表皮生长因子及其受体在中耳慢性鼓膜穿孔病变中的作用

目的　探讨表皮生长因子及其受体在中耳慢性鼓膜穿孔病变中的作用。方法在豚鼠慢性创伤性鼓膜穿孔的动物模型上 ,观察应用表皮生长因子治疗的鼓膜愈合率。在具有典型鼓膜后皱襞处穿孔的胆脂瘤型慢性化脓性中耳炎患者的病理标本上 ,应用免疫组化SP染色方法和计算机图像分析系统 ,检测 10例鼓膜穿孔部位邻近皮肤表皮生长因子受体的表达情况。结果　在动物实验中表皮生长因子治疗组鼓膜愈合率明显高于对照组 (χ2 =9 92 3,P <0 0 1)。未发现有诱发中耳胆脂瘤的病例。在临床研究中鼓膜穿孔邻近皮肤表皮生长因子受体的阳性率为 (39 3± 7 4 ) % ( x±s,下同 ) ,耳道深部正常皮肤为 (2 5 4± 3 7) %。 2组间差异具有高度显著性意义。结论　表皮生长因子有明显促进鼓膜穿孔愈合的能力。表皮生长因子受体在鼓膜穿孔部位邻近皮肤的中等表达 ,表明使用表皮生长因子治疗有一定的理论基础和有诱发中耳胆脂瘤的风险。     

表皮生长因子受体在胆脂瘤的异常表达

为了研究表皮生长因子与胆脂瘤之间的关系，利用免疫组化ＳＬＡＢ法，对１８例中耳胆脂瘤和８例外耳道正常皮肤的ＥＧＦ受体进行了检测。结果显示ＥＧＦ受体在胆脂瘤的表达强于皮肤，表明在胆脂瘤内有ＥＧＦ受体的存在和调节异常，进一步证实了胆脂瘤上皮的过度增殖性，同时也表明胆脂瘤的过度增殖与ＥＧＦ受体有关。     

表皮生长因子受体在胆脂瘤的异常表达

为了研究表皮生长因子（ｅｐｉｄｅｒｍａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＥＧＦ）与胆脂瘤之间的关系，利用免疫组化ＳＬＡＢ法，对１８例中耳胆脂瘤和８例外耳道正常皮肤的ＥＧＦ受体进行了检测。结果显示ＥＧＦ受体在胆脂瘤的表达强于皮肤，表明在胆脂瘤内有ＥＧＦ受体的存在和调节异常，进一步证实了胆脂瘤上皮的过度增殖性，同时也表明胆脂瘤的过度增殖与ＥＧＦ受体有关。     

表皮生长因子受体和Ki67及p16在成人中耳胆脂瘤中的表达

目的:研究表皮生长因子受体(EGFR)、Ki67、p16在成人中耳继发性胆脂瘤上皮中的表达情况,分析它们之间的相互关系,探讨其表达对胆脂瘤上皮侵袭能力的影响。方法:应用免疫组织化学SP染色方法检测EGFR、Ki67和p16在30例成人中耳胆脂瘤上皮、21例成人胆脂瘤患者外耳道正常皮肤、17例正常人外耳道中的表达情况,应用计算机图像分析系统对其阳性表达进行定量分析。结果:EGFR、Ki67、p16在成人中耳继发性胆脂瘤上皮中阳性表达率分别为70.0%,60.0%,46.7%,与外耳道正常皮肤相比表达均差异有统计学意义。成人中耳胆脂瘤中EGFR、Ki67与p16之间表达均无相关性(均P>0.05)。胆脂瘤侵袭能力与EGFR、Ki67表达有显著相关性(均P<0.01)。EGFR、Ki67表达灰度值越低,表达密度越高,胆脂瘤侵袭能力越强。p16在成人中耳胆脂瘤中的表达与侵袭能力之间无相关性(P>0.05)。EGFR、Ki67、p16在成人中耳胆脂瘤中阳性细胞主要分布于上皮全层,以基底层和棘层为著,呈高度表达;而在对照组中阳性细胞仅在基底层表达,呈弱表达。结论:EGFR、Ki67、p16在成人中耳胆脂瘤中呈高表达,EGFR、Ki67的表达与成人中耳胆脂瘤的侵袭能力有高度相关性,提示成人中耳胆脂瘤具有高度增殖能力,其中细胞因子EGFR、Ki67、p16起到重要的作用。     

