AIM:To observe the effect of vincristine on hepatitis B virus(HBV) replication in vitro and to study its possible mechanisms.METHODS:Vincristine was added to the cultures of two cell lines stably expressing HBV.Then,t...AIM:To observe the effect of vincristine on hepatitis B virus(HBV) replication in vitro and to study its possible mechanisms.METHODS:Vincristine was added to the cultures of two cell lines stably expressing HBV.Then,the levels of hepatitis B surface antigen(HBs Ag),hepatitis B e antigen(HBe Ag),and hepatitis B core antigen(HBc Ag) in the supernatants or cytoplasm were examined using by enzyme-linked immunosorbent assay and Western blot.The HBV pregenome RNA(pg RNA) was detected using reverse transcription-PCR and realtime fluorescent quantitative PCR(RT-q PCR),and viral DNA was detected using Southern blot and RT-q PCR.Cell proliferation after drug treatment was detected using the Brd U incorporation test and the trypan blue exclusion assay.Cell cycle and cell apoptosis were examined using flow cytometry and Western blot.RESULTS:Vincristine up-regulated HBV replication directly in vitro in a dose-dependent manner,and 24-h exposure to 0.1 μmol/L vincristine induced more than 4-fold and 3-fold increases in intracellular HBV DNA and the secretion of viral DNA,respectively.The expression of HBV pg RNA,intracellular HBs Ag and HBc Ag,and the secretion of HBe Ag were also increased significantly after drug treatment.Most importantly,vincristine promoted the cell excretion of HBV nucleocapsids instead of HBV Dane particles,and the nucleocapsids are closely related to the HBV pathogenesis.Furthermore,vincristine inhibited the proliferation of cells stably expressing HBV.The higher the concentration of the drug,the more significant the inhibition of the cell proliferation and the stronger the HBV replication ability in cells.Flow cytometry indicated that cell cycle arrest at S-phase was responsible for the cell proliferation inhibition.CONCLUSION:Vincristine has a strong stimulatory effect on HBV replication and induces cell cycle arrest,and cell proliferation inhibition may be conducive to viral replication.展开更多
In this paper, we developed a novel process integrating vacuum distillation with atmospheric chlorination reaction(VD-ACR) to realize the flexible production of tetrachloroethane(TeCA) and pentachloroethane(PCA)from 1...In this paper, we developed a novel process integrating vacuum distillation with atmospheric chlorination reaction(VD-ACR) to realize the flexible production of tetrachloroethane(TeCA) and pentachloroethane(PCA)from 1,2-dichloroethane(DCA). During the simulation, the distillation column and reactors were operated for separation and chlorination respectively under variable pressures and temperatures. It is interesting to note that VD-ACR processes producing pure TeCA or PCA can exhibit the similar configuration parameters after optimization, which enables the flexible production of TeCA and PCA with different molar ratios via changing operating parameters. The molar ratio of TeC A/PCA can be fine-tuned within the range of 0.9:0.1-0.1:0.9 through adjusting the amount of chlorine pumped into side reactors, giving rise to the increase of the heat duty of reboiler by five times. A pilot-scale experiment was then operated based-upon this VD-ACR process and the result matched well with the simulation. Therefore, the VD-ACR model presented in this study will be beneficial for the industrial-scale flexible production of TeCA and PCA from DCA.展开更多
基金Supported by National Natural Science Foundation of China,No.81201282 and No.30901280Grants from Chongqing Natural Science Foundation,No.cstc2012jj A10047the PhD Programs Foundation of the Ministry of Education of China,No.20125503120004
文摘AIM:To observe the effect of vincristine on hepatitis B virus(HBV) replication in vitro and to study its possible mechanisms.METHODS:Vincristine was added to the cultures of two cell lines stably expressing HBV.Then,the levels of hepatitis B surface antigen(HBs Ag),hepatitis B e antigen(HBe Ag),and hepatitis B core antigen(HBc Ag) in the supernatants or cytoplasm were examined using by enzyme-linked immunosorbent assay and Western blot.The HBV pregenome RNA(pg RNA) was detected using reverse transcription-PCR and realtime fluorescent quantitative PCR(RT-q PCR),and viral DNA was detected using Southern blot and RT-q PCR.Cell proliferation after drug treatment was detected using the Brd U incorporation test and the trypan blue exclusion assay.Cell cycle and cell apoptosis were examined using flow cytometry and Western blot.RESULTS:Vincristine up-regulated HBV replication directly in vitro in a dose-dependent manner,and 24-h exposure to 0.1 μmol/L vincristine induced more than 4-fold and 3-fold increases in intracellular HBV DNA and the secretion of viral DNA,respectively.The expression of HBV pg RNA,intracellular HBs Ag and HBc Ag,and the secretion of HBe Ag were also increased significantly after drug treatment.Most importantly,vincristine promoted the cell excretion of HBV nucleocapsids instead of HBV Dane particles,and the nucleocapsids are closely related to the HBV pathogenesis.Furthermore,vincristine inhibited the proliferation of cells stably expressing HBV.The higher the concentration of the drug,the more significant the inhibition of the cell proliferation and the stronger the HBV replication ability in cells.Flow cytometry indicated that cell cycle arrest at S-phase was responsible for the cell proliferation inhibition.CONCLUSION:Vincristine has a strong stimulatory effect on HBV replication and induces cell cycle arrest,and cell proliferation inhibition may be conducive to viral replication.
基金Supported by the National Natural Science Foundation of China(21276126,61203020)Prospective Joint Research Project of Jiangsu Province(BY2014005-02,BY2015005-02)The Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
文摘In this paper, we developed a novel process integrating vacuum distillation with atmospheric chlorination reaction(VD-ACR) to realize the flexible production of tetrachloroethane(TeCA) and pentachloroethane(PCA)from 1,2-dichloroethane(DCA). During the simulation, the distillation column and reactors were operated for separation and chlorination respectively under variable pressures and temperatures. It is interesting to note that VD-ACR processes producing pure TeCA or PCA can exhibit the similar configuration parameters after optimization, which enables the flexible production of TeCA and PCA with different molar ratios via changing operating parameters. The molar ratio of TeC A/PCA can be fine-tuned within the range of 0.9:0.1-0.1:0.9 through adjusting the amount of chlorine pumped into side reactors, giving rise to the increase of the heat duty of reboiler by five times. A pilot-scale experiment was then operated based-upon this VD-ACR process and the result matched well with the simulation. Therefore, the VD-ACR model presented in this study will be beneficial for the industrial-scale flexible production of TeCA and PCA from DCA.
