简介：Humanp100proteinconsistsoffourrepeateddomainsofstaphylococcalnuclease(SN)-likedomain,aswellasatudor(TD)domainthereafter.WehavepreviouslyshownthattheSN-likedomainofp100interactedwithSTAT6andthelargesubunitofRNApolII,resultingintheenhancementofSTAT6-mediatedgenetranscriptionalactivation.Here,weshowthatSN-likedomainalsointeractedwithCREBbindingprotein(CBP)anddirectlyenhancedtheacetyltransferaseactivityofCBPonhistone.Ontheotherhand,overexpressionofCBPalonehadnoabilitytosignificantlyincreaseSTAT6dependenttranscriptionalactivation,however,togetherwithp100protein,sufficientlyenhancedtheactivationoftranscriptionwhichwasinlinewiththepreviousresultthatp100proteinbridgedSTAT6withCBP.

简介：Thepurposeofthissubjectwastoinvestigatemolecularepidemiologyofoxacillin-resistantStaphylococcusaureus(MRSA)isolatedfromhospitalizedpatients,andtosurveytheinvitroactivityofteicoplanin,vancomycinandother9antibioticsagainstStaphylococcusspecies.MRSAweredetectedbyoxacillin-NaCl-containingscreenagar.ThehomologyofnosocomialMRSAfromICUandRCUwasdeterminedbypulse-fieldgelelectrophoresis.Agardiffusion,EtestandagardilutionwereusedtocomparetheinvitroactivityofteicoplaninandvancomycinagainstStaphylococcussppfrom2001to2003atPekingUnionMedicalCollegeHospital.WHONET-5.3softwarewasusedtoanalyzetheantimicrobialsusceptibilitydata.From2001to2003,theprevalencesofMRSAwere56.5%,65.3%,64.7%,respectively.PFGEfoundmostofMRSAfromICUandRCUwerecloselyrelated.AllofS.aureusandS.epidimidisisolatesweresusceptibletoteicoplaninandvancomycinfrom2001to2003.However,1isolateofS.haemolyticuswasresistantand9isolatesintermediatetoteicoplanin.Theminimalinhibitoryconcentrationofteicoplanindidnotcorrelatewellwithzonediameter,wheninoculumincreasedby100folds,thezonediametersofteicoplanindecreasedmoregreatlythanthoseofvancomycin.In2002,severeoutbreakscausedbyMRSAstrainshadbeenfoundinICUandRCUwards.TeicoplaninandvancomycinhadgoodactivityagainstclinicalisolatesofStaphylococcispp.TeicoplaninwaslessactivethanvancomycinagainstS.haemolyticus.MostofS.haemolyticusisolateswereintermediatetoteicoplanin.Antimicrobialsusceptibilitytestingofteicoplaninwasinfluencedbythediffusionspeedintheagarandinoculumsize.

简介：InordertoimprovethefunctionalaffinityofthehumanizedVHsingledomainantibodyagainsthumanlungcancer,thegenescodingthehomogenousdimersdihu3D3V_Handtetramerstehu3D3V_HwereconstructedbyfusingtheSVS-Cysshortpeptideandp53tetramerizationstructuraldomaingenetohu3D3V_HgeneviarecombinantPCRtechnique,respectively.Then,thedihu3D3V_Handtehu3D3V_HgeneswereclonedtotheprokaryoticexpressionvectorpET-22b(+)andexpressedinE.coliBL21(DE3).TheproteinsexpressedwerepurifiedthroughNi~(2+)-affinitychromatographiccolumn.Meanwhile,thehu3D3V_H,dihu3D3V_Handtehu3D3V_HproteinswerelabeledwithFITC,andtheirreactivitywithan-tigenandspecificitywereanalyzedbyimmunofluorescenceassay.Astotheirfunctionalaffinities,itwasanalyzedandcomparedbyflowcytometry.Theresultsindicatedthatthesetwogeneswereexpressedasmonomersandmainlyasinclusionbodies.Afterpurificationandrenaturation,therewereabout50%ofdimersand70%oftetramerremainingintheproteinsolution.Inaddition,thedihu3D3V_Handte-hu3D3V_Hproteinsstillremainedthereactivitywithantigenandspecificityofhu3D3V_Hprotein,andtheirfunctionalaffinitieswereincreasedabout60%or100%respectively,comparedwiththoseofhu3D3V_Hprotein.Itisevidentthatthefunctionalaffinityofhu3D3V_Hproteincanbegreatlyimprovedbyincreasingitsbindingvalency.

