Pregnancy derived from human zygote pronuclear transfer in a patient who had arrested embryos after IVF.

Nuclear transfer of an oocyte into the cytoplasm of another enucleated oocyte has shown that embryogenesis and implantation are influenced by cytoplasmic factors. We report a case of a 30-year-old nulligravida woman who had two failed IVF cycles characterized by all her embryos arresting at the two-cell stage and ultimately had pronuclear transfer using donor oocytes. After her third IVF cycle, eight out of 12 patient oocytes and 12 out of 15 donor oocytes were fertilized.

Nonprofessional phagocytosis in trophectoderm cells of human preimplantation blastocysts.

Unlabelled: Trophoblast phagocytosis has been considered important in pregnancy. However, whether human preimplantation blastocysts possess phagocytic activity is still unclear. In this study, we determined the phagocytosis potential in human trophectoderm cells of blastocysts prior to implantation.

[Effect of domestic highly purified urinary follicle stimulating hormone on outcomes of in vitro fertilization-embryo transfer in controlled ovarian stimulation].

Objective: To investigate the effect of domestic urine-derived high-purity follicle- stimulating hormone (HP-FSH, Lishenbao) on the outcome of in vitro fertilization(IVF) embryo transfer (ET) in controlled ovarian stimulation (COS).

Methods: From 1 September 2010 to 31 March 2011, total of 3178 infertility patients from 14 Reproductive Center with IVF or intracytoplasmic sperm injection (ICSI) indications who accepted first IVF or ICSI cycle were studied retrospectively. Their causes of infertility include all infertility factors except ovulatory dysfunction infertility and uterine factor infertility.

A highly sensitive electrochemiluminescence immunoassay for detecting human embryonic human chorionic gonadotropin in spent embryo culture media during IVF-ET cycle.

Purpose: To investigate the stability and repeatability of electrochemiluminescence immunoassay (ECLIA) for beta-hCG detection in embryo spent culture media. To evaluate the correlation between the viability of preimplantation embryo and beta-hCG profile by the new assay.

Methods: In a retrospective study, a total of 357 spent culture media from day1 to day5 were individually collected and quantified by ECLIA.

Preimplantation genetic diagnosis for α-and β-double thalassemia.

Purpose: To evaluate the use of multiple displacement amplification (MDA) for preimplantation genetic diagnosis (PGD) of α- and β-double thalassemia.

Method: Whole genome of a single cell was directly amplified using MDA and its products were used as templates in fluorescent gap polymerase chain reaction (PCR) analysis of α-thalassemia and in PCR-reverse dot blot analysis, singleplex fluorescent PCR of β-28 and CD17 mutation and HumTH01 for β-thalassemia.

Results: 1) MDA from single cell could produce enough DNA templates for the detection of both α and β-thalassemia; 2) The established MDA-PGD protocol for α- and β-double thalassemia was successfully applied in PGD of six embryos, among which, three were transferred, but no pregnancy ensued.

[Clinical analysis of 100 preimplantation genetic diagnosis cycles].

Objective: To investigate influence of chromosomal translocations on early embryo development and to evaluate the efficacy and feasibility of preimplantation genetic diagnosis (PGD) techniques through clinical analysis on PGD cycles.

Methods: Embryo development, efficacy of PGD and clinical outcome of 100 cycles were studied retrospectively, including 23 cycles with Robertsonian translocations, 19 cycles with reciprocal translocations, and 58 cycles for α-Thalassaemia.

Results: Among 354 embryos biopsied by PGD for translocations, 321 (90.

Supraphysiological estrogen levels adversely impact proliferation and histone modification in human embryonic stem cells: possible implications for controlled ovarian hyperstimulation assisted pregnancy.

Objective: Controlled ovarian hyperstimulation (COH) results in supraphysiologic levels of maternal serum estradiol (E(2)) during the luteal phase, thus promoting oocyte production at unknown risk to the subsequently developing embryo. Human embryonic stem cells (hESCs) have been identified as a model system to assess the impact of COH on early embryonic development, specifically 17β-estradiol mediated effects on proliferation, gene expression, and histone modification.

