Objectives Autoantibodies play a key role in mechanism of rheumatic heart disease(RHD) and in this research,we focus on identifying the autoantibody in Chinese RHD patients which can be a potential molecular biomarker...Objectives Autoantibodies play a key role in mechanism of rheumatic heart disease(RHD) and in this research,we focus on identifying the autoantibody in Chinese RHD patients which can be a potential molecular biomarker. Methods To construct RHD expression library,the total RNA of heart tissue of RHD patients was extracted and mRNA was isolated,purified and reverse-transcripted to long cDNA.Phage was used to recombining cDNA to be expression library.The library was immunoscreened by serum of active rheumatic fever patients,An autoantigen positive clone was immunoscreened.To identify the autoantibody,autoantibody gene was analysed by PCR,sequencing and bioinfor-matics. It also was subcloned and expressed in vitro.Western blotting was used to identify the expression protein and test cross-reaction effect with serum.Results An expression library with heart tissue of RHD patients was successfully established. The titer of the primary library was 3.3×10~6 pfu/ml, recombinant rate of it was 99%and 81%inserted segments were larger than 1 kb.A positive antoantibody gene was sreened and it is homologous to keratin 18.The recombined vector could expression objective protein in vitro which could be reacted with sera of active rheumatic fever patients and rheumatic heart disease patients,but could not be detected by sera of health persons.Conclusions It is an effective strategy to investigate autoantigens of Chinese RHD that constructing and immunoscreening heart tissues expression library. This research implied that keratin 18 was a candidate molecular biomarker of RHD.展开更多
Objective To analyze the serum keratin 18(K18)level of patients with hepatitis B virus(HBV)-related acute-on-chronic liver failure(HBV-ACLF)and its correlation with prognosis.Methods From December 2012 to October 2014...Objective To analyze the serum keratin 18(K18)level of patients with hepatitis B virus(HBV)-related acute-on-chronic liver failure(HBV-ACLF)and its correlation with prognosis.Methods From December 2012 to October 2014,120 patients who visited Fuzhou Infectious Diseases Hospital and were diagnosed with HBV-ACLF were enrolled,and 20 chronic hepatitis B(CHB)pa-展开更多
目的探讨乙型肝炎病毒相关性慢加急性肝衰竭(HBV-ACLF)患者血清角蛋白18(K-18)水平变化及其对患者预后的预测价值。方法 2015年1月～2017年1月我院收治的HBV-ACLF患者100例,另选择性别和年龄相匹配的慢性乙型肝炎(CHB)患者30例和健康人3...目的探讨乙型肝炎病毒相关性慢加急性肝衰竭(HBV-ACLF)患者血清角蛋白18(K-18)水平变化及其对患者预后的预测价值。方法 2015年1月～2017年1月我院收治的HBV-ACLF患者100例,另选择性别和年龄相匹配的慢性乙型肝炎(CHB)患者30例和健康人30例。采用酶联免疫吸附试验法检测血清K18(M30和M65)水平。随访3 m。应用受试者工作特征曲线(ROC)分析M30/M65比值和终末期肝病模型(MELD)预测患者预后的效能。结果 ACLF患者血清M30、M65水平和M30/M65比值分别为(1.0±0.1) lg U/L、(3.4±0.3) lg U/L和(0.3±0.1),与CHB患者【分别为(2.1±0.1) lg U/L、(3.3±0.2) lg U/L和(0.6±0.0)】或者健康人【分别为(2.1±0.1) lg U/L、(2.1±0.1) lg U/L和(1.0±0.2)】比,有显著性统计学差异(P<0.05);68例生存患者MELD评分为(18.2±.0),显著低于32例死亡患者的【(26.1±3.3),P<0.05】,血清M65水平为(2.9±0.2) lg U/L,显著低于死亡患者的【(4.1±0.5) lg U/L,P<0.05】,M30/M65比值为(0.4±0.1),显著高于死亡患者的【(0.2±0.1),P<0.05】;MELD评分预测ACLF患者死亡的截断点为23.21,其AUC为0.650,预测的灵敏度、特异度和正确性分别为85.5%、65.0%和72.0%,而M30/M65比值预测ACLF患者死亡的截断点为0.30,其AUC为0.875,预测的灵敏度、特异度和正确性分别为94.5%、85.0%和85.0%,显著优于MELD评分(P<0.05)。结论 K18(M30和M65)与肝脏疾病的严重程度有关。早期检测血清M30和M65水平可能有助于对HBV-ACLF患者预后的判断。展开更多
