【作者】 高健；

【导师】 段晓刚；

【作者基本信息】 东北师范大学 ， 细胞生物学， 2010， 硕士

【摘要】 精子发生是一个复杂的过程,受多种因素调节,其发生过程大致分睾丸中生成,附睾中成熟,经由输精管、射精管排出体外。精子发生是受各种因素调节,细胞逐步分化的过程,任何一步异常都会导致无精子症。约有10%-15%已婚夫妇不育,其中男性生育功能障碍约占总不育原因的50%。无精子症是男性不育中最严重的一种,发病率约为5%-20%。无精子症有多种原因,诊断有多种方法。用精液生精细胞学检查方法来鉴别诊断梗阻性与非梗阻性（原发性）无精子症的研究备受关注,但尚未引起足够的重视。本文研究无精子症在男性不育患者中的发生率,进一步从不同方面探讨精液生精细胞在诊断梗阻性和非梗阻性无精子症中的意义。本文通过对来自吉林省生殖医学研究所临床基地的2246例不育患者进行研究,检出无精子症患者242例,通过瑞-姬染色分析精液生精细胞,睾丸细针穿刺吸液细胞学分析,测定睾丸体积、精液pH值、精浆果糖、中性α-糖苷酶活性,放射免疫分析法测定血清FSH、LH、PRL、E₂、T水平,常规外周血淋巴细胞染色体标本制备,核型分析,AZF基因检测等,结果如下:无精子症在男性不育患者中发生率为10.77%。在242例无精子症患者精液细胞学分析中,发现有生精细胞存在的患者60例,占无精子症患者24.79%;无生精细胞患者182例,占总无精子症患者的75.21%。在未查到生精细胞的182例患者中,精液细胞学联合多种方法诊断85例为梗阻性无精子症。进一步睾丸细针穿刺细胞学分析发现,83例患者有精子,确诊为梗阻性无精子症。精液细胞学联合多种方法诊断与睾丸细针穿刺细胞学分析的符合率为97.65%（83/85）。与睾丸细针穿刺比较,精液细胞学分析查到生精细胞者60例中,60例均可见曲精小管有未成熟的各级生精细胞或支持细胞,但未发现精子,确诊为非梗阻性无精子症,精液生精细胞检查与睾丸穿刺细胞学分析的符合率为100%（60/60）。精液细胞学分析与睾丸细针穿刺细胞学分析结果经卡方检验,对同一患者精液细胞学检查与睾丸细针穿刺细胞学一致性分析中,生精细胞发育程度一致（P<0.01）。有生精细胞组睾丸体积显著低于对照组（P<0.01）;无生精细胞组睾丸体积、精液量、精液pH值均显著低于对照组与有生精细胞组（P<0.05）。无生精细胞组精浆果糖含量与中性α-糖苷酶活性均显著低于对照组与有生精细胞组（P<0.05）。有生精细胞组FSH、LH显著高于对照组（P<0.01,P<0.05）;而有生精细胞组T显著低于对照组（P<0.05）;无生精细胞组血清FSH、LH均显著高于对照组,两组比较有显著性差异（P<0.05）,而无生精细胞组T显著低于对照组（P<0.05）;无生精细胞组FSH显著低于有生精细胞组（P<0.05）。在无精子症中,克氏症患者检出率为19.01%（46/242）。在46例克氏症患者中,检出精原细胞33例（71.74%）,初级精母细胞20例（43.48%）,次级精母细胞11例（23.91%）,精子细胞3例（6.52%）。AZF缺失检出率占7.85%（19/242）。在AZFb缺失组,精液细胞学分析发现初级精母细胞。在AZFc缺失组,精液细胞学分析有部分患者可检测出各级生精细胞,而有部分未见各级生精细胞。在AZFd缺失组,未见各级生精细胞。以上结果表明:精液生精细胞在无精子症病因诊断中,是非常有价值的非创伤性指标,对区分梗阻性无精子症和非梗阻性无精子症,及梗阻性无精子症的定位诊断有很高的预示性价值,可以替代睾丸活检,甚至可替代睾丸细针穿刺。精液生精细胞与睾丸体积、精液量、精液pH值、精浆果糖含量、中性α-糖苷酶活性、血清FSH、LH、T在诊断无精子症上具有一致性,且相互补充,可联合应用诊断无精子症类型。克氏症患者精液中有不同程度的生精细胞,精液生精细胞检查能一定程度上反应克氏症患者的精子生成能力,可作为临床评估生精功能的方法之一。AZFb、AZFc、AZFd缺失患者精液中有不同程度的生精细胞,精液生精细胞检查能一定程度上反应AZF缺失患者的精子生成能力,可作为临床评估生精功能的方法之一。更多还原

