【作者】 贾海英；

【导师】 王继群；

【作者基本信息】 暨南大学 ， 眼科学， 2008， 博士

【摘要】 研究鼻眼相关部位解剖和鼻腔鼻窦肿瘤对鼻眼相关外科学具有十分重要的临床价值。目前,这一领域在其相关应用基础学方面鲜见报告。本文就鼻眼显微解剖学、鼻腔鼻窦恶性肿瘤中多基因水平分析等几个部分,从肿瘤的病因、病理和浸润转移等方面进行分子机制研究,通过统计学分析得出一组对临床有指导意义的参数。第一部分:鼻眼相关显微解剖结构研究目的:研究鼻眼相关部位显微解剖结构,探讨其微血管和微淋巴管的分布,为鼻眼相关外科学的基础研究和相关疾病的临床诊断、指导治疗和评估预后提供重要的解剖学依据。方法:取兔的鼻眼相关部位组织经固定、脱钙、脱水、浸腊、包埋、切片后分别行HE染色和免疫组织化学染色;人的标本收集病理科保存石蜡标本。结果:1.13%EDTA中性溶液加微波脱钙所需时间最短,HE染色效果好,组织清晰,免疫组织化学染色效果好;2.兔的眶壁可分成眶上壁、眶前壁、眶下壁和眶后壁,兔最大的鼻窦为上颌窦,兔的鼻眼相关部位于眼眶的前内侧,即眶自前壁;3.在兔及人的鼻眼相关部位标本中CD34、D2-40均可见阳性表达,并且CD34阳性表达明显高于D2-40,说明眼眶与鼻窦交界部位有较多微细血管和少量淋巴管存在。结论:1.13%EDTA中性溶液加微波脱钙法可以作为目前脱钙方法的首选;2.兔有完整的眼眶,可作为眼眶疾病和鼻眼相关疾病研究的动物模型;3.鼻眼相关部位有丰富的血管和少量淋巴管分布,可能在鼻眼相关疾病中发挥的作用。第二部分鼻腔鼻窦恶性肿瘤临床研究目的:研究鼻腔鼻窦恶性肿瘤的流行病学特点和发病规律,为临床的诊断、治疗和评估预后提供依据。方法:回顾性分析137例鼻腔鼻窦恶性肿瘤患者的临床资料和病理资料,将收集的病例用spss统计软件进行统计性描述。结果和结论:(1)病例的发病年龄主要集中在40～76岁年龄段(构成比,105/137);(2)鼻腔鼻窦恶性肿瘤的组织病理学类型非常复杂,上皮源性肿瘤多于软组织肿瘤,鳞癌最常见;(3)鼻腔鼻窦恶性肿瘤患者侵犯部位多样、临床症状复杂。鼻腔恶性肿瘤以鼻塞、涕血为主要临床症状,上颌窦恶性肿瘤以颌面部疼痛、麻木、肿胀和涕血为主,筛窦肿瘤常出现涕血、头痛、眼球突出和视力下降等;(4)内翻性乳头状瘤、横纹肌肉瘤均发生周围组织浸润和侵犯,恶性黑色素瘤和恶性淋巴瘤少见周围组织浸润和侵犯。第三部分p53、p63、p73和nm23-H1与鼻腔鼻窦恶性肿瘤目的:研究抑癌基因p53、p63、p73和nm23-H1与鼻腔鼻窦恶性肿瘤的相关性,探讨其发病的分子学机制,为鼻腔鼻窦恶性肿瘤的病因学提供重要依据。方法:用免疫组化法检测鼻腔鼻窦恶性肿瘤组织、癌旁组织、鼻息肉、内翻性乳头状瘤等组织中p53家族成员p53、p63、p73及nm23-H1等抑癌基因的表达,探讨其与鼻腔鼻窦恶性肿瘤的发生、发展和组织学类型的相关性。结果:1.p53和nm23-H1蛋白在鼻腔鼻窦恶性肿瘤、癌旁和对照组中的表达存在统汁学差异(P＜0.01);2.p53、p63蛋白在鼻腔鼻窦恶性上皮肿瘤和恶性软组织肿瘤中的表达存在统计学差异(P＜0.01);3.p63、p73蛋白在鼻腔鼻窦恶性肿瘤、癌旁和对照组中的表达无统计学差异(P＞0.05);4.p73、nm23-H1蛋白在鼻腔鼻窦恶性上皮肿瘤和软组织肿瘤中的表达无统计学差异,p73蛋白在嗅神经母细胞瘤和恶性黑色素瘤中的表达与对照组的表达均无统计学差异(P＞0.05);5.p53、p63、p73、nm23-H1在有鼻腔鼻窦以外组织侵犯和局限于鼻腔鼻窦的恶性肿瘤中的表达均无统计学差异(P＞0.05)。结论:1.p53和nm23-H1基因在鼻腔鼻窦恶性肿瘤发生中发挥一定作用,p63和p73与鼻腔鼻窦恶性肿瘤的发生无相关性;2.p53和p63基因在不同病理类型的鼻腔鼻窦恶性肿瘤中发挥不同的作用,具有组织特异性,在研究中应根据不同病理类型分类讨论;3.p53、p63、p73和nm23-H1基因在鼻腔鼻窦恶性肿瘤的发展中不发挥作用。第四部分E-cad、LN、LN-R和ColⅣ与鼻腔鼻窦恶性肿瘤目的:研究鼻腔鼻窦恶性肿瘤的浸润性及转移性,为鼻腔鼻窦恶性肿瘤的病因学、诊断学、治疗及评估预后提供重要理论依据。方法:用免疫组化方法检测异腔鼻窦恶性肿瘤组织、癌旁组织、鼻息肉、内翻性乳头状瘤等组织中E-Cad、LN、ColⅣ和LN-R的表达,探讨其与不同组织类型、临床分期的鼻腔鼻窦恶性肿瘤的相关性。结果:1.E-cad、LN、LN-R和ColⅣ蛋白在鼻腔鼻窦恶性肿瘤、癌旁和对照组中的表达存在统计学差异(P＜0.01),LN-R在对照组中鼻息肉或囊肿和内翻性乳头状瘤中的表达也存在统计学差异(P＜0.01);2.E-cad和LN-R蛋白在鼻腔鼻窦恶性上皮肿瘤和恶性软组织肿瘤中的表达存在统计学差异(P＜0.01),LN、ColⅣ在鼻腔鼻窦恶性上皮肿瘤和恶性软组织肿瘤中的表达均无统计学差异(P＞0.05);3.LN-R、LN在不同侵犯范围的鼻腔鼻窦恶性肿瘤中的表达存在统计学差异(P＜0.05),E-cad、ColⅣ在不同侵犯范围的鼻腔鼻窦恶性肿瘤中的表达均无统计学差异(P＞0.05)。结论:1.E-cad、LN、LN-R和ColⅣ在鼻腔鼻窦恶性肿瘤的发生中发挥作用,并且LN-R在鼻内翻性乳头状瘤的发生过程中发挥一定作用;2.E-cad和LN-R在不同病理类型的鼻腔鼻窦恶性肿瘤中发挥不同的作用,具有组织特异性,在研究中应根据不同病理类型分类讨论。而LN和ColⅣ在不同病理类型的鼻腔鼻窦恶性肿瘤中发挥相似的作用;3.LN-R和LN在鼻腔鼻窦恶性肿瘤的发展中发挥一定的作用,与鼻腔鼻窦恶性肿瘤的浸润和侵袭有相关性;而E-cad和ColⅣ在鼻腔鼻窦恶性肿瘤的发展中不发挥重要作用。第五部分CD34和D2-40与鼻腔鼻窦恶性肿瘤目的:研究微血管和微淋巴管在鼻腔鼻窦恶性肿瘤中的作用,为鼻腔鼻窦恶性肿瘤的病因、判断侵袭与转移以及评估预后提供理论依据。方法:用免疫组织化学法检测鼻腔鼻窦恶性肿瘤组织、癌旁组织、鼻息肉和内翻性乳头状瘤中CD34和D2-40的表达,并探讨其与不同组织学类型和不同侵犯程度的鼻腔鼻窦恶性肿瘤的相关性。