Non invasive assessment of human oocytes and embryos in assisted reproduction: Review on present practices and future trends

Oocyte and embryo grading is one of the most important steps in assisted reproductive technology to identify the best among cultured embryos for transfer or vitrification. The most commonly used non-invasive method is morphological assessment of gametes and embryos using a microscope. This method despite being simple and cost-effective has interobserver and intraobserver variability and subjectivity and has little to offer about the physiological state of embryos. This review sourced research articles and reviews pertaining to other non-invasive assessment methodologies from Medline and PubMed to collate latest technologies in vogue and identify novel methodologies of the future. The review assesses the current understanding in oocyte and embryo grading and touches upon novel non-invasive techniques and potential biomarkers to identify the best embryo. The latest available literature on time-lapse imaging, hierarchical algorithms, omics (consisting of proteomics and secretomes), miRNAs, mitochondrial RNAs and artificial intelligence has been accessed to summarize the enormous information available, to identify gaps in current interpretations, to identify emerging technologies and to provide direction for future research. This review will greatly benefit anyone practicing assisted reproduction and clinical embryology.     

40岁以上高龄女性体外受精-胚胎移植妊娠结局分析

目的：探讨40岁以上高龄女性体外受精-胚胎移植（IVF-ET）的妊娠结局，旨在为高龄女性提供生育咨询以及为改善高龄女性个体化辅助生殖治疗结局提供临床依据。方法：选择我院生殖中心2015年1月—2017年12月女方年龄≥40岁且使用自身卵子行体外受精的共2 467个治疗周期资料，对各项临床数据进行回顾性分析。结果：40岁及以上行辅助生殖治疗的患者，随着女性年龄增加获卵数明显减少（40～48岁女性平均获卵数分别为2.97、 2.69、2.17、2.01、1.77、1.61、1.68、1.29和1.00，44～48岁与40～43岁依次组间比较均P＜0.05），尤其是44岁以上女性胚胎发育潜能明显降低（40～48岁囊胚形成率分别为48.90%、43.72%、33.67%、34.29%、24.39%、21.14%、26.32%、16.67%和0%，44～48岁与40～43岁组间依次比较均P＜0.05）。共有518个周期行新鲜胚胎移植，结果显示，随女性年龄增加，临床妊娠率（40～48岁临床妊娠率分别为26.92%、21.15%、20.79%、10.96%、18.87%、11.11%、5.88%、0%和0%，43～48岁与40～42岁组间依次比较均P＜0.05）、种植率（40～48岁种植率分别为23.65%、19.51%、17.70%、8.54%、7.49%、10.81%、5.56%、0%和0%，43～48岁与40～42岁组间依次比较均P＜0.05）和活产率均显著降低（40～46岁活产率分别为18.46%、10.58%、9.90%、5.48%、5.66%、2.78%和5.88%，43～46岁与40～42岁组间依次比较均P＜0.05），43岁以上者结局更差。44岁以上女性自然流产率明显增高（40～45岁流产率分别为31.43%、50.00%、52.38%、50.00%、70.00%和75.00%，44～45岁与40～43岁组间依次比较均P＜0.05）。46岁女性仅1例妊娠并分娩，47岁和48岁女性均无成功妊娠。与抗苗勒管激素（AMH）＞1.0 ng/mL组相比，AMH≤1.0 ng/mL组妊娠率、种植率及活产率均显著下降（27.04% vs. 14.74%，22.99% vs. 13.50%，15.88% vs. 7.37%；均P＜0.05），流产率明显升高（41.27% vs. 50.00%，P＜0.05）。结论：≥40岁高龄女性随年龄增长生育力逐渐降低。40～43岁年龄段女性助孕仍有一定的价值，尤其是卵巢仍有一定储备者（AMH＞1.0 ng/mL），但44岁以上女性原则上不再建议ART助孕，对于46岁以上卵巢功能衰竭的女性强烈建议卵子捐赠或收养。     

一种较好的RNA分离制备方法及其在人胚RNA提取中的应用

以钒基核苷酸复合物为核酸酶抑制剂,从人胚组织中制备总RNA,所得产品几乎不含DNA和蛋白质,经过分离Poly(A)~ mRNA,证明制品有合成cDNA的完整功能,方法比较简捷,适于大量制备以满足分离mRNA 之需,并讨论了此法的优缺点。     

输卵管积液影响体外培养胚胎质量的实验研究

目的：分析输卵管积液的形成及其化学成分，探讨输卵管积液对体外培养胚胎生长发育的影响及其机制。方法：建立输卵管积液动物模型，测定其成分；用不同浓度输卵管积液培养小鼠2-细胞胚胎，观察胚胎形态。结果：输卵管积认中K^ 、HCO3^-和表皮生长因子（EGF）含量明显低于输卵管液；pH值明显增高。输卵管积液组囊胚形成率和胚胎孵化率均明显低于输卵管液。结论：结扎兔输管两端能诱导输卵管积液形成；机械诱导的输卵管积液成分改变能影响早期胚胎的质量。     

Determination of diffusion coefficient of hydrogen in metals and their elastic modulus by generalised impedance

Transport of hydrogen in elastic metals has been analysed in terms of an impedance-like non-linear transfer function (TF). The equilibrium in a membrane specimen is perturbed by a sinusoidal signal of the concentration of hydrogen, of large amplitude, applied to one of its surfaces. The fundamental harmonics of periodically reproducible flux at the same surface is considered as a response. The TF is defined as the ratio of the signal to the response. The power of the said TF in determining the diffusion coefficient of hydrogen in a metal–hydrogen system and its bulk elastic modulus was analysed for the example of α-phase Pd–H and Pd₈₁Pt₁₉–H systems. It has been found that such a TF is suitable for determination of the above parameters. Also, the sensitivity of these estimates to the uncertainty of concentration of hydrogen in the metal, its partial molar volume and the thickness of the specimen has been explored.     

