慢性化脓性中耳炎对内耳听觉功能的影响

为探讨慢性化脓性中耳炎影响内耳听觉功能的机理.统计了200例、240耳慢性化脓性中耳炎(COM)患者的病史、纯音测听、乳突拍片和CT扫描的结果;并对中耳病变与耳聋性质及程度的关系,进行分析.制备化脓性中耳炎豚鼠12只、15耳,在光镜及电镜下观察耳蜗Corti′s器外毛细胞的病理变化.结果:本组患者合并感音性聋(SNHL)107耳,占44.6%;病史5年以内及5年以上SNHL合并率分别为28.5%和63.6%,差异有高度显著性(P＜0.01);中耳病变Ⅰ型及Ⅱ～Ⅲ型COM患者SNHL合并率为27.9%和85.7%,差异有高度显著性(P＜0.01);观察COM豚鼠耳蜗Corti′s器外毛细胞有病理损害.COM病史越长,中耳病变越重,耳蜗外毛细胞损害越重,SNHL合并率越高.     

豚鼠中耳感染致Corti器P物质受体阳性细胞的表达

目的　应用P物质受体 (substancePreceptor,SPR)的多克隆抗体观察豚鼠中耳急性感染后内耳Corti器SPR阳性细胞的表达。方法　豚鼠一侧耳经鼓膜注射 1× 10 8个 /L金黄色葡萄球菌液 ,制备急性化脓性中耳炎模型 ,3d后灌注固定 ,取耳蜗基底膜铺片 ,行SPR免疫组织化学染色。结果　光镜下发现中耳急性感染后豚鼠耳蜗基底膜表达一种非正常耳蜗组织细胞本身的SPR阳性细胞 ,这些细胞与内、外毛细胞 ,支持细胞 ,血管内皮细胞 ,螺旋神经节细胞等在形态部位上存在明显的差异。阳性细胞呈散在分布 ,主要集中在基底膜的游离缘 ,体积较大 ,约为红细胞的 6～ 12倍 ,每个细胞均有多个突起 ,形成类神经元状。细胞呈SPR强阳性。在阳性细胞的胞质内可见空泡或颗粒状的结构。结论　中耳急性感染可诱发豚鼠Corti器表达非正常基底膜所有的SPR阳性的非特异性反应细胞 ,这种细胞可能参与启动或诱发内耳的免疫反应     

热休克蛋白70在顺铂所致豚鼠耳蜗损伤中的表达以及磷霉素钠的保护作用

目的 通过豚鼠在体实验观寨磷霉素钠对顺铂耳毒性的拮抗作用以及热休克蛋白70在耳蜗中的表达情况。方法豚鼠28只,随机分为4组:正常对照组6只;单独应用磷霉素钠组6只,磷霉素钠500mg/Kg,1日1次肌注;单独应用顺铂组8只,一次性给予顺铂15mg/Kg腹腔注射;磷霉素钠保护组8只,先肌肉注射磷霉素钠500mg/Kg,间隔30min后腹腔注射顺铂15mg/Kg。4天后处死所有动物。给药前和处死前各测听觉脑干诱发电位(ABR)1次。断头后取出听泡,固定、脱钙,常规石蜡包埋、切片。HSP70单克隆抗体免疫组化染色,光镜下观察阳性表达部位,图象处理分析系统测定灰度值。结果 单纯磷霉寨钠组ABR阈值无改变;顺铂组阈值明显升高,幅度达37.1±19.9 dBspl,P<0.001;磷霉素钠保护组阈值仅提高8.1±9.6 dBspl,同对照组相比P>0.05。HSP70表达分布在耳蜗中的Corti氏器、螺旋神经节、血管纹等处,顺铂损伤后Corti氏器和螺旋神经节细胞呈强阳性表达。从灰度值来看对照组、顺铂组与磷霉素钠保护组两两比较差异均有显著性,P<0.01或0.05。结论磷霉橐钠在一定程度上可以拮抗顺铂的耳毒性,减小ABR阈值的提高。顺铂损伤后耳蜗Corti氏器和螺旋神经节细胞HSP70呈强阳性表达,表达量与耳蜗的损伤程度一致。     

