Hepatic ischemia/reperfusion injury (HIRI) exists in lots of process of clinical pathology and operation. Apoptosis is an active process controlled by some gene and factors such as Fas/FasL, Caspases and Bcl-2 families. More and more studies suggest that HIRI is associated with apoptosis. This article summarized the mechanisms and gene modulation of apoptosis during HIRI and the significance of suppressing apoptosis in preventing HIRI.

Objective To investigate the surgical technique of establishing a reliable rat model of orthotopic liver transplantation and prevention of operational complication. Methods The model was established with modified cuff technique.The donor was perfused through the abdominal aorta with 20ml cold perfusate.The anastomosis of the suprahepatic vena cave(VC)was sutured end-to-end with 8-0 nylon line,and the continuity of infrahepatic VC and portal vein (PV)were established by means of cuff method respectively.The bile duct anastomosis was performed by internal stent. Results Three hundred and sixty case of rat orthotopic liver transplantation were performed and the successful rate of the mode was 91.3%.In non-intervention group,the survival rat was satisfactory,with a 86.5%of 1 week survival and a 80.7% of long-term survival (longer than 3 months). Conclusions The sophisticated microsurgical technique and the delicate surgical manipulation is the prerequisite of preventing operational complication,and shortening the anhepatic phase of recipient as soon as possible is the key to animal survival.

Objective:To investigate the techniques of establishment of abdominal heart transplantation model in rats and modify the procedure to make it more stable and improve the success rate.Methods: Heterotopic abdominal heart transplantations of Wistar and SD rats were performed by using Ono's method with modification of preparation of recipients,donor cardiectomy and donor transplantations.Results: One hundred heart transplantations were performed,among which 90 were successful.The success rate was 90%.Conclusion: This modified Ono's method can shorten the ischemia time and recipient abdominal aorta occlusion time,make vessel anastomosis easier and increase the success rate of operation.It is a stable and reliable model of heart transplantations in rats.

OBJECTIVE:To explore the repairing effect and mechanism of Huazhuo jiedu yukui formula on the mucosal dam-age of rats with ulcerative colitis (UC). METHODS:72 rats were randomly divided into blank group,model group,mesalamine group(positive control,0.3 g/kg),Huazhuo jiedu yukui formula low-dose,medium-dose,high-dose groups(5,10,20 g/kg),12 in each group. Except for blank group,other groups were induced UC model. After 4 d of modeling,rats in administration groups received related medicines,ig,blank group and model group received normal saline,ig,once a day,for 2 weeks. After administra-tion,disease activity index(DAI)and mucosal damage index(CMDI)of rats were observed;IL-8,TNF-α contents in serum and cyclooxygenase 2 (COX-2) expression in colon tissue were detected. RESULTS:Compared with blank group,DAI and CMDI scores in model group were increased;IL-8,TNF-α contents in serum were increased;COX-2 expression in colon tissue was en-hanced,with statistical significances (P<0.05). Compared with model group,DAI and CMDI scores in model group were de-creased;IL-8,TNF-α contents in serum were reduced;COX-2 expression in colon tissue was weakened,with statistical signifi-cances (P<0.05). Compared with mesalamine group, the above-mentioned indicators in Huazhuo jiedu yukui formula medi-um-dose,high-dose groups changed more obviously (P<0.05),and effects of Huazhuo jiedu yukui formula showed significant dose-effect relationship. CONCLUSIONS:Huazhuo jiedu yukui formula can improve disease activity status of UC rats and reduce colon mucosal damage,and the effects of medium-dose and high-dose Huazhuo jiedu yukui formula were better than mesalamine, which may be related to reducing inflammatory reaction and inhibiting COX-2 expression in colon tissue.

Objective To observe recipient survival time and liver graft regeneration in an orthotopic liver transplantation model using IL-6 knockout mice as a donor.Methods A model of liver transplantation in C57BL/6 WT and IL-6 KO mice with C57BL/6 background was established.Thirty eight mice were divided into three groups:C57BL/6 wild type→C57BL/6 wild type control group(n=10),IL-6 KO→IL-6 KO group(n=14)and IL-6 KO→C57BL/6 WT group(n=14).Hepatocyte replication with BrdU uptake after liver transplantation was examined by immunohistochemistry.Resuits The survival time in the control group was＞16 days.and that was 2 days and 1.6 days in the IL-6 KO→IL-6 KO and IL-6KO→C57BL/6 WT,respectively.The difierence in survival time in all three groups was statistically significant(F=190.09,P＜0.01).The liver grafts in control group showed minimal injury and no necrosis and mild increase of BrdU uptake at 48 hours.Patchy areas of necrosis and hepatocyte ballooning were observed in the two groups using IL-6 KO mice as donors,there was minimum increase in BrdU uptake at 48 hours post transplantation.Conclusion IL-6 KO liver grafts fail to regenerate after liver transplantation.IL-6 is an important factor in liver regerative response.

