1.Expression of hepatic signal transduction proteins following reduced-size liver transplantation in rats
Jing LIU ; Li LI ; Jianghua RAN ; Shengning ZHANG ; Laibang LI ; Yang GAO ; Yiming CHEN ; Xibing ZHANG
Chinese Journal of Tissue Engineering Research 2014;(49):7974-7978
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.
2.Variation of T cell subset during acute rejection after liver transplantation in rhesus monkeys
Jianghua RAN ; Jing LIU ; Xibing ZHANG ; Shengning ZHANG ; Shuyuan WU ; Laibang LI ; Wang LI ; Li LI
Chinese Journal of Tissue Engineering Research 2014;(49):7948-7954
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.
3.Cause analysis of early death after liver transplantation in rhesus monkey
Jianghua RAN ; Jing LIU ; Shengning ZHANG ; Zhu LI ; Shuyuan WU ; Yu LIANG ; Xibing ZHANG ; Li LI
Chinese Journal of Tissue Engineering Research 2011;15(18):3413-3416
BACKGROUND: Various factors contribute to the establishment of liver transplantation models in rhesus monkey, the rate of successful operation and long-term survival are very low. OBJECTIVE: To analyze the cause of early death following liver transplantation in rhesus monkey. METHODS: Liver transplantation models were fabricated with the classical and modified methods in rhesus monkeys. Operation of donor was performed quickly by a big crucial incision of abdomen. The improved double-cuff of the portal vein and inferior vena cava were finished, in addition to stay pipe of biliary tract in the process of repairing donor liver. Operation of the receptor was performed by classical orthotopic liver transplantation. RESULTS AND CONCLUSION: A total of 25 pairs of rhesus monkeys were successfully for establishing liver transplantation models. Seven rhesus monkeys died within early stage of post-operation, including six out of nine monkeys died by using the classical approach and one out of sixteen monkeys died by using the improved approach. There were five of seven monkeys died of intra-abdominal hemorrhage, one died of primary graft nonfunction and one died of respiratory failure. Results indicated that, the major death cause after classical orthotopic liver transplantation in rhesus monkey is abdominal hemorrhage. The improved methods of liver transplantation apparently reduce the hemorrhage and raise early survival rate following liver transplantation.
4.Expression of hepatic energy proteins following reduced-size liver transplantation in rats
Jing LIU ; Li LI ; Jianghua RAN ; Shengning ZHANG ; Laibang LI ; Xibing ZHANG ; Yang GAO ; Yiming CHEN
Chinese Journal of Tissue Engineering Research 2015;(18):2874-2878
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%.
5.Hepatitis B virus recurrence after liver transplantation: Prevention and cure efficacy of nucleoside anti-hepatitis B medicine combined with low-dose anti-hepatitis B immunoglobulin
Jing LIU ; Shengning ZHANG ; Zhu LI ; Laibang LI ; Jianghua RAN ; Li LI
Chinese Journal of Tissue Engineering Research 2010;14(5):811-814
BACKGROUND: It was reported from home and abroad that the effect of nucleoside anti-hepatitis B medicine and anti-hepatitis B immunoglobulin for prevention and cure of hepatitis B virus recurrence after liver transplantation with hepatopathy correlation with hepatitis B was good for patients. But the reported dosage of anti-hepatitis B immunoglobulin in and after liver transplantation was different. OBJECTIVE: To verify and investigate the effect of nucleoside anti-hepatitis B medicine combined with anti-hepatitis B immunoglobulin on prevention and cure of hepatitis B virus recurrence after liver transplantation, METHODS: A retrospective analysis was performed on 59 patients with liver transplantation of hepatopathy correlated with hepatitis B who were selected from Liver Transplantation Center, the Ganmay Affiliated Hospital of Kunming Medical College between May 2006 and February 2009. A total of 50 out of 59 cases were diagnosed with posthepatitic cirrhosis in decompensatio stage before transplantation, including 15 cases having positive hepatitis B DNA. Before liver transplantation, 5 cases accepted Lamivudine, 1 case accepted Adefovir dipivoxil, and 1 case accepted Entecavir. Treatment time ranged from two weeks to one year. All the patients accepted intramascular injection of anti-hepatitis B irnrnunoglobulin, 200 U/d; which were adjusted in the light of hepatitis B surface antibody titer. A total of 55 out of 59 cases accepted Lamivudine, 3 cases accepted Adefovir dipivoxil, and 1 case accepted Eetecavir after liver transplantation. RESULTS AND CONCLUSION: Two patients underwent hepatitis b virus reinfection, but HBV variants (YMDD) reinfection was not determined, one of which occurred in one year after liver transplantation with positive pre-OLT serum hepatitis b virus DNA, another after one year with negative pre-OLT serum hepatitis b virus DNA. The reinfection rate of group with negative or positive pre-OLT serum HBV DNA was 2% and 7%, respectively, it was maybe well prevention and cure of hepatitis B after liver transplantation that patients accepted nucleoside anti-hepatitis B medicine combined with low close anti-hepatitis B immunoglobulin (200 U/d).
