Objective To study the effect of HCV C gene transfection on expression of hTERT mRNA in human biliary carcinoma cell lines (QBC939) and to elucidate the significance of activation of hTERT mRNA by HCV C gene on the carcinogenesis of bile duct cells. MethodsThe recombinant plasmid(pcDNA3-HCVC) and the vector-alone were co-transfected with enhanced green fluorescent protein( EGFP )into QBC939 and human normal bile duct epithelial cells(HBEC) using liposome. The reverse transcription PCR(RT-PCR) and immunocytochemical stain were used to show the expression of hTERT mRNA and protein. Results The transfection efficiency of pcDNAHCV-C,which was determined by the expression of EGFP,is about 16% and 30% in QBC939 and HBEC respectively. There was no expression of hTERT mRNA assayed in HBECs when transfected blank vector,but a dramatic increase was observed for hTERT mRNA expression in HBEC when transfected with HCV C expressing vector. The expression of hTERT mRNA and protein assayed in QBC939 significantly increased when transfected with HCV C expression vector than that transfected with blank vector. Conclusion HCV C gene transfection up-regulates the transcriptional expression of hTERT mRNA in biliary carcinoma cells,it is suggested that HCV C protein contributes to virus carcinogenesis.

Nuclear factor-kappa B (NF-κB) is a key regulator in epithelial-mesenchymal transition (EMT) of cancer cells.EMT of cancer,one of the reasons of drug resistance,enhances the infiltration and distant metastases ability of cancer cells.Recent researches show that Snail,Slug,Twist and Zeb play important roles in regulating EMT of cancer cells.Drugs and targeted therapies that inhibit NF-κB activities can reverse the EMT of cancer cells.NF-κB may become an effective therapeutic target in cancers in the future.

Objective To evaluate the efficacy of splenic autotransplantation plus lower esophagus transaction for the treatment of portal hypertension(PTH).Methods Thirty patients were divided into study group(15 cases)and control group(15 cases).Patients in study group Underwent splenic autotransplantation after splenectomy and cardia-esophageal devascularization plus lower esophagus transaction,and those in control group had all except splenic autotransplantation.The cross section area,blood velocity,blood flow of MPV(main portal vein)and changes of cardia-esophageal varices were evaluated by 3D DCE MRA at 1 week before operation and 6 months after,and blood flow and collateral circulation of transplanted spleen in the retroperitoneal space were assessed.Results In both groups,the cross section areas(cm2),mean blood velocity(cm/s)and mean blood flow(ml/s)of MPV decreased postoperatively(P＜0.05).The postoperative cross section areas(cm2)and mean blood velocity(cm/s) of MPV in study group were smaller than that in control group(P＜0.05).Esophageal and fundal variceal veins disappeared or improved equally in both groups.There was no difference in the postoperative and preoperative liver function between the two groups(P＞0.05).In study group,the planted spleen grew well in the retroperitoneal space,and with a formation of extensive collateral circulation.The postoperative serum hyaluronic acid decreased in this group(t=2.929,P＜0.05).Conclusion Splenic autotransplantation after splenectomy plus lower esophagus transection was effective for the treatment of PHT without adverse impact on liver function.

Objective To analyze the major causes of reoperation for abdominal injuries and explore the effective preventive measures so as to enhance operation quality for abdominal injuries. Methods The data of 33 patients undergoing relaparotomy for abdominal injuries during the last 10 years was analyzed retrospectively. Results There were 25 closed injuries and 8 stab penetrating injuries in this group. The causes of reoperation were mainly missed injuries in initial laprotomy 10 cases,inappropriate surgical procedures 8 cases, postoperative complications 13 cases, and negative relaparotomy 2 cases. In these 33 patients, 40 times of reoperation were performed, and among them 2 patients sustained 3 times of laparotomy respectively. Overall mortality rate was 12%(4/33). The main causes of death were MODS resulted from consumption coagulopathy and/or severe septic complication after reoperation. Conclusions The effective preventive steps to decrease the incidence of reoperation in patients with abdominal injuries include complete interventional exploration, right operational methods and sufficient drainage in initial laparotomy for a surgeon.

