肿瘤坏死因子-α-238基因多态性及血清学水平与乙型肝炎病毒慢性感染结果的关系

目的 探讨肿瘤坏死因子（TNF-α）基因多态性与乙型肝炎病毒慢性感染结果之间的关系以及乙型肝炎病毒慢性感染患者血清中TNF-α水平在慢性乙肝发展中的临床意义。方法 运用聚合酶链反应-限制性片段长度多态性分析的方法检测TNF-α基因启动子区-238位点单个核苷酸多态性在不同临床类型的慢性HBV感染者及健康对照者中的分布频率;应用ELISA方法检测血清TNF-α浓度水平。结果 TNF-α-238位点G/A和G/G基因型频率以及A等位基因频率分布在实验组和健康对照组的差异无统计学意义。慢性乙型肝炎组、肝硬化组、乙型肝炎肝衰竭组和健康对照组比较,血清TNF-α均有不同程度升高,TNF-α（ng/L）取对数值后经方差分析,差异有统计学意义（P〈0.01）;组间两两比较,差异均有统计学意义（P〈0.01）,血清TNF-α水平乙型肝炎肝衰竭组〉肝硬化组〉慢性乙型肝炎组〉健康对照组。结论 TNF-α-238位点基因型与乙型肝炎易感性无明显相关性。TNF-α与乙型肝炎类型、肝损伤程度有密切关系。     

慢性HBV感染者外周血T细胞亚群与病毒载量、HBeAg的相关性分析

目的：分析慢性HBV感染者外周血T细胞亚群与病毒载量、HBeAg的相关性。方法以40例慢性乙型肝炎（ CHB）、35例慢性重型乙型肝炎（ CSHB）、30例肝硬化（ LC）及32例正常对照者为研究对象,采用流式细胞仪和荧光定量PCR法分别检测各组外周血T细胞亚群、HBV DNA载量。结果 CHB组仅CD^4＋亚群明显降低,与对照组比较,差异有统计学意义（ P ＜0．05）;CSHB及LC组CD^3＋、CD^4＋亚群、CD^4＋/CD^8＋比值呈逐渐降低趋势,CD^8＋亚群呈逐渐增高趋势,与对照组比较差异有统计学意义（ P ＜0．05）。 HBV DNA载量与CD^4＋亚群、CD^4＋/CD^8＋比值呈负相关（ r ＝-0．638,-0．778, P ＜0．05）,与CD^8＋亚群呈正相关（ r ＝0．647, P ＜0．05）,与CD^3＋无关。与HBeAg阳性患者比较,HBeAg阴性患者外周血CD^＋3＋、CD^4＋和CD^8＋亚群显著降低（ P ＜0．05）。结论慢性HBV感染者随着疾病加重细胞免疫功能进行性降低,与病毒复制水平密切相关,这是导致疾病慢性化的原因之一。外周血T细胞亚群变化趋势可为判断疾病转归及预后提供可靠指标。     

Interleukin-23 mediates the pathogenesis of LPS/GalN-induced liver injury in mice

