乙肝病毒对人精液参数和精子DNA完整性的影响

目的:探讨精液中乙肝病毒存在对精液参数和精子DNA完整性的影响。方法:采用实时荧光定量PCR技术对153例血清乙肝表面抗原阳性患者精液进行乙肝DNA定量检测,采用精子染色质扩散法检测精子DNA损伤指数(DFI),对比分析精液HBV DNA阳性组(A组,n=43)与阴性组(B组,n=110)精液参数包括精液体积、精子浓度、存活率、前向运动精子百分率、平均直线速率(VSL)、平均路径速率(WAP)结果,以及HBV DNA拷贝数与精子DNA损伤指数的关系。结果:乙肝感染者精液HBV DNA检出率为28.1%(43/153);两组年龄、精液体积、精子浓度无统计学差异(P>0.05),A组精子存活率显著低于B组[(51.4±17.1)%vs(58.0±18.8)%,P>0.05],前向运动精子率显著低于B组[(24.5±10.1)%vs(29.6±13.3)%,P>0.05],VSL显著低于B组[(19.9±4.5)μm/s vs(23.7±4.0)μm/s,P<0.01],WAP显著低于B组[(23.4±5.3)μm/s vs(26.5±7.0)μm/s,P<0.01];A组精子DNA碎片指数显著高于B组[(24.2±9.4)%vs(19.3±8.0)%,P<0.01];精液中HBV DNA拷贝数与精子DNA碎片指数呈正相关(r=0.819,P<0.01)。结论:精液中HBV DNA存在对精子数量尚未构成显著影响,但可能改变一些精液参数,包括精子活力和运动速度以及精子DNA完整性,精液中乙肝病毒载量和精子DNA损伤之间呈现病毒载量-效应关系。     

非连续密度梯度法对弱精子症患者精子参数及DNA完整性的影响

目的通过非连续密度梯度法制备弱精子症患者精液,分析制备前后精液常规分析质量参数及精子DNA完整性,评估非连续密度梯度法在改善弱精子症患者精子质量方面的有效性。方法 26例弱精子症患者来源于本院门诊,禁欲2~7d后手淫取精,使用一次性精子计数玻片(goldcyto),采用SCA精子质量分析系统对精子制备前后精子浓度、精子总活力[前向运动精子(PR)+非前向运动精子(NP)]和PR百分率进行分析;采用改良巴氏染色法分析精子形态;同时运用精子吖啶橙染色方法对制备前后的精子DNA完整性进行检测。结果 26例精液标本制备前精液体积(4.22±1.54)mL,精子浓度(51.9±44.65)×10~6/mL,精子总活力(22.5±6.34)%,PR百分率为(17.3±5.5)%,正常形态精子百分率(6.58±2.50)%,精子DNA完整率(64.96±13.33)%;制备后精子悬液体积统一调至0.5m L,精子浓度(18.91±24.61)×10~6/mL,精子总活力(31.77±21.15)%,PR百分率(26.2±20.00)%,正常形态精子百分率(7.77±3.14)%,精子DNA完整率(72.23±12.43)%。统计结果提示精子制备前后精子浓度、精子总活力、PR百分率、正常形态精子百分率和精子DNA完整率均存在显著性差异(P0.05)。结论非连续密度梯度法制备精子能显著提高弱精子症患者精子总活力、PR百分率、正常形态精子百分率和精子DNA完整率,有效改善制备后的精子质量,为后续实施人类辅助生殖技术提供保障。     

