精子冷冻对卵胞质内单精子注射治疗结果的影响

目的 了解冷冻保存对射出精子和经皮附睾、睾丸微穿刺抽吸精子进行卵胞质内单精子注射（ICSI）临床结果的影响。方法 将射出精子152个治疗周期和经皮附睾、睾丸微穿刺抽吸精子55个周期分别分为新鲜组和冷冻组，对其ICSI后的结果进行对照。结果 射出精子新鲜组和冷冻组受精率、卵裂率和临床妊娠率（分别为79．70％ vs 76．79％、97．08％ vs 98．35％、39．02％ vs 37．93％）与经皮附睾、睾丸微穿刺抽吸精子新鲜组和冷冻组受精率、卵裂率和临床妊娠率，穿刺周期（分别为77．07％ vs 74．54％、97．47％ vs 94．3l％、35．00％ vs 37．14％）均无显著差异（P〉0．05）。结论 精子的冷冻保存对射出精子和经皮附睾、睾丸微穿刺抽吸精子进行ICSI的临床结果没有影响，我们认为对于经皮附睾、睾丸微穿刺抽吸精子应该进行冷冻保存，以减少对附睾、睾丸的穿刺次数。     

睾丸精子辅助生育30例报告

本生殖中心自 1 997年至今 ,用睾丸精子卵浆内单精子注射 (intracytoplasmic sperm injec-tion,ICSI)为 30对无精子症不育夫妇行辅助生育治疗 32个周期 ,报道如下。一、资料与方法30例无精子症不育、染色体核型正常的患者平均年龄 35 (2 2～ 45 )岁 ,治疗前均检查血卵泡刺激素 (FSH)、黄体生成素 (L H)、睾酮 (T)、泌乳素 (PRL )水平及睾丸活检了解睾丸功能及生精状况。首先对女方促超排卵治疗 [1] ,女方平均年龄 31 (2 1～ 39)岁 ,当卵泡发育成熟时 ,以阴道B超介导下取卵 ,卵子的处理及 ICSI方法见文献 [2 ]。获得卵子后即行睾丸精…     

睾丸精子获取和单精子卵胞浆内注射术治疗非梗阻性无精子症不育

目的　探讨非梗阻性无精子症患者外科获取睾丸精子的方法和意义。　方法　 4 9例非梗阻性无精子症患者行开放睾丸活检和诊断性睾丸精子获取术 (TESE) ,诊断性TESE有精子者至少 3个月后行单精子卵胞浆内注射 (ICSI)治疗。　结果　 12例 (2 4 .9% )诊断性TESE中发现精子 ,其中 3例为生精减少 ,2例为生精阻滞 ,7例为Sertoli细胞综合征。睾丸体积、血FSH水平和睾丸病理类型不能准确预测是否有精子。 8例行ICSI治疗 ,7例 (87.5 % )再次TESE获得睾丸精子行显微注射 ,3例获得临床妊娠。　结论　非梗阻性无精子症患者有必要行诊断性TESE确定睾丸内是否存在精子 ,获取睾丸精子结合ICSI可以有效治疗非梗阻性无精子症不育。     

冷冻睾丸精子行卵泡浆内单精子注射术的结局评估

目的 梗阻性无精子症患者采用诊断性睾丸精子抽吸术(TESA),对获得的睾丸精子进行冷冻,评估新鲜精子和冷冻精子分别行卵泡浆内单精子注射术(ICSI)的受精率和妊娠率. 方法 回顾分析本院2007～2010年就诊的237对梗阻性无精子症(OA)夫妇用新鲜睾丸精子行ICSI和2010～2011年就诊的31例OA患者的冷冻睾丸精子行ICSI后胚胎移植的资料. 结果 新鲜精子组和冷冻精子组的平均成熟卵泡数分别为(14.13±7.41)和(14.31±8.14),平均正常受精率分别为82.99％和84.17％,种植率分别为24.08％和25.64％,以上指标两组相比较均无显著性差异(P＞0.05).两组的临床妊娠率也没有显著差异(新鲜精子组43.88％和冷冻精子组48.39％,P＞0.05). 结论 冷冻睾丸精子不影响ICSI的受精率和临床妊娠率,采用TESA获取的冷冻睾丸精子行ICSI是一种能减少患者痛苦又安全、有效的方法.     

