Supplementary annual report of Council, 1982-1983. Appendix VI: Interim report on human in vitro fertilisation and embryo replacement and transfer

The working group set up by the British Medical Association to study the ethical aspects of in vitro fertilization has completed its interim report, issued here as an appendix to the BMA's Annual Report of Council, 1982-1983. Among the issues considered were the acceptability of in vitro fertilization and embryo transfer as a treatment for infertility, the importance of informed consent and detailed recordkeeping, the use of donor ova and sperm, and research with excess embryos. In an addendum, the working group concurred with the BMA's 1979 recommendation that children born as a result of artificial insemination be considered to be legitimate.     

玻璃化冷冻人卵母细胞体外受精－胚胎移植9例临床分析

目的：探讨人卵母细胞玻璃化冷冻在体外受精－胚胎移植（ IVF-ET）中的应用效果。方法行IVF-ET 治疗、因取卵日男方因素无法进行受精的患者9例,对其成熟卵子进行玻璃化冷冻保存,解冻后进行卵胞浆内显微授精（ICSI）,记录卵子复苏、受精、卵裂及移植后妊娠情况。结果9例患者共解冻复苏58枚卵子,复苏率68．2％（58／85）;复苏后进行ICSI的成熟卵子中,受精40枚,形成胚胎34枚,受精率69．0％,卵裂率85．0％。7个移植周期共移植胚胎15枚,临床妊娠3例,临床妊娠率42．9％。结论人卵母细胞玻璃化冷冻可以获得一定的临床妊娠率,在IVF-ET中可作为临床治疗的选择之一。     

睾丸取精卵浆内显微注射受精治疗无精症的疗效分析

目的：评价采用睾丸取精卵浆内显微注射法治疗无精症的疗效。方法：对2000年7月--2002年7月来我所就诊的35例患者，女方取卵当日，男方取精。手术时，患者取平卧位，术野常规消毒，用1％利多卡因局麻，新鲜人体睾丸组织通过切口活检取材，立即放入装有培养液的培养皿中。取部分睾丸组织在体视显微镜下，用无菌针头划开曲精小管，在显微镜下检查是否存在精子。如果找到精子，将存有睾丸组织的培养皿放入培养箱，直到注射前取出。B超引导下经阴道取卵，常规去卵丘结构处理。在倒置显微镜下进行胞浆内单精子注射(ICSI)。注射后15--18小时检查有无原核，48小时观察卵裂，经阴道宫腔内移植质优胚胎1--3个。移植后11天进行妊娠试验检查，7周进行B超检查，了解有无胎心搏动。结果：35例无精症患者全部在睾丸曲精小管中找到精子。女方取卵466个(5--32个)，其中MⅡ期卵367个，显微注射337个，有270个受精卵见到两个原核，受精率为80．11％，35对全部获得受精卵。临床妊娠率为40％。结论：睾丸取精卵浆内显微注射技术是目前治疗无精症最有效的方法。     

诱变剂诱导的精子染色体异常及其对胚胎发育的影响

以环磷酰胺处理雄性小鼠后每周与超排卵雌鼠交配,制备分析受精卵染色体,持续整个生精周期（７周）。处理组小鼠雄原核染色体出现多种畸形,最敏感期在处理后第８天,相当于诱变剂作用于睾丸精子期。处理后第８天的小鼠精子体外受精试验表明,睾丸精子期受环磷酰胺作用后的精子其活力和密度未受显著影响,但其对卵子的受精能力降低,胚胎发育受到显著阻滞（Ｐ＜０．０１）。     

Physiological Roles of Platelet-activating Factor in Mammalian and Human Reproduction

This review described origination, biosynthesis and functions of platelet-activating factor （PAF） in the reproductive system of mammals and human beings. The article mainly focused on biological roles of the phospholipid mediator in sperm fertilization and embryonic implantation. As an autocrine product of sperm and embryos, PAF markedly stimulates sperm motility and fertilization and serves as a capacitation factor in a ligand-receptor manner, After fertilization, embryo-derived PAF improves its own development, especially from fertilized ova to blastocyst stage and is thought to act as an embryo growth factor in the same manner as on sperm. Its mechanism of action was also clarified. At the end, it was presented some advances in its clinical application, followed by discussion of some issues possibly concerning in its current application.     

