AIM:To investigate the impact of different time points of secondary warm ischemia on bile duct in a rat autologous liver transplantation model with external bile drainage.METHODS:One hundred and thirty-six male inbred...AIM:To investigate the impact of different time points of secondary warm ischemia on bile duct in a rat autologous liver transplantation model with external bile drainage.METHODS:One hundred and thirty-six male inbred SD rats were randomly assigned to one of four groups(Ⅰ-Ⅳ) according to the secondary warm ischemia time of 0,10,20 and 40 min.A rat model of autologous liver transplantation with continuous external biliary drainage under ether anesthesia was established.Ten rats in each group were used to evaluate the one-week survival rate.At 6 h,24 h,3 d and 7 d after reperfusion of the hepatic artery,6 rats were killed in each group to collect the blood sample via the infrahepatic vena cava and the median lobe of liver for assay.Warm ischemia time of liver,cold perfusion time,anhepaticphase,operative duration for biliary external drainage and survival rates in the four groups were analyzed for the establishment of models.RESULTS:No significant difference was shown in warm ischemia time,anhepatic phase and operative duration for biliary external drainage among the four groups.Five of the 40 rats in this study evaluated for the one-week survival rate died,including three deaths of severe pulmonary infection in group Ⅳ.A significant decrease of one-week survival rate in group Ⅳ was noted compared with the other three groups.With the prolongation of the biliary warm ischemia time,the indexes of the liver function assessment were significantly elevated,and biliary epithelial cell apoptosis index also increased.Pathological examinations showed significantly aggravated inflammation in the portal area and bile duct epithelial cell injury with the prolonged secondary warm ischemia time.Microthrombi were found in the micrangium around the biliary tract in some sections from groups Ⅲ and Ⅳ.CONCLUSION:The relationship between secondary warm ischemia time and the bile duct injury degree is time-dependent,and 20 min of secondary warm ischemia time is feasible for the study of bile duct injury.展开更多
Background Blocking the 4-1BB/4-1BB ligand （4-1BBL） signal may modulate the secretion of Th1/Th2 cytokines and prolong the survival of the grafts, which play a key role in organ transplantation tolerance. The aim of...Background Blocking the 4-1BB/4-1BB ligand （4-1BBL） signal may modulate the secretion of Th1/Th2 cytokines and prolong the survival of the grafts, which play a key role in organ transplantation tolerance. The aim of this study was to investigate the role of blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody （mAB） in acute rejection of rat orthotopic liver transplantation. Methods The orthotopic liver transplantation model was set up, while male Lewis rats were used as liver donors and Brown-Norway rats as recipients. The recipient rats were intravenously injected with anti 4-1BBL mAB or isotype control antibody. Groups were monitored for graft survival after transplantation. Plasma chemistry, including aspartate transaminase （AST）, alanine aminotransferase （ALT）, and bilirubin （BIL）, was assayed. The concentrations of interleukin （IL）-2, IL-10 and interferon （IFN）-γ in plasma were also measured by enzyme-linked immunosorbent assay. Allograft histology images were collected under light microscope and electron microscope. Results Isotype antibody treated recipients exhibited elevated plasma levels of liver injury markers including AST, ALT and BIL, progressive portal and venous inflammation and cellular infiltration of the liver ailografts, and a mean graft survival time （MST） of 10.9 days. Administration of anti 4-1BBL mAB resulted in a decrease in plasma levels of liver injury markers and the concentrations of IL-2, IL-10 and IFN-γ. The histological grade of rejection on day 7 decreased and MST （17.3 days） increased substantially. Conclusions These results demonstrate that attenuation of acute rejection follows the blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody and strongly suggest it is a promising strategy to prevent progression of graft rejection by suppressing T cell-mediated immunity.展开更多
