简介：ToinvestigatetheexpressionlevelsofthreeDsbproteingenes,dsbB,dsbDanddsbG,atdifferenttimepointspostC.trachomatisinfection,mousefibroblast12cellswerechosentobeinfectedwithC.trachomatisserovarFstrainF/IC-Cal-13.C.trachomatiselementarybody(EB)-infected12cellswereharvestedimmediatelyafterEBattachmentontothecellsandevery4hourspostinfection(hpi)till44hpifortotalRNApreparation.RT-PCRassayswerethenemployedtoamplifycDNAwithprimerpairswhicharespecifictoC.trachomatisdsbgenesdsbB,dsbD,dsbGandtufArespectively.Therela-tiveexpressionlevelsofDsbproteingeneswereevaluatedascDNAratiosofgenedsbtogenetufA.OurresultsshowedthatthetranscriptionofdsbGstartedfrom12hpiandgraduallyincreasedtill44hpi.ThetranscriptionofdsbBanddsbDweredetectedat16hpiandreachedtheirpeaksat28hpiand24-28hpi,respectively.Moreover,therewasobvioustranscriptionofdsbBatthelaterstage(44hpi),butnonefordsbDatthistimepoint.WecametotheconclusionthattheexpressionlevelsofthethreeDsbproteingenesaredifferentduringthedevelopmentalcycleofC.trachomatist.Theymayplayaroleinmid-to-latestageofthedevelopmentalcycleofC.trachomatis.

简介：Thehousedustmites(Dermatophagoidesfarinae,Derf)arethemajorsourceofaeroallergensimplicatedintheexpressionofatopicdisorders,includingasthma,allergicrhinitisandatopicdermatitis.Inparticular,strongcircumstantialevidencesuggeststhathousedustmiteantigensareimportantprecipitatingfactorsofasthma.Manyhousedustmiteallergensareproteasesthatcanelicitairwayinflammationbystimulatingthereleaseofcytokinesfrombronchialepithelialcells.ToinvestigatewhetherDerfallergenproteasesinducedcytokineproductionfromtheepithelialcelllineBEAS-2B,BEAS-2Bcellswereculturedwith4differentconcentrationsofDerf(0.02,0.2,2,20μg/ml)for24-96h,afterwhichsupernatantswereassayedforinterleukin(IL)-6andIL-8withELISA.Reversetranscription-PCRwasalsoperformed.ThecellsheetswereintactthroughouttheobservationincontrolgroupwithoutanyexposuretoDerfantigen.IntheexperimentalgroupscellstreatedwithDerfallergenshowedchangesintheanchoragestatusofthemonolayer.Therewasasignificantincreaseinthelevelofcytokineproductioncomparedwiththeuntreatedsample.ThereleaseofIL-6andIL-8increasedinaconcentration-dependentmanner(P<0.05,respectively)withtheadditionofincreasingdosageofDerftothecellsheets.LevelsofIL-6andIL-8begantoriseat24hand48hafterallergenexposure,andtheyincreasedsignificantlyinthesupematantsat72hand96h.AtthesametimetheconcentrationdependenceofinductionofIL-6andIL-8expressionaswellasanincreaseintheexpressionofIL-6andIL-8mRNAmanifestedevidently.HDM-inducedairwayinflammationmayincludeDerf-mediatedreleaseofinflammatorymediators,andtheproteolyticactivityofanallergenmaystimulatethereleaseofproinflammatorycytokinesfromhumanbronchialepithelium.ItissuggestedthatIL-6andIL-8productionbybronchialepithelialcellsmayplayaroleinthepathogenesisofallergicasthma.