ydevaporfor5miairdriedwith40℃for30miwashedwithtfor15min×3.thenthegridswereusedtoinvestigatehyaluronanbyelectrogoldmicroscop:sixnickelgridswereincubatedwithha-biotin,theothersixwereincubatedwithtascontrol.
resultinthecontrol,theperitonealmesotheliumwascoveredwithmicrovilliwhereasnosurfacelayerwasoerved.however,inthegaggroup,thereaearedtobeadiscontinuousamorphouslayercoveringthemesothelialcells.intheph1group,acontinuousamorphouslayercoveredtheperitonealsurfaceanditsthicknedependedontheabundanceofmicrovilli.thislayerwasmuchbetterpreservedintheph2groupwiththickneupto10μm.inthetaicacidgroup,notonlythisstagnantlayercouldbeeasilyseen,wecouldbutalsooervetherewerealotoflamellarbodiesinthelayerandiidethemesothelialcells.theimprintshowedperitonealsurfacelayerwithlatticesstructure.hapositivesignalcouldbeoervedintheexperimentalgroup,whereasinthecontrolgroupnogoldenparticlewasoerved.
3.hyaluronansynthase(has)2ecificantiseeoligonucleotidesinhibittheformationofpericellularcoatinhumanperitonealmesothelialcells(hpmcs)
asaprerequisitestudy,theexpreionofeachhasmrnainculturedhpmcswasmeasuredbyreversetracription-polymerasechainreaction(rt-pcr).onlyhas-2andhas-3mrnaexpreionweredetected,thelevelofhas-2mrnaexpreionwasabout10foldhigherthanthatofhas-3.has-2ecificantiseeoligonucleotideswerethentrafectedintoculturedhpmcs.after0,8,48and24hoursthehpmcswereoervedusingparticleexclusionaayandtheirhas-2mrnaexpreionwasdetectedagain.theresultsshowedthathas-2mrnaexpreionwasmostlyinhibitedatthe24hourasmea
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