鼻咽癌中转化生长因子β受体及受体相关蛋白—1的原位表达

目的 研究转化生长因子β（ＴＧＦβ）、其受体及受体相关蛋白－１（ＴＲＩＰ－１）在鼻咽低分化鳞癌中的表达状况。方法 采用免疫组化技术及原位分子杂交技术，检测同一例标本中癌旁上皮和癌细胞内的３型ＴＧＦβ，２种受体的蛋白水平和ＴＲＩＰ－１ｍＲＮＡ水平，分析其变化规律。结果 ＴＧＦβ１、ＴＧＦβ２、ＴＧＦβ３、ＴＧＦβＲＩ、ＴＧＦβⅡ在癌旁上皮表达强于癌组织的百分数分别为６５．７９％、６６．６７％、５５．     

鼻咽癌中转化生长因子β受体及受体相关蛋白-1的原位表达

目的　研究转化生长因子β（ｔｒａｎｓｆｏｒｍｉｎｇ ｇｒｏｗｔｈｆａｃｔｏｒβ，ＴＧＦβ） 、其受体及受体相关蛋白１（ＴＧＦβｒｅｃｅｐｔｏｒｉｎｔｅｒａｃｔｉｎｇ ｐｒｏｔｅｉｎ１，ＴＲＩＰ１） 在鼻咽低分化鳞癌中的表达状况。方法　采用免疫组化技术及原位分子杂交技术，检测同一例标本中癌旁上皮和癌细胞内的３ 型ＴＧＦβ，２ 种受体的蛋白水平和ＴＲＩＰ１ ｍＲＮＡ水平，分析其变化规律。结果　ＴＧＦβ１、ＴＧＦβ２、ＴＧＦβ３、ＴＧＦβＲＩ、ＴＧＦβＲＩＩ在癌旁上皮表达强于癌组织的百分数分别为６５．７９％ 、６６．６７ ％ 、５５．２６％ 、４８．５７％ 、６３．１６ ％ 。癌旁上皮阳性率都高于癌组织阳性率（ Ｐ＜０．０１） ，癌细胞ＴＲＩＰ１ｍＲＮＡ表达平均灰度值１９．３２±１０．７０ ，癌旁上皮平均灰度值为１１．９６ ±５．８５（ Ｐ＜ ０．０５）。结论　鼻咽低分化鳞癌中ＴＧＦβ信息传递通路中的配体、受体和受体相关蛋白１ 表达减弱。     

转化生长因子β_1在改善正常大白鼠与放疗后大白鼠手术切口愈合及皮瓣成活中的作用

头颈部恶性肿瘤患者放疗后再手术常会出现皮肤坏死、切口裂开、瘘管形成等并发症，如何减少和避免上述并发症的发生尚有待研究。用大白鼠研究皮肤慢性放射损伤发生的机理及转化生长因子β1（ＴＧＦ-β１）对改善放疗后再手术切口愈合、皮瓣成活的作用，为临床上有效解决放疗后手术切口问题做了有益的探索。把９５只雄性大白鼠分成４组：第１组，对照组（仅接受手术）；第２组，ＴＧＦ－β１组（接受手术及ＴＧＦ－β1治疗，不接受放疗）；第３组，放疗组（仅接受放疗与手术）；第４组，放疗加ＴＧＦ－β１组（接受放疗、手术及ＴＧＦ－β１…     

头颈鳞癌细胞转化生长因子-β受体表达降低

转化生长因子－β（ＴＧＦ－β）对表达其受体的正常及肿瘤细胞有抗增殖作用，推测ＴＧＦ－β和／或其受体表达降低将导致头颈鳞癌细胞生长失控，该作者对ＴＧＦ－β亚型及其受体亚型在头颈鳞癌中的表达情况进行了研究。应用免疫组织化学方法研究了头颈鳞癌细胞转化生长因子-β及其受体的表达情况，发现７个正常无肿瘤的对照标本均表达ＴＧＦ－β及其受体［（ＴＧＦ－β（ＲＩ）和ＴＧＦ－β（ＲⅡ）］，而４７个头颈鳞癌标本基质表达ＴＧＦ－β，但肿瘤细胞本身不表达ＴＧＦ－β，４０个肿瘤标本不表达ＴＧＦ－β（ＲＩ），４３个肿瘤标本…     

