Objective To observe the dynamic histopathologic changes of acute rejection in rat orthotopic liver transplantation (OLT) model after tacrolimus discontinued and provide some prediction and evaluation data for clinical acute rejection after liver transplantation. Methods Kamada two-cuff technique was used to establish 60 rat OLT model， and male DA rats, male Lewis rats were used as donors and recipients respectively. Therapeutic amount of tacrolimus (0.05 mg/kg, twice per day, continued for 8 d, 1 d before operation and 7 d after operation, intragastric administrated) was administrated to recipients, then continuously half dose was decreased every day beginning from day 8 after operation and tacrolimus administration was stopped on day 13. Liver tissues were collected on day 7, 14, 21, and 28 after liver transplantation. Histopathologic changes and rejection activity index (RAI) of liver tissues were observed, survival time of recipients was calculated. Results Owing to protection effects of tacrolimus, liver tissues displayed no significant histopathologic changes of acute rejection in 7 d after OLT, while typical acute rejection histopathologic changes began to be observed on day 14 after OLT due to tacrolimus discontinuation. On day 14, 21, and 28, RAI were 3.7±0.9, 6.3±0.9, and 8.1±0.7 respectively. Survival time of recipients was (20.85±0.71) d with a median of 21 d. Conclusion Acute rejection could be induced in rat OLT model after tacrolimus discontinuation, and data collected from this model shows some extent of predictive value and assessment value for clinical liver acute rejection.
【Abstract】ObjectiveTo investigate the risk factors for acute lung injury(ALI) after orthotopic liver transplantation(OLT) and to explore the prevention and cure scheme.MethodsThe risk factors responsible for ALI in 4 patients undergoing OLT were analyzed with retrospective investigation.ResultsPortal pulmonary hypertension, longterm mechanical ventilation, severe infection, SIRS, hypercoagulability, overdose transfusion and kidney dysfunction were risk factors for ALI.ConclusionALI frequently occurred after OLT. Reducing and diminishing the risk factors is very important to avoid ALI after OLT.
Objective To summarize the changes of serum ceruloplasmin levels and urinary copper excretion in Wilson’s disease （WD） after living-related liver transplantation(LRLT) and orthotopic liver transplantation(OLT). Methods From September 2000 to November 2003, 140 cases liver transplantation were performed in our Liver Transplantation Center, LRLT and OLT were carried out in 26 patients with WD, three of them had fulminant hepatic failure and the others had end-stage hepatic insufficiency. All the recipients had low serum ceruloplasmin levels 〔(124.8±22.8) mg/L〕 and high urinary copper excretion 〔(1 524.8±328.6) μg/24h〕 before transplantation. The serum ceruloplasmin levels and urinary copper excretion were within normal limits in 22 donors 〔(230.4±29.6) mg/L〕 and ＜50 μg/24h〕. Results All recipients recovered satisfactorily. After operation 1, 3, 6,12 months, in OLT group, serum ceruloplasmin level and urinary copper excretion were (320.2±36.8) mg/L, (380.4±45.6) mg/L, (360.5±37.6) mg/L, (356.2±27.6) mg/L and (240.4±22.8) μg/24h, (86.5±10.6) μg/24h, (54.2±6.8) μg/24h, (46.8±3.4) μg/24h; While in LRLT group, serum ceruloplasmin levels and urinary copper excretion were (216.8±20.4) mg/L, (248.5±32.6) mg/L, (285.4±44.3) mg/L, (260.2±36.6) mg/L and (380.8±37.6) μg/24h, (150.6±24.5) μg/24h, (75.5±9.6) μg/24h, (60.3±5.8) μg/24h. Conclusion OLT and LRLT are curative procedure in WD manifested as fulminant hepatic failure and/or end-stage hepatic insufficiency. After liver transplantation, the serum ceruloplasmin level can increase to its normal range while urinary copper excretion decreases.