角化细胞生长因子在中耳胆脂瘤的表达及意义

目的研究角化细胞生长因子（keratinocyte growth factor,KGF）在中耳胆脂瘤中的表达,探讨KGF对胆脂瘤上皮增殖能力的影响及其在中耳胆脂瘤发生、发展中的作用。方法20例胆脂瘤上皮标本及配对的正常外耳道皮肤标本制成石蜡切片,应用免疫组化SP染色法检测上述两组标本中KGF和Ki67的表达,对染色结果进行定量分析。结果KGF在胆脂瘤上皮中呈强阳性表达,且从基底层向角质层染色有逐渐增强的趋势,间质中可见散在的阳性细胞;正常外耳道皮肤标本主要表现为间质中稀疏不均的弱阳性表达,在上皮细胞不表达。胆脂瘤上皮和正常外耳道皮肤的KGF阳性表达率之间差异有显著性意义（P〈0.01）。在胆脂瘤上皮中KGF与Ki67表达呈正相关（r=0.609,P〈0.01）。结论KGF和Ki67表达与中耳胆脂瘤的增殖能力有高度相关性,局部炎症通过调控KGF的表达促进胆脂瘤上皮的异常增殖,其中KGF的自分泌机制可能与胆脂瘤的发生、发展密切相关。     

表皮生长因子及其受体在中耳慢性鼓膜穿孔病变中的作用

OBJECTIVE: To evaluate the possible roles of epidermal growth factor(EGF) and its receptor (EGFR) on the chronic tympanic membrane perforations. METHODS: A phosphate buffer saline of EGFR was administered to a Gelfoam pledget placed over chronic tympanic membrane perforations in guinea pigs. The EGFR of 10 specimens from the acquired middle ear cholesteatoma of the adjacent skin around perforation was examined by immunohistochemical SP method and computer image analysis. Results Complete closure of the tympanic membrane perforations was observed in 82.6% of EGF-treated ears, but only 33.3% in the controls(P < 0. 01). No case was led to middle ear cholesteatoma in the experiment group (0/23). The positive expression in the adjacent skin around perforation was (39.3 -/+ 7.4)%; and the normal external ear skin was (25.4 +/- 3.7)%; There were distinctly significant differences between the adjacent skin around perforation and the normal external ear skin (P < 0. 01). CONCLUSIONS: EGF is effective on closing chronic tympanic membrane perforations in the guinea pigs. Present data suggests that EGF-treated may induce the occurrence of middle ear cholesteatoma.     

表皮生长因子受体在鼻咽癌及癌前病变组织中的表达

运用免疫组织化学标记链霉菌素－生物素技术，检测表皮生长因子受体在正常备咽粘膜、鼻咽粘膜异型增生和鼻咽癌中的表达。发现鼻咽正常粘膜全部为阴性，异型增生阳性表达率较低，而一旦鼻咽粘膜癌变后，表达率明显升高。ＥＧＦＲ的表达与鼻咽癌的低分化性相关，ＥＧＦＲ表达的肿瘤易发生淋巴结转移，５年生存率较低。提示ＥＧＦＲ的表达与鼻咽癌的发生有一定关系，并可作为判断鼻咽癌生物行为和预后的一个有用指标。     

中耳胆脂瘤上皮PTEN基因的表达及意义

目的探讨PTEN基因在中耳胆脂瘤形成机制中的作用。方法应用免疫组化染色和逆转录聚合酶链反应(RT-PCR)方法检测32例中耳胆脂瘤组织和10例正常外耳道皮肤组织中PTEN的表达情况。结果中耳胆脂瘤组织和正常外耳道皮肤组织均有PTENmRNA和蛋白的表达,且两组间无统计学差异(P>0.05)。结论在胆脂瘤形成过程中,PTEN的作用更趋向于维持上皮细胞的正常生理状态,而不是发生基因异常导致细胞非正常增殖。     

喉癌表皮生长因子受体表达的免疫组化定量分析

运用免疫组化方法检测了喉癌、喉粘膜不典型增生、声带从组织各１８便中表皮生长因子受全（ＧＦＲ）的表达水平，结合显微图像分析仪进行定量分析，结果表明：在声带息肉组织中ＥＧＦＲ存在于基底细胞层，偶见于棘细胞层，而在喉粘膜不典型增生及喉癌组织的各层细胞均有表达。喉癌细胞中ＥＧＦＲ含量明显高于声带息肉及喉粘膜不典型增生细胞（均为Ｐ〈０．０１），病理分级不同的喉癌细胞ＥＧＦＲ含量不同，Ⅱ、Ⅲ级鳞状细胞癌中均较     