文摘Objectives Autoantibodies play a key role in mechanism of rheumatic heart disease(RHD) and in this research,we focus on identifying the autoantibody in Chinese RHD patients which can be a potential molecular biomarker. Methods To construct RHD expression library,the total RNA of heart tissue of RHD patients was extracted and mRNA was isolated,purified and reverse-transcripted to long cDNA.Phage was used to recombining cDNA to be expression library.The library was immunoscreened by serum of active rheumatic fever patients,An autoantigen positive clone was immunoscreened.To identify the autoantibody,autoantibody gene was analysed by PCR,sequencing and bioinfor-matics. It also was subcloned and expressed in vitro.Western blotting was used to identify the expression protein and test cross-reaction effect with serum.Results An expression library with heart tissue of RHD patients was successfully established. The titer of the primary library was 3.3×10~6 pfu/ml, recombinant rate of it was 99%and 81%inserted segments were larger than 1 kb.A positive antoantibody gene was sreened and it is homologous to keratin 18.The recombined vector could expression objective protein in vitro which could be reacted with sera of active rheumatic fever patients and rheumatic heart disease patients,but could not be detected by sera of health persons.Conclusions It is an effective strategy to investigate autoantigens of Chinese RHD that constructing and immunoscreening heart tissues expression library. This research implied that keratin 18 was a candidate molecular biomarker of RHD.
文摘Objective To analyze the serum keratin 18(K18)level of patients with hepatitis B virus(HBV)-related acute-on-chronic liver failure(HBV-ACLF)and its correlation with prognosis.Methods From December 2012 to October 2014,120 patients who visited Fuzhou Infectious Diseases Hospital and were diagnosed with HBV-ACLF were enrolled,and 20 chronic hepatitis B(CHB)pa-
文摘目的探讨乙型肝炎病毒相关性慢加急性肝衰竭(HBV-ACLF)患者血清角蛋白18(K-18)水平变化及其对患者预后的预测价值。方法 2015年1月～2017年1月我院收治的HBV-ACLF患者100例,另选择性别和年龄相匹配的慢性乙型肝炎(CHB)患者30例和健康人30例。采用酶联免疫吸附试验法检测血清K18(M30和M65)水平。随访3 m。应用受试者工作特征曲线(ROC)分析M30/M65比值和终末期肝病模型(MELD)预测患者预后的效能。结果 ACLF患者血清M30、M65水平和M30/M65比值分别为(1.0±0.1) lg U/L、(3.4±0.3) lg U/L和(0.3±0.1),与CHB患者【分别为(2.1±0.1) lg U/L、(3.3±0.2) lg U/L和(0.6±0.0)】或者健康人【分别为(2.1±0.1) lg U/L、(2.1±0.1) lg U/L和(1.0±0.2)】比,有显著性统计学差异(P<0.05);68例生存患者MELD评分为(18.2±.0),显著低于32例死亡患者的【(26.1±3.3),P<0.05】,血清M65水平为(2.9±0.2) lg U/L,显著低于死亡患者的【(4.1±0.5) lg U/L,P<0.05】,M30/M65比值为(0.4±0.1),显著高于死亡患者的【(0.2±0.1),P<0.05】;MELD评分预测ACLF患者死亡的截断点为23.21,其AUC为0.650,预测的灵敏度、特异度和正确性分别为85.5%、65.0%和72.0%,而M30/M65比值预测ACLF患者死亡的截断点为0.30,其AUC为0.875,预测的灵敏度、特异度和正确性分别为94.5%、85.0%和85.0%,显著优于MELD评分(P<0.05)。结论 K18(M30和M65)与肝脏疾病的严重程度有关。早期检测血清M30和M65水平可能有助于对HBV-ACLF患者预后的判断。
