AIM:To investigate the influence and mechanisms of human pancreatic cancer tumor microenvironments on T-cell immunoglobulin mucin-3 (TIM-3) expression and function of dendritic cells (DCs).METHODS:Tumor-infiltrating dendritic cells (TIDC) and para-carcinoma tissue DCs were isolated by Ficoll-Hypaque density centrifugation from trypsinized pancreatic carcinoma tissues, and the peripheral blood mononuclear cells were isolated from pancre-atic cancer patients or healthy people.The expression of TIM-3 on DCs was detected by flow cytometry.DCs isolated from healthy people peripheral blood mononuclear cells were induced by rhGM-CSF and IL-4.The expression of TIM-3 in the DCs treated with the culture supernatants of Capan-2, SW1990 and Panc-1 pancreatic cancer cells or human skin fibroblast (Hs27) cells for 48 h, and in the DCs treated with supernatants of Capan-2 cells, anti-VEGF-R2, anti-IL-10 and EP2 re-ceptor blocking peptide were evaluated by flow cytometry.The releases of IFN-βand IL-12 in the culture supernatants of DCs pretreated with monoclonal antibody ( mAb) to TIM-3 or DNase+RNase, followed by stimulation with apoptotic Ca-pan-2 cells, were detected by ELISA.RESULTS:DCs in tumor microenvironments had higher expression of TIM-3 than the DCs from para-carcinoma tissues and pancreatic cancer patient or healthy people peripheral blood ( P<0.01 ) .TIM-3 expression in the DCs treated with the culture supernatants of Capan-2, SW1990 and Panc-1 pancreatic cancer cells for 48 h was much higher than that in Hs27 cells (P<0.05).Treatment with a combination of anti-VEGF-R2, anti-IL-10 and EP2 receptor blocking peptide largely diminished the upregulation of TIM-3 on the DCs mediated by Capan-2 cell superna-tants (P<0.05).The concentrations of IFN-βand IL-12 in the DCs with high expression level of TIM-3 were lower than those in the DCs with low TIM-3 expression level.Treatment with mAb to TIM-3 resulted in much more IFN-βand IL-12 releases (P<0.01), but DNase+RNase made this effect disappear.CONCLUSION:TIM-3 serves as a negative regula-tor of DCs innate immune responses in the pancreatic cancer microenvironments.The secretion of soluble factors to tumor microenvironment by pancreatic cancer cells, including IL-10, VEGF and PGE2 , may contribute to the regulation of TIM-3 expression.

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 explore the role of interleukin (IL)-1β in the pathogenesis of gout.Methods Real-time quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay(ELISA) were used to measure the expression of IL-1β mRNA in peripheral blood mononuclear cells (PBMCs) and IL-1β in plasma samples from 120 acute gouty (AG) arthritis,70 chronic gouty (CG) arthritis,80 intercritical gouty (IG) arthritis patients and 96 healthy control subjects respectively.Results The expression of PBMCs IL-1β mRNA and plasma concentration of IL-1β were both much higher in gout patients than those in controls (P ＜ 0.01,respectively).And the plasma levels of IL-1β mRNA and IL-1β significantly increased in the AG group compared with CG and IG groups (P ＜ 0.01,respectively) and much higher in the CG group than those in the IG group.Positive correlations existed between plasma concentration of IL-1β and the levels of white blood cell,neutrophil,monocyte,erythrocyte sedimentation rate,blood uric acid,globulin and PBMCs IL-1β mRNA (P ＜ 0.01,respectively) while negative correlation between plasma IL-1β and plasma level of apolipoprotein in gout patients (P ＜ 0.05).Conclusion Elevated plasma level of IL-1β may be involved in the pathogenesis of acute and chronic gouty arthritis.

Objective To explore the value of anatomical liver resection in the treatment of bilateral intrahepatic biliary lithiasis. Methods We collected the clinical data of 32 patients with bilateral intrahepatic biliary lithiasis who received anatomical liver resection and 25 patients who recevied non-anatomical liver resection from May 2010 to May 2012 in our hospital and Sun Yat-sen Memorial Hospital. We comapred the therapeutic efficacy of these two operative modalities in the diagnosis and treatment of bilateral intrahepatic biliary lithiasis. Results The intraoperative blood loss was 436 ±48.162 mL in patients who received anatomical liver resection, and was significantly less than that in the control group (763 ± 37.645ml) ( < 0.05 ) . Postoperative liver function status:for patients who received anatomical liver resection,on the third day after operation,the total bilirubin and AST were significantly lower than those in the control group ( <0.05), and the postoperative hospitalization time was also shorter than that of the control group ( <0.05 ) . The two groups in the operation time, postoperative complications and residual calculus had no significant difference ( >0.05 ), but there 1 patients died of liver failure in the control group. Conclusions Anatomical liver resection is a favorable method to completely remove the lesions under the premise of retaining the residual liver function as much as possible. The rate of remnant biliary lithiasis and recurrence is lower and the recovery is quicker in these patients after anatomical liver resection. Thus, anatomical liver resection is worthy of promotion.