中耳胆脂瘤肥大细胞形态学观察

对３０例中耳胆脂瘤肥大细胞进行组织化学与透射电镜观察，并与健康外耳道皮肤对照．结果发现中耳胆脂瘤肥大细胞明显多于正常对照组（Ｐ＜０．０１），主要分布范围包括上皮层、下皮下特别是近骨吸收面的肉芽组织内。电镜下大多数肥大细胞呈活化状态，与ＴＣ肥大细胞类似，呈无定型均质细颗粒状或毛玻璃样改变，肥大细胞贴壁的小静脉内见许多多形核白细胞溢出；活化肥大细胞主要呈“缓慢脱颗粒”状态；少数肥大细胞与成纤维细胞及浆细胞密切接触。讨论了肥大细胞在中耳胆脂瘤发病和发展中的可能作用。     

Expression of EGFR and Microvessel Density in Middle Ear Cholesteatoma

Objectives

Cholesteatoma destructs bony tissue by the interactions between hyperproliferative epithelial cells and subepithelial inflammatory cells. The aim of this study was to evaluate the expression of epidermal growth factor receptor (EGFR) and microvessel density (MVD) in middle ear cholesteatoma tissue in an effort to determine the relationship between expression of EGFR and neovascularization.

Methods

We evaluated the expression of EGFR and MVD by immunohistochemical staining for CD31 and Factor VIII in 32 cholesteatoma tissue samples and 7 normal postauricular skin samples. We also analyzed the correlation between EGFR expression and MVD.

Results

The expression of EGFR was higher in cholesteatoma than in postauricular skin, but the difference was not statistically significant. EGFR was more highly expressed in the suprabasal layer than in the basal layer. Using CD31 and Factor VIII, we analyzed the MVD and found that it was significantly higher in cholesteatoma than in postauricular skin, and significantly correlated with the expression of EGFR.

Conclusion

Our results suggest that overexpression of EGFR and neovascularization are correlated with the growth of cholesteatoma.     

MMP-9和EGFR在中耳胆脂瘤中的表达及作用

目的探讨基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)和表皮生长因子受体(epi-dermal growth factor receptor,EGFR)在中耳胆脂瘤骨质破坏中的作用及相互关系。方法采用免疫组织化学技术和计算机图像定量分析法,检测MMP-9、EGFR在22例中耳胆脂瘤上皮组织、10例中耳肉芽组织及22例外耳道深部正常皮肤中的表达,分析二者在中耳胆脂瘤上皮组织的表达与骨质破坏程度的相互关系。结果 MMP-9、EGFR均表达于中耳胆脂瘤上皮组织、中耳肉芽组织及外耳道深部正常皮肤,但中耳胆脂瘤上皮组织中二者的含量显著高于中耳肉芽组织及外耳道深部正常皮肤,MMP-9在中耳胆脂瘤上皮组织中的表达高于EGFR,二者的表达具有相关性,且与骨质破坏的程度有关,骨质破坏范围越广,二者的表达越高(P<0.05)。结论 MMP-9和EGFR在中耳胆脂瘤组织中的表达与中耳胆脂瘤的骨质破坏密切相关,且二者的表达与骨质破坏程度呈正比,二者之间可能存在协同作用;MMP-9在中耳胆脂瘤的骨质破坏中可能具有重要作用。     

M CP -1和 FN 在中耳胆脂瘤上皮中的表达和意义

目的探讨单核细胞趋化因子（monocyte chemotactic factor ,MCP -1）和纤维连接蛋白（fibronectin , FN）在继发性中耳胆脂瘤上皮中的表达及其对胆脂瘤上皮侵袭能力的影响。方法应用免疫组织化学MaxVi-sionTM法检测MCP-1和FN在30例中耳胆脂瘤上皮、20例胆脂瘤患者耳后正常皮肤、16例非胆脂瘤患者耳后正常皮肤中的表达,应用计算机图像分析系统对其阳性表达灰度值情况及分析,比较三组之间 MCP -1和 FN表达的差异。结果 MCP-1阳性细胞表达主要分布于胆脂瘤上皮全层,其中棘层呈高表达,MCP-1在中耳胆脂瘤上皮中阳性表达率为70％,灰度值为147．2±20．1,强于胆脂瘤患者耳后正常皮肤中的阳性表达（35％,200．8±18．4）和非胆脂瘤患者耳后正常皮肤中的阳性表达（37．5％,193．3±15．5）（ P＜0．05）。FN阳性细胞主要分布于胆脂瘤上皮全层,基底层、棘层和基质呈高表达,FN在中耳胆脂瘤上皮中阳性表达率为76．7％,灰度值为139．2±18．5,强于胆脂瘤患者耳后正常皮肤中的阳性表达（30％,195．0±12．9）和非胆脂瘤患者耳后正常皮肤中的阳性表达（31．3％,191．6±13．5）（P＜0．05）。在30例中耳胆脂瘤上皮组织中,MCP-1和FN的灰度值与胆脂瘤的侵袭能力负相关（rmcp-1＝-0．682,rfn ＝-0．531,P＜0．01）,MCP-1和FN蛋白的表达不存在线性相关。结论 MCP-1和FN均在成人中耳胆脂瘤中高表达,且与中耳胆脂瘤的侵袭能力呈负相关。     