强次声波对豚鼠Corti器超微结构的损伤

目的 观察强次声波对豚鼠Corti器超微结构的损伤情况。方法 将豚鼠置于频率8Hz、强度为135dBSPL的次声声场中连续暴露90min。应用扫描电镜分别观测强次声波暴露后即刻（2h内）、2天和7天时动物Corti器超微结构的变化，计算各组耳蜗的受损率，与对照组进行比较。结果 扫描电镜下见各实验组动物Corti器感觉毛细胞纤毛缺失、散乱、倒伏，表皮板等结构均有不同程度的损伤。在存活较长时期后，还可见到纤毛融合．部分细胞的表皮板破裂，以及支持细胞分离，细胞溶解，形成空洞。耳蜗受损率分别为70％～80％。结论 强次声波可引起豚鼠Corti器超微结构不同程度的损伤。     

聚天冬氨酸抑制庆大霉素致耳蜗自由基产生的实验研究

目的观察聚天冬氨酸(polyasparticacid,PAA)对庆大霉素(gentamicin,GM)致耳蜗组织氧自由基产生的影响,进一步探讨PAA对GM耳毒性拮抗作用机制.方法将88只豚鼠随机分为Ⅰ组-单用GM、Ⅱ组-PAA+GM、Ⅲ组-单用PAA、Ⅳ组-单用生理盐水,采用电子顺磁共振法(electronparamagneticresonancespectroscopy,EPR),直接探测在体给药1、5、10d后,豚鼠耳蜗组织羟自由基(OH-)的产生量;同时,记录ABR,用透射电镜观察耳蜗Corti器毛细胞溶酶体形态改变.结果①给药1dⅠ、Ⅱ组耳蜗组织EPR积分值有升高趋势;ABR阈值4组间差异无显著性(P>0.05);耳蜗Corti器毛细胞溶酶体形态无明显改变.②给药5dⅠ组耳蜗组织EPR积分值(37.74±4.10)较其他3组明显升高(P<0.01),给药10d4组间耳蜗组织EPR积分值差异无显著性(P>0.05);超微结构Ⅰ组耳蜗Corti器毛细胞皮板下溶酶体改变给药10d较给药5d更明显,可见溶酶体聚集数量增多,体积明显增大;Ⅰ组ABR阈值随用药时间延长而升高.结论PAA对GM致耳蜗组织氧自由基的产生有抑制作用,从而拮抗了GM所致溶酶体磷脂沉积病态及耳蜗毒性的发生.     

应用微波技术行后半规管阻塞术的实验研究

目的 :探讨应用微波技术行后半规管阻塞术的可行性。方法 :应用微波技术对 2 0只豚鼠行后半规管阻塞术 ,观察术前、术后豚鼠眼震电图 (ENG) ,听性脑干反应 (ABR)及形态学的变化。结果 :术后第 1天豚鼠正弦摆动刺激时术侧眼震反应减弱 ;术后第 3天双侧眼震恢复对称 ;术后 30 d冰水实验结果显示两耳能引出对称性眼震 ;术后 ABR阈值无明显改变 ;术后 6 0 d病理切片证实后半规管完全阻塞而耳蜗 Corti器和壶腹嵴正常。结论 :应用微波技术行后半规管阻塞术是对后半规管阻塞术的一种有意义的改进方法。     

豚鼠耳蜗Corti器外隧道的结构

目的 观察豚鼠耳蜗Corti器外隧道的解剖特征.方法 健康杂色豚鼠11只,共22耳,借助火棉胶切片(5只10耳)和扫描电镜(6只12耳)观察和分析豚鼠耳蜗Corti器外隧道的解剖.结果 豚鼠耳蜗Corti器外隧道的内侧壁在耳蜗基底回和第一回由第三排外毛细胞和Deiter细胞的指状突起构成,外侧壁和顶壁由Hensen细胞构成.自耳蜗基底回逐渐向顶回,Deiter细胞的指状突起逐渐移向外侧,形成外隧道的外侧壁和顶壁.外隧道的内侧壁由第三排外毛细胞构成,Deiter细胞指突移向Hensen细胞构成外隧道的外侧壁和顶壁,外隧道腔隙逐渐缩小.第三、四回的外隧道完全被Deiter细胞所充填,Deiter细胞指突和Hensen细胞密切接触.结论 豚鼠耳蜗Corti器外隧道的这种结构特征在研究Corti器的微细结构方面增加了新的认识.     

