The inhibitory effect of parvovirus H-1 on the colonyforming ability in vitro of QGY-7703, a cultured human hepatoma cell line, and on the formation and growth of its tumors in nude mice was studied. With higher multi...The inhibitory effect of parvovirus H-1 on the colonyforming ability in vitro of QGY-7703, a cultured human hepatoma cell line, and on the formation and growth of its tumors in nude mice was studied. With higher multiplicity of infection (MOI) of H-1 given, survival of the QGY-7703 cells was found to be decreased. H-1 DNA amplification level at 30 h postinfection(p.i.) was detected to be 7.4 times higher than that at 2 h by dispersed cells assay, while the cells were delayed to enter into S phase.Plaques were formed in the indicator cells (new-born human kidney cell line, NBK) by progeny H-1 virus particles released from the infected QGY-7703 cells by infectious cell center assay. The formation of tumors in nude mice by QGY-7703 cells which were injected s c at 2 h postinfection was observed to be prevented in 2 groups with given MOI 25 and 50. The tumor growth of MOI 10 group occurred at a lower exponential rate than that of control,after a 20 d latent period. It was evident that parvovirus H-1 exhibited a direct inhibitory effect on the formation and growth of human hepatoma cells in vivo as well as in vitro.展开更多
We had given evidences of cytocidal effect in vitro of either parvovirus H-I(PV H-1) or AT-1840 (lycobetaine) on gastric cancer cells. They act on different phase of the cell cycle of the gastric cancer cells, with PV...We had given evidences of cytocidal effect in vitro of either parvovirus H-I(PV H-1) or AT-1840 (lycobetaine) on gastric cancer cells. They act on different phase of the cell cycle of the gastric cancer cells, with PV H-1 on the late stage of S phase and AT-1840 on M phase. Combined use of these drugs gives better killing effect than the individual drug alone. In gastric cancer-bearing nude mice (ascites and solid tumor forms), both AT-1840 and PV H-1 increased the survival peroids and decreased the size of solid tumors. Depsite these are the experimental study which may not to be the same in human being. However, the study has layed down a solid foundation for our further exploration of these two substances as anti-gastric cancer drugs in the future.展开更多
Parvovirus H-1 has been a promising vector in human gene therapy of tumor. With a vector, pSHll8, a mutation was performed down the site of p38 promoter to change ATG codon into ACG, which leaves the NS 1 protein inta...Parvovirus H-1 has been a promising vector in human gene therapy of tumor. With a vector, pSHll8, a mutation was performed down the site of p38 promoter to change ATG codon into ACG, which leaves the NS 1 protein intact and ensures correct expression of the downstream report genes, such as enhanced green fluorescent protein (GFP) gene and wt p53 cDNA, in newly constructed H1 based vectors (pH1EG and pHI/p53). The vector, pH1EG,展开更多
[Objective] The research aimed to provide the theoretical basis for establishing a rapid diagnosis method for porcine parvovirus（PPV）. [ Method] One pair of primers were designed according to PPV genome sequences on...[Objective] The research aimed to provide the theoretical basis for establishing a rapid diagnosis method for porcine parvovirus（PPV）. [ Method] One pair of primers were designed according to PPV genome sequences on GenBank website and the sequences of prokaryotic expression vector pET30a （ ＋ ） with multiple cloning sites. The whole sequence of NS1 gene in PPV SD1 strain was amplified by using PCR technology and the positive recombinant plasmid was analyzed by sequencing and homology comparison. The prokaryotic expression recombinant plasmid PET30a/NS1 was constructed to make its induction expression in Escherichia coll. [ Result] The target fragment with the length of 2 208 bp was obtained from PCR amplification. The nucleotide homologies between the cloned NS1 gene and the reported relevant PPV genes were from 97.3 % to 99.4 %, which indicated that NS1 gene had high conservation. But it had a 12-basepair successive deletion near the hydroxyl end. The cloned PPV NS1 gene was successfully expressed in prokaryotic cell, and its expression products existed mostly in inclusion bodies. [ Conclusion] The results of SDS-PAGE detection showed that the molecular weight of PPV NS1 protein was 86 KD.展开更多