BackgroundInterleukin-23 (IL-23) is required for T helper 17 (Th17) cell responses and IL-17 production in hepatitis B virus infection. A previous study showed that the IL-23/IL-17 axis aggravates immune injury in patients with chronic hepatitis B virus infection. However, the role of IL-23 in acute liver injury remains unclear.ObjectiveThe purpose of this study was to determine the role of the inflammatory cytokine IL-23 in lipopolysaccharide/d-galactosamine (LPS/GalN)-induced acute liver injury in mice.MethodsSerum IL-23 from patients with chronic hepatitis B virus (CHB), acute-on-chronic liver failure (ACLF) and healthy individuals who served as healthy controls (HCs) was measured by ELISA. An IL-23p19 neutralizing antibody or an IL-23p40 neutralizing antibody was administered intravenously at the time of challenge with LPS (10 μg/kg) and GalN (400 mg/kg) in C57BL/6 mice. Hepatic pathology and the expression of Th17-related cytokines, including IL-17 and TNF-α; neutrophil chemoattractants, including Cxcl1, Cxcl2, Cxcl9, and Cxcl10; and the stabilization factor Csf3 were assessed in liver tissue.ResultsSerum IL-23 was significantly upregulated in ACLF patients compared with CHB patients and HCs (P < 0.05 for both). Serum IL-23 was significantly upregulated in the non-survival group compared with the survival group of ACLF patients, which was consistent with LPS/GalN-induced acute hepatic injury in mice (P < 0.05 for both). Moreover, after treatment, serum IL-23 was downregulated in the survival group of ACLF patients (P < 0.001). Compared with LPS/GalN mice, mice treated with either an IL-23p19 neutralizing antibody or an IL-23p40 neutralizing antibody showed less severe liver tissue histopathology and significant reductions in the expression of Th17-related inflammatory cytokine, including IL-17 and TNF-α; neutrophil chemoattractants, including Cxcl1, Cxcl2, Cxcl9, and Cxcl10; and stabilization factors Csf3 within the liver tissue compared with LPS/GalN mice (P < 0.05 for all).ConclusionHigh serum IL-23 was associated with mortality in ACLF patients and LPS/GalN-induced acute liver injury in mice. IL-23 neutralizing antibodies attenuated liver injury by reducing the expression of Th17-related inflammatory cytokines, neutrophil chemoattractants and stabilization factors within the liver tissue, which indicated that IL-23 likely functions upstream of Th17-related cytokine and chemokine expression to recruit inflammatory cells into the liver.     

Increased IL-10/IL-17 ratio is aggravated along with the prognosis of patients with chronic lymphocytic leukemia

ObjectiveThis study is to investigate the association between the Treg/Th17 cells and prognosis of chronic lymphocytic leukemia (CLL).MethodsTotally 50 CLL patients and 20 Health controls were included in this study. Regulatory T (Treg) cells and the cell subset secreting IL-17 (Th17) in peripheral blood were detected with flow cytometry. Serum levels of IL-10 and IL-17 were determined with ELISA, and expression of Foxp3 and RORγt was assessed with quantitative real-time PCR.ResultsTreg and Th17 cell proportions in peripheral blood in the CLL patients were significantly higher than control. Serum levels of IL-10 and IL-17, and expression of Foxp3 and RORγt, were significantly increased in the CLL patients. Ratios of Treg/Th17 and IL-10/IL-17 were significantly elevated in the CLL patients. Compared with those before treatment, Treg/Th17 and IL-10/IL-17 ratios were declined in the CLL patients in remission. Compared with the non-remission group, Treg cells were significantly decreased, while Th17 cells were significantly increased, resulting in decreased Treg/Th17 ratio, in the remission group. Moreover, the serum IL-10 level was significantly decreased, while the serum IL-17 level was significantly increased, resulting in declined IL-10/IL-17 ratio, in the remission group. Correlation analysis showed that, Treg and Th17 cell counts were significantly associated with CD38 and ZAP-70 expression in the CLL patients. Moreover, the IL-10/IL-17 ratio was also significantly associated with CLL prognostic factors.ConclusionAltered Treg/Th17 and IL-10/IL-17 ratios in CLL would be aggravated along with the disease progression, which might be used as indicators for the disease prognosis.     

慢性乙型重型肝炎患者外周血调节性T细胞表达

目的通过观察外周血CD4^＋CD25^＋调节性T细胞（Treg）水平,探讨其在慢性乙型重型肝炎患者中的作用。方法流式细胞仪技术检测30例慢性乙型重型肝炎患者（CSHB）、30例慢性乙型肝炎患者（CHB）和15例健康志愿者（正常对照组）PBMCs中Treg的比例。结果慢性乙型重型肝炎患者外周血Treg细胞的平均百分率为（2．45±0．23）％,较慢性乙型肝炎患者的（9．48±0．48）％有极其显著差异（P〈0．01）,较健康对照组（5．65±0．61）％则有显著差异（P〈0．05）。结论Treg在慢性乙型重型肝炎的病情进展及病毒抑制清除等方面起着重要的作用。     

