Effects of Growth Hormone Supplementation in Patients Undergoing IVF/ICSI-ET with Poor Ovarian Response to Gonadotropin

Objective To analyze the effects of growth hormone （GH） supplementation during IVF/ ICSI-ET in Chinese patients who had prior IVF cycle with poor response to gonadot- ropin （Gn）. Methods Ovulation was stimulated in 389 consecutive patients who all had poor ovarian response, among them, 102 patients （GH cycle） received 4 IU GH and the other 287 patients （non-GH cycle） underwent IVF without GH. Fisher＇s exact test, Chi square test and Student＇s t-test were used to analyze IVF/ICSI-ET outcomes. Results After GH treatment, 102 patients had significantly more large- and medium- sized follicles, oocytes retrieved, 2 pronucleus oocytes, metaphase H stage （M~I） oocytes, and high-quality embryos than in previous cycles without GH. However, the number of embryos transferred, clinical pregnancy rate, transfer rate and biochemical pregnancy rate were not significantly different. Furthermore, the 102 patients given GH had significantly lower luteinizing hormone levels and biochemical pregnancy rates; thicker endometrium and more Gn administration days; and more large- and medium-sized follicles and M~I oocytes than 287 other patients undergoing IVF/ICSI-ET without GH. However, these groups did not differ significantly in clinical pregnancies, high-quality embryos, Mn oocytes, and embryo implantation rates. Conclusion GH may improve some IVF/ICSI-ET outcomes for women with poor ovarian response.     

波尔山羊腹腔内窥镜鲜胚移植试验

2002年10-11月在山东省东营市蓝海集团绵野公司种羊场使用两种超排药物对40只经产波尔山羊进行超排处理．供体羊有两只未发情．超排有效率为95％(38／40)。共回收胚胎388枚．其中可用胚311枚．每只获胚胎10．21枚(388／38)，平均可用胚8．18枚(311／38)，可用胚率80．15％(311／388)。同期处理受体399只，有320只发情，同期率为80．20％．运用腹腔内窥镜技术移植受体267只．其中44只供体移植双胚。经试情150只受体未返情，受胎率为56．18％(150／267)。     

诱发超排卵时卵巢过度刺激综合征的预测和防治

诱发超排卵条件下进行16例配子输卵管内移植（GIFT），共发生5例卵巢过度刺激综合征（OHSS），OHSS及重度OHSS发生率分别为31．3％（5／16）及6．3％（1／16）。结果提示：OHSS的发生主要与hMG、hCG用量尤其是黄体期hCG用量有密切关系，注射hCG前血巳水平≥1500ng／L及卵泡数≥10个为发生OHSS的高危因素；妊娠可以促进或加重OHSS；通过连续阴道B超监测卵泡生长；调节hMG用量；结合注射hCG前血E2水平决定黄体期hCG用量，可有效地防止OHSS发生。     

IL-6对超排卵后小鼠受精卵发育的影响

目的 探讨IL-6对超排卵后小鼠受精卵发育到两细胞的影响.方法 本实验分为3个部分.①IL-6添加实验:80只ICR雌鼠采用随机数字表法分为7组,包括超排卵组(n=60),正常自然周期组(即对照组,n=20).其中超排卵组再按照超排卵后体外培养液中添加IL-6浓度的不同分为超排卵对照组(0pg/ml IL-6组)、1pg/ml IL-6组、5pg/mlIL-6组、10pg/ml IL-6组、25pg/mlIL-6组和50pg/mlIL-6组,每组10只.②IL-6受体抗体(RA)添加实验:90只ICR雌鼠采用随机数字表法分为7组,包括超排卵5组(n=50),正常自然周期2组[包括对照组(n=20)和IL-6受体抗体(100pg/ml)组(n=20)].其中超排卵组按照超排卵后体外培养液中添加不同浓度IL-6及IL-6受体抗体分为超排组、1pg/ml IL-6+RA组、5pg/ml IL-6+RA组、10pg/ml IL-6+RA组、25pg/mlIL-6+RA组,每组10只.对照组小鼠正常自然受孕,超排组小鼠超排卵后受孕,收集受精卵体外培养1d至两细胞,显微镜下观察两细胞形成率.以上两部分实验重复3次.③IL-6免疫荧光表达实验:10只ICR雌鼠采用随机数字表法分为对照组和超排组,每组5只,应用免疫荧光共聚焦法检测对照组和超排组受精卵中IL-6的表达.结果 IL-6添加实验结果显示,超排对照组,1pg/mlIL-6组、25pg/ml IL-6组及50pg/mlIL-6组两细胞形成率明显低于对照组,差异有统计学意义(P=0.023、P=0.026、P=0.000、P=0.000).IL-6受体抗体(RA)添加实验结果显示,与正常对照组比较,超排组两细胞形成率下降(P=0.017);添加IL-6受体抗体的各组两细胞发育率也下降,差异有统计学意义(P=0.000).IL-6免疫荧光表达实验结果显示,超排组受精卵中IL-6表达明显低于对照组.结论 控制性超排卵能够降低受精卵中IL-6的表达,可能是超排卵影响胚胎发育的机制之一.     

