目的探讨重复经颅磁刺激(repetitive Transcranial Magnetic Stimulation,rTMS)对良性阵发性位置性眩晕患者手法复位后残留的头晕症状、步行稳定性及平衡控制能力的临床效果。方法纳入2023年9月至2024年9月期间于空军军医大学第一附属...目的探讨重复经颅磁刺激(repetitive Transcranial Magnetic Stimulation,rTMS)对良性阵发性位置性眩晕患者手法复位后残留的头晕症状、步行稳定性及平衡控制能力的临床效果。方法纳入2023年9月至2024年9月期间于空军军医大学第一附属医院收治的100例耳石症复位后存在残余头晕的患者,按随机区组原则分为干预组与对照组,各50例。两组均给予常规药物基础治疗,干预组加用针对左侧背外侧前额叶区的rTMS治疗,连续5 d;对照组于相同部位接受伪刺激。于干预第6日评估两组眩晕严重程度、步态参数及平衡功能的变化。结果两组患者在性别、年龄、病程、体重指数及合并症等基线资料上均未见显著差异。重复经颅磁刺激干预后,干预组在头晕程度、步态表现及平衡功能的改善均明显优于对照组。治疗过程中,干预组有3例报告短暂轻度耳鸣,均在24 h内自行消失,未予特殊处理;对照组未见不良反应。结论重复经颅磁刺激可安全、有效地缓解耳石症患者复位后的残余头晕,并促进其步态与平衡功能恢复。展开更多
目的分析老年患者听力状况与认知功能的关系,探讨年龄相关性听力损失听觉干预的时机。方法纳入老年受试者266例,采用纯音测听、声导抗测试以及言语分辨率测试进行听力学检查,采用中文版简易精神状态检查量表(mini-mental state examinat...目的分析老年患者听力状况与认知功能的关系,探讨年龄相关性听力损失听觉干预的时机。方法纳入老年受试者266例,采用纯音测听、声导抗测试以及言语分辨率测试进行听力学检查,采用中文版简易精神状态检查量表(mini-mental state examination,MMSE)和数字符号替换测试(digit symbol substitution test,DSST)进行认知功能评估;采用回归分析和受试者工作特征(receiver operating characteristic,ROC)曲线分析探讨言语识别率与纯音听阈的预测价值。结果单因素线性回归分析示,性别、教育水平、重度/极重度听力损失、纯音平均听阈(pure-tone threshold average,PTA)和言语识别率(word recognition score,WRS)与MMSE得分有关。多因素回归分析示,女性、低教育水平以及低WRS的受试者的MMSE得分低于其他群体,高龄、低教育水平和低WRS是DSST得分的危险因素。高教育水平和高WRS是轻度认知障碍(mild cognitive impairment,MCI)的保护因素,在ROC曲线中WRS对MCI的预测能力比PTA更高。ROC曲线分析结果显示,PTA、WRS分别以36.88 dB HL、94%为最佳截断值时,预测MCI的曲线下面积(area under the curve,AUC)分别为0.5919(95%CI:0.518~0.664,P=0.014)、0.630(95%CI:0.558~0.703,P<0.001),灵敏度分别为54.26%、54.26%,特异度分别为61.63%、69.19%;二者联合预测MCI的AUC为0.631(95%CI:0.559~0.703,P<0.001),灵敏度为50.00%,特异度为72.67%。结论言语分辨能力比纯音听阈对MCI有更好的预测能力,当WRS小于94%以及纯音听阈大于36.88 dB HL时可以考虑及时干预,但尚需纵向研究来验证结果。展开更多
The mammalian cochlea relies on outer and inner hair cells(OHCs/IHCs)for sound amplification and signal transmission.Rab3-interacting molecular binding protein 2(RIMBP2),expressed in receptor cells and neurons at syna...The mammalian cochlea relies on outer and inner hair cells(OHCs/IHCs)for sound amplification and signal transmission.Rab3-interacting molecular binding protein 2(RIMBP2),expressed in receptor cells and neurons at synaptic active zones,remains poorly characterized in hearing.We therefore generated a Rimbp2 knockout(KO)mouse model(Rimbp2-/-),which exhibited severe hearing loss with elevated thresholds,prolonged latencies,and reduced amplitudes in auditory brainstem response Wave I.OHC loss via apoptosis was correlated with threshold elevation.In IHCs,patch-clamp recordings revealed reduced exocytosis,including a diminished readily-releasable pool,impaired sustained release,and blocked fast endocytosis.Immunostaining showed unchanged ribbon synapse numbers but positional shifts in the basal pole of KO IHCs.These findings demonstrated RIMBP2’s essential role in OHC survival and its broader regulatory functions in IHC synaptic transmission than previously recognized.展开更多
Cochlear hair cell(HC)damage is a primary cause of sensorineural hearing loss.In this study,we performed metabolomic profiling of cochlear sensory epithelium following neomycin-induced HC injury and identified elevate...Cochlear hair cell(HC)damage is a primary cause of sensorineural hearing loss.In this study,we performed metabolomic profiling of cochlear sensory epithelium following neomycin-induced HC injury and identified elevated arginine metabolism as a key metabolic characteristic of damaged HCs.Using a highly sensitive and specific biosensor,we confirmed that injury induced an increase