FVB小鼠精子冷冻及体外受精的研究

目的：探索FVB小鼠及FVB遗传工程小鼠种系保存的新途径.方法：用孕马血清和人绒毛膜促性腺激素（HCG）超排FVB小鼠,并分别用复苏精子和新鲜精子进行体外受精并将受精卵移植到假孕雌鼠输卵管中,比较两种精子的受精率;对注射HCG后13,15,17h不同时间采集FVB小鼠的卵母细胞进行体外受精,比较其受精率及体外培养至2细胞情况.结果：新鲜精子的受精率（62.4%）略高于冷冻复苏精子的受精率（58.1%）;在注射HCG 13～15 h后取FVB小鼠卵母细胞其受精率效果较好,受精率（60.2%、61.6%）显著高于17h取卵的受精率（51%）,但13,15,17 h采集卵母细胞体外受精后胚胎发育至2细胞及异常卵数无显著性差异.结论：本研究将为FVB小鼠及FVB遗传工程小鼠的种系保存、繁殖提供了基础.     

GENERATION AND PARTIAL CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST SOLUBILIZED MEMBRANE FRACTION OF HUMAN SPERMATOZOA

Human sperm membrane antigens extracted by deoxycholate (DOC) were used to immunizeBALB/c mice.Hybrid cell lines secreting sperm-specific monoclonal antibodies were generatedby cell fusion in a semi-solid medium and screened by indirect immunofluorescent assay usinglive and methanol-fixed sperm.Out of 850 hybrid clones from cell fusion,28 were shownto secrete sperm-specific antibodies which reacted with the acrosome,equatorial segment,whole surface plasma membrane or tail of spermatozoa.Finally,seven hybrid cell lineswere established and shown to secrete monoclonal antibodies which had no cross-reactivitywith arty human tissues other than testis and sperm.The majority were also shown toinhibit fertilization of mouse oocytes in vitro and human sperm penetration of zona-freehamster ova.Western blot analysis revealed that some of these antibodies reacted withsperm membrane antigens of distinct molecular size.     

Ca^2＋对人精子穿透无透明带金黄地鼠卵的影响

应用人精子穿透无透明带金黄地鼠卵（SPA）技术，在BWW培养液中分别以不同浓度的Ca^ （1．71mmol,3.42mmol,5.13mmol,1.71mmol和ETA0．50mmol,1.71mmol和EGTA1．71mmol),对10例人精液标本进行分析，研究Ca^ 对人精子体外获能和穿透无透明带金黄地鼠卵的生物学效应，同时用透射电镜观察精子的亚细胞结构变化。结果表明：对照组（Ca^ 1．71mmol)穿透率为82．2％，而试验组的穿透率显著降低（P＜0．01），分别为9．5％，0，17．2％，0。Ca^ 对人精子亚微结构的影响主要发生在精子的顶体和颈部。本试验初步表明Ca^ 与人精子获能和顶体反应存在着一定的依存关系，Ca^ 浓度过高（＞1．71mmol）或过低（＜1．71mmol)，对精子获和穿透能力都有明显的抑制作用。     

大剂量γ射线照射诱发人精子与淋巴细胞染色体畸变研究

目的 :探讨大剂量60 Coγ射线事故照射和离体照射对人精子与淋巴细胞染色体畸变的影响。方法 :应用人精子穿透金黄地鼠卵获得精子中期分裂相和微量全血培养法制备淋巴细胞中期分裂相 ,分析两例在60 Co辐射事故中全身受到大剂量γ射线照射的急性放射病患者照后 6( 7)年的精子和淋巴细胞染色体畸变 ,以及60 Coγ射线离体照射人精子与淋巴细胞诱发染色体畸变 ,并对全身照射与模拟离体照射进行比较。结果 :两例事故受照患者的畸变精子率显著高于对照组 ,亦显著高于同期淋巴细胞染色体畸变水平 ;两患者精子染色体断裂型畸变是重接型的 2倍 ,而淋巴细胞重接型畸变是断裂型的 2倍 ,两患者的Y 精子与X 精子的比例明显增高 ( 1 60∶1)。在离体照射条件下 ,人精子和淋巴细胞的畸变细胞率基本相同 ,均与照射剂量呈正相关 ;结构畸变类型明显不同 ,精子染色体断裂型畸变是重接型的 8倍 ,淋巴细胞重接型畸变是断裂型的 4倍 ;Y 精子与X 精子的比例为 1∶1。在相同剂量点 ,模拟照射组的人精子染色体畸变率显著高于全身照射组 ,两受照组精子染色体均以断裂型畸变为主 ,但事故照射组精子重接型畸变的比例显著高于离体照射组。结论 :在全身照射情况下 ,电离辐射对生殖细胞影响的重点是精原干细胞 ,而且在精子发生的过程中有     

