Study1the Research Of YAP Protein Expression In Bronchial Smooth Muscle Of Asthma Study2the Preliminary Explore The Methods Of The Rapid Detection Of Legionella Pneumophila

Objective:Through animal experiments, establishment of mouse models of chronicbronchial asthma,Exploring if the normal mice or asthmatic mice could expres theYAP (Yes-assoeiated protein) protein in bronchial smooth muscle.In order to providenew targets for the treatment of bronchial asthma.Methods:Select40BALB/C mices of6-8weeks old、healthy、female、SPF level,The40mices are randomly divided into experimental and control groups,each group has20mices,the experimental group is the group of chronic asthma model.Asthma model mice:each mouse is injected intraperitoneally on days of0,7,14with0.2ml allergens liquid,in the beginning of21day all mice were placed inthe sealed box of homemade,application of compressed air atomizer for atomizerstimulate with OVA,conducted three times a week, every time30min,continuousatomization eight weeks;Normal control mice:given the same dose of saline instead of OVA for sensitization+excitation.Symptoms were observed in all mices,after24h of the last challenge retinalartery bleeding, mice were sacrificed by cervical dislocation,recycling blood,test thetotal IgE and OVA-specific IgE levels in the serum;take the left lung,HE staining beused to observe pathological changes in the lung tissue;use immunohistochemical todetecte whether it have YAP protein expression in the lung tissue..Results:1.symptoms of the two groups of mice:after atomization challenge,asthmaticmice,symptoms:shortness of breath,irritability, scratching,hair lodging,urine increased,finally fell down and not moving and so on,the time of the symptoms beganing toappear will be getting shorter, and duration longer;normal mice are relatively quiet.2.Compared with the normal control group, the levels of the total IgE and theOVA-specific IgE in serum are all higher for the asthma model group (P <0.01),thedifference is statistically significant.3.The HE staining results of asthmatic mice lung tissue:there are manyinflammatory cell infiltration around bronchial and perivascular,in which showseosinophils and lymphocytes,airway smooth muscle proliferation and other changes.No obvious abnormalities pathological changes in lung tissue of mice in the controlgroup.4.The control group almost have noYAP protein expression;of the modelgroup,in some inflammatory cells have YAP protein expression,but the expression isweak, have no statistical significance.Conclusion:1.The levels of the total IgE and the OVA-specific IgE in the serum are all higherfor the asthma model group than control group;The result of HE staining of lungtissue is compliance with asthma management change.Therefore,the modeling of theexperimental methods used is correctly,so it can be used in the present study;2.Normal lung tissue of mice without the expression of YAP protein;3.In some inflammatory cells there have YAP protein expression of the modelgroup,but the expression is so weak that it have no statistical significance.So there is no significant expression of YAP protein in bronchial smooth muscle. ObjectiveUnder the existing technical conditions,explore a fast and accurate method forthe detection of Legionella pneumophila(LP) under simple experimental conditions.In order to provide a new way of thinking which can quickly clear the pathogens ofLegionella pneumophila.MethodsDesign and synthesis the primers using for gene amplification of the standardDNAfragments of Legionella pneumophila;Using loop-mediated isothermal nucleic acid amplification technology(loop-mediated isothermal amplification,LAMP) and PCR techniques to amplifyconserved DNA fragments of Legionella pneumophila and other controlstrains,verified the specificity of LAMP reaction and sensitivity;to verify thespecificity and sensitivity of LAMP reaction;The LAMP amplification product hybridization with the DNA microarray,whether the observed hybridization signals can occur in the corresponding region ofthe chip,to detect the specific of LAMP technologies with gene chip technology in thedetection of Legionella pneumophila.Resultsafter LAMP reaction only the LP DNA has corresponding amplifiedbands;compared to the minimum dilution factor of105of PCR,the LAMP can dilutionup to107,so the sensitivity of the reaction is about100times higher than PCR.Afterthe hybridization of LAMP amplified genes and gene chip,the corresponding LP areaoccur hybridization signal.Conclusion 1.Because only the LAMP productions of Legionella pneumophila have specificamplification bands,so the LAMP technique has a high specificity in the amplificationof the corresponding gene;2.Compared to ordinary PCR technique,the sensitivity of LAMP technology isabout100times more than the PCR,hence the LAMP technology with highsensitivity;3.After hybridization,hybridization signals can occur in the area of theLegionella pneumophila,the other areas have no,so LAMP technology combines withgene chip technology has a high specificity in the detection of Legionellapneumophila.