in arginine levels within cochlear HCs.By manipulating the levels of arginine and its downstream metabolites,we discovered that unmetabolized arginine exerts a strong protective effect on cochlear HCs,independent of its downstream metabolites,such as nitric oxide.Furthermore,integrated metabolomic and transcriptomic analyses revealed that arginine plays a critical role in reprogramming phospholipid metabolism.Arginine supplementation enhanced membrane phospholipid saturation through the Lands cycle and de novo lipogenesis,and protected HCs from phospholipid peroxidation-induced membrane damage and subsequent cell death.Notably,arginine supplementation protected hearing from both noise-and aminoglycoside-induced injury in mice.These findings underscore the role of unmetabolized arginine in modulating phospholipid metabolism and preventing membrane damage in cochlear HCs,highlighting that targeting phospholipid metabolism is an effective hearing protection strategy.展开更多
Purpose Endogenous hydrogen sulfide(H2S)mediates several biological processes and is an important gaseous signal transmitter akin to carbon monoxide or nitric oxide(NO).The role of H2S on auditory signaling path...Purpose Endogenous hydrogen sulfide(H2S)mediates several biological processes and is an important gaseous signal transmitter akin to carbon monoxide or nitric oxide(NO).The role of H2S on auditory signaling pathway remains unknown.We have determined whether ATP can induce generation of H2S in outer hair cells(OHCs)and examined whether H2S can affect the ATP-stimulated generation of NO in OHCs.Methods Individual OHCs were isolated enzymatically from the guinea pig cochlea.Using H2S-and NO-sensitive dyes,6-CdⅡ and 4,5-diaminofluorescein diacetate respectively,we monitored the change in generation of H2S and NO in response to extracellular ATP in OHCs.Results We observed a gradual increase in 6-CdⅡ fluorescence induced by extracellular ATP,providing evidence for H2S production in OHCs.This ATP-induced H2S production was inhibited by suramin,an antagonist of the P2 receptor.ATP did not induce H2S production in a Ca2+-free medium.This suggests that ATP-induced H2S production was mediating by an influx of Ca2+via activation of P2-purinergic receptors.The inhibitor of cystathionine β-synthase(CBS),amino-oxyacetic acid(AOAA),counteracted this increase in 6-CdⅡ fluorescence,which however was unaffected by cystathionine γ-lyase inhibitors,β-cyanoalanine or D-L-propargylglycine.L-NG-nitroarginine methyl ester,a non-selective inhibitor of NO synthase,also failed to inhibit ATP-induced H2S production.In contrast,AOAA inhibited generation of NO in the presence of ATP in OHCs.Conclusion The current study suggests that ATP-induced H2S production is mediating by an influx of Ca2+via activation of P2-purinergic receptors.These findings imply generation of H2S by CBS in response to exposure to ATP in OHCs.Regulatory cross-talk among ATP,H2S,and NO may also exist in the cellular signaling pathways in OHCs.展开更多
