Testicular spermatozoon is superior to ejaculated spermatozoon for intracytoplasmic sperm injection to achieve pregnancy in infertile males with high sperm DNA damage

The purpose of this study was to compare the clinical outcome of testicular spermatozoon versus ejaculated spermatozoon in the treatment of infertile males with high sperm DNA damage, referred as sperm DNA fragmentation index (DFI), that attending intracytoplasmic sperm injection (ICSI) programme in terms of clinical pregnancy, births delivered as the primary and pregnancy loss and embryo fertilisation as the secondary outcome. A total of 102 males fulfilling the inclusion criteria were enrolled in the present study. Of the 102 males, 61 infertile males underwent testicular spermatozoon combined with ICSI while the remaining 41 males applied ejaculated spermatozoa in their first ICSI cycles, and the data of them were collected and analysed. In a 18‐month follow‐up, testicular spermatozoon achieved higher pregnancy rate and deliver rate than those in ejaculated sperm group (pregnancy rate, 36% vs. 14.6%, p = 0.017; deliver rate, 38.5% vs. 9.8%, p = 0.001). Nevertheless, there were no significant differences in the number of oocytes aspirated and number of embryos transferred between the two groups. Additionally, the fertilisation rate in the testicular sperm study cohort (70.4%) was also similar to that in the ejaculated sperm group (75.0%). Based on the current data, we conclude that testicular spermatozoon is the prior option in the treatment of infertile males with high sperm DFI in ICSI programme. More high‐quality studies with larger samples size are needed in the future due to the relative small size and the nonrandomized design of the present study.     

Comparison of intracytoplasmic sperm injection outcomes between spermatozoa retrieved from testicular biopsy and from ejaculation in cryptozoospermic men

The infrequent presence of spermatozoa in cryptozoospermic men ejaculate is a limiting factor in the treatment of them. Sometimes, this consideration impels us to apply meticulous microscopic search in ejaculate or testicular sperm extraction (TESE) method. The aim of this study was to assess putative effectiveness of sperm origin, ejaculated or testicular, in cryptozoospermia treatment. In this context, were evaluated intracytoplasmic sperm injection (ICSI) outcomes in two parameters including fertilisation rate (2PN) and embryo quality, independently. We compared the outcome in two groups: patients who underwent ejaculate/ICSI and ones who underwent TESE/ICSI process. Nineteen ICSI cycles performed with testicular spermatozoa and the rest of cycles (n = 208) carried out with ejaculated spermatozoa. Result analysis showed similar fertilisation rate between testicular and ejaculated spermatozoa (respectively, 60% versus 68%, P ≥ 0.05). Also, on the other hand, embryo quality did not show significant differences between two groups, except grade A with low significance. With regard to almost equal performance of both methods in results and being invasive of TESE as surgical sperm retrieval method, the use of ejaculated sperm more than testicular sperm should be recommended in patients with cryptozoospermia whenever possible.     

Testicular versus ejaculated spermatozoa in ICSI cycles of normozoospermic men with high sperm DNA fragmentation and previous ART failures

As a part of male assessment, conventional sperm parameters including morphologic features have been dedicated as major factors influencing fertilisation and pregnancy rates in assisted reproductive technology (ART). Genomic integrity of spermatozoa has also been found to influence fertility prognosis, and hence, sperm DNA fragmentation index (DFI) has been adopted by many centres to document this entity. Despite several suggested approaches, there is lack of universal consensus on optimising fertility outcomes in males with high sperm DFI. In this context, the results from cycles using testicular spermatozoa (TESA) obtained by aspiration were compared with those of ejaculated spermatozoa (EJ) in normozoospermic subjects with high sperm DFI and previous ART failures. Clinical (41.9% versus 20%) and ongoing pregnancy rates (38.7% versus 15%) were significantly better and miscarriages were lower in TESA group when compared to EJ group. Sperm DFI should be a part of male partner's evaluation following unsuccessful ART attempts. When high DFI is detected (>30%), ICSI using testicular spermatozoa obtained by TESA seems an effective option particularly for those with repeated ART failures in terms of clinical, ongoing pregnancies and miscarriages even though conventional sperm parameters are within normal range.     