Objective To investigate whether the impairment of grafted livers after transplantation was induced by the same inflammatory cells both in cold and warm ischemia. Methods Male SD rats were divided into two groups at r...Objective To investigate whether the impairment of grafted livers after transplantation was induced by the same inflammatory cells both in cold and warm ischemia. Methods Male SD rats were divided into two groups at random,24 donor livers in each group were stored in Ringers solution at 4℃ for 120min or 240min of transplantation for blood sample and tissue specimen collection. Results Along with the prolongation of cold and warm ischemia time,the serum ALT,AST and LDH level increased gradually after transplantation.Under light microscopy,some hepatocytes presented necrosis after 3h and 6h of transplantation in cold ischemia,and neutrophilic infiltration in sinusoids were evident.Also,a large number of hepatocytes were necrotic 3h or 6h after transplantation in warm ischemia from NHBDs,and lymphocytic infiltration was evident in the sinusoids.The findings in electron microscopy was as the same as those of light microscopy,and the cells which infiltrated the sinusoids in warm ischemia were identified as T lymphocytes. Conclusion The impairment of grafted livers after transplantation appeared to be induced by two different kinds of inflammatory cells in cold and warm ischemia,that is,neutrophils mediated the cold ischemia-reperfusion,and T lymphocytes mediated the warm ischemia-reperfusion from NHBDs,but these findings are to be comfirmed in further investigations.展开更多
Objective To explore the survival time,pathological change and liver regeneration in different kinds of reduced-size liver transplantation in rats using steatotic grafts. Methods Macrovesicular and microvesicular st...Objective To explore the survival time,pathological change and liver regeneration in different kinds of reduced-size liver transplantation in rats using steatotic grafts. Methods Macrovesicular and microvesicular steatotic rat liver models were established by feeding rats with a diet consisting of 79% standard chow,20% lard and 1% cholesterol for different time periods. With modified two cuff vascular anastomoses and end-to-end sutures on the bile duct,reduced-size orthotopic rat liver transplantations were performed in an attempt to explore the ratio of graft weight to recipient body weight,recipient original liver weight and histological and electron-microscopic findings in comparison with whole rat liver transplantations. Results A one-week survival rates for the rats undergoing whole liver transplantation,and those in the 70% reduced-size orthotopic liver transplantation (ROLT) group,the 60%ROLT group and the 50%ROLT group (grade Ⅰ macrosteatotic grafts) were 91.67%,75%,75% and 25%. A 2-week survival rate was 83.33%,75%,58.33% and 0 respectively. And their graft recipient body weight (GRBW) values SD were 3.56%±0.36%,2.53%±0.15%,2.28%±0.12% and 1.83%±0.16%,respectively. In grade Ⅱ macrosteatotic grafts,the one-week survival rate for those undergoiong whole liver transplantation and those in the 70% ROLT group was 83.33% and 25%. In the microsteatosis grafts for whole liver transplantation,70% ROLT,60% ROLT and 50% ROLT,the one-week survival rate was 83.33%,75%,75% and 33.33%; and the 2-week survival rate was 75%,66.67%,66.67% and 0,respectively. The survival rate of the 50% ROLT group using grade Ⅰ macrosteatotic grafts or grafts mainly with microsteatosis was significantly different from that of other groups. While using macrosteatotic grafts in grade Ⅱ, the 1-week survival rate of the 70% ROLT group was very poor. Pathological findings after operation included liver regeneration and portal space with mild lymphocyte infiltration. Improvement in steatosis and dilation of the central vein and sinusoids was observed in some rats.Conclusions In the successful and long-term survival of rat reduced-size liver transplantation using grade Ⅰmacrosteatotic grafts or grafts with microsteatosis,the GRBW values should be over 2.28%±0.12%,and the value of graft-recipient liver weight should be over 60%. Steatotic livers in grade Ⅱ should not be used as grafts in ROLT. Steatosis was improved and even totally cured in some long-term survival rats.展开更多
