Objective To explore the mechanism of hepatic autophagy inhibition induced by ischemia-reperfusion injury in the aging liver.Methods The healthy male Lewis rats aged 3 months (3M) and 24 months (24M) were selected,and then were randomly divided into 3M IRI group,3M sham operation group,24M IRI group,24M sham operation group.In the experimental group,noninvasive vascular clamp was used to clamp the left and middle hepatic lobes (about 70o％ hepatic ischemia).The liver was subjected to ischemia at 37 0.5℃ for 30min and reperfusion for 6h.The hepatic duodenal ligament was dissected only by sham operation.The serum levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured at 6 h after operation.Liver tissues of each group were examined by liver pathology and the number of autophagosome of LC3B in liver tissue of each group was observed under confocal microscopy.The changes of autophagyrelated protein were analyzed by Western blotting.Results The levels of serum ALT and AST in 24M IRI group were significantly higher than those in 3M IRI group,the difference was statistically significant (P＜0.05).Pathological analysis showed that 3M IRI group showed spotty necrosis,the 24M IRI group showed massive necrosis and the infiltration of the inflammatory cells;Confocal microscopy showed that the number of autophagosome in the liver tissue of the 24M sham group was slightly lower than that of the 3M sham operation group and the number of autophagosome in the 24M IRI group was significantly lower than that in the 3M IRI group (P＜0.05).The levels of autophagyrelated proteins (Beclin1 and ATG4B protein) in 24M IRI group were significantly down-regulated compare to 3M IRI group (P＜0.05).Conclusion The ischemia-reperfusion injury of liver in aged rats inhibits autophagy,and its mechanism may be related to the decrease of autophagy-related protein level in hepatic ischemia-reperfusion injury.

Objective To explore the cause and treatment strategy of chronic graft rejection characteristic of jaundice in patients with liver transplantation. MethodsPrimary disease, immunosuppressive protocol were reviewed in nine cases surviving more than 1 year after liver transplantation. The pathology examination, choledochoscopy and ultrasound examination were performed. The dosage of immunosuppressive agent was adjusted. Four cases underwent operation. Results Chronic rejection was well controlled in 6 out of the 9 cases with total bilirubin level decreased from 200 ?mol/L to less than 100 ?mol/L. Bile flocculus and bile slush became less obvious or disappeared after flushing and dilataltion of the common bile duct. Three cases died in spite of aggressive therapy. ConclusionsInsufficient immunosuppressive strength is one of the most important causes for chronic rejection. Immunological injury results in hepatocellular damage, bile slush, inflamed thick bile duct.

Objective To investigate the effect of down-regulated Blimp1 gene expression on differenetiation of bone marrow cells into dendritic cells (DCs).Methods Blimp1-shRNA was constructed and then loaded into lentivirus vector as lenti-blimp1-shRNA.Bone marrow cells from Balb/c mice were induced differentiation to DCs in an 8-day cell culture system with GM-CSF/IL-4 incubation and LPS stimulation at day 7.The cells were divided into groups as empty control (no treatment),lenti-control (transfected by lentivirus empty vector at day 1),and lenti-Blimp (transfected by lenti-blimp1-shRNA at day 1).The transfection efficiency was evaluated by GFP fluorescence for one week.The morphology and growth curve were analyzed.Real-time PT-PCR and Western blotting were used to evaluate mRNA and protein expression of Blimp1.At day 8,CD11 c and CD86/MHC-Ⅱ were quatitified using flow cytometry.Results GFP fluorescence emerged 3 days after transfection and was continuously expressed.Classic DC morphology was shown in no treatment cells,while damaged morphology presented in the cells with lentivirus transfection.The empty control cells proliferated from day 3,peaked as (2.45 ± 0.26) 106/well at day 4,and kept at (2.27 ± 0.19) 106/ well at day 8,The cells receiving lentivirus presented (1.69 ± 0.39) 106/well.The expression of Blimp1 mRNA and protein in the lenti-Blimp1 group was 76％/1％ and 1.0％/74.0％ of the empty control group.At day 8,CD11c,CD86 and MHC-Ⅱ expression in the empty control group was (69.2 ±5.0)％,(51.1± 4.9) ％ and (56.3 ± 7.3) ％,while (68.6±5.9)％,(49.5±4.3)％ and (69.4±4.5)％ in the lenti-control group,and (72.8 ± 5.5)％,(50.2 ± 6.0)％ and (46.5 ± 5.7)％ in the lenti Blimp1 group.Conclusion Lentivirus-mediated Blimp1-shRNA gene therapy modulates blimp1 expression of DC precursors.Down-regulation of Blimp1 fails to interrupt the differentiation of DCs but inhibits the maturation.

