Search

Background: Alport syndrome (AS) is a hereditary renal basement membrane disease that can lead to end-stage renal disease in young adults. It can be diagnosed by genetic analysis, being mostly caused by mutations in COL4A3, COL­4A4, and COL4A5. To date, there is no radical cure for this disease. Objectives: The aim of this study was to avoid the transmission of AS within an affected family by selecting healthy embryos for uterine transfer. The embryos were identified by preimplantation genetic testing for monogenic disorders (PGT-M). Methods: We used next-generation sequencing (NGS) to identify mutations in the proband and his parents. The results of NGS were confirmed by Sanger sequencing. Targeted NGS combined with targeted single-nucleotide polymorphism haplotyping was used for the in vitro identification of COL4A5 mutations in human embryos to prevent their intergenerational transmission. Results: The c.349_359delGGACCTCAAGG and c.360_361insTGC mutations in COL4A5 were identified in a family affected by X-linked AS. Whole-genome sequencing by NGS with targeted haplotyping was performed on biopsied trophectoderm cells. A healthy baby was born after transfer of a single freeze-thawed blastocyst. Conclusions: The use of targeted NGS for identifying diagnostic markers combined with targeted haplotyping is an easy and efficient PGT-M method for preventing intergenerational transmission of AS.

Galectin-1 is highly expressed in blastocysts and trophoblast giant cells during implantation, and dysregulated galectin-1 is associated with many pregnancy-related abnormalities. Elevated galectin-1 contributes to cancer cells invasion. Here, we found that galectin-1 is expressed in mouse oocytes, preimplantation embryos (all stages), and trophoblast stem (TS) cells. Peak levels of galectin-1 mRNA and protein were detected on day 4 and day 5 after the induction of TS cells differentiation. Overexpression of galectin-1 increased TS cells migration and invasion, whereas knockdown of galectin-1 attenuated these effects. Additionally, knockdown of galectin-1 in TS cells decreased the expression of matrix metalloproteinase (MMP) 2/9, ZEB-1, Snail, N-cadherin, TGF-β, Nodal, and phospho-Smad2/3, whereas the expression of E-cadherin was increased. In contrast, overexpression of galectin-1 in TS cells increased the expression of MMP2/9, ZEB-1, and N-cadherin, whereas the expression of E-cadherin was decreased. These findings suggest a potential role of galectin-1 in the differentiation of mouse TS cells.

Introduction A systematic review and meta-analysis was carried out to assess the clinical effectiveness of acupuncture in treating polycystic ovarian syndrome (PCOS). Methods RCTs that compared either acupuncture with no/sham (placebo) acupuncture or a certain therapy with acupuncture added in the treatment of PCOS were included in the review. Measures of treatment effectiveness were the pooled odds ratios (OR) for women with PCOS having acupuncture compared with women in the control group for the recovery of menstrual cycles, standardized mean difference (SMD) for body mass index (BMI), fasting insulin (FINS), fasting plasma glucose (FPG), luteinizing hormone (LH), follicle stimulating hormone (FSH), and the ratio of LH/FSH. Results A total of nine RCTs (531 women) met criteria for inclusion into the systematic review. Using the random effects model, pooling of the effect estimates from all RCTs showed recovery of menstrual cycles (OR = 0.20, 95% CI: 0.09–0.41, P P = 0.04), and LH (SMD = −0.39, 95% CI: −0.65 to −0.12, P P > 0.05). Conclusions Acupuncture appears to significantly improve the recovery of the menstrual cycles and decrease the levels of BMI and LH in women with PCOS. However, the findings should be interpreted with caution due to the limited methodological quality of included RCTs.

Recently, the prevalence of chromosomal abnormalities (CA) increased as the increasing proportion of mothers with advanced age. We aimed to explore the prevalence of CA in relation to maternal age and perinatal features. A retrospective study was performed based on provincial birth defects surveillance data. The relative risk (RR) and 95% confidence interval (CI) were used to calculate maternal age-specific rates of CA. Socio-demographic characteristics of mothers and perinatal features were listed. The total prevalence of CA was 6.38 per 10,000 births, which increased per 10,000 births linearly from 4.02 in 2011 to 9.13 in 2015 (x 2 line-trend =52.69, p