结果:1.CD34在鼻腔鼻窦恶性肿瘤、癌旁和对照组中的表达存在统计学差异(P＜0.01);2.D2-40在鼻腔鼻窦恶性肿瘤、癌旁和对照组中的表达无统计学差异(P＞0.05),CD34和D2-40在鼻腔鼻窦恶性上皮肿瘤和软组织肿瘤中的表达均无统计学差异(P＞0.05);3.CD34和D2-40在不同侵犯范围的鼻腔鼻窦恶性肿瘤中的表达无统计学差异(P＞0.05);4.用聚类分析法可将MVD计数聚为2类,分别为大于或等于24和小于24两类,经检验得出CD34在鼻腔鼻窦恶性肿瘤、癌旁和对照组中的表达存在统计学差异(P＜0.01),这与MVD计数方差分析结果一致。结论:1.微血管在鼻腔鼻窦恶性肿瘤的发生中发挥重要作用;微淋巴管在鼻腔鼻窦恶性肿瘤的发生中不发挥作用;2.微血管和微淋巴管在不同病理类型的鼻腔鼻窦恶性肿瘤中发挥相似的作用;3.微血管和微淋巴管在鼻腔鼻窦恶性肿瘤的发展中不一定发挥作用,与鼻腔鼻窦恶性肿瘤的浸润和侵袭无相关性;4.认为聚类分析可以用于MVD计数结果聚类,将聚类结果用于鼻腔鼻腔鼻窦肿瘤与CD34的研究,简化了MVD计数步骤,减少了因标本和医生技术差异等因素而导致的误差。在鼻腔鼻窦肿瘤的研究中,MVD计数时可以将24作为等级分界点。更多还原

【Abstract】 It’s very important to research the anatomy of nasocular correlation and malignant tumor in nose.There is no large amount of report for this aspect.There are a large amount of clinical cases in the study on the level of molecules for multiple genes,etiology,prognosis,and outcome in nasal tumors.There are 5 parts in study. There are important parameters in clinical significance by results from statistical analysis.Part 1Objective:(1) To study the micro anatomic structure of the nasocular correlation;(2) To discuss the distribution of the micrangium and micro lymph vessel;(3) To provid important anatomic parameter for the clinical diagnosis,treatment and prognosis of surgery about the nasocular correlation.Methods:The HE and the immunohistchemical technique ware used to detected the fossa orbitalis paries of the rabbit and human.Results:(1) The 13%ethylene diamine tetracetic acid and microwave(EDTA-MW) is the rapiddest decalcification technique and the best staining of the HE and immunohistochemistry;(2) There are four orbital walls on rabbit orbit:paries superior orbitae,paries anterior orbitae,paries inferior orbitae,paries posterior orbitae.The maxillary sinus is the well developed sinus in the rabbit.The paries anterior orbitae is the location of the nasocular correlation;(3) The CD34 and D2-40 protein was expressed in structure of the nasocular correlation in the rabbit,and the positive rate of the CD34 is highter than the D2-40. Conclusion:(1) The 13%EDTA-MW is the first selection of the decalcification techniques;(2) There is a complete fossa orbitalis in rabbit head.The rabbit can be an animal model on studying the disease between the fossa orbitalis and the nasocular correlation;(3) There are redundant blood vessel and small amounts lymph vessel in the correlated structure of nose and eye.Part 2Objective:(1) To study the epidemiological feature and pathogenetic regularity of the nasal and paranasal sinus malignant tumor(NPMT);(2) To provid the important