The expression of therapeutic gene and its anti-tumor effects will beaugmented and a synergism of oncolytic virus with the therapeutic gene is speculated. This study wasundertaken to assess the anti-tumor effects of a...The expression of therapeutic gene and its anti-tumor effects will beaugmented and a synergism of oncolytic virus with the therapeutic gene is speculated. This study wasundertaken to assess the anti-tumor effects of a novel gene-viral therapeutic systemCNHK300-mEndostatin ( CNHK300-mE) in hepatocellular carcinoma (HCC). A novel gene-viral therapeuticsystem named CNHK300-mE was constructed using the human telomerase reverse transcriptase (hTERT)promoter to drive the expression of the adenovirus El A gene and cloning the therapeutic gene mouseendostatin into the adenovirus genome. By the tissue culture infectious dose 50 (TCID50) method andcytoviability assay, the replicative and cytolytic capabilities of CNHK300-mE in two HCC lines (HepGII and Hep3B) and one normal cell line ( MRC-5 ) were analyzed, and the transgene expressions ofmouse endostatin in vitro and in vivo were detected by Western blotting and ELISA assay. Tumorgrowth suppression and anti-angiogenesis effects in vivo were investigated using nude micexenografts model derived from SMMC-7721 HCC cells. The 3296-fold replicating capacity of CNHK300-mEin HCC cell lines versus in the normal cell line at 96 hours post infection and the 25 -foldeffective dose for killing 50% cells ( ED50) in the normal cell line versus HCC cell lines, whichwere both superior to ONYX-015, were observed. Tumor growth suppression of CNHK300-mE superior toeither Ad-mE or ONYX-015 was demonstrated (P < 0. 01) and the anti-angiogenic effects in vivosuperior to Ad-mE were also observed with immunohistochemical staining of von Willebrand factor. Incomparison with non-replicative adenovirus Ad-mE, the transgene expression of mE mediated byCNHK300- in vivo (P<0.05). Being capable of replicating in and lysing the telomerase-positive HCCcells and mediating effective expression of the therapeutic gene in vitro and in vivo, the novelgene-viral therapeutic system CNHK300-mE is potentially effective in the treatment of HCC.展开更多
适配体(Aptamers)是通过指数富集的配体系统进化(systematic evolution of ligands by exponential enrichment,SELEX)技术,从随机核酸文库中筛选出来的单链寡核苷酸,已在临床医疗及其他领域得到日益广泛的应用.与抗体相比,适配体具有...适配体(Aptamers)是通过指数富集的配体系统进化(systematic evolution of ligands by exponential enrichment,SELEX)技术,从随机核酸文库中筛选出来的单链寡核苷酸,已在临床医疗及其他领域得到日益广泛的应用.与抗体相比,适配体具有很多优点,如高亲和力、高特异性、分子量小、几乎无免疫排斥反应、结构稳定、易于合成等.可用于适配体筛选的靶标范围非常广,包括有机小分子、蛋白质、完整细胞及病毒颗粒等.迅速可靠的病原检测对于病毒性传染病的成功预防和治疗具有重要意义.随着严格筛选和快速分离技术的进步,适配体在病毒感染的检测治疗中显示出巨大的潜力.本文概括介绍了适配体在病毒研究方面的最新应用进展及未来前景.展开更多
BACKGROUND: Much evidence demonstrates that the genotypes of hepatitis B virus (HBV) present differences in pathogenicity and outcomes owing to differences in genetic structure. This study aimed to investigate the inf...BACKGROUND: Much evidence demonstrates that the genotypes of hepatitis B virus (HBV) present differences in pathogenicity and outcomes owing to differences in genetic structure. This study aimed to investigate the influences of HBV genotypes on the anti-viral therapeutic efficacy of interferon-alpha (IFN-alpha) in chronic hepatitis B patients, and to determine the relationship between HBV genotypes and levels of viral replication or gene variations. METHODS: The chronic hepatitis B patients who were treated with IFN-alpha were selected randomly. Anti-viral therapeutic efficacy was monitored in these patients. The HBV genotypes were detected by PCR microplate hybridization ELISA. The levels of serum HBV-DNA were determined by fluorescence quantitative PCR. HBV gene variation at pre-C and basic core promoter (BCP) regions were assayed by gene chip technology. RESULTS: Genotypes B and C were predominant in 94 chronic hepatitis B patients. A, E and F genotypes were not found in these patients. The HBV-DNA levels of genotype C and mixed genotypes were significantly higher than those of genotype B. The response to IFN-alpha in patients with genotype B was markedly better than in those with genotypes C and D, and the complete response to IFN-alpha was only observed in genotype B. The response to IFN-alpha in patients with mixed genotypes was the least sensitive. The negative transition of HBeAg was correlated with variations in the HBV pre-C and BCP regions in patients with partial or no response to IFN-alpha. The variation rates of HBV pre-C and BCP regions were clearly higher in genotype C than in genotype B. CONCLUSIONS: The results suggest that HBV genotype is correlated with the serum levels of HBV-DNA, HBV gene variations and therapeutic efficacy of IFN-alpha. The regular detection of HBV genotypes in the clinic will be of benefit for disease prognosis and planning of anti-viral therapeutic strategies.展开更多
