Objective To investigate the effects of ischemic preconditioning on the cholesterol content and the activity of Na+-K+-ATPase of hepatocytes following cold preservation in rats.Methods Twenty-five rats were randomly divided into five groups,including control group (C),cold preservation group (Ⅰ),ischemic preconditioning group (ⅠP),atorvastatin (30 μmol/L) treatment group (A30),and atorvastatin (100 μmol/L) treatment group (A100).The cholesterol content and the activity of Na+ -K+ -ATPase were assessed.Results The cholesterol contents on the rat liver tissue cell membrane in the C group,Ⅰ group,ⅠP group,A30 group and A100 group were (310.4 ± 27.5),(187.7±13.1),(394.3±25.9),(201.8±14.6) and (122.6±7.7) nmol/mg protein,and activity of the Na+ -K+ -ATP enzyme was (46.55 ± 3.20),(27.4 ± 2.81),(52.71 ± 3.02),(30.67 ±2.78) and (19.64 ± 2.11) μmol Pi/hr mg protein,respectively (P＜0.05).There was no significant difference in the plasma membrane phospholipid content among the five groups (P＞0.05).Conclusion Reduction of cholesterol content and Na+ K+ -ATPase activity on the liver cytoplasmic membrane is one of the factors causing donor liver cold preservation injury,but ischemic preconditioning can significantly improve cell membrane Na+ -K+ -ATPase activity and increase cytoplasmic membrane cholesterol content. Use of atorvastatin statins can reduce cytoplasmic membrane cholesterol synthesis,and significantly decrease Na+ -K+ -ATPase activity,thereby alleviating the donor liver cold preservation injury.

Objective To evaluate the diagnosis and treatment of early-stage hepatic artery thrombosis(HAT) after adult liver transplantation.Methods387 consecutive adult patients who underwent liver transplantation from June 2007 to October 2010 by the same surgery team in the Transplant Center,First Affiliated Hospital of Sun Yat-sen University were retrospectively studied.Hepatic arterial blood flow was monitored by color Doppler ultrasound (DUS) daily during the first week after transplantation.Ultrasonic contrast or hepatic artery angiography was performed on recipients with suspected HAT.Results10 patients developed HAT on 7(2-18)d after operation.The incidence of HAT was 2.6％ (10/387).Interventional therapy was performed in 2 patients with one patient who received a stent because of hepatic artery stricture.Three patients underwent emergent hepatic artery revascularization combined with intra-arterial urokinase thrombolysis treatment.One developed a rethrombosis and died.The remaining 2 patients received re-transplantation.Three patients died of liver failure and severe infection.The mortality rate was 40％ (10/387).ConclusionsIt is essential to diagnoses HAT by monitoring the artery flow by Doppler ultrasound screening in the early period after operation.Interventional therapy,emergent hepatic artery revascularization and re-transplantation are effective rescue treatments.Prevention of HAT is most important.

Objective To evaluate the efficacy of liver grafts with warm ischemia and with different cold preservation time in liver transplantation.Methods The clinical data of 154 patients who received liver transplantation at the First Affiliated Hospital of Sun Yat-sen University from January 2006 to December 2007 were retrospectively analyzed.The warm ischemia time of the liver grafts obtained from the non-heart-beating donors was within 10 minutes.According to cold perservation time of the liver grafts,patients were divided into 3 groups:the cold preservation time of the liver grafts was within 8 hours,8-12 hours and above 12 hours in group I(n=58),group Ⅱ(n=62)and group Ⅲ(n=34),respectively.The peak level of alanine aminotransferase(ALT),primary graft dysfunction(PGD)after liver transplantation,acute rejection response,biliary complications,vessel complications,perioperative infections and the survival of liver grafts and recipients among the 3 groups were analyzed via chi-square test,t test and variance analysis.Results No PGD was detected in the 3 groups after liver transplantation.All patients were followed up for 8-32 months.The peak level of ALT,incidence of infection and biliary complication,survival of liver grafts and recipients were(482±357)U/L,12%(7/58),12%(7/58),86%(50/58)and 88%(51/58)in group Ⅰ,and were(1274±608)U/L,29%(10/34),26%(9/34),68%(23/34)and 71%(24/34)in group Ⅲ,with significant difference between the 2 groups(t=5.23,X~2=4.28,6.77,4.51,4.28,P<0.05).The peak level of ALT in group Ⅱ was(953±424)U/L,which was significant higher than(482±357)U/L in group Ⅰ(t=4.76,P＜0.05).Conclusions Liver grafts with a warm ischemia time shorter than 10 minutes could tolerate the injury caused by cold preservation with the maximum time of 12 hours.The incidences of biliary complications and postoperative infections are significantly increased and the survivals of liver grafts and recipients are decreased when the cold preservation time exceeds 12 hours.

