经皮附睾或睾丸抽吸取精结合ICSI治疗无精子症

目的评价经皮附睾精子抽吸术（percutaneus epididymal sperm aspiration,PESA）或睾丸精子抽吸术（testicu1ar sperm aspiration,TESA）结合卵胞浆内单精子注射（intracytoplasmic sperm injection,ICSI）治疗无精子症的临床效果。方法对290例因男性梗阻性及非梗阻性无精子症（non-obstructive azoospermia,NOA）采用PESA或TESA穿刺获取精子,女方采用长方案超排卵,然后对处于细胞分裂中期的成熟卵母细胞进行单精予注射。结果梗阻性无精子症组203例,受精率77．5％,临床妊娠率46．1％;非梗阻性无精子症组87例,受精率73．O％,临床妊娠率41．4％,两组比较其受精率及临床妊娠率均无显著性差异（P〉0．05）。结论采用PESA或TESA获取精子结合ICSI是治疗梗阻性及非梗阻性无精子症等严重的男性不育症的一种有效的方法。     

睾丸与附睾精子对梗阻性无精子症ICSI助孕结局的影响

目的比较睾丸精子与附睾精子对梗阻性无精子症(obstructive azoospermia,OA)患者行卵胞浆内单精子注射(intracytoplasmic sperm injection,ICSI)治疗结局的影响。方法收集2014年1月至2016年12月在海南医学院第一附属医院生殖医学中心因梗阻性无精子症行ICSI助孕治疗患者的临床资料,共163个周期。根据精子来源分为两组,TESA组:采用睾丸精子抽吸术(testicular sperm aspiration,TESA)取精,共137个周期;PESA组:采用经皮附睾精子抽吸术(percutaneous epididymal sperm aspiration,PESA)取精,共26个周期。比较两组的正常受精率、2PN卵裂率、优质胚胎率、胚胎利用率、胚胎种植率、临床妊娠率、流产率及活产率之间有无差异。结果 TESA组的胚胎种植率、临床妊娠率及活产率高于PESA组,但差异均无统计学意义(P0.05);TESA组的正常受精率、2PN卵裂率、优质胚胎率、胚胎利用率及流产率均低于PESA组,但差异亦无统计学意义(P0.05)。结论对梗阻性无精子症患者,睾丸精子和附睾精子ICSI后可达到相似的助孕结局。     

经皮睾丸精子抽吸术治疗无精子症的研究

目的探讨经皮睾丸精子抽吸术(PTSA)获取睾丸精子结合卵胞浆内单精子注射术(ICSI)治疗梗阻性和非梗阻性无精子症,使之获得亲生子女.方法对121例因男性梗阻性及非梗阻性无精子症患者进行诊断性穿刺,均证实有精子后进行119个周期PTSA ICSI治疗.结果共获卵子1514个,成熟卵985个,胚胎741个,平均每例6.23个胚胎,总受精率74.4%,卵裂率97.6%;共移植114个周期和冷冻胚胎移植5个周期,平均移植2.86个胚胎,B超证实临床妊娠48例,临床妊娠率40.3%.结论采用PTSA技术获取的睾丸精子进行ICSI是治疗梗阻性及非梗阻性无精子症的一种安全、简单、有效的方法.     

新鲜附睾精子与冷冻复苏后的附睾精子临床结局比较

目的探讨经皮附睾穿刺取精术（PESA）获得精子经冷冻复苏后行卵胞浆内单精子注射术（ICSI）的临床效果。方法将采用新鲜附睾精子的94例患者作为新鲜组对照,冷冻复苏后附睾精子的92例患者作为冻精组,比较二组的受精率,可利用胚胎率及妊娠率。结果新鲜组与冻精组相比受精率,可利用胚胎率及妊娠率无显著性差异（75.04%vs78.4%,78.3%vs81.3%,47.87%vs44.44%P〉0.05）。结论PESA精子经冷冻后有很好的复苏率（97.94%）,结合ICSI可得到与使用新鲜PESA精子同样的临床结局,可作为梗阻性无精症的治疗方法。     

