Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located...Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located in ZIKV 5'UTR and virus production.Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining,Subsequently,liquid chromatographytandem mass spectrometry(LC-MS/MS),bioinformatics analysis,and Western blot were used to identify the candidate proteins binding to ZIKV 5'UTR.Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production,respecitvely.Results tRSA RNA pull-down assay,LC-MS/MS,and Western blot analysis showed that polypyrimidine tractbinding protein(PTB)bound to the ZIKV 5'UTR.Furthermore,dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV(t=10.220,P<0.001),while PTB knockdown had the opposite effect(t=4.897,P<0.01).Additionally,virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer(t=6.400,P<0.01),whereas reducing PTB expression level weakened virus infectivity(t=5.055,P<0.01).Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.展开更多
一、青年报告Development of A New Animal Model of Coronary Microembolization Under Transthoracic Echocardiographic Guidance in RatsFAN Man-lu^(1),TONG Hua-qin^(2),SUN Tong^(3),DING Yu-han^(3),WATNG Meng-xi^(3),XIANG Qi...一、青年报告Development of A New Animal Model of Coronary Microembolization Under Transthoracic Echocardiographic Guidance in RatsFAN Man-lu^(1),TONG Hua-qin^(2),SUN Tong^(3),DING Yu-han^(3),WATNG Meng-xi^(3),XIANG Qian^(3),ZHANG Hao-wen^(*),CHEN Jian-dong^(*),CHEN Xiao-hu^(*)1.Department of TCM,The First Affiliated Hospital of Shandong First Medical University,Shandong Provincial Qianfoshan Hospital,Jinan,250000,China2.Department of Cardiology,Yangzhou Hospital of Chinese Medicine,Yangzhou,225000,China3.First College of Clinical Medicine,Nanjing University of Chinese Medicine,Nanjing,210023,China[ABSTRACT] Objective:There were multiple ways to establish coronary micro-embolization(CME)in animals to mimic human disease progression and evaluate treatment efficiency,using large animal models (dog and pig) with the interventional operation or small animal models (like the rat) requiring thoracotomy or custom-made catheter under fluoroscopic guidance.展开更多
文摘Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located in ZIKV 5'UTR and virus production.Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining,Subsequently,liquid chromatographytandem mass spectrometry(LC-MS/MS),bioinformatics analysis,and Western blot were used to identify the candidate proteins binding to ZIKV 5'UTR.Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production,respecitvely.Results tRSA RNA pull-down assay,LC-MS/MS,and Western blot analysis showed that polypyrimidine tractbinding protein(PTB)bound to the ZIKV 5'UTR.Furthermore,dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV(t=10.220,P<0.001),while PTB knockdown had the opposite effect(t=4.897,P<0.01).Additionally,virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer(t=6.400,P<0.01),whereas reducing PTB expression level weakened virus infectivity(t=5.055,P<0.01).Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.
文摘一、青年报告Development of A New Animal Model of Coronary Microembolization Under Transthoracic Echocardiographic Guidance in RatsFAN Man-lu^(1),TONG Hua-qin^(2),SUN Tong^(3),DING Yu-han^(3),WATNG Meng-xi^(3),XIANG Qian^(3),ZHANG Hao-wen^(*),CHEN Jian-dong^(*),CHEN Xiao-hu^(*)1.Department of TCM,The First Affiliated Hospital of Shandong First Medical University,Shandong Provincial Qianfoshan Hospital,Jinan,250000,China2.Department of Cardiology,Yangzhou Hospital of Chinese Medicine,Yangzhou,225000,China3.First College of Clinical Medicine,Nanjing University of Chinese Medicine,Nanjing,210023,China[ABSTRACT] Objective:There were multiple ways to establish coronary micro-embolization(CME)in animals to mimic human disease progression and evaluate treatment efficiency,using large animal models (dog and pig) with the interventional operation or small animal models (like the rat) requiring thoracotomy or custom-made catheter under fluoroscopic guidance.
