简介：在大多数谷物庄稼，phytic酸是磷的主要存储形式，它能减少磷酸盐的简历可获得性。phytase的转基因的表示被认为是在转基因的植物免除磷酸盐phytate的一个有效方法。在这研究，工厂表示向量，包含重组体phytase基因由玉米ubiquitin(Ubi)开车倡导者经由Agrobacterium-mediatedtransformation被构造并且介绍进一个精英米饭变化。在实验期间，15根独立转基因的米饭线的一个总数被改革。PCR和南部的污点的结果显示目标基因集成于转基因的米饭工厂的染色体。而且，提取fromtheimmature几根转基因的线播种的全部的RNA的RT-PCR分析证明重组体phytase基因能通常被表示。无机的磷内容，两个都比在untransformed野生型在转基因的工厂在成熟种子和叶是显著地更高的。

简介：Aim:Tostudytheeffectofprothymosinα(ProTα)asafusionproteinonthesecetionofINF-γ,IFN-αandTNF-αinvitro.Methods:Theinvitrostudywascarriedoutontheculturedofsplenocytes,splenicandperitonealmacrophagesisolatedfromBalb/cmice.SplenocyteswereincubatedwithvariousconcentrationsofProTα(1×10^-7-1×10^-10mol·L^-1)withorwithoutConA(5μg·mL^-1)for72h.SplenicandperitoealmacrophageswererespectivelytreatedwithProTα(1×10^-7-1×10^-10mol.L^-1)inthepresenceofLPS(10μg·mL^-1)for24h.ThenINF-γ′s,INF-α′sandTNF-α′slevelsinthesupernatantweredetecedbyELISA,Results:ProTα(1×10^-7)mol.L^-1)wasfoundtoobviouslyincreaseINF-γlevel(P<0.05)inthesupernatantofsplenocytescomparedwiththecontrolgroup.Moreover,ProTα(1×10^-7mol.L^-1)significantlyinducedthesecretionofINF-α(P<0.01)andTNF-α(P<0.01)insplenicandperitonealmacrophages.Conchusion:Invitro,ProTαcouldincreasethesecretionofIFN-γ,IFN-αandTNF-α.

简介：以便学习TGF-beta1的结构功能细节，重组体老鼠TGF-beta1的成熟形式在细菌被表示。TGF-beta1(氨基酸279-390)的112氨基酸的carboxyl终端部分的合成被一个可诱导的基因表示系统基于抗菌素T7RNA控制。这个系统允许重组体TGF-beta1的活跃、选择的合成。在减少条件下面在SDS-polyacrylamide胶化上决定的表示TGF-alpha1单体的分子量是大约13kD。连续净化洗与纯化步是足够的净化表示产品到同质的单个胶化过滤结合了。氨基终端的定序表明重组体蛋白质的N终端与出版数据相同。在西方的污点分析，重组体多肽对polyclonalTGF-beta1抗体显示出优秀antigenicity。在这研究表示的成熟重组体老鼠TGF-beta1提供一个有用工具因为未来详细说明了结构、功能的研究。

简介：Prolactinisamultifunctionalhormonethatexertsmanyseparatefunctionsandactsasanimportantconnectionbetweentheendocrineandimmunesystems.Thereareincreasingresearchesimplicatingtheroleofprolactininhematopoiesis.Enhancederythropoiesisinpregnantwomenanddirecterythropoieticeffectsinvitroofplasmaeitherfrompregnantorlactatingmicehavebeenreported.Furthermore,regressionoferythroblasticleukemiahasbeenobservedinasignificantnumberofratsafterhypophysectomy.Inthisstudy,theeffectsofrecombinanthumanprolactin(rhPRL)onhematopoiesiswereassessedinirradiatedmice.MiceweretreatedwithrhPRLforfiveconsecutivedaysafterexposuretoalethaldoseorasub-doseirradiation.Prolongedsurvivalrateandincreasederythropoiesiswereobservedintheirradiation-inducedmyelosuppressivemice.ItwasconcludedthatrhPRLmightactonerythropoiesisandcouldbeapotentialcandidateforthetreatmentofirradiation-inducedmyelosuppresioninclinic.Cellular&MolecularImmunology.