预移植在胚胎移植中的意义

目的 :评价预移植在胚胎移植中的作用。　方法 :在超声波引导下胚胎移植 114个周期 ,其中 10 1个周期进行预移植。比较预移植与未预移植周期临床妊娠率和胚胎种植率 ;按预移植与实际胚胎放置深度差值分组 ,并比较两组妊娠率和胚胎种植率。　结果 :预移植周期与未预移植周期临床妊娠率和种植率统计学差异无显著性(5 9.4 1%和 6 1.5 4 % ,35 .81%和 31.4 3% ,P均 >0 .0 5 ) ;按预移植与实际胚胎放置深度差值分组后 ,两组妊娠率和种植率统计学差异也无显著性 (6 1.5 4 %和 5 8.6 7% ,4 0 .79%和 34.0 9% ,P均 >0 .0 5 )。　结论 :在胚胎移植技术中 ,预移植不能准确指引胚胎移植深度     

Ballistocardiogram of avian eggs determined by an electromagnetic induction coil

As an avian embryo grows within a eggshell, the whole egg is moved by embryonic activity and also by the embryonic heartbeat. A technical interest in detecting minute biological movements has prompted the development of techniques and systems to measure the cardiogenic ballistic movement of the egg or ballistocardiogram. (BCG). In this context, there is interest in using an electromagnetic induction coil (solenoid) as another simple sensor to measure the BCG and examining its possibility for BCG measurement. A small permanent magnet is attached tightly to the surface of an incubated egg, and then the egg with the magnet is placed in a solenoid. Preliminary model analysis is made to design a setup of the egg, magnet and solenoid coupling system. Then, simultaneous measurement with a laser displacement measuring system, developed previously, is made for chicken eggs, indicating that the solenoid detects the minute cardiogenic ballistic movements and that the BCG determined is a measure of the velocity of egg movements.     

低剂量阿司匹林在辅助生育技术中的应用

目的　探讨在进行体外受精 -胚胎移植 ( IVF-ET)或卵母细胞浆内单精子注射 ( ICSI)患者口服低剂量阿司匹林对治疗结果的影响。　方法　 IVF或 ICSI治疗的 3 2 4个 ET周期 ,分为是否服用阿司匹林两组 ,分别统计患者注射人绒毛膜促性腺激素 ( h CG)时子宫内膜厚度及 ET胚胎数 ,比较两组的胚胎种植率、临床妊娠率及流产率 ,并进行统计学处理。　结果　两组在注射 h CG日内膜平均厚度、ET中优质胚胎的比率及流产率均无显著性差异 ;未服用阿司匹林组的 ET胚胎数明显多于服用组 ( P<0 .0 1 ) ,而前者的种植率、临床妊娠率却明显低于后者 ( P<0 .0 1 )。　结论　低剂量阿司匹林在 IVF-ET或ICSI的辅助生殖治疗中能有效地提高胚胎种植率 ,增加患者的临床妊娠率     

流产鼠Caspase-3与laminin B和PAI-1的表达及对妊娠结局的影响

目的:研究阻断CD86协同刺激分子对自然流产模型孕鼠母胎界面Caspase-3、laminin B和PAI-1的表达及对妊娠结局的影响。方法:实验组于妊娠第4.5天腹腔注射大鼠抗小鼠CD86单抗,实验对照组注射大鼠同型IgG2b,正常妊娠组不作任何处理。于妊娠第13.5天计算胚胎吸收率,用免疫组化测定Caspase-3、laminin B和PAI-1的表达,并进行图像分析、检测免疫组化染色灰度值(A)。结果:①实验组的胚胎吸收率显著低于实验对照组(P<0.05);②实验组中Caspase-3蛋白灰度值明显高于实验对照组(P<0.05),实验组中laminin B和PAI-1蛋白灰度值均明显低于实验对照组(P<0.05)。结论:妊娠早期阻断CD86协同刺激分子可使母胎界面中的Caspase-3、laminin B和PAI-1分别通过各自不同的途径发挥免疫耐受作用并且使自然流产模型孕鼠的胚胎吸收率降低至正常妊娠水平。     

达菲林在体外受精-胚胎移植中的应用

目的:观察达菲林在体外受精-胚胎移植中的疗效。方法:根据112例不孕患者用药前窦卵泡的数目采用不同剂量的达菲林降调节,常规超促排卵至卵泡成熟后,经阴道B超引导下取卵,行实验室体外培养受精,优质胚胎移植。结果:3组获卵数、获得的Ⅰ级胚胎数、胚胎移植后临床妊娠率、卵巢过度刺激综合征发生率等均无明显差异,受精率皆在70％以上,卵裂率在93％以上。结论:不同剂量的达菲林降调节均可获得高质量的卵子,且是一种肌肉注射缓释剂,易被患者接受。