简介:摘要目的构建并包装大鼠Six2基因过表达慢病毒及建立稳定过表达Six2基因的MES23.5细胞株。方法采用RT-PCR方法扩增Six2基因片段并将其连接于含有嘌呤霉素抗性的pLV-CS2.0-N-HA载体质粒,采用慢病毒包装四质粒系统(pRRE/pVSVG/pREV/pLV),用转染试剂Lipofectamine2000将四质粒按一定比例转染293T细胞;收集慢病毒上清并感染MES23.5细胞株,嘌呤霉素筛选稳定表达Six2的MES23.5细胞株,Westernblotting检测Six2蛋白的表达。结果限制性内切酶酶切结果表明在900bp处有一明显条带;Six2测序结果显示序列与genebank上的序列完全一致;嘌呤霉素筛选病毒感染的MES23.5细胞,传代筛选第5代后细胞几乎无死亡现象;Westernblotting结果显示,在分子量为23.4kDa处有相应条带。结论成功构建过表达Six2基因的慢病毒表达载体;得到了较高滴度的病毒悬液;建成稳定表达Six2基因的MES23.5细胞株,此研究为进一步探讨Six2基因的功能提供了良好的研究工具。
简介:TheRatnagotravibbāga(abbr.RGV)isoneofthemostinfluentialtextsamongIndicworksthatteachtheBuddha-naturedoctrineinTibet.NotranslationofthetextseemstohaveexistedinTibetbeforetheⅡthcentury,inasmuchasnocatalogueoftheimperialperiod(the9thcentury),suchasthelDandkarmaandvPhangthangma,showsanyrecord.AlthoughonlyasingleTibetantranslationisextant(thatofrNgogBloldanshesrab[1069-Ⅱ09]andSajjana),vGosLotsābagZhonnudpal(1392-1481)reportsthatuptohistimesixtranslationshadalreadybeenmade:1
简介:Inthepresentpaper,theauthorputsforwardsixkeypointsforacupuncturetreatmentofdiseases,namely,①carefulexamination,②definitediagnosis,③preciseandappropriateidentificationofsyndromes,④accuratelocationoftheacupoint,⑤flexibleapplicationofneedlingmanipulations,and⑥“Deqi”.Thefirstthreeaspactsarethefoundation,accuratelocationandflexibleneedlingmanipulationsarealsotheprerequisiteforeffectivetreatmentofdiseases.Inaddition,soundtheoreticalbasicknowledgeofbothtraditionalChinesemedicine(TOM)andmodemmedicine,andflexiblyapplyingsuitableneedlingmaneuvers,stimulatingquantityanddurationofneedleretaininginaccordancewiththeconcretestateofdiseaseandthepatient'sconditionsarealsoveryimportantinclinicalpracticeofacupuncture.
简介:ThesixcuneiformtextsbelowarehousedbytwoJapaneseuniversitiesandrecentlyattractedmyattention.WiththekindpermissionsofProf.KoichiYoshiikeoftheFacultyofForeignStudies,theAichiPrefecturalUniversity,andofProf.RyozoMatsumoto,ExecutiveDirectoroftheTokaiUniversityLibrary,theyarepublished.
简介:ⅠStomachacheStomachacheisacommonsymptomseeninacuteandchronicgastritis,pepticulcer,andgastricneurosis,etc..Thepainappearssuddenlyorslowly,anditsometimesreferstothehypochondriumortheback.Insomecas-es,theremaybenausea,vomiting,poorap-petite,acidregurgitationorevenhematemesisandtarrystool.IntheviewpointoftraditionalChinesemedicine,stomachacheismainlycausedbyinvasionofcold,irregularfoodin-takeandemotionalfactors.