parameter for the clinical diagnosis,treatment and prognosis of the NPMT.Methods:137 patients with NPMT were reviewed retrospectively.Results and Conclusion:(1) Age of onset of the NPMT is constrated between 40 and 76 years;(2) There are many pathological types in MPMT,the disease incidence of the epithelial tissue tumor is higher than the soft connective tissue tumor,the disease incidence of squamous carcinoma is the highest in NPMT;(3) The MPMT encroach many location easily,such as other paranasal sinus,fossa orbitalis,and intracalvarium.The MPMT has complicated clinical symptom;(4) The inverted papillorna and rhabdomyosarcoma of the nasal cavity and paranasal sinus can encroach the circum-structure easily,but the malignant melanoma and malignant lymphoma is contradictory. Part 3Objective:(1) To study the clinical significance of the p53,p63,p73 and nm23H1 genes in NPMT,and discuss the molecular mechanism of the genesis and development in the NPMT;(2) To provid the important parameter for the etiological factor,diagnosis, treatment and prognosis in the NPMT.Methods:The immunohistchemical technique was used to detected the expression of p53、p63、p73 and nm23H1 inNPMT.Results:(1) The positive rate of the p53 and nm23-H1 in NPMT was significantly higher than para-cancer tissues and non-cancer tissues(P＜0.01);(2) There was a significance on the expression of the p53 and p63 in endepidermis and in soft connective tissue of the NPMT(P＜0.01);(3) The expression of p63 and p73 was no significance among the NPMT, para-cancer tissues and non-cancer tissues(P＞0.05);(4) There wasn’t statistical significance on the expression of the p73 and nm23H1 both of endepidermis and in soft connective tissue in the NPMT(P＞0.05).The expression of p73 was not relative to the nasal and paranasal malignant melanoma nor esthesioneuroblastoma(P＞0.05).(5) There was not significance on the expression of the p53,p63,p73 and nm23-H1 in different encroachment degree of the NPMT(P＞0.05)Conclusion:(1) The p53 and nm23-H1 perhaps participate in regulation of occurrenc in NPMT;(2) The p53 and p63 maybe play different role in different pathotype of the NPMT;(3) The expression of p73 may not be associated with tumorgenesis of NPMT.The p53,p63,p73 and nm23-H1 may not be associated with tumorgenesis of NPMT. Part 4Objective:(1) To study the invasion and metastatisis of the NPMT;(2) To provid the important parameter for the etiological factor,diagnosis, treatment and prognosis of the NPMT.Methods:The immunohistchemical technique was used to detected the expression of