精子DNA甲基化的研究进展

DNA甲基化是表观遗传学研究内容之一,DNA甲基化在精子发生过程对基因的表达发挥着重要的调控作用,因此了解精子发生过程中DNA甲基化的变化对精子发生障碍研究有着至关重要的价值。本文对近年来精子的DNA甲基化信息进行综述,并对辅助生殖技术所带来的遗传风险作简要探讨。     

精子形态对体外受精胚胎移植妊娠结局的影响

目的：分析男性精液参数与卵裂率、优质胚胎率及妊娠率的相关性,探讨体外受精-胚胎移植（IVF-ET）周期中精子形态对精子功能的影响。方法：分析82个受精率100%的IVF-ET周期。以Diff-Quickstain方法染色,密度梯度离心法处理精液,WHO精子形态学标准评估精子形态,精子正常形态率≥14%为正常组,〈14%者为异常组。观察精液处理前后精子参数变化,比较妊娠组与未妊娠组的年龄、移植日子宫内膜厚度和精液各参数的情况。比较精液处理前后精子正常形态率与两组的正常受精率、卵裂率、优质胚胎率和妊娠率。结果：①密度梯度离心法处理后82例精液密度明显降低,活动力明显提高,差别有统计学意义。②精液处理前精子正常形态率：正常组11例（13.41%）,异常组71例（86.59%）;处理后正常组34例（41.46%）,异常组48例（58.53%）。异常组精液处理后正常精子形态的比例提高,中位数（P25,P75）分别为8.15%（6.38%,10.50%）vs.12.41%（9.98%,18.58%）,差别有统计学意义（P〈0.01）。③精液处理前精子正常形态率正常组和异常组的密度和活动力差别无统计学意义（P〉0.05）。④精液处理前后精子正常形态率正常组和异常组的妊娠率和优质胚胎率差异均无统计学意义（P〉0.05）;精子正常形态率正常组的卵裂率高于异常组（P〈0.05）。⑤妊娠组和未妊娠组除男女双方年龄差异有统计学意义外,移植日子宫内膜厚度和各精液参数差异均无统计学意义。结论：精子形态对妊娠结局无预测,密度梯度离心法能提高正常精子形态百分数,需根据精液处理后的结果选择受精方式。     

精子形态对体外受精-胚胎移植妊娠结局的影响

目的:分析男性精液参数与卵裂率、优质胚胎率及妊娠率的相关性,探讨休外受精一胚胎移植(IVF-ET)周期中精子形态对精子功能的影响.方法:分析82个受精率100%的IVF-ET周期.以Diff-Quickstain方法染色,密度梯度离心法处理精液.WHO精子形态学标准评估精子形态,精子正常形态率≥14%为正常组,<14%...     

精子与卵细胞膜上糖复合物在生殖过程中的作用

<正> 生殖过程的每一个环节,几乎都涉及到生殖细胞表面糖组分的变化,这些变化已用免疫分析、放射标记、糖苷酶降解及糖抑制实验等方法测定出来,特别是用具有糖专一性的凝集素研究精、卵细胞表面的糖复合物,以了解生殖过程中糖组分的变化,进而探索精、卵相互     

