Objective To investigate the joint effects of selective digestive decontamination (SDD) and glutamine (Gln) on preventing intestinal bacterial translocation of orthotopic piggyback liver transplantation and to observe the incidence of postoperative pneumonia in rabbit. Methods Thirty rabbits received orthotopic piggyback liver transplantation and were randomly divided into three groups (SDD group, SDD+Gln group and control group). Mixed emulsion of tobramycin, polymyxin E and nystatin were given to the rabbits in SDD group. Same dosage of the above components plus Gln were given to the rabbits in SDD+Gln group. Samples of portal vein blood, ileum tissue and lung tissue were obtained in each group at different phases during and after operation, the pathological changes of ileum tissue, the bacterial translocation in blood of portal vein and the incidence of postoperative pneumonia were detected. Results The mixing section area of intestinal blood capillaries in SDD+Gln group was smaller compared with control group (P

Objective To investigate the effect of CaMK Ⅱ expression on apoptosis of rat hepatocytes BRL-3A.Methods Rat BRL-3A cells were stable passage were cultured.The CaMK Ⅱ γ protein (LV-CaMK Ⅱ γ group) and CaMK Ⅱ γshRNA (shRNA group) lentiviral expression systems were constructed.The corresponding blank vectors (LV-NC group and shRNA-NC group) and normal saline (CON group) were perfused into the control groups.The expression levels of CaMK Ⅱ,Cyt C and MF proteins were detected by Western blotting,and the apoptosis rate of BRL-3A cells was measured by Tunel method.Results The protein expression of CaMK Ⅱ,Cyt C and AIF in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).The protein expression of CaMK Ⅱ,Cyt C and AIF in shRNA group was significantly lower than that in CON group (P＜ 0.05).There was no significant difference among CON group,LV-NC group and shRNA-NC group (P＞0.05).At the same time point,the apoptosis rate of hepatocytes in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).At the same time point,the apoptosis rate of hepatocytes in shRNA group was significantly higher than in CON group (P＜0.05).There was no significant difference in the apoptosis of hepatocytes among CON group,LV-NC group and shRNA-NC group (P＞0.05).Conclusion The specific CaMK Ⅱ signaling pathway can inhibit the apoptosis of BRL-3A cells,while the enhanced CaMK Ⅱ signaling pathway promotes the apoptosis of BRL-3A cells.

Objective To study the mechanism of immune hyporesponsiveness of allograft rejection induced by transfusion nonpufsed allopeptide syngeneic immature dendritic cell(imDC) generated from recipient bone marrow progenitors and to explore a possible strategy for liver allograft protection in clinic.Methods Forty experimental rats were randomly divided into 4 group: control group,cyclosporine A(CsA) group,mature DC(mDC) group and imDC group.In control group,Wistar rats only received liver transplantation.In CsA group,Wistar rats underwent liver transplantation plus CsA treatment(10 mg/(kg?d)).In mDC group,recipient-derived mDC 1?106 were infused intravenously through the penile vein to Wistar rats.In imDC group,ImDC with the dose of 1?106 were injected into Wistar rats via the dorsum vein of penile.In each group,five recipients were killed on the 10th day after transplantation,the other five recipients were left to observe survival time.The levels of ALT,AST,TBIL,IL-2,IFN-?,IL-4 and IL-10 were detected.The acute rejection and the expression of FasL/Fas in the grafts were detected by HE and immunohistochemical staining.Western blot was used to detect Scurfin protein expression of CD4+ CD25+ T cells.Results The median survival time of the liver allografts in CsA group and imDC group were significantly longer than that in control group and mDC group(P

Objective To explore improvement of orthotopic liver transplantation model in rhesus monkey.Methods Healthy rhesus monkeys were chosen to perform orthotopic liver transplantation for 10 cases.The model was established by drawing on a variety of animal model methods,and the portal vein cuff method was used to establish stable model of orthotopic liver transplantation in rhesus monkeys.Results Ten orthotopic liver transplantation models in rhesus were performed,and the achievement ratio of operation was 10/10.The time of donor hepatectomy and donor preparation was (20?5) min and (30?7) min,respectively.The operation time of recipient and anhepatic phase were (180?35) min and (17?4) min,respectively.After 24 h of operation 9 cases survived,one case died of intra-abdominal hemorrhage after 9 h of operation.After 72 h of operation 8 cases survived,and one case died of upper gastrointestinal bleeding after 38 h of operation.After one week of operation 5 cases survived,and 3 cases died of rejection after 9,11,and 11 d of operation,respectively.The longest survival time was 32 d,but all of them also died of rejection.No portal vein thrombosis and biliary complications were found in all recipients.Conclusion The improved rhesus monkey model of orthotopic liver transplantation is easy to perform with high achievement ratio of operation.It is an ideal animal model for pre-clinical studies of liver transplantation.

Objective To improve some surgical model techniques of rat orthotopic liver transplantation.Methods Two-cuff(portal vein and infrahepatic vena cava) technique was applied to the production of the animal model in SD and Wistar rats.Results With the improvements a successful rate of 85% was achieved in the production of the experimental model with anhepatic period of an average 21 minutes.Conclusion The results show that the model is stable and can be used in the experiment of liver transplantation in the rats.