附睾精子和睾丸精子妊娠结局比较

目的比较经皮附睾穿刺抽吸术和经皮睾丸精子抽吸术两种方法获得的精子妊娠结局。方法83例无精子症患者经皮附睾穿刺抽吸术取得附睾精子；35例无精子症患者经皮睾丸精子抽吸术（TESA）获得睾丸精子。女方进行常规超排卵。采用卵胞浆内单精子注射技术获得妊娠，比较两者的受精率、种植率和临床妊娠率。结果附睾精子组和睾丸精子组的受精率分别为75．20％和74．61％，比较其差异无显著性（P〉0．05）；两者的种植率和临床妊娠率分别为29．18％VS23．89％和52．43％VS40．21％，差异具有显著性（P〈0．05）。结论附睾是精子获能、成熟的重要部位，附睾精子优于睾丸精子，对无精子症患者行ICSI之前尽可能首先选取附睾精子。     

睾丸精子与附睾精子结合单精子注射治疗无精子症妊娠结局比较

目的比较附睾精子和睾丸精子结合单精子注射(ICSI)治疗无精子症的妊娠结局。方法 216例无精子症患者经皮附睾穿刺抽吸术取得附睾精子;74例无精子症患者经皮睾丸精子抽吸术(TESA)获得睾丸精子。女方进行常规超排卵。采用卵胞浆内单精子注射技术获得妊娠,比较两者的受精率、种植率和临床妊娠率。结果附睾精子组和睾丸精子组的受精率分别为75.20%和74.61%,比较其差异无显著性(P0.05);两者的种植率和临床妊娠率分别为29.18%vs 23.89%和52.43%vs 40.21%,差异具有显著性(P0.05)。结论附睾是精子获能、成熟的重要部位,附睾精子优于睾丸精子,对无精子症患者行ICSI之前尽可能首先选取附睾精子。     

附睾穿刺取精行ICSI治疗梗阻性无精子症

目的探讨附睾穿刺取精术(PESA)结合单精子卵胞浆内注射(ICSI)治疗梗阻性无精子症男性不育的可行性,并观察其临床效果。方法 7对夫妇为研究对象,男方均确诊为梗阻性无精子症,女方超促排卵获得卵细胞,男方于取卵日在局麻下通过细针穿刺附睾头部吸取少量精子,行ICSI,受精成功后24-48h,选择优质胚胎移植入子宫腔。因男性少弱精子症行ICSI治疗的20个治疗周期为对照组。结果附睾取精7例共11个治疗周期全部获得活动精子,ICSI后受精率65．9％,卵裂率98．3％,优质胚胎率71．9％,临床妊娠5例,周期临床妊娠率45．5％,与对照组比较,各项指标均无显著差异。结论附睾穿刺取得的精子与排出体外的精子具有相同的受精和获得优质胚胎的能力,PESA是治疗梗阻性无精子症男性不育的安全有效的方法。     

睾丸容积、染色体及血清生殖激素对无精子症睾丸内精子存在的评估作用

无精子症,在男性不育患者中占5%～20%,包括染色体畸变、内分泌激素异常等病因[1]。无精子症病因复杂、临床治疗困难,是导致男性绝对不育的重要原因。临床上常采用经皮附睾精子抽吸术（percutaneous epididymal sperm aspira-tion,PESA）和睾丸精子抽吸术（testicular sperm extraction,TESE）,通过手术获取成熟精子行辅助生殖技术。     

冷冻睾丸精子行卵胞浆内单精子注射临床结局探讨

目的探讨冷冻睾丸精子行卵胞浆内单精子注射(ICSI)对无精子症患者临床结局的影响。方法回顾性分析2017年1月至2018年12月在我院生殖中心经皮睾丸穿刺取精(TESA)行ICSI助孕的213个授精周期的临床资料。按精子来源分为两组,TESA冻精组(观察组):采用TESA冷冻精子,共114个周期;TESA组(对照组):采用新鲜TESA精子,共99个周期。比较两组的胚胎发育情况和临床结局。结果 TESA冻精组正常受精率(2PN率)显著低于TESA组,差异具有统计学意义(64.26% vs 73.89%,P0.01)。两组的2PN卵裂率、可用胚胎率、优质胚胎率和囊胚形成率差别无显著性(P0.05)。两组的临床妊娠率、胚胎种植率、多胎妊娠率、异位妊娠率、早期流产率和分娩率差异没有统计学意义(P0.05)。结论冷冻睾丸精子行ICSI的临床结局与新鲜睾丸精子总体相似。但需要注意冷冻睾丸精子可能降低ICSI后正常受精率。     