Vestibular efferent neurons in the brainstem provide direct cholinergic innervation to peripheral vestibular organs,thereby modulating their sensory responsiveness.In this study,a genetically targeted mouse model with...Vestibular efferent neurons in the brainstem provide direct cholinergic innervation to peripheral vestibular organs,thereby modulating their sensory responsiveness.In this study,a genetically targeted mouse model with choline acetyltransferase-driven fluorescent labeling enabled the precise localization of these neurons to the dorsolateral region of the genu of the facial nerve.Whole-cell patch-clamp recordings in acute brainstem slices revealed that virtually all neurons exhibited spontaneous action potential firing,with marked heterogeneity in discharge patterns and afterhyperpolarization kinetics.Prominent A-type potassium currents were identified and found to be differentially regulated by acetylcholine and calcitonin gene-related peptide.Acute unilateral vestibular deprivation induced a bilateral enhancement of spontaneous firing,indicating sensitivity to altered sensory input.These findings define the intrinsic electrophysiological properties and neuromodulatory mechanisms of vestibular efferent neurons,providing mechanistic insight into their roles in both physiological regulation and adaptive plasticity within the vestibular system.展开更多
目的明确巨噬细胞与噪声导致的隐匿性听力损失发生发展的关系及可能机制。方法通过圆窗注射氯膦酸盐脂质体(LCCA)构建耳蜗巨噬细胞清除小鼠模型,并使用前期本课题组构建的噪声导致的隐匿性听力损失小鼠模型,将小鼠分为对照组(CON组)、...目的明确巨噬细胞与噪声导致的隐匿性听力损失发生发展的关系及可能机制。方法通过圆窗注射氯膦酸盐脂质体(LCCA)构建耳蜗巨噬细胞清除小鼠模型,并使用前期本课题组构建的噪声导致的隐匿性听力损失小鼠模型,将小鼠分为对照组(CON组)、单纯注射LCCA组(LCCA组)、噪声暴露组(NE组)以及噪声暴露伴注射LCCA组(NE+LCCA组)。通过检测小鼠听性脑干反射(ABR)不同频率的听阈、click声刺激听阈,以及80 dB SPL click声刺激诱发的ABRⅠ波振幅及潜伏期来评估各组小鼠听觉功能变化情况;通过耳蜗基底膜铺片染色观察各组小鼠带状突触变化情况;通过定量PCR技术检测各组小鼠促炎因子mRNA水平的表达情况。结果与CON组相比,NE组小鼠的听阈显著升高(P<0.0001),80 dB SPL click声刺激诱发的ABRⅠ波振幅显著降低(P<0.0001)、潜伏期显著延长(P<0.0001),带状突触数目均明显减少(P<0.0001),并伴有内耳促炎因子表达量显著升高(P<0.0001)。NE+LCCA组上述指标较NE组有显著改善(P<0.05),但无法恢复到噪声暴露前状态(P<0.05)。而单纯注射LCCA并不会影响小鼠听觉功能、带状突触数目以及促炎因子表达量。结论巨噬细胞在噪声导致的隐匿性听力损失的发生发展过程中发挥了一定作用,可能的机制是通过加重炎症从而导致内耳的结构和功能受到破坏。展开更多
摘要目的分析老年患者听力状况与认知功能的关系,探讨年龄相关性听力损失听觉干预的时机。方法纳入老年受试者266例,采用纯音测听、声导抗测试以及言语分辨率测试进行听力学检查,采用中文版简易精神状态检查量表(mini-mental state examination,MMSE)和数字符号替换测试(digit symbol substitution test,DSST)进行认知功能评估;采用回归分析和受试者工作特征(receiver operating characteristic,ROC)曲线分析探讨言语识别率与纯音听阈的预测价值。结果单因素线性回归分析示,性别、教育水平、重度/极重度听力损失、纯音平均听阈(pure-tone threshold average,PTA)和言语识别率(word recognition score,WRS)与MMSE得分有关。多因素回归分析示,女性、低教育水平以及低WRS的受试者的MMSE得分低于其他群体,高龄、低教育水平和低WRS是DSST得分的危险因素。高教育水平和高WRS是轻度认知障碍(mild cognitive impairment,MCI)的保护因素,在ROC曲线中WRS对MCI的预测能力比PTA更高。ROC曲线分析结果显示,PTA、WRS分别以36.88 dB HL、94%为最佳截断值时,预测MCI的曲线下面积(area under the curve,AUC)分别为0.5919(95%CI:0.518~0.664,P=0.014)、0.630(95%CI:0.558~0.703,P<0.001),灵敏度分别为54.26%、54.26%,特异度分别为61.63%、69.19%;二者联合预测MCI的AUC为0.631(95%CI:0.559~0.703,P<0.001),灵敏度为50.00%,特异度为72.67%。结论言语分辨能力比纯音听阈对MCI有更好的预测能力,当WRS小于94%以及纯音听阈大于36.88 dB HL时可以考虑及时干预,但尚需纵向研究来验证结果。
基金supported by grants from the National Key R&D Program of China(2021YFA1101300,2021YFA1101800,2020YFA0112503,and 2024YFC2511103)the National Natural Science Foundation of China(82330033,82030029,82401375,81970882,92149304,and 81970883)+7 种基金the Natural Science Foundation of Jiangsu Province(BK20232007)the Science and Technology Department of Sichuan Province(2021YFS0371)Shenzhen Fundamental Research Program(JCYJ20210324125608022)the Open Project Fund of Guangdong Academy of Medical Sciences(YKY-KF202201)Beijing Natural Science Foundation(Z200019)the Jiangsu Provincial Scientific Research Center of Applied Mathematics(BK20233002)the China Postdoctoral Science Foundation(2023TQ0056,2023M730575,and GZC20230435)Jiangsu Funding Program for Excellent Postdoctoral Talent(2023ZB822).
