Objective: To construct the recombinant plasmid targeting M2 gene of respiratory syncystial virus(RSV) for further study of RNA interference(RNAi) technology in inhibiting RSV infection.Methods: According to the designing principles of shRNA,The M2 gene of RSV was selected as the target gene.A 19bp reverse repeated sequences with 9bp spacer were designed and synthesized.The complement strain were obtained by annealing and inserted into the vector pgenesil-1 containing green fluorescenet protein(GFP) sequence and U6 promotor.Finally the recombinant plasmid were identified by enzyme digesion and DNA sequencing,then transfected into HEP2 cells and observed by fluorescence microscope.Results: The recombinante plasmid targeting the mRNA of RSV M2 gene was successfully constructed and the good efficents of transfection were found by fluorescence microscope.Conclusion: The successful construction of the pshRNA recombinant plasmid targeting the M2 gene of RSV will help futher study on application of the RNAi technology to anti-RSV infection.

OBJECTIVE: To discuss the competitive intelligence(CI) development strategies in Chinese pharmaceutical enterprises. METHODS: The CI in Chinese pharmaceutical enterprises was overviewed and the problems were analyzed. Then development strategies of CI were put forward. RESULTS & CONCLUSION:CI is one of the powerful competitive tools in pharmaceutical enterprises; development strategies of which should be further explored.

OBJECTIVE:To provide retrieval techniques for chemical and medical workers to mater the latest technology dynamic state.METHODS:The four basic retrieval methods of CA on CD(American Chemical Abstracts)were introduced,and of which,several classic logical combination retrieval techniques were exemplified in detail.RESULTS & CONCLUSIONS:The retrieval of CA on CD is characterized by convenience,rapidity,flexibility,multiform logical combination.Medical information worker should skillfully use the combination retrieval techniques and unceasingly explore the exemplification of these retrieval techniques.

Objective To elucidate the airway inflammation status in mice infected with respiratory syncytial virus (RSV), and whether the inflammation could be alleviated by small interference RNA (siRNA) targeting specific RSV gene. MethodsBALB/c mice were infected with RSV via intranasal instillation of RSV suspension, and were then treated with specific siRNA targeting RSV-M2 gene. ELISA assay was used to detect the levels of IFN-γ, IL-12 and IL-10 and Microscope was used to count white blood cells in bronchoalveolar lavage fluid(BALF). ResultsAfter RSV infection, a significant increase in leukocytes count was observed in BALF. Differential count showed a rise in the percentage of neutrophils, eosinophil, especially lymphocytes and a reduction of the percentage of monocytes and macrophages( P＜0.05 ).The levels of IFN-γ, IL-12 and IL-10 were also increased(P＜0.05). Furthermore, the leukocytes count,the percentage of lymphocytes, neutrophils and eosinophil, and the levels of IFN-γ, IL-12 and IL-10in BALF were decreased accordingly while the mice were given higher concentrations of siRNA ( P＜0.05 ).Conclusion RSV caused airway inflammation in BALB/c mice, which may be alleviated by RNAi technology.

Objective To compare the dosemetry between helical tomotherapy (HT) and intensity-modulated radiotherapy (IMRT) for early-stage postoperative breast cancer and provide more valuable evidences to the clinical researches. Methods Clinical trails of dosimetric comparing between HT and IMRT for early-stage breast cancer were obtained from PubMed, Embase, Sciencedirect, CNKI, VIP and Wanfang databases, evaluated and analyzed with the Cochrane Collaboration's RevMan 5.2 software. Results 10 studies were included with a total of 135 patients. Compared to IMRT plans, HT plans provided a significantly better conformity index (P<0.000 1), mean (P<0.000 01) and maximal dose (P=0.003) of the planning target volume (PTV). HT plans had a lower heart maximal dose (P=0.005), V20 (P=0.05), V30 (P=0.003), and ipsilateral lung maximal dose (P=0.003), V20 (P=0.02), as while as had a higher contralateral breast V5 (P=0.01), mean (P=0.05) and maximal dose (P<0.000 01). There was no significantly difference between HT and IMRT plans for homogeneity index of PTV, heart V5, V10, mean dose, ipsilateral lung V5, V10, V30, mean dose, contralateral breast V10, contralateral lung mean and maximal dose (all P >0.05). Conclusion Compared to IMRT plans, HT plans have the dosimetry superiority for early-stage breast cancer with significantly better coverage and dose conformity while maintaining lower doses to high risk organs.

