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。     

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。     

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.     

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，     

采用经皮抽吸取精法结合卵胞浆内单精子注射治疗无精子症52例临床报道

在不孕不育夫妇中,由男性因素导致不育的夫妇占40%。自1992年Palermo等[1]首次用卵胞浆内单精子注射术（intra-cytoplasmic sperm injection,ICSI）治疗男性少,弱精子症的有效方法以来,使无精症等疾病的患者也有了生育后代的希望。近几年来,由于ICSI技术的不断发展和提高,以治疗男性不育     

不同来源精子对ICSI助孕结局的影响

目的探讨不同来源精子对卵胞浆内单精子注射助孕结局的影响。方法回顾分析我中心2008年1月至2012年6月1129个ICSI治疗周期,射出A组858例,附睾穿刺B组229例,睾丸穿刺C组42例,比较3组的临床结局。结果 A组年龄较B、C组大,内膜厚度薄;C组获卵数及ICSI卵数较A、B组多;B组优质胚胎率高于A组,种植率及活产率高于A、C两组（P〈0.05）。通过多元逻辑斯蒂回归模型分析精子来源与活产率的相关性,校正混杂因素后,A组相对于B组的OR值为0.559（95%CI 0.416-0.752）,C组相对于B组的OR值为0.513（95%CI 0.264-0.998）。结论附睾穿刺来源精子行ICSI可获得较好的临床结局。     

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。     

无精子症患者附睾穿刺及睾丸活检结果分析

无精子症占男性不育病例的10％左右,其病因多不确定,治疗效果也不好。本文通过对无精子症患者经皮附睾精子抽吸术（PESA）及睾丸活检（TESE）,分析睾丸病变,为其病因分析、临床诊疗、辅助生殖等提供实验室依据。     

两种TESA方法在腮腺炎致无精子症患者取精中的比较

目的探讨腮腺炎致无精症患者两种睾丸活检取精（TESA）即开放性睾丸活检和显微切割睾丸活检取精的优劣。方法每组24例患者,分别用开放性睾丸活检和显微切割睾丸活检取精,观察取精成功率、术后并发症发生率,术后并发症包括术后血肿的形成,术后3月睾丸萎缩、睾酮下降。结果开放性睾丸活检取精组,能找到活动的精子14例,取精成功率58．3％,术后并发症10例,约41．7％,分别为血肿2例、三月后睾丸萎缩3例、睾酮下降5例;显微切割睾丸活检取精组,能找到活动的精子18例,取精成功率75％,术后并发症5例,约20．8％,分别为血肿0例、三月后睾丸萎缩1例、睾酮下降4例。取精成功率和开放性睾丸活检取精组比较（P〈0．05）,术后并发症和开放性睾丸活检取精组此较（P〈0．01）。结论显微切割睾丸活检取精较开放手术睾丸活检取精成功率高、术后并发症少,值得推广。     

Studies of percutaneous epididymal sperm aspiration (PESA) and intracytoplasmic sperm injection

Four distinct studies were carried out using two data sets ofpercutaneous epididymal sperm aspiration (PESA) and intracytoplasmicsperm injection (ICSI) procedures performed from March 1993to January 1997. In study A, an analysis of 181 ICSI treatmentcycles following PESA revealed a successful epididymal spermretrieval rate of 83%. It confirmed that PESA is an effectivesperm retrieval method and the associated ICSI pregnancy rate(35% per embryo transfer) compared favourably with that of othersperm retrieval methods. In study B, the relevance of a priordiagnostic PESA procedure was ascertained by comparing the spermretrieval rates in two groups of patients having their firstICSI treatment cycle with spermatozoa retrieved through PESA.Group B1 (n=50) had diagnostic PESA prior to the ICSI treatmentcycle PESA procedure, unlike patients in group B2 (n=64) whodid not. The sperm retrieval rate in the treatment cycle procedurewas not different at 90 and 82.8% for groups B1 and B2 respectively.However, the discontinuation of diagnostic PESA is fraught withproblems including liability to medico-legal sanctions. In studyC, analysis of 177 treatment cycles involving PESA and ICSIrevealed a successful sperm retrieval rate by PESA of 82% inthe first cycle, 93% in the second, 96% in the third and 100%in the fourth cycle. The same trend was evident when sperm retrievalwas examined in relation to each of the epididymides. Retrievedspermatozoa were found to be motile in 67-100% of cases andthe frequency of samples containing motile spermatozoa did notdecrease with increase in the number of PESA attempts. Theseresults show that PESA does not jeopardize future epididymalsperm retrieval. In study D, the outcome of treatment with ICSIusing ejaculated spermatozoa (305 cycles) (group D1) was comparedwith that of ICSI using spermatozoa obtained through PESA (54cycles) (group D2). The median age of women in the two groupsof couples was similar (34 years). In group D1, 70% of metaphaseII oocytes were fertilized compared with 61% in group D2 (P<0.01).The cleavage rate and the median numbers of transferred andcryopreserved embryos were similar in both groups. There wasno significant difference between the clinical pregnancy rates(33 and 42% in groups D1 and D2 respectively). Our results showthat the outcome of PESA-ICSI treatment compared favourablywith that of ICSI using ejaculated spermatozoa.     

