1.Effect of CaMK Ⅱ expression on apoptosis of rat hepatocytes BRL-3A
Jianghua RAN ; Kepu ZHENG ; Wang LI ; Xibing ZHANG ; Boqiang LIU
Chinese Journal of Organ Transplantation 2017;38(4):239-243
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.
2.Joint Effects of Selective Digestive Decontamination and Glutamine on Intestinal Bacterial Translocation of Piggyback Liver Transplantation in Rabbit
Li LI ; Zhu LI ; Jianghua RAN ; Gang CHEN
Chinese Journal of Bases and Clinics in General Surgery 2003;0(02):-
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
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.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.
5.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.
6.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).
7.Study on the combined application of laparoscopic cholecystectomy and endoscopic sphincterotomy in treatment of cholecystolithiasis and secondary choledocholithiasis
Daguang TIAN ; Jie HUANG ; Jie ZHANG ; Jianghua RAN ; Xiaoping WEI ; Guohai DONG
Chinese Journal of General Surgery 2001;0(08):-
Objective To study the therapeutic effects of combined use of laparoscopic cholecystetomy and endoscopic sphincterotomy for cholecystolithiasis with secondary choledocholithiasis.Methods Thirty-five patients were diagnosed as cholecystolithiasis with secondary choledocholithiasis by B-ultrasonography and magnetic resonance cholangiopancreatography.Of them,in 28 cases,laparoscopic cholecystetomy was performed first,and ERCP and endoscopic sphincterotomy were done one week later;in 7 cases,endoscopic sphincterotomy were performed before laparoscopic cholecystectomy.Results The outcome of all the thirty-five cases was satisfactory without severe complications or conversion into open procedure.Conclusions The method of combined laparoscopic cholecystomy and endoscopic sphincterotomy,for cholecystolithiasis with secondy choledocholithiasis,especially for cases in whom the diameter of the common bile duct stone is ≤1cm,can give good therapeutic results and has advantages of minimal invasiveness,few complications and quick recovery.
8.Improvements on Surgical Model Techniques of Rat Orthotopic Liver Transplantation
Qian WANG ; Li LI ; Jianghua RAN ; Xiaoyan LI ; Gang CHEN ; Hongxian YAN
Journal of Kunming Medical University 1990;0(02):-
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.
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.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.