BACKGROUND:Nowadays,liver grafts for transplantation are clinically sourced from non-heart-beating donors.Moreover,there is still no uniform determination of safe time limit points for non-heart-beating donor liver in warm ischemia and cold preservation.OBJECTIVE:To evaluate the application safety and curative effects of warm ischemia liver graft affected by varied cold ischemia time (CIT) in liver transplantation.DESIGN,TIME AND SETTING:A randomized controlled observation was performed in the Organ Transplantation Center,First Affiliated Hospital of Sun Yat-sen University between January 2006 and December 2007.PARTICIPANTS:154 cases who underwent non-heart-beating liver transplantation were included in this study.All liver graftsinvolved had a warm ischemia time(WIT) less than 10 minutes.METHODS:All cases were assigned into 3 groups according to CIT:group Ⅰ (CIT＜8 hours,n=58),group Ⅱ (CIT 8-12hours,n=62),and group Ⅲ (CIT 13-16 hours,n=34).The liver grafts were randomly allocated for the patients.Following liver transplantation,the same immunosuppression protocol was employed for each group.MAIN OUTCOME MEASURES:Following surgery,peak level of alanine aminotransferase (ALT),primary graft dysfunction (PGD) after liver transplantation,acute rejection response,biliary complications,vessel complications,perioperative infections,graft and recipient survival rate were compared among 3 groups.RESULTS:Follow-up time was 8-32 months.No PGD was detected in all 154 cases.Group Ⅱ showed postoperative ALT peak levels significantly higher than group Ⅰ (P＜ 0.05).There was no significant difference between groups Ⅰ and Ⅱin terms of acute cellular rejection,perioperative infection,biliary complication,vessel complication,graft survival rate,and recipient survival rate (P＞0.05).Compared with group Ⅰ,the group Ⅲ exhibited significantly increased postoperative ALT peak level,biliary complications,and perioperative infections,and significantly decreased graft and recipient survival rate (P＜0.05).CONCLUSION:Non-heat-beating-liver grafts with less than 10 minutes of warm ischemia can tolerance 12 hours of cold ischemia.More than 12 hours,postoperative liver transplantation complications ascend and,contradictorily,graft and recipient survival rate descend.

Objective To evaluate the effect of endoscopic retrograde cholangiopancreatography (ERCP) in treatment of biliary complications after liver transplantation. Methods Data of 39 patients who underwent ERCP between January 2005 and December 2007 because of biliary complications after liver trans-plantation were retrospectively evaluated. Endoscopic sphincterotomy, dilatation, nasal-biliary drainage (ENBD) and stent placement were performed in 25 patients with biliary strictures (14 at anastomosis site and 11 at non-anastomosis site). ENBD and stent placement were applied in 6 patients with biliary leakage, while endoscopic sphincterotomy, ENBD and stone extraction with baskets were performed in 16 patients with biliary stones. Procedures were repeated when necessary. Results ERCP was successfully performed at a rate of 95.9% (94/98) without any severe complications. Strictures at anastomosis site were resolved in all patients (100%, 14/14), while for strictures at non-anastomosis site, only 27.3% (3/11) were cured. Biliary leakage was resolved in 83.3% (5/6) patients. Complete bile duet clearance was achieved in 81.3% (13/16) of the patients with biliary stones. Conclusion ERCP proves to be safe and effective in the treatment of post liver transplantation biliary complications with low incidence of severe complications.

Objective:To develop a biomimetic nanoparticle probe of hematoporphyrin monomethyl ether (HMME) coated with breast cancer cell membrane, to observe its ability to target homologous breast cancer cells in vitro, and to investigate its effect of enhanced photoacoustic imaging and sonodynamic therapy (SDT) for breast cancer in vitro.Methods:The cell membrane of breast cancer 4T1 was extracted by chemical cleavage and repeated freezing and thawing. Then the HMME-coated polylactic acid-glycolic acid copolymer biomimetic nanoparticle was prepared by double emulsification and extrusion. The basic characteristics of nanoparticles were detected. The target ability of nanoparticles to homologous breast cancer cells and the enhancement of photoacoustic imaging were observed in vitro. Singlet oxygen sensor green (SOSG) was used to verify the reactive oxygen species (ROS) production of nanoparticles, and its SDT effect on breast cancer cells was evaluated by CCK8 cytotoxicity assay.Results:The size of the prepared CHP-NPs was uniform, the morphology was spherical "core-shell structure" , the particle size was (275.23±8.25)nm, and the surface potential was (-18.43±0.45)mV. It was observed that CHP-NPs could target homologous 4T1 cells under laser confocal microscopy. In vitro photoacoustic imaging experiments show that the photoacoustic signal of nanoparticles increases with the increase of its concentration. According to SOSG probe detection, CHP-NPs could produce ROS under ultrasonic irradiation.When CHP-NPs was incubated with 4T1 cells alone and no ultrasonic irradiation was used, the cell survival rate was not significantly affected. When the concentration was 0.6 mg/ml, the cell survival rate was still 95%. After ultrasonic irradiation, CCK8 experiment showed that the CHP-NPs had a significant SDT effect on breast cancer cells.Conclusions:The biomimetic nanomolecular probe of breast cancer cell membrane is successfully prepared. The probe has good ability to target homologous tumor, and can significantly enhance tumor photoacoustic imaging and SDT effect.

