Objective To detect the hypermethylation of HOXB6 gene in gastric cancer tissue and investigate the relationship between the gene expression and the clinics.Methods seven types of gastric cancer cell line and 73 cases of gastric cancer biopsy tissues were measured by combined bisulfite restriction analysis method.The gene expression and the re-expression after demethylation were detected by RT-PCR.Results Of 7 cell lines examined, 4 showed hypermethylation.The methylating status of HOXB6 was found to be correlated with loss of expression,which was restored by 5-aza-2＇-deoxycytidine treatment.We also find hypermethylation in 20/73(27.40%)case of cancer tissues but no one in non cancer tissue.Conclusion There are close relationship between hypermethylation and gene silencing in the HOXB6 gene of gastric cancer,There are also a gene re-expression after its demethylation.Hypermethylation of HOXB6 gene can be a tumor marker for gastric cancer diagnosis,and the agent of demethylation maybe a new antineoplastic drug.

Objective To evaluate the significance of percutaneous transhepatic cholecystoscopy ( PTCCS) in managing cholecystolitheasis. Methods From September 1999 to November 2001, eighty - six patients with symptomatic gallstones were allocated into this study. At first percutaneous transhepatic gallbladder drainage ( PTGBD) was carried out, one week later the formed fistula was dilated by bouginage to 16-22Fr in diameter, thereafter cholecystoscope was inserted into gallbladder through the dilated fistula to extract the stones using basket or electrohydraulic lithotripsy (EHL). Results PTGBD was attempted successfully in 82 out of 86 cases, PTCCS was performed in 80 of them. The amount of stones ranged from 1 to 76, single stone in 28 patients, stones more than two in 52 , with stone size ranged from 5 to 32 mm, and stones greater than 15mm in 45 cases. The stones were removed only using basket in 28, and by EHL in 54 cases due to their large size. The overall stone removal rate was 97. 5 % (78/80) . Complication of peritonitis appeared in 4 cases (4. 7% ) requiring emergent surgical intervention. The follow - up period with an average of 16. 4 months, gallstones recurred in 2 cases (2.5%). Stones were assayed by intra - red spectrom-etry in 30 of 54 patients with EHL, cholesterol stone existed in 24 of them. Conclusion PTCCS is relatively a safe and effective procedure for those patients with high risk on surgical cholecystectomy, or unsuitable to receive laparoscopic cholecystectomy. If the patients were selected strictly along the rules of indication, the complication and stone recurrence can be avoided.

Objective To study on gastric varices bleeding treated by tissue adhesive, D-TH glue injection. Methods Fifty-nine consecutive cirrhotic patients with gastric varices were undergone endoscopic D-TH glue injection with sandwich approach. Among these 59 patients, bleeding was present in 46 patients (acute bleeding in 37, recent bleeding in 9) , no previous of gastric variceal bleeding in 13. Follow-up study on therapeutic results and complications were conducted. Results Ninety-six injections were performed in all patients with a mean of 1. 63 injections (1-2ml) per patient. Recurrent bleeding never occurred in 37 patients with acute gastric varices bleeding 72 hours after injection, and the rale of emergency endoscopic he mostasis was 100 per cent. Thirty (65. 2% ) of 46 patients with bleeding gastric varices were followed for 12 36 months, 24. 9 months on average. Recurrence of bleeding occurred in 2(6. 7% ) , 5 ( 16. 7% ) and 8 (26. 7% ) cases at 3 , 12, and 36 months respectively. The rate of survival was 86. 5 per cent. Besides fever occurred postoperatively in 2 cases, other severe complications never happened. Conclusion Endoscopic D-TH glue injection is a safe and effective method for treating gastric varices bleeding.

Objective To investigate the relationship between bile amylase (BA) elevation and biliary tract disease in patients with normal pancreaticobiliary junction (NPBJ). Methods The bile juice was collected from bile duct in 202 consecutive cases who underwent therapeutic endoscopic retragrade cholangiopancreatography (ERCP) for biliary diseases ( biliary bile group), and from gallbladder (GB) in 73 consecutive cases who underwent percutaneous transbepatic cholecystoscopy for extraction of GB stone ( GB bile group). In biliary bile group, in addition to the measurement of BA, the level of lipnse (n =68), bacteria culture ( n = 149 ), manometry of Oddi's sphincter ( n = 27 ) and bile duct ( n = 38) were also performed. In GB bile group, additional GB biopsy was taken in 31 cases. No patient with pancreaticobiliary maljunction, history of cholangiojejunostomy or post-endoscopic sphincterotomy was included in the study. Results In biliary bile group, BA level was elevated in 95 patients (47. 0% ), in which there was no significant difference between neoplastic and non-neoplastic cases ( 56. 9% vs. 43.7%, P > 0. 05 ), although BA was elevated in most patients with hiler cholangiocarcinoma (7/9). The level of BA was correlated with bile lipase (r =0. 561 ), but not with pressure of Oddi's sphincter or bile duct. No significant difference in positive rate of bile bacteria culture was detected between patients with normal BA level and those with elevated level. In GB bile group, BA level was elevated in 25 patients (34. 3% ), in which the frequency of GB epithelium dysplasia is 87.5%, which was significantly higher than that from patients with normal BA level ( P < 0. 001 ). Conclusion The patients with biliary tract disease and NPBJ have high incidence of reflux of pancreatic juice into bile duct. In patients with elevated BA level, there was no significant difference between incidences of neoplastic or non-neoplastic disease, while the frequency of GB epithelium dysplasia and hilar cholangiocarcinoma were higher than those from patients with normal BA level.