Objective To determine the long-term results after combined pancreas-kidney transplantation at a single-center institution.Methods Fifty-three consecutive patients with insulin-dependent diabetes mellitus and end-stage nephropathy were followed up for more than three years after combined pancreas-kidney transplantation. Immunosuppressive protocol consisted of tacrolimus ( TAC ),mycophenolate mofetil (MMF),and steroids,and antithymocyte globulin or anti-CD25 receptor mAb.The impact of different risk factors was analyzed on long term patient and graft survival.Results The 3-,5- and 8-year survival rate in recipients was 90.1％,89.1 ％ and 80.0％,respectively.The 3-,5- and 8-year survival rate of pancreas grafts was 84.9％,84.8％ and 60.0％,and that of kidney grafts was 83.0％,82.6％ and 53.3％,respectively.Principal causes of death were Infection (n =4),renal failure (n =2),cardiovascular events (n =1 ),and cerebrovascular accident (n =1 ).Graft failure for the pancreas was caused by death with a functioning graft (n =6),rejection (n =2),thrombosis (n =1 ) and pancreatitis (n =1 ).Graft failure for the kidney was due to rejection (n =9),and death with a functioning graft (n =9).Conclusion This series representing the largest experience with long-term follow up in China confirms an excellent long-term survival.Infection,rejection and surgical complication were the major risk factors leading to deaths and graft loss.

Objective To evaluate the relevant causes of neurologic complications following liver transplantation.Methods 155 adult patients (131 males, 24 females) who received liver transplantation for the first time at Tongji Hospital between January 2005 and September 2009 were identified.Case notes were reviewed and demographic data, details of the liver disease, neurologic complications, MELD score and discharge information were recorded.Results Neurologic complications occurred following 36 transplants (23.2 %), The complications included mental symptoms in 15 cases (41.7 %), disorder of consciousness and action in 9 cases (25 %), and coma in 12 cases (33.3 %).Twelve percent patients with liver cancer experienced a neurologic complication, which was lower than for other transplant indications, like acute and chronic hepatic failure because of HBV infection (33.3 %, P＜0.01), inborn/metabolic disease (40 %, P＜0.05), and HCV Infection (25 %, P = 0.36).Patients who experienced a neurologic problem had significantly higher MELD score (for non-cancer patients:22.93 ± 8.21; for cancer patients:17 ± 5.4) than the other Patients (for non-cancer patients:18.33 + 8.47, P＜0.05; for cancer patients:13 ±3.4, P＜0.01).The rate of infection (36.1 %) and mortality (30.5 %) were significantly higher in patients with neurologic complications (P＜0.01).The levels of ALT, TBil, ALB, PT and the concentrations of serum sodium and chlorine had no impact on neurologic complications.Conclusion Neurologic complications are common in liver transplant recipients.These complications are related to primary disease and liver function before the operation, and increase the rate of infection and mortality.

objective From April 1994 to April 1997, the authors anatomized and studied 40 cadaveric Livers, Which included the character, Length and diameter of the vessels and bile ducts of the first, second and third hepatic portis. This article put the emphasis on the anatomic structures and its clinical applilation. The results of the 40 cadaveric liver anatomy will play important roles on the resection and transplantation of the liver, especially the Piggy back Liver transplantation.

Objective To explore the prophylaxis and treatment strategies for psychic syndrome in patients after piggyback liver transplantation.Methods The data on the etiology,treatment outcome and prognosis of psychic syndrome occurring in 45 of 235 patients who had piggyback liver transplantation were retrospectively analyzed.Results The incidence of psychic syndrome complication was 19.1%(45/235),22 cases presented as mania(48.9%),5 cases as tristimania(11.1%),3 cases as hallucinosis(6.7%),1 case as suicidal tendency(2.2%),1 case as metamorphopsia(2.2%),8 cases as angst insomnia(17.8%),2 cases as maladjusted disturbance(4.4%),3 cases as affective disturbance(6.7%),and the majority presented as delirious alienation.All the cases were cured,except 1 case of coma,who was confirmed by CT to have intracranial hemorrage,died after failure of resuscitation.Conclusions The incidence of psychic syndrome in patients after piggyback liver transplantation is relatively high.However,most cases have mild symptoms and the prognosis is fine.When the patients have psychogenic symptoms,the prognosis of patients can be improved by some symptomatic treatment strategies directed to their different clinical manifestations.