梗阻性无精子症经皮睾丸穿刺行卵胞浆内单精子注射治疗44例

已婚夫妇中约有10％～15％不育，其中男性因素占30％～50％，梗阻性无精子症占无精子症的50％以上。1992年Palermo等首次成功使用卵胞浆内单精子注射（ICSI）获得妊娠，使无精子症男性能够获得自己的后代。Yemini等报道对无精子症患者进行睾丸切开活检（TESE）后行ICSI治疗获妊娠，但开放性的睾丸活检术特别是重复开放性活检，可能导致出血、感染，甚至永久性睾丸缺血。对于局灶性生精功能下降引起的睾丸功能衰竭，开放手术又局限了活检部位，不能全面反映睾丸的生精功能。     

睾丸细针抽吸取精行ICSI治疗阻塞性无精子症

目的 :探讨睾丸细针抽吸技术 (TEFNA)和睾丸精子行卵胞浆内单精子注射 (ICSI)治疗阻塞性无精子症(OAS)的可行性和效果。　方法 :对 14对因男性OAS引起不育的夫妇进行 15个周期的TEFNA +ICSI治疗 ,女方常规促超排卵 ,在取卵当日从男方睾丸用细针抽吸精曲小管 ,经体外处理、培养后 ,选择精子行ICSI,其中 14个周期使用活动精子 ,1例病人同时采用附睾精子和睾丸精子。　结果 :共获卵子 15 2个 ,成熟卵 132个 ,受精卵 82个 (6 2 .1% ) ;共行胚胎移植 14个周期 ,平均移植胚胎 2 .5个 ,临床妊娠 11例。　结论 :TEFNA易操作 ,创伤小 ,并发症少 ;从睾丸组织中分离活动精子行ICSI是治疗男性无精子症的一种有效方法。     

完全不运动精子经冻融后应用激光筛选存活精子行ICSI获临床妊娠

目的报道1例睾丸完全不运动精子经冷冻-解冻后应用激光筛选存活精子行卵胞浆内单精子注射(ICSI)获得临床妊娠。方法睾丸穿刺获取的完全不运动精子经激光鉴定有存活的精子,立即进行冷冻保存。取卵日,解冻该精子应用激光选择存活的精子进行卵胞浆内单精子注射。结果睾丸完全不运动精子冷冻前和解冻后经激光鉴定存活率分别为55.75%和39.81%。选择激光鉴定为存活的精子注射到5个成熟卵母细胞中,4个卵子正常受精(受精率80%),培养至第3天均卵裂(卵裂率100%),移植两枚优质胚胎后获得临床妊娠。结论完全不运动精子是可以冷冻保存的。     

不同来源的精子ICSI治疗周期妊娠结局分析

目的:比较不同来源的精子进行ICSI治疗后受精率、胚胎种植率、临床妊娠率等临床指标有无差异。方法:回顾性分析2006年1月~2008年12月本院生殖中心进行的431个ICSI治疗周期,按精子来源分为A组(重度少弱精子症组)287个周期、B组(梗阻性无精子症附睾穿刺组)109个周期、C组(梗阻性无精子症睾丸穿刺组)35个周期,比较各组女方平均年龄、男方平均年龄、不孕病史、平均MII卵数、受精率、卵裂率、胚胎利用率、平均移植胚胎数量、种植率、妊娠率、流产率等指标的差异。结果:A组与B、C两组在种植率、妊娠率方面差异有统计学意义(18.46%vs25.23%、28.76%;31.23%vs42.16%、39.39%,P<0.05);B、C两组之间各数据差异无统计学意义(P>0.05),受精率、卵裂率、流产率3组之间差异无统计学意义。结论:重度少弱精子症患者射出精子进行ICSI治疗后胚胎种植率、临床妊娠率低于梗阻性无精子症患者。     

