原发性胆汁性肝硬化患者肝硬化期外周血淋巴细胞计数及T细胞亚群检测与分析

目的探讨原发性胆汁性肝硬化(PBC)患者肝硬化期外周血淋巴细胞总数及T细胞亚群的特点并分析影响因素及临床意义。方法对86例肝硬化期PBC患者的临床资料、实验诊断数据进行回顾性分析,比较40例代偿期和46例失代偿期PBC患者肝功、免疫学指标和外周血淋巴细胞亚群特点,对可能的影响因素进行相关性分析。结果失代偿期PBC患者年龄、血清总胆红素(TBil)水平、血清总IgA水平、Mayo评分高于代偿期患者(P0.05),而ALT、ALB、外周血淋巴细胞绝对数(LYMPH)、淋巴细胞百分率(LYMPH%)、T淋巴细胞绝对数(CD3+)、T辅助细胞绝对数(CD3+CD4+)和T抑制细胞绝对数(CD3+CD8+)均低于代偿期患者(P0.05);LYMPH、LYMPH%、CD3+、CD3+CD4+和CD3+CD8+结果中,失代偿期减低者频率均高于代偿期(45.7%vs.10%,34.8%vs.7.5%,58.7%vs.17.5%,45.7%vs.5%,60.9%vs.27.5%,P0.05),失代偿期增高者频率均低于代偿期(54.3%vs.90%,6.5%vs.27.5%,0 vs.20%,2.2%vs.22.5%,2.2%vs.10%,P0.05)。在可能的影响因素中,年龄、Mayo评分、上消化道出血、脾大或脾切除、腹水等肝硬化失代偿表现对结果影响呈负相关;而ALT、ALB对结果影响呈正相关。结论随着肝硬化程度的加深,PBC患者外周血淋巴细胞总数及T细胞亚群数量降低;从淋巴细胞定量角度分析,随着疾病的进展,PBC患者免疫水平下降。     

抗中性粒细胞胞浆抗体对自身免疫性肝炎的检测意义

目的探讨抗中性粒细胞胞浆抗体（ANCA）对自身免疫性肝炎的检测意义。方法选择210例肝功能异常患者,其中自身免疫性肝炎（AIH）41例,原发性胆汁性肝硬化（PBC）19例,原发性硬化性胆管炎（PSC）3例,其他肝功能异常患者147例。应用间接免疫荧光法和免疫印迹法检测ANCA及其他相关自身抗体,并对患者临床资料进行分析比较。结果本组患者中,ANCA阳性16例,占肝功能异常患者的7.6%。16例ANCA阳性患者中AIH11例,PSC3例,慢性乙型肝炎1例,药物性肝损伤1例。41例AIH中,11例ANCA阳性（11/41,26.8%）,同时伴有ANA阳性,核型以均质型为主（8/11,72.7%）;30例ANCA阴性者中,25例ANA阳性,核型多样化,均质型仅占20.0%（6/30）。结论本研究中AIH中的ANCA阳性率仅为26.8%,以高滴度为主;ANCA阳性AIH患者ANA均阳性,其核型以均质型为主,且未观察到特殊的临床特征。     

抗乙醇脱氢酶抗体ELISA法的建立及其对自身免疫性肝炎诊断的价值

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for the detection of serum alcohol dehydrogenase (ADH) antibody and evaluate its role in its diagnosis of autoimmune hepatitis( AIH ). Methods The reactivity between yeast ADH and human anti-ADH serum antibody was tested by Western blot analysis. ELISA was established using yeast ADH. The method was applied in serums of 67 AIH patients,94 primary biliary cirrhosis(PBC) patients, 199 chronic hepatitis B (CHB) patients, 132 chronic hepatitis(CHC) patients, 24 alcohol hepatitis disease(ALD) patients, 99 connective tissue disease(CTD) patients and 31 healthy individuals. The positive rate of ADH antibody in the patients and healthy individuals was measured. The χ2 test was used to compare the positive rates. Results The ELISA method for detecting human anti-ADH serum antibody was established successfully and the optimum reaction conditions were defined. Western blot showed that yeast ADH has cross reactivity with human anti-ADH antibody. The positive rate of anti-ADH antibody in the AIH group [59. 7% ,40/67 ] was higher than that in the normal control group(0,χ2 = 31. 271 ,P <0. 05), PBC group (6. 4% ,χ2 =54. 492,P <0. 05), CHB group( 14. 1% ,χ2 =54. 848,P <0. 05) ,CHC group(21.2% ,χ2 = 29.269,P<0.05), ALl) group ( 25. 0% ,χ2 =8.512,P <0.05)and CTD group ( 43. 4% ,χ2 =4.229, P <0. 05). Conclusions Compared with the PBC, CHB, CHC, ALD and CTD group, the anti-ADH antibody positive rate in the serums of AIH was significantly increased. The antibody may be helpful to the diagnosis of autoimmune hepatitis.     

