Objective To detect the levels of neutrophil extracellular traps (NETs) in patients with antiphospholipid syndrome (APS) and to preliminarily explore its formation mechanism.Methods Plasma samples from 27 APS patients and 30 healthy controls were collected.The circulating free DNA (cf-DNA) in plasma was detected by the PicoGreen nucleic acid quantitative assay kit,and the concentration of citrulline histone 3 (CitH3) was analyzed by enzyme-linked immuno sorbent assay (ELISA).The association of cf-DNA/NETs with thrombotic events in APS patients was further analyzed.The neutrophils in healthy controls were separated by density gradient centrifugation and stimulated with anti-β2GPl/β2GPI complex (100 μg/mL) for 4 h,and the cf-DNA/NETs in the culture supernatant was determined.TLR-4 inhibitor-TAK242 (5 μmol/L) was further used to observe whether the stimulation of the anti-β2GPI/β2GPI complex on cells could be intervened.The differences between groups were analyzed by analysis of variance (ANOVA) or rank sum test,Sidak or Dunnett's test were used to compare the mean of multiple samples and the correlation between variables was analyzed by Spearman's correlation test.Results The concentration of cf-DNA/NETs and CitH3 were significantly increased in plasma of APS patients compared with that in healthy controls [175.7(70.6,205.7) ng/ml vs 29.8(7.6,115.7) ng/ml,Z=-3.654,P＜0.05;19.5(7.8,26.4) ng/ml vs 3.3(0.84,10.3) ng/ml,Z=-3.932,P＜0.05],and there was a significant positive correlation between the cf-DNA/NETs and CitH3 (r=0.447,P=0.019).In the APS group,there was no significant difference in cf-DNA/NETs between patients with arterial thrombosis and those with venous thrombosis [177.1(67.8,297.2) ng/ml vs 184.7(82.4,233.9) ng/ml,Z=-0.301,P=0.786],whereas cf-DNA/NETs in the patients who experienced a new thrombotic event in 1 month was significantly higher than those with a history of thrombosis [192.1(83.6,328.8) ng/ml vs 90.0(42.8,184.7) ng/ml,Z=-2.006,P=0.046].In vitro,anti-β2GPI/β2GPI complex (100 μg/ml) stimulated the release of cf-DNA/NETs from neutrophils,which was significantly increased compared with the control group (t=10.39,P＜0.05),while TAK242 significantly inhibited the stimulating effects of anti-β2GPI/β2GPI complex on cells (t=4.22,P＜0.05).Conclusion The level of cf-DNA/NETs in peripheral blood of APS patients is significantly increased,which may play an important role in APS thrombosis.Anti-β2GPI/β2GPI complex induces the formation of cf-DNA/NETs through TLR4 and participates in the pathological process of APS.

Objective To evaluate the diagnostic accuracy of contrast-enhanced harmonic EUS (CH-EUS)for pancreatic cancer.Methods Patients with pancreatic occupying lesion underwent CH-EUS with ultrasonic contrast medium Sonovue.Pathological diagnoses by EUS-FNA or surgery and the follow-up results were made as the final diagnosis to evaluate the accuracy of CH-EUS for pancreatic cancer.Character-istics of CH-EUS in different pancreatic tumors were analysed.Results A total of 76 patients were en-rolled,with an average age of 53. 1 ±14. 2.Thirty-five were finally diagnosed as having pancreatic cancer, 21 local mass-type pancreatitis,10 pancreatic neuroendocrine tumor,6 cystadenoma,4 intraductal papillary mucinous neoplasm.Tumor diameter was 3. 4 ±1. 4 cm and there were 18 less than 2 cm.The sensitivity and specificity,positive and negative predictive value of CH-EUS for pancreatic cancer was 97. 1%, 92. 9%,91. 7% and 97. 5% respectively.The sensitivity was 100% combined with FNA.Conclusion CH-EUS is safe,convenient for pancreatic cancer with high accuracy and can be used as an additional diag-nostic method to EUS-FNA.

Objective To evaluate technical efficacy,feasibility and safety of a fully covered self-expanding metal stent for EUS-guided transgastric pancreatic pseudocyst drainage. Methods Data of a total of 11 patients who received EUS-guided transgastric pancreatic pseudocyst drainage with a covered self-ex-panding metal stent at Changhai Hospital from September 2013 to May 2014 were retrospectively studied. The manipulative success rates,curative success rates and complication rates were evaluated. Results All 11 patients were treated by EUS-guided transgastric pancreatic pseudocyst drainage with fully covered self-ex-panding metal stents successfully,with success rate of 100%. Two patients developed infection and displace-ment occurred in 1 patient. There was no hemorrhage,perforation or death. Stents were removed in 7 pa-tients and the pseudocysts vanished. Conclusion Endosonography-guided transgastric pancreatic pseudocyst drainage using a fully covered self-expanding metal stent can be accomplished with high technical and clinical success rate and low rate of complications.