雏鸡听毛细胞损伤后c-myc mRNA及其产物表达的研究

目的：探讨ｃ－ｍｙｃ基因表达在雏鸡基底乳头庆大霉素损伤后听毛细胞再生过程中的意义。方法：应用原位杂交和免疫组化技术检测庆大霉素药后第１、３、７、１４、２１和２８ｄ时ＢＰ损伤区域ｃ－ｍｙｃ ｍＲＮＡ和ｃ－ｍｙｃ蛋白表达情况。结果：ＢＰ修复过程中ｃ－ｍｙｃ、ｍＲＮＡ和ｃ－ｍｙｃ蛋白表达水平明显升高,修复了７ｄ时表达水平达高峰,修复１４ 后ＢＰ中ｃ－ｍｙｃ基因表达水平开始下降。结论：提示ｃ－ｍｙｃ基因表     

β—肌动蛋白在雏鸡毛细胞再生修复过程中的变化

目的 探讨雏鸡基底乳头（basilar papilla ,BP)庆大霉素损伤后不同修复时间听毛细胞β－肌动蛋白（β－actin）和β－肌动蛋白（β－actin mRNA）的变化。方法 选10d龄依沙褐种鸡48只,36只给予庆大霉素（每天100mg/kg体重）肌肉注射,连续10d,于停药后第1、3、7、14、21和28天各处死动物6只,另12只用作同期正常对照,每组2只。分别应用免疫组化和原位杂交技术检测BP损伤区域β－actin和βactin mRNA表达状况,并对免疫组化和原位杂交染色进行计算图像分析。结果 在停药后的1、3、7d,BP中β－actin免疫反应物呈现由阴性逐渐转为阳性反应的变化,至第14天时,其阳性反应物强度接近或达到正常水平,以后增强缓慢。停药后第7天,BP听毛细胞β－actin mRNA先于β－actin表达至高峰。结论 鸡BP损伤后14d新生的毛细胞静纤毛已分化成熟,可能为其功能恢复提供了物质基础。     

GDNF转基因表达对庆大霉素所致的耳蜗和前庭毒性的影响

神经胶质细胞源性神经营养因子（ｇｌｉａ ｃｅｌｌ ｌｉｎｅ－ｄｅｒｉｖｅｄ ｎｅｕｒｏｔｒｏｐｈｉｃ ｆａｃｔｏｒ ＧＤＮＦ）是转化生长因子－β超家族的一个远亲成员,在发育和成熟的耳蜗中均起潜在作用。为了解ＧＤＮＦ转基因表达对庆大霉素所致的耳蜗和前庭毒性的影响,该文作者将３２只豚鼠分为６组：（１）人工外淋巴组（４只）;（２）Ａｄ．ＬａｃＺ组（４只）;（３）庆大霉素组（４只）;（４）Ａｄ．ＬａｃＺ＋庆大霉素组（６只）;（５）Ａｄ．ＧＤＮＦ＋庆大霉素组（救援组,８只）;（６）应用庆大霉素后７天＋Ａｄ．Ｇ…     

喉鳞状细胞表皮生长因子受体mRNA的表达及其意义

目的：了解喉部鳞状细胞表皮生长因子受体（ＥＧＦ－Ｒ）ｍＲＮＡ的表达情况及其与喉部鳞癌的特征，组织分化的关系。方法；用斑点杂交法检测２０例喉鳞状细胞癌活体标本。结果：１５例出现ＥＧＦ－Ｒ ｍＲＮＡ表达，其表达率为７５Ａ％。ＥＧＦ－Ｒ ｍＲＮＡ表达与患者年龄，肿瘤的部位无关；而与ＴＮＭ分期，组织分化程度有关，Ｔ３，Ｔ４期的表达高于Ｔ１，Ｔ２期；分化程度低的肿瘤，其ＥＧＦ－Ｒ ｍＲＮＡ表达高于分化程度中     

