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Journal of reproduction and Contraception 2014年第2期25卷 81-88页

作者： Jian-ping ZHANG Xue-qing WU Xing-yu BI Bing-bing CHEN Si-ping ZHANG Hui-ping LIU Human assisted reproduction center Shanxi Women & Children's Hospital

Objective To investigate the effect of different down-regulation protocol on the in vitro fertilization-embryo transfer （IVF-ET） outcomes in infertile patients with endometriosis （EMs）.Methods A retrospective case control study was performed. Totally 294 infertile patients with EMs were enrolled. And 109 patients （116 cycles） received prolongedprotocol as the control, 185 patients （193 cycles） received long protocol as case group, all followed by standard controlled ovarian hyperstimulation （COH）. Response to gonadotropins, the fertilization rate, the cleavage rate, the implantation rate, the clinical pregnancy rate and the miscarriage rate were measured and analyzed between the two groups. Results A trend toward better ovarian response was observed in long protocol group. Higher fertilization rate, lower total dose of rFSH, shorter duration of stimulation and more endometrial thickness on the day of hCG injection were observed in long protocol group compared with those of prolonged protocol group, and the difference was significant （P〈0.05）. In addition, the clinical pregnancy rate, the cleavage rate and the implantation rate also had an increase trend in long protocol group compared with those of prolonged protocol group, but without significant differences. Conclusion Long protocol regimen before IVF-ET in patients with EMs resulted in a trend toward better ovarian response and higher clinical pregnancy rates than prolonged protocol regimen did.

关键词： endometriosis (EMs) down-regulation protocol controlled ovarianhyperstimulation (COH) IVF-ET pregnancy rate

Sperm origins and concentration do not impact the clinical outcomes in intracytoplasmic sperm injection cycles

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Asian Journal of Andrology 2018年第5期20卷 454-458页

作者： Cen Yang Ze-Hong Zhou Dan-Ni Zheng Xiao-Fei XU Jin Huang Ying Lian Jie Qiao center for Reproductive Medicine Department of Obstetrics and Gynecology Peking University Third Hospital Beijing 100191 China Beijing Key Laboratory of Reproductive Endocrinology and assisted reproduction Technology Beijing 100191 China Key Laboratory of assisted reproduction Ministry Of Education Beijing 100191 China

In the present study, we evaluated the impact of sperm origins and concentration on the clinical outcomes of intracytoplasmic sperm injection （ICSI） cycles. A total of 1201 IC51 cycles were retrospectively analyzed for male azoospermia or oligozoospermia between January 2015 and December 2015 in the Peking University Third Hospital. Patients were divided into three groups （Group 1 vs Group 2/3; surgically extracted sperm vs ejaculated sperms）： Group 1 included 343 ICSI cycles and Group 2 analyzed 388 cycles on semen with sperm concentration 〈5 × 10^6 m1-1 （severe oligozoospermia group）. Group 3 included 470 cycles with sperm concentration between 5×10^6 m1-1 and 15 × 10^6 m1-1 （mild oligozoospermia group）. Fertilization rates, clinical pregnancy rates, and live birth rates were analyzed and compared among groups of different semen origins and concentrations on the oocyte retrieval day. Group 2 showed a lower fertilization rate than Group 3 （62.9%± 21.6% vs 66.8% ± 22.1%, P 〈 0.05）. There were no statistically significant differences in clinical pregnancy rate per transfer （51.3%, 46.7%, and 50.0%, respectively）, live birth rate per transfer （44.4%, 40.9%, and 41.4%, respectively）, accumulative live birth rate （58.3%, 51.0%, and 52.1%, respectively）, twin birth rate （18.4%, 10.6%, and 12.6%, respectively）, and birth defects rate （0, 0.3%, and 0.2%, respectively） among three groups. The results of this study indicated that sperm origins and concentration do not imDact the clinical outcomes in ICSI cycles.

