Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

Fertilization is a crucial step for origin of life.During Assisted Reproductive Technologies (ART),total fertilization failure is complex and unpredictable.Total fertilization failure may related to some abnormal cellular mechanistic events,such as:any stage of sperm and cumulus-oocyte-complexes penetration,sperm-zona pellucida binding / penetration,sperm-oocyte membrane binding,oocyte activation,sperm discondensation or pronuclear formation.Most of total fertilization failure could be solved by intracytoplasmic sperm injection.But oocytes of some patient still can't fertilize successfully,even though assisted oocyte activation be used.As for total fertilization failure patients in ART,combining the mature of oocyte,sperm quality and some trail to improve clinical protocol in later cycle may prevent failure to happen again.

OBJECTIVE: To explore the effect of calcitonin gene-related peptide (CGRP) on murine oocyte maturation. METHODS: After injection of pregnant mare serum gonadotropin (PMSG, 10 U, i.p.) for 48 h, 6-week old female Kunming mice were killed, and the cumulus oocyte complexes (COCs) were collected from ovaries and inoculated in the culture plate by 30-40/hole. The COCs were treated with 4 concentrations of CGRP (0, 10(-10), 10(-9), and 10(-8) mol/L), and the germinal vesicle breakdown (GVBD) and polar body I (PBI) were examined. Human granulosa cells were also cultured with CGRP (0, 10(-10), 10(-9), 10(-8) mol/L) and levels of intracellular cyclic adenosine monophosphate (cAMP) were measured. RESULTS: Exogenous CGRP caused a decrease in GVBD and PBI in COCs, and an increase in cAMP levels in human granulosa cells in a concentration-dependent manner. CONCLUSION CGRP can inhibit the oocyte maturation, which may be related to the increased content of cAMP in granulosa cells.
Animals ; Calcitonin Gene-Related Peptide ; pharmacology ; Cyclic AMP ; metabolism ; Female ; Granulosa Cells ; cytology ; Humans ; In Vitro Techniques ; Mice ; Oocytes ; cytology ; drug effects

Objective To compare the therapeutic effect of two Methodsof in vitro oocyte maturation (IVM) technology with polycystic ovary syndrome (PCOS) patients in infertility treatment.MethodsRetrospective analysis was performed on patients with PCOS infertility who were stimulated with two different kinds of stimulating method of the IVM cycle in our center during April 2010 and August 2010.There were 20 patients in the temporary FSH stimulating group (group A) and 31 patients in the HCG stimulating group (group B).The number of oocytes maturation rate,fertilization rate,cleavage rate,excellent embryo rate,implantation rate,pregnancy rate per oocyte recycle,pregnancy rate per transfer and abortion rate of transfer were compared.ResultsThere was no significant difference between the two groups of age,duration of infertility,body mass index,based sex hormone levels by t test(P＞0.05).There was no significant difference between the two groups of the number of oocytes retrieved,oocyte maturation rate,cleavage rate,and abortion rate of transfer(P＞0.05).In group B,the fertilization rate was 80.3%,good-quality-embryo rate was 49.8%,implantation rate was 31.5%,pregnancy rate per oocyte recycle was 54.8%,which was significantly higher than in group A (73.9%,35.4%,12.5% and 25.0%,respectively).ConclusionsIf IVM was used to treat PCOS patients with infertility,HCG stimulating will be better than FSH stimulating,and it could result in a higher clinical pregnancy rate.

Objective To develop a method for treating infertile women with polycystic ovary syndrome(PCOS)by ooeyte in vitro maturation(IVM).Methods On day 3～4 of menstrual cycle,infertile women with PCOS were administrated with(100～150IU)FSH/day for 3 days.On day 8～10,when the follicle diameter reached 10～14mm and endometrium＞5mm,the immature oocytes weFe picked up and cultured in vitro for 28～31 hours.Then,ICSI and embryo transfer were performed.Results 3 clinic pregnancies were established among 12 infertile women.the pregnancy rate was 25.0%.Conclusion This research indicates that IVM is a feasible technology for the infertile women with PCOS.