转化生长因子β受体的生物学特征及其与头颈肿瘤关系研究现状

转化生长因子β（ＴＧＦ－β）是多效能因子,对上皮细胞起负性生长调节作用。其生物学效应的发挥均由其相应的受体介导,以ＴＧＦ－βⅠ型、Ⅱ型受体尤为重要。近几年发现肿瘤细胞、转化细胞失去对ＴＧＦ－β负性生长调节作用的敏感性,与ＴＧＦ－β受体的缺失、不表达或突变密切相关。本文在受体水平上就ＴＧＦ－β受体与头颈肿瘤的关系做一综述。     

雏鸡基底乳头Fas蛋白表达与细胞凋亡的关系

目的 :研究耳毒性药物致雏鸡内耳损伤过程中细胞凋亡与Fas蛋白表达的关系。方法 :应用卡那霉素 10 0mg/ (kg·d)皮下注射 ,连续 10d ,建立雏鸡内耳损伤模型。在雏鸡内耳损伤后 1、3、7、14、2 1d采用免疫组化方法、TUNEL法 ,分别对基底乳头Fas蛋白免疫反应阳性细胞和凋亡细胞分布进行观察。结果 :TUNEL阳性细胞主要分布在雏鸡基底乳头腔面毛细胞层 ,较高水平标记发生在停药后第 7天 ,停药后第 14天 ,TUNEL阳性细胞开始减少。Fas蛋白表达从停药后第 1天开始增高 ,停药后第 3天表达水平达高峰 ,随后逐渐下降。结论 :Fas蛋白在雏鸡基底乳头损伤后的高水平表达与毛细胞凋亡紧密相关 ,可能参与了损伤过程中的毛细胞凋亡调控。     

小鸡再生听毛细胞及其突触连接的放射自显影

目的 采用光镜放射自显影（Ｌｉｇｈｔ ｍｉｃｒｏｓｃｏｐｉｃ ａｕｔｏｒａｄｉｏｇｒａｐｈｙ，ＬＭ－ＡＲＧ）及电镜放射自显像（Ｅｌｅｃｔｒｏｎ ｍｉｃｒｏｓｃｏｐｉｃ ａｕｔｏｒａｉｏｇｒａｐｈｙ，ＥＭ－ＡＲＧ）技术，观察不鸡再生听毛细胞及其突触连接的特征。方法 ７ｄ龄纯种伊莎鸡２０只，实验组（ｎ＝１４），连续皮下注射庆大霉素１０ｄ；对照组（ｎ＝６）。皮下注射生理盐水，用药第４ｄ开始注射氚村记胸腺     

Beta-actin changes during hair cell regeneration in the chick basilar papilla]

OBJECTIVE: To investigate the changes of beta-actin and beta-actin mRNA expression during hair cell regeneration in the chick basilar papilla(BP) following gentamicin ototoxicity. METHODS: Thirty-six chicks were given muscular injection of gentamicin at dosage of 100 mg/kg.d for 10 consecutive days, and twelve additional chicks received injection of normal saline instead of gentamicin as controls. At 1, 3, 7, 14, 21 and 28 days after the treatment, six chicks in experimental group and two chicks in control group were sacrificed. Basilar papillas were prepared for immunohistochemistry and in situ hybridization study. beta-actin and beta-actin mRNA expression in the damaged region were histologically analyzed by computer image system. RESULTS: beta-actin immunoreactivity of BP changed significantly after treatment with gentamicin. From the first day to the fourteenth day after gentamincin, beta-actin immunoreactivity turned positive gradually. It approximated the normal response level at the fourteenth day after treatment of gentamincin. Then increasing rate of beta-actin immunoreactivity positive response level slowed from the fourteenth day after the treatment. At seventh day after treatment, beta-actin mRNA expression in BP increased to peak value, which preceded the peak expression of beta-actin in the regenerated hair cells. CONCLUSION: Nascent hair cells seem to mature 14 days after hair cell damage, which may play a role in auditory functional recovery.     

