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Protect chromosomes from end-to-end fusion during meiotic bouquet

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Science China(Life Sciences) 2018年第6期61卷 736-738页

作者： Shunxin Wang Binyuan Zhai Xiao Yang Liangran Zhang Center for reproductive Medicine Shandong University Shandong Provincial Hospital Affiliated to Shandong University The Key Laboratory of Reproductive Endocrinology of Ministry of Education National Research Center for Assisted Reproductive Technology and Reproductive Genetics Shandong Provincial Key Laboratory of Reproductive Medicine State key laboratory of Microbial Technology Shandong University

Meiosis generates gametes with half the parental chromosome complements,and thus is crucial for successful sexual reproduction.During meiotic prophase I,a series of specialized events occur to ensure correct paternal and maternal homologous chromosomes(homologs)segregation during anaphase I.They include recognition,alignment,pairing and synapsis of the homologs and meiotic recombination

关键词：染色体端对端伤害熔化保护成熟分裂专业化事物

Noninvasive preimplantation genetic testing in assisted reproductive technology:current state and future perspectives

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Journal of Genetics and Genomics 2020年第12期47卷 723-726页

作者： Jingyi Li Yifeng Liu Yuli Qian Dan Zhang key laboratory of reproductive Genetics(ministry of education)and Department of reproductive endocrinology Women's HospitalZhejiang University School of MedicineHangzhouZhejiang 310006China key laboratory of Women's reproductive Health of Zhejiang Province HangzhouZhejiang 310006China

Invasive genetic screening of pre-implantation embryos via biopsied trophectoderm(TE)cells has been in use for more than 20 years,while its benefits in selecting euploid embryos remain controversial.Recent advances in the ability to process embryonic cell-free DNA(cfDNA)from blastocoel fluid(BF)and spent culture media(SCM)of blastocysts in a manner similar to that of a biopsied TE sample provide a potential alternative holding great promise for obtaining cytogenetic information of the embryos without intrusive biopsy of traditional biopsy-based pre-implantation genetic testing(PGT).Several studies have reported even higher diagnostic accuracy in non-invasive PGT(ni-PGT)than conventional PGT.However,there are still several technical challenges to be overcome before ni-PGT can be accepted as a reliable genomic information source of embryo.In this review,we have summarized the emergence and current state of ni-PGT,and discussed our own perspectives on their limitations and future prospect.There is still a long way to go before truly wide clinical application of ni-PGT.

关键词： Pre-implantation genetic testing Spent culture media Blastocoel fluid Cell-free DNA Non-invasive PGT

Female Fertility： Is it Safe to Freeze.

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Chinese Medical Journal 2015年第3期128卷 390-397页

作者： Lu Zhang Li-Ying Yan Xu Zhi Jie Yah Jie Qiao Department of Obstetrics and Gynecology Center for Reproductive Medicine Peking University Third Hospital Key Laboratory of Assisted Reproduction Ministry of Education Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction Beijing 100191 China

Objective： To evaluate the safety and risk of cryopreservation in female fertility preservation. Data sources： The data analyzed in this review were the English articles from 1980 to 2013 from journal databases, primarily PubMed and Google scholar. The criteria used in the literature search show as tbllowing：（ 1 ） human; embryo; cryopreservation/freezing/vitrification, （2） human; oocyte/immature oocyte; cryopreservation/freezing/vitrification, （3） human; ovarian tissue transplantation; cryopreservation/ freezing/vitrification, （4） human; aneuploidy/DNA damage/epigenetic; cryopreservation/freezing/vitrification, and （5） human; fertility preservation; maternal age. Study selection： The risk ratios based on survival rate, maturation rate, fertilization rate, cleavage rate, implantation rate. pregnancy rate. and clinical risk rate were acquired from relevant meta-analysis studies. These studies included randomized controlled trials or studies with one of tile primary outcome measures covering cryopreservation of human mature oocytes, embryos, and ovarian tissues within the last 7 years （from 2006 to 2013. since the pregnancy rates of oocyte vitrification were significantly increased due to the improved techniques）. The data involving immature oocyte cryopreservation obtained from individual studies was also reviewed by the at, thors. Results： Vitrifications of mature oocytes and embryos obtained better clinical outcomes and did not increase the risks of DNA damage, spindle configuration, embryonic aneuploidy, and genomic imprinting as compared with fresh and slow-freezing procedures, respectively. Conclusions： Both embryo and oocyte vitrifications are safe applications in female fertility preservation.

