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.

Objective To evaluate the outcomes of endoscopic sinus surgery (ESS) for children with chronic rhinosinusitis, who had no response to medication. Methods A total of 112 children (aged 12 to 17 years) with chronic rhinosinusitis caused by abnormal anatomy of the nose cavity were treated by ESS in our hospital. Septorhinoplasty was performed on the patients under an endoscope. Conchoplasty, adenoidectomy, or nasal polypectomy was performed simultaneously if necessary. Results The patients were followed up for 6-23 months (mean 13 months). The symptoms of rhinosinusitis disappeared in 110 patients (98.2%); whereas the other 2 patients showed no obvious improvement after the operation. Conclusions By using microinvasive ESS combined with conchoplasty, children with rhinosinusitis caused by abnormal anatomy that does not respond to medication could be cured.

Objectives To understand the epidemiological characteristics of Japanese Encephalitis in Shanghai and to provide evidence for preventing JE.Methods Epidemic characteristics,JEV antibody in healthy population and swine infection rate in Shanghai were analyzed by methods of field survey,serology and molecular biology.Results JE incident rate in Shanghai was 0.077/100,000 in 2006;and 0.129/100,000 in 2007.Antibody positive rate before JE epidemic fastigium was 60.39%;postive rate after epidemic was 85.44%.JE IgG positive rate was 26.92% in 3-month swine and 14.86% in swine for sale;JE Gene in mosquito was analyzed for type 1.Conclusion The JE prevalence rate is relatively low in Shanghai.JE antibody positive rate is high in Shanghai population.Swine as a media is infected by JE virus.The JE virus in mosquitoes belongs to genotype 1.

Poly (ADP-ribose) polymerase-1 (PARP-1) is a ribozyme that widely exists in the cukaryotic cells.It plays important roles in the maintenance of genomic stability and the regulation of gene transcription and other physiological processes.PARP-1 is overactivated after ischemic brain injury,and PARP-1 gene knockout mice or PARP-1 inhibitor can reduce brain injury in a variety of models of cerebral ischemia.Therefore,PARP-1 plays important roles in the pathophysiologic processes of ischemic brain injury.The investigation of the effect of PARP-1 in cerebral ischemia contributes to further understanding the pathophysiologic mechanism of stroke and finding a new therapeutic target.

Objective To explore the effects of microwave irradiation on the proliferation of keloid-derived fibroblasts so as to analyze the expression of collagen-1 and the activation of the signaling pathway involved.Methods Cells from a human keloid scar were cultured in vitro and randomly divided into a control group withont any intervention,a 10 mW/cm2 microwave irradiation (10-MI) group and a 20 mW/cm2 microwave irradiation (20-MI) group.Aliquots of the latter 2 groups were irradiated at their corresponding intensities for 5 min,15 min and 30 min.The growth of fibroblasts was evaluated using MTT assay.The expression of collagen-1 and changes in the phosphorylation of protein JNK were detected using western blotting.Results Compared with the control group,no significant differences in the average growth of the keloid-derived fibroblasts were observed in the 10-MI group,but significant differences were observed in the 20-MI group and among the three sub-groups irradiated for different durations.The expression of type 1 collagen was significantly down-regulated after irradiation in a time-dependent manner.After microwave radiation at 20 mW/cm2,JNK was significantly activated compared to the control group at the different time points.Conclusions Microwave irradiation at 20 mW/cm2 can significantly inhibit the proliferation of keloid-derived fibroblasts and the down-regulalion is correlated with the irradiation's duration.It can also significantly inhibit collagen-1 expression and relieve scar formation through activating the JNK signal pathway.

Objective To establish the RP-HPLC method for the determination of paeoniflorin in Cervipower Capsules. Methods Diamonsil~(TM)C_(18)column was used,with a mobile phase of A(methanol)-B(0.025 mol?L~(-1)disodium hydro- gen phosphate,adjusting pH being 7.4 with 0.025 mol?L~(-1)potassium dihydrogen phosphate),and with a gradient elu- tion,the flow rate was 1.0 mL?min~(-1),the detection wavelength was 230 nm,and the column temperature was 30℃. Result The linear range was 15.10～105.70?g?mL~(-1),r=0.999 9,and the average recovery for paeoniflorin was 100.1%,with RSD 1.94 %(n=9).Conclusion The method is accurate and reproducible,and can be used to deter- mine the content of paeoniflorin in Cervipower Capsules.

