Objective To investigate the expressions of Yes-associated protein-1 (YAP1) in gallbladder mucosal epithelium of normal persons,in patients with simple/calculous cholecystitis,and in patients with gallbladder carcinoma;and to study the mechanism of YAP1 in gallbladder carcinoma development.Methods Immunohistochemistry was used to detect the expression and distribution of YAP1 protein in 50 persons with normal gallbladder,101 patients with simple cholecystitis/calculous cholecystitis and 100 patients with gallbladder carcinoma.RT-PCR and western-blot were used to detect the mRNA and protein levels of YAP1 in normal and malignant gallbladder mucosal epithelium cells.siRNA was used to shut down the expression of YAP1 in SGC996 cells.MTT was used to test cell vitality.Flow cytometry was used to measure cell cycle.Results Immunohistochemistry revealed the expression rates of YAP1 in the gallbladder carcinoma group,the cholecystitis/gallstone group and the control group to be 87.0％ (87/100),56.4％ (57/101) and 5.0％ (1/20),respectively (P ＜ 0.01).The YAP1 protein levels were higher in gallbladder carcinoma tissues and cells when compared to normal tissues and cells.RT-PCR showed the mRNA levels of gallbladder carcinoma cells to be 12.5 ± 1.2 times of normal gallbladder mucosal epithelial cells (P ＜ 0.05).After using siRNA to shut down the YAP1 expression,EMT associated proteins were down-regulated,cell vitality was decreased,and cell cycle was arrested in the S-phase.Conclusions YAP1 is closely related to cell proliferation and metastasis of gallbladder carcinoma.It may promote tumor progression through epithelial-mesenchymal transition.transition;Tumor progress

Objective By reviewing the relevant literature, we explored the current status and problems of the evaluation of the training effectiveness, and put forward the corresponding countermeasures, in order to provide a theoretical basis for the effect evaluation of nurse training in China. Methods By searching Citation online search and integration system (CITE), the Chinese biomedical literature service system (CBM), we included the literatures on the effect evaluation of the nurses training in the core journals from January 2006 to November 2015. Retrieval literatures were imported the bibliographic management software Note express by database. The corresponding field were analyzed in the literature′s age distribution, evaluation object, evaluation model, evaluation method, evaluation index and evaluation tool, et al. Results There were 343 literatures had retrieved, the total number of literatures presented a increasing tendency. The evaluation methods, evaluation tools and evaluation indicators presented a tendency of diversity, and the evaluation model was relatively single. Only 16 (4.7%) literatures tried to construct the training effect evaluation system of nurses, which was not based on some theory or hypothesis, which had not been effectively put into practice. Conclusion The effect evaluation system of nurse training needed to be done in depth, such as reconstruction of the new training evaluation model, the establishment of training evaluation system of different professional, different levels, different positions, to research evaluation methods, evaluation criteria and clinical practice testing, to design, training evaluation management software etc.

Objective To investigate the patterns for the recurrent and metastatic gastric cancer after curative resection and to indicate the strategy of treatment. Methods 162 patients who had received radical resection and presented post-operation failure during recent 9 years were analyzed. The failure patterns were confirmed by type-B ultrasonic or CT / MRI imaging. 15 of 34 patients with abdominal dropsy were diagnosed by adenocarcinoma cells in the abdominal dropsy. All superficial lymphadens and abdominal wall metastasis were diagnosed via punctuation. 31 patients with gastric remnant and (or) anastomoses recurrence were diagnosed via biopsy. Results Of 162 patients, 63 presented the recurrence or metastasis in multiple sites,including abdominal lymph node (LN) metastases in 46.9 % (76/162), peritoneum metastases in 21.0 % (34/162),gastric remnant and (or) anastomoses recurrence in 19.1% (31/162), liver metastases in 19.1% (31/162), the incidence rates of other parts were all ＜10 %. Meanwhile, out of 76 patients with abdominal lymph node metastases, 48.7 % (37/76) patients with peri-gastric LNs metastases, 31.6 % (24/76) with peri-pancreatic LNs metastases, 19.7 % (15/76) with para-aortic LNs metastases. The incidence of LN metastases was 57.7 % (56/97) in cancer arising from gastric fundus/cardia and 60.4% (29/48) in gastric body and 64.7 % (11/17) in pylorus antrum. Conclusion The regional failure sites for gastric cancer patients with radical resection were dominantly found in the gastric stump/stoma, the peritoneum and pelvic cavity implantation and abdominal cavity LN metastases, especially in the peri-gastric, peri-pancreatic and/or para-aortic LN metastases. The distant place failure sites were mainly in the liver, lung, brain, spondylus, cervical part LN and mediastinal LN metastases. Therefore, we should take chemotherapy, abdominal cavity chemotherapy and regional radiotherapy to prevent the regional district recurrences and distant metastasis after the gastric cancer patients with radical resection. Radiotherapy fields should focus on the gastric stump/stoma and the peri-gastric, peri-pancreatic and para-aortic LN regions.

