Objective To investigate the clinical significance of tumor related gene gankyrin expression in pancreatic cancer.Methods The immunohistochemical staining was performed to study the gankyrin expression in 62 pairs of pancreatic cancer and matched non-tumor tissues.The correlations ofgankyrin expression with pathological parameters and the influences on the prognosis were analyzed.Results Ggankyrin positivity in pancreatic cancer and matched non tumor tissue was 2.5％ (45/62)and 19.4％ (12/62),respectively.The positivity of gankyrin was significantly higher in pancreatic cancer compared with matched non-tumor tissues,and the differences were statistical significant (P ＜ 0.01).Gankyrin expression in pancreatic cancer tissues were significantly correlated with TNM stage,differentiation and metastasis (P ＜ 0.05),but not with age,gender or tumor location (P ＞ 0.05).The 3-year survival rates of patients with positive and negative gankyrin expression were 17.8 ％ (8/45)and 41.2％ (7/17),respectively.Patients with positive gankyrin expression had a significantly shorter survival period compared with these with negative expression in pancreatic cancer tissues (P =0.034).Conclusions Gankyrin was overexpressed in pancreatic cancer,which was correlated with poor prognosis.

Objective To construct the stably over-expressing hB7-H3 gene human pancreatic cancer cell line PANC1,and provide tools for further investigating the function of hB7-H3.Methods hB7-H3 gene fragment was inserted into lentiviral plasmid GV287 carrying GFP to construct recombinant hB7-H3-GV287 plasmid vector.293T cells were transfected,and the GFP expression was evaluated under fluorescence microscopy.Western Blot wasused to detect the expression of hB7-H3 protein.Lentiviral vectors were packaged and the titer was determined.The recombinant hB7-H3 expressed lentivirus was used to infect PANC1 cell.Flow cytometry was applied to detecte the percentage of GFP and hB7-H3 positive cells.Real-time PCR and Western Blot was used to verify the mRNA and protein expression of hB7-H3.Self-cyclizing GV287 plasmid served as negative control (NC).Results PCR amplified fragment of recombinant plasmid was around 1 368 bp,and no amplified production of NC plasmid was observed.The DNA sequencing of recombinant plasmid was completely consistent with the designed fragment,indicating that hB7 H3-GV287 plasmid was successfully constructed.293T cells transfected with recombinant plasmid expressed hB7-H3 protein,while those cells transfected with NC plasmid did not express hB7-H3 protein.The virus titer of lentiviral packaged recombinant hB7-H3 plasmid was 2 × 108 TU/ml.The percentage of hB7-H3 positive cells,hB7-H3 mRNA and protein expression in PANC1 cells infected with cells infected with hB7-H3 lentivirus was 94.3％,5.09 ± 0.24 and 2.85 ± 0.27,respectively,which was obviously higher than 18.5％,1.28 ± 0.53 and 0.44 ± 0.69 in cells infected with NC lentivirus,and the differences were statistically significant (P value ＜0.01).Conclusions A human pancreatic cancer cell line PANC1 stably over-expressing hB7-H3 was successfully constructed.

Objective to determine the basal levels of several pancreas-related endocrine hormones in patients with chronic pancreatitis.Methods according to inclusion and exclusion criteria,consecutive patients from February 2016 to August 2016 in Department of Gastroenterology,Changhai Hospital,Second Military Medical University and ten healthy control (matched for age and gender) were included.Basal levels of glucagon-like peptide 1,pancreatic polypeptide,Secretin,glucagon,somatostatin between groups of CP vs healthy control,CP with abnormal glycometabolism vs CP with normal glycometabolism and alcoholic CP vs non-alcoholic CP were compared.Results a total of 53 patients with chronic pancreatitis and 8 healthy subjects were included in this study.(1) CP vs healthy controls:the level of secretin in healthy control patients is significant lower than that in CP patients;(2) CP with abnormal glycometabolism vs CP with normal glycometabolism:the level of triglyceride and somatostatin is significant lower than that in CP patients;the prevalence of patients with chronic alcohol consumption and the level of glucagon-like peptide 1 in CP with abnormal glycometabolism is significant higher than that in CP with normal glycometabolism;(3) the prevalence of abnormal glycometabolism in alcoholic CP group is significant higher than that in non-alcoholic CP.The results above are all of statistical significance.Conclusions in addition to dysfunction of islets/3-cells,CP also easily affects the level of other pancreas-related hormones such as secretin,somatostatin and glucagon-like peptide 1.Otherwise,chronic alcohol consumption is also strongly related with abnormal glycometabolism,the mechanism deserves further researches.