兔卵母细胞胞质内单精子注射后二次辅助激活对兔卵母细胞发育能力的影响

目的 观察兔卵母细胞胞质内单精子注射(ICSI)后2次辅助激活对兔卵母细胞发育能力的影响.方法 采集兔卵母细胞500枚,玻璃化冷冻后解冻,行ICSI后培养1 h,将存活的156枚卵母细胞分为10634激活1次组(30枚,加入浓度为5 μmoL/L的钙离子载体10634激活5 min)、氯化锶激活1次组(26枚,加入浓度为10 mmol/L的氯化锶激活20 min)、10634激活2次组(33枚,加入浓度为5μmoL/L的钙离子载体10634激活5 min后培养30 min,采用同样方法再次激活)、氯化锶激活2次组(28枚,加入浓度为10 mmol/L的氯化锶激活20 min后培养30 min,采用同样方法再次激活)和对照组(39枚,未加入任何激活剂)共5组,观察各组卵母细胞的受精率、卵裂率和囊胚形成率.结果 各组兔卵母细胞的受精率、卵裂率和囊胚形成率,氯化锶激活1次组分别为54%、27%和8%,均高于10634激活1次组(分别为33%、17%和3%)和对照组(分别为33%、18%和3%),但差异均无统计学意义(P>0.05);10634激活2次组分别为82%、55%和15%,均高于10634激活1次组,差异均有统计学意义(P<0.05);氯化锶激活2次组的受精率(61%)高于氯化锶激活1次组,卵裂率和囊胚形成率(分别为21%和7%)均低于氯化锶激活1次组,但差异均无统计学意义(P>0.05);10634激活2次组的受精率、卵裂率和囊胚形成率均高于氯化锶激活2次组,差异均有统计学意义(P<0.05).结论 冻融的兔卵母细胞ICSI后行2次辅助激活,可能会提高卵母细胞的受精率和胚胎早期的发育能力,但是激活后的卵母细胞分裂速度较快.     

精子DNA完整性的检测及其在辅助生殖技术中的作用

辅助生殖技术(ART)中,卵胞浆内单精子注射(ICSI)的出现开创了治疗男性不育的新局面,同时使得精子DNA完整性倍受人们关注。精子DNA完整性对自然受孕、人工授精、胚胎、胎儿、婴儿乃至成人的发育至关重要。研究表明精子DNA完整性是优于常规精液分析的独立参数,与ART结局有很高的相关性。因此精子DNA完整性检测对于ART有很重要的意义。目前有多种方法检测精子DNA完整性。     

微流控芯片优选人类精子研究

目的:探讨微流控芯片技术对精子的优选能力。方法:自行设计和制造微流控芯片,利用芯片对40例人精液标本进行精子分选实验,优化其分选条件,观察芯片处理前后精液各参数变化。同时对其中30例精液标本(A组:a+b级精子<20%组,n=15;B组:a+b级精子≥20%组,n=15)同时用芯片法和密度梯度离心法分选,比较2种方法分离前后精子活力、形态等参数的变化。结果:①优选后精子活力和精子正常形态率都可见显著提高(P<0.001;P<0.01)。②在精子活动力优选上A、B组芯片法均明显优于密度梯度离心法(P<0.01),尤其在A组这种优势更为明显(P<0.001)。而在精子形态优选上,2种方法无显著差异(P>0.05)。结论:微流控芯片技术在优选精子中具有较高的分选效率,且具有操作简单、分选时间短,对精子损伤小的特点,在辅助生殖技术中特别是体外受精中将有良好的应用前景。     

212个周期卵胞质内单精子显微注入术的临床效果

目的 :分析本中心 2 1 2周期卵胞质内精子显微注入 (intracytoplasmic sperm injection,ICSI)的结果。方法 :采用短方案超排卵后 ,分析其得卵率 ,ICSI后的卵裂率、妊娠率及妊娠率与女方年龄的关系。结果 :超排卵后每个周期的得卵率 1 0 .6个 ,ICSI后的卵裂率为 87.0 4% ,妊娠率为 32 .55% ,妊娠结果为 6例流产 ,6 3例分娩 ,获新生儿 74个。此外 ,妇女年龄与ICSI成功率有关。结论 :男方因素不孕夫妇用 ICSI及胚胎移植后 ,能获得较好的妊娠效果 ,女方年龄与 ICSI妊娠成功率有关     

辅助生殖技术中预防活性氧对精子的损伤

不育患者精子质量与生殖结局密切相关。活性氧(ROS)在精子获能、精卵融合等众多生理过程中扮演重要角色,但若产生过量或抗氧化能力不足,可在男性生殖系统中引发病理损伤,这些已在男性不育症中得到证实。本文就辅助生殖技术中活性氧的产生、预防问题作一综述。     

与精子功能相关抗原的分析

与精子功能相关抗原的分析贲昆龙（中国科学院昆明动物研究所，昆明，６５０２２３）精子的发生和形成始于睾丸的精小管，在精小管内，用原细胞逐渐分化成精母细胞、精细胞和精子。这个过程是在哺乳动物和人的免疫系统成熟之后发生的，是在由支持细胞形成的血睾屏障的严密...     