关键词： Embryo Cryopreservation Oocyte Cryopreservation： Ovarian Tissue Cryopreservation

Where are the theca cells from: the mechanism of theca cells derivation and differentiation

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Chinese Medical Journal 2020年第14期133卷 1711-1718页

作者： Tao Liu Qing-Yuan Qin Jiang-Xue Qu Hai-Yan Wang Jie Yan Department of Obstetrics and Gynecology Center for Reproductive MedicinePeking University Third HospitalBeijing 100191China key laboratory of Assisted Reproduction Ministry of EducationBeijing 100191China Beijing key laboratory of reproductive endocrinology and Assisted Reproduction Beijing 100191China

Mammalian follicles are composed of oocytes,granulosa cells,and theca cells.Theca cells form in the secondary follicles,maintaining follicular structural integrity and secreting steroid hormones.Two main sources of theca cells exist:Wilms tumor 1 positive(Wt1+)cells native to the ovary and Gli1+mesenchymal cells migrated from the mesonephros.Normal folliculogenesis is a process where oocytes,granulosa cells,and theca cells constantly interact with and support each other through autocrine and paracrine mechanisms.The proliferation and differentiation of theca cells are regulated by oocyte-derived factors,including growth development factor 9 and bone morphogenetic protein 15,and granulosa cell-derived factors,including desert hedgehog,Indian hedgehog,kit ligand,insulin-like growth factor 1,as well as hormones such as insulin and growth hormones.Current research on the origin of theca cells is limited.Identifying the origin of theca cells will help us to systematically elaborate the mechanisms of follicular formation and development.

关键词： Theca cell Hedgehog pathway Growth development factor 9 Bone morphogenetic protein 15 TGF-b superfamily Kit ligand

Clinical applications of MARSALA for preimplantation genetic diagnosis of spinal muscular atrophy

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Journal of Genetics and Genomics 2016年第9期43卷 541-547页

作者： Yixin Ren Xu Zhi Xiaohui Zhu Jin Huang Ying Lian Rong Li Hongyan Jin Yan Zhang Wenxin Zhang Yanli Nie Yuan Wei Zhaohui Liu Donghong Song Ping Liu Jie Qiao Liying Yan Department of Obstetrics and Gynecology Third Hospital Peking University key laboratory of Assisted Reproduction Ministry of Education and Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology Peking-Tsinghua Center for Life Sciences Peking University

Conventional PCR methods combined with linkage analysis based on short tandem repeats （STRs） or Karyomapping with single nucleotide polymorphism （SNP） arrays, have been applied to preimplantation genetic diagnosis （PGD） for spinal muscular atrophy （SMA）, an autosome recessive disorder. However, it has limitations in SMA diagnosis by Karyomapping, and these methods are unable to distinguish wild- type embryos with carriers effectively. Mutated allele revealed by sequencing with aneuploidy and linkage analyses （MARSALA） is a new method allowing embryo selection by a one-step next-generation sequencing （NGS） procedure, which has been applied in PGD for both autosome dominant and X-linked diseases in our group previously. In this study, we carried out PGD based on MARSALA for two carrier families with SMA affected children. As a result, one of the couples has given birth to a healthy baby free of mutations in SMA-causing gene. It is the first time that MARSALA was applied to PGD for SMA, and we can distinguish the embryos with heterozygous deletion （carriers） from the wild-type （normal） ones accurately through this NGS-based method. In addition, direct mutation detection allows us to identify the affected embryos （homozygous deletion）, which can be regarded as probands for linkage analysis, in case that the affected family member is absent, In the future, the NGS-based MARSALA method is expected to be used in PGD for all monogenetic disorders with known pathogenic gene mutation.

关键词： Preimplantation genetic diagnosis Spinal muscular atrophy Next-generation sequencing Mutated allele revealed by sequencing with aneuploidy and linkage analyses

Antiphospholipid Antibody Titers and Clinical Outcomes in Patients with Recurrent Miscarriage and Antiphospholipid Antibody Syndrome： A Prospective Study

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Chinese Medical Journal 2017年第3期130卷 267-272页

作者： Yu Song Hai-Yan Wang Jie Qiao Ping Liu Hong-Bin Chi reproductive Medical Center Department of Obstetrics and Gynecology Peking University Third Hospital Beijing 100191 China Beijing key laboratory of reproductive endocrinology and Assisted reproductive Technology Beijing 100191 China key laboratory of Assisted Reproduction ministry of education Beijing 100191 China