Intracytoplasmic sperm injection outcomes with cryopreserved testicular sperm aspiration samples

Intracytoplasmic sperm injection (ICSI) may be performed with testicular frozen–thawed spermatozoa in patients with nonobstructive azoospermia (NOA). Sperm retrieval can be performed in advance of oocyte aspiration, as it may avoid the possibility of no recovery of spermatozoa on the day of oocyte pickup. There are few studies available in the literature concerning the use of frozen–thawed spermatozoa obtained from testicular sperm aspiration (TESA). To evaluate the effects and the outcomes of ICSI with frozen–thawed spermatozoa obtained by TESA, we performed a retrospective analysis of 43 ICSI cycles using frozen–thawed TESA. We obtained acceptable results with a fertilisation rate of 67.9%, an implantation rate (IR) of 17.1%, and clinical and ongoing pregnancy rates of 41.9% and 37.2% respectively. The results of this study suggest that performing ICSI using cryopreserved frozen–thawed testicular spermatozoa with TESA as a first option is a viable, safe, economic and effective method for patients with NOA.     

Virtual azoospermia and cryptozoospermia--fresh/frozen testicular or ejaculate sperm for better IVF outcome?

Men diagnosed as having azoospermia occasionally have a few mature sperm cells in other ejaculates. Other men may have constant, yet very low quality and quantity of sperm cells in their ejaculates, resulting in poor intracytoplasmic sperm injection (ICSI) outcome. It has not been conclusively established which source of sperm cells is preferable for ICSI when both ejaculate and testicular (fresh or frozen) sperm cells are available. It is also unclear whether there is any advantage of fresh over frozen sperm if testicular sperm is to be used. We used ejaculate, testicular (fresh or frozen) sperm cells, or both for ICSI in 13 couples. Five of these couples initially underwent ICSI by testicular sperm extraction, because the males had total azoospermia, and in later cycles with ejaculate sperm cells. Ejaculate sperm cells were initially used for ICSI in the other 8 patients, and later with testicular sperm cells. The fertilization rate was significantly higher when fresh or frozen-thawed testicular sperm cells were used than when ejaculated sperm cells were used. Likewise, the quality of the embryos from testicular (fresh and frozen) sperm was higher than from ejaculated sperm (65.3% vs 53.2%, respectively, P < .05). The use of fresh testicular sperm yielded better implantation rates than both frozen testicular sperm and ejaculate. Therefore, fresh testicular sperm should be considered first for ICSI in patients with virtual azoospermia or cryptozoospermia because of their superior fertility.     

Influence of motility and vitality in intracytoplasmic sperm injection with ejaculated and testicular sperm

The vitality of spermatozoa used for intracytoplasmic sperm injection (ICSI) is a crucial factor for fertilization, establishment and outcome of a pregnancy in assisted reproductive technique cycles. The sperm origin may also be a limiting factor, although little is known about this issue. It is known that the motility of injected spermatozoa and their origin from ejaculate or testicular biopsies are important predictors in terms of fertilization, pregnancy and birth rates. Oocytes of patients in 2593 cycles were retrieved in our in vitro fertilization programme and inseminated via ICSI. We used motile (group 1, n = 2317) or immotile ejaculated spermatozoa (group 2, n = 79), motile sperm retrieved from testicular biopsies (group 3, n = 62) and immotile spermatozoa from testicular biopsies (group 4, n = 135). Female age and number of oocytes retrieved did not differ significantly among the groups. The fertilization rates were as follows: 67.1% in group 1, 49.8% in group 2, 68.3% in group 3 and 47.8% in group 4. The pregnancy rates in cases where three embryos had been transferred amounted to 35.7% in group 1, 17.3% in group 2, 38.3% in group 3 and 20.5% in group 4. The embryo quality showed no differences between groups 1 and 3 (14.5), and between groups 2 (11.8) and 4 (10.8). The abortion rate was similar in groups 1-3, but increased in group 4 (26.6%, 27.3%, 31.6% and 55.5%). Irrespective of their origin, the fertilization potential of injected spermatozoa was found to be influenced by motility. The resulting pregnancy and birth rates, i.e. the potential of the resulting embryos to implant and to achieve viable pregnancies, seem to be additionally dependent on the sperm origin. This was well shown by declining rates when spermatozoa in a relatively early stage of maturity had been used. We see increasing evidence that the degree of sperm maturity has an important impact on the outcome of ICSI. In obstructive azoospermia, spermatozoa retrieved from the epididymis should be used rather than testicular biopsy spermatozoa, or testicular sperm should be preincubated in culture medium before ICSI.     

