Objective To explore the regulating mechanism of bcl 2 gene in hepatocyte injury caused by obstructive jaundice in rats. Methods Normal rats' and bile duct ligated 7d,14d,21d rats' hepatocytes were isolated by in situ collagenase perfusion and primary culture. (1) bcl 2 mRNA was detected by RT PCR in all cells; (2) After normal rat and bile duct ligated 14d rat hepatocytes were added 100?M GCDC and kept for 24hrs, cells were evaluated by FCM and TUNEL. Results (1) Normal rat hepatocytes did not express bcl 2 by RT PCR technique. bcl 2 was expressed in 7-,14-,21-day BDL rats. (2) After adding 100?M GCDC and keeping for 24hrs, the apoptosis of bile duct ligated rat hepatocytes significantly decreased compared with that of normal rat hepatocytes. Conclusions (1) Bile duct ligated rat hepatocytes expressed bcl 2. (2) Hepatocellular expression of bcl 2 during obstructive jaundice is an adaptive phenomenon to resist apoptosis by bile salts.

Objective To evaluate the difference of efficacy between continuous subcutaneous insulin infusion(CSII)and continuous intravenous insulin infusion(CVII)with human insulin in patients with diabetic ketoaciduria or ketoacidosis(DKA). Methods 120 patients with DKA were randomized into two groups,one was CSII group by a portable insulin pump(60 cases),the other was CVII group(60 cases).Results Both of CSII and CVII were effective in controlling blood glucose levels.CSII therapy provided better glycemic control(P

Objective To investigate the effects of epigenetic drugs on 2D- and 3D-cultured cholangiocarcinoma cells in vitro.Methods In this study,we have built compact and round TFK-1 spheroid in poly-HEMA coated 96-well plate and determined the effects of epigenetical drugs on 2D and 3D cultured cholangiocarcinoma cells:TFK-1.Viability of 2D and 3D cells model was examined by WST assay and FDA/PI staining. Using methylation-specific PCR analysis,we demonstrated the changes of methylation status of promoters regarding three tumor suppressor genes APC,E-Cadherin,and p16 INK4A.Results The average diameters of compact and round TFK-1 spheroids were in the range of 350-400 μm.The TFK-1 spheroid cells were more resistant to the epigenetic drugs and demonstrated a 11.2155-fold higher IC50 values for hydralazine and valproic acid than the same cells grown as monolayer. Higher doses of epigenetic drugs were needed to reverse the hypermethylation status in 3D cultured cells than 2D cells; however,the parallel dosage - dependent characteristic did not show in the 3D spheroid group.Conclusions Taken together,we established a 3D culture model of human cholangiocarcinoma epithelial spheroid.The 3D spheroid cells are more complex than the 2D monolayer cells and their unique characteristics are able to affect the consequences of epigenetic therapy.The 3D spheroid is a promising model for the research of epigenetic therapy.

Objective To study and evaluate islet cell adenoma of preliminary diagnosis.Method Retrospective analysis Wag done on 22 patients clinical data of preliminarily diagnosed islet cell adenoma.They were divided into two groups according to final diagnosis,group of islet cell adenoma(n=16)and group of reacfional hypoglycemia n=6).Results There were significant differences between the two groups in term of fames,cold sweat,cataphora,Whipple triad,hungry test,the ratio offasting blood insulin and fasting blood glucose exceeding 0.3 (I/G＞0.3) (P＜0.05).No statistical differences were found on blood glucose level in hypoglycemia onset,psychiatric symptom.Conclusion The clinic data is helpful for preliminary diagnosis,and Whipple triad,hungry test,I/G＞0.3 above all.

Objective To explore the regulating mechanism of inducible nitric oxide synthase(iNOS) in hepatic injury of obstructive jaundice (OJ) in vivo and in vitro experiments. Methods (1) Rat hepatocytes were isolated by in situ collagenase perfusion and primary culture. Hepatocytes were pretreated with various concentrations of iNOS inhibitor SMT for 20 min. After pretreatment, 50?M GCDC was added for an additional 24hr. Cells were next detected by FCM and TUNEL.(2) Experimental obstructive jaundice (BDL) was induced by double ligation of the bile duct in rats. After BDL for 3d、7d、14d、and 21d, the apoptotic status in liver of all rats were determined with TUNEL, and iNOS protein in liver of OJ was ditermined with immunohistochemistry method. Results (1) SMT decreased GCDC-induced apoptosis in a concentration-dependent manner. (2) The apoptotic rate of liver was related to length of time of OJ. Apoptosis index (AI) was highest from rats with 14d bile duct ligation. The stronger the iNOS expression, the higher was the number of apoptotic cells that was found in OJ. Conclusions iNOS is involved in the regulation and the occurrence and progression of hepatic injury of obstructive jaundice.

