Abstract:Objective To investigate the expression and significance of Voltage-gated Cl channel-3 （ClC-3） in acute cardiac allograft rejection in rats. Methods The model of heterotopic cardiac allograft of SD to Wistar rats was established. The rats were divided into two groups： control group and cyclosporin A（CsA） treated group （CsA group）. Living span of the transplants in eight rats of each group were observed. Allograft samples were harvested separately on the day 1, 3, 5, 7 after operation （n = 6）. The rejection was evaluated by routine pathological examinations. The myocardial apoptosis by terminal deoxylnucleotidyl transferase mediated-dUTP nick end labeling （TUNEL） method and the local expression of ClC-3 were detected by reverse transcriptase polymerase chain reaction （RT-PCR）. Results The allografts survival time was significantly longer in CsA group compared with that in control group （15.4±5.1dvs. 7.6±1.5d, P〈0.05）. There was lesser pathological changes in CsA group than that in control group. The apoptosis index were significantly higher in control group and the expression of ClC-3 was significantly lower（P〈0.05）. CsA could inhibit the rise of apoptosis index and the decrease of the ClC-3 expression. Conclusion The ClC-3 expression is closely related with the severity of myocardial necrosis and apoptosis index, which indicates that ClC-3 plays a very important role in the necrosis and apoptosis during acute cardiac allograft rejection of rat.
Objective To insure early detection and hence efficient prevention of allograft rejection in transplanted heart, investigate possible applications of NAD(P)H fluorescence components analysis at the level of living cardiac cells to propose new approaches for diagnosis of rejection. Methods NAD(P)H was studied for noninvasive fluorescent probing of the mitochondrial function. Human cardiomyocyte were isolated from one additional endomyocardial biopsy (EMB) of 14 pediatric patients with heart ransplantation. Rat cardiomyocyte (n=5, 13-14 week old) were also isolated by the same approach for human myocytes. Autofluorescence(AF) was recorded in living cardiomyocytes following excitation with 375 nm UVlight and detection by spectrallyresolved time correlated single photon counting (TCSPC), based on the simultaneous measurement of the fluorescence spectra and lifetimes. Rat cardiac cells were divided into four groups: normoxic condition, normoxia with Rotenone, ischemic condition and ischemia with Rotenone. Comparison of cardiomyocyte AF between human and rat; compared kinetics of rat cardiomyocytes AF in normoxic conditions to ischemiamimicking ones, induced at physiological temperatures by reducing cell pH and oxygen content; comparison of cardiomyocyte AF dynamic changes in transplanted pediatric patients presenting either no rejection (R0) or mild rejection (R1). Results We have achieved appropriate isolation of living cardiomyocytes from human biopsies, as well as from rat cardiac tissues and determined their AF. At least a 3-exponential decay with 0.5-0.7ns, 1.9-2.4 ns and 9.0-15.0 ns lifetime pools is necessary to describe human cardiomyocyte AF within 420560 nm spectral range. Rat cardiomyocyte steadystate AF in ischemiamimicking condition was significantly increased when compared normoxic ones (Plt;0.05); application of Rotenone induced a significant increase in AF intensity in ischemic and normoxic condition, however no significant difference between the two groups (Plt;0.05).Human cardiomyocyte AF was found significantly lower in comparison to experimental rat model in the same condition(Plt;0.05). A correlation between changes in steadystate NAD(P)H fluorescence and rejection grades was found when comparison of R1 to R0. R1 showed significantly increased fluorescence intensity (Plt;0.05), without change in the spectra shape, results can be comparable to the effect of ischemiamimic conditions. Conclusion Our studies clearly demonstrated that spectrallyresolved fluorescence spectral analysis coupled to fluorescence lifetime are high sensitive approaches to examine mitochondrial metabolic oxidative state directly in living human cardiomyocytes with good reproducibility. Human cardiomyocytes are more metabolically active than the rat ones, while this activity (and thus ATP production) seems lowered during rejection process. In perspective, the advantage of this method is the possibility of its combination to multiphoton confocal microscopy, which can result in the adaptation of this approach directly to tissue biopsy, as well as in vivo directly via cardiac catheterization without the necessity of cell isolation. This approach provides promising new tool for clinical diagnosis and treatment of allograft rejection, and will enhance our knowledge about cardiomyocyte oxidative metabolism and/or its dysfunction at a cellular level.
