中耳胆脂瘤上皮增殖能力的研究

目的 :探讨增殖细胞核抗原 (proliferating cell nuclear antigen,PCNA)在中耳胆脂瘤上皮中的表达和定位以及与中耳炎症的关系。方法 :采用 PCNA单克隆抗体对 16例中耳胆脂瘤组织和 6例正常外耳道皮肤 ,面部皮肤以及鼓膜进行免疫定位检测。结果 :PCNA阳性细胞仅见于正常外耳道、面部皮肤和鼓膜的基底细胞层和棘细胞层的开始部位 ,亦可见于胆脂瘤组织上皮全层。上皮下炎性细胞浸润愈重 ,PCNA阳性细胞率愈高。结论 :中耳胆脂瘤上皮组织具有过度增殖能力。上皮下炎症反应可能是角化细胞过度增殖的一个原因     

端粒酶催化亚单位和c-Myc蛋白在中耳胆脂瘤上皮中的表达及意义

目的:观察端粒酶催化亚单位(TERT)和癌基因cMyc的表达产物(cMyc蛋白)在中耳胆脂瘤上皮中的表达情况,分析其在胆脂瘤上皮增殖演变过程中的作用。方法:应用SP法和计算机图像分析系统,检测TERT、cMyc蛋白在32例胆脂瘤组织和14例胆脂瘤患者外耳道皮肤中的表达情况,并进行比较。结果:TERT和cMyc蛋白在所有胆脂瘤上皮各层细胞均存在较强表达,外耳道皮肤表达较弱。各组间胆脂瘤上皮各指标平均阳性细胞率和平均吸收度(平均灰度的倒数),均高于外耳道皮肤,差异有统计学意义(P<0.05);皮下结缔组织伴有大量的炎症细胞,如:淋巴细胞、浆细胞、中性粒细胞和巨噬细胞;胆脂瘤上皮中TERT和cMyc蛋白之间存在正相关关系(P<0.01)。结论:TERT和cMyc蛋白在胆脂瘤上皮的表达从不同方面反映了胆脂瘤上皮的过度增生和角化,二者在胆脂瘤的发病过程中有协同作用;上皮下炎症反应可能是角化细胞过度增殖的一个原因。     

中耳胆脂瘤上皮增殖能力的研究

目的：探讨增殖细胞核抗原（ｐｒｏｌｉｆｅｒａｔｉｎｇ ｃｅｌｌ ｎｕｃｌｅａｒ ａｎｔｉｇｅｎ，ＰＣＮＡ）在中耳胆脂瘤上皮中的表达和定位以及与中耳炎道皮肤关系。方法：采用ＰＣＮＡ单克隆重抗体对１６例中耳胆脂瘤组织例正常外耳道皮肤，面部支及鼓膜进行免疫定位检测。结果：ＰＣＮＡ阳性细胞率愈高。结论：中耳胆脂瘤上皮组织具有过度增殖能力。上皮下炎症反应可能是角化细胞过度增殖的一个原因。     

胆脂瘤上皮过度增殖行为的免疫组化研究

目的 探讨中耳胆脂瘤上皮的过度增殖性和生长方式。方法 用免疫组化SP法观察了表皮生长因子受体（epidermal growth factor receptor,EGFR），增殖细胞核抗原Ki67、Ⅳ型胶原蛋白（collagenⅣ）和层粘连蛋白（laminin,LN）在18例胆脂瘤标本的表达，并与8例外耳道正常皮肤相比较。结果 EGFR有两种染色图案，棕黄色的线状或颗粒状细胞膜强染色和颗粒状的胞浆染色，胆脂瘤标本染色均强于皮肤。在Ki67的免疫染色中，阳性细胞核为棕黄色，胆脂瘤上皮的阳性细胞核比皮肤多。CollagenIV和LN图案极其相似，为连续的宗黄色线状和（或）带状图案。4例胆脂瘤上皮基膜下结膀组织中的血管比皮肤丰富。结论 EGFR和Ki67在胆脂瘤上皮的表达从不同角度反映了胆脂瘤上皮的过度增殖性。CollagenIV和LN的表达说明胆脂瘤属良性病变，血液供应可能比皮肤丰富。     