关键词： azoospermia clinical outcomes intracytoplasmic sperm injection oligozoospermia sperm concentration

AMP-activated kinase in human spermatozoa： identification, intracellular localization, and key function in the reRulation of sperm motility

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Asian Journal of Andrology 2017年第6期19卷 707-714页

作者： Violeta Calle-Guisado Ana Hurtado de Llera David Martin-Hidalgo Jose Mijares Maria C Gil Ignacio S Alvarez Maria J Bragado Luis J Garcia-Marin Research Group of Intracellular Signalling and Technology of reproduction (SINTREP) School of Veterinary Medicine University of Extremadura Caceres Spain assisted reproduction Unit at the-Minimally Invasive surgery center Jesos Uson (CCMIJU) Caceres Spain Norba Clinic Caceres Spain Research Group of reproduction and Embryo Development (REDES) University of Extremadura Badajoz Spain Extremadura Institute of assisted reproduction (IERA) Badajoz Spain.

AMP-activated kinase （AMPK）, a protein that regulates energy balance and metabolism, has recently been identified in boar spermatozoa where regulates key functional sperm processes essential for fertilization. This work＇s aims are AMPK identification, intracellular localization, and their role in human spermatozoa function. Semen was obtained from healthy human donors. Sperm AMPK and phospho-Thr172-AMPK were analyzed by Western blotting and indirect immunofluorescence. High- and low-quality sperm populations were separated by a 40%-80% density gradient. Human spermatozoa motility was evaluated by an Integrated Semen Analysis System （ISAS） in the presence or absence of the AMPK inhibitor compound C （CC）. AMPK is localized along the human spermatozoa, at the entire acrosome, midpiece and tail with variable intensity, whereas its active form, phospho-Thr172-AMPK, shows a prominent staining at the acrosome and sperm tail with a weaker staining in the midpiece and the postacrosomal region. Interestingly, spermatozoa bearing an excess residual cytoplasm show strong AMPK staining in this subcellular compartment. Both AMPK and phospho-Thr172-AMPK human spermatozoa contents exhibit important individual variations. Moreover, active AMPK is predominant in the high motility sperm population, where shows a stronger intensity compared with the low motility sperm population. Inhibition of AMPK activity in human spermatozoa by CC treatment leads to a significant reduction in any sperm motility parameter analyzed： percent of motile sperm, sperm velocities, progressivity, and other motility coefficients. This work identifies and points out AMPK as a new molecular mechanism involved in human spermatozoa motility. Further AMPK implications in the clinical efficiency of assisted reproduction and in other reproductive areas need to be studied.

关键词： AMP-activated kinase human spermatozoa immunolocalization sperm motility sperm quality

Effects of malignancies on fertility preservation outcomes and relevant cryobiological advances

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Science China(Life Sciences) 2020年第2期63卷 217-227页

作者： Dongming Liu Jie Yan Jie Qiao Reproductive Medical center Department of Obstetrics and GynecologyPeking University Third HospitalBeijing 100191China National Clinical center for Obstetrics and Gynecology Beijing 100191China Key Laboratory of assisted reproduction Ministry of EducationBeijing 100191China Beijing Key Laboratory of Reproductive Endocrinology and assisted reproduction Beijing 100191China

A decrease in cancer deaths has resulted in the possibility of child bearing for many young adult cancer survivors. Most antitumor treatment modalities are detrimental to female fertility, and methods for fertility preservation before gonadotoxic treatment, including cryopreservation of oocytes, embryos and ovarian tissue, have therefore been developed. This review focuses on the ovarian function of cancer patients, the safety and efficacy of fertility preservation methods, and the pregnancy outcomes of these patients. Breast cancer and hematological tumors constitute the majority of cancers in reproductive-aged female oncology patients. Ovarian function may not be impacted by breast cancer cells, while in patients with hematological malignancies, decreases in anti-Müllerian hormone and antral follicle counts have been demonstrated. In most cases, patients can undergo ovarian stimulation without delaying treatment, and a new stimulation protocol known as dual stimulation, which may be more efficient, has now been developed. Birth outcomes are also acceptable in cancer patients.