Objective:To investigate the effect of microRNA-155(miR-155) on transforming growth factor β2 (TGF-β2)-induced epithelial-mesenchymal transition of human retinal pigment epithelial cells and its mechanism.Methods:The retinal pigment epithelial cell ARPE-19 cell line was used as the research object.The cells cultured with DMEM medium were served as the control group and the cells cultured with DMEM medium containing 10 ng/ml TGF-β2 were served as the TGF-β2 group.The ARPE-19 cells transfected with miR-155 inhibitor were set as the miR-155 inhibitor group and the ARPE-19 cells transfected with miR-155 negative control were set as the miR-155 negative control group, and the cells in the two groups were cultured in DMEM medium containing 10 ng/ml TGF-β2.After 48 hours cell culture, reverse transcription-PCR was used to detect the expression of miR-155 in each group, and scratch migration test and Transwell chamber test were used to detect cell migration and invasion ability, and Western blot was used to detect the expressions of phosphate and tension homology deleted on chromosome ten gene (PTEN), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), p-Akt and epithelial mesenchymal markers E-cadherin (E-cad), zonula occludens protein 1 (ZO-1), F-actin, α-smooth muscle actin (α-SMA), fibronectin 1 (FN-l) vimentin, proteins.The target gene prediction library predicted miR-155 target gene and fluorescein enzyme reporter vectors were used to identify target genes.Results:After 48 hours of culture, the cells in the control group were in good condition with tight adherence and regular shape.The cells in the TGF-β2 group showed more obvious spindle shape with loose arrangement, and most of the cells were fibrous.The relative expression level of miR-155 in the cells of TGF-β2 group was 0.92±0.14, which was significantly higher than 0.35±0.06 of the control group ( t=7.242, P=0.003). The relative expression level of miR-155 in the cells of miR-155 inhibitor group was 0.21±0.03, which was significantly lower than 0.98±0.09 of the miR-155 negative control group ( t=12.421, P<0.01). The migration rate was higher and the number of cells passing through basement membrane was more in the TGF-β2 group than those of the control group, and the migration rate was higher and the number of cells passing through basement membrane of miR-155 was more in the miR-155 negative control group than those of the miR-155 inhibitor group, and the differences were statistically significant (all at P<0.01). Compared with the control group, the relative expression levels of PTEN, E-cad, ZO-1, F-actin protein were decreased, while the relative expression levels of PI3K and the p-Akt/Akt ratio were increased, and the relative expression levels of α-SMA, FN-1, vimentin proteins were increased in the TGF-β2 group, and the differences were statistically significant (all at P<0.01). Compared with the miR-155 negative control group, the relative expression levels of E-cad, ZO-1, F-actin and PTEN proteins were increased, while the relative expression levels of α-SMA, FN-l, vimentin, PI3K and the p-Akt/Akt ratio were decreased in the miR-155 inhibitor group, and the differences were statistically significant (all at P<0.01). Target gene prediction library prediction and luciferase reporter vector identification confirmed that PTEN was a downstream target gene of miR-155. Conclusions:miR-155 can promote the TGF-β2-induced epithelial-mesenchymal transition progress in human retinal pigment epithelial cells, and its mechanism may be related to inhibiting the expression of the target gene PTEN and stimulating the activation of the PI3K/Akt signaling pathway.