Clinical use of pentoxifylline for activation of immotile testicular sperm before ICSI in patients with azoospermia

The testicular sperm from biopsy and frozen/thawed tissue are frequently immotile. The purpose of our retrospective study was to assess the effect of short exposure of testicular samples with only immotile sperm to pentoxifylline (PF)-sperm motility stimulator. In 77 of 294 (26.2%) testicular sperm ablation/testicular sperm extraction-intracytoplasmic sperm injection (TESA/TESE-ICSI) cycles in patients with azoospermia, only immotile sperm were found in biopsies even after 2 hours of incubation of tissue in the medium. These 77 cycles were divided into 2 groups. In group 1 (cycles between 1999 and 2001; n = 30), ICSI was performed with untreated immotile sperm. In group 2 (cycles between 2002 and 2004; n = 47), immotile testicular sperm were treated for 20 minutes with pentoxifylline (PF) (1.76 mM) before ICSI. Both groups had the same proportion of ICSI cycles with fresh, frozen/thawed, and aspirated testicular sperm. The overall pregnancy rate of TESA/TESE-ICSI did not vary during the study period. In 45 of 47 (95.7%) testicular samples with total immotility, the sperm started to move 20 minutes after PF treatment. The mean time required for ICSI was shortened in the PF group (30 minutes [minimum 10, maximum 90] vs 120 minutes [minimum 60, maximum 240]) due to easier identification of motile sperm. In comparison with the nontreated group, the PF group had a higher fertilization rate (66% vs 50.9%; P < .005) and mean number of embryos per cycle (4.7 +/- 3.3 vs 2.7 +/- 2.1; P < .01). The clinical pregnancy rate per cycle in PF and non-PF groups was 38.3% and 26.7%, respectively. By using PF in cases of only immotile testicular sperm we can cause movement of testicular sperm, allow easier identification of vital sperm, shorten the procedure, improve fertilization rates, and increase the number of embryos.     

Spermatozoa retrieved by electroejaculation: Should we prefer fresh or cryopreserved spermatozoa for intracytoplasmic sperm injection?

We aim to evaluate our experience, comparing intracytoplasmic sperm injection (ICSI) outcomes of cycle using fresh versus thawed electroejaculated spermatozoa. All consecutive couples undergoing ICSI cycles using electroejaculated spermatozoa, during a 16-year period, were evaluated. Embryological/laboratory variables of the ICSI cycles were assessed and compared between those utilising fresh (fresh group) versus thawed (thawed group) electroejaculated spermatozoa. Fifty-seven couples were evaluated, 30 used a fresh electroejaculated spermatozoa in 55 ICSI cycles, while 27 used a thawed sperm sample in 41 ICSI cycles. There were no in-between group differences in the mean numbers of oocytes retrieved per oocyte retrieval nor the percentage of MII oocytes. The fresh group demonstrated significantly higher fertilisation (71.5% vs. 64.1%, respectively, p < .05), top-quality embryos (66.5% vs. 54.9%, respectively, p < .02), clinical pregnancy per transfer (41.3% and 21.2%, respectively, p < .05) and cumulative clinical pregnancy (58.2% vs. 26.8%, respectively, p < .001) rates, as compared to the thawed group. Independent of the source of spermatozoa used, no pregnancy was achieved following ICSI utilising immotile spermatozoa. In conclusion, ICSI cycles using ejaculated spermatozoa of patients suffering from neurologic or psychogenic anejaculation are reassuring. The use of fresh ejaculated spermatozoa retrieved on the day of the female spouse oocyte retrieval might improve outcome. Whenever a thawed electroejaculated spermatozoa yield no motile spermatozoa, emergency electroejaculation is mandatory.     

