Objective: To explore the brain damage effects of lysophasphatidyl choline(LPC) in rats with pancreatitis and explore the pathogenesis of pancreatic encephalopathy(PE).Methods: SD rats were divided into test group,control group one and control group two at random.Acute edematous pancreatits rat model was induced following Aho HJ method for test group and control group one,then test group rats were venously applied with LPC and control group one was injected saline through tail vein.Control group two was treated with venous injection LPC through the rat tails without operation.Horseradish peroxidase(HRP) was used as a tracer to detemine if the blood-brain barrier(BBB)was open 7～10 days after successive application of LPC.The extravasated tracer was showed by diaminobenzidine(DAB).Rat brain tissue sections were examined by ponceau stain and Luxol Fast Blue stain to determine whether the rat brains were demyelination at the same time.Results: The BBB permeability of the test groupLTU rats increased greatly and obvious demyelination was observed in test group rat brains while both control groups had basically intact BBB and scarce demyelination was observed in both control groups.A statistical difference existed between the test group and control groups.Conclusion:LPC definitely open the BBB of the rat pancreatitis and demyelinates the brain of pancreatitis rats thus LPC plays an important role in the pathogenesis of PE.

Radical resection is one of the important factors for improving the prognosis of patients with resectable carcinoma of head of the pancreas,carcinoma of the distal bile duct and periampullary carcinoma.In order to proceed with a R0 resection,there are many types of pancreaticoduodenectomy (PD) for pancreatic,biliary and periampullary carcinoma such as PD with lymphadenectomy.In this report,we described a PD with extended retroperitoneal lymphadenectomy (D2 +) for the adenocarcinoma of the distal bile duct.The case presented underscores the feasibility and safety of PD with D2 + lymphadenectomy.

Objective To explore the effect of lysolecithin choline (LPC) on blood-brain barrier(BBB) permeability in rats with acute pancreatitis. Methods Acute pancreatitis rat model was produced and rats were randomly divided into:(1)Test group-AP, rats received LPC by tail vein injection; (2)control group I, AP rats were given normal saline by tail vein injection; (3)control group II, sham operation without AP, but LPC was given by tail vein injection.Horseradish peroxidase(HRP) was used as a tracer to determine BBB permeability 7~10 days later.Results The test group showed local extravascular effusion of HRP, indicating that BBB permeability was markedly increased,while both control groups showed no apparent increase of BBB permeability,which were statistically significant(P

Objective: To discuss the treatment of pancreatic encephalopathy(PE).Methods: Recombinant human growth hormone (rhGH)(Saizen)were applied in patients of severe acute pancreatitis(SAP) with suspicious early PE that presented with mental disorders. rhGH was also used in combination with somatostatin as the therapeutic method for SAP and its complication.Dosage and administration: Saizen 4U was injected intramuscularly twice a day for 5～7 days.Results: In 7 patients of this group, all of them showed improvement in the aspect of mental dysfunction and the symptoms disappeared after 48～72 hours. In 13 SAP cases underwent combined application of rhGH and somatostatin, no PE was observed.Conclusion: Application of rhGH showed therapeutic effect on the early manifestations of PE. It also suggested that combined use of rhGH and somatostatin could decrease the occurance of PE.

Objective To investigate the role of miR-200b on gemcitabine induced epithelialmesenchymal transition (EMT) in pancreatic cancer cell line MiaPaCa-2.Methods Different concentrations of gemcitabine were used to induce MiaPaCa-2,and the concentration of 50％ cell proliferation inhibited (IC50) was applied to obtain drug-resistant MiaPaCa-2 cells.MiR-200b or nonsense small molecular fragments (negative control,NC) was transfected into MiaPaCa-2 cells by liposomes,then gemcitabine of IC50 was used to induce cells to obtain drug-resistant MiaPaCa-2 cells transfected with miR-200b or NC.The morphological characteristics of MiaPaCa-2 cells were observed by inverted microscope.Invasion of cells were detected by transwell chamber.The expression of miR-200b was measured by using real-time PCR.The expressions of Ecadherin,Vimentin,Zebl,Zeb2 proteins were determined by Western blot.Results After gemcitabine treatment,the cells' size gradually diminished,intercellular junctions decreased,pseudopodium increased,which presented the characteristics of mesenchymal morphology.The invaded cell number increased from (26 ± 3) to (85 ± 6),and the expression of Vimentin Zebl,Zeb2 was increased to (1.87 ± 0.17),(2.57 ±0.21),(5.24 ± 0.83) folds of the parent cells.The expression of miR-200b was decreased to (0.36 ± 0.01)folds of the parent cells,and the expression of E-cadherin was decreased to 0.47 ± 0.05 folds of the parent cells,while the invaded cell number of drug-resistant MiaPaCa-2 transfected with miR-200b was decreased to (42 ± 4),and the expression of Zebl,Zeb2 was decreased to (0.36 ± 0.07),(0.08 ± 0.01) folds of drugresistant MiaPaCa-2 transfected with NC.Conclusions The occurrence of EMT is observed in pancreatic cancer cell line MiaPaCa-2 during gemcitabine induction,and miR-200b down-regulation may be a possible mechanism.

