Abstract:Abstract:Objective:To evaluate the safety of using so-called “marginal donors liver (MDL)” in liver transplantation, that included livers with steatosis, parasitic infestation,HCV(+) and HBsAg(+).
Methods :The clinical data of 85 cases of primary liver transplantation in our hospital were retrospectively analyzed. Among the donor livers, 44 were MDL including 35 livers with steatosis, 6 with parasitic infection, 2 with HBsAg(+), I with HCV(+). Of the 35 livers with steatosis, the grafts were divided into 3 subgroups based on the degree of fatty infiltration. Liver function parameters including AST, ALT, TB, PT on the day of surgery and 1,2,3,7 days after surgery were monitored among the 3 sub groups, as well as the incidence of delayed nonfunction (DNF), and 3 and 6 months patient survival rate. Six hepatic graft with parasite,1 with HCV(+) and 2 with HBsAg(+) were also closely monitored as above.
Results:There were significant difference in postoperative ALT、AST、TB、PT between hepatic steatosis≥40％ group and 20％~40% group, and between hepatic steatosis 20％~40% group and＜20% group. There was no significant difference among the groups in DNF occurrence rate and 3 month, 6 month patient survival rate in the 3 groups. Correlation analysis suggested that the degree of donor liver with steatosis was positively correlated with the damage of liver function after transplation. The use of HCV(+) and HBV(+) grafts did not lead to an increased mortality.
Conclusions:Grafts with ≥20％ steatosis are relatively safe donors. As the degree of severity of steatosis increases, the risk of post-transplantation liver failure is increased. Grafts with ≥40％ steatosis should be more carefully used or avoided as far as possible, expect in emergency because the higher the degree of graft steatosis the more serious is damage of postoperative liver function. The use of an HBsAg (+) and HCV (+) cadaveric liver graft and hepatic graft with parasite may be considered when no other donor is available.

Abstract:Abstract:Objective:To investigate the safety of different concentrations of FK506 for patients underging living donor liver transplantation (LDLT).
Methods :The clinical data of 46 LDLT recipients in recent 5 years were analyzed. The safety,immune complications and survival rate 1 month and 1 year after LDLT between the low concentration of FK506 (3-5ng/mL, 1year later 1-2 ng/mL) group and that of normal level (10-15 ng/mL, 1year later. 5-8 ng/mL) group were compared.
Results:At 1 month posttransplantation, the occurrence of acute rejection, hypertension,hyperglycemia and liver function was not significantly different between the two groups. Infection rate and serum creatinine level was significantly lower in low concentration groups than normal concentration groups at 1 month and 1 year. The graft survival rate was not different between the two groups at 1 month and 1 year.
Conclusions:A low concentration of FK506 in perioperative and long-term period is safe. The infection rate and the level of serum creatinine after transplantation are positively related to blood concentration of FK506.

Abstract:Abstract:Objective:To summarize the clinical experience of standard piggyback liver transplantation (SPBLT) and modified piggy-back liver transplantation (MPBLT) in the treatment of Wilson′s disease (WD).
Methods :The clinical records of 29 cases of WD who underwent piggyback liver transplantation over the recent 12 years were analyzed retrospestively. Among them, there were 22 male and 7 female patients, with 6cases aged 8-14y and 23 cases aged 24-37y, who underwent liver transplantation because of chronic advanced liver disease (18cases), fulminant hepatic failure (2 cases), or with normal liver function(9 case), and among them, 24 cases showed neurological dysfunction. The modes of operation included SPBLT (13 cases) and MPBLT (16 cases).
Results:The patients were followed up for 1 month-12 years with the median follow-up time of 47 months. Four patients died in the perioperative period,of which 2 died of hepatic failure due to obstruction of hepatic venous return after SPBLT,and 2 died of hepatic artery thrombosis and bile leakage combined with severe infection.The recipient survival rate at 1 year and 3 years was 86% and 79% respectively.One patient has survived for 12 years.All the levels of serum copper and copper-protein recovered to normal in four weeks post-operatively. Neurological symptoms improved in varying degrees after operation.
Conclusions:WD patients who undergo PBLT can expect a satisfactory clinical outcome and a good quality of life, and MPBLT can effectively reduce the complications of operation and improve the operation survival rate.