ObjectiveTo explore the application of extracorporeal venovenous bypass in orthotopic liver transplantation in pigs and to compare hemodynamic changes during operation of two different bypass ways. MethodsTwentyfive porcine orthotopic liver transplantations were performed and extracorporeal venovenous bypass was established during anhepatic phase through a catheter in portal vein (group A,n=16) or in splenic vein (group B,n=9).Hemodynamic changes were monitored continuously.ResultsFourteen recipients survived two days after operation (14/16) in group A while all survived in group B (9/9).Transient hemodynamic disturbance (MAP and CVP decreased,and HR increased) was monitored at both the beginning and the end of anhepatic stage in group A,while these parameters kept stable in group B (P＜0.05).ConclusionApplying venovenous bypass may stabilize recipients’ hemodynamics in porcines orthotopic liver transplantation,and splenic vein draining way has more advantages than portal vein.
ObjectiveTo evaluate the changes in thrombelastography(TEG) during orthotopic liver transplantation （OLT） in Chinese. MethodsTwentyfive patients with cirrhosis of liver undergoing OLT were studied. They were composed of two groups: cirrhosis group （n=15） and liver neoplasm group （n=10）. Anesthesia was induced with propofol 1.5-2 mg/kg,fentanyl 3-5 μg/kg and vecuronium 0.1 mg/kg and maintained with isoflurane or enflurane inhalation.The operation was divided into three phases: ① before operation and preanhepatic phase （120 min after operation was started）, ② 30 min after liver was removed,③ 5 min before reperfusion and 5 min,15 min,30 min,60 min and 120 min after reperfusion.In 8 patients among the 25 patients heparinasecelite TEG was measured 5 min after reperfusion in addition to celite TEG.If there was significant differences in traces between the two TEG measurements,an intravenous bolus of 50-75 mg protamine was given and the heparinasecelite TEG was repeated.The measured variables included the r （reaction） time,representing the rate of initial fibrin formation K （coagulation） time, alpha angles （α） reflecting fibrinplatelet interaction, MA （maximal amplitude） indicating qualitative platelet function and percent fibrinolysis at 60 min. ResultsIn cirrhosis group changes in TEG occurred after liver was removed and in earlier period after reperfusion, while in liver neoplasm group changes in TEG were found in earlier period after reperfusion as compared with preoperative value.At 5 min after reperfusion there were significant differences in TEG （r，K，α and MA） values between celite and heparincelite TEG （P＜0.01）. ConclusionDuring OLT coagulation disorder occurs mainly at anhepatic and early reperfusion phase.
ObjectiveTo evaluate the application of portosystemic shunt after subcutaneous transposition of the spleen (STS) to orthotopic liver transplantation (OLT) in the rat. MethodsOne hundred and eighty Wistar rats were randomly divided into the group of orthotopic liver transplantation after portosystemic shunts by subcutaneous transposition of the spleen (STS+OLT group) and the group of orthotopic liver transplantation (OLT group). The two groups were further divided into A, B, C subgroups in the light of duration of anhepatic phase (phases were respectively less than 25 min, around 35 min and 45 min).There were fifteen rats in each subgroup. At the described intervals, blood samples were collected from the peripheral and portal vein for testing ALT, pH and endotoxin levels. The survival rates were also observed. Results The ALT value of all animals basically returned to normal levels on the 7th postoperative day in the STS+OLT group and the OLT A subgroup, but in OLT B subgroup, ALT was still remarkably elevated on the 7th postoperative day (P＜0.01), and returned to normal levels on the 30th postoperative day. The pH values and endotoxin levels from the portal vein of all animals in STS+OLT groups and OLT A subgroup had no significant difference (Pgt;0.05) at the beginning, the end of the anhepatic phase and at the time of reperfusion for 30 min. But in the OLT B and C groups, the pH values and endotoxin levels were significantly higher at the end of anhepatic phase and reperfusion for 30 min than those in the beginning of anhepatic phase (P＜0.01). The survival rates at postoperative different time points in both B and C subgroup of the OLT group were significantly lower than those in STS+OLT group animals (P＜0.05).Conclusion The portosystemic shunt by subcutaneous transposition of the spleen can notably improve both the success rate of the OLT procedure and the postoperative survival rate in the rat.
