Objective:To study evolutionary relationship of the 5'untranslated regions(5'UTRs) in low passage dengue3 viruses(DEN3) isolated from hospitalized children with different clinical manifestations in Bangkok dur...Objective:To study evolutionary relationship of the 5'untranslated regions(5'UTRs) in low passage dengue3 viruses(DEN3) isolated from hospitalized children with different clinical manifestations in Bangkok during 24 year-evolution(1977-2000) comparing to the DEN3prototype(H87).Methods:The 5'UTRs of these Thai DEN3 and the H87 prototype were amplified by RT-PCR and sequenced.Their multiple sequence alignments were done by Codon Code Aligner v 4.0.4 software and their RNA secondary structures were predicted by MFOLD software.Replication of five Thai DEN3 candidates comparing to the 1187 prototype were done in human(HepG2) and the mosquito(C6/36) cell lines.Results:Among these Thai DEN3,the completely identical sequences of their first 89 nucleotides,their high-order secondary structure of 5'UTRs and three SNPs including the predominant C90 T,and two minor SNPs including A109 G and A112 G were found.The C90 T of Thai DEN3.Bangkok isolates was shown predominantly before 1977.Five Thai DEN3 candidates with the predominant C90 T were shown to replicate in human(HepG2) and the mosquito(C6/36) cell lines better than the H87 prototype.However,their highly conserved sequences as well as SNPs of the 5'UTR did not appear to correlate with their disease severity in human.Conclusions:Our findings highlighted evolutionary relationship of the completely identical 89 nucleotide sequence,the high-order secondary structure and the predominant C90 T of the 5'UTR of these Thai DEN3 during 24 year-evolution further suggesting to be their genetic markers and magic targets for future research on antiviral therapy as well as vaccine approaches of Thai DEN3.展开更多
目的:探讨三结构域蛋白家族24(tripartite motif-containing protein 24,TRIM24)、c-myc及ARF蛋白在食管鳞状细胞癌组织中的表达及其意义。方法:选取2018年01月至2022年12月期间承德市中心医院接受手术治疗的102例食管鳞癌患者临床病理...目的:探讨三结构域蛋白家族24(tripartite motif-containing protein 24,TRIM24)、c-myc及ARF蛋白在食管鳞状细胞癌组织中的表达及其意义。方法:选取2018年01月至2022年12月期间承德市中心医院接受手术治疗的102例食管鳞癌患者临床病理资料作为研究对象(实验组),同时选取癌旁正常食管黏膜组织54例(对照组),采用免疫组化(Envision法)对TRIM24、c-myc及ARF蛋白进行检测,观察三者在鳞癌组织及癌旁正常食管黏膜组织中的表达情况,并分析三者的表达与临床病理特征的关系,同时研究三者于食管鳞癌组织内表达的相关性。结果:TRIM24、c-myc及ARF在鳞癌组织及癌旁正常食管黏膜组织中阳性表达率分别为72.62%vs 7.45%、74.36%vs 13.85%、26.56%vs 83.18%,三种蛋白在鳞癌组织及正常食管黏膜组织中的表达差异具有统计学意义(P均<0.05);经分析,食管鳞癌组织中的TRIM24、c-myc及ARF蛋白表达与癌的分化程度、淋巴结转移、临床分期具有显著相关性(P均<0.05);经Spearman相关性分析,食管鳞癌组织中TRIM24与c-myc蛋白表达呈正相关(r=0.587,P<0.05),TRIM24与ARF蛋白表达呈负相关(r=-0.601,P<0.05),c-myc与ARF蛋白表达呈负相关(r=-0.465,P<0.05)。结论:TRIM24、c-myc、ARF的异常表达可能对食管鳞状细胞癌的发生及发展有一定关系。因此,c-myc、TRIM24和ARF可能作为新的潜在治疗靶点,为食管鳞癌患者的个体化治疗提供新的方向。展开更多
基金supported by two research grants of Associate Professor Dr.W.Attatippaholkun:Grant No.493-5600-G-00-3461,Program in Science and Technology Cooperation,Human Capacity Development,Bureau for Global Programs,Field Support and Research,US Agency for International Development,Washington,DCThe Royal Golden Jubilee-Ph.D Program,Thailand Research Fund,Thailand
文摘Objective:To study evolutionary relationship of the 5'untranslated regions(5'UTRs) in low passage dengue3 viruses(DEN3) isolated from hospitalized children with different clinical manifestations in Bangkok during 24 year-evolution(1977-2000) comparing to the DEN3prototype(H87).Methods:The 5'UTRs of these Thai DEN3 and the H87 prototype were amplified by RT-PCR and sequenced.Their multiple sequence alignments were done by Codon Code Aligner v 4.0.4 software and their RNA secondary structures were predicted by MFOLD software.Replication of five Thai DEN3 candidates comparing to the 1187 prototype were done in human(HepG2) and the mosquito(C6/36) cell lines.Results:Among these Thai DEN3,the completely identical sequences of their first 89 nucleotides,their high-order secondary structure of 5'UTRs and three SNPs including the predominant C90 T,and two minor SNPs including A109 G and A112 G were found.The C90 T of Thai DEN3.Bangkok isolates was shown predominantly before 1977.Five Thai DEN3 candidates with the predominant C90 T were shown to replicate in human(HepG2) and the mosquito(C6/36) cell lines better than the H87 prototype.However,their highly conserved sequences as well as SNPs of the 5'UTR did not appear to correlate with their disease severity in human.Conclusions:Our findings highlighted evolutionary relationship of the completely identical 89 nucleotide sequence,the high-order secondary structure and the predominant C90 T of the 5'UTR of these Thai DEN3 during 24 year-evolution further suggesting to be their genetic markers and magic targets for future research on antiviral therapy as well as vaccine approaches of Thai DEN3.
