采用抽吸取精法结合单精子注射治疗无精子症结局

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（testicular 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子注射。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73.0%,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。     

显微取精术在非梗阻性无精子症患者中的临床应用及结局

目的观察睾丸显微取精术在非梗阻性无精子症患者中的临床应用及其结局。方法17例患者在配偶同步促排卵取卵前一天行睾丸显微切开取精术,获得精子患者行卵胞浆内单精子显微注射(ICSI)助孕,分析其受精率、可用胚胎率和临床妊娠结局。结果17例患者显微取精,13例成功获取精子,精子获得率为76.5%。这13对夫妇ICSI受精率为74.8%,可用胚胎率49.5%。3例新鲜周期移植,全部成功妊娠,1例已分娩1健康婴儿,另外2例持续妊娠中。其余10例行全胚冷冻,4例分别于胚胎冻存3个月后行解冻复苏移植,3例成功妊娠,2例已分娩(其中一例为非嵌合型Klinefelter综合征患者),另外1例持续妊娠中。结论睾丸显微切开取精术是使NOA患者和非嵌合型Klinefelter综合征患者成功获得自己遗传学子代的有效方法,有较高的精子获得率,联合ICSI技术、全胚冷冻-复苏移植可以获得较高的临床妊娠率。     

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。     

细胞周期类分子在人无精子症睾丸中的表达

目的:评价细胞周期类基因在人无精子症及正常睾丸组织中的表达及意义.方法:应用包含有人CDC10等细胞周期类基因在内的cDNA微矩阵芯片对人正常睾丸及无精子症睾丸组织中差异表达基因进行了研究:通过PCR方法获得两种组织mRNA, 再分别用Cy5-dUTP及Cy3-dUTP标记制备cDNA探针.两种探针混合后与人cDNA微矩阵芯片杂交, 经扫描、计算机处理分析比较杂交结果;利用原位杂交技术对芯片杂交结果进行了验证研究.结果:部分细胞周期类基因可能与无精子症相关, 其中CDC7L1 与CDC10基因表达上调, CDK9、 CDC20 以及CLK3基因表达下调.原位杂交证实CDC10在正常睾丸组织生精细胞中表达强于无精子症睾丸组织.结论:细胞周期类分子CDC10、 CDC7L1 、 CDK9、 CDC20及CLK3可能在无精子症的发生与进展过程中起一定的作用.     

无精子症患者睾丸生精功能与血清性激素水平的相关性分析

目的通过对广东佛山地区104例无精子症患者的血清性激素水平进行检测并分析,探讨其与睾丸生精功能的相关性。方法 104例研究对象均于我院确诊为无精子症患者,测定其血清中总睾酮(T,nmol/L)、卵泡刺激素(FSH,IU/L)、黄体生成素(LH,IU/L)、泌乳素(PRL,ng/m L)、雌二醇(E2,ng/L)水平,根据睾丸活检病理结果分为生精功能正常组(1组)、生精功能低下组(2组)、唯支持细胞综合征组(3组)。结果各组患者的年龄均无统计学差异,1组与2组患者各性激素水平无统计学差异;3组患者血清FSH、LH、PRL水平高于2组,E2水平低于2组,有显著的统计学差异,虽然3组T水平低于2组,但两组间没有统计学差异;3组患者血清FSH、LH水平高于1组,T水平低于1组,有显著的统计学差异,但两组之间的PRL、E2水平并无统计学差异。相关分析显示睾丸生精功能与T水平呈正相关,与FSH、LH水平呈负相关,与PRL、E2水平没有相关性。结论血清性激素水平测定对于预测无精子症患者睾丸生精功能有重要意义,并可用于指导治疗及判断预后情况。     

无精子症患者血清中FSH的测定及临床意义

目的通过检测无精子症患者血清FSH水平,分析鉴别梗阻性无精子症和非梗阻性无精子症两种类型;通过比较血清FSH水平与睾丸体积,分析血清FSH水平与睾丸体积的关系,探讨其判断无精子症分型的意义。方法对2007年10月-2008年4月间76例来自大连市妇产医院白山路生殖中心的无精子症患者的血液样本,采用ROCHE2010全自动免疫分析仪及其配套试剂盒检测血清FSH水平,采用Prader睾丸模型比拟法测定睾丸体积。结果与对照组相比,非梗阻性无精子症组血清FSH水平显著升高（P〈0.05）,而对照组与梗阻性无精子症组血清FSH水平差异无显著性（P〉0.05）。非梗阻性无精子症组与梗阻性无精子症组血清FSH水平差异显著（P〈0.05）。与睾丸体积≥12ml组相比,睾丸体积〈12ml组血清FSH水平显著升高（P〈0.05）。以睾丸体积12ml为分界标准,≥12ml为梗阻性无精子症,〈12ml为非梗阻性无精子症。该标准对梗阻性和非梗阻性无精子症诊断的符合率分别为89.3%（25/28）、75%（36/48）。以生殖激素中FSH12.4mIU/ml为分界标准,小于该值为梗阻性无精子症,大于该值为非梗阻性无精子症,该标准对梗阻性无精子症诊断的符合率为92.9%（26/28）,对非梗阻性无精子症诊断的符合率为89.6%（43/48）。结论血清FSH水平为分析睾丸功能衰竭原因提供依据,对判定精道梗阻具有重要意义,是鉴别非梗阻性与梗阻性无精子症的首要临床指标;睾丸体积在判定无精子症分型中有一定意义;血清FSH水平与睾丸体积联合检测更有助于鉴别梗阻性无精子症和非梗阻性无精子症。     