Porcine parvovirus (PPV) is one of the major agents causing swine reproductive failure. NS1 protein is a non-structural protein of PPV and can be used as a reagent for differentiation of vaccinated animals and infecte...Porcine parvovirus (PPV) is one of the major agents causing swine reproductive failure. NS1 protein is a non-structural protein of PPV and can be used as a reagent for differentiation of vaccinated animals and infected ones. In present study, a recombinant plasmid pET28a/NS1 was constructed by cloning the coding sequence for NS1 of PPV into pET28a, a bacterial expression vector. The NS1 protein was expressed in E. coli BL21(DE3) after induced by IPTG and the recombinant fusion protein was purified with affinity chromatography. Expression amount of NS1 protein was improved by optimizing the inducing parameters. The recombinant NS1 protein is reactive to PPV positive sera in Western blot and ELISA test and therefore can be applicable in differential diagnosis of PPV infections.展开更多
文摘The inhibitory effect of parvovirus H-1 on the colonyforming ability in vitro of QGY-7703, a cultured human hepatoma cell line, and on the formation and growth of its tumors in nude mice was studied. With higher multiplicity of infection (MOI) of H-1 given, survival of the QGY-7703 cells was found to be decreased. H-1 DNA amplification level at 30 h postinfection(p.i.) was detected to be 7.4 times higher than that at 2 h by dispersed cells assay, while the cells were delayed to enter into S phase.Plaques were formed in the indicator cells (new-born human kidney cell line, NBK) by progeny H-1 virus particles released from the infected QGY-7703 cells by infectious cell center assay. The formation of tumors in nude mice by QGY-7703 cells which were injected s c at 2 h postinfection was observed to be prevented in 2 groups with given MOI 25 and 50. The tumor growth of MOI 10 group occurred at a lower exponential rate than that of control,after a 20 d latent period. It was evident that parvovirus H-1 exhibited a direct inhibitory effect on the formation and growth of human hepatoma cells in vivo as well as in vitro.
文摘We had given evidences of cytocidal effect in vitro of either parvovirus H-I(PV H-1) or AT-1840 (lycobetaine) on gastric cancer cells. They act on different phase of the cell cycle of the gastric cancer cells, with PV H-1 on the late stage of S phase and AT-1840 on M phase. Combined use of these drugs gives better killing effect than the individual drug alone. In gastric cancer-bearing nude mice (ascites and solid tumor forms), both AT-1840 and PV H-1 increased the survival peroids and decreased the size of solid tumors. Depsite these are the experimental study which may not to be the same in human being. However, the study has layed down a solid foundation for our further exploration of these two substances as anti-gastric cancer drugs in the future.
文摘Parvovirus H-1 has been a promising vector in human gene therapy of tumor. With a vector, pSHll8, a mutation was performed down the site of p38 promoter to change ATG codon into ACG, which leaves the NS 1 protein intact and ensures correct expression of the downstream report genes, such as enhanced green fluorescent protein (GFP) gene and wt p53 cDNA, in newly constructed H1 based vectors (pH1EG and pHI/p53). The vector, pH1EG,
基金Supported by the Natural Science Foundation Program of Shandong Province(2007ZRA16001)~~
文摘[Objective] The research aimed to provide the theoretical basis for establishing a rapid diagnosis method for porcine parvovirus（PPV）. [ Method] One pair of primers were designed according to PPV genome sequences on GenBank website and the sequences of prokaryotic expression vector pET30a （ ＋ ） with multiple cloning sites. The whole sequence of NS1 gene in PPV SD1 strain was amplified by using PCR technology and the positive recombinant plasmid was analyzed by sequencing and homology comparison. The prokaryotic expression recombinant plasmid PET30a/NS1 was constructed to make its induction expression in Escherichia coll. [ Result] The target fragment with the length of 2 208 bp was obtained from PCR amplification. The nucleotide homologies between the cloned NS1 gene and the reported relevant PPV genes were from 97.3 % to 99.4 %, which indicated that NS1 gene had high conservation. But it had a 12-basepair successive deletion near the hydroxyl end. The cloned PPV NS1 gene was successfully expressed in prokaryotic cell, and its expression products existed mostly in inclusion bodies. [ Conclusion] The results of SDS-PAGE detection showed that the molecular weight of PPV NS1 protein was 86 KD.
文摘Porcine parvovirus (PPV) is one of the major agents causing swine reproductive failure. NS1 protein is a non-structural protein of PPV and can be used as a reagent for differentiation of vaccinated animals and infected ones. In present study, a recombinant plasmid pET28a/NS1 was constructed by cloning the coding sequence for NS1 of PPV into pET28a, a bacterial expression vector. The NS1 protein was expressed in E. coli BL21(DE3) after induced by IPTG and the recombinant fusion protein was purified with affinity chromatography. Expression amount of NS1 protein was improved by optimizing the inducing parameters. The recombinant NS1 protein is reactive to PPV positive sera in Western blot and ELISA test and therefore can be applicable in differential diagnosis of PPV infections.