ObjectiveTo investigate the role of nuclear factor kappa B (NF-κB), epidermal growth factor (EGFR) and Mucin 1 (MUC1) in hepatolithiasis associated with intrahepatic cholangiocarcinoma. MethodsSpecimens were taken from 90 patients who underwent hepatectomies from August 1989 to June 2009 at the Second Affiliated Hospital of Sun Yat-sen University. The specimens were stained immunohistochemically for NF-κB, EGFR and MUC1. There were 33 patients who had hepatolithiasis associated with intrahepatic cholangiocarcinoma (the experiment group). 32 patients with hepatolithiasis served as the control group, and 25 patients with normal intrahepatic bile ducts taken at 1-2cm distal to benign hepatic neoplasm served as the blank group. The immunohistochemical staining were performed on tissue slices. Results NF-κB positive rate was 51.5％ (17/33), 25％(8/32) and 4％ 1/25) in the experiment group, the control group and the blank group respectively,P＜0. 01 ; EGFR positive rates were 57. 6％ (19/33), 31.3％ (10/32) and 0 (0/25) respectively,P＜0. 01; MUC1 positive rates were 54. 5％ (18/33), 28. 1 ％ (9/32), 0 (0/25) respectively,P＜0. 01. There were significant differences for high level expressions of EGFR and MUC1 among histopathologic grading, tumor invasion and metastasis. The survival rates of patients with EGFR and MUC1 expressed tumor were lower than of patients with non-expressed tumout (P＜0. 01). ConclusionsNF-κB is probably involved in the early stage of intrahepatic cholangiocarcinogenesis. Overexpressions of NF-κB, EGFR and MUC1 play coordinately and important roles during intrahepatic cholangiocarcinogenesis. High level expressions of EGFR and MUC1 are related to the malignant degree of cholangiocarcinoma and to worse prognosis.

[Objective] To design routine MAGE-3 derived MHC-I/MHC-II restricted peptide epitope, which containing CD4~+-CD8~+ T cell epitope peptides antigen. [Methods] The epitope peptides were made through computer simulation designing, and peptide epitopes qualification tests were performed after the synthesis of peptide antigen, ELISPOT and cell-toxic analysis were used to evaluate the proliferation ability and cytokine-release ability of peptide-stimulated T cell. [Results] The sequence of obtained MAGE-3 derived restricted epitope peptide was FITC-YEEYYPLIFLDNDQETMETSEEEEYEEYYPLIF, of which the purity ≥ 90% tested by high performance liquid chromatography. MAGE-3 epitope peptide antigen could induce T lymphocyte proliferation, and induce T lymphocyte to secret IFN-γ, which higher than that of the control group (49 vs. 6 spots/10~6, P≤0.05 ). MAGE-3 epitope peptide could induce cytotoxic T lymphocytes to cause 42% of MFC cell lysis rupture, higher than control group (P≤0.05). [Conclusion] CD4~+-CD8~+ T cell epitope MAGE-3 peptide antigen showed considerable immunological effect in vitro, and such a peptide antigen can work as therapeutic polypeptide vaccine for H-2K~K mice gastric cancer which express MAGE-3 antigen.