睾丸精子冻融后行卵胞浆内单精子注射术在无精子症患者中的临床应用

目的探讨对无精子症患者实施睾丸精子经冷冻复苏后行卵胞浆内单精子注射术（ICSI）的临床效果。方法回顾性分析了既往在我中心有冷冻睾丸精子的34个ICSI周期,评估其冷冻精子复苏、卵子受精、卵裂、可移植胚胎、优质胚胎、临床妊娠及其分娩情况。结果 34个拟定复苏冷冻睾丸精子的周期精子复苏均获得成功,受精率为76.9%（249/324）,2PN受精率69.7%（226/324）,卵裂率98.8%（246/249）,可利用胚胎率84.2%（207/246）,优质胚胎率46.3%（114/246）;所有周期均有胚胎移植;34个周期中行新鲜胚胎移植30个,种植率为34.4%（21/61）,临床妊娠17例（临床妊娠率56.7%）包括13例单胎和4例双胎,其中1例单胎妊娠流产（流产率为5.9%）。目前,有14个周期已经出生了18个健康婴儿（10个男婴,8个女婴）未发现先天缺陷儿,另2例单胎继续妊娠中（32＋w,29＋w）。结论睾丸精子经冷冻后有很好的复苏率,结合ICSI受精可以得到较好的临床结局。     

Diagnostic epididymal and testicular sperm recovery and genetic aspects in azoospermic men.

Various procedures for sperm recovery in azoospermic men have been described, from open testicular biopsy to simple needle aspiration from the epididymis and the testis. Fifty-one obstructive and 86 non-obstructive azoospermic men were treated to compare the recovery of spermatozoa obtained by percutaneous aspiration from the epididymis (PESA) and aspiration/extraction from the testis (TESA, TESE) with histopathology. If TESA failed, the work up proceeded with TESE. All patients were karyotyped. Spermatozoa were recovered by PESA or TESA in all obstructive men (51/51 patients). In 22 out of 86 patients with non-obstructive azoospermia, testicular spermatozoa could be successfully recovered by TESA. In five additional patients TESE was successful in recovering spermatozoa where TESA had failed. In 43 patients, neither TESA nor TESE was successful. Sixteen patients chose not to proceed with TESE. Seven out of 86 patients had an abnormal karyotype in the non-obstructive group (8%), none in the obstructive group. In the non-obstructive patient group testicular histopathology showed hypospermatogenesis, incomplete maturation arrest and germ cell aplasia with focal spermatogenesis in cases where spermatozoa were recovered and complete germ cell aplasia, complete maturation arrest and fibrosis in cases where no spermatozoa were found. Spermatozoa were recovered by PESA or TESA from all patients with obstructive azoospermia and from approximately 40% of patients with non-obstructive azoospermia by TESA or TESE. Retrieval of viable spermatozoa in the infertility work-up was highly predictable for sperm recovery in subsequent ICSI cycles. TESA performed under local anaesthesia seems almost as effective as more invasive procedures in recovering testicular spermatozoa, both in obstructive and non-obstructive azoospermic men.     

Sperm retrieval and fertilization in repeated percutaneous epididymal sperm aspiration

Percutaneous epididymal sperm aspiration (PESA) for retrievalof spermatozoa for intracytoplasmic sperm injection (ICSI) isa new simplified technique in the treatment of men with obstructiveazoospermia. There has been a fear that the PESA procedure,being blind, could cause damage to the epididymal duct systemand make it impossible to retrieve spermatozoa if a repeatedprocedure is required. We report here on repeated PESA proceduresfrom the same unilateral epididymis. Twenty-seven men with obstructiveazoospermia were investigated retrospectively regarding sufficiencyof the number of motile spermatozoa for ICSI, fertilizationrate (FR) and possibility of collecting spermatozoa for cryopreservationin repeated PESA procedures. Sufficient motile spermatozoa forICSI were found in a similar proportion of men at the firsttwo attempts: 91 and 89% respectively. Fertilization rate andthe possibility of collecting spermatozoa for cryo-preservationwere also similar at the first two PESA procedures: 62 versus67% and 33 versus 33% respectively. At the third procedure,motile spermatozoa for ICSI were retrieved in 86% (6/7), FRwas 47% and spermatozoa were cryopreserved in one case. Twomen underwent a fourth PESA. In both cases, a sufficient numberof motile spermatozoa for ICSI was found and FR was 62%. Thisstudy shows that in men with obstructive azoospermia, PESA canbe repeated on the same unilateral epididymis up to three times,with good opportunity of retrieving sufficient motile spermatozoafor ICSI.     

