授精方式对不同原核数目及D3评级胚胎继续发育能力的影响

目的评估授精方式对不同原核数目及D3评级胚胎体外发育形成囊胚能力的影响。方法回顾性分析2015年4月至2017年2月期间深圳中山泌尿外科医院生殖医学中心3 610个新鲜取卵周期的囊胚培养结局。根据授精方式将行囊胚培养的胚胎分为IVF组和ICSI组,每组再根据原核数目(0PN、1PN和2PN)以及受精后第3天(D3)卵裂期胚胎评级(优质和非优质,分别简称为H及L)将胚胎分为6个亚组,即2PN-H、2PN-L、1PN-H、1PN-L、0PN-H以及0PN-L亚组,比较分析各组的囊胚形成率(BR)、优质囊胚形成率(HBR)、可利用囊胚形成率(ABR)、D5BR、D5 HBR、D5ABR等指标。结果 ICSI组的总BR、HBR以及ABR与IVF组相比均无显著性差异(P0.05)。ICSI-1PN-H亚组、ICSI-1PN-L亚组的BR、HBR、ABR以及ICSI-0PN-L亚组的BR、ABR均显著性地低于IVF相应亚组(P0.05),而ICSI-2PN-H亚组、ICSI-2PNL亚组以及ICSI-0PN-H亚组的BR、HBR以及ABR与IVF相应亚组间的差异无统计学意义(P0.05)(除ICSI-2PN-L亚组的ABR外)。另外,ICSI组的总D5BR、D5 HBR以及D5ABR均显著低于IVF组(P0.01),但是ICSI-0PN-H亚组的D5BR、D5HBR、D5ABR却显著高于IVF-0PN-H亚组(P0.01)。结论通过ICSI获得的1PN胚胎(无论D3胚胎评级)以及0PN-L胚胎的继续发育能力较低,并且ICSI会降低大部分胚胎的发育速度。     

维生素D对复发性流产患者外周血自然杀伤细胞的调节作用

目的探讨维生素D对复发性流产(recurrent miscarriage,RM)患者外周血自然杀伤细胞(natural killer cells,NK)的调节作用。方法收集99例RM患者黄体中期的外周血,流式细胞术检测外周血NK细胞的数量和细胞毒性。根据患者体内维生素D水平将其分为三组:维生素D正常组(vitamin D normal,VDN)、维生素D不足组(vitamin D insufficiency,VDI)和维生素D缺乏组(vitamin D deficiency,VDD)。对维生素D缺乏和不足的RM患者补充1,25(OH)2D,分析维生素D补充对RM患者的NK细胞的调节作用。结果 CD3~-CD56~+NK细胞比例在三组间无显著差异(P0.05)。VDI组中NK细胞毒性在效靶比(effector-to-target ratio,E:T)为50∶1(51.6±8.8 vs.45.3±12.4)(P0.05)、25∶1(43.2±9.1 vs.35.8±11.8)(P0.01)和12.5∶1(30.2±10.3 vs.23.0±10.6)(P0.01)这三个条件下均显著高于VDN组。补充维生素D后,可显著降低E:T为50∶1(41.6±14.0 vs.51.0±10.5)(P0.01)、25∶1(34.9±14.3 vs.43.1±10.8)(P0.01)和12.5∶1(22.6±11.4 vs.29.9±10.7)(P0.01)的NK细胞毒性。结论低水平维生素D的RM患者外周血NK细胞毒性增加,而补充1,25(OH)2D可在一定程度上降调NK细胞毒性。     

不明原因复发性自然流产患者分泌期子宫内膜树突状细胞数量分析

目的研究子宫内膜树突状细胞(DC)在不明原因复发性自然流产(uRSA)患者分泌期子宫内膜的表达情况。方法根据子宫内膜形态学Noyes标准,对在本中心就诊的uRSA患者(uRSA组,n=113)与男性因素不孕患者(对照组,n=35)进行子宫内膜分期;以CD1_a与CD83分别作为子宫内膜未成熟树突状细胞(iDC)与成熟树突状细胞(m DC)的表面标志物,并利用免疫组织化学技术检测患者分泌期子宫内膜CD1_a与CD83阳性细胞的数量,并通过Vectra~?自动病理成像定量分析系统分别计算其阳性细胞率。结果 uRSA组分泌期子宫内膜CD1_a阳性细胞率无明显变化,而CD83阳性细胞率[(2.03±1.60)%]显著高于对照组[(0.81±1.11)%,P=0.000];两组患者子宫内膜CD1_a阳性细胞率从分泌早期到中期均呈递减趋势,而CD83阳性细胞率从分泌早期到中期均呈递增趋势;与对照组相比,uRSA组子宫内膜CD1_a阳性细胞率在分泌早期和中期均无明显变化,而uRSA组子宫内膜CD83阳性细胞率在分泌早期[(1.64±1.20)%]和中期[(2.20±1.60)%]均显著高于对照组[(0.17±0.11)%,P=0.000;(0.63±0.78)%,P=0.000]。结论 uRSA组患者子宫内膜分泌早期和中期CD83阳性细胞率异常升高,提示其可能影响子宫内膜容受性,从而导致妊娠失败。     

