一氧化氮合成酶和心钠素在豚鼠耳蜗的分布

目的:观察豚鼠耳蜗局部心钠素(ANP)和一氧化氮合成酶(NOS)免疫组化反应产物的分布及其相互关系,为研究ANP和NOS在豚鼠耳蜗局部血流、淋巴以及神经调节中的相互作用提供形态学研究的依据。方法:采用免疫荧光组织化学双标法观察ANP和NOS在正常豚鼠耳蜗的分布特征。结果:在耳蜗各转螺旋动脉和血管纹均发现ANP和NOS双阳性表达,螺旋缘、螺旋韧带和Corti器,耳蜗神经节细胞有ANP和NOS双阳性染色。结论:ANP和NOS的分布特点显示,二者在内耳血流调节,内、外淋巴平衡调节以及神经信号传递等方面可能存在密切的相互作用。     

胎儿耳蜗外侧壁微血管构筑的解剖学研究

目的 观察人类胎儿耳蜗外侧壁微血管形态、分布及生理学意义。方法 应用墨汁灌注和微血管铸型技术观察30例胎儿耳蜗外侧壁微血管形态。结果 耳蜗外侧壁由外放射状动脉供血,外放射状动脉走行于耳蜗前庭阶顶部,在耳蜗外侧壁形成四组毛细血管网,遍布耳蜗各回,分别为:血管纹血管网、前庭阶血管网、螺旋凸血管网和螺旋韧带深部血管网。耳蜗侧壁的静脉血流入鼓阶静脉。结论 26周龄以上胎儿耳蜗外侧壁微血管已发育完善,分布与其生理功能密切相关。     

核因子κB p65和p50在正常豚鼠耳蜗中的定位表达

目的:研究核因子κB(nuclear factor-kappa B,NF-κB)的亚单位p65和p50在正常豚鼠耳蜗中的定位与表达。方法:应用免疫组织化学SP法。结果:NF-κB p65免疫反应活性细胞位于螺旋韧带Ⅰ、Ⅲ、Ⅴ型纤维细胞和根细胞,而螺旋神经节、血管纹和螺旋器均未见染色;NF-κB p50免疫反应活性细胞位于耳蜗螺旋器的内、外毛细胞和支持细胞,此外,螺旋神经节、螺旋韧带和血管纹中也有p50表达,在耳蜗的上述部位中,NF-κB p50免疫染色多位于胞质内,胞核极少。结论:NF-κB p65和p50在正常豚鼠耳蜗的多种细胞中都有表达。     

干细胞移植豚鼠耳蜗形态结构及听力的变化

背景：既往研究证明耳蜗微造孔注射病毒或非病毒载体行基因治疗对耳蜗结构及听功能的影响轻微。 目的：观察骨髓间充质干细胞通过耳蜗微造孔术移植到正常豚鼠内耳后对耳蜗结构及听功能的影响。 方法：正常豚鼠分3组,空白对照组未予任何干预;单纯耳蜗微造孔术组单纯行耳蜗微造孔术,不注射任何成分;骨髓间充质干细胞组鼓阶微造孔后注射经DAPI荧光标记的骨髓间充质干细胞。 结果与结论：单纯耳蜗微造孔术组、骨髓间充质干细胞组经耳蜗微造孔术后7 d的听性脑干诱发电位阈值均较空白对照组提高,术后28 d恢复至正常水平;耳蜗石蜡切片苏木精-伊红染色显示实验动物耳蜗内结构无明显异常改变。骨髓间充质干细胞组耳蜗切片可见荧光信号多分布于耳蜗底周的外淋巴腔(鼓阶和前庭阶),无堆积堵塞管腔情况。结果表明经鼓阶开窗行骨髓间充质干细胞移植对耳蜗形态结构和听力的影响是轻微的,干细胞移植后能在耳蜗内迁移并存活。     