Objective To observe the effect of curcumin on the expression of smad7 and TGF-β1 in rats renal tubular cells of with Ang II, and discuss the mechanism of curcumin to improve renal interstitial fibrosis.Methods Cultured rat renal tubular epithelial cells were divided into the blank group, the Ang II control group and the low, medium and high dose curcumin group. The rest of the groups were intervened by 10-8 mol/L Ang II except the blank group; the low, medium and high dose groups of curcumin were intervened by 2.5, 5.0, 10.0μmol/L curcumin. Then Western blot was used to detect the expression of TGF-β1 and smad7 protein, RT-PCR was used to detect the TGF-β1 and smad7 mRNA expression.Results Compared with the blank group, the expression of TGF-β1 protein (0.23 ± 0.03vs. 0.16 ± 0.01), and TGF-β1 mRNA (1.89 ± 0.20vs. 1.00 ± 0.00) significantly increasedin AngⅡ control group (P<0.05), and the expression of smad7 protein (0.19 ± 0.03vs. 0.24 ± 0.02), and smad7 mRNA (0.48 ± 0.05vs. 1.00 ± 0.00) significantly reduced in AngⅡcontrol group (P<0.05). Compared with the AngⅡ control group, the expression of TGF-β1 protein in low, medium and high dose curcumingroup (0.18 ± 0.02, 0.17 ± 0.02, 0.16 ± 0.03vs. 0.23 ± 0.03) and TGF-β1 mRNA (1.58 ± 0.11, 1.34 ± 0.16, 0.97 ± 0.19vs. 1.89 ± 0.20) significantly decreased (P<0.05), and the expression of smad7 protein (0.28 ± 0.04, 0.31 ± 0.03, 0.34 ± 0.04vs. 0.19 ± 0.03) and smad7 mRNA (0.68 ± 0.07, 0.80 ± 0.06, 0.98 ± 0.09vs.0.48 ± 0.05) increased significantly (P<0.05).Conclusions Curcumin can thus play its role in renal protection by counteract the AngⅡ mediated renal interstitial fibrosis. Its mechanism may be related to the reduction of TGF-β1 protein and its mRNA expression, up regulation of smad7 protein and its mRNA expression.

Objective To investigate the protective effects of folic acid on the oxidative damage that ox-LDL (oxi?dized low-density lipoprotein receptor 1) render to human umbilical vein endothelial cells (HUVEC). Methods HUVECs were injured by ox-LDL (120 mg/L) for 24 h while they were incubated with various concentration of folic acid (0,15, 60, 150, 225, 300, 375 nmol/L). Then HUVECs were cultured in media contains same concentration of folic acid but without ox-LDL for 72 hours. Finally, HUVECs were harvested after 24, 48, 72 and 96 h. The morphological changes were observed us?ing inverted microscope and cell viability were examined by MTT. Results Various concentrations of folic acid (0,15, 50, 100, 200 and 500 nmol/L) has no obvious promotion or inhibition effect in growth of normal HUVEC (P>0.05). However, compared with the ox-FA-def group, 150, 225, 300 and 375 nmol/L of folic acid promoted proliferation of HUVECs with 96 and 120 hours of incubations (P < 0.05). Folic acid of 60, 150, 225, 300 and 375 nmol/L promoted the proliferation of HUVECs with 72 h and 96 hours of incubation (P<0.05). Conclusion High dose folic acid can reduce the ox-LDL oxida?tive damage on HUVEC in a concentration dependent manner.