精子CMA3阳性率与IVF受精率的相关性研究

目的探讨精子色素酶A3(CMA3)阳性率与IVF受精率之间的相关性。方法收集2015年4月至2016年7月因单纯输卵管性不孕在我院生殖医学科行常规IVF助孕的156个周期,以受精率<25%为低受精,根据受精率不同分为正常受精组(134个周期)和低受精组(22个周期)。取卵日,将授精后剩余的洗涤精液行精子浓度和活动率分析,并将精子进行CMA3染色。比较两组优选后的精子参数,分析IVF受精率与精子参数及CMA3阳性率的关系。结果与正常受精组比较,低受精组精子CMA3阳性率显著升高[(20.0±4.2)%vs.(30.7±2.3)%],优选后前向运动精子的百分率显著降低[(90.4±4.8)%vs.(74.3±3.4)%](P<0.05);两组精子浓度比较无显著性差异(P>0.05)。IVF受精率与优选后的前向运动精子百分率呈正相关(r=0.76,P<0.01)。精子CMA3阳性率与优选后前向运动精子百分率(r=-0.82,P<0.01)及IVF受精率(r=-0.83,P<0.01)呈显著负相关,与精子浓度则无显著相关性(P>0.05)。结论精子CMA3阳性率与IVF受精率负相关,但其具体机制尚需进一步研究探讨。     

3种精子计数池在计算机辅助精液分析系统中的质量评价

目的:应用计算机辅助精液分析(CASA)系统对3种常用精子计数池进行比较,评价其在精子密度及活力分析方面的差异。方法:用浓度为(20±5)×106/ml乳胶珠溶液模拟精液样本,以Makler计数池、Leja计数池、Microcell计数池3种精子计数池作为精液分析工具,每组测定30次,以CASA系统对其密度进行分析并统计均值(x±s)和变异系数(CV);同时收集54例门诊患者精液标本用3种精子计数池分别进行活力分析,计算各组计数池前向运动精子百分率及活动精子百分率。结果:Makler计数池、Leja计数池、Microcell计数池等3种精子计数池的密度均值和变异系数分别为(25.90±3.97)、(18.74±1.62)、(20.35±2.55)×106/ml和15.31%、8.64%、12.54%。3种精子计数池密度均值间均存在明显的差异(P<0.05);Makler计数池、Leja计数池、Microcell计数池3种精子计数池的前向运动精子百分率和活动精子百分率分别为(46.54±17.09)%、(30.65±14.88)%、(30.49±13.21)%和(59.75±16.12)%、(46.76±14.11)%、(43.11±14.02)%。Makler计数池的分析结果均明显高于Leja计数池、Microcell计数池(P<0.05),而Leja计数池与Microcell计数池比较则无明显差异(P>0.05)。结论:不同类型的精子计数池得到的密度结果会存在明显的差异,在进行精子活力分析时,Makler计数池获得的结果会较Leja计数池与Microcell计数池的结果偏高。     

复方氨基酸胶囊联合维生素E治疗特发性弱精子症的临床研究

目的:研究复方氨基酸胶囊联合维生素E治疗特发性弱精子症的疗效。方法:纳入2014年2月至2015年1月门诊120例弱精子症患者。患者被随机分为两组:治疗组70例,年龄23~43岁,平均32.5岁;对照组50例,年龄23~44岁,平均31.7岁。治疗组口服复方氨基酸胶囊+维生素E,对照组仅服维生素E,均连续服用90 d。比较治疗前、后精子活动力(PR+NP)、前向运动精子百分率(PR)和配偶妊娠率,评估其临床效果。结果:治疗前,治疗组和对照组PR+NP、PR分别为(26.24±6.56)%、(24.65±6.43)%和(15.13±5.68)%、(14.73±6.16),差异无统计学意义(P均0.05)。治疗90 d后,治疗组PR+NP、PR分别为(49.63±9.78)%、(33.33±5.64)%,与治疗前比较差异具有统计学意义(P0.05);对照组PR+NP、PR分别为(37.67±7.98)%、(27.23±6.46)%,与治疗前比较差异具有统计学意义(P0.05),但治疗组PR+NP、PR改善明显优于对照组,差异具有统计学意义(P0.05)。治疗组有4例(5.71%)患者配偶成功受孕,对照组有1例(2.00%)患者配偶成功受孕,总有效率治疗组(74.28%)明显高于对照组(44.0%)(P0.05)。两组均未出现不良反应。结论:复方氨基酸胶囊联合维生素E可改善特发性弱精子症患者精子活力。     