小动物内耳影像学研究进展

<正>内耳又称为迷路,内部结构复杂,由骨迷路和膜迷路构成。内耳解剖结构的完整是听觉形成的物理基础。耳蜗是内耳的重要结构之一,耳蜗对声音具有高度敏感性和高度选择性,任何引起耳蜗结构和功能改变的疾病都会导致不同程度的听力下降。耳蜗中最重要的是柯蒂氏器(organ of Corti),Corti氏器由听毛细胞支持细胞和和盖膜所组成[1]。耳蜗内有听觉通路第一级神经元,一级神经元的树突始于Corti氏器听毛细胞的基底,轴突延伸成为耳蜗神经,     

新霉素损伤后小鼠耳蜗毛细胞的改变[论著]

目的　观察新霉素对小鼠听功能及耳蜗毛细胞形态学变化的影响。方法　选取C57BL/6小鼠16只在出生后第8天开始连续10 d皮下注射硫酸新霉素，对照组（10只）皮下注射等量生理盐水。停药后1、4、8、12 w进行听性脑干反应（ABR）检测。每次ABR检测后，新霉素组随机取3只小鼠，对照组随机取2只小鼠，处死后，取耳蜗进行冰冻切片，用免疫荧光方法观察耳蜗Corti器及毛细形态学变化。结果　新霉素可造成小鼠耳蜗毛细胞严重损伤，且小鼠ARB阈值显著增高，不可恢复；新霉素对耳蜗Corti器毛细胞的损伤顶圈最轻，中圈次之，底圈最重，且随着时间推移Corti器形态结构破坏逐渐加重且不可自我修复。结论　硫酸新霉素造成小鼠Corit器毛细胞的损伤是不可逆的。     

早期气导听觉剥夺对幼鼠听觉发育的影响

目的研究早期气导听觉剥夺对大鼠听性脑干反应(ABR)和Corti器基底膜发育的影响。方法 SD仔鼠共60只,随机分为听觉剥夺组及对照组,每组30只。听觉剥夺组行双外耳道填塞后饲养于密闭隔声室,对照组在正常声音环境饲养,两组动物分别于出生后21、28、35、42及56天时行ABR检测,42及56天时应用扫描电镜观察Corti器基底膜发育情况。结果对照组大鼠ABR反应阈随年龄增长呈下降趋势,56天时为24.17 dB SPL,基本达到正常成年大鼠水平,听觉剥夺组大鼠随饲养时间延长ABR反应阈较对照组明显增高,35天时增高至39.47 dB SPL,去除双外耳道填塞物后ABR反应阈无明显恢复。扫描电镜可见对照组Corti器基底膜外毛细胞纤毛排列整齐,听觉剥夺组基底膜外毛细胞表面形成大量细小囊泡,纤毛末端膨大、融合。结论出生后早期气导听觉剥夺可造成大鼠Corti器基底膜发育异常以及听觉障碍,表明气导听觉传入在听觉发育过程中起着十分重要的作用。     

经鼓室注射内耳钆造影显影不良原因初探

目的 总结分析经鼓室注射内耳钆造影显影不良的可能原因,探讨可行的解决方案。 方法 分析经鼓室注射内耳钆造影的356例患者(共706耳)病历资料,发现存在显影不良55例(共70耳),分析显影不良可能存在的原因。 结果 55例(共70耳)中表现为显影浅淡16耳,表现为充盈欠佳14耳,不显影/未见造影剂充盈40耳。影像学提示单纯合并同侧中耳炎者39耳,考虑慢性中耳炎或隐匿性中耳炎,其中出现显影不良11耳(约28.2%,11/39);单纯合并鼻窦炎者32耳,出现显影不良4耳(约12.5%,4/32);同时合并中耳炎和鼻窦炎者3耳,出现显影不良者1耳(约33.3%,1/3);影像学未提示合并中耳炎/鼻窦炎者632耳,其中出现显影不良54耳(约8.5%,54/632)。合并中耳炎与未合并相比差异有统计学意义,合并鼻窦炎、同时合并中耳炎鼻窦炎与未合并比较差异无统计学意义。 结论 经鼓室注射内耳钆造影时,排除近期中耳炎病史后,慢性中耳炎或隐匿性中耳炎仍可能是显影不良的影响因素。通过对造影各个环节中其他可能存在的影响因素进行进一步探讨,建议根据可能存在的原因尽量规避,以提高内耳钆造影的成功率。     

