Objective:To prepare GST-Orai2 fusion protein and to prepare polyclonal antibody against Orai2 by immunizing rabbits.To further investigate the function of Orai2,a transmembrane protein,this antibody was used to identify the Orai2 conditional gene knockout mice.Methods:ORF of Orai2 was amplified by PCR and subcloned into pGEX-6p-1 vector.After transforming BL21 (DE3) competent cells,we succeeded in inducing the expression of GST-Orai2 fusion protein using IPTG.Then the GST-Orai2 protein was purified by immobilized Glutathione affinity chromatography and identified by SDS-PAGE.A New England rabbit was immunized with the prepared fusion protein in Freund's adjuvant to prepare specific antibody.Finally,the prepared antibody was identified by Western blot by checking its titer and specificity.Furthermore,we took use of the prepared Orai2 antibody to identify Orai2 conditional gene knockout mice,with comparing to the wildtype ones in the same cage.Results:The purity of purified GST-Orai2 reached to 90% and the concentration was 0.35 mg/ml by BCA kit.We could detect Orai2 protein even in dilution of 1:10,000.Also,the prepared polyclonal antibody agianst Orai2 could detect both overexpressed and endogenous Orai2 protein in mouse-brain,without crossing reaction with Orai1.As well,we found that the Orai2 protein expression was of obvious reduction in Orai2 conditional gene knockout mice,compared with the wildtype ones in the same cage.Conclusion:We successfully obtain the purified GST-Orai2 fusion protein and prepare specific and highly sensitive polyclonal antibody against Orai2.The antibody can be used to detect overexpressed and endogenous Orai2 protein inmouse-brain specifically,and to identify Orai2 conditional gene knockout mice,without any crossing reaction with Orai1.Our work contributes a lot to the future investigation of functions of Orai2.

AIM: To investigate the effect of 3-methyladenine (3-MA) combined with trichostatin A (TSA) on triple-negative breast cancer cells and the mechanism involved.METHODS: The viability of MDA-MB-231 cells was detected by MTT assay, the migration ability was determined by scratch assay, and the expression of autophagy-related proteins was detected by Western blot.RESULTS: TSA significantly inhibited the viability of MDA-MB-231 cells in a time-and dose-dependent manner.The results of scratch assay showed that TSA inhibited cell migration ability.Western blot data indicated that TSA resulted in a moderate increase in LC3-Ⅱ expression.Moreover, 3-MA inhibited cell autophagy induced by TSA, and combination of 3-MA and TSA further inhibited the viability of MDA-MB-231 cells.CONCLUSION: Combination of 3-MA and TSA may effectively inhibit the growth of triple-negative breast cancer cells.

Objective: To assess the effectiveness of interventions on patient trust Methods: A system review was conducted through searching 11 relevant databases, such as Cochrane Library, EMBASE, PsycINFO, CINAHL, PubMed, Web of Science, SinoMed, CNKI, WANFANG. The randomized controlled trials (RCTs) were screened from 1539 original studies according to inclusion and exclusion criteria Meta-analysis was used to evaluate and interpret the interventions. Results: Ten RCTs were included in the review. It was concluded from meta-analysis that no significant difference could be found between controlled and intervention groups by increasing doctors'abilities(SMD = -0.05, 95％ CI: -0.22 -0.12); while the other two interventions, doctor's information disclosure as well as patient education and nursing intervention, significantly enhanced patient trust (SMD = 0.13, 95％ CI: 0.06 - 0.21; SMD = 0.60, 95％ CI: 0.29 - 0.90). Conclusion: Doctor's information disclosure, patient education and nursing intervention are effective in increasing patients'trust, while intervention of doctor's ability promotion seems not so remarkable.

Objective To summarize the methods and experiences of one-stage repair of aortic coarctation.Methods During January 2009 and September 2016,31 patients underwent surgery for coarctation of aorta (COA) in our department.Of them,fifteen patients were accompanied by patent ductus arteriosus (PDA),five associated with ventricular septal defect (VSD),five with VSD and PDA,and six patients were single COA.All patients accepted one-stage repair and follow-up.The surgical strategies and follow-up results were retrospectively reviewed.Results Excision of coarctation and anastomoses was performed in 24 patients,a bypass with artificial graft was performed in 3 patients,longitudinally section transverse suture was performed in 2 patients,and aortoplasty with patch graft was used in 2 patients.Accompanied intracardiac malformations were repaired simutaneously.One patient died of low cardiac output syndrome 1 d after operation.Other patients recovered smoothly.Thirty patients were followed up from 6 months to 7 years.The anastomotic systolic pressure difference disappeared in 21 patients.Eight patients still suffered from systolic pressure difference (5-20 mmHg),but had no need of reoperation.One patient suffered from anastomotic recoarctation 2 years and 3 months after operation and the pressure difference was 48 mmHg.After exerting balloon dilatation,the pressure difference reduced to 17 mmHg.Conclusions Surgery is an effective method of COA treatment.The surgical strategies should be selected according to the characters of coarctation lesions,intracardiac malformations,and the age of patient.

Objective:To explore the diagnostic value of Caprini score for acute pulmonary embolism(APE).Methods:Totally 2 764 patients with suspected APE admitted to Yuncheng Central Hospital were enrolled from January 2012 to January 2019, and 312 patients were diagnosed APE and assigned to APE group finally.Among the patients without APE, 312 patients(control group) were matched with the patients in APE group according to age, gender, weight, blood pressure, surgical history, etc.The general clinical data of the two groups were collected, and the Caprini score of each patient was recorded.The differences between the two groups in clinical data and Caprini score were compared.The area under curves(AUC) of receiver operating characteristic(ROC) was calculated to predict the diagnostic efficacy of Caprini scale for patients with APE.Results:The Caprini score of the APE group was significantly higher than that of the control group[(5.41±2.47)points vs.(2.16±1.28)points, t=1.180, P=0.004]. The Caprini score had a favorable diagnostic efficacy for patients with suspected APE(AUC=0.915, 95% CI: 0.878-0.995, P<0.001), and when the 3.5 cutoff value of Caprini score was determined, the specificity and sensitivity were 87.76% and 95.24%, respectively, with 7.778 of positive likelihood ratio, 0.054 of negative likelihood ratio, and 0.83 of Youden index. Conclusion:Caprini score has strong diagnostic efficacy in patients with APE.