一种HBV DNA 定量检测试剂的准确性分析

目的 评价一种新型HBV DNA 定量检测试剂的准确性。方法 将第3 代HBV DNA WHO 国 际标准品稀释为6 种不同梯度浓度样本,8.5×105、8.5×104、8.5×103、8.5×102、85 及42.5 IU/ml,用待评价 试剂careHBV PCR Assay V3.0 检测,并对实测值和理论值进行相关性分析;以Cobas Taqman HBV V2.0 为参 比试剂,用careHBV PCR Assay V3.0 检测93 份HBV 感染者标本和30 份健康者标本HBV DNA 含量,并对2 种试剂定量检测结果进行相关性和一致性分析;采用巢式PCR 扩增漏检标本的HBV S 区,直接测序法测定 其基因型。结果 careHBV PCR Assay V3.0 检测WHO 国际标准品的实测值和理论值具有线性相关;2 种定 量试剂检测93 份HBV 感染者标本结果均阳性共68 份,检测结果呈线性相关,差异无统计学意义（P >0.05）, 但careHBV PCR Assay V3.0 检测值相对偏低;careHBV PCR Assay V3.0 检测有14 份标本漏检,其中13 份标 本Cobas Taqman HBV V2.0 检测结果位于30 ～ 2 000 IU/ml 间;对1 份漏检标本的基因型检测结果为C 型。 结论 careHBV PCR Assay V3.0 与Cobas Taqman HBV V2.0 试剂检测结果有较好的相关性,但对基因型C 型 的标本可能存在漏检现象。     

抗乙醇脱氢酶抗体ELISA法的建立及其对自身免疫性肝炎诊断的价值

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for the detection of serum alcohol dehydrogenase (ADH) antibody and evaluate its role in its diagnosis of autoimmune hepatitis( AIH ). Methods The reactivity between yeast ADH and human anti-ADH serum antibody was tested by Western blot analysis. ELISA was established using yeast ADH. The method was applied in serums of 67 AIH patients,94 primary biliary cirrhosis(PBC) patients, 199 chronic hepatitis B (CHB) patients, 132 chronic hepatitis(CHC) patients, 24 alcohol hepatitis disease(ALD) patients, 99 connective tissue disease(CTD) patients and 31 healthy individuals. The positive rate of ADH antibody in the patients and healthy individuals was measured. The χ2 test was used to compare the positive rates. Results The ELISA method for detecting human anti-ADH serum antibody was established successfully and the optimum reaction conditions were defined. Western blot showed that yeast ADH has cross reactivity with human anti-ADH antibody. The positive rate of anti-ADH antibody in the AIH group [59. 7% ,40/67 ] was higher than that in the normal control group(0,χ2 = 31. 271 ,P <0. 05), PBC group (6. 4% ,χ2 =54. 492,P <0. 05), CHB group( 14. 1% ,χ2 =54. 848,P <0. 05) ,CHC group(21.2% ,χ2 = 29.269,P<0.05), ALl) group ( 25. 0% ,χ2 =8.512,P <0.05)and CTD group ( 43. 4% ,χ2 =4.229, P <0. 05). Conclusions Compared with the PBC, CHB, CHC, ALD and CTD group, the anti-ADH antibody positive rate in the serums of AIH was significantly increased. The antibody may be helpful to the diagnosis of autoimmune hepatitis.     