High implantation and pregnancy rates with testicular sperm extraction and intracytoplasmic sperm injection in obstructive and non-obstructive azoospermia

Thirty-two infertile couples with obstructive and non-obstructiveazoospermia were included in this study. Testicular sperm extraction(TESE) was performed in 16 obstructive azoospermic cases wheremicrosurgical sperm aspiration (MESA) or percutaneous spermaspiration (PESA) were impossible because of totally destroyedepididymis and 16 non-obstructive azoospermia cases with severespermatogenetic defect where the testicles were the only sourceof sperm cells. A total of 288 oocytes was obtained from 32females and 84% were injected. The fertilization rates (FR)with 2 pronuclei (PN) and cleavage rate were 50.8 and 68.2%respectively. A total of 15 pregnancies was achieved (53% perembryo transfer), nine from the obstructive and six from thenon-obstructive group. Four pregnancies resulted in clinicalabortion (26.6%). The ongoing pregnancy rate was 39.2% per embryotransfer (ET) and 343% per started cycle. A high implantationrate was also achieved (26.6% in non-obstructive and 30% inobstructive azoospermia group). Using testicular spermatozoain combination with ICSI in both obstructive and non-obstructiveazoospermic groups, high implantation and pregnancy rates canbe achieved.     

Intracytoplasmic sperm injection in obstructive and non-obstructive azoospermia

We compared the results of intracytoplasmic sperm injection (ICSI) in: (i) obstructive versus non-obstructive azoospermia, (ii) obstructive azoospermia using epididymal versus testicular spermatozoa and (iii) acquired versus congenital obstructive azoospermia due to congenital absence of the vas deferens (CAVD). A retrospective analysis was done of 241 consecutive ICSI cycles done in 103 patients with non- obstructive azoospermia and 119 patients with obstructive azoospermia. In the obstructive group, 135 ICSI cycles were performed. Epididymal spermatozoa were used in 44 cycles and testicular spermatozoa in 91 cycles. In the non-obstructive group, 106 cycles were performed. The fertilization and pregnancy per cycle rates were 59.5 and 27.3% respectively using epididymal spermatozoa, 54.4 and 31.9% respectively using testicular spermatozoa in obstructive cases, and 39 and 11.3% respectively in non-obstructive cases. The fertilization and pregnancy per cycle rates were 56.6 and 37% respectively in acquired obstructive cases, and 55.2 and 20.4% respectively in CAVD. In conclusion, ICSI using spermatozoa from patients with acquired obstructive azoospermia resulted in significantly higher fertilization and pregnancy rates as compared to CAVD and non-obstructive cases.      

Fertilization, pregnancy and embryo implantation rates after ICSI in cases of obstructive and non-obstructive azoospermia

The aetiology of azoospermia can be grossly divided into obstructive and non-obstructive causes. Although in both cases testicular spermatozoa can be used to treat male fertility, it is not well established whether success rates following intracytoplasmic sperm injection (ICSI) are comparable. Therefore, a retrospective analysis of fertilization, pregnancy and embryo implantation rates was performed following ICSI with testicular spermatozoa in obstructive or non-obstructive azoospermia. In total, 193 ICSI cycles were carried out with freshly retrieved testicular spermatozoa; in 139 cases of obstructive and 54 cases of non-obstructive azoospermia. The fertilization rate after ICSI with testicular spermatozoa in non-obstructive azoospermia was significantly lower than in obstructive azoospermia (67.8% versus 74.5%; P = 0.0167). Within the non-obstructive group, the fertilization rate in the group of maturation arrest (47.0%) was significantly lower than in case of Sertoli cell-only (SCO) syndrome (71.2%) or germ cell hypoplasia (79. 5%). Embryo quality on day 2 after ICSI was similar for all groups. Pregnancy rates per transfer between obstructive (36.8%) and non-obstructive groups (36.7%) were similar. In cases of maturation arrest the pregnancy rate per transfer was lowest (20.0%) although not significantly different from SCO syndrome or hypoplasia groups. Embryo implantation rates were not different between the obstructive (19.6%) and non-obstructive groups (25.8%), and were lowest in cases of germ cell hypoplasia (15.8%). This retrospective analysis shows that although fertilization rate after ICSI with testicular spermatozoa in non-obstructive azoospermia is significantly lower than in obstructive azoospermia, pregnancy and embryo implantation rates are similar.     

