黏附分子及血管内皮生长因子在鼻息肉中的表达

目的 探讨细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）以及血管内皮生长因子（VEGF）在鼻息肉中的表达与意义。方法25例鼻息肉标本和9例下鼻甲黏膜标本，分别行ICAM-1、VCAM-1、VEGF免疫组化染色和HE染色，光镜下观察比较各种分子表达水平。结果 鼻息肉中可见大量嗜酸性粒细胞（EOS）浸润。ICAM-1、VCAM-1和VEGF均可表达于鼻息肉血管内皮、间质及炎症细胞，且在鼻息肉血管内皮、间质及浸润炎症细胞中的表达趋势呈现正相关关系。结论 ICAM-1、VCAM-1可能与EOS等炎症细胞附壁浸润活化过程关系密切，VEGF可能启动并加强这一病理变化。它们的协同作用可能参与了鼻息肉的病理过程。     

黏附分子在实验性变应性鼻炎鼻黏膜中的表达

变应性鼻炎迟发相反应阶段是以嗜酸粒细胞为代表的大量的炎性细胞的浸润而导致的慢性炎症，血管细胞黏附分子-1(vascular cell adhesion molecule-1，VCAM-1)、细胞间黏附分子-1(intercellular adhesion molecule-1，ICAM-1)在嗜酸粒细胞等炎性细胞的黏附、浸润到黏膜炎症区域起着重要作用。本研究观察实验性变应性鼻炎鼻黏膜中黏附分子的表达。     

细胞因子和细胞黏附分子在慢性鼻窦炎中的表达及意义

目的探讨细胞因子和细胞黏附分子在慢性鼻窦炎发病机制中的作用.方法应用免疫组化SABC法检测两型慢性鼻窦炎患者上颌窦黏膜内白细胞介素-1(IL-1)、IL-8、干扰素-γ(IFN-γ)和粒细胞巨噬细胞集落刺激因子(GM-CSF)以及细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1).结果在慢性单纯性鼻窦炎Ⅰ型组中IL-1,IL-8阳性细胞数明显高于对照组(P<0.01),而Ⅱ型组中IL-8,IFN-γ和GM-CSF表达明显高于对照组(P<0.01),Ⅰ型组IL-1明显高于Ⅱ型组(P<0.01).ICAM-1的表达两组鼻窦炎无明显差异,而明显高于对照组(P<0.01),VCAM-1各组表达均较少,无明显差异.结论细胞因子及ICAM-1在慢性鼻窦炎发病中起一定的作用.通过对不同细胞因子类型及黏附分子的研究可进一步了解慢性鼻窦炎的发病机制.     

嗅神经离断后大鼠嗅黏膜气味受体的表达变化

明显减少,差异具有统计学意义(t=63.960,P=0.000).随着术后恢复时间的延长,嗅神经离断侧再生嗅黏膜表达Olr1493基因受体的细胞逐渐增多,至第6周时基本和对照侧一致,分别为(417.8±32.4)个和(445.3±10.0)个,差异无统计学意义(t=2.600,P=0.060).结论 大鼠嗅神经离断后再生嗅感觉神经元及其受体数量可以恢复至正常水平,表达位置亦未发生变化.     

细胞间黏附分子-1和白细胞介素-6在变应性鼻炎鼻和支气管黏膜中的表达及意义

目的 :探讨细胞间黏附分子 1(ICAM 1)、白细胞介素 6 (IL 6 )在变应性鼻炎鼻及支气管黏膜中的表达及意义。方法 :采用免疫组织化学方法检测ICAM 1、IL 6在 2 8例变应性鼻炎患者 (鼻炎组 )及 12例健康志愿者 (对照组 )的鼻、支气管黏膜中的表达。结果 :ICAM 1在鼻炎组鼻和支气管黏膜中的表达率分别为 10 0 .0 0 %和92 .86 % ,其差异无统计学意义 (P >0 .0 5 ) ;对照组鼻、支气管黏膜中ICAM 1表达率 (分别为 6 6 .6 7%和 5 8.33% )分别与鼻炎组 (分别为 10 0 .0 0 %和 92 .86 % )比较 ,差异均有统计学意义 (分别P <0 .0 1和P <0 .0 5 )。IL 6在鼻炎组鼻、支气管黏膜中的表达率分别为 10 0 .0 0 %、82 .14 % ,其差异无统计学意义 (P >0 .0 5 ) ;对照组鼻、支气管黏膜中IL 6表达率均为 4 1.6 7% ,分别与鼻炎组 (分别为 10 0 .0 0 %和 82 .14 % )比较 ,差异均有统计学意义 (分别P<0 .0 1和P <0 .0 5 )。结论 :ICAM 1、IL 6是呼吸道变态反应性炎症中的重要细胞因子 ,上、下呼吸道变应性炎症密切相关 ;同时治疗上、下呼吸道炎症性疾病 ,才能取得更好的治疗效果。     

