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自身免疫性内耳病的实验研究
许多感音神经性聋与免疫因素有关,部分突聋、梅尼埃病、耳硬化症、慢性中耳炎、颞骨创伤,以及一些伴发听力丧失的全身免疫性疾病都存在内耳免疫损伤因素.为了探讨Ⅱ型胶原(type Ⅱ collagen,C Ⅱ)在自身免疫性内耳病(autoimmune inner ear disease,AIED)AIED发病中所起的作用,本实验用鸡Ⅱ型胶原建立豚鼠自身免疫性内耳病模型,用免疫组织化学技术观察模型建立初期细胞间粘附因子-1(intercellular adhesion molecule 1,ICAM-1)的表达情况,并从功能、形态方面评价动物模型的内耳损伤情况.
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基因芯片在内耳基因功能与突变检测中的应用
基因芯片又称DNA芯片,寡核苷酸微阵列等,是在人类基因组计划实施过程中出现并发展起来的.它融合了多领域的各项技术,具有高通量、高集成、微型化和自动化的特点,能够高效平行地处理和应用日益庞大的基因组信息,被称为继单克隆抗体技术和PCR技术之后生命科学中的又一重大技术创新[1],在序列分析、新基因的发现、基因表达谱分析、基因诊断及构建单核苷酸多态性(single nucleotide polymorphism,SNP)图谱等方面发挥着日益重要的作用.
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纳米基因载体及其在内耳基因治疗中的研究进展
基因治疗是借助载体把目的基因高效且安全地输送到特定的病变细胞或组织中,从而进行疾病治疗的一种方法.内耳疾病在耳鼻喉科的发病率较高,严重影响人们的健康和生活质量,且目前尚无确切的治疗方法.随着分子生物学的发展,内耳基因治疗研究为攻克这一顽症带来希望.但目前阻碍内耳基因治疗的一大难题就是缺乏理想的基因载体[1].
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内耳疾病的局部药物治疗
内耳疾病(inner ear disorder)特指发生于迷路范围内的听觉(耳蜗)与平衡(前庭)器官病变,是一类严重影响人类生活质量的疾病.鉴于内耳组织结构的微观特性与精细程度,内耳疾病的临床病理与症状十分复杂,某些病变的发病原因及机制迄今不明.对于内耳疾病的治疗方法繁多,包括药物治疗、手术治疗与康复治疗.随着给药技术及药物剂型的发展,局部给药已经成为治疗内耳疾病的重要方法,日益引起人们的关注.与传统的全身给药方式相比局部给药所具有的优势包括:①直接治疗患耳,对全身各系统无明显影响;②内耳局部可达到较高的药物浓度,使治疗更为有效;③避免全身给药带来的毒副作用;④可以达到缓释[1-2]、控释[3]及靶向[4]给药,因而具有明显的临床优势.
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亲和层析法纯化内耳髓鞘蛋白零
在内耳的螺旋神经节及蜗神经的髓鞘上存在髓鞘蛋白零(myelin protein zero,P0蛋白),相对分子量为30 000[1].内耳P0蛋白可能是某些自身免疫性内耳病(autoimmune inner ear disease,AIED)的自身抗原[1,2].现介绍用亲和层析法纯化内耳P0蛋白,为进一步探讨P0蛋白在自身免疫性内耳病中的作用提供实验基础.