Objective To summarize the clinical experience of liver retransplantation. Methods Six liver retransplantations were performed. The indications consisted of primary non-function (PNF, 2 cases), acute or chronic rejection (2 cases), stomas stenosis of biliary tract (1 case) and primary sclerosing cholangitis (1 case). The immunosuppressive protocols included tacrolimus, methylprednisolone (MP) and mycophenolate mofetil (MMF). Results Five patients were cured. One patient died on day 4 after liver retransplantation because of multiple organ failure. Postoperative complications included deep fungal infection and wound infection. Conclusions Liver retransplantation is an effective method for graft failure after liver transplantation. Proper indication and optimum operative time, intensive perioperative supervision and proper treatment are very important to improv effect of liver retransplantation.
【Abstract】Objective To evaluate the pathological diagnosis of liver allograft rejection. Methods Literatures about diagnosis of liver transplantation rejection in recent ten years were reviewed.Results Humoral rejection was rare. The main features were graft blood vessel thrombosis and liver damage and necrosis about some days or one week after transplantation. The humoral rejection of liver graft occurred later than that of kidney and heart transplantation. The diagnosis of acute liver graft rejection was based on Banff Schema. During chronic rejection intrahepatic bile ducts among hepatic lobules in portal area disappeared, and inflammation, fibrosis and stricture of hepatic arteries and veins were found, but the morbidity was less than that of kidney， heart, lung and pancreas grafting. Conclusion Banff standard is the most important base of diagnosing liver graft rejection.
ObjectiveTo compare tacrolumus (FK506) with cyclosporine A (CsA) in clinical application to organ transplantation.MethodsThe literature in recent years has been reviewed and compared. ResultsFK506 was a powerful immunosuppression with a mechanism of action similar to that of CsA, but significantly superiori to CsA in terms of prophylaxis and treatment of allograft acute rejection, delay of chronic rejection, and withdrawal of steroid in early period. The cardiovascular mortality and chronic graft nephropathy (CGN),such as hypertension and hyperlipidemia were less frequently seen in FK506treated patients and FK506 also had an acceptable safety profile, including a low incidence of hypertrichosis,gingival hyperplasia and infections.However, CsA had been showed a better result in prevention of posttransplantation diabetes mellitus (PTDM ) and more economic agent than FK506. Pharmacokinetic studies showed CsA in the form of Sandimmun Neoral showed less inter an intrapatient variability than FK506.Meanwhile, the combination of MMF and FK506 or CsA has been proved effectively with excellent graft and patients survival. Conclusion FK506 and CsA are safe and effective long term maintenance immunosuppressive agents in organ transplantation with wonderful prospect.
Objective To investigate the effect of interleukin-10 （IL-10） gene transfer on expression of CD44, selectin-E, lymphocyte function associated antigen-1 （LFA-1）, vascular cell adhesion molecule-1 （VCAM-1） in mice heart transplantation rejection. Methods Model of mice cervical heterotopic heart transplantation was set up, 96 mice were divided into three groups with random number table, control group： heart transplantation between C57 mice; transplant group： heart from BALB/C mice transplant to C57 mice; IL-10 group： IL-10 was transfected on BALB/C mice isolated heart for 1 hour, then transplanted to C57 mice. The messenger ribonucleic acid （mRNA） level expression of CD44 ,selectin-E ,LFA-1 ,VCAM-1 and IL-10 were measured by reverse transcription-polymerase chain reaction （RT-PCR） at the 5th day after transplantation. Results The mRNA level expression of CD44, selectin-E ,LFA-1 ,VCAM-1 in transplant group were significantly increased than those in control group （P〈0.01）. The mRNA level expression of CD44, selectin-E, LFA-1 ,VCAM-1 in IL-10 group were significantly decreased than those in transplant group （P〈0.01）. Conclusion IL-10 gene transfer is able to decrease the expression of CD44, selectin-E,LFA-1 ,VCAM-1 and suppress the heart transplantation rejection in mice.
Objective To establish the model of pancreatoduodenal allotransplantation in pigs with enteric drainage (ED) and portal venous drainage (PVD). Methods Forty-six hybrid landraces were divided into two groups (donor and recipient groups) randomly, for pancreatoduodenal allotransplantation. Donors were perfused via abdomial aorta without clamping the portal venous outflow with UW solution after heparinization. Whole pancreatoduodenal graft was arvested with segments of abdomial aorta and portal vein and shaped under cold UW solution. Then, the end-to-end nastomosis was performed with the donor iliac artery bifurcation “Y” graft to the recipient superior mesenteric arteries and celiac artery. Furthermore, type Ⅰdiabete model was made by removal of the recipient pancreas. The venous anastomosis was reconstructed between the donor portal vein and the recipient superior mesenteric vein. Meanwhile, the end-to-side anastomosis was performed with the donor common iliac artery bifurcation “Y” graft to the recipient abdomial aorta and the side-to-side intestinal anastomosis was performed between the donor duodenum and the recipient jejunum. External jugular vein was intubated for transfusion. The levels of blood glucose， insulin and glucagon in blood were measured before and during the operation and 1， 3， 5， 7 d after operation. Results Twenty-three cases of pancreatoduodenal allotranplantations were performed on pigs. One died from complication of anesthesia. Success rate of operation was 95.7％.Complications of operation happened in 2 cases in which one was phlebothrombosis, incidence 4.5％and the other was duodenojejunal anastomotic leak, incidence 4.5％. The level of blood glucose increased within 30 min and recovered on the 2nd day after removal of pancreas. The levels of insulin and glucagon decreased within 30 min and recovered on the 2nd day after removal of pancreas. Rejection curred at the 1st day and reached the worst level on the 9th day after transplantation without the change of insulin and glucagon in blood and clinical symptoms of rejection. Conclusion Pancreatoduodenal transplantation in pigs can treat type Ⅰ diabete. ED and PVD can keep the function of endocrine in normal. The technique of duodenal transplantation with ED and PVD may pave the way for the further development of pancreas transplantation in clinic.
