AIM： To study the effect of oxymatrine-baicalin combination （OB） against HBV replication in 2.2.15 cells and α smooth muscle actin （ α SMA） expression, type I, collagen synthesis in HSC-T6 cells. METHODS： The ...AIM： To study the effect of oxymatrine-baicalin combination （OB） against HBV replication in 2.2.15 cells and α smooth muscle actin （ α SMA） expression, type I, collagen synthesis in HSC-T6 cells. METHODS： The 2.2.15 cells and HSC-T6 cells were cultured and treated respectively. HBsAg and HBeAg in the culture supernatants were detected by ELISA and HBV DNA levels were determined by fluorescence quantitative PCR. Total RNA was extracted from HSC-T6 cells and reverse transcribed into cDNA. The cDNAs were amplified by PCR and the quantities were expressed in proportion to β actin. The total cellular proteins extracted from HSC-T6 cells were separated by electrophoresis. Resolved proteins were electrophoretically transferred to nitrocellulose membrane. Protein bands were revealed and the quantities were corrected by β actin. RESULTS： In the 2.2.15 cell culture system, the inhibitory rate against secretion of HBsAg and HBeAg in the OB group was significantly stronger than that in the oxymatrine group （HBsAg, P = 0.043; HBeAg, P = 0.026; respectively）; HBV DNA level in the OB group was significantly lower than that in the oxymatrine group （P = 0.041）. In HSC-T6 cells the mRNA and protein expression levels of α SMA in the OB group were significantly lower as compared with those in the oxymatrine group （mRNA, P = 0.013; protein, P = 0.042; respectively）; The mRNA and protein expression levels of type I collagen in the OB group were significantly lower as compared with those in the oxymatrine group （mRNA, P 〈 0.01; protein, P 〈 0.01; respectively）.CONCLUSION： OB combination has a better effect against HBV replication in 2.2.15 cells and is more effective against α SMA expression and type I collagen synthesis in HSC-T6 cells than oxymatrine in vitro.展开更多
AIM： To investigate the role of emodin in protecting the liver against fibrogenesis caused by carbon tetrachloride （CCh） in rats and to further explore the underlying mechanisms. METHODS： Rat models of experimenta...AIM： To investigate the role of emodin in protecting the liver against fibrogenesis caused by carbon tetrachloride （CCh） in rats and to further explore the underlying mechanisms. METHODS： Rat models of experimental hepatic fibrosis were established by injection with CCh; the treated rats received emodin via oral administration at a dosage of 20 mg/kg twice a week at the same time. Rats injected with olive oil served as a normal group. Histopathological changes were observed by hematoxylin and eosin staining. The activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum and hepatic hydroxyproline content were assayed by biochemical analyses. The mRNA and protein relevant to hepatic stellate cell （HSC） activation in the liver were assessed using real-time reverse transcription-polymerase chain reaction （RT-PCR）, immunohistochernistry, western blotting and enzymelinked immunosorbent assay.RESULTS： The degree of hepatic fibrosis increased markedly in the CCh group compared to the normal group （P 〈 0.01）, and decreased markedly in the emodin group compared to the CCI4 group according to METAVIR scale （P 〈 0.01） compared with those in the normal control group （51.02 ± 10.64 IU/L and 132.28 ± 18.14 IU/L）. The activities of serum ALT and AST were significantly higher in rats injected with CCh （289.25 ± 68.84 IU/L and 423.89 ± 35.67 IU/L, both P 〈 0.05）. The activities of serum ALT and AST were significantly reduced by administration of emodin （176.34 ± 47.29 IU/L and 226.1 ± 44.52 IU/L, both P 〈 0.05）. Compared with the normal controls （54.53 ± 13.46 mg/g）, hepatic hydroxyproline content was significantly higher in rats injected with CCI4 （120.27 ± 28.47 mg/g, P 〈 0.05）. Hepatic hydroxyproline content was significantly reduced in the rats treated with emodin at 20 mg/kg （71.25 ± 17.02 mg/g, P 〈 0.05）. Emodin significantly protected the liver from injury by reducing serum AST and ALT activities and reducing hepa展开更多