Objectives To explore whether transcutaneous electrical acupoint stimulation (TEAS) can improve the outcomes of in vitro fertilization (IVF). Design A prospective, randomized and controlled study. Setting IVF center in a university hospital. Participants Four hundred and eighty-one infertile patients with bilateral tubal blockage who were referred for IVF. Patients were randomized into four groups. Intervention TEAS was administered for 30 min, respectively at 24 h before TVOR and 2 h before ET. The acupoints included SP10 (Xuehai, bilateral), SP8 (Diji, bilateral), LR3 (Taichong, bilateral), ST36 (Zusanli, bilateral), EX-CA1 (Zigong, bilateral), RN4 (Guanyuan), PC6 (Neiguan, bilateral) and RN12 (Zhongwan). Based on different frequencies of TEAS, patients were grouped into a TEAS-2 Hz group, a TEAS-100 Hz group and a TEAS-2/100 Hz group. Patients in the control group only received routine IVF treatment and no TEAS were applied on them. Primary and secondary outcome measures The number of mature oocytes, normally fertilized oocytes and good-quality embryos were used to evaluate oocyte developmental competence of the patients. Data of clinical pregnancy rate (CPR), implantation rate (IR) and live birth rate (LBR) were also obtained. The levels of neuropeptide Y (NPY), transforming growth factor alpha and granulocyte colony-stimulating factor in the follicular fluids were measured with enzyme-linked immunosorbent assay (ELISA). Results No significant differences were found between the control, TEAS-2 Hz, TEAS-100 Hz and TEAS-2/100 Hz groups on the numbers of metaphase II oocytes, normally fertilized zygotes, early cleavage embryos or good quality embryos (P>0.05). However, the CPR, IR and LBR of the TEAS-2/100 Hz group were significantly higher than those of the other groups, respectively (P

Bicarbonate (HCO3−) is essential for preimplantation embryo development. However, the mechanism underlying the HCO3− transport into the embryo remains elusive. In the present study, we examined the possible involvement of Cl−/HCO3− exchanger in mediating HCO3− transport into the embryo. Our results showed that depletion of extracellular Cl−, even in the presence of HCO3−, suppressed embryo cleavage in a concentration-dependent manner. Cleavage-associated HCO3−-dependent events, including increase of intracellular pH, upregulation of miR-125b and downregulation of p53, also required Cl−. We further showed that Cl−/HCO3− exchanger solute carrier family 26 (SLC26) A3 and A6 were expressed at 2-cell through blastocyst stage. Blocking individual exchanger’s activity by inhibitors or gene knockdown differentially decreased embryo cleavage and inhibited HCO3−-dependent events, while inhibiting/knocking down both produced an additive effect to an extent similar to that observed when CFTR was inhibited. These results indicate the involvement of SLC26A3 and A6 in transporting HCO3− essential for embryo cleavage, possibly working in concert with CFTR through a Cl− recycling pathway. The present study sheds light into our understanding of molecular mechanisms regulating embryo cleavage by the female reproductive tract.

Gametogenesis is a biological process by which diploid or haploid precursor cells undergo cell division and differentiation to form mature haploid gametes. The biology of gamete production is different between males and females. In human, gametogenesis is the development of diploid germ cells into either haploid sperm (spermatogenesis) or eggs (oogenesis). Gamete production in females is intimately part of the endocrine responsibility of the ovary. If there are no gametes, then hormone production is drastically curtailed. Depletion of oocytes implies depletion of the major hormones of the ovary. In the male this is not the case. Androgen production will proceed normally without a single spermatozoon in the testes. This chapter reviews some of the basic structural morphology and physiology of the testes and the ovaries.

Background/Aims: Water channels, also named aquaporins (AQPs), play crucial roles in cellular water homeostasis. Methods: RT-PCR indicated the mRNA expression of AQPs 1-5, 7, 9, and 11-12, but not AQPs 0, 6, 8, and 10 in the 2∼8-cell stage human embryos. AQP3 and AQP7 were further analyzed for their mRNA expression and protein expression in the oocyte, zygote, 2-cell embryo, 4-cell embryo, 8-cell embryo, morula, and blastocyst from both human and mouse using RT-PCR and immunofluorescence, respectively. Results: AQP3 and AQP7 were detected in all these stages. Knockdown of either AQP3 or AQP7 by targeted siRNA injection into 2-cell mouse embryos significantly inhibited preimplantation embryo development. However, knockdown of AQP3 in JAr spheroid did not affect its attachment to Ishikawa cells. Conclusion: These data demonstrate that multiple aquaporins are expressed in the early stage human embryos and that AQP3 and AQP7 may play a role in preimplantation mouse embryo development.

To study the correlation between sperm DNA fragmentation index (DFI), age of male, various parameters of sperm, rates of fertilization, high quality embryo and pregnancy and implantation rates.One hundred and eleven infertile couples were selected randomly, and DFI was tested by flow cytometry for the sperm used for IVF. The patients were divided into different groups according to the DFI scores. The results of each group were analyzed.The IVF normal fertilization was significantly lower in couples with sperm DFI over 10% (60.5%) than that in couples with DFI below 10% (70.1%) (P0.05). Significantly positive correlation was found between DFI and the age of male (r=0.624, P0.05). DFI was also significantly negatively correlated with the percentage of linearly progressive sperm (r=-0.360, P0.05). There was no significant correlation between the rates of high quality cleaved embryos, pregnancy and implantation rate and sperm DFI.DFI scores are increased with male's age, and it can influence the sperm motility. DFI=10% can be considered as a critical point which can be used to estimate the clinical fertility rate of IVF. But it could not provide relative information about the rates of high quality embryos and pregnancy for infertile couples undergoing IVF procedure.

