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Acetyl-L-carnitine:An Effective Antioxidant against Cryo-damage on Human Spermatozoa with Asthenospermia

Journal of Huazhong University of Science and Technology(Medical Sciences) 2017年第6期37卷 915-921页

作者：邹宇洁杨菁常硕徐望明尹太郎龙文 reproductive Medicine Center Renmin Hospital of Wuhan UniversityWuhan 430060China

A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen（ROS） level after freezing-thawing process. The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia. The cryopreservation of human spermatozoa treated with acetyl-L-carnitine at different concentrations（group B： 2.5 mmol/L, group C： 7.5 mmol/L, group D： 15 mmol/L） was compared with control（group A： no acetyl-L-carnitine given）. For the frozen-thawed spermatozoa, the viability, motility and DNA integrity were measured by comet assay, acrosome integrity by FITC-PNA staining and ROS level was determined in each group. The results showed that there were no significant differences in motility and viability between group A and group B, while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A. As compared with group A, the values for DNA integrity parameters including comet rate（CR）, tail DNA percentage（TD）, tail length（TL） and Oliver tail moment（OTM） were significantly reduced in group C and group D. group C and group D also displayed a higher proportion of intact acrosome than group A. No significant difference in ROS level was found between group A and group B, while with the increase in acetyl-L-carnitine concentration, the ROS level in groups C and D was significantly reduced as compared with that in group A. In conclusion, acetyl-L-carnitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.

关键词： acetyl-L-carnitine human spermatozoa DNA damage acrosome integrity

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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

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Preliminary Evaluation of Safety of Conditionally Replication Adenovirus M4

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2012年第6期32卷 893-898页

作者：陈彩虹方海燕饶玉梅吴鹏何杨马丁高庆蕾 Cancer biology Research Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Center of reproductive Medicine the First Affiliated Hospital of Zhengzhou University Department of Obstetrics and Gynecology the First Affiliated Hospital of Zhengzhou University

Conditionally replication adenovirus M4, which was constructed in our lab, was proved to have good clinical application prospect for its good antitumor and antimetastasis effect. However, clinically applying M4 faces many problems. One of the most important is the safety of M4. In this study, we investigated the safety of M4 by comparing with Adv-TK, which was proved to be safe in Ⅰ–Ⅲ phase clinical trials. M4 and Adv-TK were injected into mice via the tail vein separately, and the mice were sacrificed at the indicated time. Blood was collected for biochemical tests, the liver was harvested for hematoxylin and eosin (H&E) staining and viral quantification, and splenic lymphocytes were separated for adenovirus specific cellular immune response. Our results showed that M4 had no obvious effect on mouse general symptoms. A transient reversible infiltration of inflammatory cells in collect abbacy was only observed in M4 group, and a transient slight increase in Cr level was detected both after M4 and Adv-TK injection. The adenovirus specific cellular immune response induced by M4 was similar to that by Adv-TK, and the distribution and metabolism of M4 in the mouse liver were also similar to those of Adv-TK. It was concluded that conditionally replication adenovirus M4 had the same safety as Adv-TK. The study provides safety basis for the coming clinical trials of M4.

关键词： conditionally replication adenovirus M4 safety blood biochemistry distribution

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Flexible GnRH Antagonist Protocol versus Progestin-primed Ovarian Stimulation (PPOS) Protocol in Patients with Polycystic Ovary Syndrome: Comparison of Clinical Outcomes and Ovarian Response

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Current Medical Science 2019年第3期39卷 431-436页

作者： Zhuo-ni XIAO Jia-li PENG Jing YANG Wang-ming XU Center for reproductive Medicine Renmin Hospital of Wuhan University Wuhan 430060China

Polycystic ovary syndrome (PCOS) is one of the most common causes of infertility in women. Progestin-primed ovarian stimulation (PPOS) protocol, which used oral progestin to prevent premature luteinizing hormone (LH) surges in ovarian stimulation, has been proved to be effective and safe in patients with PCOS. The aim of the present study was to compare the efficacy of PPOS protocol with that of the traditional gonadotropin-releasing hormone (GnRH) antagonist protocol in patients with PCOS. A total of 157 patients undergoing in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) were recruited into this study. The patients were divided into two groups by the stimulation protocols: the GnRH antagonist protocol group and the PPOS protocol group. There was no significant difference in the clinical characteristics between the two groups. Dose and duration of gonadotropin were higher in the PPOS protocol group. Estradiol levels on the day of human chorionic gonadotropin (hCG) administration were significantly lower in the PPOS protocol group. Fertilization rates and the number of good quality embryos were similar between the two groups. Remarkably, we found 6 patients with moderate ovarian hyperstimulation syndrome (OHSS) in the GnRH antagonist protocol group but 0 in the PPOS protocol group. A total of 127 women completed their frozen embryo transfer (FET) cycles. There were no significant differences between the two groups in terms of clinical pregnancy rate per transfer, implantation rate, first-trimester miscarriage rate and on-going pregnancy rate per transfer. To conclude, PPOS protocol decreased the incidence of OHSS without adversely affecting clinical outcomes in patients with PCOS.

