目的：建立应用激光微分离检测肾活检组织切片中乙型肝炎病毒(HBV)DNA的方法，提高乙肝相关性肾炎临床诊断的水平。方法：对11例外周病毒血清学HBsAg：HBcAb和HBeAg阳性、排除其他继发性。肾小球疾病，肾活检病理类型为膜性。肾病或膜增生性肾病的患者，运用激光微分离系统分离。肾活检组织切片中的肾小球和。肾小管，以PCR法检测血清HBV—DNA和用免疫组织化学方法检测。肾组织HBsAg、HBcAg和HBeAg。结果：6例肾小球HBV—DNA阳性，5例阴性。与肾组织病毒抗原的检测结果对照发现，1例肾组织中未检测到病毒抗原或DNA，2例HBV-DNA阳性而病毒抗原阴性，4例同时检测到病毒抗原和DNA，其余4例仅检测到病毒抗原的沉积。肾组织：HBV—DNA检出率(54．5％)低于病毒抗原总的检出率(72．7％)。结论：激光微分离结合PCR检测肾组织乙肝病毒DNA，取材精准，快捷高效，重复性好，为临床上乙肝相关性肾炎的诊断提供了一个新的有效的途径。     

HBV相关性肾炎肾组织AIM2水平及其与炎性因子和肾脏炎症程度的相关性分析

目的分析乙型肝炎病毒相关性肾炎(HBV-GN)患者肾组织黑色素瘤缺乏因子2(AIM2)水平及其与炎性因子和肾脏炎症程度的相关性。方法收集本院62例HBV-GN患者(HBV-GN组)和46例慢性肾小球肾炎(CGN)患者(CGN组)的临床资料,行肾脏穿刺获取肾组织标本,制备石蜡进行包埋切片处理。采用免疫组织化学法检测两组患者肾组织中的AIM2水平和白细胞介素-1β(IL-1β)、半胱氨酸天冬氨酸蛋白酶-1(caspase-1)等炎性因子的水平,进而采用Spearman相关性分析探究AIM2水平与HBV-GN患者炎性因子和肾脏炎症程度的关系。结果在HBV-GN患者的肾组织中,AIM2主要分布于肾小球内皮细胞和系膜细胞中。HBV-GN组AIM2的阳性表达率为79.03%,较CGN组的32.61%明显升高(P0.05)。HBV-GN患者肾组织中AIM2水平与IL-1β、caspase-1均存在正相关关系(r=0.58、0.68,均P0.01)。结论 HBV-GN患者肾组织AIM2表达水平明显升高,且与IL-1β、caspase-1等多种炎性因子水平和肾脏炎症程度存在密切关系。     

乙型肝炎病毒感染肾炎患者肾组织病毒抗原和复制中间体的检测及其意义

目的：观察乙型肝炎病毒(HBV)感染者肾组织中三种病毒抗原成分的分布特点及其与HBV感染状态和临床病理之间的联系，探讨在肾组织局部是否存在HBV的复制。方法：免疫组化法检测合并HBV感染的30例膜性肾病和12例膜增生性肾炎病例的肾活检组织切片中的HBsAg、HBcAg和HBeAg，同时检测肾小球和循环中的HBV基因组DNA及其复制中间体——闭合环状双链DNA(cccDNA)。结果：膜性肾病肾组织中病毒抗原的检出率(83．3％)显著高于膜增生性肾炎(33％)；膜性肾病肾组织检出的抗原以HBc舷和HBeAg多见，其中，血清HBeAg阳性病例肾组织HBeAg的检出率显著高于HBeAg阴性的病例。膜增生性肾炎肾组织检出的抗原主要是HBeAg。肾组织HBeAg的检出与循环中HBeAg的存在明显相关。伴血清转氨酶升高者肾组织HBV抗原的检出率较转氨酶正常者有升高的趋势。肾小球HBVDNA和cccDNA的检出均与循环中的检测结果高度一致，并以伴活动性HBV感染者检出率为高。结论：在合并HBV感染的肾炎患者中，肾组织HBV抗原的检出率在膜性肾病患者明显高于膜增生性肾炎。肾小球中检出的HBV抗原成分以HBeAg和HBcAg最多见，肾小球HBe他的检出与血清中是否存在HBeAg明显相关。合并肝功能损害者肾组织HBV抗原的检出率较肝功能正常者有增高趋势。在乙肝相关性肾炎患者的肾小球中确实能检测到HBV复制中间体的存在，它的出现与循环中HBV复制中间体检出的高度一致性，不能排除循环中HBV感染细胞在肾组织潴留对结果的影响，其意义还有待进一步阐明。     