Objective To explore the clinical efficacy of the upper abdomen organ transplantation in the treatment of end stage liver disease and type 2 diabetes mellitus (T2DM).Methods The clinical data of 7 cases receiving liver-duodeno-pancreatic organ cluster transplantation in patients with end-stage liver disease and T2DM were retrospectively analyzed.The pancreas and the whole digestive tract of the recipients were reserved during operation,simple liver excision was executed,and abdominal multiple organs including pancreas,duodenum and part of jejunum were transplanted.The liver and kidney functions,blood glucose,C peptide,infection,rejection,vascular complications,biliary complications and other indicators were monitored postoperatively.Results No insulin was used in all the patients 1-7 days after operations,the blood glucose levels returned to almost normal, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin became normal after 1 week,and C peptide levels reached the normal range in 1 to 2 weeks.Among 7 patients,one died of graft-versus-host disease 1 month after operation,one got acute renal failure,one suffered from pulmonary infection, one had pancreatic leakage but recovered after unobstructed drainage,and no obvious complications were observed in the remaining patients.So far 6 live recipients were followed up for 1-39 months,the functions of the liver and pancreatic grafts were normal without hepatitis B and tumor recurrence, and the quality of life in the patients was satisfactory.Conclusion Liver-duodeno-pancreatic organ cluster transplantation is technically feasible and an effective method for the treatment of end-stage liver disease with T2DM.

ObjectiveTo investigate the effectiveness and tolerability of immunosuppressive regimen with daclizumab induction therapy. MethodsIn study group, 139 patients received immunosuppressive regimen with daclizumab induction therapy. In historical control group, 106 recipients received immunosuppressive regimen without daclizumab induction therapy. All patients were followed up at least for 1year. The acute rejection episodes, infectious and metabolic complications at one month and one year post-transplantation were compared between two groups.ResultsThe one-month incidence of acute rejection, new-onset diabetes mellitus, hypertension and infection was 7. 9 ％, 33. 8 ％, 21.6 ％ and 22. 3 ％, respectively in study group, as compared with 15. 1 ％, 72. 6 0％, 40. 6 ％ and 43. 4 ％, respectively in control group ( P ＜ 0. 05 ). The one-year incidence of acute rejection, new-onset diabetes mellitus, hypertension and hyperlipidemia was 10. 8 ％,5. 0 ％ ,4. 3 ％ and 7. 9 ％, respectively in study group, as compared with 19. 8 ％, 9. 4 ％, 8. 5 ％ and 14. 2 ％, respectively in control group (P＜0. 05). The one-year survival rate was comparable between two groups (P＞0. 05). ConclusionThe immunosuppressive regimen with daclizumab can enable early steroid withdrawal, significantly reduce acute rejection rate and various side effects mediated by longterm steroids use.

ObjectiveTo investigate a reasonable perioperative nutrition support therapy for combined ‘en bloc’ liver/pancreas transplants (CLPT).MethodThe clinical data of 10 patients,5 with gastrointestinal malignancy and liver metastasis and the other 5 with end-stage liver cirrhosis complicated with insulin-dependent type 2 diabetes mellitus ( T2DM),who had undergone CLPT in our center from May 2004 to September 2010 were retrospectively analyzed.All these patients received preioperative nutrition support,including normal food combined with nutrient solution before surgery,and total parenteral nutrition (TPN) to parenteral nutrition (PN) +enteral nutrition ( EN ) and to EN after surgery.The intestinal absorption,nutritional status,laboratory test results,and complications were recorded.ResultsAll recipients experienced a smooth recovery from the procedure,with normal or almost normal liver,pancreas,and duodenum graft functions.Three patients suffered from intestinal fistula,and all of them received TPN therapy; two patients died of multiple organ failure and one recovered from the complication.The remaining seven patients had smooth transition from TPN to EN.Of the 5 patients with malignancies,three died of multiple organ failure and 2 died of cancer recurrence.Of the 5 patients with liver cirrhosis and T2DM,four patients survived and 1 patient died of graft-verse-host disease.ConclusionRational perioperative nutrition support is important for the successful recovery after CLPT.

Objective To investigate the efficacy and safety of sirolimus in treating calcineurin inhibitor-related renal insufficiency after liver transplantation. Methods Eleven patients with calci-neurin inhibitor-related renal insufficiency after liver transplantation received sirolimus conversion.Simultaneously, the dose of tacrolimus was decreased or tacrolimus was withdrawn. Blood creatinine,sirolimus level, tacrolimus level, liver function, rejection episodes and drug side-effect were moni-tored. Results All the 11 patients survived today with 6 to 20 months of follow-up. All patients showed improvement of renal function after conversion to sirolimus. Blood creatinine level was reduced from (163.8±47.9)μmol/L to(108.1±26.6)μmol/l. (P<0.05). One patient's liver function had an acute rejection episode that was successfully treated with increase of dose of tacrolimus. The side-effects of sirolimus included hyperlipidemia (4 patients), anaemia (1) and mouth ulcers (2).Conclusion Siolimus can be effectively and safely used in liver transplant recipients suffering from ta-crolimus-related renal insufficiency.