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 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 detect the variation of vascular endothelial growth factor (VEGF) in the splanchnic vessels under portal hypertension (PHT) and explore its mechanism and influence on the process of hyperdynamic circulation.Methods In humans,the level of VEGF pathway related proteins were detected in the splenic artery of PHT patients in clinical trials.In animal experiments,the following parameters were observed for rats in the control group (group N,n =7) and the CCl4 induced portal hypertension group (group PHT,n=7):portal vein diameter,portal vein blood flow velocity (PBV),portal vein blood flow (PBF),portal vein pressure (PP),norepinephrine (NE) reactivity in the isolated mesenteric artery microcirculation,contractile reactivity and degree of endothelial ni tric oxide synthase (eNOS) activation in the mesenteric artery by selectively inhibiting the VEGF signal pathway with SU5416.In cell experiments,primary culturing of arterial endothelial cells in vitro were used to verify the effects of VEGF on eNOS activation.Results The results showed that VEGF expression levels in the splenic artery of PHT patients significantly increased.In animal experiments,there was not a significant difference in portal vein diameter between group N and group PHT.How ever,the PBV and PBF of group PHT were lower than those of group N,and SU5416 had no clear effect on PBV and PBF in group PHT.PP of group PHT was much higher than that of group N,and SU5416 had little influence on reducing PP in group PHT.The contractile response of mesenteric artery microcirculation to NE in group PHT decreased significantly,EC50 increased a lot,and SU5416 improved this hypoergia partially.The protein levels of VEGF,VEGFR-2,eNOS,and p-eNOS in the mesentery artery of group PHT raised quite a lot compared to group N,and SU5416 decreased the protein level of VEGFR-2 and activation of eNOS significantly.Experiments in vitro confirmed that VEGF could promote the activation of eNOS.Conclusion The excessive VEGF produced in visceral arteries under PHT may participate in the process of hyperdynamic circulation partially through promoting the synthesis and activation of eNOS and then reducing visceral arteries' response to NE.

Objective To evaluate the value of radiofrequency ablation and Percutaneous Ethanol Injection in the treatment of small hepatocellular carcinoma (HCC). Methods We searched MED-LINE (1966-2009), EMBASE (1966-2009), CBMdisc (1978-2009), The Cochrane Library, Evidence Base Medicine Reviews (Ovid Edition), and Cancerlit (1993-2009). Date of last search: 30Jun 2009. There were no restrictions in language. Randomized controlled trials (RCTs) and nonRCTs (NRCT) were both included in this study, and the quality of each study included was assessed.Meta-analysis was performed using RevMan 4.2 software. Results Four RCTs and one NRCT met the inclusion criteria on RFA versus PEI in the treatment of small HCCs. Meta-analysis showed the following: complete tumor response rate, 3-year survival rate, 1-, 3-year tumor-free survival rates and 1-, 3-year local recurrence rates showed statistically significant difference in the RFA group than the PEI group(P＜0.05). The 1-year survival rate and the main complications of the two groups of patients were similar and they were not significantly different (P＞0. 055). Conclusions The results show that RFA resulted in better clinical outcomes than PEI in the treatment of small HCC larger than 2 cm, and no difference small HCC of 2 cm or less. The two modalities were safe and there were vey few adverse effects of the treatments.

Objective To investigate the role of miR-223 in the pathogenesis of acute gouty inflammation.Methods The subjects were divided into 3 groups:65 acute gout patients (AG),50 inter-critical gout patients (IG),and 45 healthy controls (HC).The peripheral blood mononuclear cells (PBMCs) and the clinical laboratory parameters were all collected.The expression of miR-223 in the PBMCs was detected using realtime fluorescent quantitative polymerase chain reaction (RT-qPCR) (TaqMan probe).The PBMCs of 5 healthy people were stimulated with monosodium urate (MSU) (100 μg/ml) for 12 h,and then,miR-223,NLRP3 mRNA and IL-1β production were all measured using RT-qPCR and ELISA respectively.All data were analyzed by SPSS 17.0 statistical software,Wilcoxon rank sum test,t test and Spearman's correlations analysis were used for statistical analysis.Results ① The expression of miR-223 in AG and IG groups was both significantly decreased than that in the HC group (8±17 vs 26±76,P＜0.05;9±17 vs 26±76,P＜0.05;respectively),AG group was significantly decreased than that in the IG group [8(17) vs 9(17),Z=11.387,P＜0.01].② After stimulated with MSU in healthy controls,IL-1β production and NLRP3 mRNA were both significantly increased [(86±5) pg/ml vs (13±6) pg/ml,t=21.042,P＜0.01;5.2±0.4 vs 1.2±0.4,t=14.640,P＜0.01;respectively],while the expression of miR-223 was significantly decreased (0.34±0.20 vs 1.05±0.24,t=-5.164,P＜0.01).Conclusion The data suggests that miR-223 might be involved in the patho-genesis of spontaneous regulation,but further study is needed to discover the exact mechanism.

Objective To explore the relationship of hepatocholangiocarcinoma and hepatolithiasis and to(increase) the early diagnosis of this disease.Methods The clinical data of 24 cases of(hepatocholangiocacinoma) associated with hepatolithiasis were retrospectively analyzed.Results The occurrence rate of hepatolithiasis concomitant with hepatocholangiocarcinoma was 2.8%(24 of 860 cases),and all of the tumors were located in the area of the bile duct with hepatolithiasis.Biliary atypical epithelial hyperplasia and cholangiocarcinoma were simultaneously present in some of the pathological sections.In this series,the correct diagnostic rate of hepatolithiasis associated with a space-occupying lesion in the liver before operation by ultrasonograph was 40.9%,by CT was 53.8% and by MRI/MRCP was 66.7%.17 cases(70.8%) were resected,and the radical resection rate was 33.3%(8/24).Palliative resection was done in 37.5%of cases(9/24).The 1-and 3-year survival rate was 62.5%,and(25.0)% respectively,in the group of radical resection;was 33.3% and 11.1% respectively,in the group with palliative resection;and 0% in the pathological biopsy group.Conclusions Long-term recurrent hepatolithiasis is an important cause for the development of hepatocholangiocarcinoma.The misdiagnostic rate of the disease was high,and the treatment results and prognosis were poor.It is important to pay more attention to the reasons for delaying the diagnosis of hepatolithiasis-related cholangiocarcinoma and take the appropriate preventative measures to assist in its early diagnosis.