Objective We used the DNA methylation microarrays to investigate the differential methylation genes and loci sites in Kashin-Beck disease (KBD),to study the relationship between DNA methylation and KBD pathogenesis.Methods Totally 12 KBD adults and 12 healthy adults were selected and peripheral blood samples were collected and DNA was extracted.Illumina 450K bead-chip was applied to detect methylation status in KBD and healthy controls.Aberrant hyper-methylated sites were filtrated according to the P value after correction and methylation differences,together with GenomeStudio soft.Screened genes were validated using bisulfite sequencing polymerase chain reaction (BSP) technology.Results A total of 484 948 loci sites were analyzed and compared,93 differential methylated loci were found by comparing KBD and normal people,including 34 hypermethylated sites and 59 hypomethylated sites.There were 50 genes corresponding to the loci,43 genes not reported in literature.According to gene ontology analysis,the genes were involved in the immune response,antigen processing,phosphate and phosphoric acid metabolism and phosphorylation and the process of metal ions in combination.However,in the verification test using BSP method,there was no significant difference in methylation rate in human leukocyte antigen (HLA)-DRB1 between the case and the control group (48％ vs 70％,x2 =3.688,P ＞ 0.05).Conclusions The high and low differentially methylated sites in peripheral blood DNA of KBD patients are significantly different from those of the health control.HLA-DRB1 locus is not significantly different between the BSP verification test and methylation chip.

Objective The paper proposed suggestions and measures for hospital scientific research management by statistical analyzing all the papers published from 2009 to 2013.Method By referring to research database of this hospital and by using Microsoft Excel,this research calculated the frequency distributions of different variables including quantity,publisher,disciplinary subject,and the author title as well as core author.Result 772 papers were published from 2009 to 2013,mainly in local periodicals.75.52％ of the title of first author are high-level or intermediate-level;86 core authors published 378 papers,account for 48.96％ of the total amount.Conclusion The number of research papers is increasing year by year.but the quantity and quality are still low.The research administorsshould take measures to encourage researchers to publish quality papers in periodicals with higher standards.

Objective To investigate the psychological experiences of young uremic patients during different periods waiting for renal donor .Methods The phenomenology study method was adopted to conduct the semi-structure interviews in young uremic patients waiting for renal donor .Results The four themes were summarized:(1) having hope at the early stage (0—6 months);(2) ineffective coping at the middle stage(7—12 months);(3)uncertainty at the late stage(13—24 months);(4)serious negative emo-tion(more than 2 years) .Conclusion The young uremic patients patients waiting for renal donors have complex and changeable psychological experience .Medical staff should provide the pertinent professional support for promoting the living quality of patients by aiming at the psychological characteristics in young uremic patients .

Objective To evaluate endoscopic retrograde cholangiopancreatography (ERCP) for diagnosis and management of biliary cast syndrome after orthotropic liver transplantation. Methods A total of 71 consecutive patients with abnormal liver function and MRCP findings after liver transplantation underwent ERCP for diagnosis and management. Their data were retrospectively reviewed. Results A total of 188 sessions of ERCP were carried out on the 71 patients, most of whom were found to have stenosis of anastomotic stoma and/or bile duct. Bile sludge was found and removed in all patients diagnosed within 3 months after liver transplantation, while pigmentoid stones were found and removed in patients diagnosed within 3-6 months and biliary casts in patients diagnosed at more than 6 months. Each patient underwent 2.6 sessions averagely. Biliary casts were formed at an average time of 22. 7 ± 15.6 months after transplantation. PostERCP complications included 2 cases of pancreatitis and 3 cholangitis, with an occurrence rate of 2. 6%(5/188), which were all controlled with conservative treatment. The follow-up data was available in 56 patients showing improvement in liver function after ERCP, among who 42 met the endoscopic criteria of cure,1 0 received second liver transplantation because of progressive sclerosing cholangitis and 4 died from diseases other than liver transplantation. Conclusion Therapeutic ERCP for the biliary cast syndrome after liver transplantation is feasible, safe and effective, and can be performed repeatedly with good short-term effect.