原发性胆汁性肝硬化患者外周血单个核细胞Toll样受体表达与调节性T细胞相关性分析

目的 探讨原发性胆汁性肝硬化(PBC)患者外周血单个核细胞(PBMC)Toll样受体2、4、9(TLR2,TLR4,TLR9)表达及PBMC中CD4+ CD25+调节性T细胞(Tregs)特点和影响因素.方法 采用流式细胞术检测PBC患者(52例)和健康对照者(22例)外周血PBMC中TLR2、4、9阳性细胞及Tregs比例,比较其在PBC患者和健康对照者中的差别并分析TLR与Tregs相关性.结果 PBC患者Tregs比例低于健康对照者[(1.53+1.33)vs(4.42±1.43),P＜0.0001],TLR4阳性细胞比例高于健康对照者[(36.95±3.53)vs(32.84±8.06),P=0.003];PBC患者Tregs比例与TLR9阳性细胞比例呈负相关(R2 =0.115,P=0.016),健康对照者Tregs比例与TLR2,4和TLR2,4,9联合表达阳性细胞比例呈负相关(R2=0.326,P=0.007;R2 =0.226,P=0.034).结论 PBC患者外周血PBMC中Tregs比例降低,TLR4表达升高,可能受肝硬化失代偿期腹腔感染的影响.     

抗乙醇脱氢酶抗体ELISA法的建立及其对自身免疫性肝炎诊断的价值

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for the detection of serum alcohol dehydrogenase (ADH) antibody and evaluate its role in its diagnosis of autoimmune hepatitis( AIH ). Methods The reactivity between yeast ADH and human anti-ADH serum antibody was tested by Western blot analysis. ELISA was established using yeast ADH. The method was applied in serums of 67 AIH patients,94 primary biliary cirrhosis(PBC) patients, 199 chronic hepatitis B (CHB) patients, 132 chronic hepatitis(CHC) patients, 24 alcohol hepatitis disease(ALD) patients, 99 connective tissue disease(CTD) patients and 31 healthy individuals. The positive rate of ADH antibody in the patients and healthy individuals was measured. The χ2 test was used to compare the positive rates. Results The ELISA method for detecting human anti-ADH serum antibody was established successfully and the optimum reaction conditions were defined. Western blot showed that yeast ADH has cross reactivity with human anti-ADH antibody. The positive rate of anti-ADH antibody in the AIH group [59. 7% ,40/67 ] was higher than that in the normal control group(0,χ2 = 31. 271 ,P <0. 05), PBC group (6. 4% ,χ2 =54. 492,P <0. 05), CHB group( 14. 1% ,χ2 =54. 848,P <0. 05) ,CHC group(21.2% ,χ2 = 29.269,P<0.05), ALl) group ( 25. 0% ,χ2 =8.512,P <0.05)and CTD group ( 43. 4% ,χ2 =4.229, P <0. 05). Conclusions Compared with the PBC, CHB, CHC, ALD and CTD group, the anti-ADH antibody positive rate in the serums of AIH was significantly increased. The antibody may be helpful to the diagnosis of autoimmune hepatitis.     

抗乙醇脱氢酶抗体ELISA法的建立及其对自身免疫性肝炎诊断的价值

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for the detection of serum alcohol dehydrogenase (ADH) antibody and evaluate its role in its diagnosis of autoimmune hepatitis( AIH ). Methods The reactivity between yeast ADH and human anti-ADH serum antibody was tested by Western blot analysis. ELISA was established using yeast ADH. The method was applied in serums of 67 AIH patients,94 primary biliary cirrhosis(PBC) patients, 199 chronic hepatitis B (CHB) patients, 132 chronic hepatitis(CHC) patients, 24 alcohol hepatitis disease(ALD) patients, 99 connective tissue disease(CTD) patients and 31 healthy individuals. The positive rate of ADH antibody in the patients and healthy individuals was measured. The χ2 test was used to compare the positive rates. Results The ELISA method for detecting human anti-ADH serum antibody was established successfully and the optimum reaction conditions were defined. Western blot showed that yeast ADH has cross reactivity with human anti-ADH antibody. The positive rate of anti-ADH antibody in the AIH group [59. 7% ,40/67 ] was higher than that in the normal control group(0,χ2 = 31. 271 ,P <0. 05), PBC group (6. 4% ,χ2 =54. 492,P <0. 05), CHB group( 14. 1% ,χ2 =54. 848,P <0. 05) ,CHC group(21.2% ,χ2 = 29.269,P<0.05), ALl) group ( 25. 0% ,χ2 =8.512,P <0.05)and CTD group ( 43. 4% ,χ2 =4.229, P <0. 05). Conclusions Compared with the PBC, CHB, CHC, ALD and CTD group, the anti-ADH antibody positive rate in the serums of AIH was significantly increased. The antibody may be helpful to the diagnosis of autoimmune hepatitis.     