Abstract:Abstract:Objective:To investigate the general rule of perioperative hemodynamic changes in liver transplantation patients.
Methods :We retrospectively studied the data of 31 patients who were treated with liver transplantation from July 2007 to December 2007. K value, EHBF, CO, BV and CBV were detected by DDG analyzer on preoperative day and on 1, 3, 7 and 14 postoperative day.
Results:K value, EHBF, CO, BV and CBV detected before operation was （0.09±0.05）/min, （0.43±0.22）L/min, （7.16±2.49）L/min, （5.87±1.85）L, （3.94±1.59）L respectively. Postoperatively, K value and EHBF were significantly higher than that of preoperative day; CO and BV of the 1 postoperative day were significantly higher than that of preoperative day and the 3 and 14 postoperative day; while CBV of the 1 postoperative day was higher than that of other days(P＜0.05).
Conclusions:Before liver transplatation operation the patient has low blood volume and high circulation. In the early postoperative period has high blood volume, high circulatory state, increase of effective hepatic blood flow and rapid recovery of hepatic reserve function. At 3d postoperatively, the high circulatory state markedly decreases, and general circulation gradually enters a stable condition at about 2 weeks after operation.

Abstract:Abstract:Objective:To investigate the safety and therapeutic effect of different warm ischemia time (WIT) of non-heart beating (NHBD) liver graft in clinical liver transplantation.
Methods :A retrospective study on 142 NHBD cases which were under different WIT and cold ischemia time (CIT) within 12h was carried out. According to different WIT, the grafts were divided into 3 groups: group Ⅰ, less than 5min (43 cases); group Ⅱ,5-10min (77cases); group Ⅲ, 10-15min (22 cases). The difference in 3 groups in the aspects of the peak value of alanine aminotransferase (ALT), primary liver graft nonfunction (PNF), acute rejection(AR), biliary complications, vascular complications, infection, graft survival rate and recipient survival rate after liver transplantation were compared.
Results:No PNF occurred in 3 groups post-transplantationly. In group Ⅱ only the peak value of AST and infection rate were significantly higher than group I; but there were significant differences between group Ⅲ and group Ⅰin the aspects of peak value of AST, biliary complications, vascular complication, infection rate. The graft survival rate and recipient survival rate in group Ⅲ were significantly shorter than group I and group Ⅱ in follow up time was 8-23 months.
Conclusions:For NHBD with CIT less than 12 h, the maximal safe duration of WIT is 10 minutes. If WIT is over 10 minutes, there will be a significant higher rate of correlative complications, lower graft survival rate and recipient survival rate after liver transplantation.

Abstract:Abstract:Objective:To investigate the methods of hepatic artery(HA) preparation and reconstruction in donors with HA anatomic variations in orthotopic liver transplantation.
Methods :A retrospective analysis was applied to find the methods and skills of donor liver and hepatic artery(HA) reconstruction in the 91 cases of orthotopic liver transplantation.
Results:All allografts prepared were used for liver transplantation. Twenty of 91（21.9％）allografts had hepatic artery anatomical variations, and reconstructive anastomosis was performed in 12 cases. Splenic artery（7/12）and gastro-duodenal artery（5/12）were used for anastomosis to the variant right hepatic artery. No complications resulted from donor liver preparation postoperatively.
Conclusions:Correct donor liver preparation and hepatic artery(HA) reconstruction in donors can decrease the incidence of hepatic artery and biliary complications.