关键词： polycystic ovary syndrome ovarian hyperstimulation syndrome progestin-primed ovarian stimulation GnRH antagonist protocol controlled ovarian hyperstimulation

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Impact of Periconceptional Multi-micronutrient Supplementation on Gestation:A Population-based Study

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Biomedical and Environmental Sciences 2013年第1期26卷 23-31页

作者： WANG Ya Fei PEI Li Jun SONG Xin Ming CHEN Gong ZHENG Xiao Ying Institute of Population Research/WHO Collaborating Center on reproductive Health and Population Science Peking University

Objective To examine the effect of periconceptional multi‐micronutrient supplementation on gestation and birth *** A population‐based community intervention program was conducted in 18 counties in *** were divided into an intervention group,who received multi‐micronutrient supplementation from at least 3 months before pregnancy throughout the first trimester,and a control *** women were followed up to record information about birth *** socio‐economic characteristics and main birth outcomes were *** age was further analyzed using survival analysis,to determine the time distribution of *** Periconceptional multi‐micronutrient supplementation was associated with higher birth weight,birth length and occipitofrontal head circumference,and with lower incidence rates for stillbirth,low birth weight,and preterm ***,periconceptional multi‐micronutrient supplementation changed the time distribution of delivery,making the deliveries more clustered in the period between day 275 and day 295 of *** Our study shows that periconceptional multi‐micronutrient supplementation is beneficial for fetal development and optimizes all measured aspects of health in neonates in socioeconomically disadvantaged areas in *** change in time distribution of deliveries caused by multi‐micronutrient supplementation needs further clarification.

关键词： Multi‐micronutrient Supplementation Gestation Birth weight Gestational age

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Effects of MⅡ stage oocytes zona pellucida birefringence on pregnancy outcome

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Asian Pacific Journal of Tropical Medicine 2013年第7期6卷 578-582页

作者： Jia Luo Yan-Wen Xu Ming-Fang Zhang Ling Gao Cong Fang Can-Quan Zhou reproductive Center of the First Affiliated Hospital of Sun Yat-sen University

Objective:To explore the effects of different MⅡstage oocytes zona pellucida birefringence on pregnancy ***:A total of 46 couples with infertile which induced by single cause received in-vitro fertilization treatment were analyzed retrospectively,and randomly divided into the high zona birefringence(HZB)/HZB ***/low zona birefringeuce(LZB) group and LZB/LZB group according to different oocytes zona pellucida *** sperm injection outcome was analyzed and ***:The proportion of HZB oocytes, implantation rate and the pregnancy rate were decreased in three groups(HZB/HZB group>HZB/ LZB group>LZB/LZB group)(Pgroups(P>0.05).Logistic regression analysis showed that factors allecl MⅡstage oocytes zona pellucida birefringence were *** FSH level and the LH level on the day of HCG *** and KSH levels were negatively correlated with the single oocyte zona pellucida birefringence:While the LH level on the day of hCC injection was positively correlated with the single oocylc zona pellucida ***:The primary influence factors on MⅡstage oocytes zona pellucida are *** FSH level and the LH level on the day of hCG *** birefringence value of zona pellucida can affect the pregnancy outcome.

关键词： Oocytes Zona pellucida Intracytoplasmic sperm injection Implantation Pregnancy

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Fecal microbiota transplantation from young donor mice improves ovarian function in aged mice

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Journal of Genetics and Genomics 2022年第11期49卷 1042-1052页

作者： Li Xu Qiankun Zhang Xiaowei Dou Yipeng Wang Jianwei Wang Yong Zhou Xingyin Liu Jing Li State Key Laboratory of reproductive Medicine Nanjing Medical UniversityNanjingJiangsu 211166China Department of Pathogen biology-Microbiology Division State Key Laboratory of Reproductive MedicineNanjing Medical UniversityNanjingJiangsu 211166China Shanghai Personal Biotechnology Shanghai 200231China Department of Obstetrics and Gynecology The Second Affiliated Hospital of Nanjing Medical UniversityNanjingJiangsu 210003China Institute of Pediatric Research Children's Hospital of Soochow UniversitySuzhouJiangsu 215003China Fertility Preservation Laboratory Reproductive Medicine CenterGuangdong Second Provincial General HospitalGuangzhouGuangdong 510317China

