In order to develop a double antibody sandwich assay (DAS-ELISA) for detecting bovine respiratory syncytial virus (BRSV),New Zealand white rabbits and BALB/c mice were immunized with the purified G protein as an antigen to prepare anti-G protein polyclonal and monoclonal antibodies.The antibody concentration and reaction conditions of DAS-ELISA were optimized by square titration,and its sensitivity,specificity,and coincidence rate were validated.Five hybridoma were stably secreting Mab which subclass belonged to IgG1κ.Western blot and IFA test showed that PcAb and Mab could react specifically with G protein and BRSV.The PcAb and Mab as the capture antibody and detection antibody respectively,and their optimal working concentrations were determined to be 2.5 μg/mL and 10μg/mL,the critical value 0.22 and the detection limit of 1.43 μg/mL,batch,inter-assay coefficient of variation less than 10 ％.The DAS-ELISA had no cross-reaction with several pathogens which often caused bovine respiratory disease.When 45 nasal swabs of clinical samples were simultaneously detected by the DAS-ELISA and RT-PCR,the sensitivity,specificity and coincidence rate were 92.0 ％,100 ％,95.6 ％,respectively.It' s indicated that the established DAS-ELISA detection method can be used to detect a large number of clinical samples.It was the foundation of monitoring and quick diagnosis for BRSV.

This study was designed to investigate the effects of wheat grain moxibustion on the immune function of mice immunosuppressed by cyclophosphamide,and to delve into the potential mechanisms by analyzing factors related to NF-κB signaling pathway.Sixty SPF-grade ICR mice were randomly allocated into four groups:blank group,model group,traditional Chinese medicine group and wheat grain moxibustion group,with 15 mice in each group.Immunosuppression was induced in mice of the model,traditional Chinese medicine and wheat grain moxibustion groups by intraperitoneal injection of cyclophosphamide at 80 mg/kg daily for three days,while mice of the blank group received an equivalent volume of saline.The traditional Chinese medicine group was treated with oral administration of Zhēn Qí Fú Zhèng granules,and the wheat grain moxibustion group was treated with wheat grain moxibustion at"Da Zhui,"bilateral"Zu San Li,"and bilateral"San Yin Jiao"once a day for seven consecutive days.After corresponding treatment,spleens were harvested from the mice of different groups for pathological examination,and enzyme-linked immunosorbent assay(ELISA)was utilized to measure the serum levels of IL-2,IL-4,TNF-α and IFN-γ.Additionally,Western blot analysis was conducted to assess the protein expression levels of IκBα and P-IκBα in spleen tissue.Compared with the blank group,mice in the model group exhibited poorer general conditions,significantly reduced white blood cell(WBC)count(P＜0.01),extensive damage in spleen tissue,significant decreases in serum IL-4 and IL-2 concentrations,increases in TNF-α and IFN-γ levels(P＜0.01),lower level of P-IκBα protein in spleen tissue(P＜0.01),and higher level of IκBα protein(P＜0.01).These changes mentioned above were reversed in mice of both the wheat grain moxibustion group and the traditional Chinese medicine group(P＜0.01).Compared to the traditional Chinese medicine group,on the last day of treatment,the wheat grain moxibustion group showed slightly higher WBC counts(P＜0.05),better tissue repair,and lower levels of IFN-γ(P＜0.01).In conclusion,wheat grain moxibustion can affect the activity of NF-κB signaling pathway by regulating the relative expression of IκBα and P-IκBα,and then regulate the immune response of the body,thus play a role in alleviating the immunosuppression caused by cyclophosphamide.