Intracytoplasmic sperm injection by testicular sperm in patients with aspermia or azoospermia after cancer treatment

The aim of this retrospective study was to evaluate the efficiency of testicular biopsy and intracytoplasmic sperm injection (ICSI) in patients with aspermia or non-obstructive azoospermia (NOA) after cancer treatment. From 1996 to 2003, 30 men with a history of cancer, affected by aspermia or NOA and without sperm cryopreserved before cytotoxic treatment underwent testicular sperm extraction (TESE). In these men, clinical, hormonal and histological characteristics were compared; 13 underwent 39 TESE-ICSI cycles using frozen-thawed testicular spermatozoa (TESE-ICSI group). In the same period, 31 ICSI cycles were performed in 20 men with aspermia or NOA using ejaculated sperm frozen before cancer treatment (ejaculated sperm-ICSI group). Fertilization, blastocyst development, pregnancy and miscarriage rates were compared between the groups. Testicular volume, serum follicle-stimulating hormone level and Johnsen score indicated complete although reduced spermatogenesis in men with aspermia and abnormal spermatogenesis in men with NOA. After TESE, sperm retrieval was positive in 92% of men with aspermia and 58% of men with NOA. In TESE-ICSI patients with NOA a significantly lower proportion of embryos developed to the blastocyst stage than in patients with aspermia and in those after ICSI with frozen-thawed ejaculated sperm (23% vs. 43% and 47%, p = 0.03 and p < 0.01 respectively). In all groups the miscarriage rates were high; in patients with aspermia and NOA, characterized by increased age, the miscarriage rate tended to be higher in spite of similar female age and female indications of infertility. In patients affected by aspermia or NOA after cancer treatment and without sperm cryopreserved before treatment, TESE-ICSI using testicular sperm provide a chance to father a child.     

无精子症病人100例取精方法及妊娠结局

目的 :回顾性分析 2 0 0 1年 1月～ 2 0 0 2年 1月在生殖中心行卵胞质内单精子注射 (ICSI)治疗的 10 0例无精子症男性的治疗结果。　方法 :经皮附睾精子抽吸术 (PESA)或睾丸精子抽提术 (TESE)获得精子 ,女方进行常规超排卵。分析激素水平 ,行睾丸组织学检查 ,评估取精的成功率、受精率、种植率和临床妊娠率。　结果 :76例(76 % )经PESA获得精子 ,2 3例 (2 3% )通过TESE获得精子。PESA和TESE组的受精率、种植率和临床妊娠率分别为 71.3%和 75 .18% ,2 0 .35 %和 2 2 .0 5 % ,4 2 .11%和 4 1.6 0 %。PESA组有 32例临床妊娠 ,其中 15例继续妊娠 ,15例已分娩 ,2例流产。TESE组有 10例临床妊娠 ,其中 6例继续妊娠 ,2例已分娩 ,2例流产。两组的受精率、种植率和临床妊娠率差异无显著性。在TESE组有 1例取精失败而放弃治疗。　结论 :激素水平和睾丸组织学检查不能预测附睾或睾丸取精的成功 ,PESA和TESE获得精子进行单精子注射是治疗男性无精子症的有效方法 ,两组的受精率 ,种植率和临床妊娠率差异无显著性 (P >0 .0 5 )。     

Sperm source does not affect the ICSI outcome of patients with severely compromised spermatogenesis

Patients with spermatogenic dysfunction may display sperm parameters ranging from extremely severe oligozoospermia (sperm count lower than 2 million/ml) to azoospermia. It has been proposed that, since these patients may have increased sperm DNA damage that could affect their ICSI outcome, the use of surgically retrieved testicular spermatozoa should be preferred to improve their chance of fathering their biological offspring. However, studies in this field have yielded conflicting results. The present study provides an updated assessment of this subject by comparing the ICSI outcome of 762 patients with nonobstructive azoospermia and 419 with sperm count lower than 2 million/ml (median sperm count 300,000/ml). Both groups were homogeneous for the number of retrieved and injected MII oocytes. No difference was seen in terms of fertilisation, clinical pregnancy and cumulative live birth rates. Only the number of injected MII oocytes was found to independently predict the live birth rate, even when adjusted for the number of transferred embryos (OR 1.10 (1.0–1.2, p = 0.038)). The results of the present study stand against the use of testicular spermatozoa in patients with extremely severe spermatogenic dysfunction with available spermatozoa in their ejaculate.     

