附睾和睾丸精子活力对卵胞浆内单精子注射结局的影响

目的比较附睾或睾丸来源及其不同活力精子行卵胞浆内单精子注射（ICSI）的结局。方法回顾性分析2005年1月至2008年5月在本生殖中心经皮附睾精子抽吸术（PESA）、睾丸精子抽吸术（TESA）助孕的218例无精子症患者的资料,比较附睾和睾丸及其不同活力精子的正常受精率、卵裂率、优质胚胎率、临床妊娠率、种植率和早期流产率。结果附睾精子组与睾丸精子组比较,正常受精率、卵裂率、临床妊娠率、种植率和早期流产率无显著性差异（P〉0．05）;优质胚胎率附睾精子组显著高于睾丸精子组（P〈0．05）。附睾活动精子、睾丸活动精子和睾丸不活动精子的正常受精率显著高于附睾不活动精子（P〈0．01）;睾丸活动精子的正常受精率显著高于睾丸不活动精子组（P〈0．05）;以上各组间的优质胚胎率、临床妊娠率、种植率、流产率均无显著性差异（P〉0．05）。结论PESA操作简单且不影响妊娠率,无精子症患者行ICSI治疗时可首选附睾精子;附睾或睾丸不活动精子影响ICSI的受精率,应优先选择活力较好的精子,若无活动精子则选择睾丸不活动精子。     

不同生精功能和取精方式对无精子症ICSI妊娠结局的影响

目的 分析采用经皮附睾精子抽吸术(PESA)或经皮睾丸精子抽吸术(TESA)获得精子对不同生精功能无精子症进行卵泡浆内单精子注射(ICSI)治疗的妊娠结局.方法 经PESA获得附睾精子,经TESA获得睾丸精子,女方进行常规超排卵.两种取精方法获得的精子进行ICSI,比较其妊娠率.结果 216次采用PESA获得附睾精子,87次采用TESA获得睾丸精子,PESA和TESA组的妊娠率分别为41.7%和43.7%,(P>0.05).随着生精功能状态从正常到重度生精功能障碍的变化,其妊娠率依次为:46.8%,41.6%,36.7%和16.7%.其中生精功能正常组与轻度和中度生精功能障碍组差异无统计学意义,但与重度生精功能障碍组差异均有统计学意义.结论 采用PESA或TESA结合ICSI是治疗男性无精子症的有效方法,而且认为生精功能正常组和轻度及中度生精功能障碍三组无精子症患者均可试行ICSI.     

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 )。     

微创取精术结合ICSI治疗阻塞性无精子症的研究

目的探讨采用微创手术从阻塞性无精子症患者获取精子,再行卵母细胞单精子显微注射(ICSI)及胚胎移植治疗不育症的效果。方法对20例阻塞性无精子症患者先行经皮附睾精子抽吸术(PESA),如果未取到精子,再行睾丸精子穿刺吸取术(TESA),随后行ICSI辅助生育治疗周期共20次。结果20例病人中,采用PESA成功获得精子者共18例,未成功者2例,进一步行TESA后,均获得精子。用取得的精子行MⅡ期卵子ICSI后,受精率达68．09%,在20例胚胎移植后获得临床妊娠9例(45%)。结论对于阻塞性无精子症患者,应用微创取精术可以成功获得精子,行ICSI治疗后可获得满意的受精率和临床妊娠率,微创取精术可以成为阻塞性无精子症患者获得精子的首选方法。     

附睾及睾丸精子行ICSI治疗无精子症妊娠结局

目的 :回顾性分析 5 0例无精子症患者利用附睾或睾丸精子行卵细胞胞质内单精子注射 (ICSI)的治疗结局。　方法 :经皮附睾精子抽吸术 (PESA)或睾丸切开取精术 (TESE)获得精子行ICSI,评估取精的成功率 ,ICSI后的受精率、种植率及临床妊娠率 ,以精液精子ICSI组作为对照。　结果 :PESA、TESE与精液精子组分别注射MⅡ期成熟卵子 2 86、36 0、15 6 9个 ,受精率 3组差异无显著性 (74 .8% ,75 .2 %vs 77.5 % ,P >0 .0 5 )。种植率、妊娠率TESE与精液精子组差异无显著性 (2 9.87%vs 2 9.5 4 % ;4 8.15 %vs 5 2 .6 0 % ,P >0 .0 5 ) ,PESA组显著高于TESE组及精液精子组 (5 0 .85 %vs 2 9.87% ,2 9.5 4 % ;6 8%vs 4 8.15 % ,5 2 .6 0 % ,P <0 .0 5 )。PESA组共妊娠 17例 ,已分娩 6例 ,继续妊娠 9例 ,流产 2例 ;TESE组共妊娠 13例 ,已分娩 7例 ,继续妊娠 4例 ,流产 2例。　结论 :采用附睾或睾丸精子行ICSI是治疗男性无精子症的有效方法。     

