| ForewordInfertile couples which are caused by male factor is about 25～40%. So thepeople pay more and more attention to the semen quality. Semen parameters areimportant index which could reflect the semen quality, while the morphology is aimportant part of semen parameters. Sperm morphology plays a great role inevaluating the sperm fertilizing ability and predicting the outcomes of assistedreproductive technology (ART). In the World Health Organization (WHO) guideline:when the proportion of abnormal morphology sperm is above the 70% in semen, it iscalled teratozoospermia. The poorer the sperm morphology is, the lower the naturalpregnancy is achieved. Some teratozoospermia patients have to resort to ART tosatisfy their need of fertility. So we must evaluate precisely the sperm morphology,but also find the suitable sperm preparation method and assisted reproductivetechnology which could fit the teratozoospermia person, in order to improve theirfertility.ObjectiveTo find the simple, quick and precise sperm morphology stain method which issuitable for clinical analysis to facilitate the assessment of sperm morphology; Toseek the appropriate sperm preparation method and assisted reproductive technologyfor teratozoospermia person in order to improve the clinical pregnancy rate of suchpatients.1. Comparing Wright-Giemsa stain with the WHO recommended Papanicolaou stain on sperm morphology evaluation, hoping to find a relatively simple and effectivesperm morphology stain method which can replace Papanicolaou stain.2. Comparing the effect of direct swim-up and percoll density gradient centrifugationcombined swim-up methods on the treatment of teratozoospermia, trying to find asuitable sperm preparation method for such patients.3. Assessing the influence of sperm morphology on the fertilization rate, cleavagerate and high-quality embryos rate of in vitro fertilization, and finding theappropriate method of ART for teratozoospermia couples.Methods1. Semen smear were stained by Wright-Giemsa and Papanicolaou staining andanalysised the result of the two methods.2. The changes of routine semen parameters, teratosperm proportion, sperm nucleusDNA maturity, acrosome integrity, mitochondrial membrane potential, the level ofreactive oxygen species(ROS) before and after the two methods prepared wereanalysised by computer-assisted semen analysis(CASA), sperm morphology stain,acridine orange stain and flow cytometry (FCM).3. Analysised the effect of sperm morphology on the rate of fertilization, cleavageand high-quality embryos in conventional in vitro fertilization(IVF) andintracytoplasmic sperm injection(ICSI).Results1. These two stain methods had no difference in assessmenting the spermmorphology (P＞0.05. In evaluating the normal morphology and teratospermia,they had a positive correlation, the correlation coefficient were 0.984, 0.968respectively(P＜0.01). 2. After preparation, the sperm motility increased and teratosperm proportiondecreased significantly, the results of percoll density gradient centrifugationcombined swim-up were better than the those of direct swim-up(34.8±11.2% vs57.4±13.7%, 61.4±12.8; 53.7±5.8% vs 81.7±6.2%, 85.7±5.7%; 78.8±8.2% vs70.2±12.0%, 64.3±11.5% respectively, P＜0.01). Direct swim-up had higherproportion of double strands-DNA sperm (72.0±20.3% vs 68.9±20.6%, P＜0.01),while the percoll density gradient centrifugation combined swim-up containedlower ROS(146.7±65.2 vs 142.0±61.8, P＜0.05). There was no difference aboutacrosome integrity and mitochondrial membrane potential.3. In IVF treatment, the fertilization rate in normal morphology group was higherthan in abnormal morphology group(0.72±0.21 vs 0.55±0.29), the difference wassignificant(P＜0.05); with regard to the cleavage rate and high-quality embryosrate, the normal morphology group was also higher than the abnormal morphologygroup, but there were no significant differences(P＞0.05). In ICSI treatment, therewere no significant differences about the the rate of fertilization, cleavage andhigh-quality embryos between the normal and abnormal sperm morphologygroups(P＞0.05). Besides, in the normal sperm morphology group, the IVF andICSI got the same rate of fertilization, cleavage and high-quality embryos(P＞0.05).In teratozoospermia group, the fertilization rate in ICSI treatment was higher thanIVF treatment(0.72±0.20 vs 0.55±0.29), the difference was significant(P＜0.05);the rate of cleavage and high-quality embryos was higher in ICSI treatment, butthere were no significant differences(P＞0.05).Conclusions1. Wright-Giemsa stain had no difference in assessmenting the sperm morphologycompared to Papanicolaou stain, but the Wright-Giemsa stain was moreconvenient, time-saving, fast, economic. So it could replace Papanicolaou stain toassess the sperm morphology. 2. These two methods could improve the sperm function effectively. As to theteratozoospermia, we might choose the proper separation method according to themethod of ART.3. For normal sperm morphology, we could choose the suitable ATR methodaccording to the cause of their infertility; while for teratozoosperm couples, ICSItreatment was better which could increase its fertility rate and improve theirreproductive outcome.
|
PDF Full Text Request