细胞黏附分子及一氧化氮合酶在变应性鼻炎鼻黏膜中的表达

目的:对细胞黏附分子(CAM)及一氧化氮合酶(NOS)进行原位观察,探讨它们在变应性鼻炎(AR)发病中的作用。方法:采用链霉卵白素-生物素复合体(SABC)法,对AR患者及对照组手术切除的下鼻甲黏膜内细胞间黏附分子-1(ICAM-1)、血管CAM-1(VCAM-1)和淋巴细胞功能相关抗原-1(LFA-1),以及神经型NOS(nNOS)、诱导型NOS(iNOS)和内皮细胞型NOS(eNOS)进行原位检测。结果:AR下鼻甲黏膜内3种CAM表达的阳性细胞数ICAM-1为[(14.4±2.2)个/HP(×400),以下同],LFA-1为(17.2±3.3)个/HP,VCAM-1为(11.5±2.7)个/HP;对照组下鼻甲黏膜内3种CAM表达的阳性细胞数ICAM-1为(8.7±1.8)个/HP,LFA-1为(10.3±2.1)个/HP,VCAM-1为(6.9±1.8)个/HP。t值分别是11.57,10.02和8.07(均P<0.01)。AR及对照组下鼻甲黏膜内nNOS表达的阳性细胞数分别为(9.4±1.7)个/HP和(4.7±1.3)个/HP,t值为12.62,(P<0.01);iNOS表达的阳性细胞数分别为(27.5±3.2)个/HP和(4.3±1.7)个/HP,t值为36.03(P<0.01)。eNOS表达的阳性细胞数分别为(6.5±2.1)个/HP,(6.2±1.9)个/HP,t值为0.62(P>0.05)。结论:CAM在黏膜上皮、腺上皮、血管内皮以及黏膜下的各种炎性细胞等的表达,说明CAM参与AR的发生、发展。nNOS和iNOS在AR的发病过程中可能起重要作用。     

人鼻黏膜上皮细胞中白细胞介素12和细胞间黏附分子1及单核细胞趋化因子3的表达研究

目的:利用RT-PCR检测原代培养的人鼻黏膜上皮细胞中IL-12、细胞间黏附分子1(ICAM-1)以及单核细胞趋化因子3(MCP-3)mRNA的表达。方法:获取人鼻黏膜组织,原代培养人鼻黏膜上皮细胞。设计IL-12 p35亚基、IL-12 p40亚基I、CAM-1以及MCP-3的引物,参照物采用3-磷酸甘油醛脱氢酶(GAPDH),目标扩增片段938 bp。半定量RT-PCR检测其mRNA在鼻黏膜上皮细胞的表达。结果:×400光镜下观察结果显示原代培养的鼻黏膜上皮细胞形状呈圆形或不规则形,饱满贴壁。RT-PCR结果通过1%琼脂糖凝胶电泳显示,鼻黏膜上皮细胞有IL-12 p35I、CAM-1以及MCP-3 mRNA的表达,但无IL-12 p40亚基的表达。结论:人鼻黏膜上皮细胞中有IL-12 p35、ICAM-1以及MCP-3的mRNA表达,但无IL-12p40亚基的表达。人鼻黏膜上皮细胞不能合成完整有活性的IL-12。     

嗅感觉神经细胞的分离与培养

目的 建立哺乳动物嗅感觉神经细胞体外培养的方法。方法 取成年家兔的嗅区粘膜，用胰酶和胶原酶消化法进行嗅感觉神经细胞的分离与原代培养，并应用抗神经微丝蛋白(NFP)抗体、抗嗅觉标记蛋白(OMP)抗体进行免疫细胞化学染色，同时采用甲苯胺蓝神经特异性染色和扫描电镜对其进行鉴定。结果 从嗅粘膜分离以及原代培养得到的嗅感觉神经元，光镜、电镜下为典型的双极神经元，OMP、NFP阳性，神经元特异性染色阳性。分离的嗅感觉神经元可在体外存活数小时，其细胞形态、生存数量和存活时间均能满足体外实验的基本要求。结论 本实验在家兔建立了较稳定、可靠的嗅感觉神经细胞分离和原代培养方法。为深入开展嗅感觉神经细胞的离体研究，提供了较为理想的材料来源和技术保障。     