Objective To investigate the assessed value of tumor-associated macrophages ( TAMs ) and KIT expression for liver metastasis in pancreatic neuroendocrine tumors (PNETs) and patients′outcome. Methods A total of 79 patients who underwent surgical resection and pathologically diagnosed as PNETs in the Department of Hepatopancreatobiliary Surgery in Sun Yat-sen Memorial Hospital from January 1995 to May 2015 were enrolled.The immunohistochemical staining of CD68 and KIT were detected and the correlations with clinicopathological factors were analyzed.Results Of 79 PNETs cases, CD68 and KIT in tumor tissue were overexpressed in 30(38%) and 35(44.3%) cases, respectively.CD68 overexpression was associated with tumor infiltration ( P<0.001 ), AJCC stage 7 ( P<0.001 ), liver metastasis ( P<0.001 ) and early recurrence (P=0.019).Patients with low CD68 level had significantly better survival than those with high CD68 expression ( P=0.0002 ).KIT overexpression was correlated with WHO 2010 and AJCC stage 7 (P<0.001;P=0.002), nonfunctional status of the tumor (P=0.002) and liver metastasis (P=0.026). The survival period of patients with low KIT expression was greatly longer than those with high KIT level (P=0.0013).CD68 and KIT co-overexpression was observed in patients with tumor invasion (P<0.001), advanced WHO and AJCC stage (both P<0.001) and better prognostic survival (P=0.0057).Multivariate analysis showed that CD68 overexpression (HR:2.9;95%CI:1.16~7.23;P=0.033) was an independent prognostic factor for PNETs.Conclusions CD68 and KIT overexpression is correlated with advanced disease stage, higher risk for liver metastasis and worse survival.CD68 is an independent prognostic factor for PNETs.

Objective To evaluate perioperative portal hemodynamic alterations in cirrhotic patients undergoing subtotal splenectomy,podicled spleen remnant retroperitoneal transplantation plus lower esophagus transection in the treatment of portal hypertension.Method Forty patients with cirrhotic portal hypertension were randomly allocated into 2 groups:splenic transplantation group (n = 20),in which patients underwent subtotal splenectomy with pedicled remnant spleen retroperitoneal transplantation and cardia-esophageal devascularization and transection,and control group (n = 20),in which splenectomy and cardia-esophageal devascularization and transection were performed.The cross section area,blood velocity and flow and collateral circulation of portal parameters were comparatively evaluated by 3D DEC MRA,and the size of remnant spleen,blood flow and collateral circulation of retroperitoneal transplanted spleen were comparatively assessed.Results At 6-month after operation,the disappearance of esophageal-gastric varices in two groups was similar,and the cross section areas of main portal vein (MPV) in two groups all decreased postoperatively,in study group it was (1.81±0.73) cm~2 vs.(1.20±0.52) cm~2,P ＜ 0.01;in control group it was (1.78±0.52) cm~2 vs.(1.30±0.12) cm~2,p ＜0.01.The mean blood velocity of MPV decreased postoperatively,in study group it was (9.86±0.10) cm/s vs.(7.06±1.92) cm/s,P ＜0.01;In control group it was (10.0 ±0.6)cm/s vs.(8.2±2.4) cm/s,P ＜0.01.The mean blood flow velocity of MPV in study group was lower postoperatively than that in control group(P＜0.01).The mean blood flow volume of MPV decreased postoperatively from (15.0±1.9) ml/s to (10.5 ±2.7)ml/s,P ＜0.01 in study group;and from (14.9±2.1) ml/s to (11.6±2.1) ml/s,P ＜ 0.01 in control group.The mean blood flow volume of MPV in study group was lower postoperatively than that in control group(P＜0.05).A significant collateral formation was observed around the retroperitoneally translocated pedicled remnant spleen.Conclusions Compared with splenectomy,subtotal splenectomy,retroperitoneal translocation of the pedicled remnant speen helps to preserve splenic function as well as to increase retroperitoneal collateral formation which is conducive to further decreasing the portal veinous pressure.