Objective To study the molecular mechanism involved in early liver regeneration following cold ischemia after rat orthotopic liver transplantation(OLT). Methods Syngeneic(Lewis to Lewis)rat OLT with hepatic artery reconstruction was performed.Graft survival and liver regeneration following 1 h and 16 h cold ischemic injury were evaluated.Specimen were collected at predetermined intervals from 0,1,2,4,16,24,48,72 h post-reperfusion.The patterns of IL-6 and TNF-α expression,STAT3 activation,and upregulation of immediately early genes(IEGs)including c-fos,c-myc,and c-jun were determined in liver grafts with 1 h and 16 h cold ischemia times.Quantitative analysis of IL-6 at 2 h after reperfusion was performed for grafts preserved for 1 h and 16 h.Progression of hepatocyte replication was confirmed by bromodeoxyuridine(BrdU)immunohistochemistry.Positively BrdU-stained nuclei were quantitated and analyzed between the two groups at 48 h after reperfusion. Results Survival rate in the two groups was 100%.Compared to 1 h cold ischemia group,IL-6 and TNF-α expression and STAT3 activation in 16 h cold ischemia group were markedly increased.Extensive hepatocyte replication was present.Upregulation of immediate early genes of c-fos,c-myc,and c-jun was also observed.Grafts with 1 h ischemic iniury had lower levels of IL-6 than that when grafts suffering from ischemia for 16 h(t=27.7,P＜0.05).Number of positively stained nuclei in 16 h group were more than that in 1 h group at 48 h after transplantation(t=13.4,P＜0.05). Conclusions Severe cold ischemia initiated liver regeneration after rat OLT.TNF-α and IL-6 are important factors in early liver regeneration.IL-6/STA3 pathway in liver regeneration is an important part in the recovery of grafts following cold ischemia injury.

Objective To explore the optimal timing of liver transplantation for acute liver failure,and analyze relative risk factors affecting patients' prognosis.Methods We retrospectively analyze perioperative clinical data of 50 patients suffering from acute liver failure who underwent liver transplantation between March 2005 and June 2010 in Organ Transplantation Center,the First Affiliated Hospital of Sun Yat-sen University.Patients were divided into survival group and death group.Clinical data before operation were collected and analyzed by univariate and multivariate analysis,respectively.Risk factors influencing the perioperative mortality were selected and were used to direct the operation time.Results There were 50 patients who underwent liver transplantation and 11 cases died during perioperative phase.Five patients died of respiratory infection,three of acute renal failure,one of digestive tract hemorrhage,one of primary allograft failure and one of multiple organ failure.The perioperative,one-year and three-year survival rate were 78％,74％ and 72％ respectively.In the univariate analysis,age,bilirubin level,prothrombin time,creatinine level,grade of encephalopathy and MELD score were found to be significantly different between survival group and death group.The multivariate analysis revealed that age≥65 y,INR ≥3.5,creatinine level ≥ 2.5 mg/dl and MELD score ≥ 40 were independent risk factors affecting survival.Conclusions Liver transplantation is an effective treatment of choice for acute liver failure.Appropriate timing of transplantation helps achieve a high survival rate.

Objective To investigate the histological and ultrastructural characteristics of liver graft after different warm ischemia time (WIT) in rats. Methods According to WIT, rats were randomized into 7 groups, with WIT of 0, 10, 15, 20, 30, 45, 60 minutes respectively. All specimens were investigated by light, electron microscopy, and histochemistry stain. 6, 24, and 48 hours after orthotopic liver transplantation(OLTx) ,the graft morphology was observed. Results The donor liver from non-heart-beating donors (NHBO) underwent ischemia injury both in the warm ischemia period and in the reperfusion period. Morphological changes were positively related to warm ischemia time in a time-dependent manner during the reperfusion period. There was a histocytic degeneration of different degree within 30 minutes warm ischemia. Although becoming more severe with the prolongation of warm ischemia time within this period, there was no obvious hepatocyte necrosis in any specimens. In WIT 45 min group, small focal necrosis occurred which was found in central area of hepatic lobule first. In 60 min group, patchy or diffused necrosis was observed and the area was gradually extended, while hepatic sinusoid endothelial cell obviously swelled to be bleb or balloonlike, hepatic sinusoid was obstructed and microcirculation was in disorder. Conclusion Rat liver graft undergoing warm ischemia injury is on the reversible stage within 30 minutes warm ischemia time by histological, histochemistry and ultrastructural dynamic observation. 45 min is a critical point of rat liver graft to endure warm ischemia injury, and when WIT was over 60 min, the damage is irreversible.