梗阻性无精子症患者睾丸抽吸精子行ICSI后的胚胎结局

目的总结非手术精子抽吸(NSA)联合ICSI对梗阻性无精子症患者的治疗意义,并探讨精子来源等对ICSI后胚胎结局的影响。方法回顾性分析642个ICSI周期,比较了睾丸抽吸精子和精液精子行ICSI后的受精率、优质胚胎率、临床妊娠率、种植率及胚胎停育率等。结果NSA睾丸精子组和精液精子组相比较,其2PN受精率、优质胚胎率、临床妊娠率、种植率及胚胎停育率之间均没有显著性差异(71．4％vs 73．2％, 73．1％vs 70．1％,34．2％vs 32．6％,22．3％vs 19．8％,21．3％vs 25．0％,P>0．05)。结论非手术抽吸睾丸精子联合ICSI是治疗梗阻性无精子症的一种有效方法。     

Comparison of pregnancy and neonatal outcomes of intracytoplasmic sperm injection performed with frozen versus fresh testicular sperm

BackgroundIt remains controversial whether there is a difference in the prognosis of intracytoplasmic sperm injection (ICSI) using frozen or fresh testicular sperm in patients with obstructive azoospermia (OA). Moreover, in the available studies, few have tracked neonatal outcomes. This study aimed to compare the pregnancy and neonatal outcomes of ICSI using cryopreserved sperm versus fresh sperm collected by testicular sperm aspiration (TESA).MethodsA total of 317 OA patients treated with ICSI in a university affiliated hospital from January 2016 to December 2020 were included in this study. The participants were divided into two groups according to the type of sperm used for ICSI: frozen sperm group (n=154) and fresh sperm group (n=163). The pregnancy and neonatal outcomes of the two groups were compared.ResultsThe data produced by this study showed no significant statistical difference in the 2 pronuclei (2PN) fertilization rate, 2PN cleavage rate, high-quality blastocyst rate, and the average number of transferred embryos in the frozen and fresh sperm groups. Similarly, no difference was found in implantation rate, clinical pregnancy rate, multiple pregnancy rate, miscarriage rate, premature delivery rate, live birth rate, and gender ratio at birth (P>0.05). The average newborn birth weight was similar in both groups (2,932.61±728.40 vs. 3,100.32±515.64 g, respectively) (P>0.05). A higher incidence of low birthweight (LBW) newborns was found in the frozen sperm group (20.91% vs. 8.49%) (P<0.05). Multiple logistic regression analysis showed that LBW is related to single or twin pregnancies (P<0.01), but not sperm (frozen or fresh) (P>0.05). We further analyzed the twin and single pregnancies in the two groups separately, and found that the incidences of LBW were both similar (P>0.05). There was no difference in the Apgar scores at 1 min and 5 min after birth between the two groups (P>0.05).ConclusionsThe use of frozen testicular sperm by TESA was efficient for men with OA. There were similar pregnancy and neonatal outcomes following TESA-ICSI using frozen or fresh sperm in this retrospective study. Prospective investigations are needed for further validation.     

Sperm retrieval and intracytoplasmic sperm injection outcomes with testicular sperm aspiration in men with severe oligozoospermia and cryptozoospermia