重组人黄体生成素在体外受精-胚胎移植促排卵治疗中的应用

Objective To evaluate application of recombinant human luteinizing hormone (r-hLH)used in ovarian stimulation of assisted reproductive technique and impact on outcome of pregnancy. Methods From Apr. To Jul. 2009, 123 patients with low LH level ( < 1 U/L) at day 3 of menstruation and downregulation of pituitary function undergoing in vitro fertilization-embryo transfer (IVF-ET) in Reproductive Medical Center, Provincial Hospital Affiliated to Shandong University were enrolled in this study, whom were classified into 66 cases treated by r-hLH in r-hLH group and 57 cases without r-hLH treatment in non-r-hLH group. In the mean time, 145 patients with normal level of serum LH ( 1-2 U/L) not given by r-hLH treatment and undergoing IVF-ET were matched as control group. Total amount of gonadotropin, estradiol levels and LH levels on the administration of human chorionic gonadotropin ( hCG), number of oocytes retrieved, number of 2PN zygotes, rate of high quality embryos, the rates of implantation and clinical pregnancy were compared among these three groups. Results The level of serum LH on the day of hCG administration were ( 1.59 ± 0.77 ) U/L in r-hLH group, (0.54 ± 0.25 ) U/L in non-r-hLH group and (2.39 ± 1.01 ) U/L in control group, which reached tatistical difference between every two groups (P < 0.05). The rates of high quality embryo were 59.36％ in r-hLH group, 57.79％ in non-r-hLH group,which were significantly lower than 65.94％ in control group, respectively (P < 0. 05 ). The rates of 2PN were 67.62％ in r-hLH group and 68. 32％ in control group, which were significantly higher than 62. 84％ in non-r-hLH group, respectively ( P < 0.05 ). The rates of implantation of 29.77％ in r-hLH group were significantly higher than 18.26％ in non-r-hLH group ( P < 0.05 ). The total amount of gonadotropin,estradiol level on the day of hCG administration, the number of oocytes retrieved, and clinical pregnancy rate were not significantly different among those three groups ( P > 0.05 ). Conclusion The administration of recombinant human uteinizing hormone in patients who are profoundly suppressed after down-regulation with long protocol can get more quality embryos, the higher rates of 2PN and implantation.     

GnRH-a扳机对PCOS患者新鲜胚胎移植临床妊娠率的影响

目的研究促性腺激素释放激素激动剂(GnRH-a)扳机对多囊卵巢综合征(PCOS)患者新鲜周期移植临床妊娠率的影响。方法回顾性分析2016年1月1日至2018年6月30日就诊本院生殖中心行IVF/ICSI的442例PCOS患者的临床资料,根据扳机日使用的扳机方案分为两组:GnRH-a扳机组(163例),在扳机日使用GnRH-a 0.2mg扳机;HCG扳机组(279例),在扳机日使用HCG 10 000U扳机。卵巢过度刺激综合征(OHSS)低风险者(GnRH-a扳机组68例,HCG扳机组79例)取卵后5d移植1枚囊胚。GnRH-a扳机组黄体支持从取卵当天开始使用戊酸雌二醇(4mg/d)、地屈孕酮(40mg/d)和黄体酮阴道缓释凝胶(90mg/d),至孕10周停药。HCG扳机组黄体支持不使用戊酸雌二醇,其余同GnRH-a扳机组。观察获卵数、可利用胚胎数、MⅡ卵率、鲜胚移植周期临床妊娠率、持续妊娠率、早期流产率及OHSS发生率。结果两组之间的获卵数[(23.03±7.95)vs.(21.92±8.43)]、MⅡ卵率(89.60%vs.88.90%)、可利用胚胎数[(8.93±5.61)vs.(8.80±4.95)]、临床妊娠率(55.88%vs.65.82%)、持续妊娠率(48.52%vs.56.96%)及早期流产率(13.16%vs.18.42%)差异均无统计学意义(P0.05)。HCG扳机组的OHSS发生率高于GnRH-a扳机组(8.24%vs.1.23%),差异有统计学意义(P0.05)。结论本研究中GnRH-a扳机未降低PCOS患者新鲜周期移植的临床妊娠率,但显著降低OHSS的发生率。     

