目的本研究旨在探索细胞因子在结直肠癌患者与健康者之间的差异表达,及筛选出的高表达细胞因子与肿瘤标志物联合检测在结直肠癌诊疗中的作用。方法利用细胞因子微阵列分析结直肠癌患者和健康人血浆细胞因子表达谱;采用酶联免疫法(ELISA...目的本研究旨在探索细胞因子在结直肠癌患者与健康者之间的差异表达,及筛选出的高表达细胞因子与肿瘤标志物联合检测在结直肠癌诊疗中的作用。方法利用细胞因子微阵列分析结直肠癌患者和健康人血浆细胞因子表达谱;采用酶联免疫法(ELISA)检测筛选出的结直肠癌患者血浆中高表达细胞因子。用受试者工作特征(ROC)曲线评价癌胚抗原(CEA)、糖类抗原125(CA125)、CA199和经过筛选获得的细胞因子:受激活调节正常T细胞表达和分泌因子(RANTES)在鉴别结直肠癌患者和健康者的诊断性能。所有数据使用SPSS23.0和Graph Pad Prism 8.0进行统计分析,验证CEA、CA125、CA199和RANTES在结直肠癌患者和健康者之间差异表达采用Mann-Whitney U检验。通过计算Youden指数来确定RANTES水平的最佳临界值,以鉴别结直肠癌患者。用二元Logistic回归模型结合血清/血浆生物标志物的诊断效果进行拟合。所有统计检验均为双侧检验,差异有统计学意义(P<0.05)。结果与健康者相比,结直肠癌患者血浆RANTES显著高表达,差异有统计学意义(P<0.001)。RANTES的ROC曲线下面积为0.846(95%CI:0.791~0.901);RANTES和CEA联合检测AUC为0.891(95%CI:0.854~0.929),灵敏度和特异度分别达到0.854和0.803。结论RANTES对结直肠癌有诊断价值,RANTES联合CEA检测较单独检测CEA有更高的结直肠癌诊断效能。展开更多
An expected 276bp fragment of the gene precursor encoding the signal peptide and mature protein of human β chemokine RANTES was amplified by reverse transcription\|polymerase chain reaction (RT\|PCR) from RNA of PHA\...An expected 276bp fragment of the gene precursor encoding the signal peptide and mature protein of human β chemokine RANTES was amplified by reverse transcription\|polymerase chain reaction (RT\|PCR) from RNA of PHA\|activated human peripheral blood lymphocytes.This putative interested gene was inserted directly into a T\|vector and the ligation was confirmed by restriction enzyme digestion.The sequence data of the cloned fragment showed that it was almost identical with published sequences of RANTES gene,except for only one nucleotide substitution within the signal peptide region.The \%in vitro\% expressed recombinant RANTES protein was detected by the chemiluminescence enzyme\|linked immune Dot blotting assay after combining the recombinant plasmid with the \%in vitro\% SP6/T7 transcription and translation system.The successful cloning and expression of RANTES gene should shed light on future′s gene therapy of AIDS.展开更多
文摘目的本研究旨在探索细胞因子在结直肠癌患者与健康者之间的差异表达,及筛选出的高表达细胞因子与肿瘤标志物联合检测在结直肠癌诊疗中的作用。方法利用细胞因子微阵列分析结直肠癌患者和健康人血浆细胞因子表达谱;采用酶联免疫法(ELISA)检测筛选出的结直肠癌患者血浆中高表达细胞因子。用受试者工作特征(ROC)曲线评价癌胚抗原(CEA)、糖类抗原125(CA125)、CA199和经过筛选获得的细胞因子:受激活调节正常T细胞表达和分泌因子(RANTES)在鉴别结直肠癌患者和健康者的诊断性能。所有数据使用SPSS23.0和Graph Pad Prism 8.0进行统计分析,验证CEA、CA125、CA199和RANTES在结直肠癌患者和健康者之间差异表达采用Mann-Whitney U检验。通过计算Youden指数来确定RANTES水平的最佳临界值,以鉴别结直肠癌患者。用二元Logistic回归模型结合血清/血浆生物标志物的诊断效果进行拟合。所有统计检验均为双侧检验,差异有统计学意义(P<0.05)。结果与健康者相比,结直肠癌患者血浆RANTES显著高表达,差异有统计学意义(P<0.001)。RANTES的ROC曲线下面积为0.846(95%CI:0.791~0.901);RANTES和CEA联合检测AUC为0.891(95%CI:0.854~0.929),灵敏度和特异度分别达到0.854和0.803。结论RANTES对结直肠癌有诊断价值,RANTES联合CEA检测较单独检测CEA有更高的结直肠癌诊断效能。
文摘An expected 276bp fragment of the gene precursor encoding the signal peptide and mature protein of human β chemokine RANTES was amplified by reverse transcription\|polymerase chain reaction (RT\|PCR) from RNA of PHA\|activated human peripheral blood lymphocytes.This putative interested gene was inserted directly into a T\|vector and the ligation was confirmed by restriction enzyme digestion.The sequence data of the cloned fragment showed that it was almost identical with published sequences of RANTES gene,except for only one nucleotide substitution within the signal peptide region.The \%in vitro\% expressed recombinant RANTES protein was detected by the chemiluminescence enzyme\|linked immune Dot blotting assay after combining the recombinant plasmid with the \%in vitro\% SP6/T7 transcription and translation system.The successful cloning and expression of RANTES gene should shed light on future′s gene therapy of AIDS.