关键词： cancer patients fertility preservation controlled ovarian stimulation safety

Comparative Study of Fluorescence in situ Hybridization Analysis and Karyotype Analysis in Spontaneous Abortion Etiology Diagnosis

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Journal of reproduction and Contraception 2013年第3期24卷 133-142页

作者： Liang CHANG Su ZHONG Nan ZHAO Ping LIU Jie QIAO center for Reproductive Medicine Department of Obstetrics and GynecologyPeking University Third HospitalKey Laboratory of Assisted ReproductionMinistry of EducationKey Laboratory of Reproductive Endocrinology and Assisted Reproduction

Objective To compare the roles of fluorescence in situ hybridization （FISH） and karyotype analysis on chorionic villus in spontaneous abortion. Methods A total of 201 cases were included in this study and were randomly divided into 2 groups by the same gestational age. The villi from 100 cases （group A） were cultured and used for karyotype analysis, while the uncultured villi from 101 women （group B） were used for FISH. A case was randomly selected from pregnant patients of 6-11 weeks at each gestational week in each group, 6 cases from each group and a total of 12 cases were analyzed by FISH and karyotype at the same time. Results The successful karyotype analysis rate was 66. 0%, and the abnormality karyotype rate was 30.3%; FISH success analysis rate was 100%, and the abnormality rate was 46. 5%; there were significant differences between FISH and karyotype analysis （P=0.036）. Conclusion There were obvious differences between the two techniques. FISH was more successfully analyzed, and was used to more precisely determine fetal chromo- some number. It is an effective way to determine abnormal chromosome by integrating FISH and karyotype analysis in spontaneous abortion.

关键词： spontaneous abortion early pregnancy fluorescence in situ hybridization(FISH) karyotype analysis

Dual role for the unfolded protein response in the ovary： adaption and apoptosis

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Protein & Cell 2017年第1期8卷 14-24页

作者： Ning Huang Yang Yu Jie Qiao Beijing Key Laboratory of Reproductive Endocrinology and assisted Reproductive Technology and Key Laboratory of assisted reproduction Ministry of Education Center of Reproductive Medicine Department of Obstetrics and Gynecology Peking University Third Hospital Beijing 100191 China

The endoplasmic reticulum （ER） is the principal organelle responsible for several specific cellular functions including synthesis and folding of secretory or membrane proteins, lipid metabolism, and Ca^2＋ storage. Different physiological as well as pathological stress conditions can, however, perturb ER homeostasis, giving rise to an accumulation of unfolded or misfolded proteins in the ER lumen, a condition termed ER stress. To deal with an increased folding demand, cells activate the unfolded protein response （UPR）, which is initially protective but can become detrimental if ER stress is severe and prolonged. Accumulating evidence demonstrates a link between the UPR and ovarian development and function, including follicular growth and maturation, follicular atresia, and corpus luteum biogenesis. Additionally, ER stress and the UPR may also play an important role in the ovary under pathological conditions. Understanding the molecular mechanisms related to the dual role of unfolded protein response in the ovarian physiology and pathology may reveal the pathogenesis of some reproductive endocrine diseases and provide a new guidance to improve the assisted reproductive technology. Here we review the current literature and discuss concepts and progress in understanding the UPR, and we also analyze the role of ER stress and the UPR in the ovary.

关键词： endoplasmic reticulum stress obesity,ovary follicle ovarian diseases

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

Multiple factors affecting surgical outcomes and patency rates in use of single-armed two-suture microsurgical vasoepididymostomy： a single surgeon＇s experience with 81 patients

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Asian Journal of Andrology 2016年第1期18卷 129-133页

作者： Kai Hong Lian-Ming Zhao Shi-Xing Xu Wen-Hao Tang Jia-Ming Mao De-Feng Liu Hui Jiang Lu-Lin Ma Jie Qiao Department of Urology Peking University Third Hospital Beijing China center for Reproductive Medicine Department of Obstetrics and Gynecology Peking University Third Hospital Beijing China Key Laboratory of assisted reproduction Ministry of Education Beijing China Beijing Key Laboratory of Reproductive Endocrinology and assisted reproduction Beijing China

Vasoepididymostomy （VE）, as the most challenging procedure in microsurgeries, is often carried out with a double-armed two-suture technique. In this study, we evaluated the efficacy and safety of the single-armed two-suture VEs on humans and studied the factors that could possibly affect the patency rates. From July 2012 to July 2013, we reviewed 81 patients with consecutive primary epididymal obstruction who underwent single-armed two-suture longitudinal intussusception microsurgical VEs by a single surgeon, Kai Hong （KH）. At the same time, we analyzed seven factors that possibly related to the patency rates. With the single-armed technique, a total of 81 men underwent the microsurgical VEs. Data on 62 patients were completely recorded. 19 patients were lost to follow-up. Mean age was 31 years old. Mean follow-up time was 8.8 （2-17） months. The patency rate was 66.1% （41/62）. Natural pregnancy rate was 34.1% （14/41）. Overall pregnancy rate was 22.6% （14/62）. No severe surgical complications were noted. With logistic regression test analysis, there were two factors related to a higher patency rate： anastomosis sites （P = 0.035） and motile sperm found in the epididymal fluid （P = 0.006）. Motile sperm found in the epididymal fluid were associated with a higher patency rate （OR = 11.80, 95% CI = 1.79, 77.65）. The single-armed two-suture longitudinal VE technique is feasible for microsurgical practice. The patency and pregnancy rates are comparable to the doubled-armed technique. Anastomosis sites and motile sperm found in the epididymal fluid were the most two important factors related to higher patency.