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制备性聚丙烯酰胺凝胶电泳分离与纯化豚鼠内耳抗原
粗制内耳抗原(crude inner ear antigen,CIEAg)广泛用于自身免疫性内耳病(autoimmune inner ear disease,AIED)的研究,包括建立动物模型和作为抗原检测AIED患者血清自身抗体[1,2]。但CIEAg成分复杂,其中何种亚组份起关键作用尚不清楚。我们通过制备性聚丙烯酰胺凝胶电泳(preparative polyacry-lamide gel electrophoresis,简称制备性PAGE)分离纯化豚鼠CIEAg的主要亚组份,为深入研究AIED的发病机制和寻找内耳特异性抗原奠定基础。 一、材料和方法 1.豚鼠CIEAg的制备:取200~300 g耳廓反射正常的豚鼠(华中科技大学同济医学院实验动物部提供),参照肖红俊等[3]报道的方法提取。考马斯亮蓝G-250法测定其浓度。 2. 凝胶电泳方法:①分析性聚丙烯酰胺凝胶电泳(analytical polyacrylamide gel electrophoresis,以下简称分析性PAGE)凝胶厚1.25 mm,浓缩胶和分离胶的浓度分别为4%和10%;加样量为4~32 μg,蛋白质分子量标准加样20或40 μg;样品进入分离胶前恒电流10 mA,进入分离胶后恒电流20 mA,15℃恒温电泳;常规考马斯亮蓝R-250染色和脱色,并对脱色后凝胶进行薄层扫描;②制备性PAGE 凝胶厚3 mm,加样量为2~3 mg;样品进入分离胶前恒电流40 mA,进入分离胶后恒电流50 mA,15℃恒温电泳;电泳结束后,快速考马斯亮蓝R-250染色液加温染色40~50 min,然后在含有5%甲醇、10%冰乙酸的脱色液中加温脱色10 min左右[4],其余同①;③制备性PAGE和分离亚组份的回收电泳同②;电泳结束后自板胶的中央纵向切下宽1 cm左右的窄带,进行快速染色和脱色。将染色后的凝胶条用蒸馏水冲洗后复位,根据染色凝胶蛋白区带的位置,从未染色的凝胶上切下主要蛋白区带冰浴匀浆。-50℃保存备用。
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膜迷路积水造模中各种内淋巴囊手术入路评介
膜迷路积水(endolymphatic hydrops,EH),又称内淋巴积水,1938年由Hallpike和Cairns发现其为梅尼埃病(Meniere’s disease,MD)的基本病理变化。随着对膜迷路积水的不断研究探讨,目前认为膜迷路积水是一些由于内耳损伤所导致的不同疾病的共同病理改变,包括梅尼埃病、伴眩晕的突发性耳聋(sud-den onset sensorineural hearing loss)、迟发性膜迷路积水(delayed endolymphatic hydrops,DEH)、急性低频感音神经性听力损失(acute low tone sensorineural hear-ing loss)、Lermoyez综合征(Lermoyez’syndrome)及自身免疫性内耳病(autoimmune inner ear disease)等疾病。其中MD、DEH及伴眩晕的突发性耳聋已经通过影像学直观地观察到膜迷路积水表现[1-3]。由于与膜迷路积水相关疾病的发病机制仍不明化,临床表现复杂化,实验检查多样化,且膜迷路积水在相关疾病中因果关系仍不确定。为更好探讨膜迷路积水的发生、发展及演变过程,国内外学者纷纷建立了各种膜迷路积水动物模型。
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Diabetes mellitus (DM) is a chronic systemic disease characterized by hyperglycemia, with various patho-genic mechanisms. From absolute or relative insulin deficiency, patients with DM often demonstrate vari-ous levels of metabolic disorders. Major clinical manifestations of DM include metabolic disorders, vascu-lar lesions, circulatory disturbances and neurologic complications. Along with advances in DM research, re-ports of DM related tinnitus and hearing impairment have increased continuously. Research on DM related auditory system dysfunction has focused on cochlear microcirculation, cellular homeostasis, genetics and ag-ing. Cochlear microcirculation plays an important role in cochlear physiology and its disorders are associat-ed with many inner ear diseases. Ischemia and subsequent reperfusion seen in cochlear microcirculation dis-orders are important factors in hearing damage. Understanding cochlear microcirculation and structural as well as functional changes in DM patients with hearing loss and their causal factors will help reveal patho-genic mechanisms in diabetic hearing loss and provide new ideas in developing interventions and preventing damages caused by diabetes.
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自身免疫性内耳疾病
1 前言多种原因可以引起感音神经性耳聋.突然发生的渐进性感音神经性耳聋对患者来说非常苦恼.自身免疫性内耳疾病(autoimmune inner ear disease)为一很罕见的诊断,因为在评估突然发生的感音神经性耳聋,患者时常将自身性内耳疾病这一诊断自各种不同诊断中遗漏掉.由于自身免疫性内耳疾病所致的听力下降及前庭症状是有可能治愈的,因此不能正确诊断这一疾病令人遗憾.