豚鼠脂肪间充质干细胞植入耳聋模型修复听力的作用

背景：脂肪间充质干细胞是否是治疗因毛细胞退化、缺失所造成的感音神经性聋的福音呢？ 目的：探讨豚鼠脂肪间充质干细胞经耳蜗鼓阶途径植入感音神经性耳聋动物模型后对听力的修复作用。 方法：庆大霉素腹腔注射建立豚鼠感音神经性耳聋动物模型,耳蜗鼓阶途径植入豚鼠脂肪间充质干细胞,分别于植入后1,3周检测听性脑干反应,观察植入脂肪间充质干细胞后耳聋动物听力的变化;并追踪EDU标记的豚鼠脂肪间充质干细胞在耳蜗内的迁移及分布情况。 结果与结论：在植入后1周及3周进行听性脑干反应检测,听力较移植前逐渐好转。植入细胞后1周,细胞大多分布在外淋巴液中,部分迁移至耳蜗柯蒂器贴附于基底膜上,植入细胞后3周,细胞不仅迁移并贴附在Corti器基底膜发挥作用,而且部分迁移到蜗神经,植入时间越长,存活细胞越少。结果表明豚鼠脂肪间充质干细胞通过耳蜗鼓阶途径微孔植入,可以定向迁移并存活最终达到提高听力的目的。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

爆震后活体豚鼠耳蜗微循环的实验观察

耳蜗微循环在听觉生理中起着十分重要的作用,本文在建立活体豚鼠耳蜗微循环实验方法的基础上,观察了爆震后一小时内的耳蜗外侧壁微循环的改变,并将微循环的变化与内耳毛细胞、听力下降进行了比较,结果显示:1)正常情况下血管纹毛细血管和螺旋韧带血管血流速度稳定,无自发性管径收缩与扩张。血管纹毛细血管血     

光化学诱导豚鼠耳蜗微循环障碍的实验研究

目的建立耳蜗微循环障碍模型并探讨其损伤机制。方法用四碘四氯荧光素二钠经豚鼠股静脉注射,波长(540±30)nm的绿色光照射耳蜗,通过耳蜗切片、螺旋韧带石蜡定向包埋切片、耳蜗铺片、透射电镜等技术在不同时问段对豚鼠耳蜗病变进行观察。结果光化学诱导耳蜗微血管血栓形成,组织缺血、坏死,且随时间的延长病变逐渐加重。结论光化学诱导可建立耳蜗微循环障碍的模型,对研究内耳疾病有一定意义。     

强噪声对耳蜗组织化学影响的研究

本实验将豚鼠分为四组:一组为对照组,另三组为实验组。所有豚鼠实验前都作耳廓反射阈测试,分别暴露于150 dB、156 dB和162 dB声压级的强噪声场5分钟。噪声暴露毕,再作耳廓反射阈测试。然后斩头处死豚鼠,速取双侧颞骨,打开骨泡,暴露耳蜗,一耳作琥珀酸脱氢酶试验、一耳作糖原和核酸(RNA和DNA)试验。结果如下: 150 dB组耳廓反射阈增高3.8 dB;156 dB组近半数豚鼠(12只耳)耳廓反射阈完全消失,其余(18只耳)耳廓反射阈增高35.6 dB;162 dB组几乎全部豚鼠(28只耳)耳廓反射阈完全消失。三组实验豚鼠琥珀酸脱氢酶和糖原都有类似变化。150dB组豚鼠耳蜗螺旋器毛细胞中,这两种物质较对照组轻度损失( ～ ),而且从底部向顶部越加明显。156dB和162 dB组,各耳蜗圈螺旋器毛细胞琥珀酸脱氢酶和糖原,一致性明显损失( ～±)。三组豚鼠耳蜗螺旋器和螺旋神经节细胞核酸(RNA和DNA)都未见明显变化。本实验表明,随噪声声压级增高,听力损伤程度也越加严重。琥珀酸脱氢酶和糖原都是与能量代谢有密切关系的物质,因此可认为,噪声刺激的早期主要引起耳蜗能量代谢障碍。     

树、大鼠、豚鼠视交叉上核加压素免疫阳性神经元分布的差异性比较

目的 :观察加压素免疫阳性 (VP IR)神经元在树下丘脑视交叉上核中的分布并与其在大鼠豚鼠视交叉上核中的分布进行比较 ,探讨VP IR神经元分布的种属差异性。方法 :树、大鼠、豚鼠内固定后恒冷箱冰冻连续切片 ,免疫组织化学PAP法染色。结果 :三种动物的视交叉上核形状不同 ,VP IR神经元在三种动物视交叉上核中的分布位置、数量和密度不同 ,此外 ,在豚鼠两侧视交叉上核间以及第三脑室底部与视交叉正中背侧之间 ,还观察到一些散在分布的VP IR神经元 ,而树和大鼠相应区域却未见有VP IR神经元分布。结论 :VP IR神经元在视交叉上核中的分布有较大的种属差异性     

