配子/胚胎操作和管理的安全性

辅助生殖技术（ART）的特点是通过对配子、胚胎的体外操作获得受精卵和早期胚胎,并将胚胎移植入子宫,以解决不孕不育夫妇的生育问题,对配子/胚胎操作和管理的安全性问题显得尤为重要。从配子的获取至胚胎的体外培养、冷冻保存和胚胎移植,是多步骤、多操作的工作流程,潜在着巨大的误差风险,牵涉到社会伦理问题,因此需建立明确的操作流程和管理制度将潜在风险最小化。初步探讨如何提高在辅助生殖技术流程中对配子/胚胎操作和管理的安全性,以最大程度地保证患者的利益,并使中国辅助生殖技术能够在符合社会伦理道德范畴内获得长足发展。     

辅助生殖技术和生殖健康关系的思考

辅助生殖技术的迅猛发展引发了对其所致生殖健康不良影响的思考。应用于辅助生殖技术的超促排卵本身是否对妊娠存在不良影响,超促排卵是否增加卵巢恶性疾病的发生率;各项辅助生殖技术中所包含的配子和胚胎体外操作,配子和胚胎冻融以及多胎妊娠等是否对子代健康存在不良影响,都是近年来探讨的热点问题。另外,辅助生殖技术中涉及的环境物理,化学因素以及心理因素与生殖健康的问题也逐渐受到重视。仅就辅助生殖技术影响生殖健康的环节以及潜在的机制作一述评,以有效评估目前辅助生殖技术的安全性,对辅助生殖临床具有一定的借鉴作用。     

辅助生殖技术和生殖健康关系的思考

辅助生殖技术的迅猛发展引发了对其所致生殖健康不良影响的思考.应用于辅助生殖技术的超促排卵本身是否对妊娠存在不良影响,超促排卵是否增加卵巢恶性疾病的发生率;各项辅助生殖技术中所包含的配子和胚胎体外操作.配子和胚胎冻融以及多胎妊娠等是否对子代健康存在不良影响,都是近年来探讨的热点问题.另外,辅助生殖技术中涉及的环境物理,化学因素以及心理因素与生殖健康的问题也逐渐受到重视.仅就辅助生殖技术影响生殖健康的环节以及潜在的机制作一述评,以有效评估目前辅助生殖技术的安全性,对辅助生殖临床具有一定的借鉴作用.     

人类辅助生殖技术研究进展

自从世界上第1例"试管婴儿"诞生以来,人类辅助生殖技术已经走过了30年的发展历程。随着人们对配子与胚胎培养条件的不断优化,辅助生殖技术的成功率显著提高,现已成为治疗不孕不育症的理想治疗措施之一。近年来,一系列辅助生殖相关技术,如卵子体外成熟、移植前遗传学诊断、单胚胎移植以及冷冻技术等获得了较大的发展,为降低常规体外受精—胚胎移植的费用、风险等提供了可能。该文主要就当前临床及实验性辅助生殖技术的相关研究进展作以综述。     

卵母细胞基因组印迹与辅助生殖技术

基因组印迹控制基因的转录与生殖及子代的健康发育密切相关。在原始生殖细胞阶段基因组印迹擦除,而配子的形成阶段印迹重新建立。辅助生殖技术将配子和早期胚胎暴露于体外并进行不同程度的操作,使辅助生殖技术子代印迹基因疾病风险增加。研究印迹基因疾病发病机制及与体外受精的关系,对正确评价该技术并提高辅助生殖子代健康有重要意义。     

卵母细胞基因组印迹与辅助生殖技术

基因组印迹控制基因的转录与生殖及子代的健康发育密切相关.在原始生殖细胞阶段基因组印迹擦除,而配子的形成阶段印迹重新建立.辅助生殖技术将配子和早期胚胎暴露于体外并进行不同程度的操作,使辅助生殖技术子代印迹基因疾病风险增加.研究印迹基因疾病发病机制及与体外受精的关系,对正确评价该技术并提高辅助生殖子代健康有重要意义.     