Background： The management of patients with recurrent miscarriage （RM） and antiphospholipid antibody syndrome （APS） includes prolonged treatment with heparin and aspirin, starting from the confirmation of pregnancy and continuing until 6 weeks after birth. This study was conducted to determine the relationship between changes in antiphospholipid antibody titers and clinical outcomes. The effect of a shortened treatment regimen was also evaluated. Methods： A prospective study of 123 patients with RM and APS between March 2012 and May 2014 was conducted. Patients were pretreated with a low dose of prednisone plus aspirin before pregnancy, and heparin was added after conception. The levels of antiphospholipid antibodies and pregnancy outcomes were evaluated. Results： All patients were positive for anti-β2-glycoprotein 1 （anti-β2-GP 1） IgM. Atier prepregnancy treatment with low-dose prednisone plus aspirin, 99 of 123 patients became pregnant, and 87 of those pregnancies resulted in successful live births, while 12 resulted in miscarriage, showing a success rate of 87.9%. In the live birth group, levels of anti-β2-GP1 were 56.8±49.0 RU/ml before the pretreatment regimen, 32. 1± 26.0 RU/ml after 2 months of pretreatment, and 24.1 ± 23. IRU/ml during early pregnancy （P 〈 0.05）. In the miscarriage group, antiphospholipid antibody titers were 52.8 ±30.7 RU/ml before pretreatment, 38.5 ±34.2 RU/ml after pretreatment, and 33.9 ±24.7 RU/ml during early pregnancy; the decrease in antiphospholipid antibodies was lower in the miscarriage group than in the live birth group （P 〈 0.05）. Of the 24 inferthe patients, the average antibody titer did not decline after pretreatment （P = 0.802）. Conclusions： Anti-[32-GP1 IgM was the predominant form of antibody in patients with RM and APS. The decreases in antiphospholipid antibody titers correlated with better pregnancy outcomes. The shorter treatment regimen was effective and economical.

关键词： Anticoagulant Therapy Antiphospholipid Antibody Antiphospholipid Antibody Syndrome Recurrent Miscarriage

Different sperm sources and parameters can influence intracytoplasmic sperm injection outcomes before embryo implantation

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2012年第1期13卷 1-10页

作者： Yue-hong LU1,2, Hui-juan GAO1, Bai-jia LI1, Ying-ming ZHENG1, Ying-hui YE1, Yu-li QIAN1, Chen-ming XU1, He-feng HUANG1, Fan JIN1 (1Department of reproductive endocrinology, key laboratory of reproductive Genetics, ministry of education, and key laboratory of Women’s reproductive Health of Zhejiang Province, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China) (2Department of reproductive Health, Shaoxing Women and Children Hospital, Shaoxing 312000, China) Department of reproductive endocrinology Key Laboratory of Reproductive Genetics Ministry of Education Women's Hospital School of Medicine Zhejiang University Hangzhou 310006 China.

To evaluate the effects of sperm with different parameters and sources on the outcomes of intracytoplasmic sperm injection (ICSI), 1972 ICSI cycles were analyzed retrospectively. Groups 1 to 5 were composed of cycles using ejaculated sperm and were grouped according to sperm quantity, quality, and morphology into normal (288 cycles), or mild (329 cycles), moderate (522 cycles), severe (332 cycles), and extremely severe (171 cycles) oligozoospermia and/or asthenozoospermia and/or teratozoospermia (OAT) groups. Group 6 was composed of 250 cycles using testicular or epididymal sperm, and Group 7 consisted of 80 cycles using frozen-thawed sperm. We found that fertilization rates were gradually reduced from Groups 1 to 6, and reached statistical difference in Groups 5 and 6 (P0.05). A total of nine cases of malformation were observed, with a malformation rate of 1.25% (9/719). In conclusion, different sperm sources and parameters can affect ICSI outcomes before embryo implantation. A full assessment of offspring mal-formation will require further study using a larger sample size.