Objective To evaluate the safety and efficacy of a newly designed anti-reflux metallic stent (ARMS) for malignant extra-hepatic biliary obstruction. Methods A total of 23 patients with unresectable biliary malignancy in the middle or lower part of common bile duct underwent endoscopic retrograde cholangiopancreatography (ERCP) and ARMs placement. The success rate, early complications, stent patency and patients' survival were recorded. Results The ARMSs were successfully placed in all patients and no procedure-related complication was recorded. The average operation time was similar to that of self-expanding metallic stents (SEMs). In 22 patients completing the follow-up, the total serum bilirubin dropped to normal within one month in 20. ARMs dysfunction occurred in 6, including tumor in-growth in 1, overgrowth in 2, and stent dislodgement in 3. Other patients were free of biliary symptoms until death or at the end of follow-up. The average stent patency of ARMs was 14 months. Patency rates at 3, 6, and 12 months were 95%, 74% and 56%, respectively. The mean patient survival was 7. 9 months ( 1-14 months).Postoperative survival rates at 3,6 and 12 months were 91.0%, 81.3% and 17. 2%, respectively. Conclusion Endoscopic placement of novel ARMs in patients with extra-hepatic biliary tumors is feasible,safe and effective.

Background and purpose:Nir1 is a transmembrane receptor for chemokine (C-C motif) ligand 18. CCL18 speciifcally binds to Nir1 at the cellular membrane of breast cancer cells to exert its invasion and metastasis. However, the speciifc mechanism of Nir1 is not clear in glioma. This study probed the effect and mechanism of Nir1 in the invasion of glioma cells.Methods:Western blot was used to detect the expression of Nir1 in glioma cells. siRNA plasmid was used to transfect U251 cells. Western blot was used to analyze the expression of Nir1 and protein phosphorylation of Akt in the cells transfected by Nir1 plasmid.In vitro Matrigel invasion assay was used to detect the invasive ability in the cells that were transfected. F-actin polymerization assay was used to detect F-actin recognition ability in cells.Results:The expression of Nir1 was higher in all glioma cells. After transfection, the invasion of siNir1/U251 was obviously decreased than the SCR/U251, F-actin content was reduced compared to the control group. Akt phosphorylation experiment result showed that the protein phosphorylation of Akt was enhanced in control group cells CCL18 following stimulation. However, the existence of CCL18 would affect the phosphorylation of Akt in siNir1/U251.Conclusion:Nir1 is high expression in glioma cells, and Nir1 binding to chemokine CCL18 promotes glioma cells invasion and metastasis through regulation the phosphorylation of Akt and F-actin polymerization .

The Internet Public Opinion Monitoring and Early Warning System for medical and health industry was designed and implemented due to the frequent occurrence of Internet public opinion, which has all-directional data collecting and analyzing functions, including big data collection, near duplicate detection, spam filtration, key public opinion early warning, region identification and tendency analysis, and can thus provide evidence for relevant departments to take effective measures for the control of Internet public opinion.

