Objective　 To explore the feasibility of recombinant adeno-associated virus (rAAV) as a vector for the gene therapy of liver cancer. Methods　 The rAAV/enhance green fluorescein protein (EGFP) recombinant was prepared by the routine method of two plasmids cotransfection. Results　 The experiment showed that one 10cm plate could produce 107-108 infection unit recombinant by the method of two plasmids cotransfection, and the transduction of HepG2 cell was increased with the increase of infection dosage of rAAV. About 100 multiplicity of infection (MOI) AAV vector could make all the tumor cell light. Conclusion　 Liver cancer cell can be efficiently transduced by rAAV, and AAV vector may be a valuable vector for the gene therapy of liver cancer.

Objective To observe the efficacy and adverse reaction of the test drug hepatitis B immunoglobin on the post market.Method Employed by the methods of multiple center's clinical trials and using the recommended dosage of hepatitis B immunoglobulin for intravenous administration,the clinical efficacy of either prevention or treatment for hepatitis B recurrence and drug related adverse reactions were observed.This consisted of 22.1 months,13 hospitals,and 525 patients with hepatitis B related liver transplantation.Result The results showed a contrasting probability of adverse reactions for different doses among the observation period.Within 6 months postoperatively with a greater or equal to recommended drug dose,the infection rate was less than 4％.In contrast,the infection rate was greater than 12％ in the group with less than the recommended drug dose.Conclusion There was an obvious dose effect relationship,and the drug safety and recommended treatment rationality were verified.

Objective To summarize the experience in hepatic artery reconstruction in adult-to-adult living donor liver transplantation(ALDLT).Methods Fifty patients underwent ALDLT in our hospital from January 2002 to July 2006.All the hepatic a~ery reconstructions were done under surgical microscope.ResultsTwo patients(4%)presented with hepatic artery thrombosis.All the patients were followed up for 2 to 52 months (median,9 months),and no hepatic artery stenosis nor hepatic artery pseudoaneurysm occurred.The 1-year survival rate was 92%(46/50).Conclusions Systematic evaluation of hepatic artery reconstruction and use of microsurgical technique are key to the reduction of complications of hepatic artery reconstruction in ALDLT.

Objective To determine the value of helical computed tomography (HCT) and color doppler flowing imaging (CDPI) in evaluating the resectability of pancreatic cancer. Methods The clinical data of 114 patients with pancreatic cancer who had been admitted to the Affiliated Hospital of North Sichuan Medical College from January 1995 to December 2002 were retrospectively analyzed. The values of HCT and CDPI in assessing the resectability of pancreatic cancer were determined according to the results of operation and pathological examina-tion. All the data were analyzed by chi-square test and Fisher exact probability. Results Of all patients, 109 were examined by HCT, 97 by CDPI and 96 by HCT+CDPI. For patients examined by HCT, the resection rates of pancreatic head cancer, pancreatic body and tail cancer and total pancreatic cancer were 45.3% (39/86), 26.3% (5/19) and 0 (0/9), respectively. The resection rate of pancreatic head cancer was higher than that of pancreatic body and tail cancer (χ2=8.965, P<0.05). With the increase of tumor size, the invasion rate and metastasis rate were increased and the resection rate was decreased (z=6.15, 5.35, 7.18, P<0.01). The sensitivity rate and specificity rate were 77.8% and 82.2% of HCT, 73.3% and 80.6% of CDPI, 90.6% and 92.4% of HCT+CDPI in assessing the resectability of pancreatic cancer. The values of Kappa identity test of HCT, CDPI and HCT+CDPI were 0.58, 0.52 and 0.82, respectively. Conclusions Combined application of HCT and CDPI can further improve the accuracy in assessing the resectability of pancreatic cancer.