益精方治疗弱精子症的临床随机对照研究

目的:观察益精方治疗弱精子症的临床疗效。方法:450例患者以信封法随机分为治疗组(益精方组)300例和对照组(五子衍宗丸+左卡尼汀口服液组)150例,经精液常规检查诊为弱精子症,治疗组服用益精方,每天1剂,温水冲服,分2次服用,对照组服用五子衍宗丸+左卡尼汀口服液,五子衍宗丸9 g,口服,每日2次,左卡尼汀口服液,10 ml,口服,每日2次,均治疗3个月,于治疗前及治疗后的第1个月(第27~33天)、2个月(第57~63天)和3个月(第87~93天)分别检测患者的精液常规,比较治疗前后精子浓度、前向运动精子百分率(PR)、精子总活力(PR+NP)等精液参数的变化。结果:两组患者治疗前精液常规检查结果无显著差异(P0.05)。治疗组治疗1、2、3个月时,精子浓度分别为49.66±10.91、55.21±11.46、74.90±13.07(×10~6/ml),PR分别为23.81±2.56、26.12±2.34、32.17±1.62(%),PR+NP分别为34.17±3.43、36.59±3.36、47.08±2.97(%),精液体积分别为3.15±1.06、3.12±0.90、3.27±0.78(ml),液化时间分别为31.68±3.14、30.38±3.44、30.86±2.42(min),组内比较显示,治疗后精子浓度、PR、PR+NP较治疗前差异显著(P0.001),精液体积、液化时间差异不明显(P0.05)。对照组治疗1、2、3个月时,精子浓度分别为40.53±8.32、47.51±12.73、56.14±11.98(×10~6/ml),PR分别为25.17±2.64、27.23±2.25、31.89±2.27(%),PR+NP分别为33.89±2.26、37.38±4.79、40.35±3.06(%),精液体积分别为3.16±1.78、3.15±0.96、3.12±0.65(ml),液化时间分别为29.36±4.25、28.21±3.26、28.33±3.59(min),组内比较显示,治疗后精子浓度、PR、PR+NP较治疗前差异显著(P0.001),精液体积、液化时间差异不明显(P0.05)。组间比较显示,治疗组精子浓度及PR+NP增长率较对照组有显著差异(P0.001),PR增长率两组间差异不明显(P0.05)。结论:益精方可在生精周期多个阶段调控精子的发生,提高弱精子症患者的PR及PR+NP比例,是一种有效的治疗弱精子症的方剂。     

冷冻保护剂与精液比例对冷冻复苏后精子活动力的影响

目的:比较冷冻保护剂与精浆的不同比例对人类精子冷冻复苏后活动力的影响。方法:将57例志愿者的精液标本分别采用冷冻保护剂∶精液=1∶1和冷冻保护剂∶精液=1∶3的体积比进行冷冻,比较精子复苏后的前向活动率和总活动率。结果:冷冻前的前向运动精子百分率和总活动率分别为(58.60±5.57)%和(66.17±5.24)%。采用冷冻保护剂∶精液=1∶1比例进行冷冻复苏后的前向运动精子百分率和总活动率分别为(40.53±8.97)%和(51.23±9.30)%;采用冷冻保护剂∶精液=1∶3比例进行冷冻复苏后的前向运动精子百分率和总活动率分别为(44.70±8.67)%和(51.50±7.40)%。冷冻前后精子的前向运动百分率和总活动率差异有显著性(P<0.05)。两种不同比例的保护剂相比冷冻后前向运动精子百分率差异有显著性(P<0.05),而总活动率差异没有统计学意义。结论:冷冻对精子活动力损伤明显,冷冻保护剂∶精液=1∶3比例较冷冻保护剂∶精液=1∶1比例可提高冷冻后前向运动精子百分率。     