Objective To investigate the risk factors of recurrent common bile duct (CBD) stones after treatment with endoscopic retrograde cholangiopancreatography (ERCP). Methods A total of 802 patients with CBD stone and with complete follow-up data were recruited to the study, among which 297 patients were accompanied with gallstone, 222 patients having undergone cholecystectomy before ERCP and 283 having no gallstones. The CBD stone recurrent frequency and the possible risk factors were calculated with uni- and multi-variate logistic regression analysis. Results The patients were followed up at a mean duration of 84. 8 months, and CBD stone re-occurred in 92 (11.5%). Both uni- and multivariate analysis showed gallstones, CBD larger than 1.5 cm, endoscopic mechanical lithotrity (EML) and angle of bile duct less than 120° were risk factors of recurrence. Multivariate analysis showed cholesterol stones, cholangeal stricture or sphincter of Oddis dysfunction (SOD) were all risk factors for reccurrence. Univariate analysis revealed that history of cholecystectomy or Billroth Ⅱ gastrectomy, stones larger than 1.5 cm and multiple stones were risk factors of recurrence. Conclusion For those with CBD stones undergoing ERCP, gallstone,common bile larger than 1. 5cm, EML and the angle of CBD less than 120° are major risk factors of recurrence. History of cholecystectomy and Billroth Ⅱ gastrectomy, SOD, large CBD stone ( ≥1.5 cm), cholesterol stone and multiple stones are also associating risk factors.

Objective To investigate the value of biliary tumor markers for differential diagnosis of the benign and malignant biliary tract diseases.Methods Tumor markers (CA19-9,CEA and CA242) examination and bacterial culture were performed in a total of 160 patients,who underwent therapeutic endoscopic retrograde cholangiopancreatography (ERCP) for biliary diseases.Resuts There were significant differences between malignant group and benign group in bile and serum in the level of CA19-9,CEA and CA242 (P ＜0.05) ; Cut-off value,according to ROC curve,was 239 ku/l in CA19-9,40 ng/ml in CEA and 60 ku/ml in CA242,respectively.There were significant differences between the bile marker and the serum marker in sensitivity,accuracy,negative predicative value of CEA (P ＜ 0.05).No significant differences was found in specificity between the serum group and the bile group.There were significant differences in bile CA19-9 level between cholangiocarcinoma,pancreatic cancer,duodenal papilla carcinoma with carcinoma metastasizing to bile duct,and hepatocellular carcinoma (P ＜ 0.05).Both in benign group and malignant group,there were significant differences in CA19-9 level between infectious bile and noninfectious bile (P ＜ 0.05).Conclusion The level of CA19-9,CEA and CA242 in bile can be applied to differentiate benign and malignant biliary diseases.The bile tumor markers do not have advantage over serum tumor markers in specificity for diagnosis.Bile bacterial infection can result in the elevation of bile CA19-9 while it does not have impact on differential diagnosis.

Objective To explore the impact of pancreatobiliary reflux (PBR) in normal pancreatobiliary junction on gallbladder.Methods A total of 54 patients receiving cholecystectomy for gallbladder diseases underwent ultrasonography to evaluate the thickness of gallbladder wall,inner layer and gallbladder wall blood flow before operation.The bile juice was sampled during ERCP in 45 patients with common bile duct stone and during cholecystectomy in 9 patients to detect amylase level.All patients with normal pancreatobiliary junction enrolled in the study were assigned into PBR group (n =24) and controlled group (n =30) according to their bile amylase level.Resected gallbladder specimens were examined histopathologically and then tested for expression of COX2,Ki-67 and p53 immunohistochemically.Results PBR group included 20 cases of cholelithiasis and 4 gallbladder polyp,among which 23 were occult PBR (OPBR) and 1 high confluence of pancreatobiliary ducts (HCPBD),which was similar to pancreatobiliary maljunction (PBM) pathologically.The control group recruited 28 cases of cholelithiasis and 2 gallbladder polyp.There were no differences in frequency of inflammation,hyperplasia,metaplasia or expression of p53 between the two groups (p ＞ 0.05),while higher presence of dysplasia and higher expression of COX2 and Ki-67 were seen in PBR group (p ＜ 0.05).Conclusion In patients with OPBR,although hyperplasia and metaplasia in gallbladder epithelium were similar to those induced by cholelithiasis,dysplasia and active proliferation might relate to progress to malignancy.