可溶性细胞黏附分子－1与喉癌侵袭转移的关系

细胞间黏附分子-1（intercellutar adhesion molecule,ICAM-1）在肿瘤侵袭转移过程中起重要作用,可溶性ICAM-1（soluble ICAM-1,SICAM-1）是细胞膜型ICAM-1经水解脱落而成,其抗原性与ICAM-1相同。本研究通过测定喉癌患者血清中SICAM-1,探讨SICAM-1在喉癌侵袭转移中的生物学作用及临床意义。     

老年突发性聋患者血液纤维蛋白原和黏附分子测定及其临床意义

目的　探讨老年突发性聋患者血液纤维蛋白原,可溶性细胞间黏附因子（soluable intercellular adhesion molecule-1,sICAM-1）,可溶性血管内皮黏附因子-1（soluable vascular cell adhesion molecule-1,sVCAM-1）的变化以及不同分型突发性聋患者的差异。方法　40例老年突发性聋患者按照中华医学会最新的诊断治疗指南分型,并按有无合并症进行分组,40例健康的老年体检者作为对照组。测定血液纤维蛋白原水平,sICAM-1和sVCAM-1表达,结果进行统计学分析。结果　老年突发性聋血纤维蛋白原、sICAM-1和sVCAM-1表达均高于对照组,差别有显著性意义。不同突发性聋分型患者间以上指标的表达无显著性差别,但有合并症突发性聋以上指标的表达高于无合并症,差别有显著性意义。结论　老年突发性聋血液纤维蛋白原、sICAM-1和sVCAM-1表达增高,微循环障碍可能在老年突发性聋发病中占主导地位。     

Upregulation of neural growth-associated protein and neural cell adhesion molecule in mouse olfactory epithelium and axons after unilateral removal of the olfactory bulb

The expression of synaptosomal-associated protein (SNAP-25), neural growth-associated protein (GAP-43) and neural cell adhesion molecule (NCAM) were studied in mouse olfactory cells and axons for 2 weeks following unilateral bulbectomy. The olfactory cells and axons in the control olfactory epithelium were positive for SNAP-25 but levels decreased in the atrophic olfactory epithelium 3 days after bulbectomy. There was no expression of SNAP-25 in the olfactory epithelium on the bulbectomy side 7 days after bulbectomy, indicating that this protein may be a good marker for the degeneration of olfactory cells. The expression of NCAM was still found in the atrophic olfactory epithelium at 7 days after bulbectomy, while the expression of NCAM in the olfactory epithelium of the bulbectomy side was stronger than that on the control side at 14 days after bulbectomy. The expression of GAP-43 in the olfactory axonal bundles of the bulbectomy side at 3 and 4 days after bulbectomy was stronger than that on the control side. These results suggest that upregulation of NCAM may be related to the regeneration of the olfactory cells, with upregulation of GAP-43 probably playing a role in axonal regeneration after bulbectomy. Received: 6 January 1998 / Accepted: 22 April 1998     

Maintenance of regional difference in cellular composition of neurospheres derived from adult mouse olfactory bulb

Neurospheres are potentially good tools to study olfactory neurogenesis. For this purpose, it is necessary to characterize neurospheres derived from the olfactory system. This study is aimed at identifying the regional difference of cellular composition within neurospheres derived from the adult mouse olfactory bulb, and studying whether these characteristics are maintained throughout the long-term culture. Neural cells were obtained from the olfactory bulbs of 8-week-old Balb/c mice and the dissociated cells were cultured in serum-free media containing epidermal growth factor and basic fibroblast growth factor to form neurospheres. These neurospheres were subjected to characterization by confocal microscopy after immunofluorescence staining with nestin, proliferating cell nuclear antigen (PCNA), neuronal cell adhesion molecule (NCAM), glial fibrillary acidic protein (GFAP), O4, and olfactory marker protein (OMP). Confocal microscopy showed that nestin-reactive cells, which are stem cell-like cells, were present at the periphery of neurospheres and PCNA-reactive cells undergoing proliferation were found throughout the neurospheres. Neuronal cell markers, NCAM-reactive cells, were observed at the center of neurospheres while glial cell markers, GFAP- or O4-reactive cells, were present at the periphery of neurospheres. No cells were found to be reactive for OMP, a differentiated olfactory neuron marker. This regional distribution of cells in neurospheres was maintained through 17 passages for a year. The neural lineage of cells showed different distribution within neurospheres and this localization was preserved throughout the culture period. These findings in the neurosphere culture system of the olfactory bulb may help to study olfactory neurogenesis.     

