In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

Objective To improve diagnosis on esophageal tuberculosis by the endoscopic ultrasonography (EUS).Methods A retrospective analysis of 9 cases of esophageal tuberculosis patients diagnosed by EUS or EUS guided ifne needle aspiration (EUS-FNA) who were treated from 2011 to 2016 and literature reviewed.Results The main presenting symptoms were dysphagia (6 cases), burning pain after sternum (3 cases), low-grade fever and night sweat (2 cases), cough and hemoptysis (1 case). The middle esophagus was involved in 7 cases, upper and lower third of the esophagus were affected in 1 cases. The endoscopic appearances included ulcer (1 case), mass lesion (8 cases). The diagnoses were conifrmed by biopsies with pathology in 5 cases. The layers of the esophageal wall were unclear or disappeared in some cases, and in some patients low echo occupying lesions inside or outside the esophageal wall were showed by EUS. The internal echo of the lesions was heterogeneous, and strong echo spots could be observed. In most cases, enlarged mediastinal lymph nodes were observed. EUS-FNA was used to take biopsy specimens in 2 cases: biopsy specimens suggested tuberculosis; and anti-TB treatment has good effect as well.Conclusion Dysphagia is the major clinical symptom of esophageal tuberculosis, always inlfuences the middle part of the esophagus. The commonest abnormalities identiifed by endoscopy are ulceration and mass lesion. The diagnostic rate of the disease can be improved by endoscopic examination with biopsies, EUS and EUSFNA data.

Objective To investigate the clinical efficacy of conventional treatment combined with flupentixol and melitracen in patients with reflux esophagitis (RE).Methods From June 2012 to March 2015, a total of 182 patients were selected as study subjects from newly diagnosed RE patients.The anxiety and depression scores were evaluated according to Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD).And then patients were divided into HAMA and HAMD negative conventional treatment group and combined treatment group, HAMA and HAMD positive conventional treatment group and combined treatment group.Rabeprazole and mosapride were administrated in conventional treatment group.For patients in combined treatment group, on the base of conventional treatment flupentixol and melitracen were added.The treatment course was eight weeks.The degree of anxiety and depression, RE symptoms and mucosal healing under gastroscope were evaluated before and after treatment.Adverse drug reaction was observed.Chi square test or t test was performed for statistical analysis.Results Eight weeks after treatment, the scores of HAMA and HAMD in HAMA and HAMD positive combined treatment group were 7.930 ±3.832 and 9.630 ± 3.650, which were both lower than those of conventional treatment group (11.660 ± 4.108 and 12.170 ± 4.459), and the differences were statistically significant (t=3.683 and 2.233;both P＜0.05).The symptom scores of heartburn, regurgitation, chest pain of HAMA and HAMD positive combined treatment group were 0.700±0.591,0.780± 0.629 and 0.720±0.621, respectively, which were lower than those of conventional treatment group (1.280 ± 0.502, 1.370 ± 0.610 and 1.040 ± 0.842), and the differences were statistically significant (t =5.133, 4.413 and 2.114, all P＜0.05).There were no statistical significance in symptoms scores between HAMA and HAMD negative combined treatment group and conventional treatment group (all P＞0.05).After treatment, the mucosal healing rate of HAMA and HAMD positive combined treatment group was 91.3％ (42/46), which was higher than that of conventional treatment group (71.7 ％, 33/46), and the difference was statistically significant (x2 =5.845, P =0.016).The incidence of adverse events of HAMA and HAMD negative combined treatment group was 4.8％ (2/42), and that of HAMA and HAMD positive combined treatment group was 2.2％(1/46).Conclusions The conventional treatment combined with flupentixol and melitracen in RE patients accompanied with anxiety and depression was remarkable and safe.RE patients without obvious anxiety or depression, preventive use of antianxiety and antidepressant medicine can not improve the efficacy.

Objective To study whether citrate lock is superior to heparin lock in the prevention of catheter related infections,bleeding complications and catheter malfunctions among hemodialysis patients with tunneled catheters.Methods By searching in Pubmed,the Cochrane Library,EMBASE,Ovid,WanFang,VIP,CNKI and CBM databases as well as related journals,qualified randomized controlled trials were included in a Meta-analysis using Revman 5.0 and STATA 10.0 software.The endpoints included catheter related infection,bleeding complication,thrombolytic treatment,catheter removal for malfunction,catheter thrombosis and all-cause death.Results Fifteen randomized controlled trials were included with 1621 patients involved.Eight studies compared citrate alone with heparin lock,while 7 trials focused on citrate in combination with other antimicrobials.Pooled analysis demonstrated that incidence of catheter related infections in patients receiving citrate lock decreased by 47％ compared with those on heparin (RR=0.53,95％CI 0.36-0.77,P ＜ 0.01).Subgroup analysis by types of citrate lock indicated that all combined lock solutions of citrate and other antimicrobials (citrate + gentamicin,citrate + taurolidine,citrate + methylene blue +methylparaben + propylparaben) were superior to heparin lock in preventing catheter-related infections (P =0.01,0.04,0.01,respectively); citrate alone seemed to reduce catheter-related infection risk (RR =0.68),but no statistically significant difference was observed (95％CI 0.38-1.21,P=0.19).There were fewer patients with bleeding complications in citrate group (RR =0.53,95％ CI 0.34-0.S4,P ＜ 0.01),while citrate showed no advantage over heparin lock in terms of thrombolytic treatment (P =0.93),catheter removal for malfunction (P =0.35),catheter thrombosis (P =0.64) and all-cause death (P =0.35).Conclusions For hemodialysis patients with tunneled catheters,combined lock solutions of citrate and other antimicrobials,rather than citrate alone,are superior to heparin in preventing catheter related infections.Citrate locks are associated with less bleeding complications,and are comparable to heparin in the maintenance of catheter patency.