慢性乙型肝炎患者血CD4+CD25+调节性T细胞检测的临床意义

目的 探讨慢性乙型肝炎患者外周血CD4+CD25+调节性T细胞(CD4+ CD25+ Treg)检测的临床意义。方法 慢性HBV感染者48例（慢性HBV携带者10例为慢性HBV携带者组、慢性乙型肝炎18例慢性乙型肝炎组和乙肝肝硬化为乙肝肝硬化组20例）,以流式细胞仪检测血CD4+CD25+Treg细胞频率,实时荧光定量聚合酶链反应检测血HBV DNA载量。结果 血CD4+CD25+Treg细胞百分率：慢性HBV携带者组为(6.72±2.60)％、慢性乙型肝炎组为(8.56±3.12)％、乙肝肝硬化组为(11.59±4.34)％,组间差异有统计学意义（均P＜0.05）。血HBV DNA载量：慢性HBV携带者组为1.7 ×106拷贝/ml、慢性乙型肝炎组为4.3 ×105拷贝/ml、乙肝肝硬化组为6.8 ×104拷贝/ml,组间差异有统计学意义(P＜0.01);慢性肝病患者血CD4+CD25+调节性T细胞百分率与HBV DNA滴度正相关。结论 慢性肝病患者血Treg细胞频率与HBVDNA滴度正相关,提示Treg细胞在慢性乙型肝炎致病机制中起重要作用。     

Toll-like receptor 7 and 9 expression in peripheral blood mononuclear cells from patients with chronic hepatitis B and related hepatocellular carcinoma

AIM: The aim of the present study was to investigate the expression of Toll-like receptors (TLR) 7 and 9 in peripheral blood mononuclear cells (PBMC) of patients with chronic hepatitis B virus (HBV) infection and related hepatocellular carcinoma. METHODS: The study group was comprised of 52 patients: 41 with chronic hepatitis B and 11 healthy controls. The protein and mRNA levels of TLR7 and TLR9 were evaluated using real-time PCR, Western blot analysis, and flow cytometry. We also detected the serum viral load of HBV in the patients and analyzed the correlation between HBV-DNA copies and the TLR expression. RESULTS: Our results demonstrated a lower TLR7 expression in all HBV infection groups compared to the controls. We found that HBV infection led to a decreased expression of TLR9 mRNA, but an increased expression of the TLR9 protein compared to the healthy group. The TLR protein levels are related to serum HBV-DNA (P<0.01). CONCLUSION: There are downregulations of TLR7 expression and TLR9 mRNA in PBMC of HBV-infected patients, but an increased TLR9 expression at the protein level.     

HBV感染者血清白介素(12,2,10)的水平及临床意义

目的检测HBV不同感染状态时血清IL-12、IL-2和IL-10水平,并探讨其临床意义。方法选择HBV不同感染状态,包括无症状携带者、无症状携带者重叠急性自限性肝炎、HBeAg阳性慢性乙型肝炎、HBeAg阴性慢性乙型肝炎和急性乙型肝炎患者等作为研究对象,以健康献血员作对照,采用双抗体夹心ELISA法检测血清IL-12、IL-2和IL-10水平并进行比较。结果无症状携带者血清IL-10水平明显高于健康对照,部分无症状携带者血清IL-12和IL-2高水平;重叠感染HAV、HEV感染IL-2、IL-10和IL-12水平急性期和恢复期相比无明显差异;慢性乙型肝炎患者发病急性期血清IL-12水平低下,血清IL-2和IL-10水平均较健康对照明显升高;急性乙型肝炎患者血清IL-12和IL-2水平与ALT水平并非完全平行关系,血清IL-10水平与ALT水平呈完全平行关系。结论血清IL-12和IL-2水平可考虑作为无症状携带者抗病毒治疗指征,慢性乙型肝炎活动与血清IL-12水平低下有关,血清IL-2水平升高与肝炎活动有关,其升高主要参与促进HBeAg/HBeAb血清转换。肝炎活动与血清IL-10水平无明显相关性。     