碱性成纤维细胞生长因子对豚鼠耳蜗螺旋神经节细胞谷氨酸钠毒性损伤的保护作用

目的　在体研究碱性成纤维细胞生长因子 (bFGF)对谷氨酸钠所致豚鼠耳蜗螺旋神经节细胞毒性损伤的拮抗作用。方法　谷氨酸钠组 :谷氨酸钠 2g·kg-1·d-1,ip× 18d ;bFGF组 :bFGF 80 0U·kg-1·d-1,ip ,谷氨酸钠 2g·kg-1·d-1,ip× 18d。停止用药后耳廓反射粗略测定动物听觉水平 ,后取耳蜗 ,固定包埋、切片染色 ,光镜观察。结果　谷氨酸钠组表现为基底转与顶转几乎均匀一致的耳蜗螺旋神经节细胞的消失 ,平均细胞密度为 (1976 3± 10 18 9)个 /mm2 ,占正常的 4 5 % ;bFGF组细胞密度为 (384 0 9± 373 1)个 /mm2 ,约为正常组的 89%。经方差分析 ,F 检验 ,谷氨酸钠组与bFGF组比较 ,耳蜗螺旋神经节存活细胞密度差异显著 ,P <0 0 5。结论　bFGF对豚鼠谷氨酸钠耳蜗旋神经节细胞在体毒性损伤具有保护作用     

MR内耳水成像技术在人工耳蜗植入术中的应用价值

目的 探讨MR内耳水成像技术在人工耳蜗植入术中的应用价值。方法 回顾性分析2015年4月—2016年3月在黑龙江省医院和哈尔滨医科大学附属第四医院行人工耳蜗植入术的128例患者的影像资料,其中男68例、女60例,年龄4～48岁,右耳植入101例、左耳植入27例。患者术前、术后均行头颅内听道螺旋CT及MR内耳水成像检查,并比较两种检测方法对患者内耳、中耳畸形及病变的检出率。结果 MR内耳水成像内听道总体异常检出率90.63%(116/128),CT检出率为76.56%(98/128),二者比较差异有统计学意义(χ²=9.228,P＜0.05)。在前庭导水管异常、耳蜗畸形、内听道狭窄、耳蜗纤维化等疾病,MR诊断的灵敏度、特异度、准确度、阳性预测值和阴性预测值均高于CT,但差异均无统计学意义(P值均>0.05)。结论 在人工耳蜗植入术前、术后进行MR内耳水成像技术检查,可有效了解患者内听道解剖学信息,对治疗方案的选择具有指导性意义,值得临床进一步推广应用。     

儿童蜗神经孔发育不良的低剂量螺旋 CT 诊断简

目的探讨在低剂量CT扫描条件下儿童蜗神经孔发育不良的CT诊断。方法蜗神经孔发育不良患儿35例,其中男性15例,女性20例：年龄3个月～12岁,中位年龄6岁。在低剂量CT扫描的条件下观察35例（48耳）感音性神经耳聋患儿蜗神经孔横径．探讨蜗神经孔发育不良的CT特征。结果诊断蜗神经孔狭窄23例31耳,轴位孔径最大1．4mm,最小O．5mm。双侧8例．单侧15例：其中左侧9例,右侧6例。蜗神经孔封闭12例17耳,双侧5例,单侧7例;其中左侧3例．右侧4例。30耳内听道狭窄,约占63％。结论儿童蜗神经孔发育不良的CT表现为蜗神经孔狭窄或封闭．低剂量螺旋CT扫描是蜗神经孔发育不良的有效诊断方法。     

Bone Marrow Stromal Cells Accelerate Hearing Recovery via Regeneration or Maintenance of Cochlear Fibrocytes in Mouse Spiral Ligaments