146例炎症梗阻性无精子症的临床评估和ICSI治疗结局分析

目的分析炎症梗阻性无精子症的临床评估和单精子卵胞浆内注射(ICSI)的治疗结局。方法前瞻性研究近5年间接受ICSI治疗的炎症性梗阻性无精子症的临床特征、精液和超声特点,经皮附睾穿刺精子抽吸术(PESA)或经皮睾丸穿刺取精术(TEFNA)结合ICSI治疗后观察受精、临床妊娠等结果。结果146例患者体检附睾均有增粗变硬或伴头尾部结节。82例患者曾有生育史、附睾炎症史或输精管附睾吻合手术史,其中72例PESA找到附睾精子;53例无上述病史者49例PESA找到附睾精子:另有精道远端梗阻11例。ICSI治疗146例167周期炎症性梗阻性无精子症的受精率、每周期临床妊娠率分别为81.1%和42.1%。结论炎症梗阻性无精子症具备典型的临床和超声特征,PESA附睾精子获取率高,ICSI治疗获得较高受精率和临床妊娠率。     

经皮睾丸精子抽吸术的临床应用

目的 探讨经皮睾丸精子抽吸术在临床上的应用。方法 135例患者分为3组:梗阻性无精子症组(OA组)71例,先天性无精子症组(CC组)23例,原发性无精子症组(IA组)41例,所有患者均做经皮睾丸精子抽吸术及单精子注射术。结果 通过经皮睾丸精子抽吸术(PTAS)获得精子的平均密度为73.542.59/μl,平均活动率为(1.51.17)%(范围1%~5%),共1263个成熟卵子行ICSI,获得849个正常胚胎(正常受精率为67.2%)。在OA组中,71例患者行79周期,32例怀孕。在CC组中,23例患者行28周期,9例怀孕。在IA组中,41例患者行41周期,8例怀孕。结论 PTSA合ICSI是一种治疗梗阻性无精子症的有效手段,且梗阻性无精子症患者的妊娠率高于特发性无精子症患者。     

Testicular fine needle aspiration for sperm retrieval in non-obstructive azoospermia

Objective: To investigate the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by testicular fine needle aspiration (TEFNA) in men diagnosed non-obstructive azoospermia. Methods: TEFNA was performed in 121 patients with a mean of 15 punctures and aspirations from each testis with a #23 butterfly needle connected to a 20 mL syringe with an aspiration handle. Results: One hundred and twenty-one patients underwent 176 TEFNA cycles. Testicular sperm were recovered in 56.3 % (99/176) cycles from 57 % (69/121) of patients. The sperm recovery rate was 46.7 % (21/45) in patients with Sertoli cell-only syndrome, 45.7 % (16/35) in patients with maturation arrest, 96.1 % (25/26) in patients with hypospermatogenesis and 63.6 % (7/11) in patients of non-mosaic Klinefelter's syndrome as judged by testicular histology. No sperm were found in 3 cases with post-irradiation fibrosis and one, after resection and chemotherapy of unilateral testicular cancer. In 87 cycles of ICSI using     

睾丸精子细针抽吸结合ICSI治疗非阻塞性无精子症

目的探讨应用睾丸细针穿刺(TEFNA)在非阻塞性无精子症患者获取成熟精子,结合ICSI治疗男性不育。方法用23号蝶型针与20ml注射器相连,注射器固定于保持其负压的手柄上,平均每个睾丸穿刺15针。结果57%(69/121)的患者,56.3%(99/176)个周期获得成熟精子,经睾丸组织学检查不同类型患者的精子获取率分别为:唯支持细胞46.7%(21/45),精子成熟障碍45.7%(16/35),精子发生低下96.1%(25/26),非嵌合型Klinefelter综合征63.6%(7/11),3例放疗后睾丸纤维化及1例单侧睾丸癌切除化疗后未获得精子。在仅注射了丈夫精子的87个周期中,591个卵细胞行精子注射,其中36.9%(218/591)受精,得到202个胚胎,其62个周期移值了178个胚胎(每个周期平均为2.831.7个胚胎),得到26例临床妊娠(共有44个妊娠囊),妊娠率为41.9%,种植率为24.7%,结论TEFNA结合ICIS是一种简易、安全、有效、易被患者接受的治疗非阻塞性无精子症的方法。     