饮酒与乙肝病毒感染对肝脏损伤的协同作用及机制的研究进展

饮酒和乙肝病毒感染是造成肝脏损伤的两大病因。流行病学调查显示,在慢性肝病患者中,饮酒及HBV感染两种损伤因素多并存。饮酒与HBV感染促进肝脏损伤的发生和进展,促进肝细胞癌的发生。饮酒可促进体内HBV的病毒复制和基因突变,而HBV感染亦可增加肝细胞对乙醇损伤的易感性。这些研究为临床肝病患者的保健和治疗提供了依据。本文从临床表现、临床研究和实验机制研究等方面综述了饮酒与HBV感染对肝脏损伤的协同作用及其机制。     

46例慢性HBV感染者临床与肝组织病理及免疫组化相关性分析

目的 探讨HBV感染者的肝脏病理和免疫组化与部分临床生化指标的关系.方法 采用一秒钟经皮肝脏穿刺活检法取得肝组织,进行病理形态学观察以及免疫组化法检测组织中HBsAg的表达,同步检测部分血清生化指标进行统计学分析.结果 慢乙肝轻、中度的临床与病理诊断符合率分别为72.7%和40%,肝硬化的符合率为18.1%.慢乙肝患者肝组织中HBsAg表达与炎症分级G无相关性,与TB升高程度呈负相关.肝组织的炎症分级G与ALT、纤维化分期S呈正相关.纤维化分期S与ALT、TB无相关性.结论 慢乙肝和肝硬化的临床诊断和病理诊断一致性较低.慢性HBV感染者病毒清除可以造成肝细胞的损伤,但并不是主要原因.     

慢性乙型肝炎血清HBV DNA载量与肝组织病理的关系

目的探讨HBeAg阳性和HBeAg阴性慢性乙型肝炎血清HBV DNA载量与肝组织炎症活动度、纤维化程度之间的关系。方法分别对103例HBeAg阳性和81例HBeAg阴性慢性乙型肝炎患者肝脏穿刺活检,进行肝组织炎症活动度分级和纤维化程度分期,同期检测血清HBV DNA载量,并分析两者之间的关系。结果 103例HBeAg阳性慢性乙型肝炎患者血清HBV DNA载量与肝组织炎症活动度、纤维化程度均无明显相关性(r=0.125,P>0.05;r=0.164,P>0.05)。81例HBeAg阴性慢性乙型肝炎患者血清HBV DNA载量与肝组织炎症活动度、纤维化程度均呈正相关(r=0.326,P<0.01;r=0.267,P<0.05)。结论 HBeAg阴性慢性乙型肝炎患者血清HBV DNA载量能反映肝组织炎症活动度和纤维化程度,可作为抗病毒药物选择和疗效观察的指标。     

慢性HBV感染患者HBV DNA前C区变异的临床意义

目的:探讨慢性HBV感染患者HBVDNA前C区变异的临床意义。方法:采用ELISA测定99例HBV感染患者血清肝炎病毒标志物,并采用RT-PCR检测HBVDNA前C区变异。结果:99例HBV感染患者中,乙型肝炎病毒携带26例,慢性乙型病毒性肝炎61例,慢性重型病毒性肝炎(乙型)11例和急性乙型病毒性肝炎1例。乙型肝炎病毒携带,慢性乙型病毒性肝炎和慢性重型病毒性肝炎中,变异株组HBVDNA前C区变异发生率与混合株组、野生株组相比,差异均无显著性(P>0.05)。与混合株和野生株两组相比,HBVDNA变异株组肝功能变化差异均无显著性(P>0.05)。结论:HBVDNA前C区变异可发生于HBV感染的不同临床状态。HBVDNA前C区变异可能与肝炎程度和病情进展无明显关系。     

Lamivudine. A review of its therapeutic potential in chronic hepatitis B.

