Objective To evaluate the clinical effectiveness of percutaneous transhepatic gallbladder drainage introduced by bedside ordinary-probe-guidance with free-hand technique in severe cholecystitis. Methods 85 cases treated with the operation were observed and analyzed. Results Infectious bile was successfully drained in all cases without serious complications. 83 cases received satisfactory outcomes. 2 elderly patients died from MODS secondary severe infection. Conclusion Bedside ordinary-probe-guided free-hand percutaneous transhepatic gallbladder drainage is a safe and convenient treatment with minimal invasion for severe cholecystitis, which can be widely used in clinic.

Novel intraductal radiofrequency ablation for malignant biliary stricture indicated good therapeutic prospect.In traditional intraductal radiofrequency ablation,the radiofrequency catheter was placed through endoscopic retrograde cholangiography or percutaneous transhepatic cholangiogram under the supervision of X-ray.In February 2014,a patient with intrahepatic adenocarcinoma of the bile duct and malignant common hepatic duct stricture was admitted to the Shenzhen People's Hospital.The patient received cholecystectomy + choledocholithotomy + T tube drainage 3 months prior to the admission.The results of histopathological examination indicated reactive hyperplasia.The patient recovered uneventfully after the operation.One week before the admission,the patient had jaundice and umber urine and received choledochoscopy + common bile duct dilatation + histopathological examination + biliary stent placement.The results of pathological examination confirmed that the patient had intrahepatic adenocarcinoma of the bile duct.The patient received intraductal radiofrequency ablation through T-tube fistula under direct vision of choledochoscopy,and recovered well after the operation.Intraductal radiofrequency ablation under direct vision of choledochoscopy not only has the advantages of accurate positioning and real-time monitoring,but also facilitate cyclical repeat treatment.

Objective To evaluate the significance of intraoperative laparoscopic ultrasonography (IOUS) during hepatobiliary surgery. Methods Sixty-eight cases of hepatobiliary lithiasis (liver disease 23 and cholecystopathy with difficulty in management by preoperative estimation, 45 ) were examined by intraoperative laparoscopic ultrasonography. Results Two CBD stones and one Mirrizi syndrome were found in IOUS. The laparoscopic procedures were successful fulfilled in 68 cases under the guidance of sonography including fenestration on liver cyst, incision and drainage on liver abscess, resection and biopsy on space occupied liver lesion with unknown cause, resection of solitary space occupied mass in liver, destruction of unre-sectable liver cancer by microwave and injection of absolute alcohol and chemotherapy through portal vein catheterization, cholecystectomy in acute cholecystitis, chronic cholecystitis with chiledolithiasis, and broad pedicle polyps in gallbladder etc. Conclusion IOUS has the advantages of non-invasive, fast, repeatable, high accuracy and has important clinical significance in laparoscopic diagnosis and treatment.

Objective To investigate the clinical evaluation on endoscopic sequential therapy for senile ACST.Methods The 36 cases of senile ACST were collected and divided into two groups:emergency operation and endoscopic sequential therapy.The results of treatment were compared between two groups.Result Comparing with emergency operation, the mortality of endoscopic sequential therapy was reduced from 27.8%to 5.6%,the rate of complication from 61.1% to 16.6%,the periods of preoperative preparation was shorten from 12.6 hours to 7.5 hours,the periods of postoperative recovery from 6.5 days to 3.5 days,the intervals between two treatments from 43 days to 11 days,the periods of complete treatment from 45 days to 23 days.Conclusion Endoscopic sequential therapy was the ideal alternative for senile ACST,which is characterized by minimal injury,effectiveness and thoroughness.

