精液细胞DNA倍体分析结合生精细胞检查鉴别梗阻性与非梗阻性无精子症

目的：拟通过比较梗阻（OA）和非梗阻性无精子（NOA）症患者精液细胞DNA倍体分析与细胞学检查结果的特点，以期建立一种无创的诊疗方案以鉴别OA与NOA。方法：NOA患者20例和OA患者10例，按照WHO方法进行精液分析诊断，获取其精液标本，液化后离心取沉淀物，1份采用体积分数70％冰乙醇重悬精液沉淀，4~C固定后用PI染色，采用流式细胞仪进行DNA倍体分析。同时，取另1份精液沉淀涂片，进行Diff-Quick染色和免疫细胞化学染色。结果：DNA倍体分析结果表明，20例NOA患者精液中均存在单倍体（1Y）、二倍体（2N）和四倍体（4N）细胞，其中2N细胞含量最高，约占71．25±8．73％。1N细胞的百分比含量为5．46±2．93％，4N细胞含量为3．28±2．54％。10例OA患者精液中有8例只存在2N细胞，含量为71．67±13．09％，未检测到1N和4N，2例未检测到各种倍体细胞。精液细胞学检查结果表明，20例NOA患者中有15例患者精液中可检测出生精细胞，5例患者精液中未检出生精细胞。10例OA患者精液中均未检出生精细胞，其中2例没有任何细胞。结论：精液细胞DNA倍体分析与细胞学检查作为2种无创的检查方法，均可作为评估患者生精功能的辅助诊疗手段，两者结合可作为鉴别OA和NOA的辅助诊断指标。

Identification of Obstructive and Non-obstructive Azoospermia with Seminal Cell DNA Content Analysis and Seminal Spermatogenic Cells Examination

Objective： To establish a non-invasive approach to identify obstructive azoospermia （OA） and non-obstructive azoospermia （NOA） by comparing the results of seminal cell DNA content analysis and seminal spermatogenic cells examination. Methods： Semen samples from 20 NOA and 10 OA patients were examined according to the WHO 5 Guidelines. After liquefaction, the samples were centrifuged to collect spermatogenic cells. For DNA content analysis, the pellet was resuspended with 70% （v/v） ice ethanol and incubated with propidium iodide （PI）. Meanwhile, the pellet smears were performed Diff-Quick staining and immunocytoehemistry. Results： The results of DNA ploidy analysis revealed the presence of haploid （1N）, diploid （2N） and tetraploid （4N） cells, of which the highest percentage of 2N cells, accounting for 71.25 ＋_ 8.73%. The percentage of 1N cells and 4N cells were 5.46 ＋ 2.93% and 3.28 _＋ 2.54%, respectively. Among 10 samples of OA patients, only 8 samples had 2N cells （71.67 ＋ 13.09%） and no cells were detected in 2 patients. Semen cytology results showed that spermatogenic cells could be detected in 15 cases of 20 NOA patients and no spermatogenic cells could be observed in 5 samples of NOA patients. No spermatogenic cells could be detected in samples of OA patients. Conclusion： The results of seminal cell DNA content analysis and seminal spermatogenic cells examination showed that 1N, 2N and 4N cells could be detected in semen of NOA patients and spermatogenie cells could be observed in most semen of NOA patients; meanwhile, only 2N cells or no cells could be detected in semen of OA patients and no spermatogenic cells could be observed in semen of OA patients. Seminal cell DNA content analysis and cytology could be applied for evaluating the status of spermatogenesis as two kinds of non-invasive examination methods. The combination of above methods could be used as diagnostic approach for identification of OA and NOA.