五加皮遗传毒性实验研究

目的：探讨五加皮对小鼠体细胞和生殖细胞的遗传毒性。方法：采用小鼠骨髓细胞微核试验（ Ｍ Ｎ Ｔ）、骨髓和睾丸染色体畸变（ Ｃ Ａ）试验以及精子畸形试验。结果：五加皮诱发的微核、染色体畸变和精子畸形均低于正常对照,特别是精子畸形率明显低于正常对照（ Ｐ＜ ００１）。结论：五加皮对小鼠体细胞和生殖细胞均无诱变损伤作用。     

序贯培养体系和单一培养体系对人卵母细胞受精和早期胚胎发育的影响

目的：比较在体外受精/卵胞质内单精子显微注射（IVF/ICSI）治疗周期中序贯培养体系和单一培养体系对人类早期胚胎发育的影响，为人类辅助生殖技术中胚胎培养体系的选择和评估提供参考。方法：选取155例行IVF/ICSI助孕的患者，将同一个患者的卵子分为2组，分别在Vitrolife的序贯培养体系G-IVF/G1/G2培养液（序贯培养组）和Irvine的单一培养体系CSCC培养液（单一培养组）中进行体外受精和胚胎培养，观察2组不同培养体系中胚胎受精和胚胎早期发育情况。结果：与单一培养组比较，序贯培养组受精率、双原核（2PN）受精率和多核受精率差异均无统计学意义（P>0.05）。在IVF患者中，序贯培养组和单一培养组的卵裂率、可用胚胎率、优质胚胎率和囊胚形成率比较差异均无统计学意义（P>0.05）；在ICSI患者中，序贯培养组和单一培养组的卵裂率、可用胚胎率和囊胚形成率比较差异均无统计学意义（P>0.05），但序贯培养组优质胚胎率明显高于单一培养组（P=0.015）。在IVF和ICSI患者中，单一培养组第3天胚胎致密化的比例均明显高于序贯培养组（P=0.001）。在序贯培养组中胚胎形态表面光滑均匀，单一培养组中胚胎表面较为粗糙且有颗粒状。结论：G-IVF/G1/G2序贯培养体系和CSCC单一培养体系对人卵母细胞受精和早期胚胎发育的影响无明显差异。     

Should we clone human beings? Cloning as a source of tissue for transplantation.

The most publicly justifiable application of human cloning, if there is one at all, is to provide self-compatible cells or tissues for medical use, especially transplantation. Some have argued that this raises no new ethical issues above those raised by any form of embryo experimentation. I argue that this research is less morally problematic than other embryo research. Indeed, it is not merely morally permissible but morally required that we employ cloning to produce embryos or fetuses for the sake of providing cells, tissues or even organs for therapy, followed by abortion of the embryo or fetus.     