【Abstract】 Spermatogenesis is a complicated process, and is regulated many factors. Any abnormal factor can lead to azoospermatism. About 10%-15% of married couple can be infertile, 50% of which is male infertility. Azoospermatism is the most important one in male infertility; its incidence rate is about 5%-20%. The diagnose of azoospermatism have many methods. We have paid close attention to detecting spermatogenic cell of ejaculum in diagnosing obstructive or non-obstructive azoospermatism, but don’t think highly of this method. In the current study, we investigated incidence rate of azoospermatism in male infertility, and explored the significance of detecting spermatogenic cell of ejaculum in diagnosing azoospermatism.In the current study, several comparative studies were carried out to clarify whether detecting spermatogenic cell of ejaculum in diagnosing obstructive or non-obstructive azoospermatism or not. 2246 patients were studied. 242 cases was azoospermatism. To analyze semen spermatogenic cell, Wright-Giemsa staining method was used. Orchid-fine needle aspiration was used. We detected testis volume, pH of semen, seminal plasma fructose, and neutral a-glycosidase. To detected level of serum FSH、LH、PRL、E₂、T, radio immunoassay was used. Karyotype analysis of chromosome was used. Deletion of AZF gene was detected. Results are as follows:The incidence rate of azoospermatism in male infertility was 10.77%. In 242 cases of azoospermatism, 60 cases （24.79%） had spermatogenic cell in semen, 182 cases （75.21%） didn’t have spermatogenic cell in semen. In 182 cases of non- spermatogenic cell in semen, Obstructive azoospermatism of 85 cases is diagnosed by spermatogenic cell with many methods. By orchido-fine needle aspiration methods, 83 cases have sperm, and are obstructive azoospermatism. The diagnostic correspondent rate of two methods was 97.65%. In 60 cases of spermatogenic cell in semen, the seminal spermatogenic cells examination was coincident with the result of orchido-fine needle aspiration methods at the spermatogenic cells developmental level in the same patient （P<0.01）. By orchido-fine needle aspiration methods, 60 cases have not sperm, and are non-obstructive azoospermatism. The diagnostic correspondent rate of two methods was 97.65%.Orchido-volume of the group with spermatogenic cell was significantly lower than that of fertile group （control group）（P<0.01）. Orchido-volume, seminal fluid volume, seminal fluid pH of the group without spermatogenic cell were all significantly lower than that of fertile group and the group with spermatogenic cell （P<0.05）. The content of seminal plasma fructose and neutralα-glycosidase activity of the group without spermatogenic cell were both significantly lower than that of fertile group and the group with spermatogenic cell （P<0.05）. Serum FSH, LH of the group with spermatogenic cell were significantly higher than that of fertile group （P<0.01, P<0.05）, but serum T of the group with spermatogenic cell was significantly lower than that of fertile group （P<0.05）. Serum FSH, LH of the group without spermatogenic cell were significantly higher than that of fertile group （P<0.05）, but serum T of the group without spermatogenic cell was significantly lower than that of fertile group （P<0.05）. Serum FSH of the group without spermatogenic cell was significantly lower than that of the group with spermatogenic cell （P<0.05）.In azoospermatism, detection rate of Klinefelter symptom is 19.01%. In 46 cases of Klinefelter symptom, there are 33 cases spermiogonium, 20 cases primary spermatocyte, 11 cases secondary spermatocyte, 3 cases spermatid in semen. The detection rate of AZF deletion is 7.85%. The group of AZFb deletion, there is primary spermatocyte in semen. The group of AZFc deletion, part of the group has all level spermatogenic cells; the other has not all level spermatogenic cells. The group of AZFd deletion, there is not all level spermatogenic cells.All result indicates that the seminal spermatogenic cell examination is an extremely untraumatic, ideal method which can both reflect the spermatogenic status in the testis and prompting obstructive situation of seminal ducts, and can take the place of. orchido-biopsy, orchido-fine needle aspiration. The seminal spermatogenic cell examination was coincident with orchido-volume, orchido-volume, seminal fluid pH, seminal plasma fructose, neutral a-glycosidase, serum FSH, LH, T, and can be complementation each other. Patients of Klinefelter symptom and AZFb, AZFc, AZFd deletion have different spermatogenic cell in semen, and the seminal spermatogenic cell examination can reflect capability of gonepoiesis, can be one of clinical assessment methods of these patients’gonepoiesis.更多还原