Objective To explore the prognostic value of modified Glasgow prognostic score (mGPS) and risk factors in predicting overall survival (OS) in the castrate-resistant prostate cancer (CRPC) treated with docetaxel-based chemotherapy.Methods We retrospectively analyzed the data of 48 consecutive Chinese patients with CRPC received docetaxel-based chemotherapy in our institution from January 2008 to January 2012.Patients were divided into three groups according to the mGPS:0,1 and 2 score groups,and compare the OS among the three groups.Variables that were influenced the efficacy of chemotherapy were included in the univariate analysis and multivariate model.Survival analysis was performed using Kaplan-Meier curves,and the differences in overall survival rates were assessed using the Logrank test.Results The follow-up was performed until April 2014.There were 48 CRPC patients including mGPS 0 score group 30 cases,mGPS 1 score group 11 cases and mGPS 2 score 7 cases.The median OS was 22,11,9 months,respectively,P＜0.01.Univariate analysis showed that:prechemotherapy baseline total prostate-specific antigen (tPSA),Eastern Cooperative Oncology Group (ECOG) score,the number of chemotherapy cycle,visceral metastasis and PSA response were associated with poor prognosis (P＜0.05).Multivariate analysis showed that:prechemotherapy mGPS 1-2 score,baseline tPSA＞60 μg/L,the number of cycles of chemotherapy≤5,with visceral metastasis and PSA response in patients with CRPC were independent risk factors for prognosis in the CRPC treated with docetaxel-based chemotherapy.Conclusion mGPS is an independent risk factor for prognosis in the CRPC patients treated with docetaxel-based chemotherapy.

Objective To evaluate the effect of acupressure on chemotherapy-induced digestive tract reaction of malignant tumor patients.Methods We searched Pubmed,Medline,Embase,AMED,Cochrane Library,Cancerlit and Cinabl,China Biology Medicine disc (CBM),China National Knowledge Infrastructure (CNKI),VIP database (VIP)and Wanfang database,the databases were searched from the establishment of the database to March 2014.We identified randomized controlled trials (RCTs) and of acupressure on chemotherapy-induced digestive tract reaction for malignant tumor patients.Results In total,8 RCT,959 patients were included.Meta-analysis showed that compared with the no implementation of acupressure,acupressure could effectively reduce the frequency,duration and severity of chemotherapy-induced nausea,vomiting and retching.Meanwhile,acupressure could decrease anti-emetic medication dosage,but could not increase quality of life.Conclusions Acupressure can improve the digestive tract reaction of malignant tumor patients,alleviate the suffering of patients.Meanwhile acupressure couldn't increase the medical burden and mental burden of patients.More attention should be paid to the effect of acupressure on baseline level of state anxiety,functional state and more items.

