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1Torque teno virus:Its prevalence and isotypes in North India显示文摘AIM: To investigate the prevalence and genotype dis-tribution of Torque teno virus (TTV) in patients with different liver diseases and chronic renal failure treated at a referral hospital in North India. METHODS: Whereas prevalence of TTV was based on amplification of conserved region of ORF2 of TTV genome,the genotyping of TTV was carried out using restriction fragment length polymorphism (RFLP) procedure on the N22 region of ORF1. RESULTS: TTV-DNA was detected in 137 of 513 (26.7%) patients with liver diseases and 38 of 65 (58.5%) patients with chronic renal failure. TTV was also detected in 27% of healthy controls. The sequence analysis of the PCR product from 10 randomly selected cases failed to show a significant sequence divergence when compared with that of the TRM1 isolate of TTV genotype 1. The results of genotyping in 55 randomly selected patients showed the presence of genotype 1 (G1) in 53 (96.4%) and genotype 2 (G2) in 2 cases (3.6%),respectively. Other genotypes were not identified in this patient subgroup,suggesting that G1 is predominant in this area. The results of genotyping by RFLP were also supported by phylogenetic tree analysis,where G1 was found to be the major genotype. CONCLUSION: These results indicate that TTV is moderately present in Indian patients,with G1 to be the major genotype in North India. The pathogenicity and etiological role of TTV in different diseases is still a question mark and warrant further studies.Mohammad Irshad Shiwani Singh Khushboo Irshad Sanjay Kumar Agarwal Yogendra Kumar Joshi 2008World Journal of Gastroenterology2008,14,39:5
2Prevalence and clinical significance of SEN virus infection in patients with non A-E hepatitis and volunteer blood donors in Shanghai显示文摘AIM:To explore the prevalence of SEN virus (SENV) in patients with non A-E hepatitis and volunteer blood donors in Shanghai. METHODS: According to the published gene sequences, primers from the conserved region were designed. Then, the prevalence of SEN virus in 30 samples from healthy voluntary blood donors and 30 samples from patients with non A-E hepatitis were detected by nested-PCR of SENV-D/H. Some PCR products were cloned and sequenced. RESULTS: The specificity of genotype-specific PCR was confirmed by sequencing, the SENV DNA was detected in 53.3% of the patients with non A-E hepatitis and 10% of the blood donors. The prevalence of SENV-D/H viremia was significantly higher in patients with non A-E hepatitis than in blood donors (P = 0.0002). SENV-H subtype and SENV-D subtype were found in 2 and 1 samples, respectively from blood donors. SENV-H subtype, SENV D subtype, mixed SENV-D and SENV-H subtype were found in 8, 6 and 2 samples, respectively, from patients with non A-E hepatitis. CONCLUSION: The gene type of SENV in patients with non A-E hepatitis and blood donors in shanghai is D or H subtype, and transfusion is not the only transmitting form of SENV.Zheng-Hao Tang Xiao-Hua Chen Yong-Sheng Yu Guo-Qing Zang 2008World Journal of Gastroenterology2008,14,26:0
3隐源性肝炎相关新基因CHBP2的原核表达及蛋白纯化显示文摘目的:构建隐源性肝炎相关新基因CHBP2原核表达载体,在大肠杆菌中进行表达,并纯化CHBP2融合蛋白。方法:应用逆转录聚合酶链反应(RT-PCR),以提取的Huh7细胞mRNA为模板,扩增获得CHBP2基因片段,连接到pGEM-T载体,测序正确后插入至原核表达载体pET-32a(+)中,构建原核表达载体pET-32a(+)-CHBP2,转化大肠杆菌BL21,以异丙基-β-D-硫代半乳糖苷(IPTG)诱导,获得CHBP2融合蛋白的可诱导性表达,通过SDS-PAGE电泳、Western blot免疫印迹分析证实蛋白表达的特异性,超声破碎表达细菌,SDS-PAGE分析,利用镍离子亲和柱对表达蛋白进行纯化及柱上复性。结果:成功构建原核表达载体pET-32a(+)- CHBP2,并将CHBP2融合蛋白成功表达,通过Western blot免疫印迹,证实了蛋白表达的特异性。SDS-PAGE分析表明其为包涵体表达,并对蛋白成功进行了纯化和复性,获得了表达蛋白的纯品结论:成功表达、纯化CHBP2蛋白,为研究CHBP2蛋白的生物学功能打下了基础,李锟 叶进 肖凡 李国力 洪源 魏红山 2007世界华人消化杂志2007,15,22:0
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