AIM: To explore the inhibitory effects of pokeweed antiviral protein seed (PAP-S) and PAP encoded by a eukaryotic expression plasmid on hepatitis B virus (HBV) replication in vitro. METHODS: HepG2 2.2.15 cells in cult...AIM: To explore the inhibitory effects of pokeweed antiviral protein seed (PAP-S) and PAP encoded by a eukaryotic expression plasmid on hepatitis B virus (HBV) replication in vitro. METHODS: HepG2 2.2.15 cells in cultured medium were treated with different concentrations of PAP-S. HBsAg, HBeAg and HBV DNA in supernatants were determined by ELISA and fluorescent quantitative PCR respectively. MTT method was used to assay for cytotoxicity. HepG2 were cotransfected with various amounts of PAP encoded by a eukaryotic expression plasmid and replication competent wild-type HBV 1.3 fold over- length plasmid. On d 3 after transfection, HBsAg and HBeAg were determined by using ELISA. Levels of HBV core-associated DNA and RNA were detected by using Southern and Northern blot, respectively. RESULTS: The inhibitory effects of PAP-S on HBsAg, HBeAg and HBV DNA were gradually enhanced with the increase of PAP concentration. When the concentration of PAP-S was 10 μg/mL, the inhibition rates of HBsAg, HBeAg and HBV DNA were 20.9%, 30.2% and 50%, respectively. After transfection of 1.0 μg and 2.0 μg plasmid pXF3H-PAP, the levels of HBV nucleocapside- associated DNA were reduced by 38.0% and 74.0% respectively, the levels of HBsAg in the media by 76.8% and 99.7% respectively, and the levels of HBeAg by 72.7% and 99.3% respectively as compared with controls. Transfection with 2 μg plasmid pXF3H-PAP reduced the levels of HBV nucleocapside-associated RNA by 69.0%.CONCLUSION: Both PAP-S and PAP encoded by a eukaryotic expression plasmid could effectively inhibit HBV replication and antigen expression in vitro, and the inhibitory effects were dose-dependent.展开更多
To the Editor:Coronary heart disease(CHD)and depression are common in patients worldwide and have a high comorbidity rate.Researches on depression have shown that it has been linked with long-term risk for CHD.[1]The ...To the Editor:Coronary heart disease(CHD)and depression are common in patients worldwide and have a high comorbidity rate.Researches on depression have shown that it has been linked with long-term risk for CHD.[1]The incidence and mortality of depression with CHD were higher,and the prognosis was worse.[2]The association between CHD and depression is well established and is suggested to be bidirectional.[3]Therefore,to provide possible conditions for the study of depression with CHD in the future,establishing a reliable animal model of depression with CHD is necessary.Chronic unpredictable mild stress(CUMS)is the most widely used to establish an animal model of depression for the past few years.Feeding a high-fat diet(HFD)to rats is a widely used method for establishing a model for atherosclerosis and CHD.Research reported that female cynomolgus monkeys that were housed in single cages and consumed an atherogenic diet could develop depression combined with CHD model.[4]Another paper reported that CUMS combined with HFD in rats was similar to the clinical manifestation of vascular depression in humans.[5]The former demonstrated that housing in single cages and feeding an atherogenic diet in monkeys could establish a depression combined with CHD model,the latter demonstrated that CUMS combined with HFD in rats was similar to vascular depression.It remains unclear whether CUMS combined with HFD in rats could establish a complex rat model of depression with CHD.Consequently,the aim of this study was to explore whether CUMS combined with HFD in rats could establish a representative,reliable rat model of depression with CHD and to provide possible conditions for an in-depth study of depression with CHD.展开更多
基金the National Natural Sciences Foundation ofChina, No. 30671860
文摘AIM: To explore the inhibitory effects of pokeweed antiviral protein seed (PAP-S) and PAP encoded by a eukaryotic expression plasmid on hepatitis B virus (HBV) replication in vitro. METHODS: HepG2 2.2.15 cells in cultured medium were treated with different concentrations of PAP-S. HBsAg, HBeAg and HBV DNA in supernatants were determined by ELISA and fluorescent quantitative PCR respectively. MTT method was used to assay for cytotoxicity. HepG2 were cotransfected with various amounts of PAP encoded by a eukaryotic expression plasmid and replication competent wild-type HBV 1.3 fold over- length plasmid. On d 3 after transfection, HBsAg and HBeAg were determined by using ELISA. Levels of HBV core-associated DNA and RNA were detected by using Southern and Northern blot, respectively. RESULTS: The inhibitory effects of PAP-S on HBsAg, HBeAg and HBV DNA were gradually enhanced with the increase of PAP concentration. When the concentration of PAP-S was 10 μg/mL, the inhibition rates of HBsAg, HBeAg and HBV DNA were 20.9%, 30.2% and 50%, respectively. After transfection of 1.0 μg and 2.0 μg plasmid pXF3H-PAP, the levels of HBV nucleocapside- associated DNA were reduced by 38.0% and 74.0% respectively, the levels of HBsAg in the media by 76.8% and 99.7% respectively, and the levels of HBeAg by 72.7% and 99.3% respectively as compared with controls. Transfection with 2 μg plasmid pXF3H-PAP reduced the levels of HBV nucleocapside-associated RNA by 69.0%.CONCLUSION: Both PAP-S and PAP encoded by a eukaryotic expression plasmid could effectively inhibit HBV replication and antigen expression in vitro, and the inhibitory effects were dose-dependent.
基金The study was approved by the grants from the National Natural Science Foundation of China(No.81560730)the Training Project for Young and Middle-aged Chinese Medicine Leaders in Yunnan Province.
文摘To the Editor:Coronary heart disease(CHD)and depression are common in patients worldwide and have a high comorbidity rate.Researches on depression have shown that it has been linked with long-term risk for CHD.[1]The incidence and mortality of depression with CHD were higher,and the prognosis was worse.[2]The association between CHD and depression is well established and is suggested to be bidirectional.[3]Therefore,to provide possible conditions for the study of depression with CHD in the future,establishing a reliable animal model of depression with CHD is necessary.Chronic unpredictable mild stress(CUMS)is the most widely used to establish an animal model of depression for the past few years.Feeding a high-fat diet(HFD)to rats is a widely used method for establishing a model for atherosclerosis and CHD.Research reported that female cynomolgus monkeys that were housed in single cages and consumed an atherogenic diet could develop depression combined with CHD model.[4]Another paper reported that CUMS combined with HFD in rats was similar to the clinical manifestation of vascular depression in humans.[5]The former demonstrated that housing in single cages and feeding an atherogenic diet in monkeys could establish a depression combined with CHD model,the latter demonstrated that CUMS combined with HFD in rats was similar to vascular depression.It remains unclear whether CUMS combined with HFD in rats could establish a complex rat model of depression with CHD.Consequently,the aim of this study was to explore whether CUMS combined with HFD in rats could establish a representative,reliable rat model of depression with CHD and to provide possible conditions for an in-depth study of depression with CHD.