Lamivudine is a deoxycytidine analogue that is active against hepatitis B virus (HBV). In patients with chronic hepatitis B, lamivudine profoundly suppresses HBV replication. Clinically significant improvements in liver histology and biochemical parameters were obtained with lamivudine in double-blind, randomised, trials in hepatitis B e antigen (HBeAg)-positive patients with chronic hepatitis B and compensated liver disease. After 52 weeks of treatment, relative to placebo (< or = 25%), significantly more Chinese (56%) or Western patients (52%) treated with lamivudine 100 mg/day had reductions of > or = 2 or more points in Knodell necro-inflammatory scores. Moreover, significantly fewer lamivudine 100 mg/day than placebo recipients had progressive fibrosis in liver biopsies (< or = 5 vs > or = 15%) and fewer lamivudine- than placebo-treated patients progressed to cirrhosis (1.8 vs 7.1%). More lamivudine 100 mg/day than placebo recipients acquired antibodies to HBeAg after 52 weeks (16 vs 4% in Chinese patients and 17 vs 6% in Western patients). ALT levels normalised in significantly more lamivudine than placebo recipients enrolled in these trials. In HBeAg-negative, HBV DNA positive patients with compensated liver disease enrolled in a double-blind, randomised study, HBV DNA levels were suppressed to below the limit of detection (< 2.5 pg/ml) and ALT levels normalised in 63% and 6% of patients treated with lamivudine 100 mg/day or placebo for 24 weeks. Clinically significant improvements in liver histology were obtained in 60% of patients treated with lamivudine for 52 weeks in this study. Lamivudine 100 mg/day for 52 weeks produced similar or significantly greater improvements in liver histology and ALT levels than 24 weeks' treatment with lamivudine plus interferon-alpha. In liver transplant candidates with chronic hepatitis B and end-stage liver disease, lamivudine 100 mg/day alone, or in combination with hepatitis B immune globulin, generally suppressed HBV replication and appeared to protect the grafted liver from reinfection. Lamivudine 100 mg/day suppressed viral replication and improved liver histology in liver transplant recipients with recurrent or de novo chronic hepatitis B. Lamivudine 300 or 600 mg/day reduced HBV replication in HIV-positive patients. The incidence of adverse events in patients with chronic hepatitis B and compensated liver disease treated with lamivudine 100 mg/day or placebo for 52 to 68 weeks was similar. 3.1- to 10-fold increases in ALT over baseline occurred in 13% of patients during treatment with lamivudine 100 mg/day or placebo for 52 weeks. Post-treatment ALT elevations were more common in lamivudine than placebo recipients; however, these generally resolved spontaneously; < or = 1.5% of lamivudine- or placebo-treated patients experienced hepatic decompensation. CONCLUSION: Lamivudine inhibits HBV replication, reduces hepatic necro-inflammatory activity and the progression of fibrosis in patients with chronic hepatitis B, ongoing viral replication and compensated liver disease including HBeAg-negative patients. The drug also suppresses viral replication in liver transplant recipients and HIV-positive patients. Thus, lamivudine is potentially useful in a wide range of patients with chronic hepatitis B and ongoing viral replication.     