成人与小儿肾脏组织中HBV－DNA存在状态的比较

本文分析８例成人与７例小儿肾脏疾病的肾组织中检出的ＨＢＶ－ＤＮＡ。临床诊断：７例小儿均为乙肝病毒相关性肾炎（ＨＢＶ－ＧＮ）；８例成人中３例为ＨＢＶ－ＧＮ，其余为血尿待查、狼疮性肾炎和ＩｇＡ肾病。ＨＢＶ－ＤＮＡ在肾组织的存在状态在成人均为整合型，而小儿有整合型及游离型二种。小儿肾组织中ＨＢｓＡｇ阳性表达率高于成人。作者认为此现象可能与成人肾组织中ＨＢＶ－ＤＮＡ存在状态仅有一种整合型有关。另外发现肾组织中ＨＤｃＡｇ阳性率与ＨＢＶ－ＤＮＡ呈正相关。肾组织中ＨＢｃＡｇ阳性的患者局部有Ｔ细胞的浸润，提示了肾原性抗体存在及其在ＨＢＶ－ＧＮ中激发细胞免疫参与肾脏病变可能性。     

Further study on the immunopathology of hepatitis B virus associated glomerulonephritis

By using immunohistochemical techniques the deposition of HBV associated immune complexes was studied in 845 consecutive cases of renal biopsy. In 665 cases of primary glomerulonephritis the frequencies of HBsAg, HBeAg and HBcAg detection in glomeruli were 11.9%, 8.3% and 3.2% respectively with a total HBV antigen positive frequency of 12.2%. High positive rates were found in membranous glomerulonephritis (MGN, 37.1%), mesangioproliferative GN (MPGN, 26%) and IgA nephropathy (IgA-NP, 18.9%). The detection of HBV infection markers in serum were simultaneously performed in 213 cases; 31.7% of the patients with primary GN were found to be positive. In patients with positive HBV infectious markers in the serum, deposits of HBV antigens in glomeruli were found in 49.1% of the cases. The incidence was significantly different in the serum negative group (10.6%). Meanwhile, about 68.3% of the cases with HBV antigen deposits in the kidney was found to have positive HBV markers in the serum. Also the incidence was significantly different in the group without HBV antigen deposits in the kidney (20.9%). It was again confirmed that the pathogenesis of hepatitis B virus associated glomerulonephritis (HBV-GN) was related to the deposition of HBV immune complexes in kidney tissue. It was noticed that the deposition of three different types of HBV antigens was somewhat associated with the development of specific forms of HBV GN. The diagnostic criteria of HBV-GN were discussed in detail.     

