Objective To study the protective effects of resveratrol against hepatic stellate cells (HSCs) and liver fibrogensis.Methods HSCs were isolated from liver of SD rats.The reactive oxygen output in HSCs under resveratrol in different concentrations was tested by DCFH-DA kit.The proliferation of HSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of HSCs was evaluated by Western blotting.The activity-related genes were measured by PCR.The models of liver fibrogenes were established.Resveratrol in different concentrations was administrated intraperitoneally.Liver was studied by pathology and SMA staining.Hydroxyproline content of liver and levels of collagen Ⅲ and hyaluronic acid in serum were tested.Results HSCs were isolated from liver and cultured successfully.Resveratrol inhibited the generation of the reactive oxygen.Proliferation and activation of HSCs was inhibited by resveratrol (0.536 ±0.052,0.411 ±0.047,0.327 ±0.063,0.312 ±0.032,F =12.776,P ＜0.05) (103 ±7,90 ±7,63 ± 4,53 ± 3,F =62.179,P ＜ 0.05).Resveratrol inhibited the expression of genes (myogenic determination gene MyoD,collagen 11 and collagen Ⅰ) in HSCs(122 ± 5,96 ± 3,68 ± 3,60 ± 3,F =180.600,P＜0.05) (100±8,82 ±3,53 ±3,51 ±2,F=77.451,P ＜0.05) (170 ±3,147 ±4,92 ±3,90 ±2,F =462.878,P ＜ 0.05).Resveratrol downregulated the level of hydroxyproline,collagen Ⅲ and hyaluronic acid (358.3 ± 20.2,320.5 ± 15.3,290.3 ± 24.5,F =23.929,P ＜ 0.05) (32.8 ± 3.1,28.9 ±1.3,25.3±1.8,F=20.050,P＜0.05)(276.3 ±17.8,225.3 ±28.3,195.4 ±11.2,F=18.585,P＜0.05).Conclusions Resveratrol can inhibit the proliferation and activation of HSCs and downregulate the fibrogensis level of the liver of rats.

Objective To investigate the Fsp27 gene's influence on the regulation of hepatic stellate cells (HSCs) in vitro.Methods HSCs were isolated from rat liver,the Fsp27 gene was detected in primary HSCs,and activated HSCs were detected by RT-qPCR.After 72 h of Fsp27 transduction through a lentivirus expressing Fsp27 (pLV-Fsp27),the proliferation of HSCs was tested by the CCK-8 test kit,smooth muscle α-actin (α-SMA) expression of HSCs was tested by Western blot,and the fibrosis-related genes were tested by RT-qPCR.Results The HSCs were isolated and cultured successfully,and the Fsp27 genetic difference between primary and activated HSCs was significant (P＜0.01).After coculture for 72 h,Fsp27 inhibited the proliferation and activation of HSCs (P＜0.05).Fsp27 can enhance expression of the MMP-2 gene and down-regulate expression of the TIMP-1 and TGF-β1 gene in activated HSCs (P＜0.05).Conclusion The Fsp27 gene can inhibit the proliferation and activation of HSCs,regulate the expression of fibrosis-related genes,and may play an important role in maintaining the quiescent phenotype of HSCs.

Objective To investigate the effect of capsaicin on hepatic stellate cells (HSCs) and liver fibrogenesis.Methods HSCs were cultured.The reactive oxygen in HSCs under capsaicin at different concentrations was tested by DCFH-DA kit.The proliferation of HSCs was detected by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of HSCs was evaluated by Western blot.The fibrosisrelated genes were tested by RT-PCR.The apoptosis of HSCs was measured by flow cytometer.Bcl-2,bax and cyt-c was detected by Western blot.A murine model of liver fibrogenes was established.Capsaicin of different concentration was injected intraperitoneally.Liver pathology was observed using HE staining.Hydroxyproline content of liver and levels of collagen Ⅲ and hyaluronic acid in serum were tested.Results In dose dependent manner capsaicin inhibited the generation of the reactive oxygen species.Proliferation and activation of HSCs was inhibited by capsaicin (respectively F =13.267,57.392,all P ＜ 0.05) and the apoptosis of HSCs was promoted by capsaicin (F =235.571,P ＜ 0.05).Bax,cyt-c and caspase-3 was increased obviously (respectively F =29.334,38.274,138.329,all P ＜ 0.05).Capsaicin changed the expression of fibrosis-related genes (TGF-β1,TIMP-1) in HSCs (respectively F =376.534,253.751,all P ＜0.05).Capsaicin downregulated the level of hydroxyproline,collagen Ⅲ and hyaluronic acid in the rat model (respectively F =153.397,27.149,38.392,all P ＜ 0.05).Conclusions Capsaicin inhibits the proliferation and activation of hepatic stellate cells.Capsaicin promotes the apoptosis of hepatic stellate cells,and inhibits liver fibrogenesis.