Objective To summarize the experience of surgical management of postoperative intrahepatic duct stones and strictures with choledochoscope.Methods The clinical data of 1241 patients with intrahepatic duct stones and strictures who were admitted to the Shenzhen People's Hospital from January 2000 to December 2011 were retrospectively analyzed.All patients were divided into postoperative choledochoscopy group (652 patients were treated by choledochoscope through T-tube fistula) and conservative treatment group (589 patients were treated by cholangic and antibiotic drugs) according to treatment plan.The clinical data including stricture alleviation,incidence of complications and treatment efficacies of the 2 groups were compared using the chi-square test.Results In the postoperative choledochoscopy group,the intrahepatic duct stones of 630 patients were completely removed,with the removal rate of 96.63％ (630/652),which was significandy higher than 76.23％ (449/589) of the conservative treatment group (x2=113.407,P ＜ 0.05).A total of 1237 intrahepatic duct strictures were detected in the postoperative choledochoscopy group,including 698 cases of membranous stricture (the length of strictured bile duct ＜ 2 mm),529 cases of spool-shaped stricture (2 mm ≤ the length of strictured bile duct ≤5 mm),8 cases of long tube-shaped stricture (the length of strictured bile duct ＞ 5 mm),2 cases of crackshaped stricture.The alleviation rate of the stricture was 99.35％ (1229/1237).The conditions of 6 cases of long tube-shaped stricture and 2 cases of crack-shaped stricture were not alleviated.There were 986 cases of intrahepatic duct strictures in the conservative treatment group,and the stricture alleviation rate was 80.43％ (793/986).There was a significant difference in the stricture alleviation rate between the 2 groups (x2=238.994,P ＜ 0.05).Nine patients in the postoperative choledochoscopy group had severe complications.Five patients had intrahepatic duct hemorrhage with a volume of blood loss more than 1500 ml,and they were treated by balloon dilatation hemostasis.One patient with repeated intrahepatic duct bleeding received hepatectomy.Two patients with severe intrahepatic cholangitis received choledochoscopy.Two patients had incomplete formation of sinus and they received implantation of biliary stents under duodenoscope.Conclusion Standard surgical management with choledochoscope can effectively alleviate postoperative intrahepatic duct stones and strictures.

AIM: To investigate the role of epigenetic modification in Pdx-1 gene transcription and expression, and to compare the differences between epigenetic modifications of Pdx-1 gene promoter in various cell types of mice. METHODS: The promoter DNA methylation and histone modification status of Pdx-1 and MLH1 genes in NIT-1 cells, NIH3T3 cells and mouse embryonic stem cells were measured by chromatin immunoprecipitation-real time PCR method. The expression levels of these genes in the three cell lines were measured by real time RT-PCR. The relation between epigenetic modifications and gene expression was analyzed. RESULTS: (1) Compared to mES cells, there was lower DNA methylation and higher H3K4m3 modification levels in the promoter of Pdx-1 gene in NIT-1 cells (P<0.05). DNA methylation, H3 acetylation, H3K4m3 and H3K9m3 modification levels in the promoter of Pdx-1 gene in NIH3T3 cells were distinctly increased (P<0.05). (2) Pdx-1 gene transcription expressed only in NIT-1 cells. The Spearman's rho between Pdx-1 gene expression and DNA methylation (r=-0.802,P<0.01) was observed. The Pearson correlation between Pdx-1 gene expression and H3K4m3 modification (r=0.997,P<0.01) was also found. The Spearman's rho between Pdx-1 gene expression and H3K9m3 modification (r=-0.879,P<0.01) was observed. (3) No correlation between housekeeper MLH1 gene expression and epigenetic modification was found. CONCLUSION: DNA methylation, H3K4m3 and H3K9m3 modification coordinated participate to regulate and control the expression of Pdx-1 gene. It is of great significance to the differentiation of β cells from ES cells.

Objective To explore the clinical value of sequential endoscopic therapy(SET) in treating severe acute cholangitis (SAC) in the eldly patients. Methods 38 eldly SAC patients were divided into 2 groups:emergency operation group and SET group. The efficacies in the emergency and stable periods were compared between the two groups. SET consisted endoscopic nasobilliary drainge (ENBD)first; after the disease was controlled, endoscopic sphincterotomy (EST) and stone removing by a basket with netting on top were done, and followed by laparoscopic cholecystectomy (LC) in patients with cholelithiasis in the stable period. Results Compared with the emergency operation group, the mortality of SET group reduced from 21.1%to 5.3%,the rate of complications from 57.9% to 15.8%, the preoperative preparation time was shortened from 12.4 hours to 7.6 hours, postoperative recovery time from 6.5 days to 3.5 days,and the period of total treatment from 45 days to 23 days. Conclusions SET is an ideal alternative for eldly SAC patients,it has minimum injury, effectiveness and shorter treatment course.