睾丸精子抽吸后行卵细胞单精子显微注射治疗梗阻性无精子症

无精子症约占男性不育患者的7％～14％。目前尚无确切的治疗方法。自Palerm等于1992年首次报道卵细胞浆内单精子注射（ICSI）获得妊娠成功，为男性不育症治疗提供了新方法，同时附睾或睾丸取精术彻底改变了无精子症不可治疗的局面。采用此方法只要在男性生殖道或睾丸内发现并分离到精子，利用ICSI技术就能获得一定的妊娠率，     

少弱精子症、死精子症、无精子症患者细胞遗传学分析

目的了解少弱精子症、死精子症、无精子症与染色体之间的联系。方法对2011年上半年生殖门诊101例少弱精子症、死精子症、无精子症患者,进行染色体检测,并对结果进行比较分析。结果染色体异常总数占23例,异常发生率达22.8%。性染色体异常9例,占异常总数的39%,常染色体异常14例,占异常总数61%。结论男性不育少弱精子症、死精子症、无精子症患者常规做染色体检查是必要的,对确定其是否有治疗价值提供重要依据。     

无精子症和少精子症患者细胞遗传学分析

男性不育是生育异常中较常见的疾病 ,也是人类生殖研究中一项重要课题。男性不育通常与无精子症和少精子症有着密切的关系 ,染色体异常是其中重要原因之一。我室自1987年至今分别对 6 8例无精子症和 82例少精子症患者进行了外周血染色体检查。现将结果报道分析如下。资料与方法     

Outcome of testicular sperm retrieval procedures in non-obstructive azoospermia: percutaneous aspiration versus open biopsy

The aim of this study was to evaluate whether the extraction of testicular spermatozoa with percutaneous versus open biopsy has an effect on the treatment outcome with intracytoplasmic sperm injection (ICSI) in men with non-obstructive azoospermia. Regardless of testicular size, follicle stimulating hormone concentration, and previous biopsy result, percutaneous testicular sperm aspiration (PTSA) using a 21-gauge butterfly needle was attempted first and if this failed testicular sperm extraction (TESE) was performed. In 63 men spermatozoa were found with PTSA whereas in 228 men TESE had to be undertaken. More men in the PTSA group had previously been diagnosed with hypospermatogenesis (82 versus 50%). Compared with the PTSA group, more men in the TESE group had germ cell aplasia (27 versus 10%) or maturation arrest (22 versus 8%). There was no difference between the groups regarding mean age of men and their partners, duration of stimulation, oestradiol concentration on the day of human chorionic gonadotrophin, number of oocytes retrieved, fertilization rate, and embryo quality between the two groups. The number of embryos transferred (4.38 versus 3.90) was significantly higher in the PTSA group (P < 0.05), reflecting the increased number of embryos available for transfer. Implantation rate per embryo was 20.7% in the PTSA and 13.3% in the TESE group (P < 0.05). Clinical pregnancy rates were 46 and 29% in the PTSA and TESE groups respectively (P < 0.05). Clinical abortion rates were similar (21.2 versus 24%). It is concluded that in men with non-obstructive azoospermia, easier sperm retrieval, which is most likely indicative of a more favourable histopathology, is associated with higher implantation rates per embryo.     

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.     

Testicular sperm retrieval and cryopreservation prior to initiating ovarian stimulation as the first line approach in patients with non-obstructive azoospermia.

The potency for fertilization and successful implantation was compared between fresh and cryopreserved testicular spermatozoa obtained from the same patient with non-obstructive azoospermia. Spermatozoa cryopreserved at the outset were also evaluated. Non-obstructive azoospermic men (n = 55) underwent testicular sperm extraction (TESE); mature spermatozoa were found in 33 (60%) of them. Of 57 intracytoplasmic sperm injection (ICSI) cycles in 25 patients, 15 used fresh spermatozoa (14 patients, group 1), 24 used the excess spermatozoa cryopreserved after 'fresh' ICSI (11 couples who did not conceive in the 'fresh' cycle, group 2) and 18 cycles used cryopreserved spermatozoa at the outset (11 other patients, group 3). Fertilization, cleavage, embryo quality, implantation and take home baby rates were not significantly different in groups 1 and 2, and 6/14 couples ultimately had healthy babies (42.8% cumulative take home baby rate per TESE). In group 3, neither the fertilization rate, embryo development, pregnancy nor implantation rates per embryo transfer were significantly different from groups 1 and 2. The cumulative delivery and ongoing pregnancy rate in this group was 36. 4%. Cryopreservation did not impair the availability of motile spermatozoa for ICSI. When immotile spermatozoa were injected, however, fertilization rate decreased dramatically. Since criteria for predicting the presence of spermatozoa in the testicular tissue of patients with non-obstructive azoospermia are inadequate, it is suggested that TESE be performed prior to initiating ovarian stimulation.     