Objective To investigate the levels of serum Golgi protein(GP73) (sGP73) in patients with HBV-related liver disease and investigate the role of sGP73 as an indicator for diagnosis of hepatocelluar carcinoma (HCC).Methods The concentration of sGP73 in patients with chronic hepatitis B (CHB,n =31),liver cirrhosis (LC,n =60),HCC (n =71),self-limited HBV infectors (n=21 ) and healthy controls (n =42) were tested by enzyme-linked immunosorbent assay (ELISA) assay and statistically analyzed in combination with relevant clinical indicators.Results The median of sGP73 in HBV-related liver disease group was significantly higher than that in the groups of self-limited HBV infectors and healthy controls respectively (P＜0.01).Among the groups with HBV-related liver disease,the median of sGP73 in LC group (231.13 ng/ml) was significantly higher than that in HCC without treatment group ( 117.63 ng/ml) (P ＜ 0.01 ) and CHB group (93.09 ng/ml) (P＜0.01).No significant difference was shown between HCC (without treatment) group and CHB group (P＞ 0.05),neither between self limited HBV infectors and healthy controls (respectively,36.79 ng/ml and 45.40 ng/ml) (P ＞ 0.05). The median of sGP73 in post-operation group (175.12 ng/ml,n=52) was significantly higher than in pre-operation HCC group (107.28 ng/ml,n=52) (P＜0.01).Along with the decreasing of liver function,sGP73 level was elevated in groups with HCC or LC.The receiver operating curve (ROC) constructed with the ratio of AFP and GP73 (AFP/GP73) showed a sensitivity of 78.87 ％ and specificity of 86.21 ％ with an area under the receiver operating curve (AUROC) of 0.878 (95％ CI:0.817-0.938) for diagnosis of HCC; comparably,a sensitivity of 67.61％ and specificity of 85.12％ were shown with a AUROC of 0.826 (95％ CI:0.755-0.897) when performed with AFP.Conclusion The level of sGP73 in HBV-related liver disease group is higher than that in the groups of self-limited HBV infector and healthy control,and it is positively correlated with the degree of hepatic impairment.For the diagnosis of HCC,joint detection of AFP and GP73 could achieve a better combination of sensitivity and specificity than the independent AFP test.

Abstract: Objective To express and purify MPT83 protein of Mycobacterium tuberculosis and evaluate its application value in immunological diagnosis of tuberculosis (TB) using clinical samples. Methods Using Mycobacterium tuberculosis (Mtb) H37Rv genome as the template, Mtb mpt83 gene was amplified by PCR and connected to PET-21a (+) to construct prokaryotic expression vector, and then transferred into E.coli DH5α. The positive colonies were picked out and retained. The recombinant plasmid pET-mpt83 of the strain with positive colony PCR was extracted, identified by double digestion, and the samples of the positive colonies were sent for sequencing. The correctly sequenced plasmids then were transferred into BL21 competent cells for induction, expression and purification with nickel column affinity chromatography. The purified products were identified by 12 alkyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Mouse polyclonal antiserum was prepared by immunizing mice with purified protein. 8 patients clinically diagnosed as tuberculosis pleural effusion (TB group) and 8 adenocarcinomas patients (CA group) were enrolled and their pleural effusion and plasma were collected. 8 healthy people (HC group) were enrolled as the control group and their plasma were collected. An indirect ELISA was used to detect the level of specific antibodies recognizing MPT83 protein in the samples. Results Mtb MPT83 protein was successfully expressed and purified. The serum titer of MPT83 mouse polyclonal antibody was as high as 1∶1 280 000. The plasma levels of MPT83 antigen specific antibodies in TB group were significantly higher than those in HC group (P<0.05), while the plasma levels of MPT83 antigen specific antibodies in CA group were not significantly different from those in HC group (P>0.05). Compared with the HC group, there was no significant difference in pleural fluid in both the TB and CA groups (P>0.05). The ROC curve was used to analyze the OD values of plasma in TB group and HC group, and the area under the curve was greater than 0.7, showing high diagnostic efficacy. Conclusion　MPT83 protein has high antigen specificity and immunogenicity, which has great application value in the immunological diagnosis of tuberculosis.