In order to understand the allograft rejection in orthotopic liver transplantation (OLT), an allograft re- jection rat model was established and studied by proteomic approach. The protein expression profiles of liver ...In order to understand the allograft rejection in orthotopic liver transplantation (OLT), an allograft re- jection rat model was established and studied by proteomic approach. The protein expression profiles of liver tissues were acquired by fluorescence two-dimensional difference gel electrophoresis (2D DIGE) that incorporated a pooled internal standard and reverse fluorescent labeling method. The expression levels of 27 protein spots showed significant changes in acute rejection rats. Among these spots, 19 were identified with peptide mass fingerprinting using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) after tryptic in-gel digestion. The results of the present paper could be helpful for our better understanding of allograft rejection in organ transplantation.展开更多
基金Supported by Grants from Jiangsu Provincial Government,China,No. ZX200605
文摘AIM:To investigate the impact of different time points of secondary warm ischemia on bile duct in a rat autologous liver transplantation model with external bile drainage.METHODS:One hundred and thirty-six male inbred SD rats were randomly assigned to one of four groups(Ⅰ-Ⅳ) according to the secondary warm ischemia time of 0,10,20 and 40 min.A rat model of autologous liver transplantation with continuous external biliary drainage under ether anesthesia was established.Ten rats in each group were used to evaluate the one-week survival rate.At 6 h,24 h,3 d and 7 d after reperfusion of the hepatic artery,6 rats were killed in each group to collect the blood sample via the infrahepatic vena cava and the median lobe of liver for assay.Warm ischemia time of liver,cold perfusion time,anhepaticphase,operative duration for biliary external drainage and survival rates in the four groups were analyzed for the establishment of models.RESULTS:No significant difference was shown in warm ischemia time,anhepatic phase and operative duration for biliary external drainage among the four groups.Five of the 40 rats in this study evaluated for the one-week survival rate died,including three deaths of severe pulmonary infection in group Ⅳ.A significant decrease of one-week survival rate in group Ⅳ was noted compared with the other three groups.With the prolongation of the biliary warm ischemia time,the indexes of the liver function assessment were significantly elevated,and biliary epithelial cell apoptosis index also increased.Pathological examinations showed significantly aggravated inflammation in the portal area and bile duct epithelial cell injury with the prolonged secondary warm ischemia time.Microthrombi were found in the micrangium around the biliary tract in some sections from groups Ⅲ and Ⅳ.CONCLUSION:The relationship between secondary warm ischemia time and the bile duct injury degree is time-dependent,and 20 min of secondary warm ischemia time is feasible for the study of bile duct injury.
文摘Background Blocking the 4-1BB/4-1BB ligand （4-1BBL） signal may modulate the secretion of Th1/Th2 cytokines and prolong the survival of the grafts, which play a key role in organ transplantation tolerance. The aim of this study was to investigate the role of blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody （mAB） in acute rejection of rat orthotopic liver transplantation. Methods The orthotopic liver transplantation model was set up, while male Lewis rats were used as liver donors and Brown-Norway rats as recipients. The recipient rats were intravenously injected with anti 4-1BBL mAB or isotype control antibody. Groups were monitored for graft survival after transplantation. Plasma chemistry, including aspartate transaminase （AST）, alanine aminotransferase （ALT）, and bilirubin （BIL）, was assayed. The concentrations of interleukin （IL）-2, IL-10 and interferon （IFN）-γ in plasma were also measured by enzyme-linked immunosorbent assay. Allograft histology images were collected under light microscope and electron microscope. Results Isotype antibody treated recipients exhibited elevated plasma levels of liver injury markers including AST, ALT and BIL, progressive portal and venous inflammation and cellular infiltration of the liver ailografts, and a mean graft survival time （MST） of 10.9 days. Administration of anti 4-1BBL mAB resulted in a decrease in plasma levels of liver injury markers and the concentrations of IL-2, IL-10 and IFN-γ. The histological grade of rejection on day 7 decreased and MST （17.3 days） increased substantially. Conclusions These results demonstrate that attenuation of acute rejection follows the blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody and strongly suggest it is a promising strategy to prevent progression of graft rejection by suppressing T cell-mediated immunity.