Objective To investigate the expression and methylation status of HOXA7 gene in human pancreatic cancer cell lines, and to explore the relationship between them.Methods HOXA7 mRNA expression of human pancreatic cancer cell lines BxPC3, CFPAC1, PANC1 and SW1990was detected by RT PCR.Bisulfite sequencing PCR (BSP) and combined bisulfite restriction analysis (COBRA) was used to test promoter methylation status.All the cell lines were treated by 5-aza-2-deoxycytidine (5-aza-dC), and HOXA7mRNA expression, methylation status was detected before and after this treatment.Results HOXA7 mRNA was expressed in BxPC3, CFPAC1 and SW1990, while there was no expression of HOXA7 mRNA in PANC1.HOXA7 promoter methylation rates of CFPAC1, BxPC3, PANC1 and SW1990 were 93.16%, 90.65%,90.09% ,52.30%.HOXA7 promoter methylation rate of SW1990 was significantly lower than those in other 3cell lines ( P ＜0.01 ).After 5-aza-dC treatment, HOXA7 mRNA of PANC1 was expressed again, and HOXA7mRNA of BxPC3 was increasingly expressed;while the expression of HOXA7 mRNA in CFPAC1 and SW1990was not significantly changed after 5-aza-dC treatment.Conclusions The expression of HOXA7 mRNA in BxPC3 and PANC1 was closely correlated with promoter hypermethylation, while there was no obvious relation in CFPAC 1 and SW1990.

Objective To determine the methylation status and expression of Ras association domain family 1A (RASSF1A), and the possible effect between promoter aberrant methylation and pathogenesis of pancreatic cancer. Methods The methylation status of RASSF1A promoter CpG island (CGI) pancreatic cancer cell line BxPC3 was detected in 5 cases of normal pancreatic tissue and 13 pairs of pancreatic cancer tissues (tumor and peri-tumor) by using COBRA (combined bisulfite restriction analysis) and the methylation rate was calculated. The RASSF1A mRNA expression of BxPC3 was compared between pre- and post-treatment of the inhibitor of DNA methyltransferase (5-Aza-2-deoxycitydine, 5-Aza-dC). Results The average methylation rate of RASSF1A promoter CGI was 62.90% in BXPC3, 9.14% in normal pancreas, 53.79% in peri-tumors (TP), and 55.82% in tumors. The methylation rates in port-tumors and tumors were significantly increased when compared with that of normal pancreas (P < 0.01), while there was no significant difference between in peri-tumors and tumors (P > 0.05). After 5-Aza-dC treatting BxPC3 cells, the methylation rates decreased to 42.5% (P < 0. 05) and RASSF1A mRNA expression was enhanced. Conclusions Aberrant hypermethylation of RASSF1A promoter CGI could be considered as an early event in the process of pancreatic cancer and participates in the pathogenesis of pancreatic cancer.

Objective To study CT value in diagnosing cystic lymphangioma.Methods There were 14 cases with cystic lymphangioma confirmed by pathology.Male were 6 cases and female were 8 cases,ranged in age from 1～57 years old,CT scans were performed in all patients.CT findings by comparison to operation and pathology were studied.Results Patients were subdivided into head-neck 4 cases; body 3 cases; viscus 7 cases based on the location of the lesions.The density in all lesions were homogeneous except one case with bleeding,the CT value were ?10 HU.The diameter of the lesions were 5～15cm, the margin of the lesions were clear, the adjacent tissue were compressed. The lesions were cycle or similar cycle, the septum in some lesions and the wall of lesions were thin, the septum and wall could be partially enhanced. The histories of disease were 1～10 years, average 4.5?1.6 years. No pain in patients except one case with bleeding were found, 3 cases in body had just a little uncomfortable.Conclusion The location and extent of cystic lymphangioma can be detected by CT and it is of valuable in guiding clinical treatment.

