PROTEIN EXPRESSION PROFILES FOR PWS DISEASE

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 (1)假设：异常血管再生被假设为主要的 激光治疗后PWS复发的原因。我们 假设血管生成途径在此过程中起重要作用 血管再生，因此抑制血管生成途径可能导致 改善预后。 (2)实验设计：将使用小鼠或小鼠动物模型进行 这个项目动物皮将被激光照射。皮肤组织 在激光治疗之前和之后将被收集、固定和切片。 一些关键的血管生成途径分子，包括VEGF、mTOR、Akt、PI3K， 采用荧光免疫组化法检测MAPK等。 (3)在我们建立了一个坚实的数据，从动物模型，我们将执行 在人体皮肤上进行类似的实验。简言之，正常和PWS人皮肤将 由纳尔逊医生在患者同意的情况下收集。皮肤组织 （约1mmx1mm）立即固定过夜并切片。 参与血管生成的关键分子将通过以下方式进行研究： 荧光免疫组织化学 (4)将使用两种荧光团：488和549。单染色或双染色 将同时进行。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. (1) Hypothesis: Abnormal vessels regrowth is hypothesized as the main reason of recurrence of PWS after laser treatment in patients. We hypothesize angiogenesis pathway plays an important role during this vessel regrowth, thus inhibition of angiogenesis pathway may result in an improved prognosis. (2) Experimental design: Hamster or mouse animal model will be used for this project. The animal skins will be radiated by laser. The skin tissues before and after laser treatment will be collected, fixed, and sectioned. Some key angiogenesis pathway molecules, including VEGF, mTOR, Akt, PI3K, MAPK etc will be investigated by fluorescent immunohistochemistry. (3) After we establish a solid data from animal model, we will perform similar experiments in human skin. Briefly, normal and PWS human skin will be collected by Dr. Nelson with patients' consents. The skin tissues (around 1mmx1mm) will be fixed immediately for overnight and sectioned. The key molecules involving in angiogenesis will be investigated by fluorescent immunohistochemistry. (4) Two fluorophores will be used: 488 and 549. Single or double staining will be performed simultaneously.