抗乙醇脱氢酶抗体ELISA法的建立及其对自身免疫性肝炎诊断的价值

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for the detection of serum alcohol dehydrogenase (ADH) antibody and evaluate its role in its diagnosis of autoimmune hepatitis( AIH ). Methods The reactivity between yeast ADH and human anti-ADH serum antibody was tested by Western blot analysis. ELISA was established using yeast ADH. The method was applied in serums of 67 AIH patients,94 primary biliary cirrhosis(PBC) patients, 199 chronic hepatitis B (CHB) patients, 132 chronic hepatitis(CHC) patients, 24 alcohol hepatitis disease(ALD) patients, 99 connective tissue disease(CTD) patients and 31 healthy individuals. The positive rate of ADH antibody in the patients and healthy individuals was measured. The χ2 test was used to compare the positive rates. Results The ELISA method for detecting human anti-ADH serum antibody was established successfully and the optimum reaction conditions were defined. Western blot showed that yeast ADH has cross reactivity with human anti-ADH antibody. The positive rate of anti-ADH antibody in the AIH group [59. 7% ,40/67 ] was higher than that in the normal control group(0,χ2 = 31. 271 ,P <0. 05), PBC group (6. 4% ,χ2 =54. 492,P <0. 05), CHB group( 14. 1% ,χ2 =54. 848,P <0. 05) ,CHC group(21.2% ,χ2 = 29.269,P<0.05), ALl) group ( 25. 0% ,χ2 =8.512,P <0.05)and CTD group ( 43. 4% ,χ2 =4.229, P <0. 05). Conclusions Compared with the PBC, CHB, CHC, ALD and CTD group, the anti-ADH antibody positive rate in the serums of AIH was significantly increased. The antibody may be helpful to the diagnosis of autoimmune hepatitis.     

抗乙醇脱氢酶抗体ELISA法的建立及其对自身免疫性肝炎诊断的价值

目的 建立血清抗乙醇脱氢酶(alcohol dehydrogenase,ADH)抗体ELISA法,并评价抗.ADH在诊断自身免疫性肝炎(AIH)中的价值.方法 用免疫印迹试验对酵母ADH与人血清抗.ADH之间的反应性进行验证.用酵母ADH建立检测血清抗-ADH的ELISA法.以67例AIH、94例原发性胆汁性肝硬化(PBC)、199例慢性乙型肝炎(CHB)、132例慢性丙型肝炎(CHC)、24例酒精性肝病(ALD)和99例结缔组织病(CTD)患者及31名健康对照者为研究对象,对其血清中的抗-ADH进行检测并统计其阳性率,阳性率的比较用χ2检验.结果 建立了一种检测人血清抗-ADH的ELISA法,并确定了最佳反应条件;免疫印迹试验证实酵母ADH与人血清抗-ADH有良好反应性.AIH患者血清抗-ADH阳性率为59.7%(40/67),高于健康对照组(0,χ2=31.271,P<0.05)、PBC组(6.4%,χ2=54.492,P<0.05)、CHB组(14.1%,χ2=54.848,P<0.05)、CHC组(21.2%,χ2=29.269,P<0.05)、ALD组(25.0%,χ2=8.512,P<0.05)和CTD组(43.4%,χ2=4.229.P<0.05).结论 AIH患者血清中抗-ADH阳性率高于其他肝病和某些自身免疫性疾病患者,可能对AIH有一定辅助诊断价值.