转基因技术和冷冻胚胎技术的实验数据建立

目的 建立一系列转基因技术和冷冻胚胎技术的实验室数据库。方法 a)显微注射法注射1864个1-细胞期受精卵,注射后存活的1283个胚胎进行胚胎移植,获得20只阳性转基因动物。b)设计不同单位的激素剂量(PMSG HCG)获得每个品系的小鼠平均排卵量和最佳激素剂量。e)采用常规法和玻璃化法进行胚胎冷冻保存,并就囊胚和桑椹胚两个不同时期进行复苏成活率的比较。结果 和讨论a)建立转基因技术和胚胎冷冻保存的实验数据库(随着实验技术的不断提高还将不断的完善)。胚胎显微注射存活率为68．8％;转基因阳性鼠出生率为16．16％;基因整合率为1．56％。b)激素诱发5种品系小鼠的平均超排卵数为：C57BL/6J(日本)15．6;C57BL/6J 19．6;BDF1(日本)21．4;C3H(日本)14．8;KM 30。e)胚胎冷冻保存复苏,出生幼子存活率为30％～31％。     

Cytogenetic effect of thioridazine hydrochloride in mice and in human lymphocyte chromosomes

Thioridazine hydrochloride, a drug of the phenothiazine group with a piperidine side chain is widely used in human medicine as a psycho-relaxant and anxiolytic agent. The mutagenic effects of thioridazine hydrochloride were studied in Swiss albino mice. The animals were treated with 4, 6 and 8 mg/kg of thioridazine hydrochloride for the micronucleus test, analysis of chromosomes in germ cells, and for sperm morphology assay. Human lymphocyte cultures were treated in vitro with 0.75, 1.25 and 1.75 mg/culture of thioridazine hydrochloride for 24, 48 and 72 hours. The study showed no significant increase in the frequency of micronuclei, chromosomal aberrations in germ cells and sperm head abnormalities in mice, or chromosomal aberrations in human lymphocyte cultures.     

Mummy was a fetus: motherhood and fetal ovarian transplantation.

Infertility affects 15 per cent of the world's couples. Research at Edinburgh University has been directed at transplanting fetal ovarian tissue into infertile women, thus enabling them to bear children. Fetal ovary transplantation (FOT) has generated substantial controversy; in fact, one ethicist deemed the procedure 'so grotesque as to be unbelievable' (1). Some have suggested that fetal eggs may harbour unknown chromosomal abnormalities: however, there is no evidence that these eggs possess a higher incidence of genetic anomaly than ova found in a healthy adult female. There is also concern that fetal egg children will be psychologically harmed by the knowledge of their special conceptual status. It will be demonstrated that special conceptual status in and of itself does not determine developmental success. Rather, psychological well-being is dependent upon how the family and child cope with the unique challenges inherent in FOT. Lastly, though considering FOT a legitimate method of family building, given the global population crisis the wisdom of procreational rights will be challenged. Inherent to this challenge is a re-evaluation of the treatment of infertility as a significant disease necessitating remedy.     

Effects of electromagnetic radiation from a cellular phone on human sperm motility: an in vitro study

BACKGROUND: There has been growing public concern on the effects of electromagnetic radiation (EMR) emitted by cellular phones on human health. Many studies have recently been published on this topic. However, possible consequences of the cellular phone usage on human sperm parameters have not been investigated adequately. METHODS: A total number of 27 males were enrolled in the study. The semen sample obtained from each participant was divided equally into two parts. One of the specimens was exposed to EMR emitted by an activated 900 MHz cellular phone, whereas the other was not. The concentration and motility of the specimens were compared to analyze the effects of EMR. Assessment of sperm movement in all specimens was performed using four criteria: (A) rapid progressive, (B) slow progressive, (C) nonprogressive, (D) no motility. RESULTS: Statistically significant changes were observed in the rapid progressive, slow progressive and no-motility categories of sperm movement. EMR exposure caused a subtle decrease in the rapid progressive and slow progressive sperm movement. It also caused an increase in the no-motility category of sperm movement. There was no statistically significant difference in the sperm concentration between two groups. CONCLUSIONS: These data suggest that EMR emitted by cellular phone influences human sperm motility. In addition to these acute adverse effects of EMR on sperm motility, long-term EMR exposure may lead to behavioral or structural changes of the male germ cell. These effects may be observed later in life, and they are to be investigated more seriously.     