人嗅黏膜细胞色素P4502A表达的免疫印迹和免疫组化分析

目的　研究人类细胞色素P4 5 0 (cytochromeP 4 5 0 ,CYP 4 5 0 ) 2A(CYP2A)在鼻黏膜中的表达特点和意义。方法　应用免疫印迹方法分别检测 36例鼻腔鼻窦疾病患者和孕 96、119、16 0、182、185d的 5例胎儿不同部位鼻组织中CYP2A的表达水平 ,并用免疫组化方法检测CYP2A在嗅区黏膜中分布特点。结果　成人标本中 10块嗅区黏膜标本中 8例免疫印迹阳性 ,而 37块呼吸区黏膜标本均为阴性。胎儿嗅区黏膜标本免疫印迹均为阳性 ,其表达水平随胎龄有增加的趋势 ,仅 1例呼吸区黏膜检测到CYP2A。免疫印迹定量分析显示 ,成人嗅区鼻黏膜微粒体CYP2A蛋白质水平在0 1～ 1 1pmol/mg之间 ,胎儿的鼻微粒体蛋白质含量在 0 8～ 5 3pmol/mg之间 ,胎儿一般高于成人。免疫组化结果显示 ,CYP2A免疫活性存在于嗅黏膜的支持细胞和Bowman腺。结论　CYP2A存在于嗅黏膜提示 :嗅黏膜是吸入性外源性物质生物转化的靶位 ,CYP2A还与嗅觉感受有关 ,胎儿期CYP2A嗅区鼻黏膜的表达也可能使母体衍生毒素对胎儿成长发育有潜在威胁     

大鼠嗅球神经干细胞的超微结构

目的研究体外培养的大鼠嗅球神经干细胞(neuralstemcell,NSC)的超微结构。方法采用添加丝裂原的无血清培养基分离、培养出生后第1天大鼠嗅球NSC,分别于培养后1周,1个月及2个月收集NSC,2.5%戊二醛固定,行扫描电镜及透射电镜观察。结果体外培养的NSC呈圆形或椭圆形,表面有微突起或微绒毛,细胞核浆比例高,稀少的细胞浆中有多少不等的未成熟线粒体、核糖体及高尔基复合体。不同培养时期的神经球内未分化NSC的结构大致相同,均有分裂增殖细胞及少数凋亡细胞。培养1～2个月后出现存在粗面内质网及粗长突起的分化细胞。相邻分化细胞的突起间有缝隙连接。结论体外培养的大鼠嗅球NSC保持原始细胞的形态和结构,并能分化成神经细胞。     

Neural cell adhesion molecule expression in adenoid cystic carcinoma of the head and neck

OBJECTIVE: To investigate whether there is a correlation between neural cell adhesion molecule (NCAM) expression and perineural spread in patients with adenoid cystic carcinoma of the head and neck (ACCHN). STUDY DESIGN: Retrospective review of medical records and immunohistochemical staining of specimens from 37 patients treated at the University of Arkansas in Little Rock from 1987 to 1997. METHODS: Sections from paraffin-embedded specimens were examined for the presence of NCAM using monoclonal anti-NCAM antibody by avidin-biotin-peroxidase immunohistochemical staining. NCAM staining was scored in each specimen and correlated with the data obtained from patient charts. RESULTS: Twenty-five of 37 specimens (68%) showed histopathological evidence of perineural spread. All 37 specimens (100%) stained positive for NCAM, regardless of perineural spread status. CONCLUSION: Our results suggested that the use of NCAM expression as a predictor of perineural spread is highly unlikely.     

大鼠嗅上皮神经干细胞的分离及培养

目的：从新生及成年大鼠嗅上皮分离培养嗅上皮神经干细胞（NSC）,研究其增殖及分化特性。方法：采用含10％胎牛血清的DMEM／F12（1：1）培养液,分离、培养生后第3天和硫酸锌原位创伤后6d的成年大鼠嗅上皮NSC,应用间接免疫荧光染色鉴定培养的NSC及分化的特异性神经细胞类型,四甲基偶氮唑盐法（MTT）测定嗅上皮NSC的生长曲线及生长因子的影响。结果：从生后第3天和成年大鼠嗅上皮分离、培养出巢蛋白免疫反应阳性而细胞角蛋白免疫反应阴性,并能分化为神经元和星形胶质细胞的NSC。生后第3天和成年大鼠嗅上皮NSC生存能力的差异无统计学意义（P〉O．05）,细胞克隆形成率为0．05％～0．10％。碱性成纤维细胞生长因子可以显著促进嗅上皮NSC的增殖。结论：从生后第3天和成年大鼠嗅上皮可培养出具有自我复制和多向分化潜能的NSC。     