Objective To explore the relationship and clinical significance of Gli1 expression in peripheral blood and cancer tissues of gastric.Method 95 gastric cancer patients were recruited as surgery group,and 40 recurrent gastric cancer patients were recruited as recurrent group.50 normal subjects were recruited as control group.Paraffin specimens of gastric cancer tissues and gastric mucosa adjacent to cancer were collected to test expression of Gli1 protein.ELISA was applied to test Gli1 in peripheral blood.Results Levels of Gli1 in peripheral blood of preoperative surgery group were higher than in control group [(9.21 ± 1.37) ng/ml vs.(0.79 ±0.18) ng/ml,t =43.193,P ＜0.001].Post-up Gli1 decreased significantly to the level of control group (t =0.722,P =0.472).Levels of Gli1 in peripheral blood of recurrent group were higher than that in preoperativesurgery group (t =14.575,P ＜ 0.001).Gli1 protein in cancer tissues was higher than that in adjacent gastric mucosa (t =33.471,P ＜0.001),and Gli1 protein in cancer tissues of surgery group was higher than that in cancer tissues of recurrent group (t =8.808,P ＜0.001).Conclusion In postoperative gastric cancer patients increased Gli1 level in peripheral blood and cancer tissues predicts recurrence.

Objective To construct luciferase reporter plasmids of truncated fragments of different lengths of human guanylate binding protein 5(GBP5)gene promoter and analyze the transcriptional activity of each fragment to determine the core regulatory region.Methods GBP5promoter sequence was amplified by PCR,truncated into five fragments of different lengths and connected to pGL3-basic plasmid.The constructed recombinant plasmids pGL3-GBP5-11/21/31/41/51were transfected into 293FT cells and detected for luciferase activity.The binding sites of transcription factors in GBP5promoter region were predicted by JASPAR software,and Yin-Yang transcription factor 1(YY1)targeting the core regulatory region was selected and verified for the transcriptional regulatory activity.The CDS sequence of YY1 was amplified by PCR to construct the overexpression plasmid pIRES2-EGFP-YY1,which was then co-transfected to 293FT cells with plasmids pGL3-GBP5-21(-1 623 ～ +47 bp)and internal reference plasmid pRL-CMV,and detected for luciferase activity to analyze the regulation of transcription factor YY1 on GBP5 promoter activity.Results Colony PCR and double enzyme digestion identification proved that the plasmid of human GBP5 promoter reporter gene was correctly constructed;JASPAR software predicted that there were multiple transcription factor binding sites such as STAT1,YY1 and Foxp3 in GBP5promoter region.Double luciferase activity assay showed that pGL3-GBP5-21(-1 623 ～ +47 bp)showed the highest promoter activity,while the promoter activity of pGL3-GBP5-41(-520 ～ +47 bp)decreased significantly,suggesting that the core region of GBP5 promoter was located at upstream-1 623 ～-520 bp of 5 'UTR;Overexpression of YY1 significantly activated the GBP5 promoter activity and regulated the expression of GBP5.Conclusion The core regulatory region of human GBP5 promoter was located in upstream-1 623 ～-520 bp of the 5 'UTR,with a binding site of transcription factor YY1 existing in this region.Meanwhile,overexpression of YY1 significantly effected the activity of GBP5 promoter.

Objective To retrieve,evaluate and summarize the best evidence from home and abroad on the management of enteral nutrition complicating gastric retention in critically ill patients,and to provide a reference basis for clinical nursing staff to manage enteral nutrition complicating gastric retention in critically ill patients.Methods We searched websites and databases,including UpToDate,BMJ Best Practice,National Institute for Health and Clinical Excellence,Guidelines International Network,Registered Nurses Association of Ontario,Scottish Intercollegiate Guidelines Network,JBI,World Health Organization,Yimaitong guide network,Cochrane Library,PubMed,Embase,Web of Science,Sinomed,CNKI,Nutrition Society website,CINAHL(EBSCO),Wanfang Database,and collected relevant clinical decisions,guidelines,evidence summaries,expert consensuses and systematic reviews.2 researchers independently assessed methodological quality of included papers and extracted data.Results A total of 18 publications were included,including 3 guidelines,4 expert consensuses,4 systematic reviews,5 randomized controlled trials,1 quasi-experimental study,and 1 best evidence summary.18 pieces of the best evidence were summarized in 5 areas,including identification of gastric retention,management of enteral nutrition,management of monitoring,management of threshold determination,and management of treatment.Conclusion This study provides evidence-based evidence for nursing staff to manage enteral nutrition in critically ill patients with concomitant gastric retention,which can help reduce the incidence of gastric retention in critically ill patients.