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高压氧对突发性耳聋的康复效果观察
Background: The cause of paroxysmal deafness is unknown.It is a nerve deafness that has mostly hearing handicap and harried onset.At present,the disease is usually deemed incurable if it continues for three months.Many scholars consider thathandicap of blood circulation in inner ear is an important factor leading to paroxysmal deafness.Hyperbaric oxygenation improves oxygenic content and press in blood.Objective: The effects of hyperbaric oxygenation on rehabilitation of paroxysmal deafness were investigated.Design: 105 patients with paroxysmal deafness were randomly divided into treatment group(n=41)and control group(n=43).Treatment group included 30 patients single ear with deafness of unilateral ear.Control group included 28 patients with deafness of unilateral ear.Unit: Liaocheng People's Hospital.Subject: We collected 105 patients with paroxysmal deafness from May 1999 to Dec 2000.84 patients were male,21 patients were female,aged 7~ 60 years(average 31.9 years).
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大鼠内耳胚胎和出生后的组织学研究(英)
The mouse labyrinth is an excellent model for study-ing the morphogenesis and cytodifferentiation of the mam-malian inner ear' s postnatal developmental periods andthe availability of numerous genetic mutants with a variety of inner ear abnormalities. There are many genes affectingthe inner ear of the mouse, and some of these lead to le-sions which resemble lesions occurring in man.
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大鼠内耳发育过程表皮生长因子的表达(英)
Since the two independent discoveries in 1987 indi-cated that hair cells within cochlea of bircls were capableof spontaneous regeneration, recent observations havedemonstrated that, in response to injury, mammals pos-sess a limited capacity of regenerating inner ear sensoryepithelia, but the mechanism of regeneration is still tm-known. Lewfebvre (1993) and Lambert (1994) reportedrespectively that retininoic acid (RA) and transforminggrowth factor alpha (TGF-α) could stimulate hair cell re-generation in the mammalian inner ear. These studies in-dicated that the cellular factors participating the normaldevelopment of cells could play a critical role in the re-generative process of hair cells.Epidermal growth factor (EGF) plays a critical rolein proliferation and differentiation of normal cells and tis-sues and it is extensively expressed in many mature andembryonic tissues.
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突发性感音神经性聋的免疫学研究进展
突发性感音神经性聋(sudden sensorineural hearing loss,SSNHL)表现为在数小时至3日、至少连续3个频段听力下降超过30 dB.该病仅有10%~15%的患者能发现病因,预后与起病时听力损伤程度、是否及时就诊、是否伴有眩晕等因素相关.目前该病无肯定发病机理,有病毒感染、内耳供血障碍及内淋巴膜破裂三种假说.1979年McCabe[1]依据听力下降患者经过免疫抑制治疗后听力有明显改善,命名了自身免疫性感音神经性聋(autoimmune sensorineural hearing loss,ASNHL),后又提出自身免疫性内耳病(autoimmune inner ear disease,AIED),此后对免疫与内耳疾病的研究日益深入广泛.Wilson等[2]将中度听力下降SSNHL患者给予皮质激素与安慰剂,结果给予皮质激素组听力恢复率为78%,明显高于安慰剂组的38%,提示免疫反应在SSNHL发病中占重要地位.而许多自身免疫性疾病如Cogan综合征、颞动脉炎、Wegener肉芽肿、结节性多动脉炎(PAN)、系统性红斑狼疮(SLE)等,在病程中可有SSNHL表现,进一步说明免疫与SSNHL密切相关.本文将SSNHL的免疫学研究进展作一综述.
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Ⅱ型胶原与自身免疫性内耳病
1979年,McCabe发现某些感音神经性聋的临床表现及实验室检查特征提示其为自身免疫性疾病.其特征为:数周或数月内发展的渐进性聋(而非数小时、数天或数年);感音神经性聋通常为双侧或非对称性;耳蜗常受累;常伴有面瘫和前庭功能不良;可有鼓膜、中耳和乳突组织破坏;对大剂量类固醇激素和环磷酰胺疗效好.McCabe将此类疾病定义为:自身免疫性内耳病(autoimmune inner ear disease,AIED)[1].现认为AIED可理解为两重范畴:一为狭义,如McCabe所描述,即AIED为一独立的疾病实体;一为广义,即与自身免疫相关的一组内耳疾病,包括梅尼埃病、耳硬化症和狭义范畴的AIED等.本文所指为其广义范畴.
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自身免疫性内耳病的临床诊断与治疗
1979年McCabe首先提出自身免疫性感音神经性聋(au-toimmune sensorineural hearing loss,AISNHL).以后的研究发现自身免疫性损害不仅累及耳蜗,也可累及前庭,故称之为自身免疫性内耳病(autoimmune inner ear disease,AIED).