Surgical recovery of sperm in non-obstructive azoospermia

The development of intracytoplasmic sperm injection (ICSI) opened a new era in the field of assisted reproduction and revolutionized the assisted reproductive technology protocols for couples with male factor infertility. Fertilisation and pregnancies can be achieved with spermatozoa recovered not only from the ejaculate but also from the seminiferous tubules. The most common methods for retrieving testicular sperm in non-obstructive azoospermia (NOA) are testicular sperm aspiration (TESA: needle/fine needle aspiration) and open testicular biopsy (testicular sperm extraction: TESE). The optimal technique for sperm extraction should be minimally invasive and avoid destruction of testicular function, without compromising the chance to retrieve adequate numbers of spermatozoa to perform ICSI. Microdissection TESE (micro-TESE), performed with an operative microscope, is widely considered to be the best method for sperm retrieval in NOA, as larger and opaque tubules, presumably with active spermatogenesis, can be directly identified, resulting in higher spermatozoa retrieval rates with minimal tissue loss and low postoperative complications. Micro-TESE, in combination with ICSI, is applicable in all cases of NOA, including Klinefelter syndrome (KS). The outcomes of surgical sperm retrieval, primarily in NOA patients with elevated serum follicle-stimulating hormone (FSH) (NOA including KS patients), are reviewed along with the phenotypic features. The predictive factors for surgical sperm retrieval and outcomes of treatment were analysed. Finally, the short- and long-term complications in micro-TESE in both 46XY males with NOA and KS patients are considered.     

Efficiency of percutaneous testicular sperm aspiration as a mode of sperm collection for intracytoplasmic sperm injection in nonobstructive azoospermia

PURPOSE: We determined the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by percutaneous testicular sperm aspiration in men with nonobstructive azoospermia. We also compared the results of ICSI using spermatozoa recovered by open excisional biopsy versus percutaneous testicular sperm aspiration. MATERIALS AND METHODS: A total of 84 men with nonobstructive azoospermia underwent percutaneous testicular sperm aspiration to recover testicular spermatozoa for ICSI on the day of ova retrieval from the wife. Percutaneous testicular sperm aspiration was performed with the patient under general anesthesia in the upper and lower poles of each testis. It was followed by immediate microscopic search of the aspirate to confirm the presence of spermatozoa. In the absence of spermatozoa open excisional biopsy was performed in the same setting. RESULTS: Percutaneous testicular sperm aspiration resulted in the recovery of mature spermatozoa in 45 men (53.6%). Of the remaining 39 men (46.4%) requiring open biopsy adequate spermatozoa were recovered in 28 (71.8%). Although the fertilization rate was significantly higher in the sperm aspiration group, the cleavage and pregnancy rates were similar in the 2 groups. CONCLUSIONS: Percutaneous testicular sperm aspiration was a successful initial approach to collect mature spermatozoa in a high proportion of men with nonobstructive azoospermia. It is safe, minimally invasive and well tolerated by all patients.     

Testicular fine needle aspiration as a diagnostic tool in non-obstructive azoospermia

Aim： To report the fine needle aspiration cytology （FNAC） of the testes used as a diagnostic tool in non-obstructive azoospermic patients. Methods： One hundred and twenty-five non-obstructive azoospermic male candidates to intracytoplasmic sperm injetion （ICSI） were analysed for follicle stimulating hormone （FSH）, luteinizing hormone （LH）, testosterone and inhibin B plasma levels. They were classified into three groups on the basis of FNAC： 1） Sertoli cell-only syndrome （SCOS） （70）; 2） severe hypospermatogenesis （42）; and 3） maturation arrest （13）, Then, all men underwent testicular sperm extraction （TESE） for sperm recovery for ICSI. Results： Mature spermatozoa were detected by FNAC in 24 of 42 men with severe hypospermatogenesis and nine of 13 men with maturation arrest; while they were retrieved by TESE in 29 of 70 men with SCOS, 35 of 42 men with severe hypospermatogenesis （including the 24 by FNAC） and 10 of 13 men with maturation arrest （including the nine by FNAC）. The sensitivity and specificity of FNAC were 44.6 % and 100 %, respectively. There was no difference on testicular volume and hormonal parameters in men with and without sperm retrieved. Conclusion： These findings suggest that FNAC may be a simple and valid diagnostic parameter in non-obstructive azoospermic men and it may represent a valid positive prognostic parameter for sperm recovery at TESE, （Asian J Androl 2005 Sep; 7： 289-294）     

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.     

