Laboratory Study Of Earlier Diagnoses Of The Pancreas Acute Rejection After Of The Simulataneous Pancreas And Kidney Transplantation In Pig

Section One The pig model establishment of portal refluenceand intestinal conduction in SPKObjective To establish the pig model of portal drainage and intestinal conduction in SPK.Methods Select 36 pigs of local hybridized pigs first generation, all age between 2 and 3 months. Then divide them into 2 groups equally by compatibility of the blood type, one group is donor and the other is recipient. 1 Operation for donor: the key is to establish hypothermic perfusion in good time in original position and shorten the ischemia time.This requires the familiarity with pig anatomy and take the measure of the together excision of multiorgan in abdomen after brain death. The warm ischemia time under the method that take the kidney and pancreas out together is 0,which is designed by ourselves; 2 Operation for recipient: Put a tube in the external jugular vein of the donors routinely for the convenience of observation during theoperation and transfusion and taking blood for test after operation. Excise the whole pancreas and the right kidney, inosculate pancreatic and portal veins end-to-side, inosculate nephric veins end-to-end, inosculate upper abdominal aorta with celiac artery, superior mesenteric artery and right nephric vein and abdominal aorta beneath the kidney. Inosculate duodenum and upper segment of jejunum end-to side. The pancreatic juice and urine are conducted out of the body; 3 Transfusion after operation, monitor the vital signs dynamically, and doing biopsy at fixed time.Results 18 cases underwent operation, one recipient died after operation for deep anaesthesia, the success rate is 94.4%. 1 died after the first biopsy, 2 died after the second biopsy, and 1 died at the sixth day after operation. The level of blood sugar,insulin and glucagon decreases after the excision of pancreas for 30 min (P<0.05) , pancreatic juice and urine outflow after operation, and the blood sugar is mormal and stable. Some recipients appear rejection in 3 days after operation, on the seventh day, all of them appear rejection . But the function of the implant have no obvious change (P>0.05). Conclusions When studying SPK, we adopt the method which excise the pancreas to establish IDDM .This is an ideal model of big animal, for its simplicity, maneuverability, stability, livability and repeatability. So, it is very useful for us to improve the operation and study its basic theory. Section 2 To study acute rejection of the pertinence amongkidney ,duodenum and pancreas in SPKObjective To study the value of the pathologic change in duodenum and kidney after SPK in the diagnosis of pancreas acute rejection. Methods Select 36 local hybridized pigs of the first generation. Divide them into 2 groups equally by compatibility of the blood type, one group is donor and the other is recipient. Test the endocritic function of the pancreas 1 day before operation and the third, fifth, seventh day after the operation. Do biopsy of pancreas, duodenum and kidney at the third, fifth, and seventh day after operation.Results 1 There is no significant change of the endocritic function of pancreas in earlier of the pancreas acute rejection(p>0.05); 2 Within 3 days, the number of acute rejection occur in pancreas duodenum and kidney are 5, 6 and 7 respectively, and that goes up to 12,13 and 13 on the seventh day after the operation. The correlation coefficient on the third, fifth day after the operation between duodeum and pancreas is 71.8%,83.2%,and the kidney is 64.8%,70.3% respectively; 3 The degree of the rejection vary, but there is no significant difference (P>0.05) .Conclusions In SPK, there is a high pertinence of pathologic change among biopsy of kidney, duodenum and pancreas in acute rejection, especially the duodenum and the pancreas. The acute rejection cannot be diagnosed via the biopsy of kidney or duodenum respectively, but it can be when together. The degree of the rejection vary, and the change of endocrinology lags behind the pathologic change obviously.Section3 The study of the effect of cytokine in earlierdiagnoses of pancreas acute rejection after SPK Objective To study the effect of the expression of IL-2 mRNA and protein in different tissue and the same tissue at different time in earlier diagnoses of the acute rejection after SPK.Methods Select 44 local hybridized pigs of the first generation. Divide them into 2 groups, the control group consists of 8, while the experimental group are devided into 2 groups equally by compatibility of the blood type, one group is donor and the other is recipient. The experimental group is operated with SPK, the pancreas and right kidney are dissociated in the control group, and the pancreatic juice and urine are conducted out of the body. The exposure time of abdominal cavity has no difference. Take the blood respectively 1 day before and 1, 3, 5, 7 days after the operation, test the endocritic function and the expression of mRNA and protein of IL-2 and IL-4 with RT-PCR and ELISA. Do biopsy of pancreas, duodenum and kidney , and test the expression of mRNA and protein of IL-2 and IL-4 in local aera. Results 1 There is no significant difference between the experimental and control group in change of the level of blood sugar, insulin and glucagon(P>0.05); 2 There is no significant difference between the experimental and control group of the change of IL-2 in serum the moment the operation was just finished, 1 and 3days after the operation(p>0.05), butthere is significant difference on the 5th and 7th day(p<0.05); 3 There is no significant difference of the level of IL-4 in all stage in pancreas(p>0.05); 4 There is significant difference between preoperation and 3 days after operation of the expression of mRNA and protein of IL-2 in local aera pancreas (p<0.05).Conclusions The change of IL-2 in local aera is a sensitive indicator for the earlier diagnose of acute rejection.. The change of IL-4 is not obvious in all stage, but it is expressessed dissimilarly in different tissue.