DEVELOPMENT OF CLINICAL MARKER FOR PERILYMPH FISTULA

围绕外淋巴瘘（PLF）诊断的问题存在许多 对耳科医生的挑战 由于确定的标准， PLF的术前诊断和术中确认 虽然已经建立，但病例定义标准不确定。 之前 临床研究提示，PLF可能比 以前认识到，但其特点是缺乏前瞻性 设计和适当的控制组。 这项研究的第一阶段建议利用 外淋巴蛋白，以开发临床上适用的标记物， 对PLF的术中诊断具有特异性，并且足够敏感 检测低至2-5微升量的外淋巴， 被其他外来液体污染。 还计划开展II期临床试验： 1)假设开发了PLF的敏感标记物， 一项机构前瞻性观察性研究预计将研究 可证实PLF患病率和确定的诊断预测 听力和前庭功能障碍患者的标准。 2)解决潜在假阴性结果的不确定性 间歇性PLF和微瘘，一项前瞻性随机 涉及前庭功能障碍患者的临床试验是 拟比较手术治疗假定PLF的疗效 对照组采用前庭康复治疗。 本研究的目的是了解发展中国家的 视网膜细胞实现其特化的机制 子类型 玻璃突变特异性地去除了视网膜的感光神经元， 果蝇的视觉系统;这些细胞开始发育为神经元， 不能表达感光细胞特异性基因，随后死亡。玻璃 编码具有五个锌指结构域的蛋白质;这种蛋白质已经被 在其他生物体中作为转录因子发挥作用。 因此玻璃 可能直接调控感光细胞特异性基因 表情 我们已经证明，玻璃蛋白在体外结合到 来自一个这样的基因（视紫红质）的增强子元件的序列。玻璃 在发育中的视网膜的所有细胞中都有表达，但它只是 在发育中的光感受器中具有活性（通过显示对 玻璃DNA结合位点的报告基因）。 因此，玻璃的调节 活性对于感光细胞的发育至关重要， 在两个层面上：在转录，并在翻译后（通过传入 位置信号）。 为了实现这一双重监管，我们将进行两次 项目： 1)为了了解glass的转录调控，我们将进行一项 通过突变研究对glass基因启动子进行功能分析。 2)为了鉴定与玻璃相互作用的新基因，我们将使用一系列 第一个将是显性增强子， 弱玻璃区的抑制剂。 初步筛选已经成功 测试了这种方法的可行性（我们已经恢复了三个这样的 突变）。 一旦被隔离，我们将描述我们的小说的效果 从长远来看，克隆基因， 它们的分子功能

Issues surrounding the diagnosis of perilymph fistula (PLF) present many challenges to the otologist. Since definitive criteria for the preoperative diagnosis and intraoperative confirmation of PLF have not been established, case definition criteria are uncertain. Prior clinical studies raise the suggestion that PLF may be more common than previously appreciated, but are characterized by a lack of prospective design and appropriate control groups. Phase I of this study proposes to utilize the unique constituency of perilymph proteins to develop a clinically applicable marker that is specific for the intraoperative diagnosis of PLF, and sensitive enough to detect perilymph in quantities as low as 2-5 microliters despite contamination by other adventitious fluids. Phase II clinical trials are also proposed: 1) assuming that a sensitive marker for PLF is developed, a multi- institutional prospective observational study is anticipated to study the prevalence of verifiable PLF and identify diagnostic prediction criteria in patients with auditory and vestibular dysfunction. 2) to address uncertainties regarding potential false negative results from intermittent PLF and microfistulae, a prospective randomized clinical trial involving patients with vestibular dysfunction is proposed to compare the efficacy of surgical treatment for presumed PLF with a control group treated in vestibular rehabilitation therapy. The aim of the research proposed is to understand the developmental mechanisms by which the cells of the retina achieve their specialized sub-types. glass mutations specifically remove the photoreceptor neurons of the Drosophila visual system; these cells begin to develop as neurons but fail to express photoreceptor specific genes, and later die. glass encodes a protein with five Zinc-finger domains; such proteins have been showm in other organisms to act as transcription factors. Thus glass may act directly to regulate photoreceptor cell-specific gene expression. We have shown that the glass protein binds in-vitro to sequences from enhancer elements of one such gene (a rhodopsin). glass is expressed in all the cells of the developing retina, but it is only active in the developing photoreceptors (by showing the affect on a reporter gene of glass DNA binding sites). Thus the regulation of glass activity is critical for photoreceptor cell development and this occurs at two levels: at transcription, and after translation (by incoming positional signals). To pursue this dual regulation we will conduct two projects: 1) To understand glass transcriptional regulation we will undertake a functional analysis of the glass gene promoter by mutational studies. 2) To identify novel genes that interact with glass we will use a series of genetic screens and the first will be for dominant enhancers and suppressors of weak glass areles. Preliminary screens have successfully tested the feasibility of this approach (we have recovered three such mutations). Once isolated we will characterize the effects of our novel mutations and in the long term, clone the genes responsible to study their molecular functions.