目的探讨血清丛生蛋白(Clusterin)及趋化因子受体5(CCR5)在慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)患者中的表达以及两者联合检测对ACLF患者预后的预测价值。方法选取2018年1月~2020年12月唐山市传染病医院收治的84例ACL...目的探讨血清丛生蛋白(Clusterin)及趋化因子受体5(CCR5)在慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)患者中的表达以及两者联合检测对ACLF患者预后的预测价值。方法选取2018年1月~2020年12月唐山市传染病医院收治的84例ACLF患者作为观察组,同期选择在该院体检的80例健康体检者作为对照组。采用酶联免疫吸附试验(ELISA)法测定患者血清中Clusterin和CCR5水平;根据患者预后28天情况,将其分为存活组(n=48)和死亡组(n=36)。采用受试者工作特征曲线(receiver operating characteristic curve,ROC)分析血清Clusterin与CCR5联合检测对ACLF患者预后的预测价值;Spearman相关性分析血清Clusterin和CCR5水平与终末期肝病模型(model for end-stage liver disease,MELD)评分、慢性肝衰竭-序贯器官衰竭评分(chronic liver failure-sequential organ failure assessment,CLIF-SOFA)的相关性;采用多因素Logistic回归分析ACLF患者预后的影响因素。结果与对照组相比,观察组Clusterin(87.37±9.99μg/ml vs 104.85±15.14μg/ml)及CCR5(11.55±2.86μg/ml vs 15.68±3.01μg/ml)水平降低,差异具有统计学意义(t=8.767,9.010,均P<0.05)。与存活组相比,死亡组Clusterin(77.40±9.26μg/ml vs94.85±10.54μg/ml)及CCR5(8.58±1.98μg/ml vs 13.78±3.52μg/ml)水平降低,差异具有统计学意义(t=7.904,7.962,均P<0.05)。ROC曲线显示,血清Clusterin与CCR5联合预测的曲线下面积(area under curve,AUC)(0.927)最大,其敏感度和特异度分别为88.90%和83.30%。经Spearman相关性分析Clusterin与MELD,CLIF-SOFA评分呈负相关(r=-0.524,-0.457,均P<0.05),CCR5与MELD,CLIF-SOFA评分呈负相关(r=-0.611,-0.358,均P<0.05)。多因素Logistic回归分析显示,血清Clusterin,CCR5及IL-6为ACLF患者预后不良的影响因素(均P<0.05)。结论Clusterin及CCR5在ACLF患者血清中表达下调,且联合检测二者在预测ACLF患者短期预后方面具有良好的参考价值。展开更多
Objective: To study the function of the first extracellular membrane loop of the NH2-terminal of chemokine receptor 5(CCRS), and produce its specific antibody F(ab’ )2. Methods: Some amino acid sequences of β chemok...Objective: To study the function of the first extracellular membrane loop of the NH2-terminal of chemokine receptor 5(CCRS), and produce its specific antibody F(ab’ )2. Methods: Some amino acid sequences of β chemokine superfamily receptors were aligned by computer and the least homologous domain of the extracellular loops was located. In this paper, the first extracellular domain (114 nucleotides) was defined and amplified by PCR, and a expression vector of the recombinant GST fusion protein,pGEXIN/NR5, was constructed. After the inserted sequence was confirmed correct, the transformation and expression of fusion protein was performed in E. coli and the fusion protein was purified. Then 2 Newzealand rabbits were immunized. The anti-CCR5 NH2-terminal antibody F(ab’ )2 was made by protein A affinity chromatography, pepsin digestion and sepharose-12 column chromatography. Results: Reduced and unreduced SDS-PAGE and FAX analysis demonstrated that this F(ab’ )2 had a high specificity to combine with CCR5. Conclusion: A simple and quick method to prepare specific antibody F(ab’ )2 of certain functional domain is showed in this paper, by which we can get an improtant experiment material for studying gene expression, and it will provide a good idea and a technique to study other high similar superfamily members.展开更多
文摘目的探讨血清丛生蛋白(Clusterin)及趋化因子受体5(CCR5)在慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)患者中的表达以及两者联合检测对ACLF患者预后的预测价值。方法选取2018年1月~2020年12月唐山市传染病医院收治的84例ACLF患者作为观察组,同期选择在该院体检的80例健康体检者作为对照组。采用酶联免疫吸附试验(ELISA)法测定患者血清中Clusterin和CCR5水平;根据患者预后28天情况,将其分为存活组(n=48)和死亡组(n=36)。采用受试者工作特征曲线(receiver operating characteristic curve,ROC)分析血清Clusterin与CCR5联合检测对ACLF患者预后的预测价值;Spearman相关性分析血清Clusterin和CCR5水平与终末期肝病模型(model for end-stage liver disease,MELD)评分、慢性肝衰竭-序贯器官衰竭评分(chronic liver failure-sequential organ failure assessment,CLIF-SOFA)的相关性;采用多因素Logistic回归分析ACLF患者预后的影响因素。结果与对照组相比,观察组Clusterin(87.37±9.99μg/ml vs 104.85±15.14μg/ml)及CCR5(11.55±2.86μg/ml vs 15.68±3.01μg/ml)水平降低,差异具有统计学意义(t=8.767,9.010,均P<0.05)。与存活组相比,死亡组Clusterin(77.40±9.26μg/ml vs94.85±10.54μg/ml)及CCR5(8.58±1.98μg/ml vs 13.78±3.52μg/ml)水平降低,差异具有统计学意义(t=7.904,7.962,均P<0.05)。ROC曲线显示,血清Clusterin与CCR5联合预测的曲线下面积(area under curve,AUC)(0.927)最大,其敏感度和特异度分别为88.90%和83.30%。经Spearman相关性分析Clusterin与MELD,CLIF-SOFA评分呈负相关(r=-0.524,-0.457,均P<0.05),CCR5与MELD,CLIF-SOFA评分呈负相关(r=-0.611,-0.358,均P<0.05)。多因素Logistic回归分析显示,血清Clusterin,CCR5及IL-6为ACLF患者预后不良的影响因素(均P<0.05)。结论Clusterin及CCR5在ACLF患者血清中表达下调,且联合检测二者在预测ACLF患者短期预后方面具有良好的参考价值。
文摘Objective: To study the function of the first extracellular membrane loop of the NH2-terminal of chemokine receptor 5(CCRS), and produce its specific antibody F(ab’ )2. Methods: Some amino acid sequences of β chemokine superfamily receptors were aligned by computer and the least homologous domain of the extracellular loops was located. In this paper, the first extracellular domain (114 nucleotides) was defined and amplified by PCR, and a expression vector of the recombinant GST fusion protein,pGEXIN/NR5, was constructed. After the inserted sequence was confirmed correct, the transformation and expression of fusion protein was performed in E. coli and the fusion protein was purified. Then 2 Newzealand rabbits were immunized. The anti-CCR5 NH2-terminal antibody F(ab’ )2 was made by protein A affinity chromatography, pepsin digestion and sepharose-12 column chromatography. Results: Reduced and unreduced SDS-PAGE and FAX analysis demonstrated that this F(ab’ )2 had a high specificity to combine with CCR5. Conclusion: A simple and quick method to prepare specific antibody F(ab’ )2 of certain functional domain is showed in this paper, by which we can get an improtant experiment material for studying gene expression, and it will provide a good idea and a technique to study other high similar superfamily members.