Study Design: Cell proliferation and associated factors, such as HDAC1, as well as histone modification patterns were evaluated in ERα and β expressing hESCs after exposure to 17β-estradiol (1×10(-10) M to 1×10(-7) M), as well as in an untreated control.

Improved nude mouse models for green fluorescence human endometriosis.

Aim: To establish an improved noninvasive fluorescent animal model for endometriosis.

Material And Methods: Adenovirus encoding enhanced green fluorescent protein (Ad-eGFP) was used to transfect primary culture endometrial glandular cells and stromal cells (purified cell transfection and mixed injection, Group 1) as well as endometrial fragments (tissues transfection and injection, Group 2). Transfection results were compared between the cells and tissues in vitro.

Imprinting and Promoter Usage of Insulin-Like Growth Factor II in Twin Discordant Placenta.

Case reports from infant twins suggest that abnormal genomic imprinting may be one of the important causes of twin discordance, but it is unknown whether abnormal genomic imprinting occurs in the placenta. Therefore, we sought to determine the relationship between the imprinting of insulin-like growth factor II (IGF-II) in placenta and twin discordance. We analyzed the imprinting and promoter usage of IGF-II in placenta of normal twins (T0 group), weight discordance (T1 group), and phenotype discordance (T2 group).

High follicle-stimulating hormone increases aneuploidy in human oocytes matured in vitro.

Objective: To study the effect of FSH on the aneuploidy risk of human oocytes matured in vitro.

Design: Prospective study.

Setting: Hospital-based IVF center.

[Evaluation of the fidelity of multiple displacement amplification from small number of cells].

Objective: To evaluate the fidelity of multiple displacement amplification (MDA) from small number of cells (1-10 cells) by 10K 2.0 SNP mapping array.

Methods: A fibroblast cell line (Tri-18; GM02732, 47, XY, +18) was used as the template, and 6 groups were set up in the study.

Preimplantation genetic diagnosis for alpha-thalassaemia in China.

Purpose: To report the usage of PGD for alpha-thalassaemia with the - -(SEA) genotype.

Method: A PGD protocol using fluorescent gap PCR was performed for 51 cycles on 43 couples with the - -(SEA) genotype. Allele drop-out and amplification failure rates were retrospectively analyzed.

Mechanically expanding the zona pellucida of human frozen thawed embryos: a new method of assisted hatching.

Objective: To determine whether a new assisted hatching (AH) method increases the implantation and clinical pregnancy rates of frozen-thawed day-3 (D3) embryos.

Design: Prospective study.

Setting: A university hospital in vitro fertilization (IVF) program.

Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array.

The scarce amount of DNA contained in a single cell is a limiting factor for clinical application of preimplantation genetic diagnosis mainly due to the risk of misdiagnosis caused by allele dropout and the difficulty in obtaining copy number variations in all 23 pairs of chromosomes. Multiple displacement amplification (MDA) has been reported to generate large quantity of products from small amount of templates. Here, we evaluated the fidelity of whole-genome amplification MDA from single or a few cells and determined the accuracy of chromosome copy number assessment on these MDA products using an Affymetrix 10K 2.

RNA interference targeting of sphingomyelin phosphodiesterase 1 protects human granulosa cells from apoptosis.

Aim: Sphingomyelin phosphodiesterase 1 (SMPD1) plays an essential role in initiating the female germ cell death signal. To evaluate whether RNA interference has potential as a new approach in germ cell protection, we tested the effect of SMPD1 knockdown on human granulosa cells in vitro.

Methods: We designed and synthesized three small interference RNA (siRNA) sequences targeted on SMPD1 and transfected them into human luteinizing granulosa cells (hGC) in vitro.

Expression of certain HLA-I types in cleavage-stage embryos.

This study examined the expression of human leukocyte antigen (HLA)-G and HLA-I (which includes HLA-A, -B, -C, -E and -F, but is without HLA-G) in the cleavage embryo and its supernatant, and related the results to embryo development including growth rate and grade. In total, 136 day-3 cleavage embryos were used for detection of HLA-G and 24 embryos for HLA-I without HLA-G by immunohistochemistry. The expression of HLA-I was examined by western blot in the lysates of a further 63 day-3 cleavage embryos; soluble HLA-I in the culture supernatant of embryos with detectable HLA-I was also examined by western blot.