Objective To investigate the effects of targeted treatment of the carboplatin-Fe@C nanocage-loaded chitosan nanoparticles (C-Fe@CN-CN) combining external magnetic field on rats with transplanted liver cancer.Methods Twenty-four model rats with transplanted liver cancer were established and divided into four groups randomly (n =6).Abdominal exposure was carried out through a midline incision,and a cannula was inserted into the hepatic artery and fixed.Group A:saline water was injected as control,group B:saline water with 10 mg/kg free carboplatin was given,group C:saline water with C-Fe@CN-CN (equivalent dose of free carboplatin 10 mg/kg) was injected in absence of magnetic field,group D:saline water with C-Fe@CN-CN (equivalent dose of free carboplatin 10 mg/kg) was injected in presence of magnetic field for 30 min.All the animals were sacrificed and abdominal exposure was done again after 7 days.After tumors were reselcted,tumor weight and volume was measured,the inhibiting rate of tumor weight was calculated.Tumor and liver tissues were examined for histological changes.Results The growth of tumor was significantly inhibited after therapy with different forms of carboplatin.There was significant difference in the tumor weight of A,B,C,D groups [(0.85±0.12) g,(0.61±0.10) g,(0.48±0.09) g,(0.33±0.06) g,P ＜ 0.05,respectively].The inhibiting rates of tumor weight of B,C,D groups were 28.9 ％,43.4 ％,61.7 ％ respectively.The inhibiting rate of D group was highest which was 1.1 times higher than that of B group.There was also significant difference in the tumor volume of A,B,C,D groups [(1.06±0.24) cm3,(0.72±0.10) cm3,(0.50±0.07) cm3,(0.28±0.05) cm3,P ＜ 0.05,respectively].The tumor volume of group A was largest which was 2.8 times larger than that of group D.In group D,tumor tissues from six rats presented severe necrosis,and nanoparticles were concentrated in the necrotic tissue.In group C,five rats presented middle necrosis,one rats presented severe necrosis.There was no concentration of nanoparticles in the necrotic tissue.In group B,four rats presented middle necrosis,two rats presented mild necrosis.In group A,six rats presented mild necrosis.Conclusion C-Fe@CN-C can significantly increase the therapeutic effects of carboplatin by hepatic artery injection combining with an external magnetic field on the tumor.

Background and purpose:Hepatitis B virus X protein (HBx) and hypoxia inducible factor-1α(HIF-1α) play key roles in hepatocarcinogenesis and the development of hepatocellular carcinoma. Positive correlation on the expression of these 2 proteins in hepatocellular carcinoma tissues has been found, whereas the underlying mechanisms have not been fully elucidated. This study focused on the role of HBx in regulating HIF-1α and the underlying mechanisms in hepatocellular carcinoma cells. Methods:The expression plasmids were transfected into Huh7 cells with LipofectemineTM 2000. Western blot analysis was applied to detect the expressions of HIF-1αand HIF-1β protein. The transcriptional activity of HIF-1α was detected by the commercial analysis kits. The mRNA levels of HIF-1αand its target genes, including vascular endothelial growth factor (VEGF) and multi-drug resistance gene 1 (MDR1), were detected by quantitative real-time PCR (qRT-PCR). Immunoprecipitation analysis was applied to detect the interaction of HIF-1α, HBx and protein von Hippel-Lindau (pVHL). Results:Huh7 cells transfected with HBx plasmid led to sharp increase of HIF-1αprotein and transcriptional activity, as well as the mRNA of VEGF and MDR1 (P<0.05). However, the mRNA level of HIF-1αwas not obviously changed after HBx transfection (P>0.05). Meanwhile, HBx also signiifcantly impaired the function of pVHL in mediating the degradation of HIF-1αby ubiquitin hydrolase. This finding was further confirmed by the immunoprecipitation analysis, which showed that HBx could directly bind to pVHL, but not to HIF-1α. Conclusion:HBx may inhibit the inter-activation between pVHL and HIF-1αthrough directly binding to pVHL, and thus enhance the stability and transcriptional activity of HIF-1α.

Objective To observed the expression of matrix metalloproteinase-9 in the early onset of severe acute pancreatitis associated with acute lung injury in rats and investigate its effection in lung injury.Methods Thirty-two healthy adult male SD rats were randomly divided into two groups:Control group (n =8),Severe acute pancreatitis group(n =24).Severe acute pancreatitis model was induced by retrograde inject the 4％ sodium taurocholate sodium taurocholate into the biliopancreatic duct of rats.The severe acute pancreatitis group was detected the rate of lung water content、arterial blood gas.myeloperoxidase,matrix metalloproteinase-9,histopathology of the pancreas and lung injury score under the light microscope at 3 hours,6 hours and 12 hours.The matrix metalloproteinase-9 expression was detected by immunohistochemical and the results of immunohistochemical were analysed by the Image-Pro Plus image analysis system.Control group was detected the relevant indicators at 12 hours.Results Successfully modeling,the expression of matrix metalloproteinase-9 gradually increased beginning at 3 hours,at twelve hours up to the highest value(P ＜ 0.05).The degree of lung injury,lung water content,myeloperoxidase activity,PaCO2 gradually increased(P ＜ 0.05),PaO2 decreased significantly P ＜ 0.05).Conclusions The high expression of matrix metalloproteinase-9 is important to the pathogenesis of severe acute pancreatitis associated with acute lung injury.