冻融复苏微量精子行卵细胞胞质内单精子注射术的疗效及临床妊娠结局分析

目的:回顾性分析123例无精子症患者经皮附睾精子抽吸术(PESA)或经皮睾丸精子抽吸术(TESA)后冻融复苏微量精子行卵细胞胞质内单精子注射术(ICSI)的疗效及临床妊娠结局情况。方法:将采用微量冻融PESA、TESA精子行ICSI的病例归为冻融精子组,采用新鲜PESA、TESA精子行ICSI的病例归为对照组。比较冻融精子组与新鲜精子组组间及组内的双原核(2PN)受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率、多胎妊娠率有无统计学差异。结果:PESA精子冻融组与新鲜组受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率及多胎妊娠率分别为75.67%vs76.49%,64.96%vs66.19%,55.21%vs57.22%,13.21%vs12.61%,3.77%vs5.41%,37.74%vs37.84%(P>0.05),TESA精子冻融组与新鲜组受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率及多胎妊娠率分别为74.41%vs76.43%,64.63%vs66.35%,46.81%vs53.39%,18.18%vs14.55%,4.55%vs1.82%,37.74%vs37.84%,组间及组内均无统计学差异(P>0.05)。PESA精子与TESA精子冻融复苏成功率为70.07%vs62.67%,无统计学差异(P>0.05)。结论:微量PESA及TESA精子冻融技术对无精子症患者来说是一种安全、经济、有效的治疗方法;精子冷冻复苏技术有待于进一步提高;该技术是否会增加子代远期遗传风险仍有待于进一步探讨和研究。     

外科取精术在无精子症诊断与治疗中的应用

目的研究外科取精术在无精子症诊断与治疗中的应用价值。方法在诊断为无精子症的、患者中,经睾丸体积测定、血清性激素水平、生殖系统超声等检查后,选择符合条件者198例,在局麻下行外科取精术,对获得组织显微镜下检查,统计分析取精结果。获得的精子行卵胞浆内单精子显微注射术（ICSI）及胚胎移植术（ET）,统计评估受精率、卵裂率、临床妊娠率及流产率。结果其中78例附睾中存在精子（39．4％）,23例睾丸中存在精子（11．6％）。睾丸体积正常的取精成功率明显高于睾丸体积偏小者,有显著性差异（P〈0．01）。血清促卵泡刺激素（FSH）水平正常的取精成功率明显高于FSH增高者,差异有显著性意义（P〈0．01）。82例外科取精术获得精子的患者进行ICSI治疗,附睾取精组与睾丸取精组比较,受精率、卵裂率、临床妊娠率及流产率差异均无统计学意义（P均〉0．05）。结论外科取精术操作简单且创伤较小,能准确鉴别诊断梗阻性无精子症（OA）及非梗阻性无精子症（NOA）,对无精子症的诊断有重要价值;为部分无精子症患者提供了生育自己生物学子代的机会,也是针对无精子症的有效治疗手段。     

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。     

经皮附睾精子抽吸术和睾丸精子获取术在无精子症诊断和治疗中的应用

目的 :研究附睾和睾丸精子抽吸术对无精子症患者的诊断和治疗价值。　方法 :应用经皮附睾精子抽吸术(PESA)和睾丸精子获取术 (TESE)两种方法对 385例无精子症患者进行穿刺检查。　结果 :其中 6 4例附睾中存在精子 (1 6 .6 2 %) ;4 5例患者睾丸中存在精子 (1 1 .6 9%) ;对其中 6 4例睾丸或附睾中发现精子的患者采取PESA或TESE取精后行卵细胞胞质内单精子注射 (ICSI)治疗。胚胎移植后妊娠率为 39.0 7%。　结论 :PESA和TESE为部分无精子症患者提供了生育的机会 ,也是针对无精子症的有效的治疗手段。     

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。     

Characterisation of a subpopulation of sperm with massive nuclear damage,as recognised with the sperm chromatin dispersion test

Assessment of human sperm DNA fragmentation by the sperm chromatin dispersion (SCD) test is based on the detection of haloes of spreading DNA loops after sequential DNA denaturing and protamine removal. After the SCD test, sperm without DNA fragmentation show chromatin haloes emerging from the central nuclear core, while sperm containing fragmented DNA present small or no haloes. The nuclear degraded sperm are recognised as a differentiated category within the sperm with fragmented DNA, whose cores appear irregularly and/or faintly stained. This subpopulation is more prevalent in patients with varicocele. Protein staining with 2.7‐dibrom‐4‐hydroxy‐mercury‐fluorescein demonstrated that degraded sperm intensely lose nuclear core proteins after the SCD processing. Moreover, degraded sperm are 65% more faintly labelled for DNA breaks after in situ nick translation (ISNT) on average, due to extensive DNA loss. A two‐dimensional comet assay under sequential neutral and alkaline conditions demonstrated that degraded sperm contain both massive double‐ and single‐strand DNA breaks. The degraded sperm appear as a subpopulation with stronger nuclear damage, affecting both DNA and protein fractions, possibly due to intense intratesticular oxidative stress, what could explain its higher proportion in patients with varicocele.     