胚胎培养微滴内的温度变化研究

目的研究胚胎培养皿(以下简称培养皿)从胚胎培养箱取出后放置于恒温热平板(以下简称热平板)上时胚胎培养微滴(以下简称微滴)内的温度变化及放回4种不同类型胚胎培养箱后的复温时间。方法本研究选择4种不同类型的商品化培养箱,#1为大型箱式培养箱;#2为小型箱式培养箱;#3为桌面干式培养箱;#4为桌面湿式培养箱。将商品化的温度测定仪温度探头偶联在胚胎培养皿上,自组装一套简易装置实现对培养皿内微滴温度检测。将胚胎培养皿从CO2培养箱取出后放置在热平板上,采用该装置每30s测量一次培养皿中微滴内的温度的变化,温度下降至30℃时分别将培养皿放回4种不同培养箱中,记录温度恢复至37℃的复温时间。结果胚胎培养皿离开CO2培养箱,随着在热平板上操作时间的延长,微滴内的温度逐渐降低。离箱操作3 min,微滴温度降至(36.50±0.52)℃;离箱操作5 min,微滴温度降至(36.17±0.67)℃;10min时降至(35.53±0.64)℃;10min以后,微滴温度趋于平稳。培养皿在重新放回培养箱后,35℃恢复至37℃的时间分别为(5.58±1.37)min、(15.69±5.19)min、(5.21±0.45)min、(4.88±0.51)min,其中#2培养箱的复温时间显著长于#1、#3、#4培养箱(P0.05),#4培养箱的复温时间最短,但#1、#3和#4培养箱之间的复温时间无显著性差异(P0.05)。结论从温度上考虑,结合已有的报道,建议胚胎离开CO2培养箱在热平板的时间应控制在3min内。在重新放回胚胎培养箱后,不同类型培养箱内微滴的复温时间是有差异的,在实际工作中,应尽量选择复温时间短的培养箱以优化胚胎培养效果。     

黄体生成素在辅助生殖技术促排卵周期中的应用

在女性的排卵周期中,卵泡的正常发育是由垂体分泌的卵泡刺激素(follicle stimulating hormone,FSH)和黄体生成素(luteinizing hormone,LH)两种激素控制的,两者对于正常卵泡雌二醇的生物合成都很重要,也就是所谓的"两种细胞,两种促性腺激素"学说,即FSH刺激卵泡发育,LH与卵泡膜细胞上的受体结合,激发雄激素前体从膜细胞转移至颗粒细胞,并通过FSH刺激芳香酶将雄激素转变为雌激素.     

重组人黄体生成素在体外受精-胚胎移植促排卵治疗中的应用

Objective To evaluate application of recombinant human luteinizing hormone (r-hLH)used in ovarian stimulation of assisted reproductive technique and impact on outcome of pregnancy. Methods From Apr. To Jul. 2009, 123 patients with low LH level ( < 1 U/L) at day 3 of menstruation and downregulation of pituitary function undergoing in vitro fertilization-embryo transfer (IVF-ET) in Reproductive Medical Center, Provincial Hospital Affiliated to Shandong University were enrolled in this study, whom were classified into 66 cases treated by r-hLH in r-hLH group and 57 cases without r-hLH treatment in non-r-hLH group. In the mean time, 145 patients with normal level of serum LH ( 1-2 U/L) not given by r-hLH treatment and undergoing IVF-ET were matched as control group. Total amount of gonadotropin, estradiol levels and LH levels on the administration of human chorionic gonadotropin ( hCG), number of oocytes retrieved, number of 2PN zygotes, rate of high quality embryos, the rates of implantation and clinical pregnancy were compared among these three groups. Results The level of serum LH on the day of hCG administration were ( 1.59 ± 0.77 ) U/L in r-hLH group, (0.54 ± 0.25 ) U/L in non-r-hLH group and (2.39 ± 1.01 ) U/L in control group, which reached tatistical difference between every two groups (P < 0.05). The rates of high quality embryo were 59.36％ in r-hLH group, 57.79％ in non-r-hLH group,which were significantly lower than 65.94％ in control group, respectively (P < 0. 05 ). The rates of 2PN were 67.62％ in r-hLH group and 68. 32％ in control group, which were significantly higher than 62. 84％ in non-r-hLH group, respectively ( P < 0.05 ). The rates of implantation of 29.77％ in r-hLH group were significantly higher than 18.26％ in non-r-hLH group ( P < 0.05 ). The total amount of gonadotropin,estradiol level on the day of hCG administration, the number of oocytes retrieved, and clinical pregnancy rate were not significantly different among those three groups ( P > 0.05 ). Conclusion The administration of recombinant human uteinizing hormone in patients who are profoundly suppressed after down-regulation with long protocol can get more quality embryos, the higher rates of 2PN and implantation.     

黄体期促排卵在卵巢低反应患者中的应用

目的探讨体外受精-胚胎移植(IVF-ET)周期中,黄体期促排卵在卵巢低反应(POR)患者中的应用及临床体会。方法回顾性分析POR的227个IVF周期,分析克罗米芬微刺激阶段和后续的黄体期促排卵阶段的一般资料、获卵数、取消率、再获卵率等及冷冻胚胎移植妊娠结局等指标。总结黄体期促排卵应用于POR患者中的临床体会。结果两个阶段间MII卵率、受精率、优质胚胎率均无统计学差异(P0.05),但黄体期促排卵阶段的平均获卵数明显高于微刺激阶段(P0.05)。微刺激阶段及黄体期促排卵阶段所获得的冷冻胚胎进行移植后,临床妊娠率为28.8%,胚胎种植率为15%。黄体期促排卵方案的再获卵率64.4%。结论在POR患者中,黄体期促排卵方案周期获卵数及再获卵率高,能降低患者心理压力,获得较满意的临床妊娠结局,提示黄体期促排卵方案对于POR患者是一种值得尝试的促排卵方案。