Mammalian cochleae have limited capacity for regeneration, which is one of the major difficulties in the treatment of sensorineural hearing loss. In the current study, we examined the potential of bone marrow-derived stromal cells (BMSCs) for functional restoration of mouse cochleae through regeneration or maintenance of cochlear fibrocytes in the spiral ligament (SL). We used a mouse model of degeneration of cochlear fibrocytes in the SL using local application of 3-nitropropionic acid (3-NP), in which disruption of the gap junction network in the SL resulted in the reduction of the endocochlear potential (EP). Mouse BMSCs were infused into the posterior semicircular canal 7 days after 3-NP application. Transplanted BMSCs were frequently observed in the cochlear fluid space 4 weeks after transplantation, although a few transplants had migrated into the cochlear tissues including the SL. BMSC-treated cochleae exhibited higher cell densities in the SL and greater EP levels than the control ones. Immunohistochemistry further demonstrated the restoration of functional proteins in the SL. Significant recovery in thresholds of auditory brainstem responses following BMSC transplantation was found only at 40 kHz in a mild degeneration model. Our cumulative findings indicated that BMSCs accelerated regeneration or maintenance of fibrocytes in damaged SLs, leading to partial functional restoration of the mouse cochleae. Anat Rec, 303:478–486, 2020. © 2019 American Association for Anatomy     

胎儿耳蜗微血管铸型的扫描电镜观察

目的 :观察胎儿耳蜗微血管构筑。方法 :应用微血管铸型技术结合扫描电镜观察 15例胎儿耳蜗血管。结果 :扫描电镜下 ,人类耳蜗血管的三维结构首次得以清晰显示。螺旋蜗轴动脉为供应耳蜗的主要动脉 ,进入蜗轴后反复发出外、中、内放射状动脉。微静脉主要位于鼓阶。结论 :2 8周龄以上胎儿耳蜗血管已发育完善 ,人类耳蜗血管分布呈现明显节段性和区域性。与其生理功能密切相关     

Form and function of the mammalian inner ear

The inner ear of mammals consists of the cochlea, which is involved with the sense of hearing, and the vestibule and three semicircular canals, which are involved with the sense of balance. Although different regions of the inner ear contribute to different functions, the bony chambers and membranous ducts are morphologically continuous. The gross anatomy of the cochlea that has been related to auditory physiologies includes overall size of the structure, including volume and total spiral length, development of internal cochlear structures, including the primary and secondary bony laminae, morphology of the spiral nerve ganglion, and the nature of cochlear coiling, including total number of turns completed by the cochlear canal and the relative diameters of the basal and apical turns. The overall sizes, shapes, and orientations of the semicircular canals are related to sensitivity to head rotations and possibly locomotor behaviors. Intraspecific variation, primarily in the shape and orientation of the semicircular canals, may provide additional clues to help us better understand form and function of the inner ear.     

Detection of immunoreactive atrial and brain natriuretic peptides in the equine atrium

The distribution of immunoreactivity (IR) for cardiodilatin/atrial natriuretic peptide (CDD/ANP) and brain natriuretic peptide (BNP) was examined immunohistochemically and immuno-electron-microscopically in the equine atrium, using specific antibodies. In the immunohistochemical studies, IR-CDD/ANP and IR-pBNP-26 (porcine BNP-26 immunoreactivity) was detected in the cytoplasm of the auricular cardiocytes, but IR-hBNP-32 (human BNP-32 immunoreactivity) was not. The double immunogold labelling method for IR-hBNP-28 and IR-pBNP-26 revealed that gold particles of different sizes were located in the same secretory granules in the cardiocyte, but no gold particles for IR-hBNP-32 were detected. These results show that CDD/ANP and porcine BNP-like peptides are colocalized in the same secretory granules of the equine atrium. They suggest that the equine atrium secretes both CDD/ANP and BNP-like peptides.     

