精液优化处理后DNA碎片指数与IVF-ET胚胎质量及妊娠结局的关系

目的:评估经密度梯度离心联合上游法优化处理后的精子DNA碎片指数(DFI)与辅助生殖体外受精-胚胎移植(IVF-ET)胚胎质量及妊娠结局的关系。方法:回顾性分析2022年4月至12月在郑州大学第一附属医院生殖医学中心因单纯输卵管因素行IVF-ET的257个周期,比较男方精液优化前后精液参数和精子DFI;按优化后精子DFI将其分为高DFI组(DFI>5)与低DFI组(DFI≤5),比较两组的胚胎发育及妊娠结局;根据临床妊娠情况分为妊娠组与非妊娠组、持续妊娠组及早期流产组,比较精子DFI情况。结果:与处理前相比,精液优化处理后前向运动精子及正常形态精子百分比提高,精子DFI下降(P<0.001)。精子优化处理后高DFI组的早期流产率高于低DFI组(P<0.05)。105例临床妊娠周期中早期流产组精液优化处理前、后DFI均高于持续妊娠组(P<0.05)。结论:密度梯度离心联合上游优化处理是一种有效的精液制备方法,可提高精子前向运动能力和正常形态精子比例,降低精子DFI;优化后精子高DFI可能增加临床妊娠后早期流产的风险。

Where is the rudder of a fish?:the mechanism of swimming and control of self-propelled fish school

Numerical simulation and control of self- propelled swimming of two- and three-dimensional biomimetic fish school in a viscous flow are investigated. With a parallel computational fluid dynamics package for the two- and three-dimensional moving boundary problem, which combines the adaptive multi-grid finite volume method and the methods of immersed boundary and volume of fluid, it is found that due to the interactions of vortices in the wakes, without proper control, a fish school swim with a given flap- ping rule can not keep the fixed shape of a queue. In order to understand the secret of fish swimming, a new feedback con- trol strategy of fish motion is proposed for the first time, i,e., the locomotion speed is adjusted by the flapping frequency of the caudal, and the direction of swimming is controlled by the swinging of the head of a fish. Results show that with this feedback control strategy, a fish school can keep the good order of a queue in cruising, turning or swimming around circles. This new control strategy, which separates the speed control and direction control, is important in the construction of biomimetic robot fish, with which it greatly simplifies the control devices of a biomimetic robot fish.

Effect of Swim-Up and Percoll Treatment on Sperm Quality and In vitro Embryo Development in Yak

This study was designed to determine the effect of different sperm preparation treatments on yak sperm quality and in vitro embryo development.Frozen-thawed semen samples were treated using swim-up or percoll gradient centrifugation methods.Sperm concentration,progressive motility,recovery of motile sperm,membrane integrity,acrosome and chromatin integrity were scored and compared in recovered samples and controls.In addition,the effects of two sperm separation treatments on embryos capable of cleavage and in vitro development to the blastocyst stage were evaluated.Swim-up separated sperm showed a higher motility,while the concentration of spermatozoa recovered and percent recovery of motile sperm were higher with percoll gradient centrifugation separation.According to the optical and electron microscopies,swim-up produced higher percentage of sperm with intact plasma membrane and acrosome than percoll gradient centrifugation separation.However,there was no difference in the percentage of sperm with intact chromatin between two treatment groups.Cell numbers in the blastocysts of two groups were not different.The blastocyst rate was similar in both groups,whereas cleavage rate was higher when swim-up was used.

梯度密度离心法与上游法在冷冻精液供精体外受精-胚胎移植术中应用比较

目的:观察在使用冷冻精液的供精体外受精-胚胎移植术中采用梯度密度离心法与上游法处理精液后的妊娠率,比较两种精液处理方法对体外受精-胚胎移植术成功率的影响。方法:回顾本院常规体外受精-胚胎移植术中使用冷冻供精71个周期,其中上游法35周期,梯度密度离心法36周期的资料,观察经上游法和梯度密度离心法处理后精子密度、精子总活动率、前向运动精子活动率等精液情况和受精率、卵裂率、临床妊娠率。结果:梯度密度离心组在精子密度、精子总活动率、前向运动精子活动率3项指标均高于上游组,而受精率、卵裂率、临床妊娠率也是梯度密度离心组较上游组高(P<0.01)。结论:在使用冷冻精液的供精体外受精-胚胎移植术中,采用梯度密度离心法处理精液优越于上游法,能获得更高的受精率和妊娠率。