Objective To investigate the influence of duodenal diverticula on cannulation time and complication in patients undergoing ERCP.Methods Data of 3 265 patients undergoing ERCP in Drum Tower Hospital affiliated to Nanjing Medical University between January 1,2008 and December 31,2014 were enrolled.The patients' information and endoscopic pictures/videos were collected.The duodenal diverticula,cannulation time,postoperative complications were analyzed.Results There were 2 599 (79.6％) cases of non-diverticula,445(13.6％) cases of one-diverticula,122(3.7％) patients with two or more diverticula,and 99 (3.0％) intradiverticular papilla.Patients with duodenal diverticula accounted for 20.4％ (666/3 265) of all patients who received ERCP procedure.The mean cannulation time was 6.62 minutes in all cases,6.30 minutes in non-diverticula group,7.63 minutes in one-diverticula group,8.07 minutes in two-or-more group,8.58 minutes in intradiverticular papilla group,respectively.There were significant differences in cannulation time and complication rate between the groups.Conclusion Duodenal diverticula may be one of the factors that affect the cannulation time.It may enhance the cannulation complications and prolong the cannulation time,especially in those with intradiverticular papilla.

The effect of targeted magnetic nanoparticles on hepatoma and the underlying mechanism were examined. Nude mice transplanted with a human hepatoma cell line (HepG2 cells) were randomized into 5 groups, including: (1) group A, receiving normal saline, (2) group B, receiving 5-fluorouracil (5-Fu), (3) group C, receiving magnetic nanoparticles containing 5-Fu, (4) group D, consisting of treatment with magnetic nanoparticles containing 5-Fu and inside magnetic field and (5) group E, receiving pure magnetic nanoparticles and inside magnetic field. Morphological features of transplanted tumors in mice in each group were observed under transmission electron microscope (TEM). The expression of bcl-2/bax protein was immunohistochemically detected by SABC method. The results showed that a large number of apoptotic tumor cells were found in group B and group D under TEM. The expression of bcl-2 protein was significantly decreased and the expression of bax protein increased significantly in both group B and D as compared with those in group A, C and E (P<0.01 for all). The decrease in bcl-2 and the increase in bax were more in group D as compared with group B (P<0.01). It is concluded that the targeted magnetic nanoparticles containing 5-Fu can improve the chemotherapeutic effect of 5-Fu by decreasing bcl-2 expression, increasing bax expression and inducing apoptosis of the liver cancer cells.

The roles of protein kinase C (PKC) signal pathway in the pathogenesis of obstructive jaundice were studied. PKC from cytosolic and membrane fractions of peripheral blood lymphocytes (PBL) in 51 patients with obstructive jaundice and 16 cases of normal controls was isolated and purified. The activities of PKC were determined by radioactive isotope γ-32P-ATP-catalyzing assay. The results showed that the total PKC activities in PBL in the patients with obstructive jaundice were significantly increased as compared with those in the normal controls (P<0.01). Moreover, the membrane PKC activities and their percentages of the total PKC activities were higher in obstructive jaundice group than in those in the normal controls (P<0.05). The total PKC activities in PBL in the patients with obstructive jaundice were significantly positively correlated with the levels of soluble IL-2 receptor (sIL-2R) (r=0.58, P<0.01) and the degree of jaundice (T-BIL) (r=0.67, P<0.01) in serum. It was concluded that the activities of PKC signal pathway was related with the degree of T-BIL. PKC signal pathway might took part in the activation of T-lymphocytes in the patients with obstructive jaundice and play an important role in the immune regulation and the assessment of pathosis in the patients with obstructive jaundice.