Intracytoplasmic sperm injection using surgically retrieved epididymal and testicular spermatozoa in cases of obstructive and non-obstructive azoospermia

This was a retrospective study of 115 patients who underwent 124 cycles of ICSI using surgically retrieved spermatozoa. The objective was to compare the results of ICSI in patients with obstructive azoospermia using epididymal spermatozoa (36 cycles) or testicular spermatozoa (58 cycles) with ICSI in patients with non-obstructive azoospermia using testicular spermatozoa (30 cycles). When epididymal spermatozoa were used for ICSI, the fertilization rate per injected metaphase-II oocyte and the clinical pregnancy rate per ICSI cycle were 60.4 and 25%, respectively. When testicular spermatozoa were used in obstructive cases, the fertilization rate and pregnancy rate were 57.9 and 34.5%. In non-obstructive cases the fertilization and pregnancy rates were 41.2 and 16.6%. When patients with obstructive azoospermia were regrouped according to the cause of obstruction, the fertilization and pregnancy rates were 59.1 and 35.1% in acquired obstruction and 58.7 and 24.3% in congenital obstruction. The fertilization and pregnancy rates were not statistically different ( p  > 0.05) when testicular or epididymal spermatozoa were used in obstructive cases; neither was statistically different ( p  > 0.05) when compared in patients with congenital and acquired obstruction. On the other hand, the fertilization and pregnancy rates in cases with non-obstructive azoospermia were significantly lower ( p  < 0.05) than in obstructive cases.     

Sperm retrieval procedures and intracytoplasmatic spermatozoa injection with epididymal and testicular sperms

INTRODUCTION: Male infertility caused by azoospermia due to non-reconstructable obstruction or non-obstructive azoospermia can be treated by microsurgical epididymal aspiration (MESA) or testicular sperm extraction (TESE) followed by an intracytoplasmatic spermatozoa injection (ICSI). MATERIAL AND METHODS: From 9/93 to 6/01, we carried out 1,025 ICSI procedures with aspirated epididymal or testicular sperms in 684 cases. 163 ICSI cycles were performed with epididymal sperms and 862 ICSI cycles with testicular sperms or spermatids. The TESE was carried out by open biopsy, frequently in a multilocular technique. The aspirated spermatozoas were used after cryopreservation (frozen) or immediately after aspiration (fresh). RESULTS: 538 patients had obstructive azoospermia or ejaculation failure. In 487 cases the underlying cause of azoospermia was an impaired spermatogenesis, following maldescensus testis, chemotherapy, radiotherapy, or caused by Sertoli-cell-only syndrome, a genetic disorder or an unknown etiology. The transfer rates, pregnancy rates and birth rates per ICSI cycle showed no statistically significant differences between testicular and epididymal sperms in the cases of seminal obstruction (28% average birth rates in both cases). However, highly significant was the difference in birth rates with regard to the underlying cause of infertility. In contrast, in treating non-obstructive azoospermia we observed a birth rate of 19% per cycle. In all patient groups the birth rate with fresh spermatozoas did not differ from those with cryopreserved spermatozoa. 40% of patients after multilocular TESE showed clinical signs of testicular lesion. CONCLUSION: The underlying cause of azoospermia is the most important factor for the outcome of ICSI using epididymal and testicular sperms. In cases of non-obstructive azoospermia, the pregnancy rate is low compared with the results in cases of obstructive azoospermia. There is no difference between fresh and cryopreserved sperms. TESE with ICSI is the most efficient treatment of azoospermia caused by hypergonadotropic hypogonadism. The morbidity of the TESE procedure is highly relevant and must be considered if this technique is indicated.