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   Table of Contents - Current issue
Coverpage
January-March 2020
Volume 4 | Issue 1
Page Nos. 1-62

Online since Thursday, April 2, 2020

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EXPERT REVIEWS  

Current status and recent advances in preimplantation genetic testing for structural rearrangements p. 1
Shuo Zhang, Cai-Xia Lei, Xiao-Xi Sun, Cong-Jian Xu
DOI:10.4103/2096-2924.281855  
Preimplantation genetic testing (PGT) is an early form of prenatal genetic diagnosis, which can identify the abnormal embryos cultured in vitro, allow only transfer of genetically normal embryos, and improve the pregnancy rate. In recent years, the rapid development of microarrays and next-generation sequencing (NGS) technologies has remarkably accelerated the clinical application of PGT. In particular, a variety of detection methods have emerged and achieved significant progress in PGT for structural rearrangements (PGT-SR). The detection-related abilities of these methods range from the detection of limited chromosome aneuploidy to comprehensive chromosome screening of the whole genome to differentiation of embryos with normal or balanced translocation/inversion karyotypes. In this study, we reviewed PGT-SR-related detection techniques to provide a better reference for clinical application and research. We have also discussed the potential development of novel techniques in the future.
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Single-cell RNA expression profiling of ACE2 and AXL in the human maternal–Fetal interface Highly accessed article p. 7
Qing-Liang Zheng, Tao Duan, Li-Ping Jin
DOI:10.4103/2096-2924.278679  
2019 novel coronavirus disease has resulted in thousands of critically ill patients in China, which is a serious threat to people's life and health. Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) was reported to share the same receptor, angiotensin-converting enzyme 2 (ACE2), with SARS-CoV. Here, based on the public single-cell RNA-sequencing database, we analyzed the mRNA expression profile of putative receptor ACE2 and AXL receptor tyrosine kinase (AXL) in the early maternal–fetal interface. The result indicates that the ACE2 has very low expression in the different cell types of early maternal–fetal interface, except slightly high in decidual perivascular cells cluster 1 (PV1). Interestingly, we found that the Zika virus (ZIKV) receptor AXL expression is concentrated in perivascular cells and stromal cells, indicating that there are relatively more AXL-expressing cells in the early maternal–fetal interface. This study provides a possible infection route and mechanism for the SARS-CoV-2- or ZIKV-infected mother-to-fetus transmission disease, which could be informative for future therapeutic strategy development.
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CLINICAL GUIDELINE Top

Diagnostic guidelines for infertility p. 11
Zi-Jiang Chen, Jia-Yin Liu, He-Feng Huang, Jie Qiao, Can-Quan Zhou, Guo-Ning Huang, Ying-Pu Sun, Dong-Zi Yang, Xiao-Yan Liang, Qi Yu, Yun Sun, Zheng Li, Li-Qing Fan, Cong-Jian Xu, Yuan-Hua Huang, Xue-Hong Zhang, Jing Yang, Shao-Ming Lu, Lin-Lin Cui, Jun-Hao Yan, Jin-Fang Lin
DOI:10.4103/2096-2924.281853  
Infertility seriously endangers the reproductive health of women at childbearing age. It is defined as the failure to achieve successful pregnancy after 1 year or more of regular unprotected intercourse. Broadly defined, infertility includes two aspects – failure to conceive or have a live birth. This guideline only addressed content relevant to the former. It was proposed by the gynecological endocrine group of the Chinese Society of Obstetrics and Gynecology, Chinese Medical Association, based on relevant guidelines of the World Health Organization, the American Society for Reproductive Medicine, the National Institute for Health and Clinical Excellence, as well as the clinical practice in China. The guideline was reviewed by experts and doctors from medical institutions at all levels, which is applicable to the diagnosis of infertility by physicians in obstetrics, gynecology, and andrology at various medical institutions nationwide.
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ORIGINAL ARTICLES Top