Sperm selection based on motility in polyvinylpyrrolidone is associated with successful pregnancy and embryo development

The aim of this study was to investigate whether spermatozoon motility in polyvinylpyrrolidone (PVP) is associated with better embryo development and pregnancy rates in ICSI cycles. A total of 123 primary ICSI treatment cycles were included in this study. Semen samples were tested for motility before ICSI procedure in PVP. Within 3 min, the presence or absence of motility was recorded. Sperm functions were examined by the aniline blue (AB) chromatin condensation test and the hypoosmotic swelling test, and the chromatin stability was evaluated by inducing its decondensation with sodium dodecyl sulphate and ethylenediaminetetraacetic acid (EDTA). Fertilisation and embryo scoring were evaluated. Fifty (64%) of 78 women conceived in the PVP (+) group; and 12 (26%) of 45 women conceived in the PVP (?) group; the pregnancy rate was significantly higher in the PVP (+) group (P = 0.003). Semen parameters were observed to be similar in both groups. The mean number of total embryos obtained in ICSI procedure and transferred grade 1 embryos were significantly higher in PVP (+) group (P = 0.01 and P = 0.003 respectively). The presence of sperm motility in PVP is associated with increased pregnancy rate, higher percentage of good quality embryos, sperm chromatin condensation and decondensation.     

精子体外处理技术对供精人工授精妊娠结局的影响

目的探讨不同的精子体外处理技术对供精人工授精（AID）妊娠率的影响。方法对AID过程中的172个周期采用冷冻精液复苏后行宫颈人工授精（ICI），另251个周期采用复苏精液用Tyrode’s、BWW、IVF30、HTF等4种培养液经一步离心法处理后行宫腔内人工授精（IUI）和输卵管内精液灌注（FSP）。观察不同的精液处理结合不同的手术方法对AID妊娠率的影响。结果经复苏后的冷冻精液用IVF30和HTF培养后的精子的活动率较复苏精液和用Tyrode’液、BWW组显著提高（P〈0．05），而精子密度无显著差别（P〉0．05）。在431个周期中，周期妊娠率为10．90％（47／431）。复苏精液后行ICI、Tyrode’液组、BWW组、IVF30组和HTF组的周期妊娠率分别为6．98％（12／172），8．16％（8／98），9．72％（9／72），22．73％（10／44），22．22％（10／45）。5组相比较，精液经IVF30和HTF处理后行IUI或FSP的妊娠率最高（P〈0．05）。结论在进行AID时，选择不同手术方式时还必须与相应的精液处理方法相结合才能提高供精人工授精的妊娠率。     

Enhanced conception by stored porcine sperm stimulated with chloroquine

The effect of chloroquine stimulation on the motility of porcine sperm stored for 6 h and on the subsequent conception rate was assessed. There was a drastic decline in sperm motility after 6 h storage at 4 degrees and 25 degrees C, and a concomitant depression in their conception rate. However, chloroquine stimulation restored these functions towards normal, especially in sperm stored at 25 degrees C. These observations, coupled with the stability of the litter size, regardless of treatment, suggest that chloroquine can be used in an artificial insemination programme involving the utilization of semen several hours after collection and some distance away from the source of the ejaculate.     

Outcome of repeated micro-surgical testicular sperm extraction in patients with non-obstructive azoospermia