Endocytosis occurs at the cell surface and involves internalization of the plasma membrane （PM） along with its constituent membrane proteins and lipids. Endocytosis is involved in sampling of the extracellular milie...Endocytosis occurs at the cell surface and involves internalization of the plasma membrane （PM） along with its constituent membrane proteins and lipids. Endocytosis is involved in sampling of the extracellular milieu and also serves to regulate various processes initiated at the cell surface. These include nutrient uptake, signaling from cell- surface receptors, and many other processes essential for cell and tissue functioning in metazoans. It is also central to the maintenance of PM lipid and protein homeostasis. There are multiple means of internalization that operate concurrently, at the cell surface. With advancement in high-resolution visualization techniques, it is now possible to track multiple endocytic cargo at the same time, revealing a remarkable diversity of endocytic processes in a single cell. A combination of live cell imaging and efficient genetic manipulations has also aided in understanding the functional hierarchy of molecular players in these mechanisms of internalization. Here we provide an account of various endocytic routes, their mechanisms of operation and occurrence across phyla.展开更多
Objective：To study the distribution of gelsolin in human platelet and plasma,and the association with blood-stasis syndrome（BSS） of coronary heart disease（CHD）.Methods：Sixty patients with CHD（30 in BSS group an...Objective：To study the distribution of gelsolin in human platelet and plasma,and the association with blood-stasis syndrome（BSS） of coronary heart disease（CHD）.Methods：Sixty patients with CHD（30 in BSS group and 30 in non-BSS group） and 30 healthy subjects（control group） were included in this study.The classification of the syndrome was based on clinical symptoms and signs.Gelsolin concentration in platelet rich plasma（PRP）,platelet poor plasma（PPP）,filamentous actin（F-actin） and group-specific component globulin （Gc-globulin） of PPP were determined by enzyme-linked immunosorbent assay（ELISA）.The fluorescence intensity of CD62p and cytoplasmic calcium（[Ca^（2＋）]_i） in human platelets of patients and healthy persons was measured with flow cytometry.Results：Compared with the control group,gelsolin in PRP of the BSS group increased significantly（P0.01）,while that in PPP of the BSS and non-BSS groups decreased markedly（P0.05）, the CD62p,[Ca^（2＋）]_i of platelet,F-actin,and Gc-globulin of the BSS and non-BSS groups increased significantly （P0.01）.Compared with the non-BSS group,the gelsolin concentration in PRP of BSS group increased significantly（P0.01）,the[Ca^（2＋）]_i of platelet of the BSS group increased markedly（P0.01）,while the F-actin and Gc-globulin of the BSS group had no statistical defference（P0.05）.Conclusions：Gelsolin concentration in PRP was increased and accompanied by the elevated[Ca^（2＋）]_i of platelet in CHD with BSS,while gelsolin in PPP were lowered markedly.We speculate that plasma gelsolin may clear F-actin from circulation,thus resulting in depletion of plasma gelsolin significantly.This,in addition to the increased calcium influx of platelets,may lead to the gelsolin abnormal expression on platelets during the process of BSS in CHD.Therefore,platelet gelsolin may serve as a new potential biomarker and a therapeutic target of BSS in CHD.展开更多
文摘AIM： To study the effect of oxymatrine-baicalin combination （OB） against HBV replication in 2.2.15 cells and α smooth muscle actin （ α SMA） expression, type I, collagen synthesis in HSC-T6 cells. METHODS： The 2.2.15 cells and HSC-T6 cells were cultured and treated respectively. HBsAg and HBeAg in the culture supernatants were detected by ELISA and HBV DNA levels were determined by fluorescence quantitative PCR. Total RNA was extracted from HSC-T6 cells and reverse transcribed into cDNA. The cDNAs were amplified by PCR and the quantities were expressed in proportion to β actin. The total cellular proteins extracted from HSC-T6 cells were separated by electrophoresis. Resolved proteins were electrophoretically transferred to nitrocellulose membrane. Protein bands were revealed and the quantities were corrected by β actin. RESULTS： In the 2.2.15 cell culture system, the inhibitory rate against secretion of HBsAg and HBeAg in the OB group was significantly stronger than that in the oxymatrine group （HBsAg, P = 0.043; HBeAg, P = 0.026; respectively）; HBV DNA level in the OB group was significantly lower than that in the oxymatrine group （P = 0.041）. In HSC-T6 cells the mRNA and protein expression levels of α SMA in the OB group were significantly lower as compared with those in the oxymatrine group （mRNA, P = 0.013; protein, P = 0.042; respectively）; The mRNA and protein expression levels of type I collagen in the OB group were significantly lower as compared with those in the oxymatrine group （mRNA, P 〈 0.01; protein, P 〈 0.01; respectively）.CONCLUSION： OB combination has a better effect against HBV replication in 2.2.15 cells and is more effective against α SMA expression and type I collagen synthesis in HSC-T6 cells than oxymatrine in vitro.