卵母细胞胞质内单精子注射的形态学观察

目的 了解卵母细胞胞质内单精子注射（ｉｎｔｒａｃｙｔｏｐｌａｓｍｉｃ ｓｐｅｍｒｍ ｉｎｊｅｃｔｉｏｎ,ＩＣＳＩ）过程中配子及卵裂球期胚胎各种异常形态与ＩＣＳＩ结果的相关性。方法 对１９９８年１１月至１９９９年８月在我中心接受ＩＣＳＩ治疗的４３个周期进行观察、分析。促性腺激素释放激素激动剂降调节长方案进行超促排卵,ＩＣＳＩ常规操作,比较配子及卵裂球期胚胎正常与各种异常形态同ＩＣＳＩ结果的相关性。结     

卵母细胞单精子显微注射治疗男性因素及不明原因不育

目的：研究应用卵母细胞单精子显微注射（ｉｎｔｒａｃｙｔｏｐｌａｓｍｉｃｓｐｅｒｍｉｎｊｅｃｔｉｏｎ，ＩＣＳＩ）技术，治疗因严重男性因素及不明原因引起不育患者的价值。方法：于１９９４年１２月，应用ＩＣＳＩ技术治疗３１例严重少、弱、畸精症及不明原因不育夫妇。按常规超排卵治疗，经阴道Ｂ超介导取卵，行成熟卵母细胞单精子显微注射受精。结果：２３７个成熟卵母细胞中，２１３个卵子存活，其中正常受精卵子１３２个，正常受精率为６２．０％。至１９９６年４月３０日，已有８例临床妊娠，临床妊娠率为２５．８％，其中１例在１９９６年１０月３日，于妊娠３９周＋３顺利分娩一婴儿。结论：ＩＣＳＩ技术用于治疗严重少弱畸精症及不明原因不育夫妇是适宜的。     

Sperm chromatin stability and its relationship with fertilization rate after Intracytoplasmic Sperm Injection (ICSI) in an assisted reproduction program

Objective Evaluate sperm chromatin stability and its relationship with the rate of fertilization after procedures of intracytoplasmic sperm injection (ICSI) in a program of assisted reproduction. Design Prospective study. Setting Institute of Gynecology and Reproduction. Patients Thirty-three women with their respective partners (12 couples in the study group and 21 couples in the control group) participating in a program of assisted reproduction. The study group was defined as men with >30% of non-decondensed spermatozoa (high sperm chromatin stability). Interventions A part of each seminal sample was used to evaluate sperm chromatin stability under SDS and EDTA treatment and the second aliquot was used for the ICSI procedure. Fertilization was evaluated 16–18 h post sperm injection at a pronuclear stage. The fertilized oocytes were further cultured for 24–48 h before transfer to the patient. Main outcome measures Fertilization rate. Results Thirty-five oocytes (35.7%) in the group of study and 109 oocytes (78.9%) in the control group showed two pronuclei (P < 0.001). The coefficient of determination between the SDS + EDTA (Grade 2) and rate of fertilization was r ² = 0.85 (P < 0.001) and the coefficient of regression was 1.72 ± 0.19 (β ± ES) (P < 0.001). Conclusions High sperm chromatin stability is a factor which reduces the rate of fertilization after ICSI procedure.     

Relationship Between the Sperm Motility Index Assessed by the Sperm Quality Analyzer and the Outcome of Intracytoplasmic Sperm Injection