Objective To investigate the effects and adverse events of Yiaisanjie soup combined with sustained release fluorouracil in treating pancreatic cancer.Methods From January 2012 to December 2015,the clinical data of 62 cases with pancreatic cancer in Deqing County People's Hospital were retrospectively analyzed,which were divided into combination group and control group.The combination group underwent the implantation of sustained release fluorouracil panicles under CT guidance and took Yiaisanjie soup once a day for at least 5 cycles of 3-week administration and then 1-week no administration.The control group was treated only by sustained fluorouracil panicle implantations.The blood samples at fasting were collected before and 1 week after treatment to detect serum tumor markers.The adverse events were recorded.The therapeutic effect for solid tumors was evaluated according to response evaluation criteria in solid tumors(RECIST).Results Before treatment,CEA,CA125,CA242,CA19-9 in 2 groups had no statistical differences.After treatment,these tumor markers obviously decreased in both groups compared with before treatment.Compared with the control group,the CEA,CA125,CA242,CA50 and CA19-9 in combination group decreased at a more significant degree,and the difference was statistical significant (P ＜ 0.05).The incidence of Ⅱ and Ⅲ degree thrombocytopenia,nausea,vomiting and diarrhea was significantly lower in the combination group than in the control group (12.12％ vs 34.48％,18.18％ vs 48.28％,9.09％ vs 41.38％,all P＜0.05).There were 10 cases with partial remission (PR),17 cases with stable disease (SD) and 6 cases with progressive disease (PD),and the total effective rate was 81.8％ (27/33) in the combination group.There were 5 cases with PR,15 cases with SD and 9 cases and PD,and the total effective rate was 69.0％ (20/29) in the control group.The difference was not statistically significant (x2 =1.390,P =0.119).Conclusions The therapeutic effect of Yiaisanjie soup combined with sustained release fluorouracil in treatment of pancreatic cancer is not obvious,but the combination can significantly reduce serum tumor markers and reduce the incidence of side effects.

Objective To summarize the etiology and clinical features of recurrent acute pancreatitis (RAP) in the recent l0 years in China.Methods Pubmed,Medline,EMbase,CNKI,Wan fang and VIP database were searched for relevant articles published from January 2001 to December 2016 using Pancreatitis,Recurrence and Etiology as term index words.Meta analysis was conducted by RevManS.3 software.Results A total of 16 studies involving 3 980 patients (RAP n =1 231,AP n =2 749) were included.The age and sex were not correlated with AP recurrence.Biliary diseases,alcohol,inappropriate diets were associated with AP recurrence,but these were not the factors influencing RAP.Hyperlipidemia was responsible for the occurrence of pancreatitis in 17.00％ of RAP and 10.20％ of AP,with statistically significant difference (P =0.002).Hyperlipidemia was an important risk factor influencing AP relapse.The percentage of SAP in AP and RAP patients was 16.83％ and 24.13％,respectively,and the complication rate was 11.43％ and 15.13％,and the percentage of jaundice was 28.20％ and 32.53％.Those in RAP were higher than those in AP,and the differences were statistically significant (all P ＜ 0.05).Conclusions Hyperlipidemia was the predominant risk factor for RAP in China,and the patients with RAP tend to progress into SAP,and have jaundice and complications.