精液优选对精子DNA碎片的影响及其对体外受精-胚胎移植妊娠结局的影响

目的评估密度梯度离心联合上游法改善精子DNA碎片(sperm DNA fragmentation)的效率及其对体外受精-胚胎移植(IVF-ET)妊娠结局的影响。方法收集120份处理前后的精液,用精子染色质扩散法检测精子DNA碎片,评估其对100个新鲜移植患者妊娠结局的影响。结果优选后,男性不育组精子DNA碎片率(SDFI)由22.75(14.44,30.25)降至11.5(5.60,22.79),对照组则由20.86(15.00,26.81)降至7.5(3.63,15.44);妊娠组精子DNA碎片明显低于非妊娠组,ROC曲线下面积是0.667;高水平组(DFI≥20%)的妊娠率和出生率低于低水平组(DFI<20%),差异有统计学意义。结论密度梯度离心联合上游法可减少精子DNA碎片;优选后DNA碎片对IVF-ET妊娠结局具有预测价值,一旦超过阈值,虽可正常受精,但妊娠率和出生率下降。

Comparative study on efficacy of three sperm-separation techniques

Aim: To evaluate the comparative effectiveness of real-time sperm separation technique (Wang's tube method) andother two conventional methods in isolating high-quality sperm preparation, and to compare the spouse pregnancy ratein intrauterine insemination (IUI) with sperm preparations isolated by these methods. Methods: The effectivenessof the real-time sperm separation technique, the conventional swim-up and the Percoll discontinuous density gradientmethods in isolating sperm preparations from 60 infertile patients (20 with apparently normal semen and 40, abnormalsemen contaminated with microorganisms and other impurities) was evaluated and compared. The microorganisms to beremoved included bacteria, vires, Chlamydia trachomaticum, Ureaplsama urealyticum, etc. The spouse pregnancyrates in IUI with sperm preparations isolated by these three techniques from 80 oligoasthenoteratospermic patients werealso compared. Results: The quality (including the percentages of normal form, normal-chromatin and motilesperm, and the grade of motility) of sperm obtained by the real-time sperm separation technique was much higher ( P< 0.01) as compared with those by the other two methods. The Wang's tube method was also more effective in remov-ing microorganisms and other impurities. The method provided a higher IUI pregnancy rate than the other two spermseparation techniques ( P < 0.05). Conclusion: The real-time sperm separation technique is the most effectivemethod so far available in isolating high-quality sperm samples to be used in assisted reproduction.

The ability of sperm selection techniques to remove single- or double-strand DNA damage

A wide variety of techniques for the preparation of sperm are currently available, of which the most commonly employed are densitygradient centrifugation （DGC） and swim-up （SUP）. To date, these methods appear to be effective in selecting functional sperm for assisted reproduction techniques （ART）, but they may have negative effects on sperm DNA. In this study, the ability of these semen processing techniques to eliminate spermatozoa containing single- and double-strand DNA damage was assessed by the two-tailed comet assay and the sperm chromatin dispersion test in 157 semen samples from patients seeking assisted reproduction treatment. Our results indicated that SUP and DGC are equally efficient in eliminating spermatozoa containing double-strand DNA damage and sperm with highly damaged （degraded） DNA, as characterized by the presence of both single- and double-strand DNA breaks. However, DGC is more efficient than SUP in selecting spermatozoa that are free from single-strand DNA damage. Future studies should characterise the importance of the various types of DNA damage and examine the sperm processing protocols used in each laboratory to determine their ability to eliminate DNA damage and hence, prevent the potential transmission of genetic mutations via ART.

改良精子上游法用于体外受精-胚胎移植

本文报告一种用以收集高活动力精子的改良上游枝术,并在体外受精-胚胎移植中与常规洗涤法相比较。用上游法获得的精子悬液平均精子活动力为93.36±8.44%(X±SD),Ⅰ、Ⅱ级前向运动精子比例占75%以上。比较两种精子分离方法的体外受精率:上游法为45.65%,洗涤法为30.30%,p<0.05。在8例以上游精子授精所得胚胎行胚胎移植中,2例获得妊娠并已足月顺产出“试管婴儿”。