辅助生育技术的并发症

人类辅助生育技术就像一把双刃剑,在为不孕夫妇带来福音,造福千千万万个家庭的同时,也可能会出现一系列并发症.因为辅助生殖技术与自然生育过程相比较,存在一些非自然的操作,如促排卵、对卵子或精子的操作、配子在体外环境下受精、胚胎的体外培养、对胚胎的操作、胚胎移植等,每个实施步骤都可能或者影响妊娠的胚胎数目、部位,或者因为技术过程带来的相应问题.下面就简单谈谈实施辅助生育技术可能引发的并发症,希望引起大家注意.     

拉曼光谱检测在辅助生殖领域的研究进展

辅助生殖技术（ART）的体外受精/胞浆内单精子注射-胚胎移植（IVF/ICSI-ET）需要实验室条件下处理两种配子、体外受精、体外培养形成胚胎或囊胚并挑选质量好的胚胎进行移植,使母亲妊娠,是目前解决不孕不育最有效的手段。因此,客观评估配子及植入前胚胎的质量显得非常重要。拉曼光谱不受被测物形态限制,无需样本制备或标记等复杂操作,属于无损伤检测。由于其具有检测迅速、耗时短、敏感度高及分析准确等特点,因此近年被广泛应用于快速检测蛋白质、核酸和脂类等生物大分子损伤。拉曼光谱技术主要有表面增强拉曼光谱技术、共焦显微拉曼光谱技术和傅里叶变换拉曼光谱技术3种。本文概述拉曼光谱的特点及上述3种主要拉曼光谱技术在生物样品分析方面的应用,探讨拉曼光谱技术应用于对精液、卵子和植入前胚胎质量检测的可能性,在辅助生殖领域具有潜在的应用前景。     

拉曼光谱检测在辅助生殖领域的研究进展

辅助生殖技术(ART)的体外受精/胞浆内单精子注射-胚胎移植(IVF/ICSI-ET)需要实验室条件下处理两种配子、体外受精、体外培养形成胚胎或囊胚并挑选质量好的胚胎进行移植,使母亲妊娠,是目前解决不孕不育最有效的手段。因此,客观评估配子及植入前胚胎的质量显得非常重要。拉曼光谱不受被测物形态限制,无需样本制备或标记等复杂操作,属于无损伤检测。由于其具有检测迅速、耗时短、敏感度高及分析准确等特点,因此近年被广泛应用于快速检测蛋白质、核酸和脂类等生物大分子损伤。拉曼光谱技术主要有表面增强拉曼光谱技术、共焦显微拉曼光谱技术和傅里叶变换拉曼光谱技术3种。本文概述拉曼光谱的特点及上述3种主要拉曼光谱技术在生物样品分析方面的应用,探讨拉曼光谱技术应用于对精液、卵子和植入前胚胎质量检测的可能性,在辅助生殖领域具有潜在的应用前景。     

辅助生殖技术对母亲和子代代谢影响的研究进展

<正>目前,不孕已成为全球性的健康问题,影响了约20%的孕龄期夫妻^[1]。辅助生殖技术(assisted reproductive technology,ART)是治疗不孕不育的重要方法。ART由多种操作构成,包括卵巢刺激、配子与胚胎的操作、冷冻保存、体外培养、胚胎移植等^[2],不同的操作干预程度不同,如体外受精(in vitro fertilization,IVF)干预较小,胞浆内精子注射(intracytoplasmic sperm injection,ICSI)和胚胎植入前遗传学检测(preimplantation genetic testing,PGT)的干预较大。     

感染性疾病患者实施辅助生殖技术的风险与防范

在辅助生殖技术（ART）中,临床改善内分泌措施改变了女性生殖道的微生物防御机制,体外对配子/胚胎操作和移植替代了性交,而临床操作对于女性是个风险,有可能导致病原微生物的机械传播,同时也可能被用于患性传播疾病（STD）的不育男性。应高度关注在ART实施过程中女性生殖道的微生物状况和感染问题,避免人为机械性地将病原生物植入女性盆腔深部组织。应用ART避免男性STD传播问题应当慎重。在这种情况下,安全的精子分离程序、规范的实验室操作、患者充分知情同意是必须的。对ART中传播人类免疫缺陷病毒（HIV）的风险应当从生物学、操作程序和伦理学方面再评价。规范的实验室管理是避免因实验室环节疏忽导致病原污染事件发生的必要手段。     

