Abstract研究背景： 耳廓假性囊肿是耳鼻喉科的常见病、多发病。其病因以及发病机制尚未完全阐明，该病在亚洲人群发病明显多于欧美。现国内外多数学者认为其发病与以下因素有关：外伤及局部机械刺激；胚胎及遗传学说；溶酶体学说；免疫因素；软骨膜学说；蛋白聚糖学说。由于该病病因不清，治疗方法多样，但易复发，常导致耳廓增厚、变形。 耳廓假性囊肿的组织病理学改变尚存在争议，有学者认为耳廓假性囊肿形成是由于LDH的释放引起耳廓软骨软化及液化；也有学者认为耳廓假性囊肿囊壁是由软骨变性、退化、坏死所引起；亦有学者通过对耳廓假性囊肿外壁进行病理学观察，发现囊肿外壁的软骨膜是形成囊肿外壁的基础。外伤、炎症等刺激使耳廓软骨发生炎性反应，产生无菌性渗出液。囊肿早期，其外壁系软骨膜，随着病程进展，囊肿外壁软骨膜在渗出液的刺激下增生、变厚、生成新生软骨。新生软骨由薄逐渐增厚，由小逐渐增大，整片软骨由此形成，渗出液不再产生，软骨间的积液形成。随着病程进展，软骨间积液逐渐被吸收，最终机化。新生软骨与囊肿后壁正常耳廓软骨粘连，耳廓变形增厚。同时在光镜下观察囊肿外壁显示：少数血管扩张，部分淋巴细胞浸润，囊壁纤维结缔组织增生。增生的纤维组织中可见薄厚不均匀软骨细胞层，软骨细胞层内可见较小的软骨陷窝及软骨细胞，软骨陷窝和软骨基质呈嗜酸性染色。因此认为耳廓假性囊肿外壁是新生软骨。 研究目的： 1. 通过光学显微镜对耳廓假性囊肿外壁HE染色标本及透射电镜对耳廓假性囊肿外壁软骨进行观察，探讨耳廓假性囊肿外壁软骨来源，明确其为新生软骨，还是退化、变性软骨，亦或坏死软骨。 2. 根据耳廓假性囊肿的不同病理变化进行分期。 研究方法： 本研究获得西安交通大学第二附属医院伦理委员会的批准，在患者知情同意情况下，收集48例（51耳）耳廓假性囊肿外壁作为耳廓假性囊肿组，45例（45耳）慢性化脓性中耳炎住院患者（正常耳廓，无急、慢性感染，无肿瘤等；患者均为慢性化脓性中耳炎术中需取正常耳廓软骨修补鼓膜穿孔）作为正常对照组。手术切除的耳廓假性囊肿外壁部分留作HE染色以及透射电镜标本的制作。慢性化脓性中耳炎患者全麻下行乳突根治术和鼓室成形术过程中取正常耳廓软骨用来修补鼓膜，留取修剪残余的耳廓软骨备用作为HE染色及透射电镜标本制作。光学显微镜下HE染色及透射电镜下标本进行观察并留取图片记录并描述进行结果分析；数据分析使用软件SPSS18.0软件； 年龄以均数±标准差（）表示，用两独立样本t检验比较耳廓假性囊肿组和正常对照组年龄的差异，卡方检验检验两组样本性别差异。 实验结果： 1. t检验比较耳廓假性囊肿组及正常对照组年龄无显著性差异（P＞0.05）；卡方检验比较耳廓假性囊肿组与正常对照组性别无显著性差异（P＞0.05）；耳廓假性囊肿好发于21-60岁人群，其中41-60岁人群更易患该病（52.94%）； 2. 通过病程不同，将耳廓假性囊肿组分为t≤10d、10d＜t≤1m、1m＜t≤2m、2m＜t≤1y、1y＜t≤2y、2y＜t组，对不同病程的囊肿外壁进行显微及超微结构观察发现：由于各种原因引起的软骨间积液即囊液将软骨膜及软骨组织分离，软骨膜受到刺激，其内的骨祖细胞由静止期进入活动期，随着病程的延长，软骨膜逐渐增厚，软骨膜细胞层骨祖细胞分化为成软骨细胞，并进一步转化称为软骨细胞，软骨细胞由幼稚变成熟，软骨层逐渐由薄变厚，至软骨成熟稳定后维持一段时间；故囊肿外壁软骨呈附加性生长过程，囊肿外壁为新生软骨；在病变受到刺激等进一步进展时，软骨组织中软骨细胞核碎裂、核溶解、细胞逐渐坏死，此时软骨呈逐渐坏死状态。 研究结论： 1. 耳廓假性囊肿囊肿外壁软骨是软骨膜附加性生长所形成，为新生软骨；随着病程的延长及进展，囊肿外壁软骨膜下新生软骨由薄变厚，最终软骨细胞逐渐坏死，细胞功能消失，囊肿外壁软骨坏死。 2. 耳廓假性囊肿可根据病理变化进行分期，我们主张分为早、中、晚期。早期即囊壁软骨形成期：病程小于2月；中期即囊壁软骨成熟期：病程介于2月至1年；晚期即囊壁软骨坏死期：病程超过1年。
Background: Auricular pseudocyst is a common disease of otorhinolaryngology. Its etiology and pathogenesis has not been fully elucidated, the disease in the Asian population was significantly more than the disease in the Europe and the United States. Most scholars believe that the incidence of the auricular pseudocyst is associated with the following factors: trauma and local mechanical stimulation; embryos and genetic theory; lysosomal theory; immune factors; perichondrium theory; proteoglycan theory. As the etiology of disease is unclear, treatment methods are diverse, but easy to relapse, which often lead to auricle thickening, deformation. Histopathological features of auricular pseudocyst are still controversial, some scholars believe that the formation of auricular pseudocyst is due to the release of LDH which may lead to the liquefaction of the auricle cartilage. Some scholars think that the cyst wall of the auricular pseudocyst is caused by the cartilage degeneration , degradation or necrosis. Some scholars found that the perichondrium is the foundation of cystic outer wall of the auricular pseudocyst’s formation through the pathological observation. Trauma, inflammation and other stimulation cause the inflammatory reaction of auricular cartilage, resulting in aseptic exudate. Early cysts, the outer wall is the perichondrium. With the progression of the disease, the outer wall of the cyst cartilage is hyperplasia , thickening generating new cartilage by the stimulation of exudate. Newborn cartilage from small to large, from thin to thick, and formare the entire cartilage ultimately, when the exudates terminate at this time to .Finally, interstitial fluid is absorbed, machineized. Hyperplasia of cartilage tissue and auricular cartilage will undergo organization and adhesion, leading to auricle thickening and deformation. At the same time, it is found that fibrous connective tissue is proliferation, and there is a small number of vascular dilatation and lymphocyte infiltration. Hyperplastic fibrous tissue is extremely thin to different thickness, the cartilage lacuna and chondrocytes are small, which is eosinophilic through the observation of the optical microscope. So the outer wall of the auricular pseudocyst is a newborn cartilage rather than cartilage degeneration and degeneration. Objective: 1. The peripheral cartilage of the auricular pseudocyst was observed by optical microscope and the transmission electron microscopy to explore the origin of the cartilage of the anteroposterior pseudocyst, and to identify it as newborn cartilage, degenerate cartilage,or necrotic cartilage. 