基金Supported by National Natural Science Foundation of China,No.30873396National Science Foundation for Post-doctoral Scientists of China,No.20080430140Qiqihar Foundation for Development of Science and Technology,China,No.05090
文摘AIM： To investigate the role of emodin in protecting the liver against fibrogenesis caused by carbon tetrachloride （CCh） in rats and to further explore the underlying mechanisms. METHODS： Rat models of experimental hepatic fibrosis were established by injection with CCh; the treated rats received emodin via oral administration at a dosage of 20 mg/kg twice a week at the same time. Rats injected with olive oil served as a normal group. Histopathological changes were observed by hematoxylin and eosin staining. The activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum and hepatic hydroxyproline content were assayed by biochemical analyses. The mRNA and protein relevant to hepatic stellate cell （HSC） activation in the liver were assessed using real-time reverse transcription-polymerase chain reaction （RT-PCR）, immunohistochernistry, western blotting and enzymelinked immunosorbent assay.RESULTS： The degree of hepatic fibrosis increased markedly in the CCh group compared to the normal group （P 〈 0.01）, and decreased markedly in the emodin group compared to the CCI4 group according to METAVIR scale （P 〈 0.01） compared with those in the normal control group （51.02 ± 10.64 IU/L and 132.28 ± 18.14 IU/L）. The activities of serum ALT and AST were significantly higher in rats injected with CCh （289.25 ± 68.84 IU/L and 423.89 ± 35.67 IU/L, both P 〈 0.05）. The activities of serum ALT and AST were significantly reduced by administration of emodin （176.34 ± 47.29 IU/L and 226.1 ± 44.52 IU/L, both P 〈 0.05）. Compared with the normal controls （54.53 ± 13.46 mg/g）, hepatic hydroxyproline content was significantly higher in rats injected with CCI4 （120.27 ± 28.47 mg/g, P 〈 0.05）. Hepatic hydroxyproline content was significantly reduced in the rats treated with emodin at 20 mg/kg （71.25 ± 17.02 mg/g, P 〈 0.05）. Emodin significantly protected the liver from injury by reducing serum AST and ALT activities and reducing hepa
文摘Endocytosis occurs at the cell surface and involves internalization of the plasma membrane （PM） along with its constituent membrane proteins and lipids. Endocytosis is involved in sampling of the extracellular milieu and also serves to regulate various processes initiated at the cell surface. These include nutrient uptake, signaling from cell- surface receptors, and many other processes essential for cell and tissue functioning in metazoans. It is also central to the maintenance of PM lipid and protein homeostasis. There are multiple means of internalization that operate concurrently, at the cell surface. With advancement in high-resolution visualization techniques, it is now possible to track multiple endocytic cargo at the same time, revealing a remarkable diversity of endocytic processes in a single cell. A combination of live cell imaging and efficient genetic manipulations has also aided in understanding the functional hierarchy of molecular players in these mechanisms of internalization. Here we provide an account of various endocytic routes, their mechanisms of operation and occurrence across phyla.
基金the National Natural Science Foundation of China (No.81073086)
文摘Objective：To study the distribution of gelsolin in human platelet and plasma,and the association with blood-stasis syndrome（BSS） of coronary heart disease（CHD）.Methods：Sixty patients with CHD（30 in BSS group and 30 in non-BSS group） and 30 healthy subjects（control group） were included in this study.The classification of the syndrome was based on clinical symptoms and signs.Gelsolin concentration in platelet rich plasma（PRP）,platelet poor plasma（PPP）,filamentous actin（F-actin） and group-specific component globulin （Gc-globulin） of PPP were determined by enzyme-linked immunosorbent assay（ELISA）.The fluorescence intensity of CD62p and cytoplasmic calcium（[Ca^（2＋）]_i） in human platelets of patients and healthy persons was measured with flow cytometry.Results：Compared with the control group,gelsolin in PRP of the BSS group increased significantly（P0.01）,while that in PPP of the BSS and non-BSS groups decreased markedly（P0.05）, the CD62p,[Ca^（2＋）]_i of platelet,F-actin,and Gc-globulin of the BSS and non-BSS groups increased significantly （P0.01）.Compared with the non-BSS group,the gelsolin concentration in PRP of BSS group increased significantly（P0.01）,the[Ca^（2＋）]_i of platelet of the BSS group increased markedly（P0.01）,while the F-actin and Gc-globulin of the BSS group had no statistical defference（P0.05）.Conclusions：Gelsolin concentration in PRP was increased and accompanied by the elevated[Ca^（2＋）]_i of platelet in CHD with BSS,while gelsolin in PPP were lowered markedly.We speculate that plasma gelsolin may clear F-actin from circulation,thus resulting in depletion of plasma gelsolin significantly.This,in addition to the increased calcium influx of platelets,may lead to the gelsolin abnormal expression on platelets during the process of BSS in CHD.Therefore,platelet gelsolin may serve as a new potential biomarker and a therapeutic target of BSS in CHD.