微流控芯片精子分选法对精子常规参数及DNA完整性的影响

目的:研究微流控芯片精子优选技术对精子常规参数及DNA完整性的影响。方法:自行设计制作微流控芯片,利用芯片处理技术和上游法对40例精液标本进行精子优选,通过计算机辅助精液分析系统及染色质扩散试验从精子常规参数及DNA完整性两个方面评价精液体外处理对精子的影响。结果:精液经微流控芯片法和上游法处理后,精子活力、精子正常形态率以及精子尾部肿胀率均有显著提高(P<0.01),精子的DNA损伤率明显降低(P<0.01)。微流控芯片法与上游法相比,优选后前者精子DNA损伤率明显低于后者[(8.4±5.8)%vs(16.4±9.2)%,P<0.01],而其他参数差异无显著性。结论:微流控芯片技术在精子优选中能获得精子DNA损伤程度小的高质量精子。

密度梯度离心法与直接上游法优化处理精液对宫腔内人工授精临床妊娠率的影响

目的:应用密度梯度离心法和直接上游法处理不明原因不孕和轻度精液异常患者的精液,比较两种方法对宫腔内人工授精(IUI)临床妊娠率的影响。方法:回顾性分析2013年1月至2014年12月期间,在我中心进行促排卵+IUI的571个周期患者,统计其精液优化处理方法和临床妊娠率。结果:总体临床妊娠率为密度梯度离心法(17.8%)显著高于直接上游法(11.4%)(P<0.05);不明原因不孕组的临床妊娠率为密度梯度离心法(33.7%)显著高于直接上游法(20.3%)(P<0.05);轻度精液异常组的临床妊娠率为密度梯度离心法(11.6%)显著高于直接上游法(7.6%)(P<0.05)。结论:密度梯度离心法相对直接上游法能有效提高因不明原因不孕和轻度精液异常患者IUI的临床妊娠率。

精子分离两种方法的比较和评价

目的 :探讨王氏管法和精子上游法在精子分离中的应用 ,比较两者的优缺点。 方法 :随机选择 12 7份精液 ,采用 2种方法进行配对处理 ,比较 2种方法分离前后精子活动率、形态、活力和精子顶体形态、精子染色质以及精子膜完整性等参数的变化。 结果 :2种方法处理后的精子活动率、精子活力以及精子形态有显著的差异 (P <0 0 1) ,王氏管法处理后精子顶体完整率、正常染色质率及精子膜完整率亦显示出显著的差异 (P <0 0 1)。 结论 :王氏管法和精子上游法在精子处理中均可获得较好的精子质量 ,但王氏管法则获得更优质的精子 ,且具有方便操作和实用的特点。

荧光原位杂交检测精子上游时间对XY精子比例的影响

目的：运用荧光原位杂交（fluorescent in situ hybridization，FISH）技术检测精子改良上游过程中，不同上游时间对含X／Y精子的分离作用，探讨较长上游时间跨度（0～150分钟）对上游后XY精子比例的影响。方法：分析从志愿者取得的正常精液标本10份，每份均进行精子不同时间分批改良上游、固定和荧光原位杂交分析，计数分析X精子占XY精子总数的比例。结果：杂交区域内精子总杂交率为98．33％。未上游时X精子比例为平均为50．03％。通过SPSS方差分析：分时间段检测的X精子占XY精子总数的比例与未上游时差异无显著性（P〉0．05），不同组间差异无显著性（P均〉0．05）。结论：改良上游法对XY精子的分离没有显著效果，没有直接证据表明人工授精和体外受精．胚胎移植过程中精子的上游处理会影响到出生婴儿的性别比例。

不孕症患者宫腔内人工授精精子优选方法分析

目的:比较分析精子不同优选方法与宫腔内人工授精(IUI)成功率的相关性,以提高IUI妊娠率。方法:回顾性分析本院生殖中心门诊不孕症患者452例671个IUI周期,按精子优选方法分为3组:含5%人血清白蛋白(HSA)的Earle's液上游法、SpermR inse上游法、SupraSperm密度梯度离心法。比较各组患者的处理前后精子密度与活动率及临床妊娠结局。结果:3种精子优选方法处理后精子活动率及a+b级精子百分率显著增加(P<0.01)。含5%HSA的Earle's液上游法221个IUI周期,妊娠21个周期,周期临床妊娠率9.5%;SpermR inse上游法215个IUI周期,妊娠34个周期,周期临床妊娠率15.8%,与含5%HSA的Earle's液上游法比较有显著性差异(P<0.05);SupraSperm密度梯度离心法235个IUI周期,妊娠34个周期,周期临床妊娠率14.5%,与含5%HSA的Earle's液上游法比较无统计学差异(P>0.05)。结论:SpermR inse上游法的IUI临床妊娠率明显高于含5%HSA的Earle's液上游法,适用于各类不孕症患者。SupraSperm密度梯度离心法也具有较高的周期临床妊娠率,洗涤效果好,操作便捷高效,尤其适用于含炎症细胞、死精子和畸形精子等不良成分较多的精液。