GH/tPA双转基因小鼠的制备及其表达分析

目的 旨在获得GH/tPA双转基因小鼠,并比较分析小鼠乳腺中人组织纤溶酶原激活剂（tPA）的表达水平和个体生长发育状况。方法 利用2只tPA单转基因雄性小鼠（P03、P05）与经超数排卵处理的正常非转基因雌性小鼠交配,受精卵显微注射线性化的GH质粒、胚胎移植而获得后代小鼠,PCR检测基因整合情况,ELISA和Western blotting检测比较单、双基因表达tPA水平,监测转基因小鼠不同生长阶段的体质量来分析GH基因对小鼠生长发育的影响。结果 P03系小鼠共获得286枚受精卵,移植受体母鼠后,顺利产仔77只,经PCR检测获得16只tPA单转基因小鼠（7♂,9♀）,13只GH/tPA双转基因小鼠（8♂,5♀）,双基因整合率为16.9%;P05系小鼠共获得175枚受精卵,移植受体母鼠后,顺利产仔34只,经PCR检测获得12只tPA单转基因小鼠（5♂, 7♀）,7只GH/tPA双转基因小鼠（3♂,4♀）,双基因整合率为20.6%。单、双转基因小鼠乳腺中表达tPA的最高量分别为82.5 μg/mL、674 μg/mL,GH/tPA双转基因小鼠乳腺中表达tPA的量明显高于tPA单转基因（P<0.05）。在小鼠的整个生长周期内,单、双转基因与正常非转基因小鼠的体质量增长均没有显著的差异（P>0.05）。结论 本实验成功地制备了GH/tPA双转基因小鼠,结果表明GH基因导入小鼠体内能够显著地提高目的基因tPA的表达水平,且不会对转基因小鼠的生长发育造成任何明显的影响,这为将来制备高表达转基因动物生产医药蛋白及其育种奠定了基础。     

不同超排方法对家兔超数排卵效果的影响

目的 探讨家兔稳定高效的超数排卵方法, 旨在获得充足的优质兔卵母细胞,为随后的相关试验提供可靠的物质保障.方法 尿促性素(HMG)+人绒毛膜促性腺激素(HCG)组:雌兔颈部皮下注射HMG 35 IU/只,每隔24 h注射1次,共2次;孕马血清促性腺激素(PMSG)超排组:雌兔颈部皮下一次性注射PMSG促卵泡生长发育,每...     

In vitro fertilization and embryo transfer in domestic animals: Applications in animals and implications for humans

Extensive research has been conducted in domestic animals, particularly in cattle, in the reproductive technologies of sperm handling, capacitation, and acrosome reaction, superovulation, and embryo handling, sexing, bisection, cryopreservation, and transfer. Because of the economic importance of cattle these technologies have been tested and improved under clinical conditions: The results of employing these procedures are available on tens of thousands of pregnancies and offspring. This information has implications in applying some of the same technologies in human reproduction. The large number of normal progeny produced in cattle after a long prenatal development period, similar to humans, provides some assurance that these technologies, carefully applied, are safe. The basis for these conclusions is documented in the publications cited in this review.     