The role of middle ear effusions and epidermal growth factor in cholesteatoma formation in the gerbilline temporal bone

To study the process of aural cholesteatoma formation, we used gerbilline temporal bones to examine histologically the early stages of spontaneous cholesteatomas associated with experimentally induced otitis media with effusion (OME) following electric cauterizations of the eustachian tube. Epidermal growth factor (EGF) was then localized immunohistochemically in the pars flaccida of normal ears and the forming spontaneous cholesteatomas. Findings in the ears with the early spontaneous cholesteatomas were effusion inside the pars flaccida and hypertrophy and hyperkeratosis of the pars flaccida. Findings in the ears with experimental OME involved an effusion in the whole middle ear cavity as well as hypertrophy and hyperkeratosis in both the pars flaccida and pars tensa. The incidence of ear drum changes was higher in the experimental OME group than in control animals without cauterization. EGF was localized in the mucous layer of normal drums, the mucous layer and lamina propria of drums with hypertrophy alone, and all lalers in drums with hypertrophy and hyperkeratosis. EGF was especially positive in the cytoplasms of transformed cuboidal cells. These findings suggest that EGF within the transformed mucous layer may play an important role as a biochemical factor in developing cholesteatomas.     

分泌性中耳炎的中耳共振频率变化

对正常人６０耳及分泌性中耳炎８４耳，进行中耳共振频率和鼓室导抗图测试，显示正常人组中耳共振频率为８１４±１６４Ｈｚ，中耳积液组中耳共振频率为２９２±１３４Ｈｚ，中耳积液时，质量增大，要达到共振，共振频率必须向低频区移动。治愈后１～２个月，有３８耳再进行多频率扫描，中耳共振频率为７８６±１３９Ｈｚ，与正常人组接近。分泌性中耳炎８４耳中，鼓室导抗图分别为：Ｂ型６９耳、Ｃ型１０耳、Ａ型５耳；而８４耳中中耳共振频率有７９耳＜４２６Ｈｚ（积液组Ｘ＋Ｓ）。在临床上，中耳共振频率的改变、Ｂ型鼓室导抗图，结合鼓膜改变，能更迅速、更准确地诊断本病     

实验性中耳积液大鼠的鼓室导抗图测试

目的 建立客观、稳定的检测大鼠中耳积液的方法。方法 将健康SD大鼠15只（30耳）建立实验性中耳职液大鼠模型，利用GSI－33Version Ⅱ型中耳分析仪，加用自制探头，分别对建立模型前后的大鼠进行鼓室导抗图检测，获取相关指标。结果 正常大鼠鼓室导抗图测试图型为A型或As型，大鼠中耳积液模型的鼓室导抗图测试有26耳变为B型，4耳仍为有峰型图但其导纳值、峰压值和梯度值均发生相应变化。造模前后大鼠鼓室导抗图曲线平滑，A型和As型对称性较好。结论 鼓室导抗图测试能够稳定、客观、敏感地反映大鼠的中耳传音功能。     

Expression of epidermal growth factor, epidermal growth factor receptor and transforming growth factor-alpha in the human fetal inner ear

The expression of epidermal growth factor (EGF), EGF receptor and transforming growth factor (TGF)-alpha was analyzed in the human fetal inner ear using immuno-histochemical methods. EGF receptor was observed only in 9.5-week-old fetal vestibular epithelia. In 14- and 16-week-old fetuses, EGF receptor could not be detected. TGF-alpha was observed strongly in the 9- and 11-week-old vestibular epithelia, whereas only trace amounts were detectable in the 14- and 16-week-old vestibular epithelia. These findings suggest that EGF and TGF-alpha probably have a mitogenic effect in the sensory epithelia of the fetal inner ear, especially at early stages of development.