刺五加水提物对人精子体外运动活力的影响

目的研究刺五加水提物对正常男性精子体外运动活力的影响,探讨其可能的作用机制。方法将20例正常男性的精子通过上游优化法处理后,一组精子加入10 mg.mL-1的刺五加水提物,另一组加入等量的Ham’s F10培养液作对照。应用计算机辅助的精子分析系统,分别测定孵育0.5,1,2,3,4,5 h后人精子各运动参数。结果随着时间的延长,加入Ham’s F10培养液的精子活动能力有不同程度的下降;加入10 mg.mL-1的刺五加水提物后,不同时段精子的运动能力均较对照组明显提高,精子存活率、前向运动精子百分率、曲线运动速度、直线运动速度和平均路径速度与对照组比较,均明显改善(均P<0.05)。结论刺五加水提物能明显提高正常男性精子的体外存活率和运动能力。

日本竞技游泳崛起的原因分析

日本竞技游泳重新崛起的原因主要包括:社会和政府的重视,科研力量的增强,选拔制度的改革,教练员资格认定制度的完善,重视情报收集和分析,高学历的科技体育志愿者的支持,重视选手的精神锤炼和与教练员之间的交流。这些问题应引起我国游泳界备战北京奥运会的高度重视。

暑期游泳速成教学实践与研究

运用文献资料、问卷调查、实验比较和数理分析等方法对暑期游泳学习班速成教学法进行教学实践与研究,探索提高暑期游泳学习班的教学效果,以满足日益增长的游泳爱好者的需要,从而赢得市场,并促进游泳运动的普及化和大众化。

两种方法处理后精子形态的变化

目的研究上游法和密度梯度离心法(Percoll法)处理前后精子畸形率的改变。方法60例精液标本分别行上游法和密度梯度离心法优选,处理前后均行精子形态改良巴氏染色分析。结果上游法和密度梯度离心法都可以显著降低精子的畸形率,上游法以颈部和尾部畸形最为明显,其他畸形降低不显著;而密度梯度离心法处理后精子以大头和双头畸形降低较为显著,其他种类畸形精子率下降不显著。结论上游法和密度梯度离心法对改善精子畸形率的作用有限。

王氏管精子优选法的应用及效果

目的 探讨王氏管精子优选法在精子优选的应用 ,并与传统的精子上游法进行比较。方法 随机选择 5 7份精液 ,采用两种方法进行配对处理 ,比较两种方法分离前后、两者间精子活动率、形态以及精子顶体形态和精子染色质等参数的变化。结果 两种方法处理后的精子活动率、精子形态差异有显著性 (P <0 0 1) ,王氏管法处理后精子顶体完整率、正常染色质率亦显示出显著的差异 (P <0 0 1)。结论 王氏管精子优选法和精子上游法在精子处理中获得较好的精子质量 ,而王氏管精子优选法则获得更优质的精子 。

梯度离心在体外处理精子结合抗体中的应用

目的:比较单纯PureSperm方法和PureSperm联合上游法处理精子结合抗体阳性精子的效果。方法:采用单纯梯度PureSperm及PureSperm联合上游法处理精子结合抗体阳性的精子,比较2种处理方法处理后精子结合抗体消退情况。结果:2种方法处理后,精子结合抗体滴度较处理前降低(P<0.01);梯度PureSperm联合上游法处理后,精子结合抗体滴度的下降较单纯PureSperm方法更显著(P<0.05)。结论:采用梯度PureSperm离心或梯度PureSperm离心联合上游法处理精液,可以提高精子质量,再行宫腔内人工授精可能提高免疫性不育的治疗效果。

上游技术对精子运动参数的影响

目的了解精子各运动参数在上游法处理前后的变化。方法37例男性精液标本,经上游法处理后,应用计算机辅助精液分析仪分析精子各项运动参数。结果上游法处理后,以快速直线运动为主的前向运动精子百分率和曲线速度、直线速度等运动参数值均高于上游法处理前(P<0.001)。结论上游法处理精液后,快速运动精子和精子各项运动参数值均有改善。