Preliminary evidence on the safety of ICSI with testicular spermatozoa in HCV-infected male: a case report

An infertile couple with a hepatitis C (HCV) RNA-positive male came to our attention. We were not able to perform an assisted reproduction technology (ART) procedure with ejaculated spermatozoa free of HCV RNA using gradient centrifugation and swim up, due to retrograde ejaculation and severe male factor. ICSI with testicular spermatozoa was the most rational therapeutic approach. The couple was informed about the lack of adequate data on the safety of this therapeutic approach. The risks of this procedure were accepted by the couple. Testicular sperm aspiration combined with ICSI (TESA-ICSI) was performed. After a negative result for an HCV RNA PCR on the embryos' culture medium, four embryos were transferred to the uterine cavity on the third day. The procedure resulted in an ongoing clinical pregnancy, and HCV antibody determinations performed in the mother at the 12th and 24th week of pregnancy were negative. The pregnancy ended at the 39th week due to endo-uterine death. No malformation or hepatic pathologies were found in the conceptus. A second TESA-ICSI cycle is ongoing. This preliminary evidence suggests that, in HCV sero-positive males, ICSI with testicular spermatozoa may be a safe procedure. However, we need more observations to clarify if ART is really able to reduce horizontal and vertical transmission of HCV in sero-discordant couples (only the male infected) in comparison with natural conception.     

Decreased fertilization rate and embryo quality after ICSI in oligozoospermic men with microdeletions in the azoospermia factor c region of the Y chromosome

Microdeletions of the azoospermia factor (AZF) region of the Y chromosome occur in between 1 and 29% of oligozoospermic and azoospermic men, and most deletions are found in the AZFc region. These men can father children when intracytoplasmic sperm injection (ICSI) is used, but the success rate is unclear. Thus, the success rate of 19 ICSI treatments in eight couples with a microdeletion in the AZFc region of the Y chromosome was analysed retrospectively. These were compared with a control group of 239 ICSI treatments in 107 couples undergoing ICSI treatment with ejaculated spermatozoa. The fertilization rate was significantly lower in the group of Y-deleted men (55%; 95% CI: 41-69%) compared with controls (71%; 95% CI: 67-74%; P < 0.01). The embryo quality was also significantly poorer among Y-deleted men (P<0.001). Pregnancy, implantation and take-home baby rates were not significantly lower in the Y-deleted group. This study shows that ICSI in oligozoospermic men with microdeletions in the AZFc region of the Y chromosome leads to a lower fertilization rate and poorer embryo quality.     