摘要The mammalian cochlea relies on outer and inner hair cells(OHCs/IHCs)for sound amplification and signal transmission.Rab3-interacting molecular binding protein 2(RIMBP2),expressed in receptor cells and neurons at synaptic active zones,remains poorly characterized in hearing.We therefore generated a Rimbp2 knockout(KO)mouse model(Rimbp2-/-),which exhibited severe hearing loss with elevated thresholds,prolonged latencies,and reduced amplitudes in auditory brainstem response Wave I.OHC loss via apoptosis was correlated with threshold elevation.In IHCs,patch-clamp recordings revealed reduced exocytosis,including a diminished readily-releasable pool,impaired sustained release,and blocked fast endocytosis.Immunostaining showed unchanged ribbon synapse numbers but positional shifts in the basal pole of KO IHCs.These findings demonstrated RIMBP2’s essential role in OHC survival and its broader regulatory functions in IHC synaptic transmission than previously recognized.
基金supported by the National Natural Science Foundation of China(82271159,82425018,82071049,81830029,82192860,81922018,82201283,82101219,and 82192861)Shanghai Clinical Medical Research Center for Otolaryngology Diseases(20MC1920200)the STI2030-Major Projects(2022ZD0205400).
摘要Cochlear hair cell(HC)damage is a primary cause of sensorineural hearing loss.In this study,we performed metabolomic profiling of cochlear sensory epithelium following neomycin-induced HC injury and identified elevated arginine metabolism as a key metabolic characteristic of damaged HCs.Using a highly sensitive and specific biosensor,we confirmed that injury induced an increase in arginine levels within cochlear HCs.By manipulating the levels of arginine and its downstream metabolites,we discovered that unmetabolized arginine exerts a strong protective effect on cochlear HCs,independent of its downstream metabolites,such as nitric oxide.Furthermore,integrated metabolomic and transcriptomic analyses revealed that arginine plays a critical role in reprogramming phospholipid metabolism.Arginine supplementation enhanced membrane phospholipid saturation through the Lands cycle and de novo lipogenesis,and protected HCs from phospholipid peroxidation-induced membrane damage and subsequent cell death.Notably,arginine supplementation protected hearing from both noise-and aminoglycoside-induced injury in mice.These findings underscore the role of unmetabolized arginine in modulating phospholipid metabolism and preventing membrane damage in cochlear HCs,highlighting that targeting phospholipid metabolism is an effective hearing protection strategy.
摘要Purpose Endogenous hydrogen sulfide(H2S)mediates several biological processes and is an important gaseous signal transmitter akin to carbon monoxide or nitric oxide(NO).The role of H2S on auditory signaling pathway remains unknown.We have determined whether ATP can induce generation of H2S in outer hair cells(OHCs)and examined whether H2S can affect the ATP-stimulated generation of NO in OHCs.Methods Individual OHCs were isolated enzymatically from the guinea pig cochlea.Using H2S-and NO-sensitive dyes,6-CdⅡ and 4,5-diaminofluorescein diacetate respectively,we monitored the change in generation of H2S and NO in response to extracellular ATP in OHCs.Results We observed a gradual increase in 6-CdⅡ fluorescence induced by extracellular ATP,providing evidence for H2S production in OHCs.This ATP-induced H2S production was inhibited by suramin,an antagonist of the P2 receptor.ATP did not induce H2S production in a Ca2+-free medium.This suggests that ATP-induced H2S production was mediating by an influx of Ca2+via activation of P2-purinergic receptors.The inhibitor of cystathionine β-synthase(CBS),amino-oxyacetic acid(AOAA),counteracted this increase in 6-CdⅡ fluorescence,which however was unaffected by cystathionine γ-lyase inhibitors,β-cyanoalanine or D-L-propargylglycine.L-NG-nitroarginine methyl ester,a non-selective inhibitor of NO synthase,also failed to inhibit ATP-induced H2S production.In contrast,AOAA inhibited generation of NO in the presence of ATP in OHCs.Conclusion The current study suggests that ATP-induced H2S production is mediating by an influx of Ca2+via activation of P2-purinergic receptors.These findings imply generation of H2S by CBS in response to exposure to ATP in OHCs.Regulatory cross-talk among ATP,H2S,and NO may also exist in the cellular signaling pathways in OHCs.