Objective To explore the impact of basal sphincter of Oddi (SO) pressure on pancreatobiliary reflux (PBR).Methods A total of 120 consecutive patients who received therapeutic ERCP for biliary tract diseases with or without the history of EST were enrolled,and were assigned to normal basal SO pressure group (n =23),elevated basal pressure group (n =55) and EST group (n =24).Basal SO pressure,bile amylase (BA) and bacteria culture findings were compared between the three groups.Results There were no differences in positive rate of bacteria culture,SO pressure increase and BA between the normal basal SO pressure group and elevated basal pressure group.In the latter group,negative correlation was observed between SO pressure and BA.The BA value [median(quartile range)] in patients with common bile duct ≥ 15 mm [4270 (12 337)U/L] was significantly higher than that in patients with common bile duct ＜ 15 mm [279.5 (1370) U/L].Furthermore,significant difference in both the proportion of elevated BA patients (83.3％ vs.59.0％) (P ＜0.05) and the positive rate of bacteria culture (75.0％ vs.33.3％) was seen between the intact papilla patients in the former two groups and those in EST group (P ＜ 0.05).Conclusion There was no direct correlation between the elevated basal SO pressure and PBR,but marked increase in diameter of common bile duct can lead to a rise in BA.Moreover,although EST can bring about PBR and bile bacterial infection,it may facilitate the outflow of the refluent pancreatic juice by relieving cholestasis.

Objective To investigate the safety of cleaning and disinfection of duodenoscope used in endoscopic retrograde cholangiopancreatography (ERCP).Methods The duodenoscopes,which were used in Endoscopy Center of Digestive Disease Hospital,Heilongjiang Provincial Hospital from May 4 to May 31 2016,were cleaned manually and disinfected with sterilization powder.The samples from elevator and elevator channel were collected for bacterial culture to evaluate the safety of clinical application of duodenoscope.Results Among 60 high level disinfections,one was excluded because disinfectant concentration wasn't effective.Among 59 elevator samples no positive cultures were detected,while among 59 channel samples,4(6.8％) cultures were positive and all from the same duodenoscope,including 3 cases of Klebsiella pneumonia and 1 case of Pseudomonas aeruginosa.Conclusion Sterilization powder is effective for disinfection of duodenoscope used for ERCP.Residual organic material can't be excluded from elevator sample even if it is negative for culture.Duodenoscope with high level disinfection still has the risk of transmitted infection,and should be monitored regularly by culture and reprocessed in time.

ObjectiveTo investigate the migration of bone marrow stem cells (BMSCs) to the liver and liver repair in mice with acute liver injury treated with BMSC transplantation through four approaches. MethodsMale BALB/c mice were divided into groups A, B, C, D, E, and F, with 10 mice in each group. Groups A, B, C, and D were treated by transplantation, group E was used as the donor of BMSCs, and group F was used as the model of acute liver injury. CCL4/2-AFF was used to establish the model of acute liver injury. Mouse BMSCs were isolated, labeled with the red fluorescent dye PKH26, and then transplanted into the mice with acute liver injury through the portal vein (group A), the tail vein (group B), the abdominal cavity (group C), and the spleen (group D). The mice were sacrificed 2 weeks later. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and albumin (Alb) were measured. The pathology of liver tissue was observed to evaluate the migration of BMSCs to the liver and the degree of liver repair. The mice in group F were sacrificed on day 8 to measure the levels of ALT, AST, and Alb. The t test was used for comparison of continuous data between two groups, and one-way analysis of variance was used for comparison of continuous data between multiple groups. ResultsIn groups A, B, C, and D, transplanted BMSCs migrated to the liver under a microscope, and newly formed hepatocytes were observed on pathological images. There were significant differences in the levels of ALT, AST, and Alb between groups A, B, C, and D and group F (ALT: t=2.372, 2.473, 2.354, and 2.383, all P＜0.05; AST: t=2.534, 2.423, 2.437, and 2.643, all P＜0.05; Alb: t=2.336, 2.243, 2.373, and 2.352, all P＜0.05). ConclusionBMSCs can promote repair of the liver in mice with acute liver injury, and the degree of liver repair is not related to the transplantation approach.