This study aims to establish an antibody detection method for porcine deltacoronavirus (PDCoV). The recombinant proteins PDCoV-N1 and PDCoV-N2 were expressed via the prokaryotic plasmid pColdII harboring the N gene sequence of the PDCoV strain CH/XJYN/2016. The reactivity and specificity of PDCoV-N1 and PDCoV-N2 with anti-PEDV sera were analyzed after the recombinant proteins were analyzed by SDS-PAGE and purified by the Ni-NTA Superflow Cartridge. Meanwhile, Western blotting and indirect immunofluorescence assay were carried out separately to validate the recombinant proteins PDCoV-N1 and PDCoV-N2. Finally, we established an indirect ELISA method based on the recombinant protein PDCoV-N2 after optimizing the conditions and tested the sensitivity, specificity, and reproducibility of the method. Then, the established method was employed to examine 102 clinical serum samples. The recombinant protein PDCoV-N2 showed low cross-reactivity with anti-PEDV sera. The optimal conditions of the indirect ELISA method based on PDCoV-N2 were as follows: the antigen coating concentration of 1.25 μg/mL and coating at 37 ℃ for 1 h; blocking by BSA overnight at 4 ℃; serum sample dilution at 1:50 and incubation at 37 ℃ for 1 h; secondary antibody dilution at 1:80 000 and incubation at 37 ℃ for 1 h; color development with TMB chromogenic solution at 37 ℃ for 10 min. The S/P value ≥ 0.45, ≤0.38, and between 0.45 and 0.38 indicated that the test sample was positive, negative, and suspicious, respectively. The testing results of the antisera against porcine epidemic diarrhea virus (PEDV), porcine circovirus 2 (PCV2), transmissible gastroenteritis virus (TGEV), foot-and-mouth disease virus (FMDV), and African swine fever virus (ASFV) showed that the S/P values were all less than 0.38. The testing results of the 800-fold diluted anti-PDCoV sera were still positive. The results of the inter- and intra-batch tests showed that the coefficients of variation of this method were less than 10%. Clinical serum sample test results showed the coincidence rate between this method and neutralization test was 94.12%. In this study, an ELISA method for the detection of anti-PDCoV antibodies was successfully established based on the truncated N protein of PDCoV. This method is sensitive, specific, stable, and reproducible, serving as a new method for the clinical diagnosis of PDCoV.
Animals ; Enzyme-Linked Immunosorbent Assay/methods* ; Swine ; Antibodies, Viral/blood* ; Recombinant Proteins/genetics* ; Deltacoronavirus/isolation & purification* ; Coronavirus Infections/virology* ; Swine Diseases/diagnosis* ; Coronavirus Nucleocapsid Proteins ; Sensitivity and Specificity

Objective To explorethe long-term prognosis of the triangular resectionvalvuloplasty for posterior mitral valve prolapse and provide a reference for the clinical treatment.Methods We retrospectively analyzed the clinical data of 61 pa-tients with posterior mitral valve prolapse underwent the mitral valvuloplasty in our hospital from Match 2008 to December 2016.30 patients who underwent triangular resection for posterior mitral valve prolapse.31 patients who underwentstandard quadrangular resection.The cardiac functional parameters and condition of mitral regurgitation were compared among the period of preoperation,discharging and follow-up.Results The mean follow-up period was 42 ±34(3-106)months.left atrium and left ventricular diameter were decreased(P<0.05).Inresrarch group,there was 1 patient who have moderate regurgitation,ar-rhythmia is one.In control group,there was arrhythmia in 1 cases,moderate regurgitation in 2 cases and death in 1 cases.Rate of freedom from reregurgitation was(97 ±6)% in research group and was(97 ±4)% in control group.There was no SAM in two groups.There were no significant differences of survival or risk of reoperation.There was a statistic difference of NYHA classification of cardiac function in the phase of follow-up than that of discharging(P<0.05).Conclusion Triangular resec-tion of a prolapsed posterior mitral leaflet is a simplified,reliable, reproducible, and durable procedure,like quadrangular re-section of a prolapsed posterior leaflet.

Objective:To investigate the effect of up‐regulating miR‐519d on proliferation and apoptosis of laryngeal squamous cell carcinoma .Methods:The expression of signal transducer and activator of transcription 3 (STAT3) and miR‐519d in human laryngeal carcinoma epithelium cell line Hep‐2 and human bronchial epithelium cell (HBE ) were detected by real‐time fluorescent quantitative polymerase chain reaction (RT‐PCR) and Western blotting .After up‐regulating miR‐519d expression in Hep‐2 cell line by plasmid transfection technique ,the proliferation and apoptosis of transfected cells and control cells was detected by proliferation and apoptosis assays .Results:STAT3 mRNA expression in Hep‐2 cells was significantly higher than that in HBE cells (P<0 .05) .However ,miR‐519d expression in Hep‐2 cells was significantly lower than that in HBE cells (P<0 .05) .The STAT3 mRNA expression in the transfected Hep‐2 cells decreased significantly .During 0 and 7 day of post‐transfection culture ,proliferation rate of transfected Hep‐2 cells was significantly lower than that of untransfected cells (P<0 .05) .After 24 h culture ,the apoptosis rate of miR‐519d transfected cells was (2 .8 ± 0 .15)% ,while that of control cells was (0 .92 ± 0 .09)% (P=0 .000 4) .After 72 h culture ,the apoptosis rate of miR‐519d transfected cells was (23 .06 ± 3 .52)% , while that of control cells was (23 .26 ± 2 .56)% (P=0 .965 5) .Conclusions:STAT3 shows high expression in Hep‐2 cell , while the related miR‐519d shows low expression .By up‐regulating miR‐519d ,STAT3 expression could be suppressed ,so as to suppress the proliferation of tumor cells and promote the apoptosis of tumor cells .

