| Objective: For approach the method of sperm retrieval during ICSI therapy, the relative of the quantitative of expression of protamine-2 and hspA2 mRNA of multipoint testicular tissue and the results of sperm extractoion in patients with azoospermia were investigated.Clinical Material and Methods: 38 male infertility patients with azoospermia were selected. Two specimens, about 4~5mm~3 in size, were gained from every side testis of the patients. Every specimen were divided three portions, about 3mm~3, 1mm~3 and 0.5~0.8mm~3 in size, were used for general pathologic inspected, for isolated sperm, and for extracted mRNA, respectively. The morphologic change of the testis pathological section were observed and the function of spermatogenesis were evaluated with the later period spermatids counting. The sperm isolated were done with invert microscope at 400 ×. Once found one sperm in the specimen, then isolated sperm successful were thought in this tissue. If all of one specimen were checked, the sperm couldn't be found, then no sperm in this tissue were thought. The mRNA of the protamine-2, hspA2 and β-actin were amplifed with RT-PCR. The production of amplificationwere separated with electrophoresis. The density of amplified production of the protamine-2, hspA2 by electrophoresis were compared with the density of amplifed production, in same condition, of the (3-actin by electrophoresis, respectively. The relative quantitative of expression of the protamine-2 and hspA2 were calculated. The later period spermatids counting, the successful rate of the sperm retrieval and the quantitative of expression of the protamine-2 and hspA2 mRNA of the superior and inferior position of every side testis were contrasted. The relative of the values of the later period of spermatids counting and the successful rate of sperm extraction were analyzed. The relative of the value of the quantitative of expression of the protamine-2 and hspA2 mRNA and the successful rate of sperm extraction were also analyzad.Results: The pathologic results of 38 cases azoospermia showed 12 with hypospermatogenesis, 6 with spermatid arrest, 5 with spermatocyte arrest, 4 with Sertoli cell-only syndrome and 11 cases with obstructive azoospermia. The successful rate of sperm retrieval, the later period of spermatids counting and the quantitative of expression of the protamine-2 and hspA2 mRNA of superior and inferior posisition of every side testis revealed different for individual of nonobstructive azoospermic patients. These difference distributed on superior and inferior posisition of every side testis showed irregularity. Therefor, the difference of the later period of spermatids counting, the successful rate of sperm retrieval, and the quantitative of expression of the protamine-2 and hspA2 mRNA of superior andinferior posisition of every side testis show no statistical significant in nonobstructive azoospermic group (P>0.05). Althoough almost of the every position on the tsetis could find sperm in obstructive azoospermic group, other indexes, the later posterior of sperm counting and the quantitative of expression of the protamine-2 and hspA2 mRNA of obstructive azoospermia group also showed similar tendency to nonobstructive azoospermic group. The successful sperm extraction or not of position corresponding of the quantitive of expression of protamine-2 revealed 1.45±0.22 and 0.59±0.57 (PO.05), but results of the quantitive of expression of hspA2 mRNA showed no difference (P>0.05). The total successful rate sperm retrieval of two position (right: 48% , left: 39%) higher than Mono position (right: 22% and 37% ; left: 26% and 30%) in the azoospermia group (PO.05).Conclusion: It were suggested that Multipoint testicular tissue sperm extraction of nonobstructive azoospermia for ICSI therapeusis may improve the successful rate of the sperm retrieval. Acting as a predict marker for results of testicular tissue sperm extraction, the protamine-2 were worth further study.
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