Examination and classification of human olfactory mucosa in patients with clinical olfactory disturbances

Summary To determine objectively the degree of olfactory disturbance, we biopsied the olfactory mucosa from patients who complained of anosmia. The olfactory disturbances in this study were caused by choanal atresia, chronic sinusitis, viral inflammation, and head trauma, as well as by congenital and idiopathic anosmia. The biopsy specimens were examined by light microscopy and the degree of mucosal degeneration present was classified according to five grades. The clinical courses of the patients studied paralleled the changes found in the olfactory mucosa.     

Abnormalities of axon growth in human olfactory mucosa

OBJECTIVES/HYPOTHESIS: Random biopsies of the human adult olfactory mucosa often demonstrate degenerative changes in the olfactory epithelium (OE) in both dysosmic and normosmic patients and, consequently, have limited diagnostic usefulness. However, detailed analysis of the subepithelial tissue with specific attention to the fascicles of the olfactory nerve and abnormalities of axonal growth may improve the correlation of histopathology with sensory function. STUDY DESIGN: Retrospective review of human OE biopsies. METHODS: Mucosal biopsies from the olfactory area obtained from 27 subjects were examined by light and electron microscopy, with particular attention to the olfactory nerve fascicles; results were correlated with clinical status. Immunohistochemical analysis was used to characterize the extent of axonal depletion, relative maturity of the parent population, and aberrant axonal growth. RESULTS: As expected, there are areas of respiratory metaplasia and neuronal depletion in normosmic as well as dysosmic patients. The degree of axon degeneration within the fascicles correlates better with individual olfactory status. Immature neurons predominate, and re-entrant neuromas develop in patients with olfactory loss caused by disconnection from the olfactory bulb. Individuals with olfactory loss caused by epithelial damage as with chronic rhinosinusitis display evidence of nerve fascicle degeneration and intraepithelial neuromas. CONCLUSION: The status of olfactory axons provides useful information on the overall condition of the olfactory periphery and improves the diagnostic usefulness of mucosal biopsies. In addition to an assessment of the epithelium per se, the fascicles of the olfactory nerve need to be characterized for a complete analysis of the olfactory mucosa.     

神经特异性烯醇化酶及嗅标记蛋白在不同胎龄胎儿嗅黏膜中的表达

目的　探讨神经特异性烯醇化酶 (neuron specificenolase ,NSE)及嗅标记蛋白 (olfactorymarkerprotein ,OMP)在不同胎龄胎儿嗅黏膜中的表达。方法　以免疫组化方法检测NSE及OMP在1 2、1 6、2 0、2 4、2 8和 34周 6例不同胎龄胎儿嗅黏膜中的表达。结果　NSE免疫阳性反应在孕 1 2～ 34周的胎儿嗅黏膜中均有表达 ,各胎龄胎儿嗅黏膜切片中均可见大量阳性着色的双极嗅神经细胞。孕1 2周时 ,阳性细胞数量很多 ,排列紧密 ,胞体多位于嗅上皮中下部且阳性嗅神经上皮占据胎儿鼻腔上2 /3的黏膜 ,随着孕龄的增大 ,阳性细胞逐渐成多层次排列 ,所占鼻腔黏膜面积却逐渐减小 ,至孕 34周时 ,仅局限于鼻腔上 1 /3黏膜。OMP免疫反应在孕 1 2周胎儿鼻腔上 2 /3黏膜切片中仅发现少量阳性着色的双极神经细胞 ,明显少于同龄胎儿NSE阳性反应细胞。随着胎龄的增加 ,OMP阳性细胞逐渐增多 ,且胞体多位于嗅上皮中上部 ,但其数量仍相对少于同龄NSE阳性细胞。结论　人胚胎 1 2周时嗅黏膜中已有大量的嗅神经细胞 ,其中少数嗅神经细胞已开始发育成熟。随着胎龄的增大 ,嗅上皮面积逐渐缩小 ,但成熟的嗅神经细胞逐渐增多 ,胚胎发育后期的胎儿嗅化学感受器发育已趋于成熟