Objective To investigate the clinical characteristics and strategies of early stage antibody-mediated rejection after renal transplantation.Method The clinical data of early stage AMR of 3 cases of renal transplantation,and 1 case of pancreas transplantation after renal transplantation were retrospectively analyzed.(1) The case 1 was diagnosed as having early severe acute AMR.Serum creatinine was increased,urine volume rapidly reduced,the blood flow of transplanted kidney reduced on the postoperative day 8;the positive rate of panel reactive antibody (PRA) class Ⅰ and Ⅱ was 74.6％,and 2.7％ respectively on the postoperative day 12.Biopsy showed widely ischemia and local bleeding in transplanted kidney and DSA showed anti-B62 mean fluorescence intensity (MFI) increased to 6800 on the postoperative day 14.(2) The case 2 was diagnosed as having early mild acute AMR.The positive rate of PRA class [and Ⅱ was 65.6％ and 78.9％ respectively.DSA Ⅰ was positive,anti A11 MFI was 3059,and DSA Ⅱ was negative on the postoperative day 13.Biopsy showed mild ischemia reperfusion injury in transplanted kidney on the postoperative day 21.(3) The case 3 was diagnosed as having early severe chronic AMR,and the recipient received pancreas transplantation 1 year after kidney transplantation.Eight months after pancreas transplantation,DSA for pancreas donor was detectable,anti A2 MFI was 7514,anti B46 MFI was 3 159 and anti DQ7 MFI was 1 503.(4) The case 4 was diagnosed as having early mixed rejection.Serum creatinine was elevated on the postoperative day 8;PRA testing showed that the positive rate of class Ⅰ and Ⅱ was 3％ and 70％ respectively,DSA was positive,and anti DR16 MFI was 15 170 on the postoperative day 14;transplanted kidney biopsy showed acute mixed rejection on the postoperative day 16.Result Case 1 and case 3 were not diagnosed and treated in time and graft loss developed.Case 2 and case 4 were functionally recovered after combined treatment of plasmapheresis,IVIG and bortezomib.Conclusion Diagnosis of antibody-mediated rejection is based on transplant graft dysfunction,positive DSA and graft biopsy.Early diagnosis,early treatment and combined therapy can improve the curative rate of AMR.

Objective To analyze the curative effect of simultaneous liver and kidney transplantation (SLKT) for patients with end-stage liver and kidney diseases and liver cirrhosis patients with hepatorenal syndrome.Methods All SLKTs (n=21) performed at our center from January 1999 to December 2010 were reviewed and SLKT outcomes were compared with those of kidney transplantation (KT) (n=609) and liver transplantation (LT) (n=133) performed during the same period.Results There were 3 deaths due to infection 2 weeks, 6 months and 5 years respectively after operation. One patient died due to multiple organ dysfunction syndrome 2 weeks after operation. One patient was dead 5 years after operation because of rejection. MELD level between SLKT and LT had no significant difference, but serum creatinine in SLKT group was significantly higher than in LT group (516.0±329.9 vs 111.4±138.1 mmol/L, P<0.01). The 1-year acute kidney rejection rate and rate of delayed graft function (DGF) of the kidney had no significant difference between SLKT group (0 vs 9.5 %) and KT group (6 % vs 17.3 %). There was no significant difference in one-, 3- and 5-year patient survival rate between SLKT group (87.7 %, 67.8 % and 67.8 %) and LT group (84.2 %, 73.5 % and 69.4 %).Conclusion SLKT is a safe and effective treatment for end-stage liver and kidney diseases.

Objective To investigate the effects of T. gondii soluble tachyzoite antigen (STAgs) on the survival time of mouse heart allograft and the possible mechanism. Methods The STAgs were prepared by pulverizing T. gondii tachyzoite with ultrasound on ice. Cervical heterotopic heart transplantations were done by using Balb/c mice as donors, and C57BL/6 mice as recipients.The recipients were classified randomly into three groups: syngeneic group, acute rejection group and STAgs-treated group. The recipients in acute rejection group and STAgs-treated group were injected subcutaneously with 0. 1 ml PBS and 0. 1 ml (5 μg) STAgs at the 4th day before transplantation respectively, and those in syngeneic group were not subjected to any treatment. The grafts were observed daily by cervical palpation, and the total cessation of cardiac contraction was defined as the endpoint. The heart allografts were harvested at the 7th day after transplantation for pathological examination and immunohistochemical staining for CD4+ T, CD8+ T. Results The recipients in syngeneic group were all alive at the 100th day after transplantation. The average survival time in acute rejection group and STAgs-treated group was (6.7± 0.5) days and (70.8± 3.5) days,respectively (P＜0.05). HE staining showed that the rejection on the 7th day after transplantation in syngeneic group, acute rejection group and STAgs-treated group was fallen into 0 degree, Ⅲ-Ⅳ degree and 0- Ⅰ degree, respectively. Immunohistochemical staining revealed that the CD4+ T and CD8+T were markedly down-regulated in STAgs-treated group as compared with those in acute rejection group. Conclusion T. gondii STAgs can significantly prolong the survival time of mouse heart allograft and inhibit the rejection probably by changing the ratio of TH1/TH2, or inhibiting the effect of dendritic cells by inducing the lipoxin A4.