关键词： male infertility microsurgery obstructive azoospermia vasoepididymostomy

The present and future of whole-exome sequencing in studying andtreating human reproductive disorders

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Journal of Genetics and Genomics 2018年第10期45卷 517-525页

作者： Wei Guo Xiaohui Zhu Liying Yan Jie Qiao center for Reproductive Medicine Department of Obstetrics and Gynecology Peking University Third Hospital Beijing 100191 China Key Laboratory of assisted reproduction Ministry of Education Beijing 100191 China Beijing Key Laboratory of Reproductive Endocrinology and assisted reproduction Beijing 100191 China Beijing Advanced Innovation center for Cenomics (ICG) Peking University Beijing 100871 China Peking-Tsinghua center for Life Sciences Peking University Beijing 100871 china

The causes of recurrent spontaneous abortion (RSA) and fetal malformations are multifactorial and unclear in most cases. Environmental, maternal, and genetic factors have been shown to contribute to these defects. Whole-exome sequencing (WES) is widely used to detect genetic variations associated with human diseases and has recently been successfully applied to unveil genetic causes of unexplained recurrent spontaneous abortion (URSA) and fetal malformations. Here, we review the current discovery and diagnosis strategies to identify the underlying pathogenic mutations of URSA and fetal malformations using WES technology and propose to further develop WES, both to advance our understanding of these diseases and to eventually lead to targeted therapies for reproductive disorders.

关键词： Whole exome sequencing (WES) Unexplained recurrent spontaneous abortion (URSA) Fetal malformations Human reproduction

Genetic testing and PGD for unexplained recurrent fetal malformations with MAGEL2 gene mutation

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Science China(Life Sciences) 2019年第7期62卷 886-894页

作者： Wei Guo Yanli Nie Zhiqiang Yan Xiaohui Zhu Yuqian Wang Shuo Guan Ying Kuo Wenxin Zhang Xu Zhi Yuan Wei Liying Yan Jie Qiao center for Reproductive Medicine Department of Obstetrics and GynecologyPeking University Third HospitalBeijing 100191China Key Laboratory of assisted reproduction Ministry of EducationBeijing 100191China Beijing Key Laboratory of Reproductive Endocrinology and assisted reproduction Beijing 100191China Bering Advanced Innovation center for Genomics Peking UniversityBeijing 100871China Peking-Tsinghua center for Life Sciences Peking UniversityBeijing 100871China

Birth defects are caused by multiple factors,such as chromosome abnormality,environmental factors,and maternal *** this study,we focused on exploring the genetic causes of a non-consanguineous couple who suffered from four times of unsuccessful pregnancy due to unexplained recurrent fetal malformations with similar symptoms and normal chromosome copy number *** trio-whole exome sequencing(trio-WES) for this couple and one of the affected fetuses,we found a mutation,c.1996 delC on the maternal imprinted gene MAGEL2 that was carried by the affected fetus and husband,leading to Schaaf-Yang *** screen this mutation,we further performed preimplantation genetic diagnosis(PGD) strategy followed by a gene pedigree validation and pathogenicity *** the transfer of a PGD-screened embryo,a normal newborn without previous abnormal symptoms was born(February 15,2019).We present the first data that identified a pathogenic gene(MAGEL2 c.1996 delC) in a fetus with Schaaf-Yang syndrome in the EAS(East Asian) database and overcame this genetic defect by using processed PGD for this couple based on the WES results.

关键词： unexplained recurrent fetal malformations whole exome sequencing (WES) preimplantation genetic diagnosis (PGD) Schaaf-Yang syndrome