Abstract:Abstract:Objective:To study the influence of combined anti-CD-40L and low dose CsA immunotherapy on the induction of immune tolerance in orthotopic liver transplantation rats.
Methods :Recipients were divided into the following groups. Group A,SD→SD; Group B,SD→Wistar without any treatment; Group C,SD→Wistar with CsA therapy from d1 to d5; Group D,SD→Wistar with CsA from d1 to d5 and anti-CD40L mAb in d0 and d2, and Group E,SD→Wistar with CsA from d1 to d5 and anti-CD40L mAb in d0 and d2 plus autotransfusion before operation. The recipient survival time, histologic changes of allograft and the level of cytokines in peripheral blood were observed.
Results:The survival period of recipients in Groups A, D and E was significantly longer than that in Groups B and C. The acute rejection response in Groups D and E decreased prominently. The serum level of IL-2 and IFN-γ in Group B were significantly higher than those in other groups, while the serum level of IL-4 and IL-10 in Groups D and E was elevated much more than those in Group B(P＜0.05).
Conclusions:The combined anti-CD-40L and low dose CsA immunotherapy with or with autotransfusion can prolong graft survival time, suppress acute rejection and induce high expression of Th2 cytokines, and is beneficial to long-term survival of recipients and liver graft.

Abstract:Abstract:Objective:To explore the impact and significance of hypoxia preconditioning on the expression of Bcl-2 protein after orthotopic liver autotransplantation in rats.
Methods :A modified orthopotic liver autotransplantation model was used to study the ischemia reperfusion injury in liver transplantation. Sprague-Dawley rats were randomly divided into the following three groups: normal control (NC) group, autotransplantation (AT) group, and hypoxia preconditioning (HP) group, there were twenty four rats in each group. HP Group was given an 8% oxygen -mixed gas for 90 minutes before the operation. At 1, 6, and 24 hours after the operation, the rats were killed and the following tests were conducted: (1)Liver tissue was sampled to observe the expression of Bcl-2 protein; (2)blood was drawn to conduct a chemical examination; (3) morphology was observed by light microscopy and transmission electron microscopy; (4)chondrosomes were quantitatively analysed by a MiVnt image analysis system.
Results:The serum levels of ALT and AST in HP group were significantly lower than that of AT group (P＜0.05) at 1, 6 and 24 hours after the orthopotic liver autotransplantation. The expression of Bcl-2 protein was increased significantly (P＜0.05) in HP group at each measurement point. The hepatic tissues in HP group were less swollen than AT group under light microscopy. Hepatic cells in AT group showed typical apoptosis signs under transmission electron microscopy (TEM), but no apoptosis was found in HP group. The appearance of hepatocytes in HP group was much better than AT group, and the areas, perimeters and diameters of chondrosomes in group HP under TEM were much smaller than that in AT group (P＜0.05).
Conclusions:HP has marked inhibition to apoptosis by up-regulating the expression of Bcl-2 protein and protection to chondrosomes after an orthopotic liver autotransplantation.

Abstract:Abstract:Objective:To explore the feasibility of constructing PTLD model.
Methods :The peripheral blood lymphocytes of 40 EBV(+) volunteers were injected intraperitoneally into 3 SCID mice respectively. IgG determination was utilized to confirm the reestablishment of human immunocell in SCID mice. The tumor tissue of the dying or dead mice were retrieved,and then the establishment of LPD model was determined by pathological section and immunohistochemistry.
Results:The IgG level in the blood serum of SCID mice was (750.0±13.2)μg/mL and (1 050.0±14.6)μg/mL respectively at 8 weeks and 12 weeks post transplantation. Pathological section of retrieved tumor tissue indicated that the tissue was large-cell lymphoma with high mitotic rates.The tumors were CD20 positive and LMP-1 positive. Among the 40 volunteers, human peripheral blood lymphocyte from 10 volunteers (25%) produced LPD rapidly(median time to LPD, 8 weeks)and with high penetrance (median, 100%); whereas 20% (8 of 40 volunteers) developed LPD tumor later (median 12 weeks),and only in partial mice (median penetrance, 55%); HuPBLs from the remainder of the donors(22 of 40,55%) produced no LPD after 16 weeks.
Conclusions:PTLD model can be successfully established by intraperitoneal injection of EBV(+)huPBL in SCID mice.