神经生长因子减轻大鼠脑缺血再灌注耳蜗损伤

目的:研究神经生长因子(NGF)对脑缺血再灌注引起的听力及耳蜗损伤的作用。方法:本研究给实验组大鼠脑缺血再灌注前后肌注NGF,对照组动物肌注等量生理盐水,于处理前后对两组的听力损失及耳蜗结构变化加以比较。结果:缺血30min再灌注60min和24h,实验组的听力损失明显小于对照组(P<001)。耳蜗的扫描和透射电镜观察发现,对照组毛细胞静纤毛粘连、部分脱落,外毛细胞肿胀、散在性坏死缺失,螺旋神经节神经元变性,线粒体水肿、嵴断裂。实验组耳蜗Corti’s器毛细胞及螺旋神经节神经元的损伤明显轻于对照组。结论:NGF能减轻脑缺血再灌注引起的听力及耳蜗损伤。     

Immunohistochemical localization of phospholipase C isozymes in mature and developing gerbil cochlea

The possibility that phospholipase C contributes to intracellular signaling in the cochlea was investigated by immunostaining for eight different isoforms of the enzyme. In the mature gerbil cochlea, expression of the isozymes varied widely among different cell types. The phospholipase C-beta1 isoform was detected in inner and outer hair cells, and spiral ganglion neurons where it may participate in regulating Ca(2+) flux. The beta3 isozyme was expressed in epithelial cells thought to mediate lateral and medial circulation of potassium. The beta2 isozyme was present in border, inner phalangeal and Hensen cells, the stria vascularis, and suprastrial and supralimbal fibrocytes where it also may be involved in regulating ion transport activities. The phospholipase C-gamma isozymes were expressed in supporting cells, the stria vascularis, and certain fibrocytes where they possibly participate in activating tyrosine kinase and modulating ion conductances. The delta2 isoform was found in pillar, outer sulcus and strial marginal cells as well as spiral ganglion neurons and their radial processes.Documentation of changes in the expression pattern of phospholipase C isoforms during postnatal development and knowledge of their distribution in several positive control tissues provided further data for speculation about the biologic significance of the cochlear reactivity. The results demonstrate a wide diversity of isozyme distribution in the cochlea and suggest that the enzymes affect activities of various cochlear cell types in different ways.     

Morphogenesis of the Inner Ear at Different Stages of Normal Human Development

This study examined the external morphology and morphometry of the human embryonic inner ear membranous labyrinth and documented its three‐dimensional position in the developing embryo using phase‐contrast X‐ray computed tomography and magnetic resonance imaging. A total of 27 samples between Carnegie stage (CS) 17 and the postembryonic phase during trimester 1 (approximately 6–10 weeks after fertilization) were included. The otic vesicle elongated along the dorso‐ventral axis and differentiated into the end lymphatic appendage and cochlear duct (CD) at CS 17. The spiral course of the CD began at CS18, with anterior and posterior semicircular ducts (SDs) forming prominent circles with a common crus. The spiral course of the CD comprised more than two turns at the postembryonic phase, at which time the height of the CD was evident. A linear increase was observed in the length of anterior, posterior, and lateral SDs, in that order, and the length of the CD increased exponentially over the course of development. Bending in the medial direction was observed between the cochlear and vestibular parts from the latero‐caudal view, with the angle decreasing during development. The position of the inner ear was stable throughout the period of observation on the lateral to ventral side of the rhombencephalon, caudal to the pontine flexure, and adjacent to the auditory ganglia. The plane of the lateral semicircular canal was approximately 8.0°–14.6° with respect to the cranial caudal (z‐)axis, indicating that the orientation of the inner ear changes during growth to adulthood. Anat Rec, 298:2081–2090, 2015. © 2015 Wiley Periodicals, Inc.     

Distinct Localization of Peripheral and Central Types of Choline Acetyltransferase in the Rat Cochlea

We previously discovered a splice variant of choline acetyltransferase (ChAT) mRNA, and designated the variant protein pChAT because of its preferential expression in peripheral neuronal structures. In this study, we examined the immunohistochemical localization of pChAT in rat cochlea and compared the distribution pattern to those of common ChAT (cChAT) and acetylcholinesterase. Some neuronal cell bodies and fibers in the spiral ganglia showed immunoreactivity for pChAT, predominantly the small spiral ganglion cells, indicating outer hair cell type II neurons. In contrast, cChAT- and acetylcholinesterase-positive structures were localized to fibers and not apparent in ganglion cells. After ablation of the cochlear nuclei, many pChAT-positive cochlear nerve fibers became clearly visible, whereas fibers immunopositive for cChAT and acetylcholine esterase disappeared. These results suggested that pChAT and cChAT are localized in different systems of the rat cochlea; pChAT in the afferent and cChAT in the efferent structures.