The roles of protein kinase C (PKC) signal pathway in the pathogenesis of obstructive jaundice were studied. PKC from cytosolic and membrane fractions of peripheral blood lymphocytes (PBL) in 51 patients with obstructive jaundice and 16 cases of normal controls was isolated and purified. The activities of PKC were determined by radioactive isotope γ-32P-ATP-catalyzing assay. The results showed that the total PKC activities in PBL in the patients with obstructive jaundice were significantly increased as compared with those in the normal controls (P<0.01). Moreover, the membrane PKC activities and their percentages of the total PKC activities were higher in obstructive jaundice group than in those in the normal controls (P<0.05). The total PKC activities in PBL in the patients with obstructive jaundice were significantly positively correlated with the levels of soluble IL-2 receptor (sIL-2R) (r=0.58, P<0.01) and the degree of jaundice (T-BIL) (r=0.67, P<0.01) in serum. It was concluded that the activities of PKC signal pathway was related with the degree of T-BIL. PKC signal pathway might took part in the activation of T-lymphocytes in the patients with obstructive jaundice and play an important role in the immune regulation and the assessment of pathosis in the patients with obstructive jaundice.

Objective To investigate the risk factors of postoperative hemorrhage after pancreatoduodenectomy (PD).Methods The retrospective case-control study was adopted.The clinical data of 857 patients with pancreatic diseases who were admitted to the First Affiliated Hospital of Hunan Normal University from January 2007 to December 2014 were collected.All the 857 patients underwent PD and digestive tract reconstruction using the Child method.The number of patients with postoperative hemorrhage,classification,bleeding sites,source and time of bleeding and method and effect of treatment after PD were observed.The correlations among the gender,age,concomitant diseases (diabetes and hypertension),malignancy degree of tumor,the preoperative levels of serum alanine transaminase (ALT),total bilirubin (TBil),albumin (Alb) and prothrombin time (PT),international normalized ratio (INR),operation time,volume of intraoperative blood loss,method of pancreatic and jejunal anastomosis and postoperative hemorrhage after PD were analyzed.The follow-up of outpatient examination and telephone interview was performed to observe postoperative recovery of patients for 2 months till February 2015.Univariate analysis and multivariate analysis were done using the chisquare test and Logistic regression model,respectively.Results Of 72 patients with postoperative hemorrhage,grade A,B and C hemorrhage were detected in 3,41 and 28 patients,respectively,and 41,29 and 2 patients had respectively enteral hemorrhage,parenteral hemorrhage and enteral and parenteral hemorrhage.After PD,38 patients had hemorrhage located at the gastrointestinal tract,9 at the common hepatic artery,proper hepatic artery and gastroduodenal artery (5 due to pseudoaneurysm),5 at the pancreatic section,3 at the jejunal mesenteric vessels,2 at the middle colic arterial branches,1 at the superior mesenteric artery,1 at the superior mesenteric vein and 13 at the ambiguous bleeding sites.The early and late stage hemorrhages (within postoperative hour 24 and after postoperative hour 24) were detected in 20 and 52 patients,respectively.Of 44 patients with grade A and B of hemorrhages,17 underwent conservative treatment,16 underwent reoperation,8 underwent hemostatic therapy under gastroscopy,3 underwent interventional treatment.All the 44 patients had good hemostasis effect.Of 28 patients with grade C of hemorrhage,interventional treatment,reoperation,hemostatic therapy under gastroscopy,conservative treatment,interventional treatment + reoperation and gastroscopy + interventional treatment were applied to 10,7,4,3,3 and 1 patients,respectively.Ten of 28 patients died and 18 had successful hemostasis.The gender and preoperative levels of ALT and TBil were related factors affecting postoperative hemorrhage after PD in the univariate analysis (x2 =4.516,7.585,7.209,P ＜ 0.05).Male,preoperative ALT ≥ 172 U/L and preoperative TBil ≥ 159 μmol/L were the independent risk factors affecting postoperative hemorrhage after PD in the multivariate analysis (HR =2.033,1.860,1.872,95％ confidence interval:1.237-3.341,1.135-3.047,1.060-3.307,P ＜ 0.05).Fifty of 62 patients were followed up for a median time of 2 months with a follow-up rate of 80.6％ (50/62),and no rehemorrhage was occurred.Conclusion Male,preoperative ALT≥172 U/L and preoperative TBil≥≥ 159 μmol/L are the independent risk factors affecting postoperative hemorrhage after PD.