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 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.

The peptidyl-prolyl imide bond cis/trans isomerization of Xaa-Pro motif in the peptide and protein plays an important role to influence their conformation and function. Here, a series of model peptides including phosphorylated and its unphosphorylated counterparts were designed and synthesized. Preliminary 1H NMR experiments and molecular dynamics (MD) simulation were used to analyze the peptidyl-prolyl cis/trans imide bond isomerization. The data indicated that the side-chain O-phosphorylation of the Xaa residues preceding proline affected evidently the isomerization and thereby regulated the peptides conformations. The charges of the phosphate moiety as well as their steric effects might be the driving force for the conformational changes of these phosphopeptides.Moreover, the obtained most stable multiple configurations and their statistic cis/trans concentration distribution in MD simulation were basically consistent with the NMR experiments, which demonstrated that phosphorylation increased the cis conformation of the peptide and the maximum cis ratio is given while the phosphate group has no negative charge.

Objective:To investigate the complications of traumatic optic neuropathy and to call the surgeons' attention to precaution. Method:Retrospectively analysis of 122 cases patients with traumatic optic neuropathy and 3 cases were analyzed in detail including 1 case with purulent meningitis, another 1 case with internal carotid artery pseudoaneurysm and the other 1 case with internal carotid artery cavernous sinus fistula. Result:Most of the patients had the complications of orbital fracture, maxillofacial fracture, ocular and craniocerebral injury. A few of patients had other injuries all over the body. The case with purulent meningitis was cured with antibiotics. The case with internal carotid artery pseudoaneurysm was cured with neurosurgery. The visual acuity of the both cases were improved. The case with internal carotid artery cavernous sinus fistula died of severe hemorrhea. Conclusion: The patient with traumatic optic neuropathy has the possibility of severe cranial disorders,orbital fracture, maxillofacial fracture and injuries of viscera or limbs. It should be paid mare attention and treated accordingly.

Objective To investigate the relationship of S100B protein expression and the pathogenesis of early-onset and late-onset preeclampsia.Methods Sixty patients with preeclampsia who received caesarean section at Qingdao Municipal Hospital from October 2010 to September 2011 were enrolled in this study.Thirty cases were early-onset preeclampsia( referred as early-onset preeclampsia group,＜ 34 weeks),and the other 30 cases were late-onset preeclampsia (referred as late-onset preeclampsia group,≥34 weeks).Thirty women who received caesarean section because of pelvic structural deformities,breech presentation,macrosomia and social factors were included as the control group.The expression of S100B mRNA in the placenta was detected by reverse transcription ( RT)-PCR.The expression of S100B protein in the placenta was detected by immunohistochemistry.Results ( 1 ) S100B mRNA was expressed in the trophoblasts of preeclampsia and control groups.The expression of S100B mRNA in early-onset preeclampsia group (0.73 ±0.11 ) was significantly higher than the control group (0.58 ±0.08) and lateonset preeclampsia group (0.64 ±0.10,P ＜0.05 ).There was no significant difference between late-onset preeclampsia group and the control group ( P ＞ 0.05 ).(2) S100B protein was expressed in the plasma membrane and cytoplasm of the trophoblasts,correlated positively with the brownish yellow and brown particles inside the cells.It was expressed in all the three groups.Immunohistochemistry revealed that the expression of S100B protein in the placenta of early-onset preeclampsia group was 100％ (30/30),significantly higher than those of late-onset preeclampsia group and the control group,in which the positive rate were 70％ (21/30) and 63％ (19/30) respectively (P ＜0.05).There was no difference between late onset preeclampsia group and the control group (P ＞0.05).Conclusion Early-onset and late-onset preeclampsia may have different etiology and pathogenesis.S100B may be a factor in the pathogenesis of early-onset preeclampsia.

Objective To retrospectively review the efficacy and side effects of intense pulsed light (IPL) and red light emitting diode (LED) in the treatment of steroid-dependent dermatitis.Methods Seventy patients with steroid-dependent dermatitis mainly manifesting as facial telangiectasis were treated with IPL for an average of 3.49 sessions with a 4-week interval.The energy density of IPL varied from 20 to 23 J/cm2,pulse width from 2.6 to 5.0 ms,and delay from 15 to 20 ms.Meantime,197 patients with steroid-dependent dermatitis,who mainly presented with facial skin sensitivity,were treated with red LED (633 ± 3 nm wave length) twice a week for an average of 4.23 sessions.The energy density of red LED was 128 J/cm2,and the irradiation lasted 20 minutes at each treatment.The efficacy and adverse reactions were assessed and recorded for each treatment.Results The total response rate was 88.57％ for IPL,and 83.76％ for red LED.There was a significant difference in the clinical efficacy between triple-pulse and double-pulse IPL (x2 =8.14,P ＜ 0.05).No severe adverse reaction was observed in any of the patients.Conclusion IPL and red LED are both effective in treating steroid-dependent dermatitis.