Objective To evaluate the safety and efficacy of direct celiac ganglion irradiation with 125I seeds for pain relief secondary to advanced pancreatic carcinoma (PC).Methods This study enrolled 23 consecutive patients who had moderate to severe pain resulting from advanced PC.All patients underwent EUS-guided direct celiac ganglion irradiation with 125I seeds.Follow-up was conducted at least once weekly until death.Blood parameters,Visual Analog Scale (VAS) score,mean analgesic consumption,and complications were evaluated during follow-up.Results All patients successfully underwent implantation at one attempt.The mean number of seeds implanted in the celiac ganglion per patient was 4 (range 2-6).Immediately after the procedure,pain relief and analgesic consumption showed no significant changes compared with preoperative values.Six patients (26％) reported pain exacerbation.Two weeks later,the VAS score and mean analgesic consumption were significantly less than preoperative values.No procedure-related deaths or major complications occurred.Conclusion EUS-guided direct celiac ganglion irradiation with 125I seeds can reduce the VAS score and analgesic drug consumption in patients with unresectable PC.

Objective To analyse the clinical characterization of intraductal papillary mucinous neoplasm of pancreas (IPMNs) and to improve the understanding of IPMNs.Methods Three hundred and sixty-one patients with IPMN at Shanghai Changhai Hospital between 1993 and 2012 were retrospectively analyzed.Results Among 361 cases of IPMNs,241 were men and 120 were female,with a female to male ratio of 2.6∶1.Mean age of the patients was 62 years old (16 ～ 87 years old).The duct type included the main duct in 102 patients (28.3％),a branch duct in 109 (30.2％),and mixed ducts in 150 (41.6％).The most common symptom at presentation is pancreatitis,which occur in 167 patients (46.3％).The sensitivity for the detection of IPMN was 80.2％ by ERCP,76.9％ by MRCP,63.9％ by CT,and 50.5％ by EUS.One hundred and twenty-nine patients (35.7％) were operated and diagnosed with 87 (67.2％) IPMA,21 (16.4％) IPMB,10(8.2％) IPMC(CIS) and 10 (8.2％) had invasive carcinomas.The 5-year survival rates of IPMA,IPMB,IPMC,and invasive carcinomas were 100％,100％,66.5％,and 44.7％,respectively.Conclusions IPMNs were fregrently occured is 60 years old,half of patients had acute pancreatitis history and the frequtly attack,mixed type is most anatomy type,and the outcome after surgery is good.

Objective To evaluate the safety and efficacy of therapeutic ERCP for patients above 90 years of age.Methods The data of 37 patients of above 90 years who underwent 42 ERCP procedures from January 2001 to December 2009 were studied retrospectively and compared with those of 152 matched patients ( 168 procedures) below 65 years old at a 1∶4 ratio for success rate and complications.Results The rate of complete success,partial success,and failure in observation group was 73.81％ (31/42),19.05％(8/42) and 2.38％ (1/42),respectively,which were similar (P ＞0.05) with those in control group,with complete success rate at 85.12％ ( 143/168),partial success rate at 12.50％ (21/168) and failure rate at 2.38％ (4/168).The rate of terminated operation in observation group (4.76％,2/42) was significantly higher than that of the control group (0.00％,0,P =0.039).The overall rate of complication in observation group was 7.14％ ( 3/42 ),slightly higher than that of the control group ( 6.55％,11/168,P ＞0.05 ).There was no significant difference between the two groups regarding the rates and severity of such complications as pancreatitis,hemorrhage and infection ( P ＞ 0.05 ).No perforation or death was observed.Conclusion Therapeutic ERCP for patients of 90 years or older is safe and effective.Adverse events related to chronic concomitant diseases need early detection and proper management.

Objective To investigate the safety and efficiency of 125I radioactive stents for advanced extra-hepatic cholangio-carcinoma.Methods We retrospectively reviewed data of 15 consecutive patients with advanced and un-resectable extra-hepatic cholangiocarcinoma,who were treated by radioactive stents.Postoperative complications,patency time of stents,and survival of the patients were assessed.Results Fifteen patients underwent 32 endoscopic sessions of radioactive stents placement.Successful operations were achieved in all patients,and there were no life-threatening complications including perforation,bleeding or bone marrow depression.The average patency time of radioactive stents was 117 ± 105 days (8-295 days).The actual radiation dose was 56.55 ± 17.42 Gy (7.86-82.48 Gy).The median survival time was 420 days (90-1175 days) and survival of 6 patients exceeded 12 months.Conclusion The 125I radioactive stent is safe and effective for patients with advanced unresectable extra-hepatic cholangio-carcinoma.