对拉米夫定治疗无应答或变异的乙型肝炎病毒相关性肾炎的治疗

目的探讨对拉米夫定治疗无应答或变异的乙型肝炎病毒相关性肾炎(HBV-GN)的治疗方法。方法 2005年1月至2009年1月南京军区福州总医院诊治的HBV-GN且对拉米夫定治疗无应答或变异的患者共9例。采用恩替卡韦(0.5 mg/d)为主方案治疗9例对拉米夫定治疗无应答或变异的HBV-GN且蛋白尿>1.5 g/d、血HBV-DNA≥1.0×108拷贝/L的患者,观察其疗效及血HBV-DNA的变化。结果 9例患者肾病综合征(NS)7例、蛋白尿伴血尿2例,其中系膜增生性肾炎3例,膜性肾病3例,膜增生性肾炎、IgA肾病及局灶节段系膜增生性肾炎各1例。拉米夫定治疗(14.1±10.3)个月(其中6例联合激素治疗),5例部分缓解(PR)、4例无效(均为NS),改用恩替卡韦治疗后,治疗12个月时7例完全缓解(CR)、1例PR、1例NR,7例检测血清HBV-DNA水平患者中5例降至正常,随访观察了19～27个月,平均(23.7±3.0)个月,至随访结束,完全缓解(CR)7例,NR 2例,均停用激素。未见副反应。结论恩替卡韦为主的治疗对对拉米夫定治疗无应答或变异的HBV-GN是安全有效的,疗程以1年半为宜。     

The genotype C could play a key role in hepatitis B virus associated nephritis among the Northwest Chinese children

BackgroundHepatitis B virus-associated glomerulonephritis (HBV-GN) is a kind of immune complex-induced glomerulonephritis. The present study was designed to determine whether Hepatitis B virus (HBV) genotype is associated with glomerulonephritis in north-west Chinese children.MethodsA total of 296 HBV-infected patients were enrolled in this study. The serum of patients was subjected to DNA extraction and the HBV genotypes were determined by PCR.ResultsThe results showed that genotype C (49%) was predominant within the subjects, compared to HBV/B (38.5%), B/C recombinant (7.4%) and none B/C (5.1%). The serum tests showed that the changes of Complement 3 (C3) and alanine amino transferase (ALT) levels in the genotype C patients were significantly greater than those in the genotype B patients. The frequency of genotype C in HBV-GN patients was higher than that in non HBV-GN patients (χ2 value = 30.239, P < 0.001). But, it was not associated with renal dysfunction. Furthermore, The genotype C was associated with high HBV-DNA load (82.9% vs 17.1%, P < 0.001), which is seen more frequently in the HBV-GN children (86.3% vs 13.3%, P = 0.004).ConclusionsThe genotype C may play a role in HBV-GN children, via favoring HBV replication.     

Chronic hepatitis B serum promotes apoptotic damage in human renal tubular cells

AIM: To investigate the effect of the serum of patients with chronic hepatitis B (CHB) on apoptosis of renal tubular epithelial cells in vitro and to study the role of hepatitis B virus (HBV) and transforming growth factor-β1 (TGF-β1) in the pathogenesis of hepatitis B virus associated glomerulonephritis (HBV-GN). METHODS: The levels of serum TGF-β1 were measured by specific enzyme linked immunosorbent assay (ELISA) and HBV DNA was tested by polymerase chain reaction (PCR) in 44 patients with CHB ,and 20 healthy persons as the control. The normal human kidney proximal tubular cell (HK-2) was cultured together with the sera of healthy persons, CHB patients with HBV-DNA nega-tive(20 cases) and HBV-DNA positive (24 cases) for up to 72 h. Apoptosis and Fas expression of the HK-2 were detected by flow cytometer. RESULTS: The apoptosis rate and Fas expression of HK-2 cells were significantly higher in HBV DNA positive serum group 19.01±5.85% and 17.58±8.35%, HBV DNA negative serum group 8.12±2.80% and 6.96±2.76% than those in control group 4.25±0.65% and 2.33±1.09%, respectively (P < 0.01). The apoptosis rate and Fas expression of HK-2 in HBV DNA positive serum group was significantly higher than those in HBV DNA negative serum (P < 0.01). Apoptosis rate of HK-2 cells in HBV DNA positive serum group was positively correlated with the level of HBV-DNA (r = 0.657). The level of serum TGF-β1 in CHB group was 163.05±91.35μg/L, significantly higher as compared with 81.40±40.75μg/L in the control group (P < 0.01). CONCLUSION: The serum of patients with chronic hepatitis B promotes apoptotic damage in human renal tubular cells by triggering a pathway of Fas up-regula-tion. HBV and TGF-β1 may play important roles in the mechanism of hepatitis B virus associated glomerulone-phritis.     