Purpose: Intracytoplasmic sperm injection (ICSI) has been validated as a useful treatment in severe male-factor patients who could not achieve fertilization and live births by conventional in vitro fertilization treatment. To examine the impact of male factors on ICSI outcome, clinical laboratory data were retrospectively analyzed. Methods: One hundred two cycles of ICSI treatment indicated by severe male-factor infertility were entered into this study. Sperm parameters including sperm motility, sperm concentration, and sperm motility index assessed by the Sperm Quality Analyzer were evaluated. Results: Five hundred seventy-six metaphase II oocytes retrieved were manipulated. The normal fertilization (2 PN) rate per oocyte was 64.9 ± 26.0% (mean ± SD). Of the 99 transfers, 31 clinical pregnancies were obtained, yielding an average pregnancy rate of 31.3% per transfer. The mean sperm motility, sperm concentration, and sperm motility index were 20.3 ± 16.1% (range, 0 to 50%), 18.2 ± 25.1 × 10 ⁶ /ml (range, <1 to 150 × 10 ⁶ /ml), and 31.2 ± 45.0 (range, 0 to 220), respectively. Sperm concentration did not have a significant impact on fertilization rate by ICSI. In four cases, ICSI was performed using totally immotile sperm and the fertilization rate was 43.5%, which was significantly lower than that of some of the other sperm motility groups, and no pregnancy could be achieved. In 14 cases in which the sperm motility index assessed by the Sperm Quality Analyzer was 0, the fertilization rate (50.0%) was significantly lower than in most of the other sperm motility index groups. Conclusions: These findings suggest that in severe malefactor cases with totally immotile sperm or a sperm motility index of 0, the selection of good-quality sperm should be verified before injection.     

卵母细胞单精子显微注射治疗男性因素及不明原因不育156个周期临床分析

目的阶段性回顾应用卵母细胞单精子显微注射（ＩＣＳＩ）技术治疗严重男性因素及不明原因不育的临床效果。方法收集１９９５年１２月２０日至１９９８年１月１６日,在我院生殖医学研究中心接受ＩＣＳＩ技术治疗的严重男性因素及不明原因不育患者１４０对（１５６个周期）为研究对象,以１９９７年５月２８日为分界点,将其前的７４个周期划为第一阶段,其后的８２个周期为第二阶段,分析比较两阶段的临床结果。采用常规超排卵方案治疗,成熟卵母细胞行单精子显微注射。没有无核碎片及无核碎片＜２０％为Ａ类胚胎。结果两个阶段的成熟卵母细胞数和卵裂率差异无显著性,但注射后存活卵子数、正常受精数、卵裂球的质量和临床妊娠率,第二阶段分别为７０３个（８９５％）、４８７个（６９３％）、Ａ类胚胎占７４２％和３６６％,较第一阶段的５９２个（８５１％）、３８８个（６５５％）、Ａ类胚胎占６６０％和２７０％,有显著性提高。结论ＩＣＳＩ的操作直接影响胚胎质量,随着ＩＣＳＩ技术的不断成熟和完善,其在治疗男性因素及不明原因不育中的作用是肯定的     

Diamour®, a newly vitrification device for human blastocysts,provides the same efficient perinatal outcomes as the commonly used Cryotop®

ObjectiveThe purpose of this study was to investigate the effectiveness of Diamour®, a newly developed vitrification device, for the ultra-rapid frozen storage of human blastocysts.Materials and methodsSurplus blastocysts obtained using assisted reproductive technology at our clinic during a 2-year period (2018–2019) were assigned for vitrification with the Diamour® device in 2019 or a comparator device in 2018. We retrospectively compared outcomes (clinical pregnancy rate, abortion rate, live birth rate, and perinatal outcomes) between the two vitrification devices.ResultsWe obtained 228 blastocysts from 178 patients, and 118 blastocysts were cryopreserved using the Diamour® vitrification device and the other 110 were cryopreserved using a Cryotop®, the comparator device. We found no significant difference between the Diamour®-vitrified and comparator-vitrified blastocysts in clinical pregnancy rate (32.2% vs. 30.9%; p = 0.83), abortion rate (17.0% vs. 9.1%; p = 0.08), and live birth rate (25.4% vs. 26.4%; p = 0.87). The Diamour®-vitrified blastocysts yielded 30 live births, and the comparator-vitrified blastocysts 29. The two devices yielded no significant difference in birth weight (Diamour®: 3049 ± 337 g, comparator: 3008 ± 340 g; p = 0.65) and congenital abnormalities, with just 1 case (cleft lip) noted with Diamour® vitrification (p = 0.33). The devices also showed no significant differences in the incidence of gestational diabetes mellitus, hypertensive disorders of pregnancy, placenta previa, preterm birth, macrosomia, low birth weight, and delivery method.ConclusionThe Diamour® device achieves outcomes similar to those of the currently widely used Cryotop® device for clinical pregnancy with frozen-thawed human blastocysts.     