Ex vivo early embryo development and effects on gene expression and imprinting

The environment to which the mammalian embryo is exposed during the preimplantation period of development has a profound effect on the physiology and viability of the conceptus. It has been demonstrated that conditions that alter gene expression, and in some instances the imprinting status of specific genes, have all previously been shown to adversely affect cell physiology. Thus, questions are raised regarding the aetiology of abnormal gene expression and altered imprinting patterns, and whether problems can be averted by using more physiological culture conditions. It is also of note that the sensitivity of the embryo to its surroundings decreases as development proceeds. Post compaction, environmental conditions have a lesser effect on gene function. This, therefore, has implications regarding the conditions used for IVF and the culture of the cleavage stage embryo. The developmental competence of the oocyte also impacts gene expression in the embryo, and therefore superovulation has been implicated in abnormal methylation and imprinting in the resultant embryo. Furthermore, the genetics and dietary status of the mother have a profound impact on embryo development and gene expression. The significance of specific animal models for human assisted reproductive technologies (ART) is questioned, given that most cattle data have been obtained from in vitro-matured oocytes and that genes imprinted in domestic and laboratory animals are not necessarily imprinted in the human. Patients treated with ART have fertility problems, which in turn may predispose their gametes or embryos to greater sensitivities to the process of ART. Whether this is from the drugs involved in the ovulation induction or from the IVF, intracytoplasmic sperm injection or culture procedures themselves remains to be determined. Alternatively, it may be that epigenetic alterations are associated with infertility and symptoms are subsequently revealed through ART. Whatever the aetiology, continued long-term monitoring of the children conceived through ART is warranted.     

乙型肝炎患者实施辅助生殖技术中病毒传播的风险与防范

慢性乙型肝炎（简称乙肝）是中国第一大传染病,严重危害健康和威胁生命,受到广泛关注。随着乙肝患者发病率的上升及辅助生殖技术（ART）的迅猛发展,乙型肝炎病毒（HBV）感染者实施ART过程中HBV的传播成为热议话题。对慢性乙肝患者实施ART,因操作不当及管理不严,有发生交叉感染或院内感染的危险,既可能将病毒传播给正常者的配子和胚胎,也可能将病毒传播给医疗技术人员。然而,科学合理地ART可降低病毒传播给子代的风险。综述HBV感染者的ART传播风险和防范措施,为预防病毒传播和医源性感染提供科学依据。     

The role of animal studies in supporting human assisted reproductive technology

Although average success rates of human IVF have increased progressively during the past two decades, the efficiency of this technique, based on each embryo produced or transferred, is still low. High success rates are usually achieved by transferring several embryos to the patient, which is often associated with multiple pregnancies. The quality of in vitro produced embryos is a major area that needs attention. Because there is no in vivo database for human embryos, the properties of normal embryos are not known, and so it is difficult to know how to improve quality and viability. In addition, selection of the most viable embryos for transfer is a rather subjective process. The origins of human assisted reproductive technology (ART) are based on animal ART; however, the two areas of research (animal and human ART) appear to have become disconnected. Re-examination of progress in animal ART could help improve human embryo quality and thereby assist efforts to sustain high pregnancy rates with only one or two embryos transferred. Some key areas in which animal ART can help guide progress in human ART are discussed.     

Computational modelling of maternal interactions with spermatozoa: potentials and prospects

Understanding the complex interactions between gametes, embryos and the maternal tract is required knowledge for combating infertility and developing new methods of contraception. Here we present some main aspects of spermatozoa interactions with the mammalian oviduct before fertilisation and discuss how computational modelling can be used as an invaluable aid to experimental investigation in this field. A complete predictive computational model of gamete and embryo interactions with the female reproductive tract is a long way off. However, the enormity of this task should not discourage us from working towards it. Computational modelling allows us to investigate aspects of maternal communication with gametes and embryos, which are financially, ethically or practically difficult to look at experimentally. In silico models of maternal communication with gametes and embryos can be used as tools to complement in vivo experiments, in the same way as in vitro and in situ models.     