Objective To investigate the effect of recombinant human growth hormone(rhGH)on the protein expression of NF-?B P65 in ischemic reperfusion injury of rat liver.Methods One hundred male rats models were constructed by Pringle's method and were randomly divided into two groups: the rhGH group and the control group.In rhGH group, the rats were injected(0.2IU/100g weight)seven days before the ischemic reperfusion injury,while in control group,rats was replaced by normal sodium.The levels of protein expression of NF-?B P65 were detected by Western blotting and immunohistochemistry.Results Compared with control group,the expression of NF-?B P65 and NF-?B activity(P

BACKGROUND:The proteome is a highlight technology in medical research fields lately, and has been reported to be applied in basic research fields related to liver transplantation. However, it has not been heard that the proteome has been used in research related to reduced-size liver transplantation. OBJECTIVE: To study expression of hepatic differential proteins related to signal transduction using proteomics after reduced-size liver transplantation in rats. METHODS:On the basis of successful establishment of rat models of reduced-size liver transplantation, transplanted liver tissues were obtained at 1, 3 and 7 days after transplantation. Postoperative liver tissue and normal donor, receptor liver tissues were subjected to solid pH gradient two-dimensional gel electrophoresis. Two-dimensional gel electrophoresis patterns were set up. Differentialy expressed protein spots were identified using tandem mass spectrometry analysis and database. RESULTS AND CONCLUSION:Seventy-two differential protein stains were found taking 10 times measure. Finaly, 32 proteins with clear functions were identified. Of them, four proteins participated in signal transduction, and they distributed at 3 and 7 days after liver transplantation, accounting for 6%. Results verified that on the basis of successful and stable establishment of rat models of reduced-size liver transplantation, proteomics technology was utilized to study differential proteins involving in signal transduction after reduced-size liver transplantation, and this study provides data for further deep investigation of regulating MicroRNA of these proteins.

Abstract BACKGROUND: Looking for the early diagnosis of acute rejection indicators after liver transplantation can assess the risk after liver transplantation quickly and effectively, and T lymphocytes play the significant role in acute rejection. OBJECTIVE:To observe the relationship between acute rejection and variation of expression of T cel subset in blood after liver transplantation in rhesus monkey. METHODS: The sixteen liver transplant models in rhesus monkey which were constructed successfuly by the method of “double-cuff and one support tube” were divided into two groups randomly: experiment group (no treated by immunosuppressant in perioperative period) and control group (treated by immunosuppressant in perioperative period). Then the blood specimen and liver tissue respectively were colected at 6, 12, 24 and 72 hours after operation. The levels of alanine transferase, aspartate aminotransferase, and total bilirubin were detected with the fuly automatic biochemical analyser. The levels of CD4+/CD8+were tested by flow cytometry. The liver tissue in rhesus monkey after liver transplantation was detected by hematoxylin-eosin staining. The degree of acute rejection was evaluated by Banff Score System. RESULTS AND CONCLUSION: Acute rejection appeared in the experiment group at 12, 24, and 72 hours after liver transplantation. Levels of alanine transferase, aspartate aminotransferase, and total bilirubin were significantly higher in the experimental group than in the control group at 24 and 72 hours after transplantation (P < 0.05). The expression of CD4+/CD8+of the experiment group and control group began to rise at 6 hours after surgery, but the experiment group increased the most obvious. CD4+/CD8+ expression was significantly greater in the experimental group than in the control group at 24 and 72 hours after transplantation (P < 0.05). Morphological pathology was severer, and Banff score was higher in the experiment group than in the control group at 72 hours (P < 0.05). These data suggested that the variation of expression of CD4+/CD8+was earlier than the change of liver tissue pathology and the change of liver function in the early acute rejection after liver transplantation. The rise of level of CD4+/CD8+ after liver transplantation indicated the increase of celular immunity in body, which had an important role in the early diagnosis of acute rejection after liver transplantation.

BACKGROUND:At present, the proteome is a mature technology that has been applied in basic research fields related to liver transplantation. But, it has been not reported in research related to reduced-size liver transplantation.
OBJECTIVE:To explore the expression of differential proteins related to hepatic energy metabolism fol owing reduce-size liver transplantation in rats by using by proteomic technology.
METHODS:The improved model of reduced-size liver transplantation was used in this experiment. The donor was health female Lewis rats and the recipient was male Wistar rats for liver transplantation. The difference between the donor and the recipient was about 20 g. The weight of donor liver/the weight of recipient donor was approximately equal to 50%. The donor liver tissue was harvested and trimmed to the required size. The portal vein and infrahepatic vena cava were cannulated, and the biliary tract was implanted into the donor bile duct for transplantation. Then the donor was transplanted into the recipient after the removal of original liver tissue. Hepatic specimens were harvested by 1, 3 and 7 days after reduced-size liver transplantation. Then, the harvested specimens were compared with the normal donor and recipient liver tissue that were previously harvested and frozen, to generate two-dimensional gel electrophoresis profile using proteome technology. Then tandem mass spectrometry and databases analysis were performed after two-dimensional electrophoresis for identifying differential protein stains.
RESULTS AND CONCLUSION:In this experiment, 72 differential protein stains with over lo-fold changes were selected. After identification, 32 proteins showed clear functions, and among them three differential proteins (ATP synthase beta subunit, electron-transferring flavoprotein beta peptide and proton-transferring ATP synthase) were involved in the process of cel energy metabolism. The proteins were distributed on 1 and 7 days after reduce-size liver transplantation, accounting for 6%.

Objective To explore the influential factor and management of biliary tract complica-tions after liver transplantation. Method Clinical data of 57 patients who underwent liver transplantation between May 2006 and November 2007 were studied retrospectively. Results Among the 57 patients, 8patients (14.04%) developed postoperative bililary tract complications,4 patients with biliary leakage, 2patients with anastomosis stricture, 1 patient with intrahepatic biloma, all above eases were fully recover, 1patient with anastomosis stricture and intrahepatie biliary tract casting mould received liver retransplantation.Conclusions The management of biliary tract complications after liver transplantation are difficult. To at-tach importance to influential factor and prevention, timely diagnosis and treatment will improve patients' sur-vival time and quality of life.