To analyze the numerical aberration rate of X, Y and chromosome 18 in sperms from an oligozoospermic male with mosaic trisomy 18 and to perform preimplantation genetic diagnosis (PGD) for the couple.G-banding and fluorescence in situ hybridization (FISH) were performed on metaphase chromosome. Sperm was analyzed in three-color FISH with a probe mixture containing CEP18, CEPY and Tel Xq/Yq. A healthy man with normal semen parameters was used as control.Significant difference in the rates of disomy for chromosome 18 (0.63% vs. 0.16%) and the gonosomes (0.945% vs. 0.35%) and diploidy (0.87% vs. 0.31%) was found in the spermatozoa between the patient and the control. After four embryos were biopsied in one PGD cycle, two embryos with XY1818 and XX1818 were selected for implanting and clinical pregnancy was ongoing.Sperm-FISH allows further understanding of aneuploidy rate and accurate genetic counseling. FISHPGD was effective for patient with mosaic trisomy 18.

To investigate the influence of different cycles, ovarian follicle size and IVM culture media on the number of retrieved immature oocytes, maturation rate, fertilization rate, embryo quality and implantation rate, pregnancy rate, delivery rate, survival and development of frozen-thawed embryos from IVM.The oocytes were obtained by follicular aspiration from 19 women undergoing oocyte retrieval for in vitro maturation due to the possible risk of ovarian hyperstimulation in IVF-ET program. One patient was in natural cycle, four patients were in ovulation induction cycles with gonadotropine and fourteen patients is controlled ovarian stimulated cycles. All the oocytes retrieved from follicles with 10.0 - 13.5 mm in maximumdiameter were allowed to culture in medium M-199 (TCM 199) or HTF supplemented with other substance.When there were nonuniform diameters of follicles and the diameter of largest oocyte exceeded 12 mm, the retrieval rate of oocytes, fertilization rate, and the number of high-quality embryos decreased. The high-quality embryos formation rate was higher for the oocytes cultured in TCM 199 medium than in HTF medium (P0.01). After being frozen-thawed, the IVM embryos could achieve the same outcome when compared with the conventional IVF treatment. In addition, the offspring were healthy.When the nonuniform diameters of follicles and the diameter of largest oocyte exceeds 12 mm,the retrieval rate of oocytes, fertilization rate, and the number of high-quality embryos decreased. TCM199-based medium is better to improve the developmental potential and implantation rate of embryos derived from in vitro matured oocytes. After being frozen-thawed, the IVM embryos could achieve the same outcome when compared with the conventional IVF treatment. In addition, the offspring are healthy.

To perform preimplantation genetic diagnosis (PGD) of chromosome abnormality using fluorescence in-situ hybridization (FISH).Ten couples were presented for preimplantation genetic diagnosis. They had a total of 10 oocyte pick-up cycles. The collected oocytes were inseminated by intracytoplasmic sperm injection. PGD was carried out using cleavage-stage (day 3) embryo biopsy, fluorescence in-situ hybridization, and day 4 embryo transfer.Ten oocyte pick-up cycles yielded 158 oocytes. Among the 94 embryos obtained, 54 embryos were biopsied and FISH analyses were performed for 51 blastomeres. Twenty-four embryos were transferred on the fourth day. There were 4 clinical pregnancies: 3 infants have been born, and 1 couple had ectopic pregnancy.PGD is a valuable method to prevent the high risk of spontaneous miscarriages and conceiving chromosomally unbalanced offspring.

OBJECTIVE: To explore the influence of polyspermy on IVF outcomes in in vitro fertilization and embryo transfer(IVF-ET). METHODS: The data from 496 IVF-ET cycles and 5349 oocytes were analyzed retrospectively. A comparison of a number of fertility parameters with and without polyspermy was done. The fertility parameters were the number of oocytes retrieved, percentage of mature oocytes, fertilization rate, cleavage rate, occytes for ET, pregnancy rate. RESULTS: The percentage of mature occytes, fertilization rate, cleavage rate was 67.0 %,76.7 %and 95.6 %, respectively( P0.01). The pregnancy rate was higher in polyspermic fertilization cycles (25.7 %) than in cycles without polyspermy(23.6 %),but with no statistical significance ( P0.05). CONCLUSION: Polyspermic fertilization is correlated with improved oocyte receptibility to sperm and could be considered as an encouraging sign for the success of IVF.

OBJECTIVE: To establish a technology of preimplantation genetic diagnosis. METHODS: Intracytoplasm sperm injection and blastomere biopsy were performed on two women at the advanced age with the fallopian tube obstruction. Normal embryos were selected for embryo transfer after fluorescence in-situ hybridziation in biopsied blastomere. RESULTS: The levels of serum HCG were increased 20 days after embryo transfer and ultrasonography in 16 gestation weeks showed the fetal growth and structure are normal. CONCLUSION: Two successful clinical pregnancies achieved after preimplantation genetic diagnosis.