Objective To investigate the influence of fat-specific protein 27 (Fsp27) gene on the regulation of liver fibrogenesis in vivo.Methods Hepatic stellate cells (HSCs) were isolated from rat liver.Fsp27 gene was detected in primary HSCs and activated HSCs by real-time quantitative PCR (RTqPCR).Lentiviral vector carrying Fsp27 gene was constructed.The model of liver fibrosis was established by infusing carbon tetrachloride (CC14).The rats with liver fibrogenesis were infected by the virus.Liver sections were made to observe the structure and form of liver histocytes.The content of fibrous protein in liver and serum was detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay.Resukts HSCs were isolated and cultured successfully.The difference of Fsp27 gene between primary HSCs and activated HSCs was significant(P ＜ 0.01).The model of liver fibrosis was achieved.After infecting the model rats,we found the fibrosis level in treatment group was lower compared with control group.Conclusions Fsp27 treatment can decrease collagen deposition in the liver and inhibit the formation of fibrosis.

Objectives:To examine the effect of chloride blocker (NPPB and tamoxifen)on volume sensitive chloride channels in mouse cardiac ventricular myocytes. Methods:Isolated mouse cardiac ventricular myocytes were subjected to whole cell patch clamp to record the hypotonicity activated chloride currents. Results:When the myocytes were exposed to hypotonic solution, an obvious whole cell currents were activated. The currents were inhibited by extracellular NPPB reversibly and significantly. The specific blocker for volume sensitive chloride channel , tamoxifen (50 ?mol/L), could apparently block the activity of this channel in a voltage dependent manner. Conclusions:Mouse cardiac ventricular myocytes process volume sensitive chloride channel which is sensitive to NPPB and tamoxifen.

BACKGROUND:At present, liver transplantation is the best method to treat end-stage liver disease. UW solution is recognized as the best liver preservation solution, but it is expensive. Moreover, the extracel ular fluid of high K+condition is inconsistent with human physiology. Because transient hyperkalemia of UW solution often causes cardiac arrest, research and development of the new liver preservation solution already brook no delay. OBJECTIVE:To study the protective effect of self-designed KYL solution on ischemia reperfusion injury in macaque donor liver. METHODS:A total of eight recipient macaques and eight donor macaques were selected in this study. Each group contained KYL solution group (n=4) and UW solution group (n=4). Donor liver was perfused and cryopreserved for 4 hours and subjected to al ogenic orthotopic liver transplantation. At 30 minutes and 6 hours after transplantation, bile production was recorded. Blood was obtained and used to detect concentrations of aspartate aminotransferase, alanine aminotransferase, nitric oxide, endothelin-1 and tumor necrosis factor-α. Liver tissue was col ected and detected under the light microscope. RESULTS AND CONCLUSION:Bile secretion was found in both groups. Bile secretion production increased as time went on (P<0.05). At 30 minutes and 6 hours after donor liver reperfusion, serum aspartate aminotransferase and alanine aminotransferase concentrations were lower in the KYL solution group than in the UW solution group (P<0.05). No significant difference was found in levels of serum nitric oxide, endothelin 1 and tumor necrosis factor alpha between the two groups (P>0.05). Under light microscope, morphological observation of liver tissue revealed that cel ular edema was evident in the UW solution group than in the KYL solution group. Results suggest that the effect of KYL solution on preventing ischemia/reperfusion injury was identical to the UW solution, and partial effect was better than UW solution.

BACKGROUND:At present, liver transplantation is the only way to cure end-stage liver disease, but the complications after transplantation is stil an important factor of affecting the long-term survival of patients who received orthotopic liver transplantation, therefore it is necessary to establish a stable animal transplantation model. OBJECTIVE:To establish rat models of orthotopic liver transplantation. METHODS:After inhalation anesthesia with ether, 204 SD rats were perfused with 2-4℃ Ringer’s solution through the abdominal aorta. In order to reduce warm ischemia of the liver, the liver was not turned over before perfusion. The suprahepatic inferior vena cava was cut off along the phrenic ring after perfusion. No further trimming was needed when dressing, so as not to damage the vena cava. The donor liver was removed and preserved in 4℃Ringer’s liquid. The receptor liver was cut off and alogeneic orthotopic liver transplantation was performed using modified two-cuff method. After transplantation, rats could automaticaly turn over and drink water. Surviving more than 3 days is regarded as a successful transplantation. RESULTS AND CONCLUSION:102 liver transplantations were performed in 204 rats, with 86 rats surviving more than 3 days. The success rate of transplantation was 84%. The results demonstrate that rat models of orthotropic liver transplantation can be constructed successfuly through improving techniques.