文摘Objective To investigate whether the impairment of grafted livers after transplantation was induced by the same inflammatory cells both in cold and warm ischemia. Methods Male SD rats were divided into two groups at random,24 donor livers in each group were stored in Ringers solution at 4℃ for 120min or 240min of transplantation for blood sample and tissue specimen collection. Results Along with the prolongation of cold and warm ischemia time,the serum ALT,AST and LDH level increased gradually after transplantation.Under light microscopy,some hepatocytes presented necrosis after 3h and 6h of transplantation in cold ischemia,and neutrophilic infiltration in sinusoids were evident.Also,a large number of hepatocytes were necrotic 3h or 6h after transplantation in warm ischemia from NHBDs,and lymphocytic infiltration was evident in the sinusoids.The findings in electron microscopy was as the same as those of light microscopy,and the cells which infiltrated the sinusoids in warm ischemia were identified as T lymphocytes. Conclusion The impairment of grafted livers after transplantation appeared to be induced by two different kinds of inflammatory cells in cold and warm ischemia,that is,neutrophils mediated the cold ischemia-reperfusion,and T lymphocytes mediated the warm ischemia-reperfusion from NHBDs,but these findings are to be comfirmed in further investigations.
文摘Objective To explore the survival time,pathological change and liver regeneration in different kinds of reduced-size liver transplantation in rats using steatotic grafts. Methods Macrovesicular and microvesicular steatotic rat liver models were established by feeding rats with a diet consisting of 79% standard chow,20% lard and 1% cholesterol for different time periods. With modified two cuff vascular anastomoses and end-to-end sutures on the bile duct,reduced-size orthotopic rat liver transplantations were performed in an attempt to explore the ratio of graft weight to recipient body weight,recipient original liver weight and histological and electron-microscopic findings in comparison with whole rat liver transplantations. Results A one-week survival rates for the rats undergoing whole liver transplantation,and those in the 70% reduced-size orthotopic liver transplantation (ROLT) group,the 60%ROLT group and the 50%ROLT group (grade Ⅰ macrosteatotic grafts) were 91.67%,75%,75% and 25%. A 2-week survival rate was 83.33%,75%,58.33% and 0 respectively. And their graft recipient body weight (GRBW) values SD were 3.56%±0.36%,2.53%±0.15%,2.28%±0.12% and 1.83%±0.16%,respectively. In grade Ⅱ macrosteatotic grafts,the one-week survival rate for those undergoiong whole liver transplantation and those in the 70% ROLT group was 83.33% and 25%. In the microsteatosis grafts for whole liver transplantation,70% ROLT,60% ROLT and 50% ROLT,the one-week survival rate was 83.33%,75%,75% and 33.33%; and the 2-week survival rate was 75%,66.67%,66.67% and 0,respectively. The survival rate of the 50% ROLT group using grade Ⅰ macrosteatotic grafts or grafts mainly with microsteatosis was significantly different from that of other groups. While using macrosteatotic grafts in grade Ⅱ, the 1-week survival rate of the 70% ROLT group was very poor. Pathological findings after operation included liver regeneration and portal space with mild lymphocyte infiltration. Improvement in steatosis and dilation of the central vein and sinusoids was observed in some rats.Conclusions In the successful and long-term survival of rat reduced-size liver transplantation using grade Ⅰmacrosteatotic grafts or grafts with microsteatosis,the GRBW values should be over 2.28%±0.12%,and the value of graft-recipient liver weight should be over 60%. Steatotic livers in grade Ⅱ should not be used as grafts in ROLT. Steatosis was improved and even totally cured in some long-term survival rats.
基金the Key Basic Research and Development Program of China (Grant No. 2003CB515506)in part by the Chinese Human Liver Proteome Project (CNHLPP) (Grant No. 2004BA711A18)
文摘In order to understand the allograft rejection in orthotopic liver transplantation (OLT), an allograft re- jection rat model was established and studied by proteomic approach. The protein expression profiles of liver tissues were acquired by fluorescence two-dimensional difference gel electrophoresis (2D DIGE) that incorporated a pooled internal standard and reverse fluorescent labeling method. The expression levels of 27 protein spots showed significant changes in acute rejection rats. Among these spots, 19 were identified with peptide mass fingerprinting using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) after tryptic in-gel digestion. The results of the present paper could be helpful for our better understanding of allograft rejection in organ transplantation.