Abstract:Abstract:Objective:To comparatively investigate the protective effect of HGF and SAM on liver regeneration and liver function in cirrhotic rats after hepatectomy.
Methods :Liver cirrhosis of the rat was successfully induced by injection of CCL4.Then 30% partial hepatectomy was performed in all rats.Cirrhotic rats were divided into 4 groups, namely, cirrhotic group(normal saline 5 mL/d IM, n=16), HGF group(HGF 0.65 mg/(100 g·d) peritoneal injection, n=16), SAM group（SAM 20 mg/(kg·d) IM, n=16）and HGF+SAM group(HGF 0.65 mg/(100g·d), SAM 20 mg/(kg·d), n=16).Animals were sacrificed on the 15th post-operative day to take samples for detection of liver function(AST, ALT, ALB and TB).Liver tissues were also observed under light microscope and electron microscope.
Results:In HGF group, SAM group and HGF+SAM group, valus of AST, ALT and TB were decreased significantly as with cirrhotic group(P＜0.05), and concentrations of ALB were increased significantly(P＜0.05).
Conclusions:The study suggests that the liver function and regeneration of cirrhotic rats have impaired after partial hepatectomy. However HGF and SAM have the efficacy of enhancing liver regeneration and improving liver function.

Abstract:Abstract:Objective:To study the biological effects of silencing of Rel-A gene expression by RNA interference on the dendritic cells.
Methods :Mouse dendritic cells derived from bone marrow were amplificated through cytokine inducing system and purified with positive immunomicrobeads by flow cytometer.The mRNA and protein expression, aiming directly at Rel-A, were analyzed by semi-quantified RT-PCR and Western blot. A pair of the highest effective siRNA were synthesized selectively and packed by lentivirus transfer vector, then transfected to DC(RNA interference group), while control group was not transfected by RNAi; 12 hours later, lipipolysaccharide (LPS) at a concentration of 1mg/L was added to two groups and continued to culture. The cell microstructure was then observed under transmission electron microscopy. The cell surfacemarkers, such as MHC Ⅱ, CD86 and CD80, were detected by flow cytometer. The functional properties of DCs were detected by mixed lymphocyte reaction (MLR). The concentration of TNF-α, IFN-γ and IL-12 in the supernatant were detected by ELISA.
Results:mRNA and protein expression were reduced 90% in sequence 2 siRNA, the difference was significant statistically, in comparison with control group. Many phagocytic vacuoles were observed in the cytoplasm under transmission electron microscopy. The expression of CD80, CD86 and MHC Ⅱ in the cytomembrane of DCs,the proliferation of allogeneic T cells and the concentration of TNF-α, IFN-γ and IL-12 decreased more significantly in RNA interference group than in control group.
Conclusions:RNA interference can reduce Rel-A expression in murine myeloid dendritic cells and inhibit the maturation of DCs. The dendritic cells interfered with silencing of Rel-A gene are characterized as the biological features and immunological function of immature DC. This kind of RNAi Rel-A DCs may be used to induce immunotolerance as novel tolerogenic DCs.

Abstract:Abstract:Objective: To investigate the change of the invasive and metastasis ability of HepG2 cells by silencing BC047440 genes.
Methods :The experiment were divided in 3 groups: in terfering group, HepG2/HK group and HepG2 group.Small interfering RNAs for BC047440 gene were initially designed and silenced to BC047440 genes, then the cell adhesion was measured with MTT assay, the cell migration was surveyed by scratch method, and the cell invasion in vitro was determined with Boyden chamber.
Results:There was no significant difference of the adhesion rate in the interfering group compared with HepG2/HK group and HepG2 group at 20 min and 40 min after seeding, while there was significant difference at 60min (P＜0.05). The scratch wound had almost been filled with HepG2/HK group and HepG2 group at 48h after scratching, while it was still apparent in the interfering group. The number of cells that penetrated polycarbonate coated with matrigel when incubation for 16h in interfering group was much less than HepG2/HK group and HepG2 group (P＜0.05).
Conclusions:The silencing BC047440 genes could significantly inhibit the ability of adhesion to extracellular matrix, the ability of migration to scratch wound, the ability of invasion of HepG2 cells in vitro, and these results suggest that BC047440 could regulate the invasive and metastasis ability of HepG2 cells.

