Experimental Study On Auxiliary Partial Orthotopic Liver Transplantation For Wilson's Disease

PartⅠExperimentⅠEstablishment of orthotopic liver transplantation in miceObjective To improve the technique of orthotopic liver transplantation model in mice and build animal model on the study of transplantation immunity, liver regeneration and ischemical reperfusion injury.Methods According to "two-cuff" method,the surgical technic of harvesting donor's liver,fixing the cuff and the anastomosis of superior and inferior caval vein were improved.Results The operation time of harvesting donor's liver and anhepatic phase and recipient were (18±2) min, (17±2) min and (43±3) min.The model success rate was 92%,and the 100-day survival was 85%.Conclusions High quality liver could be harvested by this improved method,and short anhepatic phase ensured the high model success rate. Part I Experiment II Establishment of auxiliary partialorthotopic liver transplantation in miceObjective To explore the technique of auxiliary partial orthotopic livertransplantation model in mice and provied experimental animal data for clinicalapplication.Methods The donor portal vein and hepatic vein are anastomosed with Cuff andsutrue technique respectively. The donor bile duct is implanted into recipientduodenum.Results The operation time of harvesting donors liver and anhepatic phase andrecipient were (30士3) min, (6士1) min and (58士5) min. The model successrate was 96 %,and the 4-week survival was 88 %.Conclusions The animal model was stable with high success-rate and can be used forthe study of auxiliary partial orthotopic liver transplantation.Part II Experiment I Reproduction and genotypeidentilfcation of Atp7b-/- mouse and its disorder of copper metabolismObjective To explore the reproduction and genotype identification of Atp7b-/- mouseand its disorder of copper metabolism.Methods The heterozygous mice were used for breeding, and the genotypeidentification of offspring was used by PCR. Copper contents were determined byatomic absorption spectrometry and HE staining was made for examination ofhistopathological change.Results The analysis of genotype indentification fit the Mendel's laws. HepaticAcopper of Atp7b-/- mice were about 25 times more than wildtype, but serum copperincreased and peaked at 20 week old and were twice more than wildtype. At 20 weekold, hepatocellular necrosis, fibrosis and nodules were detected in almost all theAtp7b-/- mice. Conclusions The Atp7b-/- mice could be inherited stably, and their had significant hepatic histopathological change that can be an ideal experimental animal of Wilson's disease.PartⅡExperimentⅡExperimental study on auxiliary partial orthotopic liver transplantation for Wilson's diseaseObjective To evaluate the treatment effectiveness of auxiliary partial orthotopic liver transplantation for Wilson's disease.Methods Auxiliary partial orthotopic liver transplantations were performed at 8 and 20 weeks old and the Left lateral lobe was the donor liver which accounts for 38% of the total recipient liver. Atp7b-/- mice were recipients, and Atp7b+/+ mice were donors. The survival and index of copper metabolism and liver function were observed after operation.Results The postoperative survival of 8 and 20 weeks time point were 92% and 85% respectively. Serum copper, remanent liver copper, serum ceruloplasmin activity and liver function were improved postoperatively, and in 8 weeks time point kidney copper and brain copper content decreased after operation, meanwhile the pathological changes of remanent liver was slighter than control; in 20 weeks time point, the donor liver proliferated and the remanent liver atrophied after operation, and the smaller donor liver (19% of total recipient liver) could also corrected the disorder of copper metabolism as same as OLT, but had lower surgical risk.Conclusions Left lateral lobe auxiliary partial orthotopic liver transplantation could take the place of orthotopic total liver transplantation for Wilson's disease, and early transplantation performed, the pathological process of remanent liver delayed and copper deposition in other ograns decreased. However, the minimum volume of liver meet the body's copper metabolic needs still needed to be further studied.