In order to establish a serological method for the detection of bovine respiratory syncytial virus,the prokaryotic expression of four proteins of BRSV,G,F,P,and M was carried out,and the most suitable coating antigen was screened to establish an indirect ELISA detection method.The results showed that the four recombinant proteins of BRSV,rG,rF,rP and rM were successfully expressed.The results of checkerboard screening showed that the P/N value of rG protein was the largest,which was determined to be the best coating antigen established by indirect ELISA meth-od.The optimal reaction conditions for indirect ELISA were as follows:the mass concentration of rG protein coating was 1 mg/L,37℃ for 2 h;3％BSA 37℃ block for 1 h;Serum was diluted 1∶50 and incubated at 37℃ for 1h;Secondary antibody 1∶5 000 dilution,37℃ for 30min;The color development conditions of the substrate were 37℃ for 15 min;Thirty negative sera were selected,and the cut-off value was determined to be 0.63 by the established indirect ELISA method.The re-sults of the specificity test showed that the indirect ELISA method established in this test only recognized BRSV-positive serum,and did not react with IBRV,BCoV,and BPIV3-positive serum.The results of repeatability test showed that the method had good repeatability,and the coefficient of variation within and between batches was less than 10％.The results of the sensitivity test showed that the BRSV-positive serum was still positive when diluted to 1∶8 192.The indirect ELISA method established in this experiment was used to detect 100 clinical serum samples at the same time,and the total coincidence rate of the two reached 90.48％,the positive coincidence rate was 93.42％,and the negative coincidence rate was 82.75％.The indirect ELISA established in this test can be used for the detection of bovine respiratory syncytial virus in clinical practice.

In order to establish a serological method for the detection of bovine respiratory syncytial virus,the prokaryotic expression of four proteins of BRSV,G,F,P,and M was carried out,and the most suitable coating antigen was screened to establish an indirect ELISA detection method.The results showed that the four recombinant proteins of BRSV,rG,rF,rP and rM were successfully expressed.The results of checkerboard screening showed that the P/N value of rG protein was the largest,which was determined to be the best coating antigen established by indirect ELISA meth-od.The optimal reaction conditions for indirect ELISA were as follows:the mass concentration of rG protein coating was 1 mg/L,37℃ for 2 h;3％BSA 37℃ block for 1 h;Serum was diluted 1∶50 and incubated at 37℃ for 1h;Secondary antibody 1∶5 000 dilution,37℃ for 30min;The color development conditions of the substrate were 37℃ for 15 min;Thirty negative sera were selected,and the cut-off value was determined to be 0.63 by the established indirect ELISA method.The re-sults of the specificity test showed that the indirect ELISA method established in this test only recognized BRSV-positive serum,and did not react with IBRV,BCoV,and BPIV3-positive serum.The results of repeatability test showed that the method had good repeatability,and the coefficient of variation within and between batches was less than 10％.The results of the sensitivity test showed that the BRSV-positive serum was still positive when diluted to 1∶8 192.The indirect ELISA method established in this experiment was used to detect 100 clinical serum samples at the same time,and the total coincidence rate of the two reached 90.48％,the positive coincidence rate was 93.42％,and the negative coincidence rate was 82.75％.The indirect ELISA established in this test can be used for the detection of bovine respiratory syncytial virus in clinical practice.

Objective:To provide reference for the pharmaceutical care for aged acute cerebral infarction with various complica-tions. Methods:Clinical pharmacists participated in the treatment for an elder patient with acute cerebral infarction complicated with gastrointestinal bleeding, liver dysfunction and pulmonary infection through providing drug therapy information, analyzing the effect and safety of drug therapy and adjusting the pharmaceutical care plan according to the drug efficacy in time. Results:Taking use of phar-maceutical knowledge, clinical pharmacists provided whole-course pharmaceutical care for the treatment of the patient and coordinated with doctors to make the drug therapy, which guaranteed the effectiveness and safety of therapy. Conclusion:The old are with high in-cidence of cerebral infarction, and complications are common. Due to their physiological dysfunction and the other reasons, high safety risk exists in their drug therapy. Clinical pharmacists can use their professional knowledge to provide whole-course pharmaceutical care for the patients in order to improve the therapeutic effect and promote the rational drug use.

OBJECTIVE: To observe the effects of Xuefu Zhuyu Capsule (XFZYC), a compound traditional Chinese herbal medicine, on endothelin-1 (ET-1) release in myocardium and vascular endothelium and nitric oxide (NO)/nitric oxide synthase (NOS) system of swines after acute myocardial infarction (AMI) and reperfusion, and to explore the action mechanisms of XFZYC in improving the endothelium function. METHODS: Forty-five Yorkshire swines were randomized into 3 groups: sham-operated group, untreated group and XFZYC-treated group. A Yorkshire swine model of reperfusion in AMI was established by ligation of left anterior descending coronary artery for 90 min followed by 2 h relaxation. The content of serum ET-1 and NO was measured by radioimmunoassay before and after AMI and after reperfusion, respectively. Twenty-four hours after operation, all Yorkshire swines underwent diagnostic coronary angiography to delineate coronary arteries. The expressions of ET-1 and endothelial nitric oxide synthase (eNOS) in myocardial tissue of ischemic area were quantified with Western blotting. Microvessel density of the implanting sites was assessed by using HE staining. RESULTS: Compared with the untreated group, the levels of serum ET-1 after AMI and reperfusion were significantly decreased in XFZYC-treated group (P<0.01), while the NO levels after AMI and reperfusion in XFZYC-treated group were significantly increased (P<0.01). There was no significant difference in diagnostic coronary angiography between XFZYC-treated group and untreated group (P=0.253). Western blotting showed that the level of ET-1 in ischemic area in XFZYC-treated group was lower than that in the untreated group (P<0.01), while the eNOS protein expression in XFZYC-treated group was higher than that in the untreated group (P<0.01). The results of HE staining and microvessel density analysis of the implanting sites all showed that the degree of telangiectasis was reduced, the cardiac muscle damage was improved, and the density of capillaries was increased obviously in XFZYC-treated group as compared with the untreated group. CONCLUSION: The endothelium injury may be one of the important mechanisms for no-reflow phenomenon. XFZYC may reduce the no-reflow by protecting endothelium cells.