Predisposing factors for inner ear hearing loss association with chronic otitis media

The aim of the present study was to investigate the consequences of chronic otitis media on inner ear function. Retrospective analysis of conventional pure-tone audiometry tests was carried out on 344 patients who were scheduled for surgical treatment of unilateral chronic otitis media without other risk factors for sensorineural hearing loss. Bone conduction thresholds of diseased ears were compared with those of contralateral, non-diseased ears. Selected clinical features were assessed among diseased ears to examine possible influences on inner ear function. Mean bone conduction threshold differences varied from 0.6 dB at 0.5 kHz to 3.7 dB at 4 kHz. These differences augmented with increasing duration of middle ear disease. Impaired hearing by bone conduction thresholds of diseased ears correlated with increased age at every frequency and with an interruption of the ossicular chain only at higher frequencies. The severity of sensorineural hearing loss correlated with longer duration of middle ear disease. Thus, surgical treatment of dry and apparently stable tympanic membrane perforation is warranted.     

Predisposing factors for inner ear hearing loss association with chronic otitis media

The aim of the present study was to investigate the consequences of chronic otitis media on inner ear function. Retrospective analysis of conventional pure-tone audiometry tests was carried out on 344 patients who were scheduled for surgical treatment of unilateral chronic otitis media without other risk factors for sensorineural hearing loss. Bone conduction thresholds of diseased ears were compared with those of contralateral, non-diseased ears. Selected clinical features were assessed among diseased ears to examine possible influences on inner ear function. Mean bone conduction threshold differences varied from 0.6?dB at 0.5?kHz to 3.7?dB at 4?kHz. These differences augmented with increasing duration of middle ear disease. Impaired hearing by bone conduction thresholds of diseased ears correlated with increased age at every frequency and with an interruption of the ossicular chain only at higher frequencies. The severity of sensorineural hearing loss correlated with longer duration of middle ear disease. Thus, surgical treatment of dry and apparently stable tympanic membrane perforation is warranted.     

Labeling of the glucocorticoid receptor and Na,K-ATPase in a rat otitis media model

HYPOTHESIS: Glucocorticoid hormones exert an influence on the inflammatory response of the middle ear during acute otitis media. Rats with experimentally induced purulent otitis media were given either glucocorticoid hormones in excess or a glucocorticoid hormone blocker that deprived the animals of the hormone. BACKGROUND: Acute otitis media is a common inflammatory disease among children. Streptococcus pneumoniae is the most usual causative agent. The standard treatment today is phenoxymethylpenicillin. The role of glucocorticoid hormones in inflammatory reactions in the middle ear has been widely debated. METHODS: In an otitis media model, a suspension of pneumococci was inoculated into the bulla of the rat, after the animals were pretreated with either a dose of corticosteroid hormones or the glucocorticoid receptor blocking agent RU 486. Rats with induction of otitis media only, but no pretreatment, were used as control subjects, as were the left control-operated ears of all rats. The inflammatory response in the inner ear and in the middle ear was evaluated. The presence of glucocorticoid receptors and the enzyme Na,K-ATPase was investigated with immunohistochemistry. RESULTS: The inflammatory response in the animals with untreated otitis media and in the group with otitis media in rats pretreated with the receptor blocker was much more extensive than in the group of animals pretreated with corticosteroids. In the corticosteroid-treated group, the tympanic membrane and the mucous membrane of the middle ear were less edematous, but the middle ear cavity contained more pus. Only a few lymphocytes were found in the inner ears of these rats. When the inner ear was labeled with antibodies against glucocorticoid receptors, there seemed to be no difference between the labeling patterns in the three groups. This was also the case for antibody labeling against Na,K-ATPase. CONCLUSION: The present results indicate that the reaction in the middle ear mucous membrane is more pronounced in rats that had been pretreated with the hormone receptor blocking drug. An increase of corticosteroid hormone levels during the inflammatory process seem to diminish the reaction in the tympanic membrane and the middle ear mucosa. Neither the hormone receptor blocking drug nor the steroid hormones change the content of glucocorticoid receptors and Na,K-ATPase in the inner ear in the otitis media rat model.     

