Q Sepharose Fast Flow(Q Sepharose F.F.) was adopted to purify the suspensions of type Ⅰ,Ⅱ,Ⅲ oral poliovaccine (OPV), which is prepared from Vero cells. After clarification and concentration by ultrafiltration,th...Q Sepharose Fast Flow(Q Sepharose F.F.) was adopted to purify the suspensions of type Ⅰ,Ⅱ,Ⅲ oral poliovaccine (OPV), which is prepared from Vero cells. After clarification and concentration by ultrafiltration,the recovery of virus infected titre may attain above 85%.Using column chromatography on Q Sepharose F.F. to purify the concentrated virus suspensions,the purified viruses attain 100% recovery of virus infectivity. Dot membrane hybrization was used to detect DNA with the probe of Vero cell genome DNA which was labeled with α P 32 dATP,the contents of residual substrate DNA was less than 100 Pg/dose. The process of downstream had no significant influence on some biological characters of purified viruses,such as virus morphology ,tumorigenicity, rct/40 character and capsid protein.This downstream reseach indicates that Q Sepharose F.F. is an ideal chromatography material for purifying OPV prepared from Vero cells.展开更多
Dexamethasone—a potent synthetic glucocorticoid—has multiple diagnostic and therapeutic applications in wide range of age groups. However, the side-effects of dexamethasone (Dex) treatment including those on develop...Dexamethasone—a potent synthetic glucocorticoid—has multiple diagnostic and therapeutic applications in wide range of age groups. However, the side-effects of dexamethasone (Dex) treatment including those on development are becoming increasingly apparent. Since the developmental processes are energy-dependent, we examined the effects of chronic Dex treatment on kinetics properties of liver mitochondrial F0.F1-ATPase and mitochondrial membrane lipid profiles in rats belonging to different developmental age groups (2, 3, 4 and 5 weeks) and in adults (~8 weeks). The animals were treated with a subcutaneous dose of 2 mg of Dex/kg body weight (or saline as vehicle) for three alternative days (at around 7.00 A.M.) prior to the day of sacrifice. Dex treatment resulted in significant reduction in F0.F1-ATPase activity in developmental age groups and in adults as compared to their age-matched vehicle-treated control group. The substrate kinetics analysis of F0.F1-ATPase resolved Km and Vmax values in 3 components in all the control age groups;whereas Dex treatment significantly altered the Km and Vmax values or abolished the entire components in age-specific manner. Dex treatment significantly lowered the energy of activation and altered phase transition temperature (TtoC) in all the developmental age groups and in adults. Dex treatment significantly increased the contents of total phospholipid (TPL), individual phospholipids classes and cholesterol (CHL) in all the developmental age groups whereas opposite pattern was observed in adults. The mitochondrial membrane became more fluidized in the developing age groups (2, 4 and 5 weeks);whereas no change was observed in 3-week and adult groups following Dex treatment. In present study, our data demonstrate comprehensive deleterious effects of chronic Dex treatment on liver mitochondrial membrane structure and F0.F1-ATPase functional properties with respect to energy metabolism. At the same time, our data also warns against excessive repeated use of antenatal DEX in treatments in growing and adult human patients.展开更多
文摘Q Sepharose Fast Flow(Q Sepharose F.F.) was adopted to purify the suspensions of type Ⅰ,Ⅱ,Ⅲ oral poliovaccine (OPV), which is prepared from Vero cells. After clarification and concentration by ultrafiltration,the recovery of virus infected titre may attain above 85%.Using column chromatography on Q Sepharose F.F. to purify the concentrated virus suspensions,the purified viruses attain 100% recovery of virus infectivity. Dot membrane hybrization was used to detect DNA with the probe of Vero cell genome DNA which was labeled with α P 32 dATP,the contents of residual substrate DNA was less than 100 Pg/dose. The process of downstream had no significant influence on some biological characters of purified viruses,such as virus morphology ,tumorigenicity, rct/40 character and capsid protein.This downstream reseach indicates that Q Sepharose F.F. is an ideal chromatography material for purifying OPV prepared from Vero cells.
文摘Dexamethasone—a potent synthetic glucocorticoid—has multiple diagnostic and therapeutic applications in wide range of age groups. However, the side-effects of dexamethasone (Dex) treatment including those on development are becoming increasingly apparent. Since the developmental processes are energy-dependent, we examined the effects of chronic Dex treatment on kinetics properties of liver mitochondrial F0.F1-ATPase and mitochondrial membrane lipid profiles in rats belonging to different developmental age groups (2, 3, 4 and 5 weeks) and in adults (~8 weeks). The animals were treated with a subcutaneous dose of 2 mg of Dex/kg body weight (or saline as vehicle) for three alternative days (at around 7.00 A.M.) prior to the day of sacrifice. Dex treatment resulted in significant reduction in F0.F1-ATPase activity in developmental age groups and in adults as compared to their age-matched vehicle-treated control group. The substrate kinetics analysis of F0.F1-ATPase resolved Km and Vmax values in 3 components in all the control age groups;whereas Dex treatment significantly altered the Km and Vmax values or abolished the entire components in age-specific manner. Dex treatment significantly lowered the energy of activation and altered phase transition temperature (TtoC) in all the developmental age groups and in adults. Dex treatment significantly increased the contents of total phospholipid (TPL), individual phospholipids classes and cholesterol (CHL) in all the developmental age groups whereas opposite pattern was observed in adults. The mitochondrial membrane became more fluidized in the developing age groups (2, 4 and 5 weeks);whereas no change was observed in 3-week and adult groups following Dex treatment. In present study, our data demonstrate comprehensive deleterious effects of chronic Dex treatment on liver mitochondrial membrane structure and F0.F1-ATPase functional properties with respect to energy metabolism. At the same time, our data also warns against excessive repeated use of antenatal DEX in treatments in growing and adult human patients.