OBJECTIVETo explore the diagnosis, treatment and long-term outcome of late acute rejection (LAR) following adult orthotropic liver transplantation (OLT).

METHODSA total of 398 consecutive adult patients who underwent OLT in Organ Transplant Center, the First Affiliated Hospital, Sun Yat-sen University between January 2007 and December 2012 were reviewed retrospectively. There were 48 patients (12. 1%) developed to LAR, including 43 male patients and 5 female patients, with an average age of (52 ± 13) years(18 - 70 years). The mean body mass index was (22.1 ± 4. 5) kg/m2 (15. 4 - 30. 4 kg/m2). The indications of the liver transplantation recipients included 16 cases of end-staged liver cirrhosis after hepatitis B or C(33. 3%), 14 cases with severe hepatitis (29. 2%), 9 cases of primary liver cancer(18. 5%), 5 cases of alcoholic liver cirrhosis (10. 4%), 1 case with autoimmune liver disease (2. 1%) , the other 3 cases (6. 3%). They were followed up by outpatient service, telephone and other means. Survival curves were generated with the Kaplan-Meier method and Cox proportional hazards modeling was used for predictors of mortality. Statistically significant variables found by single factor regression analysis were put into the Cox proportional hazards regression model of multivariate analysis.

RESULTSThe time-to-event was 23. 6 months after OLT which were more common in the first year to the third year post-transplant (26/48,52. 4%). Thirty-five cases were assessed as mild, 11 cases were assessed as moderate, and 2 cases were assessed as severe ,based on the Banff schema. After adjustment to the immunosuppressive regimen, the overall recovery rate reached to 81. 3%. The rate of steroid-resistant acute rejection was 11. 8% (4/34). Inadequate immunosuppression and steroid pulsation were two independent risk factors affecting the prognosis of LAR (P = 0. 008, P = 0. 003, respectively).

CONCLUSIONSLAR is an uncommon complication after OLT. Inadequate immunosuppression and steroid pulsation are the major risk factors for prognosis of LAR. Improving patient compliance and strengthening blood concentration surveillance can increase the patient survival.

Acute Disease ; Adolescent ; Adult ; Aged ; Female ; Graft Rejection ; diagnosis ; mortality ; therapy ; Hepatitis B ; Humans ; Immunosuppression ; Liver Cirrhosis ; Liver Neoplasms ; Liver Transplantation ; mortality ; Male ; Middle Aged ; Multivariate Analysis ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Risk Factors ; Steroids ; Young Adult

Objective Sirolimus is a new, potent immunosuppreasant considered to be nonnephrotoxic. There is limited experience with the use of sirolimus in liver transplant recipients. This study was to investigate the clinical experience of conversion from tacrolimus-based to sirolimus-based immunosuppression in liver transplant recipients. Patients switched to cyclosporine-based immunosuppression during the same period were also enrolled as controls. Methods This retrospective study examined liver transplant recipients who had been switched from tacrelimus-based to sirolimus-based or cyelosporine-based immunosuppressive therapy between January 2004 and January 2008 in the First Affiliated Hospital of Sun Yat-sen University. Patients were divided into 2 groups: those switched to sirolimus-based immunosuppression (group A; n=32); and those switched to cyclosporine-based immunosuppression (group B; n=15). Results The rate of successful conversion was 34.5% in group A (10/32) compared with 45.5% in group B (7/15); this difference was not statistically significant (P>0.05). After conversion, renal function in patients in group A remained normal, while the renal function in patients in group B become abnormal 4 months after conversion (P<0.05). In group A, some simlimus-associated adverse effects occurred but were mild and easy to control. Conclusion Sirolimus can be used safely in place of tacrolimus in liver transplant recipients.