Circulating CTLA-4 levels and CTLA4 polymorphisms associate with disease condition and progression and hepatocellular carcinoma patients' survival in chronic hepatitis B virus infection

BackgroundCTLA-4 is involved in the immune dysfunction of hepatitis B virus (HBV) infection and hepatocellular carcinoma (HCC). This study analyzed the association of circulating CTLA-4 levels and CTLA4 polymorphisms with disease condition and progression in chronic HBV infection.MethodsSerum CTLA-4 levels and CTLA4 rs231775 and rs5742909 polymorphisms were determined in patients with various HBV-related diseases [53 asymptomatic HBV carrier status (ASC), 147 chronic hepatitis, 130 cirrhosis and 102 HCC] and nearly a 10-year follow-up.ResultsSerum CTLA-4 levels were stepwisely increased from ASC, chronic hepatitis, cirrhosis to HCC and independently associated with HCC (OR 2.628, P < 0.001). HCC patients had lower frequencies of rs231775 genotype GA, genotype AA and allele A than ASC, chronic hepatitis and cirrhosis patients. Rs231775 genotype GG was independently associated with HCC (OR 2.324, P = 0.010) and higher CTLA-4 levels in patients with HBV infection. In the follow-up, higher baseline CTLA-4 levels and CTLA4 rs231775 genotype GG significantly associated with disease progression from chronic hepatitis to cirrhosis (OR 2.561, P = 0.011 and OR 2.799, P = 0.015, respectively) or from cirrhosis to HCC (OR 2.673, P = 0.008 and OR 2.097, P = 0.023, respectively) and with a shorter overall survival in HCC patients (HR 0.317, P = 0.018 and HR 0.682, P = 0.026, respectively). Rs5742909 had no significant association with CTLA-4 levels and disease progression.ConclusionCTLA-4 levels and CTLA4 rs231775 polymorphism associate with the disease condition and progression and HCC development in chronic HBV infection and their determination may be used for monitoring disease progression and predicting patient prognosis.     

免疫球蛋白 A肾病病人血清白细胞介素 -17、白细胞介素 -18表达变化与乙型肝炎病毒感染的相关性

目的探讨免疫球蛋白 A(IgA)肾病病人血清白细胞介素 -17(IL-17)、白细胞介素 -18(IL-18)表达变化与乙型肝炎病毒(HBV)感染的相关性。方法选取 2016年 3月至 2019年 3月南阳市中心医院 224例 IgA肾病病人,其中乙型肝炎病毒感染 67例作为感染组,未感染病人 157例作为未感染组。另选取无肾脏疾病的单纯乙型肝炎病毒感染病人 67例作为对照组。对比三组不同乙肝病毒脱氧核糖核酸( HBV-DNA)载量病人、乙肝表面抗原( HBeAg)阴性及阳性病人血清 IL-17、IL-18水平,分析血清 IL-17、IL-18对 IgA肾病病人 HBV感染的诊断价值及与 HBV-DNA载量、肝功能［谷草转氨酶( AST)、谷丙转氨酶( ALT)］相关性。结果三组血清 IL-17、IL-18水平对比差异有统计学意义( P<0.05),且未感染组 IL-17(12.68±2.13)ng/L、IL-18(360.37±     

慢性乙肝病毒携带者的细胞免疫功能变化及其与血清HBV DNA的关系

目的　探讨慢性乙肝病毒携带者的细胞免疫功能变化与血清 HBVDNA关系。方法　应用流式细胞仪直接免疫荧光法检测 5 0例慢性乙肝病毒携带者外周血 T淋巴细胞亚群百分率 ,并以 2 0例正常健康人为对照组进行比较。结果　乙肝病毒携带者外周血 CD3+ 细胞百分率与正常对照组比较 ,无统计学差异 (P>0 .0 5 )。 Th细胞百分率较正常对照组明显降低 (P<0 .0 5 ) ;Tc细胞百分率则明显升高 (P<0 .0 1 ) ;Th/ Tc明显降低 (P<0 .0 1 )。HBVD-NA (+)组与 HBVDNA (- )组相比 ,CD3+ 细胞百分率无统计学差异 (P>0 .0 5 ) ,Th细胞百分率有降低趋势 ,Tc细胞百分率有升高趋势 ,而 Th/ Tc明显下降 (P<0 .0 1 )。结论　 HBV感染可导致乙肝病毒携带者细胞免疫功能改变 ,HBVDNA复制增加可进一步导致乙肝病毒携带者细胞免疫功能紊乱