ObjectiveTo review our experience in the diagnosis and surgical treatment of solid pseudopapillary neoplasm (SPN) of the pancreas which can be used as a reference for other doctors to avoid misdiagnosis and to provide a better treatment.MethodThe clinical,laboratory,radiological,pathological and operative data of 24 patients with SPN of the pancreas operated between February 2001 to December 2009 were collected and retrospectively analyzed.Results23 of 24 patients were female and the mean age was 31 years.The most common clinical presentations were vague abdominal pain and abdominal mass.In most cases,abdominal imaging showed a solid or a solid-cystic mass in the tail or head of pancreas.All patients received surgery.20 of 22 patients who received curative resection were alive with no evidence of tumour recurrence.One patient who had a R1 resection died 42 months after surgery.The remaining patient was alive after a second operation.ConclnsionsSPN of the pancreas is a tumour with low malignancy.A correct diagnosis of SPN of the pancreas is made on its clinical,radiological and histopathological characteristics.Radical surgical resection is the treatment of choice.For patients with an advanced disease,palliative resection is beneficial.

Objective To investigate the value of modified T staging system in the diagnosis and treatment of hilar cholangiocarcinoma (HCCA).Methods The clinical data of 95 patients with HCCA who were admitted to the Memorial Sun Yat-Sen Hospital from December 1995 to January 2010 were retrospectively analyzed.Based on the results of imaging examination,preoperative staging was determined according the modified T staging system.The prognosis of the patients in difference T stages were compared.The data were analyzed by using the chi-square test and Fisher exact test.The survival curve was drawn by Kaplan-Meier method and the survival rate was compared by using the Log-rank test.Results The diagnostic rates of ultrasound + magnetic resonance cholangiopancreatography (MRCP),ultrasound + computed tomography (CT) or spiral CT were 93％ (37/40) and 66％ (23/35),respectively.The diagnostic rates of ultrasound + CT or spiral CT and endoscopic retrograde cholangiopancreatography (ERCP),ultrasound + CT or spiral CT and MRCP were 14/15 and 15/15,respectively.Of the 95 patients,44 received operation (including 28 cases of radical resection and 16 cases of palliative resection),16 received exploratory laparotomy,and 35 received simple internal or external drainage.For patients in T1,T2 and T3 stages,the resection rates were 71％ (30/42),50％ (12/24) and 7％ (2/29),respectively,with significant differences (x2 =30.182,P ＜0.05).The negative rates of the resection margins of patients in T1 and T2 stages were 77％ (23/30) and 5/12,respectively,2 patients in T3 stage were found with tumor residuals at the resection margin.There was a significant difference in the radical resection rate among patients in different T stages (x2 =8.204,P ＜ 0.05).Of the 44 patients who received surgical treatment,30 (68％) received concomitant partial hepatectomy.The ratios of patients in T1 and T2 stages who received concomitant partial hepatectomy were 70％ (21/30) and 9/12,respectively,with no significant difference (x2 =0.101,P ＞ 0.05).Fourteen (32％) patients received tumor resection.The incidences of complications and perioperative mortalities were 53％ (16/30) and 10％ (3/30) for patients who received concomitant partial hepatectomy,and 5/14 and 1/14 for patients who received tumor resection,with no significant differences between the 2 groups (x2 =1.188,0.094,P ＞ 0.05).The median survival time of patients who received concomitant partial hepatectomy was 29 months,which was significantly longer than 19 months of patients who received tumor resection (x2 =11.317,P ＜0.05).Eighty-six patients were followed up,and the median time of follow up was 15.6 months (range,3-70 months).The 1-year cumulative survival rates of patients in T1,T2 and T3 stages were 73.8％,58.0％ and 9.2％,respectively,and the 3-year cumulative survival rates of patients in T1,T2 and T3 stages were 33.5％,12.1％ and 0,respectively.The median survival time of patients in T1,T2 and T3 stages were 24,16 and 7 months,respectively.The prognosis of patients was getting poor as the increase of the T stages (x2 =37.07,P ＜ 0.05).Conclusions The modified T-staging system is beneficial to preoperative evaluation of patients with HCCA.Concomitant partial hepatectomy could improve the radical resection rate and prolong the median survival time of HCCA patients.