IntroductionSeveral studies addressed the role of testicular sperm aspiration with intracytoplasmic sperm injection (ICSI) in azoospermic men, but few have included non-azoospermic men. The aim of this study was to evaluate testicular sperm aspiration (TESA) sperm retrieval rates and ICSI outcomes in men with severe oligozoospermia.MethodsData were collected retrospectively from 88 consecutive, non-azoospermic, infertile men with idiopathic severe oligozoospermia who underwent TESA between January 2011 and January 2018. Patients were categorized into four groups according to sperm concentration: <5 and >1 million/ml (group 1), <1 and > 0.1 million/ml (group 2), <0.1 million/ml (group 3), and cryptozoospermia (group 4).ResultsMean male age was 37±7 years and the mean female age was 33±4 years. Sperm was recovered successfully in 90% (79/88) of the men overall and in 100% (30/30) of the men in group 1, 97% (29/30) of the men in group 2, 88% (15/17) of the men in group 3, and 45% (5/11) of the men in group 4. Most (65%, 57/88) of the couples had an embryo transfer (ET). The overall clinical pregnancy rate per ET was 46% (26/57). The clinical pregnancy rates (per ET) were 43% (9/21) in group 1, 65% (13/20) in group 2, 36% (4/11) in group 3, and 0% (0/5) in group 4.ConclusionsOur data indicate TESA allows for high sperm retrieval rates and acceptable ICSI pregnancy rates in men with severe oligozoospermia. However, in our experience, TESA sperm retrieval rates and ICSI outcomes are poor in cryptozoospermic men.     

Outcome of in vitro fertilization and intracytoplasmic injection of epididymal and testicular sperm obtained from patients with obstructive and nonobstructive azoospermia

PURPOSE: We assessed fertilization, pregnancy and miscarriage rates in patients with obstructive and nonobstructive azoospermia who underwent intracytoplasmic sperm injection. MATERIALS AND METHODS: From June 1996 to March 2000, 166 consecutive patients (198 intracytoplasmic sperm injection cycles) with azoospermia were studied. Of these 198 cycles 68 were performed due to nonobstructive azoospermia using testicular spermatozoa and 130 were performed due to obstructive azoospermia using epididymal spermatozoa. RESULTS: The normal (2 pronuclei) and abnormal (1 plus 3 pronuclei) fertilization rates for obstructive and nonobstructive azoospermia were 60.5% and 16.6%, and 54% and 16.4%, respectively (p >0.05). The pregnancy rate per cycle, pregnancy rate per patient and abortion rate were 30%, 39.8% and 28% for obstructive azoospermia, and 22%, 28.3% and 40% for nonobstructive azoospermia (p <0.05). The normal and abnormal fertilization rates were 58.7% and 21.4% for percutaneous epididymal sperm aspiration (PESA), 62.3% and 10.4% for PESA plus testicular sperm aspiration (TESA), and 57.3% and 14.5% for TESA, respectively (p >0.05). The pregnancy rate per cycle, pregnancy rate per patient and abortion rate were 34.6%, 54.5% and 11.1% for PESA, 37.5%, 37.5% and 33.3% for PESA plus TESA, and 26.1%, 31% and 41% for TESA, respectively (PESA versus PESA plus TESA p >0.05, and PESA and PESA plus TESA versus TESA p <0.05). Epididymal or testicular motile sperm resulted in a lower abortion rate than epididymal or testicular immotile sperm (p = 0.03). CONCLUSIONS: No differences were noted in the fertilization and embryo transfer rates irrespective of etiology (obstructive versus nonobstructive) and type of spermatozoa (epididymal versus testicular). Testicular sperm retrieval results in lower fertilization and pregnancy rates as well as higher abortion rates than epididymal sperm retrieval.     

睾丸精子行ICSI改善严重畸形精子症患者治疗结局5例报告

目的:探讨利用睾丸精子行卵细胞胞质内单精子注射(ICSI)治疗严重畸形精子症患者(精液或附睾液精子畸形率≥99%)的可行性,改善辅助生殖技术治疗结局。方法:回顾性分析5例严重畸形症精子患者(附睾液精子,n=4;精液精子,n=1)利用不同来源精子行ICSI治疗的临床资料,并比较睾丸精子组与非睾丸精子组(附睾液精子和精液精子)之间受精率、卵裂率、优质胚胎率、妊娠率以及种植率的差异。结果:5例严重畸形精子症患者取精液精子或附睾液精子行ICSI治疗后无1例妊娠,而改用睾丸精子行ICSI治疗后4例成功妊娠。睾丸精子组与非睾丸精子组之间受精率、卵裂率及优质胚胎率均无显著差异(P>0.05),而睾丸精子组妊娠率和种植率均显著高于非睾丸精子组(P<0.01)。结论:对应用附睾精子或精液精子行ICSI治疗失败的严重畸形精子症患者改用睾丸精子治疗可有效改善其治疗结局。     

Fertilization of in vitro matured human oocytes by intracytoplasmic sperm injection （ICSI） using ejaculated and testicular spermatozoa

AIM: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). METHODS: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. RESULTS: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. CONCLUSION: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.     

