Previously, we have purified Jerdonitin from Trimeresurusjerdonii venom. Compared with other P-Ⅱ class snake venom metalloproteinases （SVMPs）, Jerdonitin has a primary structure comprising metalloproteinase and dis...Previously, we have purified Jerdonitin from Trimeresurusjerdonii venom. Compared with other P-Ⅱ class snake venom metalloproteinases （SVMPs）, Jerdonitin has a primary structure comprising metalloproteinase and disintegrin domains. However, no hemorrhagic and fibrinogenolytic activities were detected for Jerdonitin. We thought that organic buffer of high performance liquid charamatography （HPLC） might affect its enzymatic activity. In this study, we purified Jerdonitin by another procedure excluding the HPLC. It was homogenous as judged by SDS-PAGE and had an apparent molecular weight of 36 kDa under non-reducing conditions and 38 kDa under reducing conditions, respectively. Like other typical metalloproteinases, Jerdonitin preferentially degraded alpha-chain of human fibrinogen and this fibrinogenolytic activity was completely inhibited by EDTA, but not by PMSF. It was interesting that Jerdonitin did not induce hemorrhage after intradermal injection in mice.展开更多
Objective: To determine the effects of albumin administration on lung injury and apoptosis in traumatic/hemorrhagic shock (T/HS) rats. Methods: Studies were performed on an in vivo model of spontaneously breathing rat...Objective: To determine the effects of albumin administration on lung injury and apoptosis in traumatic/hemorrhagic shock (T/HS) rats. Methods: Studies were performed on an in vivo model of spontaneously breathing rats with induced T/HS; the rats were subjected to femur fracture, ischemia for 30 min, and reperfusion for 20 min with Ringer's lactate solution (RS) or 5% (w/v) albumin (ALB), and the left lower lobes of the lungs were resected. Results: Albumin administered during reperfusion markedly attenuated injury of the lung and decreased the concentration of lactic acid and the number of in situ TdT-mediated dUTP nick-end labelling (TUNEL)-positive cells. Moreover, immunohistochemistry performed 24 h after reperfusion revealed increases in the level of nuclear factor κB (NF-κB), and phosphorylated p38 mitogen-activated protein kinase (MAPK) in the albumin-untreated group was down-regulated by albumin treatment when compared with the sham rats. Conclusion: Resuscitation with albumin attenuates tissue injury and inhibits T/HS-induced apoptosis in the lung via the p38 MAPK signal transduction pathway that functions to stimulate the activation of NF-κB.展开更多
Objective To explore the potential mechanism of breakthrough bleeding associated with progestin with in vitro methods.Methods The isolation and culture of human endometrial endothelial cells(HEECs)was performed with t...Objective To explore the potential mechanism of breakthrough bleeding associated with progestin with in vitro methods.Methods The isolation and culture of human endometrial endothelial cells(HEECs)was performed with the method established in our laboratory.The content and activity of urokinase-type plasminogen activator(uPA)and the content of plasminogen activator inhibitor-1(PAI-1)in cell supernatants after incubated with different concentrations of progesterone(0-5 μmol/L)and 17β-estradiol(0,0.1,or 1 nmol/L)were measured by method of ELISA.Apoptosis rate of HEECs was measured by flow cytometry.Viable cell count was measured by MTT.Results The increased level of progesterone(0.5-5 μmol/L)combined with 17β-estradiol elevated content and activity of uPA while the production of PAI-1 remained unchanged.The apoptosis of HEECs was inhibited along with the increment of total viable cell counts at higher concentrations of progesterone with 17β-estradiol.Conclusion The inhibition of apoptosis and increased content and activity of uPA may contribute to the occurrence of irregular bleeding associated with progestin use to some extent.展开更多
文摘Previously, we have purified Jerdonitin from Trimeresurusjerdonii venom. Compared with other P-Ⅱ class snake venom metalloproteinases （SVMPs）, Jerdonitin has a primary structure comprising metalloproteinase and disintegrin domains. However, no hemorrhagic and fibrinogenolytic activities were detected for Jerdonitin. We thought that organic buffer of high performance liquid charamatography （HPLC） might affect its enzymatic activity. In this study, we purified Jerdonitin by another procedure excluding the HPLC. It was homogenous as judged by SDS-PAGE and had an apparent molecular weight of 36 kDa under non-reducing conditions and 38 kDa under reducing conditions, respectively. Like other typical metalloproteinases, Jerdonitin preferentially degraded alpha-chain of human fibrinogen and this fibrinogenolytic activity was completely inhibited by EDTA, but not by PMSF. It was interesting that Jerdonitin did not induce hemorrhage after intradermal injection in mice.
基金Project supported by the National Natural Science Foundation of China (No. 30672071) the Traditional Chinese Medicine Foun- dation of Zhejiang Province, China (No. 2004C071)
文摘Objective: To determine the effects of albumin administration on lung injury and apoptosis in traumatic/hemorrhagic shock (T/HS) rats. Methods: Studies were performed on an in vivo model of spontaneously breathing rats with induced T/HS; the rats were subjected to femur fracture, ischemia for 30 min, and reperfusion for 20 min with Ringer's lactate solution (RS) or 5% (w/v) albumin (ALB), and the left lower lobes of the lungs were resected. Results: Albumin administered during reperfusion markedly attenuated injury of the lung and decreased the concentration of lactic acid and the number of in situ TdT-mediated dUTP nick-end labelling (TUNEL)-positive cells. Moreover, immunohistochemistry performed 24 h after reperfusion revealed increases in the level of nuclear factor κB (NF-κB), and phosphorylated p38 mitogen-activated protein kinase (MAPK) in the albumin-untreated group was down-regulated by albumin treatment when compared with the sham rats. Conclusion: Resuscitation with albumin attenuates tissue injury and inhibits T/HS-induced apoptosis in the lung via the p38 MAPK signal transduction pathway that functions to stimulate the activation of NF-κB.
文摘Objective To explore the potential mechanism of breakthrough bleeding associated with progestin with in vitro methods.Methods The isolation and culture of human endometrial endothelial cells(HEECs)was performed with the method established in our laboratory.The content and activity of urokinase-type plasminogen activator(uPA)and the content of plasminogen activator inhibitor-1(PAI-1)in cell supernatants after incubated with different concentrations of progesterone(0-5 μmol/L)and 17β-estradiol(0,0.1,or 1 nmol/L)were measured by method of ELISA.Apoptosis rate of HEECs was measured by flow cytometry.Viable cell count was measured by MTT.Results The increased level of progesterone(0.5-5 μmol/L)combined with 17β-estradiol elevated content and activity of uPA while the production of PAI-1 remained unchanged.The apoptosis of HEECs was inhibited along with the increment of total viable cell counts at higher concentrations of progesterone with 17β-estradiol.Conclusion The inhibition of apoptosis and increased content and activity of uPA may contribute to the occurrence of irregular bleeding associated with progestin use to some extent.