Advanced maternal age is characterized by declines in the quantity and quality of oocytes in the ovaries,and the aging process is accompanied by changes in gut microbiota ***,little is known about the relationship between gut microbiota and ovarian *** using fecal microbiota transplantation(FMT)to transplant material from young(5-week-old)into aged(42-week-old)mice,we find that the composition of gut microbiota in FMT-treated mice presents a“younger-like phenotype”and an increase of commensal bacteria,such as Bifidobacterium and ***,the FMT-treated mice show increased anti-inflammatory cytokine IL-4 and decreased pro-inflammatory cytokine IFN-γ.Fertility tests for assessing ovarian function reveal that the first litter size of female FMT-treated mice is significantly higher than that of the non-FMT *** analysis demonstrates a dramatic decrease in follicle atresia and apoptosis as well as an increase in cellular proliferation in the ovaries of the FMT-treated *** results also show that FMT improves the immune microenvironment in aged ovaries,with decreased macrophages and macrophage-derived multinucleated giant cells(MNGCs).These results suggest that FMT from young donors could be a good choice for delaying ovarian aging.

关键词： Ovarian aging Gut microbiota Inflammatory cytokines Ovarian microenvironment

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Effect of Prolactin on Gonadotropin Regulation of Mouse Ovarian Function

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developmental and reproductive biology 1993年第2期2卷 26-32页

作者：胡召元刘以训 State Key Laboratory of reproductive biology Institute of Zoology Academia Sinica Beijing 100080

We have demonstrated that prolactin inhibits gonadotropin-induced ovulation in PMSG-primed *** number of ova in oviducts considerably decreased in the group of hCG plus prolactin (PRL)(19.7 + 4.9) as compared with that of hCG alone (31.7 + 6.7). PRL inhibition of hCG-inducedovulation in mice may be through decreasing the ovarian plasminogen activator (PA) activity on onehand, and inhibiting the preovulatory increase in estrogen (E) secretion on the other.

关键词： Prolactin Mouse Ovulation Plasminogen Activator Estrogen.

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Effect of Forskolin on Progesterone and Plasminogen Activator Secretion by Perfused Rat Corpora Lutea during Pseudopregnancy

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developmental and reproductive biology 1993年第1期2卷 33-40页

作者：祝诚赵芳张丽芳 State Key laboratory of reproductive biology Institute of ZoologAcademia SinicaBeijing 100080

Effects of forskolin on progesterone and plasminogen activator production in pseudopregnant ratcorpora lutea was investigated using isolated in vitro perfused *** andplasminogen activator production were measured on day 1,8 and 18 of *** results indicated:different concentrations of forskolin(100,200,400 and800 μg)administered to ovaries on the 8th day of pseudopregnancy caused elevation of progesteronesecretion in a dose-dependent *** 8 hours of perfusion,PA contents increasedsignificantly in ovaries treated with *** exogenous PA-urokinase(800 U)added to theperfusion solution,progesterone secretion increased significantly as compared to control group andremained on high level throughout the perfusion *** exerting no apparent effects in lowdosage(5 mM),AMCHA,a PA inhibitor,administered in higher dosage(10 and 15 mM)led tomarked reduction in PA activity and progesterone secretion as compared to control *** causes significant elevation of level of progesterone secretion and PA activity inpseudopregnant rat ovary perfused in *** PA seems to regulate progesterone secretion in theperfused rat corpora luteum.

关键词： Forskolin Progesterone Plasminogen activator Corpus luteum

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Cellular mechanisms regulating sperm-zona pellucida interaction

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Asian Journal of Andrology 2011年第1期13卷 88-96页

作者： Andrew T Reid Kate Redgrove R John Aitken Brett Nixon reproductive Science group Discipline of Biological Sciences School of Environmental and Life Sciences University of Newcastle Callaghan NSW 2308 Australia ARC Centre of Excellence in Biotechnology and Development Discipline of Biological Sciences School of Environmental and Life Sciences University of Newcastle CaUaghan NSW 2308 Australia

For mammalian spermatozoa to exhibit the ability to bind the zona pellucida （ZP） they must undergo three distinct phases of maturation, namely, spermatogenesis （testis）, epididymal maturation （epididymis） and capacitation （female reproductive tract）. An impressive array of spermatozoa surface remodeling events accompany these phases of maturation and appear critical for recognition and adhesion of the outer vestments of the oocyte, a structure known as the ZP. It is becoming increasingly apparent that species-specific zona adhesion is not mediated by a single receptor. Instead, compelling evidence now points toward models implicating a multiplicity of receptor-ligand interactions. This notion is in keeping with emerging research that has shown that there is a dynamic aggregation of proteins believed to be important in sperm-ZP recognition to the regions of sperm that mediate this binding event. Such remodeling may in turn facilitate the assembly of a multimeric zona recognition complex （MZRC）. Though formation of MZRCs raises questions regarding the nature of the block to polyspermy, formation and assembly of such a structure would no doubt explain the strenuous maturation process that sperm endure on their sojourn to functional maturity.

关键词： capacitation fertilization spermatozoa sperm-zona pellucida interaction

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