Image analysis of the inner ear with CT and MR imaging: pre-operative assessment for cochlear implant surgery]

Recent progress in magnetic resonance imaging (MRI) has made it possible to obtain detailed images of the inner ear by delineating the lymphatic fluid within the labyrinth. We analyzed CT scans and MR images in 70 ears manifesting profound deafness owing to inner ear lesions and compared their detective ability for inner ear lesions. The following results were obtained. 1) CT scan examination showed slight to extensive ossification of the labyrinth in six ears (9%), whereas MRI examination revealed low to absent signal intensity of the inner ear in nine ears (13%). Therefore, it was concluded that MRI is more sensitive in detecting abnormalities of the inner ear than CT scan. 2) MRI provided useful information as to whether the cochlear turn is filled with lymphatic fluid or obstructed. This point was one of the greatest advantages of MRI over CT scan. 3) Abnormal findings in either or both the CT scan and the MRI were detected in suppurative labyrinthitis occurring secondary to chronic otitis media, bacterial meningitis and in inner ear trauma. However, such abnormal findings were not detected in patients with idiopathic progressive sensorineural hearing loss, ototoxicity or sudden deafness. These findings should be taken into consideration in pre-operative assessment of cochlear implant candidates.     

中耳细菌感染诱导的急性期HSP-70相关反应的免疫组织化学研究

探讨中耳细菌感染急性期时 ,哺乳动物中、内耳热休克反应的部位 ,以及中耳细菌感染诱生的热休克蛋白是否可能引发内耳自身免疫损伤。运用中耳注射肺炎克雷伯杆菌制成豚鼠中耳急性感染动物模型 ,分别于接种后第 1、3、5、7天处死动物取材。应用免疫组化技术 ,研究了中耳粘膜和耳蜗表达热休克蛋白 70 (HSP- 70 )的部位。结果表明 :正常状态下 ,中耳粘膜表层的上皮细胞和内耳膜迷路血管纹、螺旋韧带、Corti氏器均有弱的阳性反应 ,感染应激后 ,上述同样部位均有强的阳性显色。不同取材时段显示的阳性位置无差异。说明在中耳急性细菌感染期 ,中耳粘膜和内耳组织均表达了同源 HSP- 70蛋白分子 ,这些同源 HSP- 70为引发内耳自身免疫损伤提供了物质基础。     

Effects of Burow's solution as an ear drop on Intractable chronic suppurative diseases of the external ear canal and middle ear

OBJECTIVE: Burow's solution was developed by Karl August von Burow in the 19th century as an ear drop. Thorp et al. reported its excellent effect on chronic suppurative otitis media in 1998. We applied Burow's solution to intractable chronic purulent diseases of the external and middle ear in the 12 months from February 2001. SUBJECTS: Subjects were 25 ears of 21 patients--35-79 years old, 10 men's ears and 15 women's ears--whose disease has continued for an average of 3.78 years. Diseases and patients are as follows: 1) 11 ears with postoperative mastoid cavity problems, 2) 7 ears with chronic external otitis and chronic eczema of the external ear canal, 3) 7 ears with fungal otitis externa, 4) 6 ears with chronic otitis media with perforation, and 5) 2 ears of chronic granulated myringitis. METHOD: The solution was dropped into the ear canal once a day for 10 min or cotton balls soaked with the solution were applied to the canal wall. Criteria of efficacy were divided into cured, effective and unchanged. RESULTS: 8/11 ears with a cavity problem were cured (72.7%) and 3 ears showed treatment to be effective. All 7 ears with chronic external otitis and chronic eczema were cured. All 7 ears with fungal otitis externa were cured. 4/6 ears (66.6%) with chronic perforated otitis media were cured and 1 ear each showed treatment to be effective and 1 unchanged. Two chronic granurated myringitis were cured. Twenty of 25 ears (80%) were cured. Pre- and posttreatment audiometries showed almost the same. The cure appeared within 3 days to 3 weeks. The solution was not effective against mucoid secretion, cholesteatoma, or residual mastoid cells. The day after treatment, serous secretion appears temporarily. CONCLUSION: Burows's solution is seemed to be very effective and nonototoxic as an otic drop for treating suppurative ear diseases.     

