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
目的:探讨高糖环境中人近端肾小管上皮细胞(HK-2)RANTES(regulated upon activation normal Tcell expressed and secreted)和转化生长因子β1(TGF-β1)表达变化及辛伐他汀(Sim)的干预作用。方法:原代培养HK-2分为正常糖对照组(NG)、甘...目的:探讨高糖环境中人近端肾小管上皮细胞(HK-2)RANTES(regulated upon activation normal Tcell expressed and secreted)和转化生长因子β1(TGF-β1)表达变化及辛伐他汀(Sim)的干预作用。方法:原代培养HK-2分为正常糖对照组(NG)、甘露醇组(MG)、高糖组(HG)、HG+Sim2μmol/L干预组(HG+Sim2)、HG+Sim4μmol/L干预组(HG+Sim4),RT-PCR法测定细胞RANTES和TGF-β1mRNA表达,ELISA法检测细胞上清液中RANTES、TGF-β1蛋白含量。结果:RANTES、TGF-β1的mRNA和蛋白在NG及MG仅有少量表达,而在HG则明显升高(P<0.05);加入Sim后,上述表达均较前下降(P<0.05),具有浓度依赖性。各组HK-2表达的RANTES和TGF-β1无论是mRNA还是蛋白之间均存在显著正相关(P<0.01)。结论:高糖促进HK-2RANTES、TGF-β1的表达,而Sim明显抑制高糖环境中上述因子的过度表达,发挥肾脏保护作用。展开更多
CCL5/RANTES（regulated upon activation,normal T-cell expressed and secreted）是一种细胞趋化因子,研究证明其与银屑病的发病相关[1-3]。紫外线照射是一种有效的银屑病治疗方法,但是,关于紫外线与CCL5/RANTES的相关性还不清楚。笔...CCL5/RANTES（regulated upon activation,normal T-cell expressed and secreted）是一种细胞趋化因子,研究证明其与银屑病的发病相关[1-3]。紫外线照射是一种有效的银屑病治疗方法,但是,关于紫外线与CCL5/RANTES的相关性还不清楚。笔者采用酶联免疫吸附法（ELISA）测定紫外线治疗前、后银屑病皮损角质形成细胞培养上清液中CCL5/RANTES含量,探讨紫外线治疗与银屑病皮损表皮中CCL5/RANTES含量的关系。展开更多
目的研究呼吸道合胞病毒(RSV)感染对哮喘小鼠RANTES(regulated upon activation normal T cellexpressed and secreted)生成的影响。方法将BALB/c小鼠分为对照组、哮喘组、RSV感染组和RSV感染哮喘组4组,每组各8只。先采用卵清白蛋白致...目的研究呼吸道合胞病毒(RSV)感染对哮喘小鼠RANTES(regulated upon activation normal T cellexpressed and secreted)生成的影响。方法将BALB/c小鼠分为对照组、哮喘组、RSV感染组和RSV感染哮喘组4组,每组各8只。先采用卵清白蛋白致敏法制作小鼠哮喘模型,随后以RSV感染小鼠,RSV感染后24 h处死小鼠。检测小鼠支气管肺泡灌洗液(BALF)中RANTES及IFN-γ的浓度,并计数炎性细胞数。结果 RSV感染组和对照组,RSV感染哮喘组和哮喘组小鼠之间,BALF中的RANTES浓度差异均有统计学意义(P均<0.05);而对照组和哮喘组,RSV感染组和RSV感染哮喘组小鼠之间的RANTES浓度差异则无统计学意义(P均>0.05)。各组小鼠BALF中的IFN-γ浓度差异无统计学意义。哮喘组、RSV感染组、RSV感染哮喘组小鼠BALF中的各种炎性细胞计数均较对照组升高,差异均有统计学意义(P<0.05),哮喘小鼠以嗜酸性粒细胞增高为主,而RSV感染小鼠以中性粒细胞为主。结论感染RSV可使小鼠RANTES的表达增加,但哮喘未能使小鼠RANTES表达增加,提示RANTES增加可能与RSV感染有关,而与哮喘状态无关。展开更多
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.
文摘目的:探讨高糖环境中人近端肾小管上皮细胞(HK-2)RANTES(regulated upon activation normal Tcell expressed and secreted)和转化生长因子β1(TGF-β1)表达变化及辛伐他汀(Sim)的干预作用。方法:原代培养HK-2分为正常糖对照组(NG)、甘露醇组(MG)、高糖组(HG)、HG+Sim2μmol/L干预组(HG+Sim2)、HG+Sim4μmol/L干预组(HG+Sim4),RT-PCR法测定细胞RANTES和TGF-β1mRNA表达,ELISA法检测细胞上清液中RANTES、TGF-β1蛋白含量。结果:RANTES、TGF-β1的mRNA和蛋白在NG及MG仅有少量表达,而在HG则明显升高(P<0.05);加入Sim后,上述表达均较前下降(P<0.05),具有浓度依赖性。各组HK-2表达的RANTES和TGF-β1无论是mRNA还是蛋白之间均存在显著正相关(P<0.01)。结论:高糖促进HK-2RANTES、TGF-β1的表达,而Sim明显抑制高糖环境中上述因子的过度表达,发挥肾脏保护作用。
文摘CCL5/RANTES（regulated upon activation,normal T-cell expressed and secreted）是一种细胞趋化因子,研究证明其与银屑病的发病相关[1-3]。紫外线照射是一种有效的银屑病治疗方法,但是,关于紫外线与CCL5/RANTES的相关性还不清楚。笔者采用酶联免疫吸附法（ELISA）测定紫外线治疗前、后银屑病皮损角质形成细胞培养上清液中CCL5/RANTES含量,探讨紫外线治疗与银屑病皮损表皮中CCL5/RANTES含量的关系。
文摘目的研究呼吸道合胞病毒(RSV)感染对哮喘小鼠RANTES(regulated upon activation normal T cellexpressed and secreted)生成的影响。方法将BALB/c小鼠分为对照组、哮喘组、RSV感染组和RSV感染哮喘组4组,每组各8只。先采用卵清白蛋白致敏法制作小鼠哮喘模型,随后以RSV感染小鼠,RSV感染后24 h处死小鼠。检测小鼠支气管肺泡灌洗液(BALF)中RANTES及IFN-γ的浓度,并计数炎性细胞数。结果 RSV感染组和对照组,RSV感染哮喘组和哮喘组小鼠之间,BALF中的RANTES浓度差异均有统计学意义(P均<0.05);而对照组和哮喘组,RSV感染组和RSV感染哮喘组小鼠之间的RANTES浓度差异则无统计学意义(P均>0.05)。各组小鼠BALF中的IFN-γ浓度差异无统计学意义。哮喘组、RSV感染组、RSV感染哮喘组小鼠BALF中的各种炎性细胞计数均较对照组升高,差异均有统计学意义(P<0.05),哮喘小鼠以嗜酸性粒细胞增高为主,而RSV感染小鼠以中性粒细胞为主。结论感染RSV可使小鼠RANTES的表达增加,但哮喘未能使小鼠RANTES表达增加,提示RANTES增加可能与RSV感染有关,而与哮喘状态无关。