胆脂瘤上皮过度增殖行为的免疫组化研究

目的　探讨中耳胆脂瘤上皮的过度增殖性和生长方式。方法　用免疫组化SP法观察了表皮生长因子受体 (epidermalgrowthfactorreceptor,EGFR) ,增殖细胞核抗原Ki6 7、Ⅳ型胶原蛋白(collagenIV)和层粘连蛋白 (laminin ,LN)在 18例胆脂瘤标本的表达 ,并与 8例外耳道正常皮肤相比较。结果　EGFR有两种染色图案 ,棕黄色的线状或颗粒状胞膜强染色和颗粒状的胞浆染色 ,胆脂瘤标本染色均强于皮肤。在Ki6 7的免疫染色中 ,阳性细胞核为棕黄色 ,胆脂瘤上皮的阳性细胞核比皮肤多。CollagenIV和LN图案极其相似 ,为连续的棕黄色线状和 (或 )带状图案。 4例胆脂瘤上皮基膜下结缔组织中的血管比皮肤丰富。结论　EGFR和Ki6 7在胆脂瘤上皮的表达从不同角度反映了胆脂瘤上皮的过度增殖性。CollagenIV和LN的表达说明胆脂瘤属良性病变 ,血液供应可能比皮肤丰富     

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。     

应用单克隆抗体Ki-67鉴定中耳胆脂瘤角化细胞的增生性

胆脂瘤是常见的中耳病患，据文献报道胆脂瘤上皮有异常增生活性。Ki67是一种通过细胞表达在细胞增殖周期的民，S，SZ和M阶段能识别人细胞核抗原的单克隆抗体，并能确定标本组织中的细胞增生活性。该作者运用Ki67表达来研究胆脂瘤和正常人皮肤标本，试图证实胆胀瘤上皮的高度增生活性。取鼓室成形术后的胆脂瘤和正常外耳道上皮标本各15份作低温保存，采用活性Ki－67抗体浓度为l：100，运用碱性磷酸酶抗碱性磷酸酶的免疫组化方法。结果显示：局限在正常外耳道上皮基底细胞层的角化细胞，有很少一部分细胞核被Ki67着色，而胆脂瘤上皮基底层…     

HGF在中耳胆脂瘤中的表达和意义

目的 研究肝细胞生长因子(Hepatocyte growth factor,HGF)在中耳胆脂瘤上皮中的表达及与中耳胆脂瘤的血管形成的关系和对胆脂瘤侵蚀行为的影响.方法 采用免疫组织化学SP法检测HGF、微血管计数(Ⅳ型胶原蛋白染色)在34例中耳胆脂瘤组织标本与15例正常外耳道组织中的表达.结果 HGF主要表达于胆脂瘤上皮层和上皮下基质细胞,在正常外耳道皮肤中仅表达于上皮基底层;胆脂瘤基质周围微血管计数均数高于正常外耳道皮肤,二者差异具有显著性(P＜0.05).结论 HGF在中耳胆脂瘤中高表达且与炎症程度呈正相关;微血管计数随着胆脂瘤基质周围炎症程度的增加而增加.HGF诱导的新生血管形成可能是中耳胆脂瘤侵蚀性行为的主要原因之一.     

Caspase-8在中耳胆脂瘤上皮中的表达及意义

目的 研究caspase-8(胱天蛋白酶-8)在中耳胆脂瘤上皮的表达,探讨其在中耳胆脂瘤发病机制中的作用.方法 分别采用免疫组化法和Western Blot(蛋白质印迹法)技术检测caspase-8在36例中耳胆脂瘤组织及20例正常外耳道皮肤组织中的表达情况.结果 caspase-8在36例胆脂瘤上皮各层细胞中均有强表达,而正常外耳道皮肤表达较弱,caspase-8在胆脂瘤上皮中的蛋白表达水平较正常外耳道皮肤显著增高,差异具有统计学意义(P<0.01).结论 胆脂瘤上皮中caspase-8表达较外耳道皮肤显著增高,其可能参与了中耳胆脂瘤上皮细胞的过度凋亡及增殖的调控.     