6.Influential factor and management of biliary tract complications after liver transplantation
Shengning ZHANG ; Li LI ; Jianghua RAN ; Xiaoyan LI ; Zhu LI ; Jing LIU ; Xiaoping WEI ; Laibang LI
Chinese Journal of Postgraduates of Medicine 2008;31(11):17-20
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.
7.Modified model of reduced-size liver transplantation in rats
Jing LIU ; Jiang LI ; Shengning ZHANG ; Zhu LI ; Laibang LI ; Jianghua RAN ; Li LI
Chinese Journal of Tissue Engineering Research 2010;14(18):3252-3257
BACKGROUND: There are few studies on liver regeneration following living liver transplantation. Improvement of operation methods and techniques and successful rate are the basis for rat liver transplantation study and data acquisition. OBJECTIVE: To investigate the efficacy of improved model of reduced-size liver transplantation in the rat. METHODS: Healthy SD rats were selected. 70 pairs of rats were subjected to reduced-size liver transplantation before modification, and 100 pairs subjected to reduced-size liver transplantation after modification. The donors were female and the recipients were male, and the body mass of donors was 10 g less than the recipients. Operation of donor was performed by only one person with the naked eye, and reduced-size donor liver was performed in the donor operation. The handle of self-made cannula was placed in the front of portal vein and inferior vena cava, respectively, and the tied ligature of pyloric veins was turned inside out of the self-made cannula. Furthermore, the tied ligature was placed in the left of the self-made cannula; the same to inferior vena cava except that the tied ligature of right renal vein was placed in the right of the self-made cannula; the portal vein and inferior vena cava were washed with self-made perfusate respectively. Operation of the receptor was performed by two persons with the naked eye, with improved dual-cuff technique of Kamada and stay pipe of biliary tract, the fixed points of left and right were connected by anastomosis of "8" type with turning inside out while inosculating inferior vena cava. RESULTS AND CONCLUSION: The average modified operation time of the donor and the donor liver preparation time was (32±2) minutes and (6±2) minutes, respectively. The average operation time of the recipient and the anhepatic time was (40±3) minutes and (14±3) minutes, respectively. The general successful rate was 92%; three-day survival rate was 85% and two-week survival rate was 83%. The postoperative complications reduced significantly (P< 0.05), and cold conservation time of donor was shortened (P < 0.05). The modified model of reduced-size liver transplantation was more safe and reliable, with high success rate of liver transplantation and survival rate of recipient. Moreover, the postoperative complications of receptor decreased significantly. It provide an effective method of investigating liver graft regeneration following reduced-size liver transplantation.
8.Mechanism of Immune Hyporesponsiveness Induced by Recipient-Derived Immature Dendritic Cells in Rat Liver Transplantation
Li LI ; Shengning ZHANG ; Jianghua RAN ; Jing LIU ; Zhu LI ; Yongheng ZHAO ; Laibang LI
Chinese Journal of Bases and Clinics in General Surgery 2004;0(01):-
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
9.Effect of rhGH on the Protein Expression of NF-?B P65 in Ischemic Reperfusion Injury of Rat Liver
Jianghua RAN ; Yongzhang GUO ; Li LI ; Shengning ZHANG ; Zhu LI ; Jing LIU
Journal of Kunming Medical University 1986;0(04):-
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(0.2IU/100g 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
10.Improvements of Orthotopic Liver Transplantation Model in Rhesus Monkey
Jianghua RAN ; Shengning ZHANG ; Jing LIU ; Zhu LI ; Laibang LI ; Shuyuan WU ; Li LI
Chinese Journal of Bases and Clinics in General Surgery 2008;0(07):-
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.