简介：AgreatamountoffoodbornepathogenswereGram-positive(G?)bacteria,athreattopublichealth.Inthisstudy,consideringthebindingabilityofnisintowardsG-bacteriaandthestablefluorescentabilityofEGFPprotein,afluorescentnisin–EGFPproteinprobewasconstructedbyageneengineeringmethod.NisinandEGFPwereusedasthereceptorandfluorophore,respectively,todetectG?bacteria.ThenisinandegfpgenewereamplifiedseparatelyaccordingtothesequencepublishedinGenBankusinguniqueprimers.ThetwogeneswereclonedintoapET-28b(?)vectorresultinginapET-28b(?)–nisin–egfpvector.ThevectorwastransferredintoEscherichiacoli(E.coli)BL21(DE3)forexpression.Theexpressedproteinwasextracted,purifiedbyaNi–NTAcolumn,andthentestedbytheSDS-PAGEmethodtoconfirmitsmolecularweight.Listeriamonocytogenes(L.monocytogenes),Staphylococcusaureus(S.aureus),andMicrococcusluteus(M.luteus)wereusedastherepresentationsofG?bacteria.E.coliO157,representingthegram-negative(G-)bacteria,wasusedasanegativecontrol.Thebindingspecificityoftherecombinantproteinwasperformedontwotypesofbacteriaandthendetectedthroughfluorescentmicroscopy.Theresultsindicatedthatthenisin–EGFPprobecoulddetectG?bacteriaat108CFU/mL.

简介：可移植的试验性的肿瘤模型被构造对肿瘤复发和转移学习recombinant人interleukin-15(rhIL-15)的活动。结果证明那肿瘤小瘤形成被延迟，肿瘤生长与rhIL-15在处理以后在LA795肺腺癌的下的肿瘤模型被禁止，并且与rhIL-15对待的T739忍受肿瘤的老鼠的幸存率比与盐的任何一个或与rhIL-2的一样的剂量对待的老鼠的高得多。这indicats那rhIL-15antitumor最好在一样的剂量水平比rhIL-2完成。在一些，rhIL-15对待老鼠，皮下地接种的肿瘤房间被根除，没有肿瘤形成甚至在肿瘤以后的138天房间接种。没有肿瘤的老鼠与实时肿瘤房间被重新质问，没有肿瘤这些老鼠在所有在下列二个月内重新发生，显示全身的免疫开发了的那长持续的antitumor。肿瘤复发和转移与rhIL-15，然而并非与rhIL-2的一样的剂量在处理以后显著地被禁止，这也被显示出，在LA795肺腺癌的皮下地并且静脉内地传播的肿瘤模型。同时，splenocytes的CTL和NK房间活动从与任何一个rhIL-15被对待的忍受肿瘤的老鼠获得了或rhIL-2是显著地提高的两个。然而，CTL和NK房间活动的改进在rhIL-15是更重要的在rhIL-2比那对待老鼠对待的老鼠。这建议在vivo的rhIL-15的反肿瘤效果被在肿瘤免疫者反应提高CTL和NK房间活动完成。细胞与分子的免疫学。2008；5(3):189-196。

简介：Theenhancedcardiaccontractilityeffectofhumanrecombinantgrowthhormone(hr-GH)onthecongestiveheartfailure(CHF)wasstudiedonthepig.Tobuildapigmodelofcongestiveheartfailure,atemporaryartificialcardiacpacemakerwasimplantedinthepig'sbodyandpacedat220beatsto240beatsperminutefor1week.Afterthemodelofcongestiveheartfailurewassuccessfullysetup,thefrequencyofthepacemakerwaschangedto150beatsto180beatsperminutetomaintaintheCHFmodelstable.Pigsweredividedintothreegroups:Thehr-GHgroupinwhich0.5mg/kgperdayofhr-GHwasadministratedintramuscularlyfor15days,theinjectioncontrolgroupinwhichanequalamountofphysiologicalsalinewasinjectedintramuscularly,andanormalcontrolgroup.Theleftventriculardiastolicendpressurewas(10.60±2.41)mmHginthehr-GHgroup,but(19.00±3.81)mmHginthesalinecontrolgroup(P＜0.01);Cardiacoutputwas(1.86±0.13)L/mininthehr-GHgroup,but(1.56±0.18)L/mininthesalinecontrolgroup(P＜0.05);Peripheralmin)-1inthesalinecontrolgroup(P＜0.05);±dp/dtmaxwas(2900±316.23)and(2280±286.36)inthehr-HGgroupandthesalinecontrolgrouprespectively(P＜0.05).Theresultsshowthathr-GHenhancesmyocardialcontractilityofCHF,andtheCHFmodelbuiltbyatemporaryartificialcardiacpacemakeratahighrateofstimulationisreasonableandapplicable.