Permeability of the normal round window membrane to Haemophilus influenzae type b endotoxin.

Sensorineural hearing loss associated with otitis media may be due to passage of ototoxic substances such as bacterial toxins and antibiotics, from the middle ear into the inner ear. The round window membrane is the most likely route for such transport. The aim of this study was to analyze the extent of endotoxin passage through the normal round window membrane. The round window membranes of 19 chinchillas were exposed in vivo to Gelfoam soaked in purified Haemophilus influenzae type b endotoxin at a concentration of 45,000 endotoxin units per ml (EU/ml) during 3 to 24 h. Endotoxin levels in the perilymph were measured with Limulus Amaebocyte Lysate or Quantitative Chromogenic Limulus Amaebocyte Lysate. Endotoxin was detected in half of the inner ears at concentrations close to the detection limit (approximately 4 EU/ml). The results suggest that the normal round window membrane efficiently protects the inner ear against the passage of bacterial endotoxins from the middle ear cavity. It is unlikely that endotoxin at concentrations found in the middle ear secretion during otitis media can traverse the round window membrane in sufficient amount to cause inner ear deterioration.     

Cochlear polyamines: Markers of otitis media-induced cochlear damage

High-performance liquid chromatography (HPLC) set to the femtoliter sensitivity level was used to identify and quantify the polyamines spermidine and spermine as well as the diamine putrescine in the different tissues of the inner ears of guinea pigs with experimentally induced otitis media. The tissues examined were the lateral wall (stria vascularis and the spiral ligament), the organ of Corti, and the cochlear nerve. The difference in polyamine profile in the different tissues of the control noninfected guinea pigs suggests a relation to the particular function of each of these tissues. The difference in polyamine profile in infected different inner ear tissues compared to controls encourages the assumption that the polyamines may be involved in a repair process of the inner ear after injury and that they may be considered as biochemical markers for inner ear damage secondary to acute otitis media.     

Gram-negative pathogen Klebsiella oxytoca is associated with spontaneous chronic otitis media in Toll-like receptor 4-deficient C3H/HeJ mice

CONCLUSION: This report confirms the presence of gram-negative Klebsiella bacteria in the middle ear of the C3H/HeJ mouse by culture, polymerase chain reaction (PCR), and electron microscopy. Identification of the bacterial pathogen supports the C3H/HeJ mouse as an excellent model for spontaneous chronic otitis media and its effects on the middle and inner ear. OBJECTIVES: The C3H/HeJ mouse has a single amino acid substitution in its Toll-like receptor 4, making it insensitive to endotoxin and suppressing initiation of the innate immune system. This study explored the bacteriology of the resultant middle ear infection by culture, PCR, histology, and electron microscopy. MATERIALS AND METHODS: Twelve-month-old C3H/ HeJ mice were screened positive for spontaneous otitis media. Tympanocentesis and blood cultures of mice were carried out under sedation. Middle ear aspirate material and blood samples were then sent for culture and PCR. Mice were then sacrificed for bright-field and electron microscopy analysis. RESULTS: All tympanocentesis and blood specimens grew gram-negative Klebsiella oxytoca, which was confirmed by PCR. Histopathology confirmed an intense inflammatory reaction and gram-negative bacteria in the middle and inner ears. Electron microscopy of the middle ears revealed abundant rod-shaped Klebsiella bacteria, both free and being engulfed by neutrophils.