Abstract:Abstract:Objective:To construct the double-H1promoters siRNA expression cassettes (SECs) targeted to human telomere retrotranscriptase (hTERT), and investigate the interfering effect of its siRNA on hTERT gene expression in HepG2 cells.
Methods :SECs were constructed by fusing PCR, based on two different human telomerase hTERT gene fragments.When SEGs transferred into HepG2 cells respectively, the SECs were transcripted to the siRNA.The interfering effect of SECs on the telomerase activity in the cells was assessed by telomeric repeat amplification protocol (TRAP) and PCR-EIA.
Results:SEGs were successful constracted, and the telomerase activity was significantly inhibited when the HepG2 cells were tranfected with SECs.
Conclusions:The siRNA SECs display a definite RNA interference effect on the expression of telomerase. This method of SECs preparation can be applied for RNAi research in tumor inhibition.

Abstract:Abstract:Objective:To explore the relationship and interaction between extracellular matrix (ECM) components and oval cells (OC) during the oval cells-mediated liver regeneration.
Methods :A dynamic study on the localization of and the relationship of OC with ECM components (laminin and fibronectin) during the proliferation and differentiation of OC in 2-AAF/PH rats model was conducted and analysed by using immunohistochemistry and double immunofluorescent method.
Results:By day 2 after partial hepatectomy, OC began to proliferate around the portal area.Most of laminin were present along the hepatic sinusoids in the periportal area. Fibronectin markedly increased in the whole hepatic lobule.From day 4 to 9,OC spread further into hepatic parenchyma,closely associated with fibronectin and laminin.There was a decrease in number of the pericentral fibronectin and increase of fibronectin in the periportal areas. From day 12 to 15, as the OC differentiated into small hepatocellular nodus, most of laminin and fibronectin were located around the hepatocellular nodus, and few of them were present into the nodus.After day 18, the normal liver lobule structures began to recover.
Conclusions:There is a close relationship between the ECM components and OC during the liver restitutive repair. Local hepatic microenvironment may participate in the oval cell-mediated liver regeneration through the cell-matrix interactions.

Abstract:Abstract:Objective: To find an ideal liver cirrhosis portal hypertension model in big animals suitable for surgical study.
Methods :Twenty canines were treated by subcutaneous injection of 60% CCl4 colza oil emulsion into the back, at a dose of 1.0-1.3 mL/kg every 10 days for a total of 6-8 times. At the same time, it was combined with the control of food intake. All canines were fed with rice mixed with 10% hog fat. The amount of rice was 15 g/kg per day from the first day to the forth day, but the amount of food was not controlled from the fifth day to the tenth day. During the process, the canines′ general condition was observed and the changes of hepatic functions such as ALT, TP, ALB, G and A/G were measured. The changes of liver morphology, the diameter and blood flow of portal vein were monitored with color Doppler. Every two weeks, a piece of hepatic tissue to observe the pathological change of liver was excised operatively.
Results:During the process, the body weight of the canines decreased continually; ALT increased during early and middle stages and decreased during advanced stage; TP and ALB always decreased, and A/G continuously decreased, but G did not change significamtly. Eight weeks after injection, liver became progressively smaller. its density was not well-distributed, and liver particles gradually became coarse, and liver capsule became rough. The diameter of portal veins became larger and the velocity of portal vein became slower. Liver cirrhosis with psudolobules was found on light microscopy from the tenth week to the twelfth week. Three canines died, with mortality of 15.0%.
Conclusions:It took 10-12 weeks to establish this liver cirrhosis portal hypertension models in canines by subcutaneous injection of CCl4. This method is convenient, has high success rate, and is suitable for use in surgieal research.