BACKGROUND:Acelular dermal matrix is a cel-free natural tissue scaffold similar to human soft tissue, which is easy to shape and has non-toxic side effects. It has been used to repair the urethra and ureter. OBJECTIVE:To investigate the effect of acelular dermal matrix on the repair of bile duct injury. METHODS:Thirty Diannan miniature pigs were randomly divided into three groups: in blank group, the bile duct was resected folowed by end to end anastomosis; in experimental group, bile duct defect model was made folowed by repair with acelular dermal matrix; in control group, bile duct defect model was made folowed by repair with expanded polytetrafluoroethylene. At 6 and 24 weeks after repair, bile duct patches and surrounding tissues were taken for immunohistochemical observation and RT-PCR detection. RESULTS AND CONCLUSION: Compared with the control and blank group, the expression of cytokeratin was higher, but the expression of transforming growth factor β1 was lower in the experimental group. Within 24 weeks after repair, the total mRNA level of transforming growth factor β1 was lower in the experimental group than the other two groups (P < 0.05), but the total mRNA levels of insulin-like growth factor 2 and vascular endothelial growth factor were higher in the experimental group (P < 0.05). These findings indicate that the acelular dermal matrix for repair of bile duct injury can promote angiogenesis and bile duct epithelial regeneration, but not increase the formation of scars.

Objective To explore the relationship between angiogenin-1/2 (Ang-1/2) and clinical parameters of idiopathic pulmonary fibrosis (IPF), and to assess the value of Ang-1/2 in predicting the prognosis of patients with IPF.Methods A retrospective analysis was conducted. Ninety-one patients diagnosed as IPF by high resolution CT (HRCT) and lung biopsy admitted to Daqing Oil Field General Hospitalfrom March 2014 to January 2015 were enrolled. The general data, serum parameters and pulmonary function parameters of all patients were collected. After treatment, all of the 91 patients were followed-up to 2 years. The patients were divided into favorable prognosis group and unfavorable prognosis group according to follow-up results. The differences in all parameters between the two groups werecompared. The relationship between Ang-1, Ang-2 and lung function parameters was analyzed by Pearson correlation analysis. Cox proportional hazard regression model was used to evaluate the effect of clinical parameters on the prognosis of patients with IPF. The effect of Ang-2 in predicting prognosis of patients with IPF was analyzed by receiver operating characteristic (ROC) curve.Results During the 2-year follow-up period, 30 of 91 patients showed a favorable prognosis, and 55 showed an unfavorable prognosis with a poor prognosis rate of 64.71%, and 6 patients withdrew from the study due to loss of follow-up and death. Compared with the favorable prognosis group, Ang-2 level in the unfavorable prognosis group was significantly increased (μg/L: 2.88±1.63 vs. 1.89±1.22,t = 2.909,P= 0.005), but Ang-1 only showed a slight increase (μg/L: 28.70±14.26 vs. 25.62±11.95,t = 1.005,P = 0.318). The results of Pearson correlation analysis showed that Ang-2 level was negatively correlated with forced expiratory volume in 1 second (FVC1) and the percentage of carbon monoxide diffusing capacity accounting for the expected value (DLCO%;r value was -0.227 and -0.206, andP value was 0.147 and 0.253, respectively), but no significant correlation between the level of Ang-1 and FVC1 as well as DLCO% was found (r value was -0.153 and -0.121, andP value was 0.147 and 0.253, respectively). Cox proportional hazard regression model analysis showed that the prognosis of patients with IPF was significantly affected by smoking time and Ang-2 (bothP< 0.05), and the influence of Ang-2 was greater [relative risk (RR): 1.236 vs. 1.006, P= 0.037]. Age, gender, smoking and the levels of FVC1, DLCO% and Ang-1 had no significant effect on the prognosis of IPF patients (allP> 0.05). Prognostic analysis showed that the area under ROC curve (AUC) of Ang-2 for predicting prognosis of patients with IPF was 0.692, and the best diagnostic point was 0.35μg/L, the sensitivity was 61.8%, the specificity was 73.3%, the positive predictive value was 69.8%, and the negative predictive value was 65.7% which indicated that Ang-2 could predict the prognosis of patients with IPF.Conclusion Ang-2 could assess the prognosis of patients with IPF, which is expected to be used as an indicator of predicting the prognosis of patients with IPF.

Objective To evaluate the curative effect of selective decongestive devascularization shunt of gastrosplenic region(SDDS-GSR) for the treatment of portal hypertension. Methods From September 2000 to June 2008, 44 patients with portal hypertension had received SDDS-GSR in our hospital. Twenty-nine of them had been followed up for 12-85 months (mean=44months). Results Operative mortality was 0 %. Mesenteric area pressure(33.82±5.12 cm H_2O) was higher than splenic area pressure(24.57±4.63 cm H_2O)soon after the operation finished(P<0.01). No re-bleeding ca-ses were found, and the encephalopathy occurred in 2.27% of the patients in the early stage of post-operation. However, the rates of 3.45% for re-bleeding and 3.45% for encephalopathy were noticed in long-term follow-up. The 1-, 3- and 5-year survival were 100%, 95% and 95%, respectively. Dur-ing the long-term follow-up, the platelet counts markedly increased from (49.2±21.8 × 10~9/L) of preoperative value to (77.2±29.5×10~9/L) (P<0.01), while spleen size was significantly reduced.Conclusion SDDS-GSR is a reliable and reasonable surgical procedure for the management of portal hypertension.