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首页 Argiope spider venom extraction and preparation

Argiope spider venom extraction and preparation.doc

Argiope spider venom extraction…

李展照
2018-04-30 0人阅读 举报 0 0 暂无简介

简介:本文档为《Argiope spider venom extraction and preparationdoc》,可适用于高中教育领域

ArgiopespidervenomextractionandpreparationArgiopespidervenomextractionandpreparationOf:Chenwithwater,ZhangHao,YuanFenghui,MaoyueXiang,CaoNing,ZhuMangAbstractObjectiveTounderstandthelivinghabitsofthespiderArgiopeandmorphologicalcharacteristicsofthevenomglandmorphologyandthecorrelationvolume,optimizethespecificdrugmanufacturingline,accesstohigherbiologicalactivitythanpurespidervenomMethodsvenomglandanatomy,withMathematicalstatisticalanalysisofthevenomglandmorphologyandspiderbodymassrelationship,theuseofvenomglanddissectiondirectaccesstopropertiesofstableanduptotheamountofthespidertoxincrudetochickenredbloodcellsasamodeltestthevenompotency,andexplorethetoxinionchannelinhibitiontypeResultsvenomweightandbodymasswerepositivelycorrelated,inaccordancewiththeoptimizedpreparationandtechnicallines,obtainedmgArgiopespidervenom,ArgiopespidertoxintopromoteerythrocyteagglutinationandlysisroleinthecalciumionenvironmenttoxinonredbloodcellscleavagestrongerConclusionvenomdissectiontoobtainthemaximumamountofhigherpurityspiderpoison,whichinitiallyidentifiedthespidertoxinlectinandphospholipasewithtwocomponentsKeywords:Argiopespider,thevenomglandanatomy,venom,celltoxicityChinaAbstract:ObjectiveHavingadetailedmasteryabouttheArgiopeamoena‘smorphologicalcharacteristicsandlivinghabits,weanalyzedtherelevenceofpoisonglandweightandlengthtoitsbodyUsingtheoptimizedmethodweobtainedhigherbiologicalpropertiesandmorepurevenomMethodsWegottherelevenceofpoisonglandweightandlengthtoitsbodyusingstatisticalmethodbyanatomingpoisonglandIsolatingpoisonglandfromspider,obtainedstableandmaximumamountofvenom,wetestedtiterofvenomandexplorethetypeofinhibitionchannelstoxinusingchickenredbloodcellsasamodelResultsVenomgland‘sweightandbodymasswerepositivelycorrelatedWiththeoptimizedmeanpreparedmethod,fivemillingramsofcrudevenomweregotThevenomhaseffectofpromotingredbloodcellaggregatingandcrackingThebloodcellcrackedmoreseriouslyintheCalciumionsenvironmentConclusionMorequalifiedvenomcanbegotbyvenomglanddissectionWeinitiallyknowthatvenomcontainsphospholipaselectinandagglutininKeywords::ArgiopeamoenaAnatomyofvenomCrudetoxinCellphysiologySpidersareimportantnaturalenemiesofpests,paralysistoxintokillitspreySpidertoxincomplexcomponents,mainlyproteinsandmultiamines,proteinsincludingpeptideneurotoxins,cytotoxins,enzymes,kininanalogues,peptideaggregationandpeptides,andmanymoreamineswithATP,nucleotides,aminoacids,polyamines,amineandotherlowmolecularweightsubstancesandinorganicsaltsspidervenomwithavarietyofactivities,hasbeenfoundtocontainavarietyofspecificpeptideswithbody,canbeselectivelyandK,Na,Cachannelsubtypescombined~domesticequalityLiangandSongTigerhuwena,HainanandGuangxihuwenathetoxinsMacrotheleamoreindepthstudyArgiopespiderArgiopeamoenapartofAraneaeisArgiopegardenspiders,alargebody,forthespiderwebs,aredistributedinthesouthernprovincesofChina,manycomeandgoinsummerandautumntowintereggbagsArgiopespiderasmorepeopleliveinthemountainsoflushvegetationanddifficulttoraise,soonArgiopespidertoxinsinfewreportsapparatusandmaterialsInstrumentShanghaiPrecisionAelectronicbalance,OlympusCXmicroscopecamerasystem,ShanghaiXinmiaoDNPBSHeatedIncubatorsandDHGBSblastoventemperature,HunanStarScientificTGLBhighspeeddesktopcentrifuge,JiangsuKunshanKQDBCNCultrasoniccleaning,BeijingTechstereomicroscope,MitutoyoNKverniercaliper,micrometerShanghaiopticalmicroscopeandavacuumsuctionpumponthewingandYahyadryerandsoonArgiopespidercollectionArgiopespidercollectedfromtheforestcoldGuixiCityinJiangxiProvince,atotalof,themale,femaleArgiopespidermainlydistributedinthemountainoushighlands,clothonthesunnysiderotary,feedonlargerinsectsLord,joyinlivingareanetworkingaroundthebush,especiallycowbarnnexttomuchArgiopespiderfragile,withlonglimbs,easilyinjured,intheacquisitionprocessshouldbeextracareful,youcanusebutterflynetstospiderscapture,intothebox,putsomeleavesinordertomaintainhumidityandgoodventilation,avoidingoscillationsandexcessiveheatcollectioniscompletereturn,thespidershouldbeimplementedinatimelymanneronthesemishrubecosystemonthestockingResultsandMethodsArgiopespidervenomglandmorphologyandbodymasscorrelationArgiopespider’svenomglandislocatedinthesmallestchelatebasedfrontendtotheheadextended,andmoreintheheadandchestinfront,coveringtheganglionontheleftandrightinpairsandtendtothelongitudinalaxisofthespiderbodyInordertoquicklyandaccuratelyandefficientaccesstospidervenom,toavoidlossofmaterial,weArgiopespidervenomwereanatomy,aclearstructureandspecificlocationofthevenomgland,andthendescribethemorphologyofthevenomglandAnatomyoftheelectronicbalance,saidbeforetheweightofaspideroflargeindividuals,respectively,withaverniercalipermeasuredouthischestandabdomen,headlengthandwidth,whiletheindividualisasmallspiderwithamicroscopemicrometerheadandchestandabdomenmeasureoutthelengthandwidth,thenmeasurethelengthandremovethevenomglandwide,andweighedSpecificmeasurementresultsshowninTableand,regressionanalysisofthedatashowsthat:Argiopespiderinthesamesex,samehabitat,bodysizeisnotthesametime,thelocationandmorphologyofspidervenomandspiderpopulationdynamics,lifestylesandformshaveacertainvolumerelations,which,asthespider’svenomglandweightandtheirbodymasswaspositivelycorrelatedsignificantlyintothebodyofvenomglandlengthofitsheadandchestlengthbetweenthepositivecorrelation,significancetestwasextremelysignificantTableArgiopespideradultvenomglandweightassociatedwithindividualquality(omittedinTableArgiopespidervenomintothebodylengthandindividuallengthofdatadependenceanalysis(omittedArgiopespidervenomextractionandpreparationmethodsofoptimizationBecauseArgiopespiderwithlargetigerSPIDER,spiders,etcMaoYingZhaoJingcomparedtoadultsisrelativelysmall,theamountofnotmoretoxic,andtheappendageslong,difficulttoadoptpoison,wetrytoliveinthebodyinducedtotakedrugs,takedrugsandelectricalstimulationofanatomicalseparationofthevenomglandandothermethods,identifiedbythevenomglanddissection,afterrepeatedtestingcomparedtooptimizetheamountofadrugisnotsuitableforanatomicalspidervenomglandhomogenatestakedrugsmethod,followthesesteps:refrigeratorfirstspiderhalofreeze,accordingtoAolimbtodeterminetubelocationvenom,andthenfromthebackofthe“Dingtype”cutitscarapace,whichextendsupwarddirectionbythevenomglandfoundinthevenomglandtube,andcarefullyremovetheclamp,tolyophilizationWetookofdryvenom,intotimesthehomogenatemadeindeionizedwater,withrminrateofcentrifugemin,thesupernatantToaddanequalvolumeofsedimentsindeionizedwater,thencentrifugedsupernatantagainaddanequalvolumeofsedimentdeionizedwater,adjustingpHvalueto,homogenized,centrifugedandthesupernatantcombinedsupernatant,filteredtoremoveimpurities,obtainedsupernatant,andfrozen,andothersformedice,theicewillfreezethespiderpoisonplacedinavacuumdryer,vacuumdrying,thebottomofthedryerputtherightamountofcalciumoxideasadesiccant,andthenusethevacuumpump,pumpingprocess,toobserve,ifyoufindalotofspidertoxinsurfacebubbles,stopamomentapplyagainandagain,untilthebasicdry,intoacrystalparticlesofdifferentsizesuntilhofvacuumdryingisgenerallycompletedwithinathoroughlydrieddriedspiderServepowdereddrugorblock,scrapeddryDufendistributedintovials,dissolvedwaxseal,label,indicatingthepreparationofthespidertoxinDufendateandweight,theopaqueblackpaperbag,placedrefrigeratortosaveforyeardeteriorationinthewholeprocessoftakingdrugsandpreparation,youshouldfirstpayattentiontothewholeprocessmustbecleanandhealth,toensurethepurityofdrugstaken,followedbytheprocessoftakingdrugsasmuchaspossibleatlowtemperature,allreagentsmustbepreicebathArgiopespidervenomcompositionandtiterArgiopespidervenomcompositionandstructureobservationRoomtemperature,extractpreparationtoArgiopespidervenomformofpowder,venomwithanopticalmicroscope,andtimeswereunderthesheetorsheetvenomuniformcrystals,freeofotherparticulateimpuritiesFigureLinkstofreepaperdownloadArgiopespidervenomtestthecohesionandcrackingTakelofpreparedTaiheblackbonechickenredbloodcellsuspension(suspensioncelldensityofaboutmm,andlPBSbuffertomlofthePVCtube,thenaddlArgiopespidervenom,andOscillation,shakewellverticalconstanttemperatureatincubatoreverymin,withapipettegunlreactionsolutionobtainedinasingleconcaveslide,andwiththemicroscope,andphotographedgradientsbyrepeatedexperimentsTheresultsshowthatArgiopespidervenomsolutionontheTaiheblackbonechickenredbloodcellssignificantlypromotetheaggregationandcracking,whichcouldexplainthevenomcontainsaglycoproteinlectinandphospholipasecomponents,redbloodcellsandcanmediateadhesionandpromotecellcleavageFigureArgiopespidervenomcompositionandpromoteexplorationofionchannelsGoodpreparationinl,respectively,Taiheblackbonechickenredbloodcellsuspension(suspensioncelldensityofaboutmmtoaddthecorrespondingmolLKCl,andmolLCaClsolution,deployedtoml,dividedintothecontrolgroup,theformertwowereaddedtolArgiopespidervenomandlgardenspidervenombigbelly,anotheronebyaddinglPBSbufferlocatedmin,min,min,minandmindifferentreactiontime,subboardtotakefluidinthebloodcounts,usingmicroscopeandcountedspecificreactionresultsshowninFigure~Throughtheanalysisofthecomparisonchart,youcanlearn:Argiopegardenspiderspidertoxinsandbigbellytoxinsaretopromotetheroleofredbloodcellhemolysis,andthebigbellygardenspidertoxinbecomesmoreapparentMoreover,thetoxiceffectonredbloodcelllysisinthecalciumionenvironmentmorethanthepotassiumionenvironmentspeculatethereasonmaybetwovenomligandincalciumchannelactivationgatecomponents,resultinginrapidcalciumionsintoredbloodcells,acceleratedhemolysis,alsomayexistinspidervenomphospholipasecleavageofredbloodcellmembraneisoneofthereasonsDiscussionAtpresent,thespidercollectionofmostlydruginducedmethodandtheelectricshockmethod,asArgiopespiderindividualsisrelativelysmall,thinlimbclawsshort,livetakendrugsextremelydifficultInthispaper,thelifeofArgiopespiderbehaviorandmorphologyofapreliminarystudybasedontheoptimizationofthevenomglanddissectionwithimprovedhomogenization,werepreparedsuccessfullyextractedwithstablepropertiesandphysiologicalactivityofthevenommg,forthepurificationoftoxinsandstudyfollowupbasisfollowupexperimentsshowedthattheuseofvenomglandhomogenateanatomymethodtoobtainthemaximumamountofhigherpurityspiderpoisonintherealmicroscope,withanatomicalcuttheclawscutoutlimbsandcarapacepoisonsac,suchaspoisonsacpuncturedwithneedles,withouthomogenization,toavoidfurthercontaminationofnontoxinproteinsdeionizedwatertowashthevenomout,rminmincentrifugationsupernatant,freezedrying,toobtainthetoxincanbeobservedunderamicroscope,uniformcrystalsbythespidertoxinonchickenbloodtoxicologicalresponsesofredbloodcells,whichinitiallyidentifiedthespidertoxinlectinandcontainstwocomponentsphospholipaseInaddition,calciumisalsofoundintheenvironmentofspidertoxinonhemolysisandagglutinationofredbloodcellsismoreintense,thereasonSpidervenommaycontainasubstanceactingontheplasmamembranecalciumchannelliganddoor,thuscontributingtocalciumionsintoredbloodcellsleadingtocellswellingfasthemolysis,perhapsthevenomofthespiderisatypelectincadherinincalciumionactivatedredbloodcellscaninduceadhesionoraggregation,reduceOCOexchangerate,respiratorydistressandevendeathcausedbythebodyReferencesYuanfengHui,BaiTao,CaoBo,etalThemedicinalvalueofspiderOverviewJMATERIAMEDICA,,():FeiRui,YangYangSpiderToxinprofileandapplicationJJilinUniversity,,():WangJianhui,GuVinaSpiderPeptideToxinsResearchJSpidershaped,,():LawofHair,YanRupertSpidervenomglandanatomyofresearchJLaserBiologySinica,,():HuangRenhuai,SongPingLiangSpiderPeptideneurotoxinsNewDevelopmentJLifeSciences,,():MalcolmRBrown,DavidDSheumackAminoacidsequenceofversutoxin,alethalneurotoxinfromthevenomofthefunnelwebspideratraxversutusJBiochemJ,:Gray,MRSutherland,SKArthropodvenomsSpringerVerlag,Berlin:ppAndrJunqueiraZaharenko,WilsonAlvesFerreiraProteomicsoftheneurotoxicfractionfromtheseaanemoneBunodosomacangicumvenom:NovelpeptidesbelongingtonewclassesoftoxinsJoacirStolarzOliveira,MichaelRichardson,,(:KristianStrmgaard,IanMellorAMPAreceptorligands:SyntheticandpharmacologicalstudiesofpolyaminesandpolyaminetoxinsJMedicinalResearchReviewsMarch,(:GlennFKing,MargaretCGentz,PierreEscoubas,etalArationalnomenclaturefornamingpeptidetoxinsfromspidersandothervenomousanimalsJToxicon,,():HelmutKubista,RobertaAMafra,YoumieChongCSTX,atoxinfromthevenomofthehuntingspiderCupienniussalei,isaselectiveblockerofLtypecalciumchannelsinmammalianneuronsJNeuropharmacology,,():SongPingLiangPoisonousidentificationofthebiologicalactivityJZoologicalResearch,,():ZhuoChunhuiArgiopespiderArgiopeamoenaIfobservationandecologicalcharacteristicsofthespiderArgiopespiderwebstructure,mechanicalpropertiesofsilkDragglingDMasterThesis:LifeSciences,SichuanUniversity,Linkstofreepaperdownload

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Argiope spider venom extraction and preparation

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