文摘The expression of therapeutic gene and its anti-tumor effects will beaugmented and a synergism of oncolytic virus with the therapeutic gene is speculated. This study wasundertaken to assess the anti-tumor effects of a novel gene-viral therapeutic systemCNHK300-mEndostatin ( CNHK300-mE) in hepatocellular carcinoma (HCC). A novel gene-viral therapeuticsystem named CNHK300-mE was constructed using the human telomerase reverse transcriptase (hTERT)promoter to drive the expression of the adenovirus El A gene and cloning the therapeutic gene mouseendostatin into the adenovirus genome. By the tissue culture infectious dose 50 (TCID50) method andcytoviability assay, the replicative and cytolytic capabilities of CNHK300-mE in two HCC lines (HepGII and Hep3B) and one normal cell line ( MRC-5 ) were analyzed, and the transgene expressions ofmouse endostatin in vitro and in vivo were detected by Western blotting and ELISA assay. Tumorgrowth suppression and anti-angiogenesis effects in vivo were investigated using nude micexenografts model derived from SMMC-7721 HCC cells. The 3296-fold replicating capacity of CNHK300-mEin HCC cell lines versus in the normal cell line at 96 hours post infection and the 25 -foldeffective dose for killing 50% cells ( ED50) in the normal cell line versus HCC cell lines, whichwere both superior to ONYX-015, were observed. Tumor growth suppression of CNHK300-mE superior toeither Ad-mE or ONYX-015 was demonstrated (P < 0. 01) and the anti-angiogenic effects in vivosuperior to Ad-mE were also observed with immunohistochemical staining of von Willebrand factor. Incomparison with non-replicative adenovirus Ad-mE, the transgene expression of mE mediated byCNHK300- in vivo (P<0.05). Being capable of replicating in and lysing the telomerase-positive HCCcells and mediating effective expression of the therapeutic gene in vitro and in vivo, the novelgene-viral therapeutic system CNHK300-mE is potentially effective in the treatment of HCC.
文摘适配体(Aptamers)是通过指数富集的配体系统进化(systematic evolution of ligands by exponential enrichment,SELEX)技术,从随机核酸文库中筛选出来的单链寡核苷酸,已在临床医疗及其他领域得到日益广泛的应用.与抗体相比,适配体具有很多优点,如高亲和力、高特异性、分子量小、几乎无免疫排斥反应、结构稳定、易于合成等.可用于适配体筛选的靶标范围非常广,包括有机小分子、蛋白质、完整细胞及病毒颗粒等.迅速可靠的病原检测对于病毒性传染病的成功预防和治疗具有重要意义.随着严格筛选和快速分离技术的进步,适配体在病毒感染的检测治疗中显示出巨大的潜力.本文概括介绍了适配体在病毒研究方面的最新应用进展及未来前景.
基金This study was supported by a grant from the Scientific and Technology Bureau of Hubei Province Foundation(No.2005AA301C26).
文摘BACKGROUND: Much evidence demonstrates that the genotypes of hepatitis B virus (HBV) present differences in pathogenicity and outcomes owing to differences in genetic structure. This study aimed to investigate the influences of HBV genotypes on the anti-viral therapeutic efficacy of interferon-alpha (IFN-alpha) in chronic hepatitis B patients, and to determine the relationship between HBV genotypes and levels of viral replication or gene variations. METHODS: The chronic hepatitis B patients who were treated with IFN-alpha were selected randomly. Anti-viral therapeutic efficacy was monitored in these patients. The HBV genotypes were detected by PCR microplate hybridization ELISA. The levels of serum HBV-DNA were determined by fluorescence quantitative PCR. HBV gene variation at pre-C and basic core promoter (BCP) regions were assayed by gene chip technology. RESULTS: Genotypes B and C were predominant in 94 chronic hepatitis B patients. A, E and F genotypes were not found in these patients. The HBV-DNA levels of genotype C and mixed genotypes were significantly higher than those of genotype B. The response to IFN-alpha in patients with genotype B was markedly better than in those with genotypes C and D, and the complete response to IFN-alpha was only observed in genotype B. The response to IFN-alpha in patients with mixed genotypes was the least sensitive. The negative transition of HBeAg was correlated with variations in the HBV pre-C and BCP regions in patients with partial or no response to IFN-alpha. The variation rates of HBV pre-C and BCP regions were clearly higher in genotype C than in genotype B. CONCLUSIONS: The results suggest that HBV genotype is correlated with the serum levels of HBV-DNA, HBV gene variations and therapeutic efficacy of IFN-alpha. The regular detection of HBV genotypes in the clinic will be of benefit for disease prognosis and planning of anti-viral therapeutic strategies.