Abstract:Abstract:Objective:To study the diagnostic methods and feasibility of surgical treatment of giant hepatic cavernous hemangioma.
Methods :The clinical documents of 31 patients with giant hepatic cavernous hemangioma were retrospectively analyzed. All of the patients were examined with image technology before surgical operation. Among them, 16 patients were examined with B-US and MRI, 11 patients with B-US and CT, and 4 patients with B-US, CT and MRI; and 29 patients were confirmed as hepatic cavernous hemangioma (93.6%), but 2 patients were misdiagnosed as hepatic cancer (6.4%). All patients underwent radical surgical operation.
Results:There was no opeative death, and all these patients had successful resection of giant hepatic hemangioma. The average amount of hemorrhage and blood recovered from patients during operation was 800 mL (400-2 000 mL) and 600 mL (250-1 500 mL) respectively. Two patients suffered from biliary tract injury during operation; and after operation. 18 patients experienced pleural effussion, 6 patients had subphrenic fluid collection, 4 patients suffered from pneumonia.All patients were cured and discharged.
Conclusions:The most suitable preoperative examination for patients with giant hepatic hemangioma is B-US together with MRI. A detailed analysis of image data of hemangioma, a well designed operative plan and accurate management during operation are key points to successful operation.

Abstract:Abstract:Objective:To review the experience in perioperalive management of 324 consecutive cases of pancreatico-duodenectomy (PD).
Methods :The clinical data of 324 cases of PD were analyzed retrospectively. All underwent PD successfully, 275 cases received the standard PD, while, 49 cases had pylorus preserving PD.Digestive tract reconstruction was done by Child method. Pancreatointestinal anastomosis was performed by pancreas stump intussussception in 303 cases, and end-to-side anastomosis in 21 cases.
Results:The 30 day mortality rate was 0.3% (1/324). Complicalion rate was 8.0%, included one case of liver dysfunction, 7 cases with pancreatic fistula and intraabdomind bleeding, 1 case of stenosis of pancreaticoenteral anastomosis, 3 cases with pancreatic dysfunction, 4 cases with gastroenteral anastomosis ulcer, 6 cases with delayed gastric emptying, 1 with reflux of bile and 3 with lymphatic fistula.
Conclusions:Perioperative managemen for pancreaticoduodenectomy is crucial for patient prognosis. Carful perioperative management can decrease postoperative complications and ensure smooth recover of these patients.

Abstract:Abstract:Objective:To investigate early diagnosis and treatment of tumor of duodenal papilla.
Methods :Their clinical data of 64 cases of tumor of duodenal papilla confirmed by operation and pathology were analyzed retrospectively. They included 60 cases of carcinoma of duodenal papilla and 4 cases of adenoma.
Results:The main clinical symptoms were jaundice(42 cases), digestive tract symptoms (35 cases),recurrent cholangitis(7 cases) and hemorrhage of upper digestive tract(1 case). The diagnosis rate accuracy of ERCP,Fiberoptic duodenoendoscopy，MRCP,BUS and CT were 100%,97.3%,82.4%,82.8% and 76.1% respectively.Fifly-five cases underwent pancreatoduodenectomy,5 cases received local resection, and the remaining 4 cases were treated by palliative surgery. The surgical complication rate was 16.1% and the surgical mortality rate was 3.6%. The 1-year,3-year and 5-year survival rate of pancreatoduodenectomy were 67.4%,40.6% and 36.3%.
Conclusions:Jaundice and abdommal pain are the main symptoms of tumor of duodenal papilla.Fiberoptic duodenoendoscopy and ERCP are most effective methods for diagnosis of tumor of duodenal papilla.It is essential to early select radical resection operation so as to improve the results of surgical treatment.

Abstract:目的:探讨经腹食管下段胃底切除术治疗门静脉高压症的疗效。
方法:对2004年1月—2007年6月应用经腹食管下端、近端胃部分切除治疗门静脉高压症的19例患者(切除组，A组)的临床资料进行回顾性分析，并与同期行传统贲门周围血管离断术的36例患者（断流组，B组）进行比较。
结果:A组复查上消化道钡餐均示食管静脉曲张消失，术后2～3年未发现有消化道再出血者； 而B组食管静脉曲张复发率和再出血率分别是31.3%和14.3%，两组比较P值分别为P＜0.05, P＜0.01。两种术式肝功能、血常规改善比较，差异无统计学意义。A组术后2～3年无吻合口瘘和吻合口狭窄发生。
结论:经腹食管下段胃底切除术是治疗门静脉高压症的一种有效术式。