Outcome of intracytoplasmic sperm injection with testicular spermatozoa in obstructive and non-obstructive azoospermia

From 1 August 1993 until 30 September 1994, 69 couples sufferingfrom azoospermia underwent testicular sperm extraction and intracytoplasmicsperm injection. In 50 couples with obstructive azoospermiaa total of 631 meta-phase-II oocytes were injected after testicularsperm extraction yielding a 2-PN fertilization rate of 57%.In female patients <40 years of age an ongoing pregnancyrate per transfer of 42% (14/33) was obtained. So far, eighthealthy babies have been born, including two singletons andthree twin gestations. In 19 couples with non-obstructive azoospermiaa total of 264 metaphase-II oocytes were injected after testicularsperm extraction, yielding a 2-PN fertilization rate of 58%.An ongoing pregnancy rate per transfer of 31% (5/16) was established.So far, six healthy babies have been born including one singleton,one twin and one triplet gestation.     

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.     

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.     

Fertility with testicular sperm extraction and intracytoplasmic sperm injection in non-obstructive azoospermic men

In non-obstructive azoospermia spermatozoa can usually onlybe isolated from the testicles, and thus the most promisingtreatment model is testicular sperm extraction (TESE). Hormoneconcentrations, testicular volume determinations and testicularbiopsy results are not uniform enough to select potential candidatesfor successful TESE and intracytoplasmic sperm injection (ICSI)approaches in advance. The aim of this study was to assess theefficacy of using ICSI with testicular spermatozoa in casesof non-obstructive azoospermia and to compare the inclusioncriteria and sperm existence in the testicles in sperm obtainableand non-obtainable groups. All men showed either complete orincomplete (n = 14) maturation arrest in spermatogenesis, severehypospermatogenesis (n = 10) or Sertoli cell-only syndrome (n= 5) in their testicular biopsies. Only 14 out of a total of29 men provided enough spermatozoa for the ICSI procedure, whileno spermatozoa were found in the testicular samples of the remaining15 men. Out of 123 oocytes obtained from 14 females, 101 wereinjected with the husbands' testicular sperm cells. Total fertilizationfailure was observed in three cases. Of 39 oocytes fertilized,38 cleaved. The fertilization and cleavage rates were 38.6 and97.4% respectively. The pregnancy rate was 20.7% per initiatedcycle. In the group from whom spermatozoa were obtainable, thepregnancy rate was 42.9% per initiated cycle and 54.5% per embryotransfer. A total of six pregnancies were achieved, of whichtwo Were twins and four were singletons. One singleton pregnancyresulted in abortion in the first trimester. There was no statisticaldifference concerning the serum follicle stimulating hormoneconcentration, testicular volume and biopsy results in groupsin which spermatozoa were obtainable or not. In conclusion,although the association of TESE with ICSI obtained pregnanciesfor some patients with non-obstructive azoospermia, furtherstudies are needed to determine the inclusion criteria for successfulTESE.     

Outcome of first and repeated testicular sperm extraction and ICSI in patients with non-obstructive azoospermia

BACKGROUND: It is unclear whether or not testicular sperm extraction (TESE) should be repeated for patients in whom no sperm were found during their first TESE attempt. METHODS AND RESULTS: The outcome of repeated TESE was evaluated in patients with non-obstructive azoospermia (NOA) after failing to obtain sperm in their first extraction attempt, or having used all available cryopreserved testicular tissue. Out of 83 patients with NOA, patients repeated TESE two (n = 22), three (n = 8), four (n = 6) and five (n = 3) times. Distribution of main testicular histology included germ cell aplasia (55%), maturation arrest (29%) and germ cell hypoplasia (16%). The first TESE yielded mature sperm for ICSI in 39% of patients (sp+), and failed in the remaining 61% (sp-). A second TESE yielded mature sperm in 1/4 from the sp- group and in 16/18 from the sp+ group. At the third, fourth and fifth trials, 8/8, 5/6 and 3/3 of the original sp+ patients were sp+ again respectively. Compared with the outcome of the first trial, all further trials did not differ statistically in the rate of fertilization (54 versus 49%), implantation (9.5 versus 5.4%), or clinical pregnancy/cycle (19 versus 15%). No pregnancies were achieved among the three patients after their fifth TESE. Pregnancies occurred in all histological groups, except maturation arrest. CONCLUSIONS: The outcome of repeated TESE cycles, up to the fourth trial, justifies the procedure.