Objective To evaluate the efficacy of minimally invasive technique by retroperitoneal approach in the treatment of severe acute pancreatitis with peripancreatic necrosis.Methods The clinical data of 10 patients with severe acute pancreatitis combined with peripancreatic necrosis who underwent retroperitoneal drainage by aspirating and placing tube under B ultrasound guidance on the basis of routine therapy in the First Affiliated Hospital of University of South China from January 2016 to September 2016 were analyzed.Results All of the 10 patients successfully underwent puncture.Drainage was good after puncture and tube placement in 7 patients.The amylase activity was normal within 3 ～ 7 days,and the symptoms of abdominal pain and bloating were alleviated in 2 ～ 4 days.Among them,3 cases had poor drainage after catheterization due to a large necrotic area,and the percutaneous nephroscope was performed to remove the necrotic tissue of the pancreas.Bacteria was detected in the drainage fluid of 7 patients.The complications of pneumothorax,hemothorax and organ injury were not found in all the patients.The average hospitalization time was 19.4 days (10 ～ 34 days).Conclusions Minimally invasive retroperitoneal approach is safe and effective in curing severe acute pancreatitis with peripancreatic necrosis with good efficacy,which is worth for clinical recommendation and application.

Objectives To detect the expression of serum high mobility group box-1 (HMGB1) and explore its changes in rats with acute necrotizing pancreatitis (ANP).Methods Intraperitoneal injection of 20％ L-arginine in the dosage of 250 mg/100 g twice every 1 hour was used to establish ANP rat model.Intraperitoneal injection of normal saline solution in equal volume was performed in control rats.Rats were sacrificed at 6 h,18 h,24 h,36 h,48 h,72 h and 96 h after injection.Blood samples were collected to detect serum amylase and HMGB1 level.Pancreatic tissue was collected for pathological examination.Realtime PCR was applied to detect the mRNA expression of HMGB1 in pancreatic tissue.Werstem blot was used to determine HMGB1 protein expression in pancreatic tissue.Results Serum amylase level began to increase at 6 h after modeling,reached the peak at 18 h [(5 070 ± 603) U/L] and returned to normal level after 48 h.Serum amylase activity at 6 h and 18 h in ANP group was much higher than that in control group (1 844 ± 181)U/L(P＜0.05).The expression of HMGB1 began to increase at 6 h,reached to the peak at 36 h [(288.5 ±42.1)μg/L],and then decreased gradually.HMGB1 expressions at each time point in ANP group were significantly higher than those in control group (31.6 ± 10.1) μg/L],and the differences were statistically significant (all P ＜ 0.05).Pathological scores in pancreatic tissues in ANP group were higher than those in control group 0.38 ± 0.52,and the differences were statistically significant (P ＜ 0.05).HMGB1 mRNA expressions at t 6 h,18 h,24 h,36 h,48 h,72 h and 96 h in ANP group were 1.23 ±0.25,2.60 ± 0.46,3.23 ± 0.34,4.77 ± 0.66,2.88 ± 0.56,2.05 ± 0.20,1.33 ± 0.28,which were significantly higher than those in control group 0.44 ± 0.09,and the relative expression of HMGB1 in ANP group at 36 h was significantly higher than those at other time points (all P ＜ 0.05).HMGB1 protein expression in pancreatic tissue in ANP group at 6 h,18 h,36 h,72 h were 1.14 ±0.02,1.15 ±0.01,1.22 ±0.01,1.22 ±0.04,which obviously higher than those in control group(1.0),and HMGB1 expression in ANP group at 36 h was higher than those at other time points (all P ＜ 0.05).Conclusions HMGB1 may participate in systematic inflammation as one of the late inflammatory mediators during ANP.

Objective To examine the expression and analyze the significance of autophagy-related gene microtubule-associated protein 1 light chain 3 (MAP1-LC3,LC3),p62 and lysosorne-associated membrane protein 2 (LAMP-2) in pancreatic tissues of mice with acute pancreatitis (AP).Methods Twenty mice were randomized into AP group and control group,and the number of mice was equal between two groups.AP group was intra-peritoneally injected by 20％ L-arginine solution (two injections of 4 g/kg body weight,every 1 h) in the dosage of 4 g/l kg twice every 1 hour to establish AP model,while control group was administered with equal volume of normal saline by intra-peritoneal injection.All the mice were euthanized at 24 hour after the last injection.Pancreatic histopathological changes were measured.In addition,the protein expressions of LC3,p62 and LAMP-2 were detected by Western blot.Results No obvious pathological changes were observed in control group.Pancreatic acinar edema,structure destruction,missing,the obvious widening of interlobular septum,small interlobular septum and acinar septum,and the necrosis of acinar cells at different degrees were observed in AP group.The pathological score for tissue edema,hemorrhage,necrosis and inflammation in AP group was 3.13 ± 0.50,2.83 ± 0.32,3.25 ± 0.46 and 3.16 ± 0.47,respectively,which was all 0 in control group.The differences were statistically significant between AP group and control group (P ＜ 0.01).In AP group,the ratio of LC3-Ⅱ/LC3-Ⅰ,p62 and LAMP-2 protein in pancreatic tissue were 1.16 ± 0.08,0.94 ± 0.04 and 0.35 ± 0.04,respectively,which were 0.24 ± 0.02,0.34 ± 0.03 and 0.95 ± 0.03 in control group.The ratio of LC3-Ⅱ/LC3-Ⅰ and p62 protein in pancreatic tissue in AP group were much higher than those in control group,while LAMP-2 in AP group was lower than that in control group,and there was statistically significant difference between two groups (all P ＜0.01).Conclusions Intraperitoneal injection of L-arginine could induce acute pancreatitis,and autophagy is impaired,which was associated with decreased LAMP-2 protein expression.