Objective To investigate the diagnosis and surgical management of adult choledochal cyst.Methods The clinical data of 58 adult patients with congenital choledochal cyst who were admitted to the First Affiliated Hospital of Nanjing Medical University from January 1997 to December 2010 were retrospectively analyzed.All patients were diangosed by the B ultrasonography,computed tomography (CT),Magnetic resonance cholangiopancreatography (MRCP) and endoscopic retrograde cholangiopancreatography (ERCP). Surgical procedures were selected according to the diagnosis and Todani classification.All data were analyzed using the t test or chi-square test.Results The accurate rates of B sonography,CT,MRCP and ERCP were 78％ (45/58),92％ (23/25),9/9 and 5/5,respectively.Forty-one patients underwent complete excision of the cyst + hepaticojejunostomy (2 patients were converted from laparotomy due to abdominal adhesions),2 underwent resection of the cyst and involed hepatic segments + hepaticojejunostomy,8 underwent laparoscopic excision of the cyst + hepaticojejunostomy,1 underwent left hemihepatectomy,3 underwent pancreaticoduodenectomy ( including partial hepatectomy in 1 patient),2 underwent common bile duct exploration + cholecystectomy due to acute obstructive suppurative cholangitis,1 underwent external drainage of choledochal cyst due to advanced malignance.The mean operation time and postoperative duration of hospital stay of patients who received open and laparoscopic excision of the cyst and hepaticojejunostomy were (235 ± 70) minutes,(320 ± 50) minutes,and ( 10.0 ± 2.3 ) days,( 12.6 ±6.6) days,respectively,with significant differences between the 2 groups (t =3.157,2.162,P ＜ 0.05).The postoperative morbidities of patients who received open and laparoscopic excision of the cyst and hepaticojejunostomy were 18％ (7/39) and 3/8,respectively,with no significant difference (x2 =1.515,P ＞ 0.05 ).Canceration of the choledochal cyst was observed in 6 patients( 10％ ).No perioperative mortality was observed,and the operative complication rate was 24％ (14/58).The duration of the follow up ranged from 1 to 15 years,no severe long-term complications were observed in patients with benign lesions.Four of the 6 patients with malignancy died in 1 year after operation,the other 2 patients survived for 3 years and 5 years,respectively.Conclusions Abdominal B ultrasonography should be the first choice for diagnosing adult congenital choledochal cyst,while MRCP is the gold standard.Surgical intervention should be timely considered once diagnosed. Complete excision of the cyst combined with Roux-en-Y hepaticojejunostomy is the first choice of treatment.

ObjectiveTo investigate the methylation of the promoter region in miRNA in pancreatic cancer cell line PANC1 and normal pancreatic tissue,to discover the miRNA with hypermethylation associated with pancreatic cancer.MethodsThe genomic DNA of PANC1 and normal pancreatic tissue was extracted,and fractured by ultrasound.Methylation DNA fragments were obtained by 5-methyl of pyrimidine nucleoside antibodies and immunomagnetic beads.The hypermethylation miRNA differentially expressed between PANC1 and normal pancreatic tissue was selected by using methylation DNA chip.BSP ( bisulfite genomic sequencing PCR) and TA clone sequencing was performed for further validation.The genomic DNA of pancreatic cancer cell lines BXPC3,CFPAC1,PANC1 and SW1990 was extracted.The COBRA (combined bisulfite restriction analysis) was used to validate differentially expressed hypermethylation miRNA.ResultsEight differentially expressed hypermethylation miRNAs were screened from the DNA methylation chips,then five of them were selected for sequencing.The methylation status of miRNA-615,-663,-663b was significantly higher in the PANC1 than in normal tissues (60.6％ vs 7.6％,88.8％ vs 22.2％,94.4％ vs 13.0％ ) ; the methylation status of miRNA-675 was not significantly different between PANC1 and normal pancreatic tissue (76.0％ vs 100％ ).Due to large error in sequencing,miRNA1826 was excluded.The results of COBRA confirmed all the 4 miRNAs were highly methylated in PANC1 ; except for miRNA-675,other 3 miRNAs were highly methylated in BxPC,miRNA-663,miRNA-663b were highly methylated in CFPAC1,while miRNA-615,miRNA-663 were highly methylated in SW1990.ConclusionsHypermethylation miRNAs were differentially expressed between pancreatic cancer cell lines and normal pancreatic tissue,among them,highly methylated miRNA-663 was possibly associated with pancreatic cancer.