The Effect of GnRHa Induced Superovulation on Endometrial Morphology and Estrogen Receptor and Progesterone Receptor in Mouse

How to increase the pregnancy rates of IVF-ET projecthas been an urgentcon-cern.Endometrial maturation,embryo quality and their synchronization are the keypoints of a successful blastocyst implantation. Superovulation by gonadotropin-releasing hormoneagon…     

Development and Application of a Sensitive,Second Antibody Format Enzymeimmunoassay (EIA) for Estimation of Plasma FSH in Mithun (Bos frontalis)

Mithun (Bos frontalis) is a semi-wild rare ruminant species. A simple sensitive enzymeimmunoassay suitable for assaying FSH in the blood plasma of mithun is not available which thereby limits our ability to understand this species reproductive processes. Therefore, the aim of this article was to develop a simple and sensitive enzymeimmunoassay (EIA) for estimation of FSH in mithun plasma and apply the assay to understand the estrous cycle and superovulatory process in this species. To accomplish this goal, biotinylated FSH was bridged between streptavidin-peroxidase and immobilized antiserum in a competitive assay. Forty microlitre mithun plasma was used directly in the EIA. The FSH standards were prepared in hormone free plasma and ranged from 5–1280 pg/well/40 μL. The sensitivity of EIA was 5 pg/well FSH, which corresponds to 0.125 ng/mL plasma and the 50% relative binding sensitivity was 90 pg/well/40 μL. Although the shape of the standard curve was not influenced by different plasma volumes viz. 40 and 80 μL, a slight drop in the OD₄₅₀ was observed with the increasing volume of plasma. Parallelism tests conducted between the endogenous mithun FSH and bovine FSH standards showed good homology between them. Plasma FSH estimated using the developed EIA and commercially available FSH EIA kit in the same samples were correlated (r = 0.98) and showed linearity. Both the Intra- and inter-assay CV were below 6%. Recovery of known concentrations of added FSH showed linearity (r = 0.99). The developed EIA was further validated biologically by estimating FSH in cyclic cows for the entire estrous cycle, in mithun heifers administered with GnRH analogues and in mithun cows during superovulatory treatment with FSH. In conclusion, the EIA developed for FSH determination in mithun blood plasma is simple and highly sensitive for estimation of mithun FSH in all physiological conditions.     

The incidence of chromosomal aneuploidy in stimulated and unstimulated (natural) uninseminated human oocytes.

The incidence of chromosomal aneuploidy in human oocytes is higher than for various animal species. Since this estimate for aneuploidies is based on data obtained from in-vitro fertilization (IVF) patients, it is possible that superovulation could be contributing to this phenomenon. In this study we determine the incidence of chromosomal aneuploidy in nonstimulated uninseminated human oocytes donated by IVF patients. Furthermore, we compare this incidence of aneuploidy to that obtained after superovulation using two different protocols for induction of multiple follicular growth. The rate of aneuploidy in non-stimulated oocytes was 20% (4/20). This is not significantly different from the rate of aneuploidy in oocytes obtained after superovulation with clomiphene/human menopausal gonadotrophin (HMG)/(HCG) (15/43 = 35%, chi 2 = 1.11; P > 0.20), buserelin-flare (8/25 = 32%; chi 2 = 0.32; P > 0.05), and the rate of aneuploidy in the total number of superovulated oocytes (23/68 = 34%; chi 2 = 82; P < 0.30). Furthermore, the incidence of chromosome aneuploidy in non-stimulated uninseminated oocytes (20%) was well within the range and not significantly different from that reported in the literature for both superovulated uninseminated oocytes (range, 21-57%; total aneuploidy rate, 67/216 = 31%; P < 0.30) and superovulated inseminated oocytes (range, 3-56%; total aneuploidy, 339/1480 = 23%; P < 0.95). Consequently, the data provide evidence that superovulation protocols used in IVF may not be responsible for the higher rate of aneuploidy in human oocytes. These results are discussed in relation to hypotheses on the occurrence of meiotic non-disjunction.