中耳胆脂瘤上皮过度增殖与MDM2的相关性

目的探讨MDM2（murine double minute）在中耳胆脂瘤上皮增殖机制中的作用,了解胆脂瘤的过度增殖性和生长方式。方法应用免疫组织化学技术检测32例中耳胆脂瘤组和10例正常对照组中MDM2的表达。结果MDM2分布于胆脂瘤上皮全层,但在基底层细胞表达最弱,棘细胞层、颗粒层和角质层依次递增,染色最强的部位是颗粒层和角质层,而正常对照组未见MDM2分布。结论p53／MDM2反馈环异常可能是胆脂瘤上皮异常增殖和凋亡的原因之一。在胆脂瘤基底层细胞表达少量的MDM2可能是维持胆脂瘤不同于恶性肿瘤组织的无限增殖和浸润的基础。     

奥曲肽对鼻咽癌细胞株体外增殖能力的影响

奥曲肽是一种生长抑素类似物,对神经内分泌肿瘤有强大的抑制生长作用,近年来国内外多种研究证实其对多种非神经内分泌恶性肿瘤细胞的生长有抑制作用,而对严重危害我国南方地区的鼻咽癌的作用却研究很少.我们通过体外细胞培养,研究奥曲肽对鼻咽癌细胞株CNE-2增殖的影响.     

Th1: Mediator Lymphocytes in Experimental Autoimmune Labyrinthitis

A previous study on experimental autoimmune labyrinthitis (EAL) consistently demonstrated transient infiltration of lymphocytes only into the inner ear of mice. To clarify the profile of lymphocytes in the initiation of EAL, the present study investigated cell surface antigens, as well as cytokines, from Day 4 to Day 35, using immunohistochemical techniques. Many CD4+ cells mainly infiltrated the endolymphatic sac as early as Day 4 and gradually spread to the rest of the inner ear. Infiltration peaked on Day 12 and persisted in most animals until Day 35, although the number of cells gradually decreased. In contrast, very few CD8+ cells were found to have appeared in the inner ear of all animals on Day 10, and the number of cells rapidly decreased. Many cells positive for IFN- &#110 and IL-2 were identified in the endolymphatic sac on Day 4. These results suggest that helper T1 lymphocytes, rather than cytotoxic T lymphocytes, may play a central role in the initiation of EAL.     

Human Leukocyte Antigen-A,-B,-C and -DR Alleles and Soluble Human Leukocyte Antigen Class I Serum Level in Ménière's Disease

Previous studies have suggested that many human leukocyte antigen (HLA)-A,-B,-C and -DR alleles are associated with Ménière's disease (MD), an inner ear disorder with a proposed autoimmune etiopathogenesis. Despite some discrepancies many reports are in agreement with a hypothesis suggesting an influence of serologically detected HLA-C products in the susceptibility to the disease. To confirm these data we investigated the distribution of HLA-A,-B,-C and-DR antigens that well define the HLA polymorphism using DNA typing. Furthermore, as autoimmune factors have been claimed to play a role in MD, we investigated the serum level of soluble HLA class I (sHLA-I). Molecular typing of HLA class I and II was performed using polymerase chain reaction sequence-specific primers in 41 patients affected by MD, 34 patients affected by other inner ear diseases (OIDs) and 101 healthy subjects. An ELISA technique was employed to investigate the serum level of sHLA-I in 17 MD patients, 10 OID patients and 83 healthy subjects. The results showed a significantly increased frequency of the Cw*07 specificities in MD patients when compared to OID patients (63.4% vs 32.3%; p =6.9 &#50 10 -3 ; relative risk [RR]=3.6) and healthy subjects (63.4% vs 35.6%; p =2.28 &#50 10 -3 ; RR =3.1). The sHLA-I concentrations detected in sera did not differ significantly between MD patients (616 &#45 271 ng/ml), OID patients (570 &#45 307 ng/ml) and healthy subjects (518 &#45 340 ng/ml).     

从皮肤试验分析变应性鼻炎的诊断

目的分析变应性鼻炎的诊断。方法采用皮肤点刺试验检测1248例成人变应性鼻炎患者的变应原。结果517例(41.43%)患者呈阳性皮肤反应,位于前4位的变应原是:屋尘螨、粉尘螨、杂草及树。对1种或2种变应原过敏者占76.4%。对2种变应原过敏的患者,多为具有交叉抗原的变应原。变应原等级为( )以上者占75%。结论皮肤点刺试验是变应性鼻炎诊断的重要手段,可以作出明确的特异性变应原的种类及严重程度的诊断。     

Non-specific immunological determinations in Meniere’s disease: any role in clinical practice?