【Abstract】Objective To investigate the result of liver transplantation for end stage liver disease. Methods A retrospective analysis was made for 7 cases orthotopic liver transplantation(OLT) and 4 cases living related liver transplantation (5 patients with hepatitis B cirrhosis and 6 with Wilson’s disease),cirrhosis group was treated with lamivudine plus low dose anti-HBV-Ig. Results Ten patients were completely recovered discharged(including 4 cases LRLT) and only 1 died of ARDS.The complications after operation were: 2 cases of abdominal hemorrhage,3 cases of acute respiratory distress syndrome; and 4 cases of hepatitis B cirrhosis were HBV-DNA(-) after operation.Copperoxidase in all with Wilson’s disease became normal. Conclusion Liver transplantation is effective measure for end stage liver disease and living related liver transplantation is suitable for the present medical condition in China.Surgical technique is crucial for reducing perioperative complications.
Liver transplantation in hamster-to-rat is a good model for the research in xenograft, but it is difficult to be performed. In order to simplify its procedures, 38 cases of liver transplantation in hamster-to-rat were performed with several technique improvments on the basis of orthotopic liver transplantation in rats. In the light of hamster’s anatomy, three cuffs anastomosis method was used. Because of its high stability and high survival rate, the model can be used widely as the research for liver xenograft.
ObjectiveTo explore the surgical techniques of rat orthotopic liver transplantation (OLT) model in rats using modified technique by single operator. MethodsA total of two hundreds cases of rats (one hundred pairs) were used to establish OLT model including eighty cases of rats (forty pairs) used in formal study by using modified Kamada “two-cuff” method, of which the key surgical procedures were further modified. Intraoperative results and postoperative recovery were analyzed. ResultsAll eighty cases of rats successfully operated. Operative time in donors was (28.5±2.4) min and reparative time of donor liver was (10.2±1.8) min. Anastomosis time for suprahepatic inferior vena cava and portal vein (15.3±1.9) min and (3.4±1.2) min, respectively. Anhepatic phase of transplant recipients was (23.8±1.9) min, anastomosis time for infrahepatic inferior vena cava was (5.1±2.1) min, and biliary reconstruction time was (3.1±0.9) min. Blood loss of donor and recipients during the operation was blew 0.5 ml. No recipient died for operation. The 2 d survival rate of recipients was 90% (36/40), three rats died of bleeding at the seam in hepatic inferior vena cava and one rat died of longer anhepatic phase. The 7 d survival rate of recipients was 82.5% (33/40), three rats died of intra-abdominal infections. ConclusionThe modification in en bloc separation of abdominal aorta and interior vena cava for abdominal aortic perfusion of donor liver, careful dissection of left subphrenic vein triangle and the hemostasis by heat coagulating can reduce operative complications, guarantee the quality of donor liver and improve the survival rate of rats in establishment of OLT model in rats using modified technique by single operator.
Objective To investigate the expression of transcription factor Foxp3 in the orthotopic liver transplantation by using the inbred rats with spontaneous immune tolerance. Methods The model of orthotopic liver transplantation was established on inbred rats according to double-sleeve technique. The total RNA that was isolated from liver was reversely transcribed into cDNA. The method of real-time fluorescence quantitative PCR (RFQ-PCR) was used to analyze the expression level of Foxp3 mRNA in tolerance group and syngeneic group, respectively. The expression of Scurfin in hepatic tissue was assayed by Western blot and then was analyzed by computer imaging system. Results The expression levels of Foxp3 mRNA and Scurfin in the transplanted liver were significantly lower than those of normal liver within the first week after transplantation. The level of Foxp3 mRNA began to increase on day 7 and reached the peak point on day 14. The expression level of Foxp3 mRNA began to decrease on day 30 but was still higher than the normal value (P＜0.05). The Western blot showed resemble changes on that of Scurfin. Conclusion Transcription factor Foxp3 may play an important role in the spontaneous immune tolerance in the orthotopic liver transplantation of inbred rat.