庆大霉素和卡那霉素耳中毒后雏鸡耳蜗毛细胞的再生

给２大龄纯种罗丝鸡分别肌注庆大霉素（ＧＭ）和卡那霉素（ＫＭ）１０天，于停药当天、第１、２、３和４周处死小鸡，取出内耳基底乳头（ＢＰ）用扫描电镜和光镜观察，发现从ＢＰ的基底到中部有广泛性损害，毛细胞损失率约占４０％，在１周后受损部位表皮板上出现丛生的有微绒毛的再生毛细胞，随恢复期延长，再生毛细胞表面积增大，动纤毛和静纤毛来长出，３至４周受损部位已完全被发育成熟的再生毛细胞填补，ＢＰ形态恢复。     

Secretion of a new basal layer of tectorial membrane following gentamicin-induced hair cell loss

Scanning electron microscopy (SEM) and video-enhanced DIC light microscopy were used to assess morphological changes in the chick tectorial membrane (TM) following gentamicin-induced hair cell loss. Gentamicin was administered (100 mg/kg/day for 3 days) and isolated and in-situ TMs were examined in both fixed and unfixed preparations at days 5 and 10 after the initial injection. Although this protocol induced hair cell damage extending up to 75% of the length of the basilar papilla, there was no apparent damage to the TM itself. However, the ejection of damaged hair cells appeared to sever the filamentous attachments between the TM and the apical surface of the basilar papilla. In SEM preparations this detachment caused the TM to shrink back toward the superior edge. Interestingly, despite the lack of TM damage, gentamicin treatment did reveal the secretion of a new basal layer of TM. Secretion of this new basal layer had begun by day 5 and it was well organized by day 10. This new layer formed attachments to both the recovering basilar papilla and the overlying original TM, a step thought to be necessary for the restoration of auditory function in the regenerating cochlea.     

Regeneration of the tectorial membrane in the chick cochlea following severe acoustic trauma

Damage to the tectorial membrane caused by acoustic trauma was examined with scanning and transmission electron microscopy immediately after exposure and at selected time points over a 10 day recovery period. At 0 h of recovery the structure of the tectorial membrane overlying the region of hair cell damage was severely disrupted and connections between the membrane and the basilar papilla were lost. By 24 h of recovery, regeneration of the tectorial membrane was evident in the secretion of new matrix materials by the supporting cells of the basilar papilla. By 10 days of recovery a new honeycomb-like matrix had replaced the segment of damaged tectorial membrane, re-established connections with hair cell stereocilia and become fused with adjacent regions of undamaged tectorial membrane. However, the regenerated segment included only the honeycomb-like structure of the lower layer of the normal tectorial membrane. The laterally-oriented fibers which form the upper layer of the membrane were not regenerated over the damaged region. These findings indicate that the tectorial membrane is regenerated in parallel with the hair cells during recovery from acoustic trauma but the full extent of this recovery and its effect on cochlear function are not yet clear.     

高强纯音声损伤鸡耳蜗后毛细胞再生研究

本文报告给2天日龄雏鸡每日1.5kHz120dBSPL高强纯音暴露8小时，共刺激6天；随后分0.1、2和3周处死动物，其耳蜗基底乳头（Bp)行扫描电镜（SEM）和光镜（LM）观察,结果发现1.5kHz强纯音仅造成鸡Bp基部至中部内侧较窄范围毛细胞损害，毛细胞消失率占30％左右，当恢复1周后，受损处毛细胞迅速再生，至恢复2周后Bp基本正常。证明高纯度纯音损害鸡内耳后受损区毛细胞有再生能力，再生毛细胞发育成熟而使Bp恢复正常形态。     

Hair cell damage and recovery following chronic application of kanamycin in the chick cochlea

Three-day old chicks were given kanamycin at a dose of 200 mg/kg/day for 10 days and their cochleae were processed for scanning electron microscopy at 1, 3, 7 and 14 days following the last injection. Both hair cells and supporting cells were damaged by kanamycin in the basal 35% of the basilar papilla. By 14 days post-treatment, however, most of the damaged region had been replaced with regenerating hair cells and supporting cells. The base-to-apex gradient of morphological development along the cochlea was observed in the process of regeneration. Kinocilium and microvilli were observed on the apical surfaces of the regenerating hair cells.     