男性年龄与精子顶体酶活性及精子DNA碎片指数的相关性分析

目的探讨男性年龄与精子顶体酶活性、精子DNA碎片指数(DFI)的相关性。方法选取2016年1~8月在我院生殖医学中心就诊的436例不育症男性患者为研究对象,所有患者均行精液常规检查、精子顶体酶活性检查和(或)精子DFI分析。将患者按年龄分为<30岁、30~39岁、≥40岁3组,分析各组的精液常规、顶体酶活性及精子核DFI的差异及其相关性。结果不同年龄段患者的体重指数(BMI)、禁欲天数、精液量无显著性差异(P>0.05);年龄≥40岁组患者的前向运动精子百分率、活动精子百分率及精子顶体酶活性显著低于<30岁和30~39岁组(P<0.05);≥40岁组患者的精子DFI显著高于<30岁和30~39岁组(P<0.05)。年龄与前向运动精子百分率及活动精子百分率之间呈负相关(P<0.05),但是相关性较弱。精子顶体酶活性与精子正常形态率、前向运动精子百分率、非前向运动精子百分率、活动精子百分率呈正相关(P<0.05);精子DFI与年龄、禁欲天数、前向运动精子百分率呈正相关(P<0.01),与精液量、精子浓度、活动精子百分率呈负相关(P<0.05);精子顶体酶活性和DFI之间无相关性(P>0.05)。结论年龄增长会导致精液前向运动精子百分率、活动精子百分率、精子顶体酶活性、DFI等参数改变,直接或间接影响男性生育力。说明年龄对男性不育的影响是多方面的,建议有生育需求的大龄(≥40岁)男性尽早进行生育咨询与评估。     

精子形态与精子冷冻复苏率的关系研究

目的 了解精子形态与精子冷冻复苏率之间的关系.方法 135例浙江省人类精子库筛选合格的供精者精液标本,按前向运动精子冷冻复苏率分为冷冻不合格组(<60%)40例和冷冻合格组(≥60%)95例.对所有精液标本进行精子形态学分析.结果 冷冻不合格组正常形态精子百分率(23.53±4.87)%,明显低于冷冻合格组正常形态精子百分率(28.54±544)%,(P<0.01);冷冻不合格组精子头部异常率、畸形精子指数(TZI)及精子畸形指数(SDI)明显高于冷冻合格组(P<0.05);颈部及中段异常率、尾部异常率及胞质小滴两组无明显差异(P>0.05).结论 精子形态对前向运动精子冷冻复苏率有影响,正常形态精子对低温的耐受性好;头部畸形以及同时含有多种畸形的精子对低温冷冻的耐受性差.     

部分型圆头精子症患者精液常规参数及精子形态分析

目的:分析部分型圆头精子症患者精液常规参数及精子形态。方法:选取2013年1月至2016年5月100例部分型圆头精子症患者作为病例组,对照组为非圆头精子症的不育患者180例。根据精液中圆头精子所占比例不同将病例组分成5组:1组(25%~40%)、2组(41%~55%)、3组(56%~70%)、4组(71%~85%)、5组(86%~99%),根据WHO《人类精液检查与处理实验室手册》第5版标准对其进行精液常规分析,包括精子浓度、活动率、前向运动精子百分率,及对不同畸形精子进行计数并计算畸形精子指数(TZI)及精子畸形指数(SDI)。结果:病例组与对照组的精子活动率[(35.76±24.88)%vs(62.03±10.20)%]、前向运动精子百分率[(26.11±20.39)%vs(45.62±6.87)%]、正常形态精子百分率[(1.45±1.45)%vs(5.98±2.21)%]和SDI(1.33±0.11 vs1.27±0.57)相比差异均有统计学意义(P均0.01),两组年龄[(29.82±4.90)岁vs(30.33±3.59)岁]、精子浓度[(46.01±40.38)×10~6/ml vs(54.00±25.15)×10~6/ml]和TZI(1.35±0.11 vs 1.34±0.54)比较差异均无统计学意义(P均0.05);病例组各组精子活动率、前向运动精子百分率、正常形态精子百分率、TZI和SDI均有统计学差异(P均0.01),而年龄和精子浓度均无统计学差异(P均0.05)。随着圆头精子比例的升高,精子头部形态由异质性偏向于均一性。结论:精液中不同比例的圆头精子与精液常规参数和形态学指标密切相关,可对不育患者辅助生殖技术的选择和受精率的预测起一定的提示作用。     