简介：Toinvestigatetheinfluenceofmycophenolatemofetil(MMF)uponthematurationandtheallo-stimulatoryactivityofculturedprogenitorsofdendriticcells(DCp),andtoevaluatetheeffectsofthepre-treateddentriticcellsofrecipientswithMMFonthetoleranceinductionaswellasitspossiblemechanism,GM-CSFandMMFwereaddedtotheinvitroculturedprogenitorcells,andtheimmuno-phenotypicalanalysiswasperformedbymeansofflowcytometry.ThesecretionofIL-12wasdetectedbyELISAandthestimulatoryactivitiesofDCponallogeneicTcellswereobservedbymixedlymphocytereaction.Twenty-fourC57BL/6miceweredividedinto3groups(eachwith8mice),inwhichgroupAofmiceacceptedallograftsofheartfromBALB/cmice,groupBofmicehadreceiveduntreatedDCpfromdonorsofBALB/cmice7daysbeforetransplantation,andC57BL/6miceingroupCweretreatedbyinjectionwithMMF-treatedallograftsofheartfromBALB/cmice7daysbeforetransplantation.Thesurvivaltimesofallograftsandthechangesofthecytokinelevelsinsereoftherecipientmicewereobservedaftertransplantation.TheexperimentalresultsshowedthatMMFcouldsignificantlyinhibittheexpressionsofthecostimulatorymoleculesCD80andCD86onDCsandthesecretionofIL-12andtheallo-stimulatoryactivitiesofDCswerealsomarkedlyinhibited.ThesurvivaltimesofallograftsingroupBofmicewerelongerthanthoseingroupA,whilethegroupCshowedthelongestsurvivaltimesofallografts,withamarkedreductionintheproductionoftheThltypecytokines.ItisevidentthatMMFhasasuppressiveeffectonthematurationandallo-stimulatoryactivitiesofthecultureddendriticcellprogenitors,thusleadingtoadonorspecifictoleranceinheart-transplantedrecipients.

简介：Toclonethegenecodingtheimmunodominantregioninthechlamydialprotease-likeactivityfactor(CPAF)fromChlamydophilapneumoniae,toanalyzeimmunocompetenceoftheexpressedprotein,andtoevaluateitsvalueinserodiagnosis,theCPAFimmunodominantregiongenewasamplified,ligatedintoapGEX6p-2vector,andthentheexpressedrecombinantproteinwaspurifiedwithglutathioneS-transferase(GST)agarosegelFFafterrenaturation,thenidentifiedbySDS-PAGEandWesternblot.AnewindirectELISAwasdevelopedwiththepurifiedproteinascoatingantigen.TheimmunogenicityoftherecombinantproteinwasevaluatedbyimmunizationtoNewZealandrabbits,anditsimmunoreactivitywasanalyzedbyreactingwithanti-C,pneumoniaeantibody.300clinicalserasampleswererespectivelyde-tectedbymicroimmunofluorescence(MIF)asreferencemethodandtheindirectELISA,andthediffer-encebetweenthetwomethodswasanalyzed.Cross-reactivityagainstChlamydiatrachomatiswasinvesti-gatedwiththeindirectELISAtodetectanti-C,trachomatispositiveantisera.Theresultsindicatedthata51.3kDarecombinantproteinwasobtained.Westernblotassayprovedthattherecombinantproteincouldmerelyspecificallyreactwithhumananti-C.pneumoniaeantisera.ThetitersofthespecificIgGan-tibodiesintheimmunizedNewZealandrabbitswereabove1:16000.Anti-C.pneumoniaeIgGpositiveandnegativereferencesereweredetectedwiththeindirectELISA,andtheconcordancerateofnegativeandpositiveresultswereboth100%(40/40).ThesensitivityandspecificityoftheindirectELISAincomparisonwithMIFwere93.8%(45/48)and100%(252/252)separatelybydetecting300clinicalserasamples,andtheconcordanceratebetweenthetwomethodswas99.0%.NocrossreactionagainstC.trachomatiswasfoundwiththeindirectELISAtodetectanti-C,trachomatispositiveantisera.Incon-clusion,thepreparedrecombinantproteinoftheCPAFimmunodominantregionshowsexcellentimmuno-competenceandcanbeusedtodevelopanewindirect

简介：Wehaveconfirmedefficientanti-tumoractivitiesoftheperipherallymphocytestransducedwithap185HER2-specificchimericT-cellreceptorgenebothinmurineandinhumaninourpreviousstudies.TofurthertestthefeasibilityofchimericT-cellreceptorinabonemarrowtransplantationmodel,wefirst,madetwomurinetumorcelllines:MT901andMCA-205,toexpresshumanp185HER2byretroviralgenetransduction.MurinebonemarrowcellswereretrovirallytransducedtoexpressthechimericT-cellreceptorandgene-modifiedbonemarrowcellsweretransplantedintolethallyirradiatedmouse.Sixmonthsposttransplantation,p185HER2-positivetumorcells:MT-901/HER2orMCA-205/HER2wassubcutaneouslyorintravenouslyinjectedtomakemousemodelssimulatingprimarybreastcancerorpulmonarymetastasis.Theinvivoanti-tumoreffectsweremonitoredbythesizeofthesubcutaneoustumororcountingthetumornodulesinthelungsafterIndiainkstaining.ThesizeofthesubcutaneoustumorwassignificantlyinhibitedandthenumberofpulmonarynodulesweresignificantlydecreasedinmouserecipientstransplantedwithchimericT-cellreceptormodifiedbonemarrowcellscomparedwiththecontrolgroup.Ourresultssuggesttheefficientinvivoanti-tumoractivitiesofchimericT-cellreceptorgenemodifiedbonemarrowcells.