Testicular sperm extraction (TESE) and ICSI in patients with permanent azoospermia after chemotherapy

BACKGROUND: Patients persistently azoospermic after chemotherapy have been considered traditionally as sterile unless sperm was frozen before therapy. Recent advances during the last decade combining testicular sperm extraction (TESE) and ICSI in patients with non-obstructive azoospermia allow these males to father their own genetic offspring. METHODS: A retrospective study was conducted of 12 patients with non-obstructive azoospermia after chemotherapy undergoing TESE between 1995 and 2002. Cancer type and anti-neoplastic treatments were recorded, together with maximum testicular volume, serum FSH levels and testicular histopathology. When TESE was successful, spermatozoa were cryopreserved for performing ICSI later. RESULTS: In five patients (41.6%) motile spermatozoa for cryopreservation and ICSI were retrieved. Four of them had received chemotherapy for testicular cancer, and one had been treated by chemotherapy/radiotherapy for Hodgkin's disease. Clinical and histological parameters were unable to predict with certainty TESE outcome in an individual patient. Eight ICSI cycles were performed on five couples and one pregnancy was obtained which resulted in the delivery of a healthy girl. CONCLUSION: Some patients with permanent azoospermia after chemotherapy can be successfully treated by TESE-ICSI. This procedure, however, may have potential genetic risks. Therefore, freezing semen before starting gonadotoxic therapy is the strategy of choice, and patients should be counselled accordingly.     

ICSI outcome in patients with transient azoospermia with initially motile or immotile sperm in the ejaculate

BACKGROUND: In patients with transient azoospermia, few sperm may be found in the ejaculate. We investigated the outcome of ICSI in patients with transient azoospermia. METHODS: Records of patients with transient azoospermia referred during a 42 month period were reviewed. If only immotile sperm were found, the sample was incubated with 30% human serum albumin (HSA) before motility re-assessment. If still immotile, mechanical assessment of sperm viability was utilized. Study groups were: (A) motile sperm; (B) motility achieved by HSA; (C) no motility, but viability assessed by a mechanical technique; and (D) control group with sperm counts from 1 to 5 x 10(6)/ml. There were 57 couples (cycles) in the study group and 43 couples (cycles) in the control group. RESULTS: Age, days of stimulation and endometrial thickness were comparable among groups. In 29.8% of the cycles, only immotile sperm were found. Fertilization and cleavage rates were higher in groups A and D than in groups B and C. Clinical pregnancy rate/cycle and live birth rate/cycle were not different among groups. No congenital malformations were found in newborns. CONCLUSION: Fertilization and cleavage rates were lower in patients with initially immotile sperm compared with those with initially motile sperm and oligoasthenoteratozoospermia patients. Clinical pregnancy and viable pregnancy rates were not statistically different among groups, although when only immotile sperm were present both clinical pregnancy and live birth rate were lower in comparison with cycles with motile sperm.     

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.     

男性不育的遗传学研究及在ICSI中的意义

目的在细胞遗传学基础上,建立和应用一套检测Y染色体微缺失和雄激素受体基因外显子A突变的分子诊断方法,以便研究男性不育发病机制,为临床辅助生殖技术提供遗传咨询。方法对139例原发不育患者采用外周血染色体G显带、C显带技术,并选择其中40例应用多重PCR和PCR-SSCP银染进行DAZ基因及雄激素受体基因外显子A突变检测。结果139例原发不育患者G、C显带发现54例染色体核型异常,3例DAZ基因缺失,5例雄激素受体基因外显子A发生点突变。结论通过细胞遗传学检查和DAZ基因及雄激素受体基因外显子A突变检测,从遗传学角度探讨了男性不育的病因,对卵浆单精子显微注射技术提供了理论依据。     

Achievement of pregnancy in globozoospermia with Y chromosome microdeletion after ICSI

Pregnancy achieved with sperm from a patient with globozoospermia is rare, even after ICSI, since the activation of the oocyte may not occur in this disorder. Therefore, activation of the oocytes by piezoelectricity or calcium ionophores has been suggested, although spontaneous activation of the oocyte after ICSI has been reported in some cases. We report a successful pregnancy in a couple in which the male partner had globozoospermia with microdeletions in the Y chromosome with no further assisted activation after ICSI. During the diagnostic study of the husband, increased numerical chromosome abnormalities after fluorescent in-situ hybridization (FISH) and microdeletions in AZFa; sY86 and AZFb; sY 131 were detected. Out of the 13 oocytes injected, four fertilized and a twin pregnancy was obtained after replacement of four embryos. Healthy twin girls were delivered after a term pregnancy. Some patients with globozoospermia may also have Y chromosome microdeletions, which subsequently may be inherited by the male offspring in cases of achievement of pregnancy.