Clinician-induced (iatrogenic) damage incurred during human infertility treatment: Detrimental effects of sperm selection methods and cryopreservation upon the viability,DNA integrity,and function of human sperm

Human infertility is becoming increasingly prevalent and is now estimated to affect 10% of couples worldwide. Although phenomenal development in assisted reproductive technology (ART) has yielded a powerful and sophisticated discipline with which to combat many types of human infertility, success rates vary markedly and rarely exceed 40%. Mounting evidence suggests that laboratory techniques used routinely in ART may inadvertently impart detrimental effects upon gamete and embryo viability and competence. In this mini-review, we discuss how routine cryopreservation methods, commonly used in ART, may exert iatrogenic (clinician-induced) damage upon sperm structure, DNA integrity, and function. It is recommended that future research programmes aim to refine or replace current cryobiology protocols in order that the efficacy of ART can be optimised accordingly.     

Emergency prevention of extinction of a transgenic allele in a less-fertile transgenic mouse line by crossing with an inbred or outbred mouse strain coupled with assisted reproductive technologies

Certain transgenic mouse lines are difficult to breed or archive and, consequently, their transgenes become lost. We examined a C57BL/6 mouse line (B6-tg), transgenic for green fluorescent protein (GFP) with low fertility, and its crosses with the more prolific inbred C3HeB/FeJ (C3) and outbred Swiss (SW) strains in order to assess the possibility of emergency prevention of extinction of a transgenic allele by using assisted reproductive technologies (ART). Out-crossing was performed by natural mating or in vitro fertilisation (IVF) with heterozygous mice. Most of the crossing combinations resulted in improved archiving and rederivation efficiencies of the transgenic allele. Natural crossing increased both mean litter size by two to three pups and the superovulatory rate from 69% for B6-tg to 70-90% for females from the out-crosses. Each plug-positive B6-tg female yielded an average of 4.6 two-cell embryos, whereas females from the out-crosses produced three- to fivefold that amount. After thawing, 13% of B6-tg embryos and 6-12% of out-crossed embryos developed into transgenic pups after transfer into recipients. After IVF with cryopreserved spermatozoa, cleavage rates were 4% for B6-tg, 22-37% for B6-tg oocytes out-crossed with C3 and SW spermatozoa, 9-49% for gametes from out-crossed mice and 28-44% for back-crosses with B6 oocytes. Transgenic pups were not derived from IVF with B6-tg gametes when either fresh or cryopreserved spermatozoa were used. Rederivation efficiencies were 7% and 4% from out-crosses of B6-tg oocytes with C3 and SW spermatozoa, respectively, 6-22% for gametes from out-crossed mice and 4-10% for the back-crosses. Although out-crossing changes the original genetic background, the strategy of crossing coupled with ART prevents the extinction of an allele of interest, especially where archiving and rederivation of the transgenic line fail.     

颗粒细胞与卵母细胞关系研究进展

颗粒细胞与卵母细胞共处于同一个微环境中,颗粒细胞通过复杂的缝隙连接与卵母细胞进行细胞之间的“对话”,并调控卵母细胞的生长与成熟。卵母细胞成熟后发生排卵,卵丘细胞通过自身及分泌作用影响着精卵结合及雄原核的形成。在哺乳动物的研究中发现,卵丘细胞与胚胎共培养能够克服发育阻滞,并对胚胎的发育起很好的支持作用。随着人类辅助生殖技术的发展,学者们越来越重视在基因及转录水平上对卵丘细胞的检测,从而对卵子及胚胎质量进行预测。综述颗粒细胞在卵子生长与成熟、受精及胚胎发育等方面的作用。     

配子捐赠指征界定与伦理思考

新兴发展的辅助生殖技术（ART）为不孕不育者带来了福音,亦为遗传病患者与高龄女性带来了为人母的希望。由于ART所涉及的不仅有不孕不育夫妇的利益,尚有家庭、后代及社会问题,因此伴随着供精、供卵等新的ART衍生技术的实施,相关伦理问题日益突出,ART的应用与发展面临新的挑战。探讨卵子捐赠以及几种特殊情况下供精指征。