PROTEIN EXPRESSION PROFILES FOR PWS DISEASE

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 (1)假设：以异常血管再生为主要假说 激光治疗后PWS复发原因分析。我们 假想的血管生成途径在此过程中起着重要作用。 血管再生，因此抑制血管生成途径可能导致 改善了预后。 (2)实验设计：实验动物模型为仓鼠或小鼠。 这个项目。动物的皮肤将受到激光的辐射。皮肤组织 对激光治疗前后进行收集、固定、切片。 一些关键的血管生成途径分子，包括血管内皮生长因子、mTOR、Akt、PI3K、 MAPK等将用荧光免疫组织化学方法检测。 (3)在建立坚实的动物模型数据后，我们将进行 在人类皮肤上进行了类似的实验。简而言之，正常和PWS人类皮肤将 由纳尔逊医生在征得病人同意的情况下收集。皮肤组织 (约1 mm×1 mm)将立即固定过夜并分段。 参与血管生成的关键分子将通过 荧光免疫组织化学。 (4)将使用两个荧光团：488和549。单染或双染 将同时执行。

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.