精子DNA碎片化指数与体外受精胚胎移植结局的关系

目的: 探究精子DNA碎片化指数(DNA fragmentation index,DFI)与体外受精胚胎移植的实验室指标和临床结局的关系。方法: 选取在本院生殖中心就诊且符合纳入标准的90对夫妇为研究对象,以DFI值30％分为正常组、低损伤组、高损伤组。统计男女双方年龄、男方精液常规参数、女方获卵数、受精率、卵裂率、优胎率、囊胚形成率、妊娠率和流产率,分析3组之间各指标的差异。结果： 3组的男女双方年龄、男方精液常规指标、女方获卵数无明显差异(P>0.05)。3组间受精率、卵裂率差异亦无统计学意义(P>0.05);高损伤组的优胎率(59.65％±28.93％)和囊胚形成率(13.49％±17.74％)较正常组(74.34％±21.56％,31.27％±20.54％)明显降低(P0.05)。 结论： DFI值的升高可能与优胎率、囊胚形成率以及妊娠率的降低有关,而与受精率、卵裂率和流产率无明显关联。     

Gestational surrogacy for a human immunodeficiency virus seropositive sperm donor: what are the ethics?

Clinics that provide assisted reproductive technology (ART) are guided by general guidelines set forth by the American Society for Reproductive Medicine and its Ethics Committee and are free to set their own policies within those guidelines. This article presents a case in which a university clinic was presented with a novel request. A same-sex male couple, both positive for the human immunodeficiency virus (HIV), asked to use one of the couple's sperm to establish a pregnancy in an unrelated gestational surrogate through in vitro fertilization, intracytoplasmic sperm injection, and embryo transfer. The couple's argument in favor of such a plan was that no documented case of HIV seroconversion had so far occurred in recipients of gametes from HIV-positive donors. Since gestational surrogates routinely accept the risks inherent in pregnancy and childbearing, an informed surrogate should be allowed to accept the risks of such an arrangement. They further argued that if no clinic were willing to provide such services, data regarding seroconversion would never be obtained. The university ethics committee examined the fertility clinic's policies and found the clinic's refusal to provide such services to be completely consistent with its policy that allows providing services to HIV-discordant couples, same-sex couples, and gestational surrogates, but that always acts to protect the surrogate from exposure to infectious risk.     

Evaluation of Endotoxin in Culture Medium for Human in vitro Fertilization

To determine whether the presence of bacterial endotoxin in the commercial culture media utilized for human in vitro fertilization （IVF）, and evaluate the difference in detecting endotoxin in culture medium between the human sperm motility assay and the 2-cell mouse embryo assay. Methods Thirty-six batches of culture media commonly used in IVF laboratories from 3 manufacturers were determined for thepresence ofendotoxin before using the medium for the assisted reproductive programs （group A）. After being used, 25 specimens among above media were also tested （group B）. The chromogenic limulus amoebocyte lysate （LAL） test was used for quantification the content of endotoxin. In addition, the human sperm motility assay was compared with the 2-cell mouse embryo assay to evaluate the difference in detecting endotoxin in culture medium. Results Endotoxin was not detected in group A. However, 2 samples were positive in group B, Sperm did not show significant change in motility in group A during 24 h of incubation when compared with the control （P〉0. 05）. However, in group A the 2-cell embryo development to blastocyst was suppressed in 3 batches of media. Conclusions Regular screening of each batch of culture medium should be performed if possible although there was no evidence of endotoxin contamination in commercially prepared pre-tested media. Culture environment should be stringently controlled in case the medium is polluted. The sensitivity of the sperm motility assay was lower than that of the mouse embryo assay for detecting low levels of endotoxin or toxic compounds in the medium.     

Recovery of human ova from the uterine tubes

To study the time of ovulation in the menstrual cycle, an investigation was planned to recover ova from the uterine tubes, to correlate their condition with the menstrual history and stage of development of the early corpora lutea from which these ova had been extruded, and to quantitatively analyze the ovarian hormone level in tissues of the human ovary. The preliminary report records the recovery of 7 ova from the uterine tubes. 1 case involved twin ova where each ovary contained an early corpus luteum. Another involved internal migration of the ovum from the left ovary to the right tube. One ova was recovered on the 12th, 4 on the 15th, and 2 on the 16th day of the menstrual cycle. The last 2 showed signs of degeneration. The corpora lutea of each ova were examined and described. The ova, uterine tubes, corpora lutea, and some endometrium are being prepared for histologic st udy. A series of analyses of hormone content of human ovarian tissues is also being accumulated.