Several studies supported the hypothesis that an autoimmune response may be a pathogenetic factor in Meniere’s disease occurrence. It has been demonstrated that the endo-lymphatic sac has an important role in the immuno-mediated reaction and it has also become evident that an immunological mechanism may be involved in the development of endo-lymphatic hydrops. The present study was carried out in order to analyse the validity of serological non-specific immune tests in determining the possible role of immune pathology in 200 Meniere’s suffering patients. The immune activity was investigated determining the erythrocyte sedimentation rate (ESR), the C-reactive protein levels, the rheumatoid factor, the serum immunoglobulin levels by group, the complement levels, the lymphocyte sub-populations, the cryoglobulines, the circulating immune complexes (CIC), organ and non-organ auto-antibodies. Levels of Ig, ESR, C-reactive protein, rheumatoid factor, complement fractions and cryoglobulines were either negative or non-significant in the considered cohort of Meniere’s disease suffering patients. On the other hand, in more than half of the sample group high values of CIC were found. Data concerning T-lymphocyte sub-population indicates a clear increase in T-helper compared to T-cytotoxic cells, with an increase in the CD4/CD8 ratio. The datum of particular interest concerns analysis of T-cells activation complexes. Our present findings show that early activated T-lymphocytes change and in particular changes occur in the expression of the interleukin-2 receptor. Moreover, it seems to be possible to correlate the alteration of the immunological tests and the phase of the disease. Present data showed that the immunological factor should be considered a relevant element in most of the cases of Menière’s disease.     

HSP70、Ki-67、nm23-H1蛋白在鼻咽癌组织中表达

目的探讨鼻咽癌组织中HSP70、Ki-67和nm23-H1蛋白表达的临床价值。方法采用免疫组织化学二步法，检测50例鼻咽癌患者鼻咽部或颈淋巴结组织中HSP70、Ki-67和nm23-H1蛋白表达水平。结果50例鼻咽癌组织中HSP70、Ki-67和nm23-H1阳性表达率分别为36．0％（18／50）、98．0％（49／50）和50．0％（25／50）；HSP70和nm23-H1蛋自在Ⅱ—Ⅲ期和Ⅳa期中阳性表达率均有显著性差异（P=0．016，P=0．037）：Binarylogistic回归模型分析HSP70、nm23-H1表达是鼻咽癌局部复发或远处转移的相关因素（P=0．022，P=0．038）：Cox多因素回归模型分析HSP70表达是影响患者生存期的独立因素（P=-0．001）：nm23-H1与HSP70表达呈负相关（P=0．003）；Ki-67表达与鼻咽癌局部复发或远处转移及生存期均无相关性。结论检测鼻咽癌组织中HSP70和nm23-H1表达对指导治疗及评价预后有重要的临床意义。     

喉癌中支原体单克隆抗体与核增殖抗原的表达

目的 探讨支原体感染与核增殖抗原在人喉癌发生、发展及预后中的作用.方法 用免疫组织化学Envision法检测216例喉不同病变组织标本(喉癌145例,喉癌前病变25例,声带息肉31例,癌周喉组织15例)中支原体单克隆抗体PD4蛋白和核增殖抗原Ki67的表达.结果 ①PD4和Ki67在人喉不同病变组织中表达阳性率由高到低分别为喉癌:45.52%(66/145)和82.76%(120/145)、喉癌前病变:16.00%(4/25)和32.00%(8/25)、声带息肉:12.90%(4/31)和22.58%(7/31)、癌周喉组织:6.67%(1/15)和0(0/15),各组间比较有显著性差异(P＜0.01);②中晚期(Ⅲ～Ⅳ期)、伴颈淋巴结转移的喉癌组织中PD4、Ki67阳性表达率均显著高于早期(Ⅰ～Ⅱ期)、无颈淋巴结转移的喉癌组织(P＜0.01).喉癌术后3年或5年存活病例PD4表达阳性率明显低于未存活病例(P＜0.01);而Ki67的表达阳性率在术后3年存活与未存活病例间无统计学差异(P＞0.05),5年存活率间比较有统计学差异为(P＜0.05);③PD4和Ki67在人喉癌组织中联合表达阳性率为32.41%(47/145),显著高于喉其他不同病变组织中联合表达的阳性率(2.81%,2/71,P＜0.01),两者在喉不同病变组织中表达呈正相关(相关系数r＝0.79).结论 PD4和Ki67联合表达与人喉癌的发生、发展及预后有一定的相关性.     