Objective To evaluate the role of programmed cell death ligand-1 (PD-L1) in a mouse model of bone cancer pain.Methods Ninety-six male C3H/HeN mice (20-25 g,4-6 weeks old),which inoculated with osteolytic NCTC 2472 cells,were used to build the model of bone cancer pain.Part one:sixtyfour male C3H/HeJ mice were randomly divided into sham group (group Sham,n =32) and tumor group (group Tumor,n=32).Part two:Twenty-four male C3H/HeJ mice which were inoculated with osteolytic NCTC 2472 cells were randomly divided into group T (tumor,n=8),group PD-L1 (intrathecal injection with PLX3397,1 μg/5μl,n=8) and group NS (intrathecal injection with normal saline,n=8).Also,there were eight male C3H/HeJ mice in group S which were intra-femur inoculated with α-MEM.The pain behaviors of Sham group and Tumor group were observed and the expression of PD-L1 was detected before inoculation and on 4,7,10,14 and 21 days after inoculation,including paw withdrawal mechanical threshold (PWMT) and the number of spontaneous flinches (NSF).On 14 d after inoculation,the mice of group PD-L1 and group NS were intrathecal injected with drugs respectively.Pain behaviors were observed before injection and 2,4,6,24h after injection.Results Compared with group Sham,PWMT was significantly decreased and NSF was increased on 7～ 21 d after inoculation in group Tumor (P＜0.05).Compared with baseline and group S (baseline (0.38±0.06),group Sham (0.35±0.08),(0.38±0.08),(0.36±0.07)),the expression of PDL1 was up-regulated on 10-21 d after inoculation in group Tumor ((0.77±0.06),(1.21±0.04),(1.18±0.06)) (P＜0.05).Compared with group NS,PWMT was significantly increased (group NS (0.25t0.12),(0.25±0.12),(0.31±0.12),group PD-L1 (1.43±0.49),(1.35±0.44),(0.95±0.26)),and NSF was decreased on 2-6 h after injection in group PD-L1 (group NS(11.74± 1.31),(13.78±0.0.91),(13.63±1.06),group P D-L1 (4.90± 0.82),(4.15± 0.71),(7.65±0.56)) (P＜0.05).Conclusion Expression of PD-L1 in spinal cord was up-regulated in the mouse model of bone cancer pain.Intrathecal injection of recombinant PD-L1 has an analgesic effect on mice with bone cancer.

As a chronic and lifelong disease， diabetes mellitus occurs across all age groups and gender groups. Since the disease requires lifelong treatment， it seriously affects the quality of life of patients. With the rising incidence on a global scale， diabetes mellitus has become a global problem that seriously affects public health. Moreover， the complications of this disease have aroused concern from the global medical research community， the World Health Organization， and the public. In the past， Western medicine was used in the clinical treatment of diabetes mellitus， which， however， had drug dependence， unsatisfactory efficacy， and side effects. Long-term oral administration of antidiabetics may cause liver and kidney function damage， hypoglycemia and other adverse symptoms. The treatment of diabetes mellitus has been faced with challenges such as limited efficacy and obvious side effects. Therefore， exploring more effective treatment means， especially tapping the potential of traditional Chinese medicine （TCM） in the treatment of diabetes mellitus， is a major issue to be solved. TCM has shown a great application value and a broad prospect in the treatment of diabetes mellitus because of multi-target regulation， a holistic view， synergistic effects， and high safety. TCM has a history of thousands of years in the prevention and treatment of diabetes mellitus， with rich experience accumulated and remarkable results achieved. Particularly， TCM demonstrates definite therapeutic effects on the complications. The application of TCM in the treatment of complications has been recognized and accepted by patients because of the definite therapeutic effect. In recent years， great progress has been achieved in the treatment of diabetes mellitus by the combination of Chinese and western medicine， which has made important contributions to the control of diabetes mellitus. This paper reviews the articles about the treatment of diabetes mellitus with TCM classic prescriptions， summarizes the treatment of clinical cases regarding the indications of these prescriptions， and provides an overview of the treatment mechanisms， aiming to offer fresh insights and strategies for the clinical diagnosis and treatment of diabetes mellitus.

Objective: To summarize the clinical experience of cardiopulmonary bypass(CPB) in acute type A aortic dissection received aortic arch reconstruction on age over 70 years patients. Methods: From April 2013 to December 2017, 35 elderly patients aged over 70 who were involved the aortic arch, brachiocephalic trunk, left common carotid artery and left subclavian artery and severe arch lesions, large false lumen and large rupture risk of acute A aortic dissection were reconstructed by triple-branched stent graft in emergency extracorporeal circulation. During the period of cardiopulmonary bypass, selective cerebral perfusion and discontinuous arrest under moderate hypothermia(25 degrees C) were used to reduce the time of cerebral ischemia and hypoxia, and to strengthen the management of the protection of important organs. Results: Cardiopulmonary bypass time was(144.85±32.98)minutes, and aortic cross clap time was(51.82±17.59)minutes, and selective cerebral perfusion time was(12.17±4.70)minutes, discontinuous arrest time was(4.50±3.54) minutes, the lower body arrest time was(16.6±7.49)minutes. All patients were smoothly weaned from cardiopulmonary bypass.35 patients resuscitated within 48 hours, of which 3 patients had transient mental disorders in the early stage and recovered before hospital discharge; 2 patients had cerebral infarction. Hospital mortality in this group was 17.1%(6/35). Conclusion Advanced age is not a contraindication to the reconstruction of the arch of the aortic dissection. Reasonable management of cardiopulmonary bypass can provide an important guarantee for the reconstruction of the elderly patients with acute type A aortic dissection.