Chromosomal and Y‐chromosome microdeletion analysis in 1,300 infertile males and the fertility outcome of patients with AZFc microdeletions

The present study investigated the frequency of chromosome aberrations and AZF microdeletions in infertile patients with nonobstructive azoospermia (NOA) or severe oligozoospermia. Additionally, the effect of the AZFc microdeletions on the success of microdissection testicular sperm extraction (microTESE) and intracytoplasmic sperm injection (ICSI) methods were evaluated. Peripheral blood samples were received from 1,300 infertile men with NOA and severe oligozoospermia. Karyotyping and FISH analysis were performed according to standard methods. AZF microdeletions were analysed using multiplex polymerase chain reaction or GML Y‐chromosome Microdeletion Detection System consisting of 14 markers. The chromosomal aberrations and the AZF microdeletions frequency among 1,300 infertile men were 10.6% and 4.0% respectively. Either ejaculated spermatozoa or microTESE was performed on only in 19 out of 26 patients with AZFc deletions. Of the 19 patients, four had severe oligozoospermia and 15 had NOA. In eight out of 15 NOA patients, testicular mature spermatozoa were obtained (53.3%) and then ICSI was applied to mature oocytes. After undergoing ICSI treatment, clinical pregnancy and live birth outcome rates were found to be 37.5% and 25% respectively. These results suggest that infertile patients with AZFc microdeletion could achieve successful fertilisation pregnancies with the help of assisted reproductive technology.     

未处理或上游法处理后精子的DNA碎片能否预测使用赠卵ICSI后的妊娠？

本研究比较了未处理的精子和上游法处理后精子中的DNA碎片值（SDF）预测使用赠卵ICSI后妊娠的可能性。总共81例不孕女性接受通过夫精和赠卵ICSI后的胚胎移植。这种方式排除了目前公认的影响精子DNA修复的女方因素。在ICSI受精时，同时使用DNA碎片率检测试剂盒（Halosperm）进行盲法检测未处理的和上游法处理后精液中的精子DNA碎片。经上游法处理后的精液样本中SDF值呈显著下降。有趣的是，未处理的或上游法处理后精液样本中的SDF值均能预测妊娠。通过ROC曲线和其衍生的约登指数计算得出未处理前和上游法处理后精子sDF的临界值分别是24．8％和17．5％。未处理精子SDF预测妊娠的敏感性和特异性分别为75％和69％，而上游法处理后的精子SDF预测妊娠的敏感性和特异性分别是78％和73％。虽然SDF值增高与生育呈负相关，但我们发现在精子筛选降低sDF值后再与赠卵进行ICSI并不能保证获得妊娠。这表明目前的技术手段对于一些DNA损伤仍无法探及，从而相对影响辅助生殖成功的机率。因此，我们认为对目前的“冰山模式”作一些修饰后可以用来解释SDF在生育中的作用。     

ICSI中不同来源精子对临床结局的影响

目的:分析不同来源精子在卵细胞胞质内单精子注射(ICSI)后的临床结局。方法:将682个ICSI治疗周期分为:射出精子组(598例)、经皮附睾精子抽吸术(PESA)得到精子组(58例)、睾丸精子获取术(TSE)得到精子组(26例),比较三组的受精率、临床妊娠率、种植率、流产率、异位妊娠率以及分娩率之间的差别。结果:TSE组受精率明显低于射出精子组和PESA组(81.06%vs87.95%,87.82%,P<0.05);射出精子组、PESA组和TSE组的妊娠率(39.46%,48.28%,34.62%)、种植率(19.80%,23.80%,18.34%)、流产率(13.13%,17.86%,11.11%)、异位妊娠率(5.51%,7.14%,11.11%)、分娩率(32.11%,36.21%,26.92%),均没有显著差异(P>0.05)。结论:虽然TSE来源精子对ICSI受精率有所影响,但射出精子和手术(TSE和PESA)来源精子对妊娠结局没有影响。