【Abstract】Objective This study was conducted to build experimental model of orthotopic liver transplantation in rat (ROLT) with the character of acute rejection; and to study the effect of cytotoxic T lymphocyte antigen 4 immunoglobulin G (CTLA4Ig) on prevention of rejection and the induction of immune tolerance of ROLT. Methods Build model of Wistar→SD ROLT(performed by the two cuff method) with character of acute rejection.Recipients were injected with CTLA4Ig 75 μg per ROLT into abdominal cavity after 2 days of operation. Contrast was made with no treatment group, the clinical characters, the liver function, the transplantated liver pathologic character and the concentrations of TNFα in serum were observed and measured on postoperative day 7. In treatment group, all above observation were done on postoperative month 4. Above all, determination of the effect of CTLA4Ig on preventing acute rejection and inducing tolerance in ROLT was observed.Results ①Recipients (no treatment group) died one by one within 6th～14th days; pathologic character of rejection in transplantation liver could be found; ② In treatment group, on postoperative day 7 and month 4, no clinical rejection character and no pathologic character of rejection in transplantation liver were found and serum concentration of cytokins related to TNFα found lower than that of contrast group(P＜0.05), and serum concentration of ALT、AST、TBIL、DBIL found lower too(P＜0.05); But serum concentration of TP and Alb was found higher than that of contrast group(P＜0.05). Conclusion ① CTLA4Ig treatment alone inhibits the rejection responce in ROLT. ② CTLA4Ig treatment can Prevent rejection and induce immune tolerance in ROLT model with characters of acute rejection; the serum concentration of cytokins related to TNFα can probably be used as evaluation standard of rejection in ROLT rejection.
Objective To investigate the effect of nidus vespae on lymphocyte blastisation in mixed culture system of lymphocyte and pancreatic islet. Methods Solution of nidus vespae was extracted with ethanol from its herb. Rat lymphocyte and pig pancreatic islet were isolated and then were mixed together and cultured in incubators of 37 ℃ (concentration in volume: 5%CO2). Three different concentrations of extracted solution of nidus vespae （experimental group Ⅰ: 4.0 g/ml, group Ⅱ: 0.4 g/ml, group Ⅲ: 0.2 g/ml） were added to the mixed culture system of lymphocyte, respectively. Radioactive nuclide counts per minute were measured by 3H-thymdine test in order to examine the role of nidus vespae in inhibiting blatisation of lymphocyte, and the results were also compared with control group and CsA group, respectively. Results The counts were all reduced significantly (P＜0.001) in 3 experimental groups compared with control group (group Ⅰ 45.3%, group Ⅱ 29.6%and group Ⅲ 9.2%). It also showed that the inhibitory effect became ber in higher concentration but still weaker than that in CsA group (80.7%). Conclusion Nidus vespae could inhibit the blastisation of lymphocyte in mixed culture system of lymphocyte and pancreatic islet, and the effect increased as the the concentration increased, which may suggest that nidus vespae could suppress the rejection induced by T cells.
ObjectiveTo reporte the nursing experience of non-healing incision due to allograft rejection after osteosarcoma surgery. MethodsOne patient with non-healing incision due to allograft rejection after osteosarcoma surgery treated in September 2013 was selected. The suitable moist healing dressings was chosen to control inflammation, prevent infection, manage exudation, promote the growth of granulation, protect the surrounding skin, shorten the dressing time and reduce the suffering of patients. ResultThe wound healed well after 65 days of dressing with the function of the right upper limb recovered. ConclusionThe moist healing dressing not only improved the quality of patient's life and increased the patient's confidence of overcoming the disease, but also made the patients more active to cooperate in the next treatment.