| INTRUDUCTION: About 10%～15% of couples are subfertile, and 40% of these a male factor is involved. The incidence of azoospermia in infertile men is between 5% and 20%. Azoospermia may occur because of reproductive tract obstruction (obstructive azoospermia) or inadequate production of spermatozoa, such that spermatozoa do not appear in the ejaculate(non-obstructive azoospermia). Intracytop lasmic sperm injection(ICSI) and testicular sperm extraction(TESE) have made it possible to treat most forms of azoospermia, and normal embryo development can proceed.OBJECTIVE: This report reviews the basis for the current classification of azoospermia. It argued that many patients could obtain one or many etiological factors. when one patient has many etiological factors, he should be classification by one of accurat factors. Though the use of this classification may be useful in selecting patients for appropriate treatment. We compared the influence of different staining methods of spermatogenic cell morphology analysis, conclud that NF-PICS is an batter technique of spermatogenic cell morphology analysis. We studied the relationship between many factors and spermatogenesis, in order to illuminate the factor of azoospermia. We also study the correlation of semen conventional parameters with accessory sexual gland function and endocrine hormone in order to initially explored the important value of indicator of diagnosis in evaluating azoospermia.PATIENTS: 1665 semen samples were collected, involving control group,oligozoospermic group,severe oligozoospermic and azoospermic group.METHODS:①Azoospermic patients were classified according to etiological factors and WHO male fertile factor diagnosis classification stantard. Then, diffent classification methords were compared.②Serum hormone and genetics analysis were detected.③Semen parameters were analysed. Seminal activity of neutral alpha-glucosidase and fructose concentration were measured using spectrophotometry.④Semen smears were stained according to Modified Papanicolaou, Diff-quick and NF-PICS method. Then, the influence of these three staining methods on the results of spermatogenic cell morphology analysis was compared.RESULTS:①Analyze etiological factors of 223 azoospermia, obstructive azoospermia was 33.6%, non- obstructive azoospermia was 66.4%. According WHO male fertile factor diagnosis classification stantard, iatrogenic disease was 0.4%, systemic disease was 0.9%, congenital anomaly was 45.7%, acquired injury of testis was 1.3%, varicocele was 13.0%, infectious diseases was 12.6%, endocrine disease was 6.3%, obstructive azoospermia was 11.7%.②Comparing with normal group, serum FSH,LH significantly increased in KS patients(P<0.05), but serum T significantly decreased (P<0.05).③Comparing with control group, serum FSH,LH and PRLsignificantly increased in non- obstructive azoospermia group(P<0.01, P<0.05), however, E2 was significantly lower in non- obstructive azoospermia group(P<0.01), No difference was observed between control and obstructive azoospermia groups for serum hormone (P>0.05). Significantly negative correlation was found between sperm density and serum LH,FSH (r=-0.347, P<0.001. r=-0.562, P<0.001), No relationship of the lever of serum PRL,E2,T was found with the sperm density(P> 0.05). Comparing with control group, serum FSH,T significantly increased in testicular volum﹤15ml group(P<0.05). Difference between nomal and oligozoospermia,azoospermia groups were most significant for serum FSH,LH (P<0.05,P<0.01).Comparing with oligozoospermia group, serum FSH,T significantly increased in azoospermia group(P<0.01).④Comparing with control group,seminal plasma zinc significantly decreased in oligzoospermia group(P < 0.05), however, plasma zinc significantly decreased in obstructive azoospermia(P<0.01). No difference was observed between nomal and non- obstructive azoospermia groups(P>0.05). Seminal plasma zinc was positively correlated with the serumT(r=0.225, P<0.05). Significantly negative correlation was found between Seminal plasma zinc and serum LH(r=-0.217, P<0.05), however, no important relationship was obtained between seminal plasma zinc and serum FSH(P>0.05).⑤Difference between nomal and azoospermia groups were most significant for semen volume, seminal liquefaction (P<0.05). No difference was observed in both groups for pH, WBC (P>0.05). Comparing with control group,fructose concentration and activity of neutral alpha-glucosidase were significantly lower in obstructive azoospermia group (P<0.01), but no difference was observed between control and non- obstructive azoospermia groups for fructose concentration and activity of neutral alpha-glucosidase (P>0.05); fructose concentration was positively correlated with the semen volume (P<0.01), however, no important relationship was obtained between fructose concentration and pH. Comparing with the group of fructose positive, seminal volume, pH and neutral alpha– glucosidase were significantly lower in the group of fructose negative(P<0.05). Comparing with the group of fructose negative, FSH,LH were significantly higher(P<0.01), however, T was significantly lower in the group of fructose positive(P<0.05).⑥The lower of Johnson score of testicular biopsy, the more irregular of sertoli cell morphology. Statistical significance of difference of sertoli index was detected between normal spermatogenesis group and the two other groups (hypospermatogenesis and arrest spermatogenesis).CONCLUSION:①When one patient has many etiological factors,he should be classification by one of accurat factors. Though the use of this classification may be useful in selecting patients for appropriate treatment.②Azoospermia have hign prevalence of chromosomal abnormality, for azoospermia, analyzing karyotypes of chromosome by cytogenetic is important excapt.③Y chromosome microdeletio is a vital cause leading to sperm atogenie failure in male infertility.The gene of AZF play important roles in male infertility diagnose.④Testicular volume combined with serum hormone could judge tsticular damage degree.⑤Analying of semen volume,seminal liquefaction,pH and WBC is very important for azoospermia diagnosis and evaluating prognosis.⑥Seminal plasma znic has important effect in testicular spermium.⑦Group of fructose negative could identificate patients of fructose concentration is lower. Fructose qualitation combind with semen volume,pH and neutral alpha-glucosidase could offer evidence for diagnose azoospermia.⑧Determination of seminal biochemistry component is a convenient method for understand function of accessory sex gland. It can help azoospermia diagnosis and spermatic duct occlusion location.⑨Ripe sertoli cell has permanent quantity. Percentage of spermatogenic cell and SEI make the spermatogenesis quantization.This reflect the testicular function.⑩We can consider the three staining methods, Modified Papanicolaou stain, Diff-Quick stain, and NF-PICS stain, as reliable and applicable staining methods to detect the spermatogenic cell morphology. Among the three staining methods, NF-PICS stain is better to be used in examing the spermatogenic cell morphology.
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