Fertilization and pregnancy outcome with intracytoplasmic sperm injection for azoospermic men

The evident ability of the intracytoplasmic sperm injection (ICSI) procedure to achieve high fertilization and pregnancy rates regardless of semen characteristics has induced its application with spermatozoa surgically retrieved from azoospermic men. Here, ICSI outcome was analysed in 308 cases according to the cause of azoospermia; four additional cycles were with cases of necrozoospermia. All couples were genetically counselled and appropriately screened. Spermatozoa were retrieved by microsurgical epididymal aspiration or from testicular biopsies. Epididymal obstructions were considered congenital (n = 138) or acquired (n = 103), based on the aetiology. Testicular sperm cases were assessed according to the presence (n = 14) or absence (n = 53) of reproductive tract obstruction. The fertilization rate using fresh or cryopreserved epididymal spermatozoa was 72.4% of 911 eggs for acquired obstructions, and 73.1% of 1524 eggs for congenital cases; with clinical pregnancy rates of 48.5% (50/103) and 61.6% (85/138) respectively. Spermatozoa from testicular biopsies fertilized 57.0% of 533 eggs in non-obstructive cases compared to 80.5% of 118 eggs (P = 0.0001) in obstructive azoospermia. The clinical pregnancy rate was 49.1% (26/53) for non-obstructive cases and 57.1% (8/14) for testicular spermatozoa obtained in obstructive azoospermia, including three established with frozen-thawed testicular spermatozoa. In cases of obstructive azoospermia, fertilization and pregnancy rates with epididymal spermatozoa were higher than those achieved using spermatozoa obtained from the testes of men with non-obstructive azoospermia.     

The outcome of intracytoplasmic injection of fresh and cryopreserved epididymal spermatozoa from patients with obstructive azoospermia--a comparative study

The aim of our study was to compare the outcome of intracytoplasmic sperm injection (ICSI) with fresh and frozen-thawed epididymal spermatozoa retrieved by percutaneous epididymal sperm aspiration (PESA) or microepididymal sperm aspiration (MESA) from patients with obstructive azoospermia. A retrospective analysis of consecutive ICSI cycles was performed, comparing the outcome in 24 patients with obstructive azoospermia undergoing surgical sperm aspiration by MESA (7 cycles) or PESA (17 cycles). In 23 of 24 patients, excess spermatozoa were cryopreserved. Following thawing, 21 ICSI cycles were performed (11 cycles after MESA, 10 after PESA). No statistically significant differences were noted in all parameters examined in ICSI cycles with fresh or cryopreserved spermatozoa from the same patients. Comparing all ICSI cycles with fresh and frozen-thawed epididymal spermatozoa, the rates of two-pronuclear fertilization (56% versus 53%), embryo cleavage (90% versus 86%), implantation (10% versus 14%), clinical pregnancy per embryo transfer (32% versus 37%) and delivery/ongoing pregnancy rate (27% versus 26%) were not statistically different. The cumulative ongoing pregnancy rate per sperm retrieval procedure was 46%, respectively. We conclude that the clinical outcome of ICSI with fresh and frozen-thawed spermatozoa after retrieval by PESA was similar to that by MESA. Epididymal sperm cryopreservation in patients with obstructive azoospermia is feasible and efficient using a simple freezing protocol and should be offered to optimize the yield of pregnancies achieved following such procedures.      

Chromosome analysis of epididymal and testicular sperm in azoospermic patients undergoing ICSI

BACKGROUND: Although ICSI provides a way of treating azoospermic men, concern has been raised about the potential risk for transmission of genetic abnormalities to the offspring. We quantified the incidence of chromosomal abnormalities in epididymal and testicular sperm retrieved from azoospermic patients undergoing ICSI. METHODS: Individual testicular sperm were collected from testicular biopsies with an ICSI pipette, and epididymal sperm were retrieved by microsurgical epididymal sperm aspiration. Samples were processed by fluorescent in-situ hybridization (FISH) for chromosomes 18, 21, X and Y and the results compared with those from normal ejaculated samples. RESULTS: The overall aneuploidy rate of 11.4% in men with non-obstructive azoospermia was significantly higher (P = 0.0001) than the 1.8% detected in epididymal sperm from men with obstructive azoospermia and also the 1.5% found in ejaculated sperm. No significant difference was found between the epididymal and ejaculated samples. When the chromosomal abnormalities were analysed, gonosomal disomy was the most recurrent abnormality in both obstructive and non-obstructive azoospermic patients, while autosomal disomy was the most frequent in ejaculated sperm. CONCLUSIONS: Sperm of non-obstructive azoospermic men had a higher incidence of chromosomal abnormalities, of which sex chromosome aneuploidy was the most predominant. Genetic counselling should be offered to all couples considering infertility treatment by ICSI with testicular sperm.