Objective To observe the expression changes of cytokine RhoA,stress fiber and Vinculin in the formation of hepatic fibrosis in rats, and investigate the role of RhoA in hepatic fibrosis. Methods Forty male SD rats were divided into two groups:model group and control group. The rats in the model group were given 60% CCl 4 by subcutaneous injection(1 ml/kg). The liver tissue was taken out for research at 1st、4th and 8th week respectively, RhoA and Vinculin mRNAs were detected by RT PCR, Vinculin and F actin proteins were detected by immunohistochemistry and immunofluorescence respectively, RhoA and Vinculin proteins were detected by Western blotting. Results Compared with control group, the expressions of mRNA of RhoA and Vinculin, the proteins of Vinculin, F actin and RhoA were all significantly increased( P

Objective To study the development a clinical useful microarray which can detect 4 most commonly and well documented sites for detection of lamivudine-resistance mutants in HBV lamivudine therapy. Methods (1) The selected 16 oligonucleotide probes to screen 4 mutated sites in YMDD region are immoblized in a specific treatment slice by GMS 417 Arrayer. The target 308 bp nucleic acids segment of HBV were amplificated and labeled with Cy5-dCTP fluorescent dye by PCR. After hybridization of target DNA with microarray, the microchips were scanned with Gene TAC LS IV scanner and the data were obtained after processed in computer. (2) To evaluated the specialty and reproducibility of the microarray, the plasmid with Leu515Met, Met539Ile, Met539Val,V542I mutations were condstructed and serve as positive sample. Another 50 sera of lamivudine treated Hepatitis B patients for 6 to 12 months as well as 50 sera without lamivudine administration were assayed by this microarray to evaluated the rate and genetype in lamivudine related resistance mutation. Results The microarray can identify a single base change of selected lamivudine resistance-related mutation and multiple mutation detection by a single assay as well. The specialty are well in agreement with sequencing for 98%. The reproducibility rate of repeat examination is range from 96%-100%. Conclusion The microarray of lamivuine resistance related mutation can identify a single base change as multiple mutations well. And its hight reproducible results may be useful for clinical monitoring lamivudine resistance related HBV mutation.

The correlation between pulmonary endothelin receptors and alveolar-arterial oxygen gradient (A-aDO2) in rats with hepatopulmonary syndrome was investigated. Animals were divided into 2 groups: Sham-operated (Sham) group and common bile duct ligation (CBDL) group. Arterial blood gas was evaluated by a blood gas analyzer. The concentrations of ET-1 in blood and lung tissue sample were evaluated by radioimmunoassay. The distribution and expression of two kinds of subtype receptor of ET-1, ETRA and ETRB were examined by in situ hybridization. The results showed that the level of A-aDO2 was higher in CBDL group than that in Sham group (P < 0.05). The levels of plasma and pulmonary ET-1 in CBDL group were both higher than in Sham group (P < 0.05). There was no significant difference in average A of ETRA between two groups by imaging analysis (0.21 +/- 0.06 vs 0.22 +/- 0.08, P > 0.05), while that of ETRB was higher in CBDL group than in Sham group (0.58 +/- 0.16 vs 0.28 +/- 0.07, P < 0.05). The expression of ETRB in lung was positively correlated with A-aDO2 (P < 0.05). It was concluded that the widened A-aDO2 may be related with enhancement of the expression of ETRB in lung.
Endothelin-1/metabolism ; Hepatopulmonary Syndrome/*metabolism ; Lung/*metabolism ; Oxygen/*metabolism ; Pulmonary Alveoli/*metabolism ; Rats, Wistar ; Receptor, Endothelin A/metabolism ; Receptor, Endothelin B/*metabolism

AIM: To investigate the different vasoactive effects of nitric oxide (NO) and endothelin (ET) on splanchnic arterial and venous vessels in cirrhotic rats. METHODS: Cirrhosis was induced in Wistar rats by subcutaneously administration of carbon tetrachloride. Maximal relaxation (Rmax) and contraction (Cmax) to NO and ET were determined in vitro using isolated vascular strips prepared from portal vein (PV) and mesenteric artery (MA) of both cirrhotic and normal rats, and EC50 was calculated for effects of NO and ET, respectively. RESULTS: Rmax of PV and MA to sodium nitroprusside (SNP) (releasing NO) were significantly higher in cirrhotic rats (n=8) than those in normal rats (n=7), and EC50 of NO were dramatically lower in cirrhotic rats than those in control (P

The correlation between pulmonary endothelin receptors and alveolar-arterial oxygen gradient (A-aDO2) in rats with hepatopulmonary syndrome was investigated. Animals were divided into 2 groups: Sham-operated (Sham) group and common bile duct ligation (CBDL) group. Arterial blood gas was evaluated by a blood gas analyzer. The concentrations of ET-1 in blood and lung tissue sample were evaluated by radioimmunoassay. The distribution and expression of two kinds of subtype receptor of ET-1, ETRA and ETRB were examined by in situ hybridization. The results showed that the level of A-aDO2 was higher in CBDL group than that in Sham group (P＜0.05).The levels of plasma and pulmonary ET-1 in CBDL group were both higher than in Sham group (P ＜0.05). There was no significant difference in average A of ETRA between two groups by imaging analysis (0.21±0.06 vs 0.22±0.08, P＞0.05), while that of ETRB was higher in CBDL group than in Sham group (0. 58±0.16 vs 0.28±0.07, P＜0.05). The expression of ETRB in lung was positively correlated with A-aDO2 (P＜0.05). It was concluded that the widened A-aDO2 may be related with enhancement of the expression of ETRB in lung.

Animals ; Cell Adhesion Molecules ; analysis ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; analysis ; L-Selectin ; analysis ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis

Animals ; Blood Pressure ; physiology ; Hemodynamics ; physiology ; Hypertension, Portal ; blood ; metabolism ; physiopathology ; Liver Cirrhosis, Experimental ; blood ; metabolism ; physiopathology ; Male ; Neuropeptide Y ; blood ; metabolism ; Portal Pressure ; physiology ; Rats ; Rats, Sprague-Dawley ; Sodium ; urine