简介：Objective:Inthisstudy,weexaminetheeffectsofrecombinantadenovirus-p53(rAd-p53)onthepancreaticcarcinomacelllineSW1990.Specifically,wedetermineifexpressionofrAd-p53sensitizesthesecellstoradiation.Methods:FollowingtransfectionofSW1990cellswithrAd-p53,wemeasuredexpressionofP53,P21andBaxbyimmunocytochemistry.Bothtransfectedandcontrolcelllineswereirradiatedwitharangeofdoses,andthesurvivalfractions(SF)werecalculated.Dosesurvivalcurveswereconstructedandmodeledforcomparison.Results:TransfectionofSW1990cellswithrAd-p53resultedinincreasedexpressionofP53,P21andBaxinatime-dependentmanner.At96haftertransfection,89.92%ofcellsexpressedP53,56.8%expressedP21,and76.50%expressedBax.TheSFfollowingradiationwaslowerintherAd-p53transfectedcellscomparedtothecontrolcells,suggestingthatrAd-p53sensitizesSW1990cellstoradiation(D0fortheexperimentalandcontrolgroupswas2.199and2.462,respectively).Conclusions:UseoftheadenoviralvectorisaneffectivemeansoftransfectingSW1990cellswithwild-typeP53,andthissensitizesthecelllinetoirradiation.ThisworksuggeststhatcombiningrAd-p53withradiationtherapyinpancreaticcancermaybetherapeuticallybeneficial.

简介：BACKGROUND:Erythropoietinandrecombinanthumanerythropoietin(rhEPO)inhibitapoptosisofmotorneuronscausedbyspinalcordinjuryandbraindamageinrats.However,itstillremainstobeshownwhetherrhEPOcanprotectfacialmotoneurons(FMNs)aswell.OBJECTIVE:TotesttheneuroprotectiveeffectsofrhEPOoninjuredFMNs,aswellastheinfluenceonCaspase-3expression.DESIGN,TIMEANDSETTING:Randomized,controlled,animalexperiment.ThisstudywasperformedattheCentralLaboratoryofBasicMedicalCollege,ChongqingMedicalUniversityfromJanuarytoOctober2007.MATERIALS:Seventy-fivefemaleSDrats,weighing210–230g.rhEPOinjectionwasprovidedbySanshengpharmaceuticalscompany,ShenyangCity,LiaoningProvince,China,andtheLicensenumberwasHMLNS20010001.METHODS:Atotalof75femaleratswererandomlydividedintorhEPOtreatment,control,andshamoperationgroups,with25ratsineachgroup.RatmodelsoffacialnerveinjurywereestablishedintherhEPOtreatmentgroupandthecontrolgroupbycrushingthemaintrunkoftheleftfacialnerve.Surgicalmicroscopicobservationofthefacialnervedamagedisplayedperineurialdisruption.Theleftstylomastoidforamenoftheshamoperationgroupwereonlyexposed,butwithoutnerveinjury.TherhEPOtreatmentgroupwastreatedwithrhEPO(5000U/kg,i.p.)oncefollowinginjuryandonceadayfortwoweeks.Thecontrolandshamoperationgroupsweretreatedwiththesamedoseofnormalsaline(i.p.),oncefollowinginjuryandonceadayfortwoweeks.MAINOUTCOMEMEASURES:Ratsweresacrificed3,7,14,21,and28daysafterinjury,FMNsurvivalafterfacialnerveinjurywasanalyzedbyToluidinebluestaining,andthensurvivalratios(L/R)werecalculated.ThenumberofapoptoticprofilesintheinjuredFMNswereevaluatedbyTUNELstaining.ExpressionofCaspase-3inthefacialnucleuswasdetectedbyimmunohistochemistrymethods.RESULTS:Atotalof75ratswereincludedinthefinalanalysis.FMNsurvivalratios,bothin

