Objective： To construct recombinant adeno-associated virus type 2（rAAV2） vectors encoding human apoA-I/apoA-lMilano protein, and explore an effective and safe method to prevent and treat the atherosclerotic disease...Objective： To construct recombinant adeno-associated virus type 2（rAAV2） vectors encoding human apoA-I/apoA-lMilano protein, and explore an effective and safe method to prevent and treat the atherosclerotic diseases. Methods： Human apoA-I cDNA with a His-tag in the upward stream of cDNA sequence were obtained with RT-PCR and PCR, and human apoA-IMilano cDNA was prepared by QuikChange Site-Directed Mutagenesis Kit. After extracted rAAV vectors with a most economic and convenient method, the particle numbers of rAAV vectors were measured by Dot-blot, and the purity was assayed by SOS-Page. The expression efficiency of the apoA-I or apoA-IMilano in C2C12 infected by rAAV vectors were detected by ELISA method. Results： ApoA-I cDNA was gained by RT-PCR and a His-tag was added in the upward stream of apoA-I cDNA successfully. ApoA-I cDNA was mutanted to apoA-IMilano cDNA successfully by QuikChange Site-Directed Mutagenesis Kit. The both titres of the rAAV vectors of apoA-I and apoA-IMilano were about 2×10^14/L, and the result of SOS-Page showed that the purity of the rAAV vectors was satisfied. The expression level of apoA-I was （0.39±0.04） μg/ml and the apoA-IMilano was （0.31±0.03） μ/ml in the DMEM culture medium at the first 24h after transfection. Conclusion： The success of the rAAV vectors construction, purification and the expression of apoA-I and apoA-IMilano in C2C12 cells mediated by these vectors, makes possible to inject rAAVA and rAAVAM vectors into mice muscle, and rises a new hope on finding a new way to prevent and treat atherosclerotic diseases and cardiovascular disease.展开更多
Objective:To study the anti-respiratory syncytial viruses(RSV) mechanism of extracts from Hibiscus mutabilis leaf in vitro.Methods:Elibavirin was used as the positive drug to detect the anti-RSV activity of extracts f...Objective:To study the anti-respiratory syncytial viruses(RSV) mechanism of extracts from Hibiscus mutabilis leaf in vitro.Methods:Elibavirin was used as the positive drug to detect the anti-RSV activity of extracts from Hibiscus mutabilis leaf. By observing the antiviral effect of the extracts of Hibiscus mutabilis leaf in different ways of administration, the anti-RSV mechanism of the extracts of Hibiscus mutabilis leaf was preliminarily explored.Results:Hibiscus mutabilis leaf extracts could effectively inhibit the cytopathic changes caused by RSV, and had significant antiviral effect. The therapeutic index (TI) was 86.24.Conclusion:Hibiscus mutabilis leaf extracts have significant anti-RSV effect in vitro, and their antiviral effect occurs after virus penetration.展开更多
文摘Objective： To construct recombinant adeno-associated virus type 2（rAAV2） vectors encoding human apoA-I/apoA-lMilano protein, and explore an effective and safe method to prevent and treat the atherosclerotic diseases. Methods： Human apoA-I cDNA with a His-tag in the upward stream of cDNA sequence were obtained with RT-PCR and PCR, and human apoA-IMilano cDNA was prepared by QuikChange Site-Directed Mutagenesis Kit. After extracted rAAV vectors with a most economic and convenient method, the particle numbers of rAAV vectors were measured by Dot-blot, and the purity was assayed by SOS-Page. The expression efficiency of the apoA-I or apoA-IMilano in C2C12 infected by rAAV vectors were detected by ELISA method. Results： ApoA-I cDNA was gained by RT-PCR and a His-tag was added in the upward stream of apoA-I cDNA successfully. ApoA-I cDNA was mutanted to apoA-IMilano cDNA successfully by QuikChange Site-Directed Mutagenesis Kit. The both titres of the rAAV vectors of apoA-I and apoA-IMilano were about 2×10^14/L, and the result of SOS-Page showed that the purity of the rAAV vectors was satisfied. The expression level of apoA-I was （0.39±0.04） μg/ml and the apoA-IMilano was （0.31±0.03） μ/ml in the DMEM culture medium at the first 24h after transfection. Conclusion： The success of the rAAV vectors construction, purification and the expression of apoA-I and apoA-IMilano in C2C12 cells mediated by these vectors, makes possible to inject rAAVA and rAAVAM vectors into mice muscle, and rises a new hope on finding a new way to prevent and treat atherosclerotic diseases and cardiovascular disease.
基金Natural science funds of Shandong province(ZR2016HL53).
文摘Objective:To study the anti-respiratory syncytial viruses(RSV) mechanism of extracts from Hibiscus mutabilis leaf in vitro.Methods:Elibavirin was used as the positive drug to detect the anti-RSV activity of extracts from Hibiscus mutabilis leaf. By observing the antiviral effect of the extracts of Hibiscus mutabilis leaf in different ways of administration, the anti-RSV mechanism of the extracts of Hibiscus mutabilis leaf was preliminarily explored.Results:Hibiscus mutabilis leaf extracts could effectively inhibit the cytopathic changes caused by RSV, and had significant antiviral effect. The therapeutic index (TI) was 86.24.Conclusion:Hibiscus mutabilis leaf extracts have significant anti-RSV effect in vitro, and their antiviral effect occurs after virus penetration.
