简介：InordertoobserveseveralantibodiestoliverantigensinChinesepatientswithdifferentfiverdiseasesandtodiscussthecharacteristicsoftheautoantibodiesinautoimmuneliverdiseases,from1412patients,detectedbyindirectimmumofluoreseence(IIF)initially,230patientswithabnormalALTwerechosenanddividedinto5groups:①autoimmunediseasesgroup,42cases:18withautoimmtmehepatitis(AIH),21withprimarybiliarycirrhosis(PBC),3withprimarysclerosingcholangitis(PSC).②HAVgroup,23cases;③HBVgroup,70cases;④HCVgroup,35casesand⑤NonA-Egroup,60cases.First,ANA,AMA,SMA,liver-kidneymicrosomalantibody(LKM)andsoonweretestedby1/F.Then,LKM-1,fivercytosofic-1(LC-1),solubleliverantigen/fiverpancreas(SLA/LP)andsubtypeofAMA(M2)aswellasANAprofilesuchasSS-A,SS-BanddsDNAweretestedbyWesternblotandimmtmoblotstripsassay,respectively.Theresultswerethatamong1412cases,thosediagnosedasAIH,PBCandPSCaccotmtedfor12.7‰,14.9‰and2.1‰,respectively,ofthesamplesbeingtested.2/230withLKM-1and2/230withSLA/LPwereseeninindividualsinfectedwithAIHandHCV,respectively.AllpatientswithPBCshowedAMAandM2antibodies.NospecificANApatternwasseeninAIHby1/Fbutanti-actinwasonlyfoundinpatientswithAIH.InNonA-Egroup,fourcaseswerepositiveofAMAandM2;threehadhightiterofSMAandother4hadSS-A,SS-BordsDNAantibodies,etc.Itwasconcludedthatthedetectionofanti-fiverantigens,ANAprofileandAMAsubtypeswerehelpfulforthediagnosisofautoimmunefiverdiseasesandoverlapsyndromes.InpatientswithNonA-Ehepatitis,thediagnosisofPBCorAIHshouldbetakenintoconsideration.

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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.