喉鳞状细胞癌组织中人白细胞I类分子和抗原加工递呈分子的表达

目的 研究喉鳞状细胞癌(简称鳞癌)组织中人白细胞抗原(human leucotyte antigen,HLA)Ⅰ类分子和抗原加工递呈(antigen processing machinery,APM)分子抗原转运相关蛋白1(transporter associated with antigen processing,TAP-1)和低分子量蛋白7(low molecular weight polypeptide,LMP-7)下调的频率及其临床意义.方法 用免疫组化方法检测2001-2002年手术切除的51例喉鳞癌组织中HLA-I类抗原重链HLA-ABC和APM分子TAP-1、LMP-7表达情况,并分析其与临床病理特征及预后的关系.结果 HLA-ABC、TAP-1、LMP-7各分子在喉鳞癌组织中的下调率分别是56.9％(29/51)、39.2％(20/51)、45.1％(23/51)；丢失率分别是21.6 ％(11/51)、33.3％(17/51)、27.5％(14/51).HLA-ABC在喉鳞癌组织中的表达分别与TAP-1(r=0.460,P＜0.05)、LMP-7(r=0.685,P＜0.05)的表达呈正相关.在喉鳞癌组织中HLA-ABC、TAP-1、LMP-7的表达均与肿瘤T分期相关(x2值分别为8.61、9.72、8.97,P值均＜0.05),TAP-1、LMP-7的表达分别与肿瘤TNM分期相关(x2值分别为9.18和7.70,P值均＜0.05).单因素分析显示,HLA-ABC阴性表达组与表达下调组、阳性表达组间患者5年累积生存率(cumulative survival rate,CS)及3年无瘤生存率( disease-free survival,DFS)差异有统计学意义(CS组间比较X2值分别为16.18和15.96,DFS组间比较x2值分别为12.54和6.31,P值均＜0.05)；TAP-1阳性表达组与表达下调组、阴性表达组向5年 CS及3年DFS差异有统计学意义(CS组间比较x2值分别为4.23和4.42,DFS组间比较x2值分别为4.12和4.56,P值均＜0.05)；LMP-7阴性表达组与表达下调组、阳性表达组患者5年CS及3年DFS差异有统计学意义(CS组间比较x2值分别为11.46和14.58,DFS组间比较x2值分别为14.17和8.74,P值均＜0.05).多因素分析提示,颈淋巴结转移、复发情况和HLA-ABC表达水平是影响本组喉鳞癌患者预后的重要危险因素(P值均＜ 0.05).结论 喉鳞癌组织中HLA-ABC、TAP-1、LMP-7阴性表达与表达下调不利于肿瘤抗原的递呈及细胞毒性T淋巴细胞向肿瘤实质浸润与杀伤.HLA-Ⅰ类分子、TAP-1、LMP-7阴性表达与表达下调可能促进了癌细胞的发生、侵袭及发展,可能是肿瘤逃避机体免疫监视的方式之一.     

Influence of Human Leukocyte Antigen in the Pathogenesis of Me´nie`re's Disease in the South Korean Population

In order to study the associations of Ménière's disease (MD) with human leukocyte antigen (HLA) alleles in the South Korean population, we examined the distribution of HLA class I and II alleles in 39 MD patients and 199 healthy controls. The frequencies of HLA-Cw*0303 (relative risk (RR) = 2.5; p < 0.02) and -Cw*0602 (RR = 3.7; p < 0.03) were significantly increased in patients with MD compared to controls. However, HLA-B44 (RR = 0.2, p < 0.004) and -Cw*0102 (RR = 0.3; p < 0.03) were significantly decreased in patients compared to controls. When the association between hearing level and the presence of HLA alleles was evaluated, the frequencies of HLA-B13 (RR = 7.4; p < 0.004), -B39 (RR = 45.7; p < 0.003), -Cw*0303 (RR = 4.5; p < 0.02) and -Cw*0602 (RR = 6.5; p < 0.02) were significantly increased and those of -B44 (RR = 0.1; p < 0.02) and -Cw*0102 (RR = 0.1; p < 0.03) were significantly decreased in MD patients with hearing losses (stages II-IV) compared to controls. The frequency of HLA-B13 was significantly higher in MD patients with hearing loss (stages II-IV) than in those with normal hearing (stage I). In terms of HLA-DRB1, -DQA1 and -DQB1 allele frequencies, a significantly higher frequency of DRB1*15 was seen in the patients (RR = 2.4; p < 0.03). These results suggest that some HLA alleles may be a useful genetic marker for predicting prognosis in South Korean patients with MD.