Objective To construct the expressing vector of shRNA/mrp1 and study its expression in vitro. Methods 64bp oligonucleotides of pSUPER and the targeted senquence of siRNA/mrp1 were synthesized and annealed to form duplex strand, then were cloned into pSUPER to construct pSUPER-shRNA/mrp1 vector. Competenced Ecoli was transfected by vector of pSUPER-shRNA/ mrp1 to screen the positive clones for sequencing and extracting plasmid. The plasmids extracted were used to transfected HepG2/mrp1 cells with a control groups by negative vectors. The expression of mrp1 mRNA and MRP1 was measured by real-time PCR and resistance of HepG2/mrp1 by flowcytometry. Results pSUPER-shRNA/mrp1 was established successfully and was sequenced to test its accuracy. Expression of mrp1 mRNA in HepG2/mrp1-si was lower than that in HepG2/mrp1 (1-fold vs 179.76-fold, P<0.001). Compared to HepG2/mrp1, the expression of MRP1 in HepG2/mrp1-si was lower (11.2% vs 97.6%, P<0. 05). The sensitivity of HepG2/mrp1-si to adiramycin was higher than that of HepG2/mrp1(45.0-fold vs 1.2-fold, P<0.01). Meanwhile, the accumulation of DNR in HepG2/mrp1-si increased significantly as compared with the control (78.58 % vs 38.44%,P<0.05).Conclusion Vector of pSUPER-shRNA/mrp1 can be constructed by the technique of enzymatic incision. The multidrug resistance of HepG2/mrp1 can be reversed by RNA interference.

Objective To discuss the influence of anatomical variations of the cystic duct on preoperative diagnosis and operational scheme for cholecystectomy. Methods A 47-year-old woman was admitted to our hospital with diagnosis of cholecystolithiasis. Ultrasonography suggested minimal intra- and extrahepatic ductal dilatation. Laboratory tests showed that serum levels of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase were 189 IU/L, 366 IU/L and 144 IU/L, respectively. In order to make a certain diagnosis, the patient received both magnetic resonance cholangiopancreatography (MRCP) and endoscopic retrograde cholangiopancreatography (ERCP). Results MRCP showed the bile duct slightly dilated with a shuttle shape figure and a lower signal with a strip form in it. MRCP could not confirm the quality of this signal and was doubtful of choledochus diaphragma. Subsequently, ERCP was applied to demonstrate that the cystic duct was collateral with the common hepatic duct when arriving into its left side and converged into the bile duct with a lower position, which was the reason for why MRCP misjudged the formation of choledochus diaphragma in the bile duct. Finally, the patient underwent open cholecystectomy. Conclusion There are some kinds of variations in the cystic duct including course, appearance and location of confluence. Combing MRCP with ERCP can significantly elevate the diagnostic accuracy of the cystic duct before operation, especially in those patients with doubtful diagnosis upon admission. To avoid biliary injury as much as possible, open cholecystectomy is superior to the laparoscopic cholecystectomy (LC)with regard to the patients suffering from cholecystolithiasis complicated with variation of the cystic duct.

Objective To evaluate the suitability of reported standard liver volume formulae for Chinese adults based on the practice of 216 cases of living donor liver transplantation in our transplantation center. Methods The graft volume was preoperatively estimated in 179 adult-to-adult right liver living donors by two methods: first, the radiological right liver volume by computed tomography (CT) and second,calculated graft volume obtained by reported standard liver volume formula and the percentage of the right liver volume ( given by CT). Both results were compared to the actual graft volume measured during surgery.Results The mean percentage of right liver volume was 55.4% (SD 5.41%). The results of Urata、Heinemann、Vauthey、 Lee、 Yoshizumi formula were significantly larger than the actual right liver volume (P ＜0. 01 ). The result of Sheung-tat Fan was less than the actual right liver volume, there was statistical ESLV =334. 024 + 11. 508 × BW, is most suitable to estimate adult Chinese donor's right liver volume.

Objective To investigate the liver function injury and the rate of complications in living liver transplantation donors in different graft type transplantation.Methods Postoperative data of 154 living liver donors satisfying our inclusion criteria were prospectively collected and registered from Jan 2002 to May 2009 in our hospital.We divided the donors into two groups (right-lobe graft, R group and left-lobe graft, L group), and made comparison on the liver function and complications.Results Remnant liver weight in R group were smaller than those in L group (t = 11.418, P ＜ 0.05).the ratio of remnant liver weight to standard liver weight in R group were smaller than those in L group (t = - 5.040, P ＜ 0.05 ) .Peaks of ALT, AST and INR in both groups appeared on the first day after operation, while the peak of TB in R group appeared on the third day after operation.All the index values returned to a normal baseline after reaching its peak.Mean values of TB in R group were higher than those in L group on the 1st, 3rd, 7th day after operation (seperately t1 = 5.285, t3 = 3.747, t7 = 2.729, all P ＜ 0.05).Mean values of INR in R group were higher than those in L group on the 1st, 3rd, 7th day after operation (seperately t1 = 5.260, t3 = 5.035, t7 = 2.267, all P ＜ 0.05).The level of TB in both groups returned to normal range on the 7th postoperative day, while the level of ALT and AST remained twice the upper limits of the normal.There were no deaths; Complications occurred in 53 of 154 donors (34.42% ) , 52/141 (36.88% ) in R group and 1/13 (7.69% ) in L group (x2 = 3.292, P ＞ 0.05).Conclusions Ramnant liver function of R group during early postoperative period was poorer than that of the L group.Donors were safe, though suffering from comparatively high complication rate.