Objective To explore the effect of arsenic trioxide (As2O3) on the migration of neurons and the potential mechanism through the establishment of primary neuron culture from the brains of neonatal rats.Methods Brain tissues were selected from SD neonatal rats for primary neuron calture.The cells were divided into 4 groups based on the addition of As2 O3:normal control group,1 μmol/L As2O3 group,10 μmol/L As2O3 group and 20 μmol/L As2O3 group.The primary neurons were treated with different concentrations of As2O3 and cultured for 24 hours.Boyden chamber assay was used to detect the effect of As2O3 on neuronal migration.Immunofluorescence laser confocal microscope was used to observe the structure of actin.Results In the control group,the cultured neurons showed a regular pattern of distribution.In the 3 groups treated with As2O3,the distribution of neurons was loose and disordered,which was most obvious in the 20 μmol/L As2O3 group.The results showed that the higher concentration of As2O3,more difficult it was for the neurons to survive.The number of neuronal migration was 64.6 ± 4.3 for normal control group,63.0 ± 7.0 for 1 μmol/L As2O3 group,54.8 ± 3.6 for 10 μmol/L As2O3 group,and 21.6 ± 3.9 for 20 μmol/L As2O3 group.The results showed that As2O3 might inhibit the migration of primary neurons in a dose-dependent manner (F =49.31,P ＜0.001).The normal actin skeleton was destroyed under the laser confocal microscope in 10 μmol/L As2O3 group and 20 μmol/L As2O3 group,while they remained unaffected in normal control group and 1 μmol/L As2O2 group.Conclusion As2 O3 exposure can reduce the neuron migration in a dose-independent manner probably through disrupting the organization of acting cytoskeleton.

Objective To explore the effects of arsenic exposure on learning and memory and its potential mechanism in rats.Methods Water-based arsenic-exposed rat models were established on 4-l0 postnatal days.The experimental animals were divided into 4 groups (10-12 cases in each group):the control group,the 15 μg/L As2O3 water group,the 30 μg/L As2O3 water group,and the 45 μg/L As2O3 water group.Cognitive functions were examined with the Morris water maze,exploratory behavior was detected by the exploratory behavior test.The hippocampus of pups from each experimental group was sectioned at various time points after arsenic exposure.The morphologies and neurogenesis of the neurons in the hippocampus CA1-CA3 region and dentate gyrus (DG) were observed by hematoxylin-eosin staining,Nissl staining,and doublecortin (DCX) immunostaining at different time points after arsenic exposure.Results Compared with the normal control group,the escape latency of the rats in the arsenic-exposed group was prolonged.The average escape latency of the rats in the normal control group,15 μg/L As2O3 group,30 μg/L As2O3 group and45 μg/L As2O3 group were (17.00±9.53) s,(35.89 ±19.81) s,(26.60 ±18.84) s,and (33.79 ±18.08) s,respectively,and the difference among 4 groups was statistically significant (F =3.591,P ＜ 0.05),and the residence time in the original target quadrant was shortened,respectively,(38.93 ± 8.33) s,(36.03 ± 16.25) s,(29.85 ± 9.27) s,and (29.84 ± 10.16) s,respectively,and there was no significant difference among 4 groups (F =1.681,P =0.187).HE staining and Nissl staining showed that pathological changes such as edema,degeneration and necrosis were observed in the hippocampal CA1 area and CA2 area as well as dentate gyrus cells in rats exposed to arsenic in the acute phase.The higher the concentration of arsenic exposure,the more obvious the cell structure disorder was.However,5 weeks after exposure,the pathological changes in hippocampal neurons in the arsenic-exposed group gradually returned to normal.Immunohistochemistry showed that the expressions of DCX in the CA1,CA2 and dentate gyrus of rats exposed to arsenic decreased significantly 24 h after arsenic exposure,especially in the 45 μg/L group.Five weeks after arsenic exposure,there was no expression in the hippocampal CA1-CA3 area,and there was still a small amount of expression in the dentate gyrus.Conclusions Postnatal low-concentration arsenic exposure may impair learning and abnormal germination of neurons in the hippocampal dentate gyrus may be the underlying mechanism.

Allergen specific immunotherapy（AIT）is currently the only recognized treatment for the cause of allergic diseases.With the gradual increase in the dose of allergens，AIT can induce the body to establish immune tolerance，prevent the aggravation of allergic symptoms，reduce patients' emergency medication，and block the development of allergic diseases.AIT effectiveness and safety have been affirmed by scholars at home and abroad.Successful AIT can inhibit early and late anaphylaxis，but not all patients respond to AIT.The overall response rate ranges from 50% to 80%，and the course of treatment is long and the cost is high.Therefore，it is particularly important to identify biomarkers that can predict and evaluate the efficacy.This article mainly reviews the research progress on the correlation between the changes of biomarkers（immunoglobulin and cytokines）and the efficacy of AIT，in order to provide reference for pediatricians.