Can inhibin-B predict the outcome of microsurgical epididymal sperm aspiration in patients with suspected primary obstructive azoospermia？

Aim： To evaluate whether inhibin-B can predict the outcome of a microsurgical epidymal sperm aspiration （MESA） procedure in patients with suspected primary obstructive azoospermia （OA） and if inhibin-B can replace testicular biopsy in the diagnostic work-up of these patients. Methods： Inhibin-B levels and testicular biopsy scores were related to the outcome of MESA in 43 patients with suspected primary OA. MESA was considered to be successful when epididymal sperm could be identified during the procedure. Results： Spermatozoa were present in the epididymal aspirate in 28 out of the 43 patients （65%）. lnhibin-B values were not significantly different in patients with successful or unsuccessful MESA. The modified Johnsen score, however, was significantly lower in patients with unsuccessful MESA （P = 0.003）. A rete testis obstruction or epididymal malfunctioning was found in 15% of patients with suspected primary OA, reflected by unsuccessful MESA despite normal inhibin-B levels and normal testicular histology. Conclusion： Inhibin-B cannot replace testicular biopsy as a diagnostic tool in the work-up of patients with suspected primary OA. Testicular biopsy is useful in identifying patients with spermatogenic arrest, who might have normal inhibin-B values.

Successful pregnancy and birth after intrauterine insemination using caput epididymal sperm by percutaneous aspiration

<abstract>Aim: To manage male infertility with obstructive azoospermia by means of percutaneous epididymal sperm aspiration (PESA) and intrauterine insemination (IUI). Methods: Ninety azoospermic patients with congenital bilateral absence of the vas deferens (BAVD, n=58) or bilateral caudal epididymal obstruction (BCEO, n=32) requesting for fine needle aspiration (FNA), PESA and IUI were recruited. The obstruction was diagnosed by vasography and determination of the fructose, carnitine and alpha-glucosidase levels in the seminal fluid. Results: The mean sperm motility, density, abnormal sperm and total sperm count of the caput epdidymis were 16 %±22 %, (12±31) ×106/mL, 55 %±36 % and (16±14)×106, respectively. In the 90 couples, a total of 74 PESA procedures and 66 cycles of IUI were performed. Three pregnancies resulted, including one twin pregnancy giving birth to two healthy boys, one single pregnancy with a healthy girl and another single pregnancy aborted at week 6 of conception. The pregnancy rate per IUI cycle was 4.5 %. Conclusion: The birth of normal, healthy infants by IUI using PESA indicates that the caput epididymal sperm possess fertilization capacity. The PESA-IUI programme is a practical and economical procedure for the management of patients with obstructive azoospermia.

Microsurgical epididymal sperm aspiration： indications, techniques and outcomes

Microsurgical epididymal sperm aspiration （MESA） refers to retrieval of sperm-containing fluid from optimal areas of the epididymis that are selected and sampled using high-power optical magnification provided by an operating microscope. Retrieved sperm are subsequently used for intracytoplasmic sperm injection （ICSI） to induce fertilization and pregnancy. MESA is considered by many experts to be the gold standard technique for sperm retrieval in men with obstructive azoospermia given its high yield of quality sperm, excellent reported fertilization and pregnancy rates, and low risk of complications. However, MESA must be performed in an operating room, requires microsurgical skills and is only useful for reproduction using ICSI. Herein we present an overview of the evaluation of candidate patients for MESA, the technical performance of the procedure and the outcomes that have been reported.

Medium Term Survival of Bovine Caudal Epididymal Sperm under Conditions Based on Epididymal Fluid Characteristics

Fresh semen has a very limited life span after it leaves the gonad. The relative research shows that the epididymal plasma provides a special environment to spermatozoa for survival. In this study, the osmolarity and protein concentration of caput and caudal bull epididymis were analyzed. Bovine caudal epididymal sperm were incubated in different protein and sorbitol combinations of CEP-2 diluents at 4 ℃ for 120 h and sperm viability was assessed. Under this condition, sperm viability was greater than 60% after 5 d. The osmolarity of caput and caudal epididymis were （287.0 ± 13.7） and （310.8 ± 17.0 ） mOsm, respectively. The protein concentration of caput and caudal epididymis were （ 37.43 ± 12.55 ） and （50.58±11.08） mg/mg, respectively. This may have an application in the animal production industry.