2. Staging for the auricular pseudocyst according to the different pathological changes. Methods: This study was approved by the Ethical Committee of the Second Affiliated Hospital of Xi'an Jiaotong University. 48 cases (51 ears) of the auricular pseudocyst were collected as the auricular pseudocyst group and 45 patients (45 ears) chronic suppurative otitis media hospitalized patients (normal auricle, no acute, chronic infection, no tumor, etc.; patients are chronic suppurative otitis media surgery to take normal auricular cartilage repair tympanic membrane perforation) as a normal control group. Surgical resection of the auricle pseudocyst of the outer wall was left for HE staining and transmission of electron microscopy specimens. Chronic suppurative otitis media in patients with general anesthesia mastoid radical mastoidectomy and tympanoplasty to take the process of normal auricular cartilage used to repair the tympanic membrane, leaving the pruning residual auricle cartilage as a HE staining and transmission electron microscopy specimens. Optical microscopy under HE staining and transmission electron microscopy specimens were observed and recorded to record and describe the results to analysis; The data analysis using software SPSS18.0; The age was expressed as mean ± standard deviation (). Two different samples were used to compare the age difference between the auricular pseudocyst and the normal control group. The chi-square test was used to test the gender differences between the two groups. Results: 1. There was no significant difference in the age of the auricular pseudocyst and the normal control group by t test (P> 0.05). There was no significant difference in the sex of the auricular pseudocyst and the normal control group by chi-square test (P> 0.05). Auricular pseudocyst occurs in 40-60 years old. 2. Accorrding to the different course of the auricle pseudocyst group is divided into t ≤ 10d, 10d <t≤1m, 1m <t≤2m, 2m <t≤1y,, 1y <t≤2y and 2y <t. The observation of microstructure and ultrastructure of outer walls of the auricle pseudocyst is showed: due to various causes of cartilage fluid that separates perichondrium and cartilage tissue, when the perichondrium is stimulated, the bone progenitor cells from the quiescence phase come to the activities period. With the extension of the course, the periosteum is thickening gradually, cartilage the bone progenitor cells idifferentiate into chondroblasts, which idifferentiate into chondrocytes last, from native to mature, from thin to thick. It will stable a period of time when the cartilage mature . There is karyoeehexis and karyolysis of chondrocytes with the further progress of the disease , cartilage is necrosis at this time. Conclusions: 1. The outer cartilage of auricle pseudocyst is the formed by additional growth of perichondrium ,which is newborn cartilage; with the extension and progress of the course ,the chondrocytes gradually go to necrosis. 2. The pathological staging is divided into early, middle and late stage according to the different pathological changes of the auricular pseudocyst .Early is cartilage formation: less than 2 months; mid-term is cartilage maturity: February to 1 year; late is cartilage necrosis: more than 1 year.