Relationship between the sperm penetration assay and other tests of sperm function

Semen from 88 men of infertile couples and 33 fertile donors differed in seminal fluid analysis (sperm density and motility) (SFA) as well as in the penetration of hamster ova (SPA) and bovine cervical mucus (MPT). In the fertile group, significantly more subjects had adequate SFA, SPA, or MPT results than in the infertile group. When the two groups were subdivided into those with normal or those with abnormal SFA, no differences were noted in SPA, MPT, or postcoital test (PCT) scores. The SFA parameter most consistently reflected in the results of the SPA, MPT, and PCT was sperm density. This was most evident when the SFA was poor. The worst prognosticator of fertility was the SFA, with 30% of the fertile donors having an abnormal SFA. The worst prognosticator of infertility was the MPT, with 79% of the patients penetrating in the fertile range. The SPA was a significantly better predictor than either the SFA or MPT. SPA and MPT results were positively correlated only in the overall infertile group. The SPA, MPT, and PCT measure sperm qualities distinct from those revealed by the SFA, and from each other, and in combination provide the best assessment of fertility.     

精子库捐精志愿者特征与筛查合格率的分析

目的:探讨捐精志愿者年龄、学历、职业、婚育特征与筛查结果的关系,为精子库捐精宣传招募工作提供参考。方法:对广东精子库2003年1月至2017年6月期间12 362例捐精志愿者的筛查资料进行汇总统计,分析捐精志愿者年龄、学历、职业、婚育特征与筛查合格率的关系。结果:12 362例志愿者中,3 968例(32.1%)精液质量达到合格标准;全部检查结果达到精子库捐精标准即筛查合格的志愿者3 127例(25.3%)。志愿者达到精子库捐精标准合格率20~24岁组为27.7%,25~29岁组24.3%,30~34岁组23.8%,≥35岁组17.5%,不同年龄组筛查合格率差异有统计学意义(P0.01)。高中、专科、本科、研究生的志愿者筛查合格率分别为23.5%、24.0%、25.9%和30.3%,差异有统计学意义(P=0.000)。学生和社会人员的筛查合格率分别为29.3%和22.9%,差异有统计学意义(P0.01)。已婚和未婚志愿者的筛查合格率分别为41.5%和20.7%,差异有统计学意义(P0.01)。已生育和未生育志愿者的筛查合格率分别为45.6%和20.9%,差异有统计学意义(P0.01)。结论:人类精子库尽量招募年龄低于35岁、学历大学本科或以上并以学生为主的志愿者,并面向此类人群进行精准宣传,提高他们捐精的积极性和意愿,从而提高精子库筛查合格率。     

Analysis of the outcome of intracytoplasmic sperm injection using fresh or frozen sperm

Study Type – Diagnostic (retrospective cohort)
Level of Evidence 2b What’s known on the subject? and What does the study add? The results of ICSI using fresh or frozen sperm on the site of sperm retrieval remains controversial with respect to outcome. The results of this study showed no difference in outcome using ICSI either with respect to the site of retrieval or whether the sperm used was fresh or frozen. It also showed that the outcome of ICSI is not related to the underlying cause of the azoospermia.

OBJECTIVES

? To compare the outcome of first‐attempt intracytoplasmic sperm injection (ICSI) ICSI–embryo transfer (ET) cycles using frozen‐thawed testicular sperm (FTTS), fresh testicular sperm (FTS), frozen‐thawed epididymal sperm (FTES) and fresh epididymal sperm (FES) so as to determine which of these has the most successful ICSI outcome with respect to fertilization rate (FR), pregnancy rate (PR) and birth rate. ? To assess the outcomes according to the underlying aetiology of azoospermia.