关键词：精子数量胚胎植入注射低出生体重使用周期胞浆 ICSI 畸形精子

Elective single blastocyst transfer is more suitable for normal responders than for high responders

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Chinese Medical Journal 2013年第11期126卷 2125-2128页

作者： WU Ke-liang ZHAO Hai-bin LIU Hui ZHONG Wan-xia YU Guan-ling CHEN Zi-jiang Center for reproductive Medicine Provincial Hospital Affiliated to Shandong University National Research Center for Assisted Reproductive Technology and Reproductive Genetics China Key laboratory for Reproductive Endocrinology of Ministry of Education Shandong Provincial Key Laboratory of Reproductive Medicine Jinan Shandong 250021 China

Background Embryo quality and receptivity of the endometrium are two factors that determine the results of in vitro fertilization/intra-cytoplasmic sperm injection-embryo transfer （IVF/ICSI-ET）. There is no consensus of the optimal transfer strategy for normal responders or high responders. The current study aimed to find the optimal transfer strategy for different subgroups of patients. Methods From April 2010 to December 2010, patients who meet the following criteria were included in this study; primary infertility, female age 〈35 years, FSH level on female cycle day 2-3 〈12 mlU/ml, at least six good quality embryos available on day three.The clinical outcomes using different transfer strategies between normal responders and high responders were reviewed and compared. Results For the normal responders, the clinical pregnancy rate of day three double-embryo transfer （DET） was comparable to that of day five elective single blastocyst transfer （eSBT）, 64.04% vs. 60.33% （P〉0.05）. For the high responders, the clinical pregnancy rate of day five eSBT was significantly lower than that of day three DET, 43.35% vs. 57.21% （P〈0.05）. For the high responders, the rates of clinical pregnancy and implantation in frozen-thawed embryo transfer （FET） cycles were notably higher than in eSBT cycles （64.56% vs. 43.35% and 62.11% vs. 43.35% respectively）（P〈0.05）. Conclusions For normal responders, eSBT might be an applicabte strategy to reduce multiple pregnancy rates while maintaining acceptable overall pregnancy rates. And in order to reduce multiple pregnancies and increase the chance of pregnancy of high responders, FET may be a preferable strategy.

关键词： embryo transfer elective single blastocyst transfer high responder normal responder

Effect of vitrification at the germinal vesicle stage on the global methylation status in mouse oocytes subsequently matured in vitro

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Chinese Medical Journal 2014年第23期127卷 4019-4024页

作者： Yan Jie Zhang Lu Wang Tianren Li Rong Liu Ping Yan Liying Qiao Jie Department of Obstetrics and Gynecology Peking University Third Hospital Key Laboratory of Assisted Reproduction Ministry of Education Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction Beijing 100191 China

Background It is still unclear whether the vitrification procedure itself is associated with the incidence of abnormal DNA methylation during oocytes vitrification.The purpose of this study was to evaluate the epigenetic profile of mouse oocytes,which went through vitrification either at a mature stage or at an immature stage following in vitro maturation （IVM） by analyzing the global DNA methylation.Methods Metaphase Ⅱ （M Ⅱ） stage and germinal vesicle （GV） stage oocytes were collected from adult female mice and were vitrified respectively.The M Ⅱ oocytes were assessed for cryo-survival and global DNA methylation.The GV oocytes were assessed for cryo-survival and only the surviving GV oocytes were cultured in vitro for subsequent assessment of global DNA methylation in mature oocytes.In vivo matured fresh M Ⅱ oocytes without undergoing vitrification were used as control.The level of global DNA methylation in the M Ⅱ oocytes was then examined by immunofluorescence using an anti-5-methylcytosine （anti-5-MeC） monoclonal antibody and fluorescein isothiocyanate （FITC）-conjugated goat anti-mouse IgG under a laser scanning confocal microscope.Results In terms of the effect of vitrification on global DNA methylation status in matured oocytes,in the M Ⅱ-v group,all the examined oocytes （90/90） were found with hypermethylation,including 63.3％（57/90） of them displaying DNA methylation of a very high level,25.6％（23/90） with a high level,and 11.1％（10/90） with an intermediate level,whereas in the GV-v group,all the matured oocytes （129/129） were also examined with hypermethylation,including 67.4％（87/129） of them displaying DNA methylation of a very high level,23.3％（30/129） with a high level,and 9.3％（12/129） with an intermediate level.Statistically,it was similar between both groups,which were similar to the control：68.6％（83/121） of fresh M Ⅱ oocytes displayed DNA methylation of a very high level,21.5％（26/121） with a high level,and 9.9％（12/121） with an intermediate level （P ＞0.05）.In terms of the effect of IVM on global

关键词： DNA methylation in vitro maturation oocyte vitrification epigenetic safety