Objective To observe changes of hepatic hemodynamics and hepatic functional reserve after splenectomy and periesophagogastric devascularization. Methods From July 2006 to August 2007, thirty patients with portal hypertension caused by hepatitis B cirrhosis underwent splenectomy and periesophagogastric devascuiarization. The PVPG (portal venous pressure gradient) was measured by inductor continually during operation. The HAF (hepatic artery flow), PVF (portal venours flow) and hepatic arterial RI(resistant index) were measured with Doppler sonography. The EHBF(effective hepatic blood flow) and ICGR15 (indocyaninegreen retention rate at 15 minutes) were obtained respectively by indocyaninegreen clearance test before and after the operation. Results PVPG after laparotomy (19±4) mm Hg, ligating the splenic artery(14±4) mm Hg, splenectomy(14±3)mm Hg and periesophagogastric devascularization (12±4) mm Hg showed a tendency to decrease progressively. The PVF decreased [from (42±14) ml/s to (16±8) ml/s] and HAF increased in compensation after operation. The EHBF increased [from (0.48±0.10) L/min to (0.56±0.10) L/min], and the ICGR15 decreased (from 23%±8% to 18%±4%) postoperatively. Conclusion After splenectomy and periesophagogastric devascularization, the hepatic functional reserve improves at least in a short term notwithstanding the decrease of PVPG and PVF.

Objective To investigate the levels of oxidative stress in liver tissues of hepatocelluar carcinoma(HCC)patients after transcatheter arterial chemotherapy(TAC).Methods Immunohistochemistry streptavidin biotinylated peroxidase(S-P)method was used to detect the cellular levels of 8-hydroxy-2'-deoxyguanosine(8-OHdG),p53 and p21~(waf1/cip1).Eighty-nine HCC patients were divided into TAC group(39 cases)and Non-TAC group(50 cases).15 Non-HCC liver tissues served as controls.Result 8-OHdG level was higher in Non-TAC group than that in TAC group in tumor tissues (F=9.516,P＜0.05),with that being the lowest in control group(F=9.516,P＜0.01);8-OHdG levels in cancer tissues were significantly higher than that in tumor surrounding tissues in both TAC group (t=7.101,P＜0.001)and Non-TAC group(t=8.020,P＜0.001),there was no significant difference of 8-OHdG levels between para-tumor tissues and controls.The levels of 8-OHdG between tumor and its surrounding tissues in TAC group(r=0.651,P＜0.001)and non-TAC group(r=0.493,P＜0.01)was in positive correlation.The difference of p53 levels in cancer tissues in TAC group and Non-TAC group were not statistically significant and p53 was not detected in para-tumor tissues.The difference of p21~(waf1/cip1) levels among TAC group,Non-TAC group and controls was statistically significant,the levels of p21~(waf1/cip1) in normal group was the highest(F=13.459,P＜0.001),followed by that in TAC and Non-TAC group in cancer tissues(TAC vs.Non-TAC group,P＜0.01);p21~(waf1/cip1) expression in normal controls was significantly higher than that in both TAC and Non-TAC group in para-tumor tissues(F=16.613,P＜0.001).The correlation of p21 ~(waf1/cip1) levels between tumor and its surrounding tissues was significant in non-TAC group(r=0.872,P＜0.001).Conclusions Oxidative stress levels in HCC tumor tissues were higher than in para-tumor tissues and non-HCC liver tissues.Cancer cells probably survive chemotherapy by fortifying oxidative stress repair mechanism.