Objective To investigate the roles of Xenon-CT cerebral blood flow perfusion imaging in cerebral revascularization before surgery and efficacy evaluation.Methods The clinical data of 15 patients with symptomatic cerebral artery stenosis/occlusion of anterior circulation were analyzed retrospectively.Eight patients were treated with endovascular stenting,1 was treated with internal carotid endarterectomy,and 6 were treated with superficial temporal artery-middle cerebral artery bypass grafting.The regional cerebral blood flow (rCBF) detected by Xenon CT within 2 weeks before and after procedure and the modified Ranking scale (mRS) scores at 6 months after procedure were compared.Results (1) The mean rCBF value of 12 patients with abnormal blood perfusion of target vessels before procedure was 30±10 ml/(100 g·min) and that was 32±14 ml/(100 g·min) after procedure.Compared with before procedure,the difference was statistically significant (P<0.05).The mean rCBF value of 3 patients with normal blood perfusion of target vessels before procedure was 48±6 ml/(100 g·min) and that was 50±7 ml/(100 g·min) after procedure.Compared with before procedure,the difference was not statistically significant (P>0.05).(2) The postoperative mRS score was decreased in 8 cases and stable in 7 cases.Compared with before procedure,there were significant differences in mRS scores after procedure in 15 cases (P<0.05).During the follow-up period,none of the patients had new neurological impairment.Conclusion Revascularization can improve the presence of hemodynamic disorders in patients with symptomatic anterior circulation cerebral artery stenosis or occlusion of the target blood vessels in the distal local cerebral perfusion and neurological deficit symptoms.The patients with abnormal perfusion of preoperative Xenon-CT cerebral blood flow perfusion imaging may be more beneficial than those with normal perfusion.

Objective To investigate the relationship between intracranial atherosclerotic plaque stability and stroke recurrence risk.Methods Forty-eight patients with acute ischemic stroke caused by intracranial atherosclerosis in Guangdong Province Hospital of TCM were analyzed retrospectively.After the Essen Stroke Risk Scale (ESRS) was used to assess the risk factors for the patients,they were divided into either an ESRS ≥3 group (n=21 in the high-risk recurrence group) or an ESRS <3 group (n=27 in the low-risk recurrence group).Both groups of patients underwent high-resolution MR imaging (HR-MRI) examinations of the intracranial guilty vessels (basilar artery or unilateral middle cerebral artery).According to the signal intensity of HR-MRI on the T1-weighted imaging,T2-weighted imaging,and T1 fat suppression sequences,the intracranial atherosclerotic stable plaques and unstable plaques were distinguished.The stabilization of intracranial atherosclerotic plaques was compared in patients of both groups.Results There were significant differences in the age and incidences of hypertension,diabetes mellitus,and unstable plaques in patients of both groups (P<0.05).Further multivariate logistic regression analysis of the four factors showed that the age,hypertension,diabetes,and intracranial atherosclerotic unstable plaques were the high-risk factors for recurrent ischemic stroke (ORs,87.114,159.423,8.942,and 11.551,respectively;95%CIs 4.218-1 799.078,3.235-7 855.957,1.054-75.857,and 1.011-132.043,respectively;all P<0.05).Conclusion In addition to the traditional risk factors such as age,hypertension,and diabetes,the intracranial atherosclerotic unstable plaque is an independent risk factor for high-risk recurrence of ischemic stroke.

OBJECTIVE:To establish a method for the determining contents of 5 anthraquinones constituents in Sanhuang drop pills. METHODS:HPLC method was adopted. The column was Diamonsil C18 with the mobile phase of methanol-0.23% phosphoric acid(85∶15,V/V)at the flow rate of 1.0 ml/min,the detection wavelength was 440 nm and the volume temperature was room tem-perature. The sample size was 10 μl. RESULTS:The linear range was 0.070-0.700 μg for aloeemodin(r=0.999 9),0.646-6.460 μg for rhein(r=0.999 9),0.130-1.300 μg for emodin(r=0.999 9),0.150-1.500 μg for chrysophanol(r=0.999 9)and 0.074-0.740 μg for physcion(r=0.999 9),respectively. The RSDs of precision,stability and repeatability tests were no more than 1.96%. The av-erage recoveries were respectively 100.92%(RSD=2.80%,n=9),97.39%(RSD=1.39%,n=9),99.29%(RSD=1.81%,n=9), 100.73%(RSD=2.60%,n=9)and 99.81%(RSD=2.06%,n=9). CONCLUSIONS:The method is simple,accurate and specific, and can used for the content control of 5 constituents in Sanhuang drop pills.