基金National Natural Science Foundation of China(81870724)National Key Research Program Project(2024YFC2418302)+1 种基金Science and Technology Commission of Shanghai Municipality(23DZ2202900)Shanghai Municipal Health Commission Science and Technology Talent Capacity Enhancement Special Project(JKKPYL-2023-A02)。
摘要Vestibular efferent neurons in the brainstem provide direct cholinergic innervation to peripheral vestibular organs,thereby modulating their sensory responsiveness.In this study,a genetically targeted mouse model with choline acetyltransferase-driven fluorescent labeling enabled the precise localization of these neurons to the dorsolateral region of the genu of the facial nerve.Whole-cell patch-clamp recordings in acute brainstem slices revealed that virtually all neurons exhibited spontaneous action potential firing,with marked heterogeneity in discharge patterns and afterhyperpolarization kinetics.Prominent A-type potassium currents were identified and found to be differentially regulated by acetylcholine and calcitonin gene-related peptide.Acute unilateral vestibular deprivation induced a bilateral enhancement of spontaneous firing,indicating sensitivity to altered sensory input.These findings define the intrinsic electrophysiological properties and neuromodulatory mechanisms of vestibular efferent neurons,providing mechanistic insight into their roles in both physiological regulation and adaptive plasticity within the vestibular system.
摘要目的探讨脂代谢指标与噪声下言语识别困难之间的因果关系,并评估其在听力损失早期阶段的作用。方法基于孟德尔随机化(Mendelian randomization,MR)设计,利用英国生物银行和全球脂质遗传学联盟两个大型全基因组关联分析数据库的数据,评估高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇、甘油三酯等脂代谢指标与噪声下言语识别困难之间的因果关联。采用逆方差加权方法作为主要分析方法,并通过多变量MR进一步验证脂代谢指标的独立贡献。结果两样本MR分析表明,HDL-C与噪声下言语识别困难具有显著的保护性因果关系(英国生物银行数据集:OR=0.993,95%CI:0.987~0.999,P=0.014),这一关联在多变量分析中保持稳健(OR=0.992,95%CI:0.984~1.000,P=0.045)。相比之下,其他脂代谢指标未显示出显著的独立因果效应。此外,全球脂质遗传学联盟数据集中HDL-C的保护作用虽然显示出类似趋势,但未达到统计学显著性(OR=0.994,95%CI:0.987~1.000,P=0.054),多变量MR结果(OR=0.992,95%CI:0.983~1.000,P=0.064)。结论HDL-C在噪声下言语识别困难中具有保护作用,HDL-C水平的改善可能通过抑制炎症、抗氧化和改善微循环等机制对听力健康产生积极影响。
摘要目的明确巨噬细胞与噪声导致的隐匿性听力损失发生发展的关系及可能机制。方法通过圆窗注射氯膦酸盐脂质体(LCCA)构建耳蜗巨噬细胞清除小鼠模型,并使用前期本课题组构建的噪声导致的隐匿性听力损失小鼠模型,将小鼠分为对照组(CON组)、单纯注射LCCA组(LCCA组)、噪声暴露组(NE组)以及噪声暴露伴注射LCCA组(NE+LCCA组)。通过检测小鼠听性脑干反射(ABR)不同频率的听阈、click声刺激听阈,以及80 dB SPL click声刺激诱发的ABRⅠ波振幅及潜伏期来评估各组小鼠听觉功能变化情况;通过耳蜗基底膜铺片染色观察各组小鼠带状突触变化情况;通过定量PCR技术检测各组小鼠促炎因子mRNA水平的表达情况。结果与CON组相比,NE组小鼠的听阈显著升高(P<0.0001),80 dB SPL click声刺激诱发的ABRⅠ波振幅显著降低(P<0.0001)、潜伏期显著延长(P<0.0001),带状突触数目均明显减少(P<0.0001),并伴有内耳促炎因子表达量显著升高(P<0.0001)。NE+LCCA组上述指标较NE组有显著改善(P<0.05),但无法恢复到噪声暴露前状态(P<0.05)。而单纯注射LCCA并不会影响小鼠听觉功能、带状突触数目以及促炎因子表达量。结论巨噬细胞在噪声导致的隐匿性听力损失的发生发展过程中发挥了一定作用,可能的机制是通过加重炎症从而导致内耳的结构和功能受到破坏。