Objective To evaluate the effect of endoplasmic reticulum stress in trophocytes,in patients with intrahepatic cholestasis of pregnancy (ICP).Methods Sixty-one pregnant women who were hospitalized in Women's Hospital,School of Medicine,Zhejiang University from January to December 2015 were recruited.Thirty-one women who were diagnosed as ICP were defined as the ICP group and 30 healthy pregnant women were defined as the control group.The localization and expression intensity of glucose regulated protein 78 (GRP-78) in placental tissues were detected by immunohistochemistry technique.Electronic microscope was used to observe ultra-microstructure change of the endoplasmic reticulum in trophocytes and cell line Swan71.Reverse transcription (RT)-PCR and western blot were used to investigate the expression of GRP-78 mRNA and protein in Swan 71 cell.Results (1) GRP-78 protein was mainly expressed in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts.The protein expression of GRP-78 in placentas of the ICP group (13.2±2.4) was significantly higher than that in the control group (7.8±1.3,P＜0.01).(2) The volume of endoplasmie reticulum did not increase and the microvilli developed well,with no swelling and no expansion of endoplasmic reticulum in the control group.In the ICP group,microvilli injury,endoplasmic reticulum edema were found;the volume of endoplasmic reticulum increased,with dilation,vacuolation and significant degranulation.After treated with 100 μmol/L cholyglycine for 24 hours,universal dilatation of the endoplasmic reticulum were seen in the Swan71 cells.(3) In Swan71 cells,cholylglycine displayed a concentration-dependent up-regulation on the expression of GRP-78.The expressions of GRP-78 mRNA in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.17,2.17±0.16,5.47±0.36,5.65 ± 0.82,respectively.The expression of GRP-78 protein in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.04,1.17±0.15,1.33±0.13,1.73±0.13,respectively.The expression of GRP-78 mRNA and protein in 100 and 50 μ mol/L cholylglycine experimental group were significantly higher than 0 μmol/L (all P＜0.01).Conclusion The obvious expansion of endoplasmic reticulum and the increased expression of GRP-78 in trophocytes indicated that endoplasmic reticulum stress of trophocytes may be involved in the pathogenesis of ICP.

Objective To investigate an effective endoscopic management of biliary anastomotic stric-tures (AS) following orthotopic liver transplantation (OLT) and to evaluate the factors which may effect the ontcome. Methods Sixty-five patients, who were diagnosed as AS 3 months after OLT, underwent ERCP. Af-ter adequate dilation of the narrowing bile ducts, plastic stents, as many as possible, were inserted across the strictures and kept in place for at least six months. Results A total of 90 consecutive endoscopic procedures were performed in 65 patients. Before stents placement, the strictures were dilated by catheter or balloon (di-ameter range: 6-10 mm), or not dilated, according to the status of the bile ducts. An average of 3 (ranging from 2 to 6) plastic stents were placed with mean total size of 22.8 F (range 14-42 F), and the stents were kept for 8. 0 months on average (range 0.2-37.8 months). Of 90 procedures of stents placement, 54 (60%) were followed by stents removal and cholangiography, which confirmed stricture resolution in 26 (48.1%). The stricture resolution rate was 81.0% (17/21) in patients who underwent balloon dilation followed by more than 3 stents (> 21 F) for at least 3 months. Stricture re-occurred in 3 patients after stents removal, in whom stents were kept less than six months. Conclusion Endoscopic sequential intervention is effective for post-OLT biliary strictures according to the stage and grade. Radical dilation with maximal stenting can lead to complete resolution of AS. To achieve better result, if possible, balloon dilatation followed by three or mere endoprothe-ses (of at least 21 F) sustaining for more than 6 months is necessary.

ObjectiveTo retrospectively investigate the clinical features of the biliary tract complieations after living donor liver transplantation (LDLT) and the efficacy of endoscopy.MethodsThe LDLT patients with hiliary eomplieations were given endoscopic retrograde eholangiopanereatography (ERCP) and endoscopic therapies were carried out according to the result of eholangiogram.ResultsTwenty-one patients,among whom 43.8% were at early postoperative stage,underwent 28 endoscopic procedures.Nineteen patients (90.4%) had biliary anastomotic strictures with angled malformation.Bile leakage was found in 9 patients (42.9%).The success rate of endoscopic therapy was 85.7% ,including nose-bile drainage in 5,stent placement in 10,balloon dilatation and/or multiple stents placement in 9 and bile collection drainage in 2.During follow-up,leak heal was confirmed in 3 patients and stricture resolution was achieved in another 2 eases. ConclusionBiliary complication is relatively common in the early stage after LDIX,and severe anastomotie stricture with angled malformation is most frequently seen, usually associated with bile leakage. Series of endoscopic interventions with the strategy of "leak first,and then stricture"might achieve satisfactory outcome.