Hair cell regeneration in the chick basilar papilla after exposure to wide-band noise: evidence for ganglion cell involvement

It has been demonstrated that the auditory epithelium in the chick basilar papilla may regenerate after acoustic or ototoxic damage. Both types of damage may elicit the appearance of new cells that may develop in to the sensory cells. Factors inducing this process and the role of ganglion cells, the first neuron cells in the auditory pathway, are still unknown. The pattern of auditory damage and regeneration, after octave-band and pure-tone noise exposure, has been well established in research studies on chicks, but there are scarce data on wide-band noise effects. The aim of this study was to investigate the effect of wide-band noise, with different exposure levels applied, on the chick basilar papilla and supporting cells. Further, it was also aimed to determine whether the proliferation of ganglion cells, after wide-band noise exposure, occurs. The morphological changes were assessed with fluorescent, light, and transmission electron microscopy. Cell proliferation was studied based on immunoreactivity assays of proliferating cell nuclear antigen (PCNA). The exposure to wide-band noise at 120 dB SPL for 72 h produced stripe-like lesion of tall hair cells along the neural edge of the basilar papilla, mainly in the middle and, at the lesser extend, in its proximal part. There was no patch-like damage to the region of short hair cells, commonly observed after the exposure to the octave-band or pure-tone noise. The lesion extend depended on the level of exposure. The lower equivalent levels of noise (120 dB SPL for 40 h intermittent exposure) produced proportionally less damage. No morphological changes at light and fluorescent microscopy (apart from tectorial membrane exfoliation) were observed at 110 dB SPL in case of 20 h intermittent exposure. The elimination of dying hair cells took place either by pulling a damaged cell down to the basilar membrane or by extruding the cell to the subtectorial space. New hair cells reappeared at the sensory epithelium on the fifth day after the end of exposure. Cell proliferation started prior to hair cell loss. PCNA-like immunoreactivity was observed after the exposure at all levels in both the damaged and intact areas. PCNA appeared not only in the supporting cells, as indicated in previous studies, but also in the ganglion cells, suggesting ganglion cell involvement in the process of regeneration.     

Hair cell regeneration and recovery of auditory thresholds following aminoglycoside ototoxicity in Bengalese finches

Birds regenerate auditory hair cells when original hair cells are lost. Regenerated hair cells become innervated and restore hearing function. Functional recovery during hair cell regeneration is particularly interesting in animals that depend on hearing for vocal communication. Bengalese finches are songbirds that depend on auditory feedback for normal song learning and maintenance. We examined the structural and functional recovery of the Bengalese finch basilar papilla after aminoglycoside ototoxicity. Birds were treated with the ototoxic aminoglycoside, amikacin, daily for 1 week. Treatment resulted in hair cell loss across the basal half of the basilar papilla and corresponding high frequency hearing loss. Hair cell regeneration and recovery of auditory brainstem responses were compared in the same animals. Survival times following treatment were between 1 day and 12 weeks. Analysis of structural recovery at weekly intervals indicated that hair cells in the Bengalese finch papilla require a maximum of 1 week to regenerate and appear with immature morphology at the epithelial surface. An additional 6 days are required for adult-like morphology to develop. Repopulation of the damaged region was complete by 8 weeks. Recovery of auditory thresholds began 1 week after treatment and reached asymptote by 4 weeks. Slight residual threshold shifts at 2.0 kHz and above were observed up to 12 weeks after treatment. Direct comparison of structural and functional recovery indicates that auditory thresholds recover maximally before a full complement of hair cells has regenerated.