PATIENTS AND METHODS

? The records of 493 patients undergoing first‐attempt ICSI between 1993 and 2008 were reviewed retrospectively. FTS was used in 112 cycles, FTTS in 43 cycles, FES in 279 cycles, and FTES in 59 cycles. ? Within each group, the aetiology of the azoospermia was recorded according to history, clinical examination and histological analysis (n= 316). ? The FR, clinical PR and delivery rate were calculated for each group with respect to the type of sperm retrieval used.

RESULTS

? Analysis of the data showed no significant differences between any of the four groups in the FR, PR or delivery rate (P > 0.05). ? There were no significant differences seen between fresh sperm (FTS and FES) and frozen sperm (FTTS and FTES) or between epididymal sperm (FES and FTES) and testicular sperm (FTS and FTTS) in any of the outcomes measured (P > 0.05). However, sub‐set analysis showed a statistically higher FR and PR for FTTS over fresh sperm. ? When comparing aetiologies, there was no significant difference in the FR, clinical PR and delivery rate between obstructive azoospermia (OA) and non‐obstructive azoospermia (NOA) groups. However, sub‐set analysis showed a higher PR and birth rate for FTTS over fresh sperm in both OA and NOA groups.

CONCLUSIONS

? The results of the present study suggest that using frozen sperm in ICSI cycles is a reliable and favourable method with the same outcome as fresh sperm. ? Testicular and epididymal sperm have similar ICSI outcomes for both fresh and frozen samples. However, results suggest a tendency for higher PRs and birth rates for frozen than for fresh testicular sperm in both OA and NOA aetiologies. ? The aetiology of azoospermia does not significantly affect the outcome of first‐attempt ICSI. The higher rates in the frozen groups suggest that these patients have had better quality semen when they were initially harvested and frozen.     

精子库13723管外供精液标本的妊娠结局分析

目的:探讨使用精子库的精液标本进行辅助生育的安全性及各种助孕方式之间妊娠结局的差异性。方法:对精子库外供给各临床生殖中心的精液标本,采用供精人工授精(AID)、供精体外受精(IVF)及胞质内单精子注射(ICSI)的方式助孕所反馈的资料进行分析比较。结果:精子库共外供13 723管精液标本用于辅助生育,各临床生殖中心使用精液标本的情况不同,有的使用1管/周期,有的使用2管/周期,13 723管精液标本共进行7 743个周期,包括AID 7 123个周期,临床妊娠1 415例(19.87%),其中正常生育1 221例(86.29%),流产数169例(11.94%),出生缺陷6例(0.43%),异位妊娠19例(1.34%),感染性传播疾病0例;IVF 571个周期,临床妊娠367例(64.27%),其中正常生育330例(89.92%),流产数35例(9.54%),出生缺陷0例,异位妊娠2例(0.54%),感染性传播疾病0例;ICSI 49个周期,临床妊娠28例(57.14%),其中正常生育25例(89.29%),流产数3例(10.71%),出生缺陷0例,异位妊娠0例,感染性传播疾病0例。AID与IVF、ICSI的临床妊娠率相比较有显著性差异(P<0.05),而IVF与ICSI的临床妊娠率相比较则无显著性差异(P>0.05)。AID、IVF和ICSI在流产率、出生缺陷率及异位妊娠率方面,三者相比无显著性差异(P>0.05)。结论:无1例因使用精子库的精液标本而感染性传播疾病。采用3种助孕方式的流产率、出生缺陷率和异位妊娠率与通过自然方式受孕之间并无明显差异,因此使用精子库的精液标本进行辅助生育是安全的。IVF和ICSI的妊娠率高于AID,但AID的费用较低,而IVF和ICSI的费用较高,对于究竟采用何种方式助孕,则需根据临床实际情况而定。     

Influence of sperm pretreatment on the efficiency of intracytoplasmic sperm injection in pigs