简介：Objective:Toassessthesafetyandclinicalantiangiogeniceffectofrecombinantadenovirus-p53(rAd-p53)combinedwithhyperthermiaplusornotplusradiotherapyinadvancedcancer.Methods:ExpressionofVascularepithelialgrowthfactor(VEGF)afterintratumoralinjectionofrAd-p53wasassayedbyimmunohistochemistry(IHC)imaging.Forty-fourpatientswithadvancedcancerwereenrolledintothisclinicalstudy.ThepatientswereintratumorallyinjectedwithrAd-p53(Gendicine)atadoseof1×1012vponceaweek,withatotalof4-54(mean7.7)times.Totalof4-29(mean8.5)timesofhyperthermiawasgiventothepatients.Amongthe44patients,30patientswereconcurrentlyaddedwithradiotherapyofatotaldose30-76Gy/15-38f/3-8w(mean58Gy).Results:BeforeandafterintratumoralinjectionofrAd-p53,theVEGFIHCpositivecellscoreswere2.80and1.50,respectively(P=0.031).ThetreatmentofrAd-p53combinedwithhyperthermiaplusornotplusradiotherapyinadvancedcancerachievedCRrateof13.60%(6/44),andPRrateof29.6%(13/44),andthustheeffectiveratewas43.2%.Inadditionto6patientswithCR,19patients(19/38,50.0%)hadlowdensityarea(LDA)ofmorethan50%areaonCTimagewithintumorindicatingtumortissuenecrosis.Conclusions:OurdataindicatethatrAd-p53inhibitsVEGFexpressionandangiogenesis,andpromotestumornecrosisandshrinkageinducedbyhyperthermiaplusornotplusradiotherapyinadvancedcancer.

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简介：Objective:TheobjectiveofthisstudywastoexaminetheeffectofhypothyroidismonhearingfunctioninpatientssurgicallytreatedfordifferentiatedthyroidcancerandsubsequentlyexperiencedhypothyroidismduringpreparationforfollowupI-131scintigraphyscanbyeitherrecombinanthumanthyroidstimulatinghormone(rhTSH)treatmentorthyroidhormonewithdrawal(THW).Methods:Atotalof55patientsundergoingI-131scintigraphyscanfollowingsurgeriesfordifferentiatedthyroidcancerwereincludedinthestudy,including25patientspreparedbyadministrationofrecombinantTSH(rhTSHGroup)and30patientsbythyroidhormonewithdrawal(THWGroup).Results:Airconductionthresholdsat1,2and4kHzforbothearswerehigherduringhypothyroidperiodthanduringeuthyroidperiodforpatientsintheTHWgroup(p<0.05)butnotforpatientsintherhTSHgroup.Conclusion:Sensorineuralhearinglosswasdetected,especiallyatlowfrequencies,inpatientswithDTCaftersurgicaltreatmentwhosehormonereplacementtherapywaswithdrawnbutnotinthosereceivingrhTSH.ItisthereforepreferredtouserhTSHwhenpreparingforI-131scintigraphyscaninpatientsatriskforhearingloss.

简介：由202根线组成的一张回交重组体近交系人口从XieqingzaoB//XieqingzaoB/Dongxiang野米饭被开发。人口在植物hopper抵抗和收益特点上是assayedwithDNA标记和phenotyped。A连锁图在于of119脱氧核糖核酸标记并且跨越了因为在12个米饭染色体上的1188厘米被构造。Thirty-twochromosomal片断替换线在标记loci基于XieqingzaoB等位基因的百分比被选择。这些线具有为基因印射和相异的基因基因渗入的大潜力。

简介：Inthepresentstudy,weconstructedalentivirus,FIV-CMV-GFP-miR-7-3,containingthemicroRNA-7-3geneandthegreenfluorescentproteingene,andusedittotransfecthumangliomaU251cells.Fluorescencemicroscopyshowedthat80%ofU251cellsexpressedgreenfluorescence.Real-timereversetranscriptionPCRshowedthatmicroRNA-7-3RNAexpressioninU251cellswassignificantlyincreased.ProliferationwasslowedintransfectedU251cells,andmostcellswereintheG1phaseofthecellcycle.Inaddition,theexpressionoftheserine/threonineproteinkinase2wasdecreased.ResultssuggestedthattransfectionwithalentiviruscarryingmicroRNA-7-3caneffectivelysuppressepidermalgrowthfactorreceptorpathwayactivityinU251cells,arrestcellcycletransitionfromG1phasetoSphaseandinhibitgliomacellgrowth.