Minimal volume vitrification of epididymal spermatozoa results in successful in vitro fertilization and embryo development in mice

This study compared three cryopreservation protocols on sperm functions, IVF outcomes, and embryo development. Epididymal spermatozoa cryopreserved using slow-cooling （18% w/v raffinose, RS-C） were compared with spermatozoa vitrified using 0.25 M sucrose （SV） or 18% w/v raffinose （RV）. The motility, vitality, and DNA damage （TUNEL assay） of fresh control （FC） spermatozoa were compared with post-thawed or warmed RS-C, RV, and SV samples. Mouse oocytes （n = 267） were randomly assigned into three groups for insemination： RV （n = 102）, RS-C （n = 86）, and FC （n = 79）. The number and the proportion of two-cell embryos and blastocysts from each treatment were assessed. Sperm motility （P 〈 0.01） and vitality （P 〈 0.05） were significantly reduced after vitrification compared with slow-cooled spermatozoa. However, DNA fragmentation was significantly reduced in spermatozoa vitrified using sucrose （15 - 1.8% [SV] vs 26 - 2.8% [RV] and 27 - 1.2% [RS-C]; P 〈 0.01）. Although the number of two-cell embryos produced by RS-C, RV, and FC spermatozoa was not significantly different, the number of blastocysts produced from two-cell embryos using RV spermatozoa was significantly higher than FC spermatozoa （P = 0.0053）. This simple, small volume vitrification protocol and standard insemination method allows successful embryo production from small numbers of epididymal spermatozoa and may be applied clinically to circumvent the need for ICSI, which has the disadvantage of bypassing sperm selection.

In-vitro effect of Peganum harmala total alkaloids on spermatozoa quality and oxidative stress of epididymal ram semen

Objective:To determine the in-vitro effect of the total alkaloid extract of Peganum(P.)harmala seeds on ram epididymal sperm.Methods:Semen was divided into six groups according to the following concentrations of the P.harmala total alkaloids:1,5,10,50,and 100μg/mL,and the control group.The samples were incubated at ambient temperature(21℃-24℃)for 24 h,and analyzed in terms of motility,membrane integrity,and oxidative status.Results:The sperm kinematic parameters,i.e.straight-line velocity,curvilinear velocity,average path velocity,were significantly higher when treated with P.harmala at concentrations ranging from 1 to 10μg/mL compared to the control group(P<0.05).In addtion,the highest amplitude of the lateral head displacement value was found in the groups treated with concentrations 1 and 5μg/mL of P.harmala compared to the control group(P<0.05).Total and progressive motilities showed that the extracts at 1,5,and 10μg/mL exhibited a high percentage after 24 h of incubation.The effect of P.harmala extracts on the membrane integrity of ram epididymal sperm was concentration-dependent and significantly different compared to the control group(P<0.05).Non-significantly lower lipid peroxidation levels were observed after 24 h of incubation of ram epididymal sperm treated with concentrations 1,5,and 10μg/mL of P.harmala extracts compared to the control group(P>0.05).Conclusions:Low concentrations(1-10μg/mL)of P.harmala extracts stimulate sperm motility,preserve membrane integrity and protect ram spermatozoa from lipid peroxidation.

Cryopreservation of a Small Number of Human Sperm within Empty Zona Pellucidae

Objective To investigate the empty zona pellucida for use in the cryopreservation of human sperm. Materials & Methods Human and hamster zona pellucidae were evacuated and injected with testicular, epididymal and ejaculated sperm. The zona pellucidae with sperm were cryopreserved. Results After thawing, zona pellucidae were easily found, and sperm inside zona pellucidae were also easily observed. There were no differences in post-thaw motility and vitality between ejaculated and epididymal sperm groups (P>0.05), but these two parameters were lowered in testicular sperm group compared to both ejaculated and epididymal sperm (P<0.01). No significant difference was observed in post-thaw motilities among 6%, 7.5%, 9% glycerol concentrations (P>0.05). In addition, obvious differences in post-thaw motilities were not found between human and hamster empty zona pellucidae (P>0.05). Conclusion An evacuated zona pellucida is an ideal vehicle for the cryopreservation of a small number of human sperm.