E-Cad、LN、ColⅣand LN-R in NPMT.Results:(1) There was a significance on the expression of the E-cad、LN、LN-R and ColⅣin NPMT,para-cancer tissues and non-cancer tissues(P＜0.01);and the positive rate of the LN-R in nasal inverting papilloma is significantly higher than that of nasal polyp and cystis(P＜0.01);(2) There was a significance on the expression of the E-cad and LN-R in endepidermis and soft connective tissue of the NPMT(P＜0.01);the expression between LN and ColⅣwas no significance(P＞0.05);(3) There was a significance on the expression of the LN-R and LN in different encroachment degree of the NPMT(P＜0.05);the expression of the E-cad and ColⅣwas not significant(P＞0.05).Conclusion:(1) The E-cad、LN、LN-R and ColⅣperhaps participate in regulation of occurrenc in NPMT,and LN-R perhaps participate in regulation of occurrenc in nasal inverting papilloma;(2) The E-cad and LN-R maybe play different role in different pathotype of the NPMT,but the LN and ColⅣmay be play the homoioplastic role:(3) The expression of LN-R and LN was associated with invasion and metastatisis, and can evaluate the prognoses of the NPSC. Part 5Objective:(1) To study the role of the micrangium and micro lymphangion in NPMT;(2) To provid the important parameter for the etiological factor,diagnosis, treatment and prognosis of the NPMT.Methods:The immunohistchemical technique was used to detected the expression of CD34 and D2-40 in cases of NPSC.Results:(1) The positive rate of the CD34 in NPMT was significantly higher than in para-cancer tissues and non-cancer tissues(P＜0.01),and the expression of D2-40 was no significance(P＞0.05);(2) There wasn’t a significance on the expression of the CD34 and D2-40 in endepidermis and soft connective tissue of the NPMT(P＞0.05),and in different encroachment degree of the NPMT(P＞0.05)(3) The cluster analysis can be used in MVD counted number in NPMT.According to cluster analysis,the MVD counted number can be clustering two group:≥24 and＜24.The analytic result of the cluster is the consonance with MVD counted number.Conclusion:(1) The micrangium perhaps participate in regulation of occurrenc in NPMT,and the micro lymphangion may not be associated with tumorgenesis of NPMT;(2) The micrangium and micro lymphangion maybe play homoioplastic role in different pathotype of the NPMT;(3) The micrangium and micro lymphangion may not be associated with development of NPMT;(4) The cluster analysis can be used in exploratory study about the MVD counted number in NPMT.It can simplify MVD counted number and reduce relative accuracy.更多还原