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简介：Objective:ToprovideahighlyefficientadenoviralvectorAd-CMV-hTGFβ1forthestudyofgenetherapyforreversionoftheintervertebraldiscdegeneration.Methods:Anewlydevelopedrecombinantadenoviralvectorconstructionsystemwasusedinthestudy.ThecDNAofhTGFβ1wasfirstsubclonedintoashuttleplasmidpShuttle-CMV.TheresultantplasmidwaslinearizedbydigestingwithrestrictionendonucleasePmeI,andsubsequentlytransformedintoE.coll.BJ5183cellswithanadenoviralbackboneplasmidpAdEasy-1.Recombinantswereselectedbykanamycinresistanceandconfirmedbyrestrictionendonucleaseanalysis.Finally,therecombinantplasmidlinearizedbyPmeIwastransfectedinto293cells.Recombinantadenovirusesweregeneratedwithin2weeks.Results:TherecombinantadenoviralplasmidswerecutbyBamHIandPacIrespectively,andthediagnosticfragmentsappearedin0.8%agaroseelectrophoresis.Theinfected293cellsshowedevidentcytopathlceffect(CPE).TheproductionsofPCRconfirmedthepresenceofrecombinantadenovirus.TheexpressionofhTGFβ1wasverifiedbyimmunohistochemicalstaining.Conclusions:ThesuccessfulgenerationoftheadenoviralvectorAd-CMV-hTGFβ1andtheconfirmationoftheinterestgeneexpressionmakeitpossiblefortheexperimentalstudyofthereversionoftheintervertebraldiscdegenerationbygenetherapy.

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简介：AIMMacrophagemigrationinhibitoryfactor(MIF)isapro-inflammatorycytokineinvolvedinthepathogenesisofavarietyofautoimmuneandinflammatorydiseases,includingarthritis,glomerulonephritis,Gram-positiveandGram-negativesepsis,andatherogenesis.RecentstudiesshowedthatCD74(antigen-associatedinvariantchainⅡ)isahigh-affinitymembrane-bindingproteinforMIF.ThepurposeofthepresentstudywastoexpresstherecombinanthumanCD74inE.coliandinhibittheactivityofMIFbyusingrecombinantCD74invitro.

简介：Objective:ThispaperistoexploreamethodoftransferringhumanSDF-1anditsmutantSDF-1/54intrakinegeneintoCOS-7cellsfordeterminingtheirexpressionandsubcelluarlocalizationofthefusionprotein.Thiscouldofferfeasibilityforinhibitingthemetastasisofmalignanttumorsbyphonotypicknockoutforblockingfunctionalexpressionofreceptoronthecell-surface.Methods:AmplifythetargetgenewithPCRfromtheconstructedplasimidSDF-WT-Gly×4-Dec/PET-30a(+)withaC-terminalretentionsignalfragmentKDEL.AfterthepcDNA3.1/SDF-1/KDEL,pcDNA3.1/SDF-1/54/KDEL,pEGFP/SDF-1/KDELandpEGFP/SDF-1/54/KDELeukaryoticexpressionvectorswereconstructedandtheDNAsequencewasaccurate,theyweretransferredintoCOS-7cellswithliposome.Theexogenousexpressionswereobserved,fusionproteinSDF-1/HisandSDF-1/54/HiswereconfirmedbyWesternblot,andtheSDF-1/EGFPandSDF-1/54/EGFPweredeterminedbyLaserScanningConfocalMicroscopy.Results:Fourexpressionvectorswereconstructedsuccessfully,thefusionproteinSDF-1/KDEL/HisandSDF-1/54KDEL/HisexpressedinCOS-7cells.SubcelluarlocalizationanalysisshowedthatSDF-1/KDEL/EGFPandSDF-1/54/KDEL/EGFPwerelocatedmainlyinendoplasmicreticulum.Conclusion:FourexpressionvectorspcDNA3.1/SDF-1/KDEL,pcDNA3.1/SDF-1/54/KDEL,pEGFP/SDF-1/KDELandpEGFP/SDF-1/54/KDELwereconstructedsuccessfully,whichcouldexpressineukaryoticcellandlocatemainlyintheendoplasmicreticulum.