The purpose of this study was to determine the influence of sperm pretreatment on the efficiency of intracytoplasmic sperm injection (ICSI) in pigs. This was done by examining the effect of 1) the conservation method (fresh vs frozen); 2) the sperm treatment preinjection (resuspension in Dulbecco phosphate-buffered saline (DPBS) vs selection by a Percoll gradient); and 3) the acrosomal and live or dead status of the spermatozoa (by incubation with or without calcium ionophore, 1 muM and 5 muM). In vitro matured porcine oocytes were injected with treated spermatozoa according to each experiment. All the experiments were done with non-artificially activated oocytes. The percentages of activation and cleavage were higher (68% vs 43% and 63% vs 43%, respectively, P < .05) in oocytes injected with fresh vs frozen spermatozoa. The DPBS treatment allowed higher cleavage proportions than the Percoll treatment (P < .05). Moreover, a boar effect was observed in the percentage of developing blastocysts. None of the studied parameters was affected by the acrosomal or the live or dead status of the spermatozoa injected. In conclusion, the use of fresh semen is recommended for porcine ICSI, as well as careful selection of the boar; Percoll treatment is only recommended for poor-quality samples or for removing toxic agents, and no exogenous form of activation or induction of the acrosome reaction is necessary for porcine oocytes to develop a male pronucleus and cleave up to the 2-cell stage after ICSI, although experimental conditions to reach the blastocyst stage need to be investigated further.     

Computerized sperm motility and application of sperm cryopreservation

An objective method for measuring sperm motion characteristics was developed on an Intellect 100 Quantel Image Analysis System suitable for various image cytometric applications. It provided overall analysis of percent motility (% MS) as well as individual and mean measurements of motion characteristics, including vigor characteristics such as curvilinear velocity (Vc), straight line velocity (Vsl), and trajectory pattern characteristics, that is, progressiveness ratio (PR) and amplitude of lateral head displacement (Alh). Evaluation of the method for reproducibility and accuracy showed reliable measurements of these parameters measured on a minimum of 70 motile sperm, sufficient to describe adequately the sperm population. A study was performed comparing motion characteristics of 30 semen samples falling in a normal range before and after cryopreservation in cryoprotector medium (CM). A mean motility rate of recovery (MRR) of 45% was obtained. Only sperm count and concentration in motile forms among initial semen variables correlated weakly with MRR. Velocity recovery rate (VRR) approached 1 with a marked variability among ejaculates. Distribution profile of Vc was highly modified by freezing in CM: spermatozoa that were initially fast and progressive were the most resistant to cryoaggression. PR and Alh values were little affected by freezing in CM. The tolerance of various samples from a given patient was highly variable for % MS and Alh and less variable for Vc and PR. This illustrates the difficulty in predicting the effect of freezing on motility characteristics and, therefore, of extrapolating from semen variables the ability of frozen-thawed samples to fertilize.     

北京地区精子库自精保存现状分析

目的:综合分析北京地区精子库的自精保存现状,更好利用精子库的资源保存男性生育力。方法:对2006年7月至2016年12月在精子库进行自精保存者的基本资料和精液质量进行回顾性分析。结果:251例自精保存者成功保存,2011～2013年自精保存的平均年增长率为119%。自精保存者以本科和硕士学历为主(n=175,69.72%),年龄以20～39岁人群为主(n=204,81.27%),未育者占88.84%(n=223)。有27.49%(n=69)患者冷冻精液数量少于10支,有10.36%(n=26)患者仅冻存1次。最终冻存后精液使用仅为5.58%(n=14)。保存原因中以患肿瘤原因最多(n=140,55.78%),并以血液肿瘤最多(n=66,22.31%),其次为睾丸肿瘤(n=33,13.15%)和其他肿瘤(n=41,16.33%)。肿瘤患者与非肿瘤患者平均精子浓度(60.53×10~6/ml vs 90.45×10~6/ml)、精子总数(175.8×10~6vs 311.3×10~6)、前向运动精子百分率(43.55%vs 49.21%)和前向运动精子的复苏率(52.17%vs 68.13%)有显著性差异(P0.05)。睾丸肿瘤、血液肿瘤及其他肿瘤平均精子浓度分别为37.68×10~6/ml、57.98×10~6/ml、90.69×10~6/ml,精液体积分别为2.73、2.82、3.41 ml,精子总数分别为93.29×10~6、158.41×10~6、349.49×10~6,前向运动精子百分率分别为45.32%、43.47%、44.49%,前向运动精子百分率的复苏率分别为48.32、50.07%、61.09%,睾丸肿瘤患者的精子浓度和总数显著低于另外两组(P0.05)。结论:北京地区自精保存人数呈现逐年增加的趋势,以肿瘤患者最多;多数肿瘤患者错过了最佳保存时期,应提高患者和医务工作者对自精保存的知晓度。     