Aim： To evaluate the outcome of repetitive micro-surgical testicular sperm extraction （mTESE） attempts in non-obstructive azoospermia （NOA） cases, in relation to patients＇ initial testicular histology results. Methods： A total of 68 patients with NOA in whom mTESE had been performed in previous intracytoplasmic sperm injection （ICSI） attempts were reviewed. Results： Among the 68 patients with NOA, the first mTESE yielded mature sperm for ICSI in 44 （64%） （Sp^＋）, and failed in the remaining 24 （36%） （Sp^-）. Following their first trial, 24 patients decided to undergo a second mTESE. Of these 24 patients, no spermatozoa were obtained in 5 patients, and Sp^＋ but no fertilization/pregnancy were achieved in 19. In these 24 cases, mTESE was successively repeated for two （n = 24）, three （n = 4） and four （n = 1） times. The second attempt yielded mature sperm in 3/5 patients from the Sp group and 16/19 patients from the Sp^＋ group. At the third and fourth trials, 4/4 and 1/1 of the original Sp^＋ patients were Sp^＋ again, respectively. Distribution of main testicular histology included Sertoli cell-only syndrome （16%）, maturation arrest （22%）, hypospermatogenesis （21%） and focal spermatogenesis （41%）. Overall, in repetitive mTESE, 24/29 （82%） of the attempts were finally Sp^＋. Conclusion： Repeated mTESE in patients with NOA is a feasible option, yielding considerably high sperm recovery rate. In patients with NOA, mTESE may safely be repeated one or more times to increase sperm retrieval rate, as well as to increase the chance of retrieving fresh spermatozoa to enable ICSI.     

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.     

Parameters associated with sperm quality prior to chemotherapy in lymphoma patients

Sperm quality in lymphoma patients may be reduced even prior to initiation of chemotherapy. The objective of this study was to examine the relationship between lymphoma prognostic factors and sperm quality prior to chemotherapy. A retrospective cohort study was conducted in a Hadassah Medical Center sperm bank and the Hematology department. The cohort included 101 Hodgkin's and 90 non-Hodgkin's lymphoma patients that underwent sperm cryopreservation before chemotherapy between 1998 and 2015. Known lymphoma prognostic factors were compared between patients with normal and impaired sperm parameters. The Prognostic Score Ratio (PSR), an index representing the number of negative lymphoma prognostic measures that found in a lymphoma patient, was additionally calculated and compared between the groups. Among the prognostic factors of lymphoma, the following factors were found to be associated with impaired sperm parameters—low albumin (p < 0.001) and haemoglobin (p < 0.001) levels, B symptoms (p = 0.021) and PSR (p < 0.001). Logistic regression showed significant association of albumin and haemoglobin with reduced sperm quality (OR = 2.7 and OR = 13.5, p < 0.05; respectively). To conclude, low albumin and haemoglobin levels are related to reduced sperm quality. The linkage between these prognostic factors and sperm quality may be related to a general inflammatory status.     

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.     

Sperm ubiquitination and DNA fragmentation in men with occupational exposure and varicocele

Assessment of sperm ubiquitination and DNA fragmentation as sperm functional markers are proposed to complement routine semen analysis. This study focuses on the evaluation of these markers in infertile men with varicocele or exposed to occupational background. The results were compared with normozoospermic men. Semen parameters in both groups were lower than those in the control group. Ubiquitination median, as a marker for functionality of the ubiquitin–proteasome system, was also lower in both groups. The ubiquitination median showed a significant positive correlation with motility in both groups, while it showed only a negative correlation with sperm morphology in the varicocele group. DNA fragmentation showed a significant correlation with semen parameters, in total varicocele and also total exposure groups. In conclusion, significant difference of sperm ubiquitination between normal and study groups further validates that sperm ubiquitination as a potential molecular marker for sperm evaluation in addition to routine semen analysis in clinical laboratories.     

经皮附睾穿刺取精精子冷冻复苏后卵细胞胞质内单精子注射治疗无精子症的初步研究

目的:回顾性分析27例无精子症患者经皮附睾穿刺取精术(PESA)所获精子冷冻复苏后行卵细胞胞质内单精子注射(ICSI)治疗后的效果及妊娠结局。方法:将诊断性附睾穿刺以及PESA治疗周期ICSI后所剩余活精子以常规方法加以冷冻,将复苏后找到了足量活精子并行ICSI的病例归为冻精组,而采用新鲜PESA活精子ICSI的病例则归为对照组。比较冻精组与对照组的受精率、种植率、临床妊娠率,同时分析两组间的妊娠并发症、新生儿出生及畸形等情况。结果:冻精组15个周期、对照组100个周期分别注射MⅡ期成熟卵子163、1 157个,受精率冻精组显著高于对照组(84.05%vs73.29%,P<0.05),种植率、临床妊娠率则两组间差异无显著性(23.07%vs15.73%;53.33%vs37.00%,P>0.05),新生儿出生体重差异亦无显著性(P>0.05)。冻精组共妊娠8例,已分娩5例,继续妊娠3例。对照组妊娠37例,已分娩30例,1例死胎;继续妊娠3例;流产4例。两组均未出现重大的妊娠并发症及新生儿畸形。结论:采用PESA冷冻精子ICSI是治疗男性无精子症的一种经济、有效、安全的方法;但PESA冻精复苏率有待于进一步提高。