Objective To evaluate the clinic effect of leaflet enlargement with autologous pericardium in repairing mitral valve disease and to describe the technique and discuss its indications. Methods Between July 2004 and June 2008, 45 pa-tients with isolated mitral valve disease, included stenosis in 10 and regurgitation in 35. The causes were congenital heart dis-ease in 8, rheumatic in 21, degenerative in 7 and endecarditis in 9. The procedures were: posterior leaflet enlargement with autologuus pericardium in 14, anterior leaflet enlargement in 8, both anterior and posterior leaflet enlargement in 23. In addi-tion, eye to eye technique was in 12, artificial chordal in 12, chordal transfer in 6, papillary muscle vepesitioning in 4. Mitral anuuloplasty was performed in all cases. Before and after surgery, cardiac function parameters were compared. Results No operative deaths occurred. One case wastransfered to mitral valve replacement due to regurgation, lntraoperative transesophageal echocardiography showed no mitral regurgitation in 38 and small regurgitation in 6 cases. The mean mitral valve effective orifice area(MVEOA) was (2.8±0.6) cm~2, with a mean gradient pressure of (6.21±1.34) mm Hg after operation. The mean followed up was ( 18.0±2.1 ) months. Echocardiography study showed that no mitral regurgitation in 35 cases, slight regurgi-tation in 9, mean mitral effective orifice area was (2.5±0. 8 ) cm~2, mean gradient pressure of (7.21±0. 45 ) mm Hg, no one need reoperation. Postoperative cardiac functions were significantly improved: the average left ventricular end-diastolic diameter (LVEDD) was (48±7) mm [ preoperative (56±6) nun, P <0.05], ejection fraction (EF) was 0.51～0.24( preoperative 0.45± 0.23, P < 0.05 ), the average left atrium diameter ( LA ) was ( 50±11 ) mm [ preoperative ( 62±23 ) mm, P <0. 05 ]. The function of mitral valves was well performed. Conclusion Leaflet enlargement with autologous pericardium com-bined with mitral annuloplasty was effective in repairing of diseased mitral valve. The advantages of the procedure including simplicity, good compatibility, avoiding foreign body and no need for anticoagulation.

Objective To analyze the adverse drug reactions(ADRs)of edaravone in treatment of patients with acute cerebral infarction(ACI)by reviewing Chinese medical literature and to evaluate safety of edaravone treatment in ACI.Methods Publications from Pubmed and Chinese Biomedical Literature Datababe(CBMdisc)were reviewed and the ADR of edaravone was analyzed among the published 8645 cases.Literatures about randomed-control clinical trails(RCTs)on security of edaravone for treating ACI Was analyzed by meta-analysis.Results Abnormal hepatic function,especially mild elevation of aminotransferase,renal dysfunction and skin rash induced by edaravone were tIle most common ADRs.Among 8645 patients,ADRs were reported in 283(3.27 % ).The meta-analysis in RCTs showed that between the group treated with a combination of edaravone and routine and the group treated with routine treatment only,there was no significant difference in the occurrence rate of ADRs(OR=1.18,95 % CI0.70-2.00,P=0.536),elevation of aminotransferase(OR=1.23,95 % CI 0.57-2.68,P=0.595)or renal dysfunction including albuminutia,increased level of serum ereatinine and nitrogen(OR=1.65.95 % CI0.57- 4.79.P=0.353).Conclusion Edaravone has a low ADRs OCCurrence rate and iS safely used in cerebral infarction treatment.

Objective To establish and optimize the technology and method of producing large quantity and high-paclitaxe yield callus of 〖WTBX〗Taxus chinensis var. mairei. Methods Wild maternal tree grown in Lingchuan County of Shanxi Province and cultivated tree grown in Xi′an were used as explant source. And the optimum maternal tree for explant cutting, optimum explant type, basic medium, composition and concentration of growth regulators in medium and so on, which were factors of affecting on callus induction, growth and paclitaxe yield, were examined in a series order. Results The juvenile stem segments were the optimum explants because of their earlier and higher rate callus induction than that of other explants. Medium Y5: MS+2,4-D 4.0 mg/L+KIN 1.0 mg/L or medium B5 Ⅲ: B5+2,4-D 3.0 mg/L+KIN 0.1 mg/L+Phe 0.1 mol/L was confirmed optimum callus induction medium in which callus induction rate had reached to 100%. In callus subculture medium, lower concentration of 2,4-D (0.5—3 mg/L) always increased callus growth, but higher concentration of 2,4-D (8 mg/L) reduced callus growth. When 2,4-D concentration was suitable, callus grown on B5 medium displayed lighter browning and faster tissue growth than that on MS medium. Further more, HPLC analysis confirmed that the paclitaxel yield in callus grown on medium MSⅢ was highest and had reached 0.004% of callus dry weight. In a general condition, the level of paclitaxel in calli derived from juvenile stems of wild maternal tree was higher than that in calli initiated from cultivated maternal tree's juvenile stems. Conclusion The optimization sequence of obtaining a large quantity and high-paclitaxe yield callus of T. chinensis var. mairei are dividing juvenile stem segments from wild maternal tree in May and culturing on medium Y5 or B5 Ⅲ for callus induction. After the calli having been subcultured on the same medium for 8—10 generations, one or two generations are recultured on medium MSⅢ. Finally, the calli with more paclitaxel are obtained by extracting paclitaxel out of it.