HBV感染肾小管细胞机制的研究进展

肾脏具备HBV感染肝细胞的关键因素,HBV相关肾小球肾炎(HBV-GN)的发病机制尚未完全明确,近年来HBV直接损伤肾小管的机制越来越受到重视。HBV可以通过调节细胞周期、激活NF-κB等相关信号转导通路诱导肾小管细胞凋亡,导致HBV-GN进展。目前在临床上尚无作用靶点位于肾脏治疗HBV-GN的药物。将HBV感染细胞的关键因素和HBV损伤肾小管细胞研究进展总结如下,阐述HBV直接感染肾小管细胞的可能机制,为HBV-GN的治疗提供新思路。     

Meta-analysis of combined therapy for adult hepatitis B virus-associated glomerulonephritis

AIM: To investigate the efficacy and safety of combined antiviral and immunosuppressant therapy in adult hepatitis B virus-associated glomerulonephritis (HBV-GN) patients.METHODS: A computerized literature search was carried out in the PubMed database, Embase, the Cochrane Library, Chinese BioMedical Literature on disc, Chinese Medical Current Contents, Chinese National Knowledge Infrastructure, Wanfang and VIP (Chinese Technological Journal of Database) to collect articles between June 1980 and December 2010 on therapy with immunosuppressants, e.g., glucorticosteroids, mycophenolate mofetil and leflunomide, combined with antivirals, e.g., interferon, lamivudine, entecavir and adefovir dipivoxil, in adult HBV-GN patients. The primary outcomes were remission of proteinuria, clearance of HBV e-antigen, and elevation of serum albumin. The secondary outcomes were blood levels of alanine aminotransferase, serum creatinine, and HBV-DNA titer. Meta-analysis was performed using Review Manager 5.1. Fixed or random effect models were employed to combine the results after a heterogeneity test. The effects of the combined therapy were analyzed for different doses of glucorticosteroid and different types of HBV-GN.RESULTS: Twelve clinical trials with 317 patients were included. A significantly higher incidence of HBV-GN was found in male patients (relative risk = 2.40, 95% CI: 1.98-2.93). Combined therapy reduced the proteinuria significantly with a mean difference of 4.19 (95% CI: 3.86-4.53) and increased the serum albumin concentration significantly with a mean difference of -11.95 (95% CI: -12.97-10.93) without significant alterations of liver function (mean difference: 4.62, 95% CI: -2.55-11.79) and renal function (mean difference: 10.29, 95% CI: 0.14-20.45). No significant activation of HBV-DNA replication occurred (mean difference: 0.12, 95% CI: -0.37-0.62). There was no significant difference between the high dose glucorticosteroid group and the low dose glucorticosteroid group in terms of proteinuria remission (P = 0.76) and between different pathological types of HBV-GN [membranous glomerulonephritis (MN) vs mesangial proliferative glomerulonephritis, P = 0.68; MN vs membranoproliferative glomerulonephritis, P = 0.27].CONCLUSION: Combined antiviral and immunosuppressant therapy can improve the proteinuria in HBV-GN patients without altering HBV replication or damaging liver and renal functions.     

乙型肝炎病毒相关性肾炎内皮素、补体的变化

探讨补体及内皮素 (ET)在乙型肝炎病毒相关性肾炎 (HBV -GN)中的变化。采用同位素放射免疫方法检测正常人及HBV -GN患者血、尿内皮素的水平 ;用单项免疫扩散法测定血清补体水平。HBV -GN患者血、尿ET水平均较对照组明显增高 ,而血清补体水平均较对照组降低。内皮素及补体在HBV -GN的发病机理及病情进展中可能起重要作用 ,其值高低可作为判断肾损害严重程度及预后的指标