[Efficiency comparison between two preimplantation genetic diagnostic methods for chromosomal translocation carriers].

Objective: To compare the diagnostic efficiency between blastomere preimplantation genetic diagnosis (PGD) and polar body PGD for chromosomal translocation carriers.

Methods: Group A had 8 cycles using whole painting probes for the first polar body diagnosis, while group B had 29 cycles using two subtelomeric probes and one centromeric probe for the blastomere diagnosis.

Results: The fertilization rate of group A was significantly lower than group B [66.

[Variance of serum prolactin in controlled ovarian stimulation].

Objective: To investigate the variance of peripheral blood prolactin (PRL) in controlled ovarian stimulation.

Methods: Seventy-two patients, with totally 106 cycles receiving a long protocol of gonadotropin-releasing hormone agonist combined with gonadotropin (Gn) were randomly enrolled in this retrospective study. During controlled ovarian stimulation, peripheral blood hormones were measured by chemiluminescent microparticle immunoassay.

Preimplantation genetic diagnosis for Duchenne muscular dystrophy by multiple displacement amplification.

Objective: To evaluate the use of multiple displacement amplification (MDA) in preimplantation genetic diagnosis (PGD) for female carriers with Duchenne muscular dystrophy (DMD).

Design: MDA was used to amplify a whole genome of single cells. Following the setup on single cells, the test was applied in two clinical cases of PGD.

In vitro-matured rat oocytes have low mitochondrial deoxyribonucleic acid and adenosine triphosphate contents and have abnormal mitochondrial redistribution.

Objective: To study the development and function of mitochondria in in vitro-matured rat oocytes derived from follicles of different sizes.

Design: Experimental animal study.

Setting: Department of Anatomy at the University of Hong Kong.

Effect of small interfering RNAs of cytochrome P450 17alpha-hydroxylase/17,20-lyase (CYP17) on androgen biosynthesis in theca cells.

Polycystic ovary syndrome (PCOS) is associated with a variety of endocrinologic and metabolic abnormalities, with clinical features of hyperandrogenism and hyperandrogenemia. Cytochrome P450 17alpha-hydroxylase/17,20-lyase (CYP17) is critical in androgen biosynthesis, and CYP17 mRNA expression was proven augmented in PCOS theca cells. To demonstrate whether RNA interference (RNAi) could lower the androgen concentration in theca cells, small interfering RNA (siRNA) targeting the CYP17 gene was co-cultured with exogenous CYP17 in HeLa cells and endogenous CYP17 of theca cells.

Effects of cilostamide and forskolin on the meiotic resumption and embryonic development of immature human oocytes.

Background: In an attempt to allow for acquisition of oocyte cytoplasmic maturation, PDE3 specific inhibitor, cilostamide and adenylate cyclase activator, forskolin were used to extend pre-maturation culture of immature human oocytes.

Methods: Cumulus-oocyte complexes retrieved from unstimulated ovaries were continuously cultured under 20 microM cilostamide or 50 microM forskolin, alone or in combination for 6, 12, 24 or 48 h, respectively. Levels of intercellular gap junction communication (GJC) and maturational status were examined at these designated time points.

Bulk vitrification of human embryonic stem cells.

Background: The traditional vitrification method cannot keep up with the increased culture and propagation efficiency required to cryopreserve large quantities of vigorously proliferating human embryonic stem (HES) cells. In this study, we describe a newly invented vitrification carrier for cryopreserving large amount of HES cells and evaluate whether this bulk vitrification (BV) method is as effective as the popular open-pulled straw (OPS) vitrification method.

Methods: HES cell clumps were harvested after passage and transferred to a cell strainer; only those clumps with a diameter more than 70 microm were included in the study and randomly selected to be cryopreserved by the BV method, OPS vitrification or slow freezing method.