Preimplantation sex selection demand and preferences among infertility patients in Midwestern United States

Purpose To determine the demand and preferences of infertility patients for sex selection for nonmedical reasons, and to investigate the relation between these choices and their demographic and socioeconomic characteristics. Methods A cross-sectional, self-administered survey by mail was conducted at a University hospital-based fertility center of 1,350 consecutive women who presented for infertility care, to assess patient demand and preferences for sex selection. Results Of respondents, 49% wanted to select the sex of their next child for no added cost. Of these patients, 56% had no living children and 37% had children all of one sex. After adjustment for observed predictors of gender preference, we found a significant preference for a female child among women who had only sons, had more living children, or were single. Nulliparous women did not significantly prefer one sex over the other. Among parous women, those with only daughters significantly desired to select a male child, whereas those with sons significantly desired to select a female child. Conclusion There is significant demand among infertility patients for preimplantation sex selection, with a significant portion of this demand coming from patients who do not have any children or have children all of one sex. Presented in part at the American Society for Reproductive Medicine (ASRM) Annual Meeting, New Orleans, Louisiana, October 2006. Financial support: none Conflict of interest: none     

Very Low Sperm Count Affects the Result of Intracytoplasmic Sperm Injection

Purpose: The aim was to examine the influence of extremelylow sperm count on intracytoplasmic sperm injection(ICSI) outcome. Methods: Over 1000 consecutive unselected ICSI cycleswere divided into four groups according to spermconcentration of their patients: A, cryptozoospermia, 107 patients; B,sperm concentration of 1×10⁴, 146 patients; C, spermcount of 1×10⁴–1×10⁵, 135 patients; and concentration of 1×10⁵ and < 10×10⁶/ml (control group), 688 patients. Results: A significant decrease in pregnancy rate wasnoticed in the cryptozoospermic group in comparison to thecontrol group (20% vs. 31%). Fertilization rate in group Awas significantly lower in comparison to all other groups,respectively (46% vs. 52%, 54%, 61%). Embryo quality wasinferior in group A in comparison to the control group. Ahigher yet not statistically significant abortion rate wasobserved in the cryptozoospermic group (as well as in groupC) (30%, 27%) compared to the control group (15%). Conclusions: It seems that an extremely low sperm counthas a negative effect on the outcome of ICSI. Neverthelesspatients with cryptozoospermia should not be offered ICSItreatment with the ejaculated sperm before karyotype isestablished.     

Overview Analysis of Outcomes with Intracytoplasmic Sperm Injection

The study of male factor infertility only recently has begun to receive increasing attention because there are now more therapeutic options. This situation has become possible because of the availability of assisted fertilization techniques, particularly intracytoplasmic sperm injection (ICSI) which has resulted in high fertilization and pregnancy rates. Although the sperm for ICSI have been used mainly from ejaculated specimens, it is now possible to use sperm from epididymal and testicular sources. Factors that affect the pregnancy rate with ICSI include female age, the number of embryos transferred, and oocyte morphology. The small follow up studies of children born after ICSI provide reassurance that the technique appears to be safe.     

轻度子宫内膜异位症患者的腹腔液对精子运动及超微结构的影响

目的:观察轻度子宫内膜异位症(EM)不孕患者的腹腔液对精子运动及超微结构的影响。方法:经过处理的精子在体外分别和轻度EM不孕患者、正常妇女的腹腔液及精子培养液孵育5小时,观察其运动参数及超微结构的变化;酶联免疫法测量EM腹腔液肿瘤坏死因子α(TNF-α)的含量。结果:EM组腹腔液TNF-α含量高于正常妇女(P<0.05);EM组精子平均直线速度(VSL)、平均曲线速度(VCL)、平均路径速度(VAP)、快速直线运动精子活率(MOT)4项运动指标均低于正常组和培养液组(P<0.05),与TNF-α含量呈负相关(P<0.05),EM组精子头部、顶体及尾部主段胞质肿胀、胞膜破损,而孵育前及孵育后另外两组精子结构无改变。结论:轻度EM不孕患者腹腔液TNF-α值升高,经体外观察腹腔液对精子运动及精子形态学均有损害作用,推测这些可能是EM患者不孕的原因之一。