Cryopreservation of ram semen facilitates sperm DNA damage: relationship between sperm andrological parameters and the sperm chromatin structure assay

We hypothesized that cryopreservation and incubation in conditions that mimic the female genital tract following insemination increases the susceptibility of ram sperm DNA to denaturation. Ram sperm samples (n = 12) underwent the sperm chromatin structure assay (SCSA) and semen quality tests, including motility parameters, viability, and chlortetracycline fluorescence (CTC) patterns. We also assessed correlations between SCSA variables and semen quality parameters. Analyses were performed for both fresh and cryopreserved samples at 0, 3, and 20 hours of incubation in synthetic oviductal fluid (SOF; 39 degrees C, 5% CO(2)). The SCSA variables, mean alpha t (X alpha(t)) and standard deviation of alpha t (SD alpha(t)), were higher because of cryopreservation (P <.05, P <.001, respectively) after 20 hours in SOF. For both fresh and frozen spermatozoa, SCSA values (X alpha(t), SD alpha(t), and the percentage of cells outside the main population of alpha(t) [%COMP alpha(t)]) increased during incubation in SOF. Motility was negatively correlated with both SD alpha(t) and %COMP alpha(t), ranging from -0.39 (P <.01) to -0.59 (P <.001) for both fresh and cryopreserved semen; viability also was negatively correlated with X alpha(t), SD alpha(t), or %COMP alpha(t) (-0.36; P <.05, -.40 and -.46; P <.01, respectively) in fresh semen. The %COMP alpha(t) was positively correlated to the percentage of CTC pattern AR (P <.001) and negatively correlated to the percentages of patterns F and B (-0.33 to -0.60, P <.05 to P <.001). Variation among ejaculates within ram was observed (P <.01). Cryopreservation clearly facilitates DNA damage in physiological conditions. The low to moderate correlations between SCSA variables and classical semen quality parameters indicate that the SCSA provides additional information to standard tests for evaluating ram sperm quality.     

不同来源精子对卵胞浆内单精子注射治疗结局的影响

目的:分析不同来源精子对卵胞浆内单精子注射(ICSI)临床结局的影响.方法:接受ICSI治疗的不育夫妇共进行了286个周期,分为3组,A组(射出精子组):射出精液少弱畸(包括严重少弱畸)精子症186个周期;B组(PESA组):经皮附睾抽吸术(PESA)80个周期;C组(TESE组):睾丸精子获取术(TESE)20个周期,比较其妊娠结局.结果:PESA组受精率明显高于射出精子组和TESE组(88%VS 84%,77%,P<0.01),PESA组的临床妊娠率显著高于射出精子组(47%,33%,P<0.05),射出精子组临床妊娠率高于TESE组,但差异无统计学意义;3组的种植率差异极显著(20.7%,31.4%,13.2%,P<0.01);卵裂率(99%,98%,98%)、流产率(7%,15%,0%)均没有显著差异.结论:PESA来源精子比射出精子组及TESE组的受精率及临床妊娠率有所增高,但TESE和PESA来源精子对临床妊娠率没有影响,射出精子组和TESE组来源精子对临床妊娠率没有影响.对于重度少弱精子症患者和梗阻性无精子症患者,进行ICSI治疗,均有机会获得妊娠.