Objective To explore the clinical application of function-preserving pancreatic surgery for treating pancreatic neuroendocrine neoplasms (pNENs).Methods Twenty-eight patients with pNENs treated by the function-preserving pancreatic surgery at Anhui Province Hospital from January 2002 to December 2015 were analyzed retrospectively.Results Among the 28 patients, enucleation surgery was performed in 18 cases, distal pancreatectomy was performed in 8 cases, and middle pancreatectomy was performed in 2 patients.Total average operation time was (197±68) min, and the blood loss was (106±99) ml.The postoperative pancreatic fistula was observed in 7 patients, seroperitoneum was in 4 patients, incisional infection was in 1 patient, pulmonary infection was in 2 patients, and all the patients were cured by conservative treatment.There was no death during perioperative period.The postoperative hospital stay was (13±6)d.Analysis of pathological examination and clinical symptoms showed that 24 cases were diagnosed as functional pNENs (including 23 with insulinoma and 1 with gastrinoma), and 4 cases were as non-functional pNENs.The tumor grade G1 was categorized in 19 cases, G2 was in 9 cases.The TNM was I stage in all patients.There were no vascular or nerve invasion or lymph node metastasis.The follow-up period ranged from 4 to 144 months.2 patients were lost, and other patients were all alive.No diabetes or hypoglycemia occurred.Conclusions The function-preserving surgery for pNENs was safe and feasible, especially for well-differentiated functional pNENs.

Objective To analyze the application of clinicopathological features and LEF-1 in the diagnosis of solid pseudopapillary neoplasm of the pancreas (SPN).Methods Clinical and pathological data of 227 cases who were pathologically diagnosed as pancreatic SPN at Changhai Hospital from Jan 2000 to Dec 2015 were collected and analyzed.Immunochemical assay was used to detect the expression of LEF-1 in 132 cases of SPN, and the results were compared with β-catenin, which is most commonly used for diagnosing SPN.Results 81.9% of patients with SPN were female (186/227).Mean age at the onset was 34 years.Mean tumor size was 5.4 cm.48.5% tumors were localized in the pancreatic tail, and 33% in the head.46.3% tumors were cystic and solid, 42.3% were solid, and 11.4% were cystic.There were 2 cases of lymph node metastasis (0.9%), 15 cases of vascular tumor thrombus (6.6%), 14 cases of nerve invasion (6.2%), and 13 cases of adjacent organs invasion (5.7%) based on microscopic observations.Immunochemical analysis showed that 130 of 132 cases with SPN expressed LEF-1 with strong nuclear positivity, and the positivity rate was 98.5%.But no obvious expression of LEF-1 can be seen in normal pancreatic tissue and other pancreatic tumors.The specificity was 100%.The positivity of β-catenin expression in SPN was 96.6%(144/149), and β-catenin was positively expressed in only one case of acinar cell carcinoma.Tumors were completely removed by surgery in 165 cases, and the median follow-up was 51 months.By Oct 31, 2016, 162 patients (98.2%) survived, 5 had liver metastasis, and 1 had recurrence.Conclusions SPN is predominantly encountered in young female patients, and the clinical manifestations are not specific.LEF-1 can be used as a specific marker for the diagnosis and differentiation of SPN, which is more accurate than β-catenin.