Objectives To explore the expression of the receptor for advanced glycation end products (RAGE) in pancreas and the serum concentration of RAGE in rats with acute necrotizing pancreatitis (ANP).Methods Sixty four male Spraque-Dawley rats were randomly divided into control group,ANP 6,18,24,36,48,72,96 h group with 8 rats in each group.The rat model of ANP was established by injecting 20％ L-arginine intraperitoneally at the dose of 250mg/100g body weight for twice at the interval of 1 h.Rats in control group were injected intraperitoneally with the same amount of saline.The pancreas samples were histologically examined by light microscope and scored.The amount of ascites,levels of serum amylase and RAGE was determined; Real-time PCR was performed to detect the expression of RAGE mRNA in pancreatic tissue.Results Pancreatic injuries aggravated with time.The amount of ascites increased from ( 1.98 ± 0.64) ml at 6h to (8.69 ±0.62)ml at 96 h.Serum amylase level began to increase at 6h after L-arginine intraperitoneal injection and reached the peak value at 18 h[(5069.88 ± 603.25 ) U/L],and returned to normal at 36 h.The serum concentration of RAGE and RAGE mRNA expression in pancreatic tissue were ( 18.33 ± 2.99) ng/ml and 0.41 ±0.13 in the control group.The corresponding values increased at 6 h in ANP group,which were (30.31 ±5.03) ng/ml and 1.57 ±0.19,and they were significantly higher than those in the control group (P ＜0.05) ; they reached the peak value at 24 h[( 105.41 ±21.31 ) ng/ml and 4.23 ±0.73],which were significantly higher than those in other ANP groups ( P ＜ 0.05 ) ; at 96 h they decreased to the lowest point [( 33.54 ± 6.96) ng/ml and 1.19 ± 0.19],which were still significantly higher than those in the control group (P ＜ 0.05 ).Conclusions The expression levels of RAGE in pancreatic tissues and serum level of RAGE increase within 36 h of ANP onset,then decrease gradually,but they are always higher than normal values.

ObjectiveTo evaluate the accuracy of contrast-enhanced harmonic EUS (CEH-EUS) by using a second-generation contrast medium in differential diagnosis of pancreatic occupying lesions.MethodsPatients with suspected pancreatic neoplasms or chronic pancreatitis were enrolled and underwent CEH-EUS by using ultrasonic contrast medium Sonovue.Cytological and/or histological diagnoses were made by EUS-FNA and the final follow-up results.Characteristics of enhancement of the target areas,such as enhancement sequence,time features,pattern grade and venous elution degree were investigated.ResultsA total of 23 patients were enrolled,in which 13 were diagnosed with FNA as having pancreatic cancer,7 with chronic pancreatitis,2 with intraductal papillary mucinous neoplasms and 1 with microcystic serous cystadenoma.The accuracy of CEH-EUS was 95.65％,which was significantly higher than that of conventional EUS (78.2％).Enhancement of pancreatic cancer by CEH-EUS was later than or simultaneous with the nearby tissues,with heterogeneous low enhancement or fulfillment defect areas,and early subsidence without obvious peak.Enhancement of benign pancreatic diseases was simultaneous with the surrounding tissues,with homogenous fulfillment and simultaneous subsidence.ConclusionCEH-EUS is safe,convenient and accurate in diagnosis of pancreatic occupying lesions and can be used as an additional diagnostic method to EUSFNA.

Objective To investigate the diagnostic value of the K-ras mutations in FNA samples for early detection of pancreatic cancer. Methods FNA samples of 27 patients with pancreatic cancers, 9 patients with other malignant tumors and 14 patients with non malignant pancreatic mass (NMPM) were collected. DNA was extracted, and K-ras gene was amplified through PNA-mediated PGR clamping, the products were sequenced to determine the mutation type. Results The positive rate of K-ras mutations in pancreatic cancers,other malignant tumors and NMPM were 88.9%, 44.4%, 35.7%. There was significant difference in K-ras gene mutations in FNA samples between pancreatic cancer and other malignant tumors ( P = 0. 013 ) and NMPM ( P = 0. 001 ). The sensitivity, specificity, positive predictive value, negative predictive value,accuracy of K-ras mutations in FNA samples of pancreatic cancers were 88.9%, 55.6%, 85.7%, 62.5%,80.6% when compared with other malignant tumors, and the difference between the two groups was significant (P =0. 013) ;Those were 88.9%, 64.3%, 82.8%, 75.0%, 80. 5% when compared with NMPM, and the difference between the two groups was significant ( P = 0. 001 ). When cytology of FNA samples and K-ras mutations was combined, the positive rate of pancreatic cancer was up to 96.3%. Conclusions The detection of K-ras mutations in EUS-FNA samples helped improve the positive diagnostic rate of pancreatic cancer.