Vitrification of In vitro-matured Oocytes: Effects of meiotic spindle morphology on clinical outcome p. 18
Rui-Huan Gu, Zhi-Chao Li, Jing-Wen Lang, Hua Chen, Yun Feng, Song Guo, Jing Fu, Xiao-Xi Sun, Yi-Juan Sun
DOI:10.4103/2096-2924.281854  
Objective: The meiotic spindle controls chromosome movement and mediates various functions essential for fertilization and early postfertilization events. This study aimed to examine whether vitrification causes meiotic damage in vitro- matured metaphase II (MII) human oocytes, and whether the meiotic spindle morphology influences the subsequent developmental outcomes. Methods: The spindle characteristics of MII human oocytesin vitro matured were studied before and after vitrification using PolScope imaging and immunofluorescence staining. The developmental competence of oocytes was also examined. Results: A total of 419 human MII oocytes were obtained from 593 intracytoplasmic sperm injection cycles at our hospital. Of these oocytes, 54 were used for immunofluorescence staining, whereas the other oocytes were examined by PolScope imaging and classified into three groups according to the meiotic spindle morphology: (A) normal morphology, (B) weak refraction and short meiotic spindle, and (C) no detectable meiotic spindle. The three groups demonstrated statistically significant differences in terms of survival after vitrification. However, differences were not found in terms of oocyte chromosome structure and meiotic spindle morphology on immunofluorescence staining performed before and after vitrification. Oocyte survival, fertilization, and early embryonic development rates were significantly higher in Group A than in Groups B and C with or without vitrification. While vitrification had no effect on these metrics in Group A, Groups B and C demonstrated significantly lower fertilization and cleavage rates after vitrification/warming. Conclusions: Screening for normal meiotic spindle morphology and chromosome configuration before vitrification may increase the yield of healthy viable oocytes for various assisted reproductive technologies.
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Effects of vitrification on the imprinted gene Snrpn in neonatal placental tissue p. 25
Jian-Feng Yao, Yan-Fang Huang, Rong-Fu Huang, Su-Xia Lin, Cai-Qiong Guo, Cheng-Zhou Hua, Pei-Ya Wu, Ji-Feng Hu, You-Zhu Li
DOI:10.4103/2096-2924.281851  
Objective: To investigate the effects of vitrification on the expression of the imprinted gene Snrpn in neonatal placental tissue.Methods: Neonatal placental tissue was collected from women with natural pregnancy (control group) and from women in assisted reproductive technology (ART) pregnancy group, following fresh and vitrified embryo transfer (fresh group and vitrified group, respectively). Snrpn mRNA expression and SNRPN protein levels in placental tissue from these three groups were assessed by real-time reverse transcription polymerase chain reaction and Western blot, respectively. DNA methylation in the Snrpn promoter region was analyzed by bisulfite-pyrosequencing.Results: The expression of Snrpn mRNA and SNRPN protein was found to be higher in placental tissue from the fresh and vitrified ART groups, compared to the control group. There was no significant difference in SNRPN gene or protein expression between the fresh and vitrified groups. DNA methylation at the Snrpn promoter region was not significantly different between these three groups.Conclusions: Human ART may alter the transcriptional expression and protein levels of the imprinted gene Snrpn. However, compared to other ART methods, vitrification may not aggravate or reduce this effect. Moreover, the altered expression of Snrpn is likely not directly related to DNA methylation of the Snrpn promoter region.
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In vitro fertilization with single-Nucleotide polymorphism microarray-based preimplantation genetic testing for aneuploidy significantly improves clinical outcomes in infertile women with recurrent pregnancy loss: A randomized controlled trial p. 32
Yi-Lun Sui, Cai-Xia Lei, Jiang-Feng Ye, Jing Fu, Shuo Zhang, Lu Li, Xian-Dong Peng, Yue-Ping Zhang, Guo-Wu Chen, Xiao-Xi Sun
DOI:10.4103/2096-2924.281852  
Objective: To evaluate the effect of preimplantation genetic testing for aneuploidy (PGT-A) in infertile patients with recurrent pregnancy loss (RPL). Methods: A prospective randomized clinical trial was performed in a university-affiliated fertility center in Shanghai, China. Patients in the PGT-A group underwent blastocyst biopsy followed by single-nucleotide polymorphism microarray-based PGT-A and single euploid blastocyst transfer, whereas patients in the control group underwent routinein vitro fertilization/ICSI procedures and frozen embryo transfer of 1–2 embryos selected according to morphological standards. Results: Two hundred and seven infertile patients with RPL were included in this study and randomly assigned to either the control or the PGT-A group. Baseline variables and cycle characteristics were comparable between the two groups. The results showed that PGT-A significantly improved the ongoing pregnancy rate (55.34% vs. 29.81%) as well as the live birth rate (48.54% vs. 27.88%) and significantly reduced the miscarriage rate (0.00% vs. 14.42%) on a per-patient analysis. A significant increase in cumulative ongoing pregnancy rates over time was observed in the PGT-A group. Subgroup analysis showed that the significant benefit diminished for patients who attempted ≥2 PGT-A cycles. Conclusions: PGT-A significantly improved the ongoing pregnancy and live birth rate, while reduced miscarriage rate in infertile RPL patients. However, the significance diminished in patients attempting ≥2 cycles; thus, further studies are warranted to explore the most cost-effective number of attempts in these patients to avoid overuse.
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Implementation of international society guidelines on chorionicity determination in twins: A multi-Center cohort study in mainland China p. 42
Su-Wen Wu, Qiong-Jie Zhou, Xi-Rong Xiao, Yu Xiong, Huan Liang, Jie Shen, Jon Barrett, Hong Wang, Xiao-Tian Li
DOI:10.4103/2096-2924.281857  
Objective: Ultrasound determination of chorionicity is poor in early pregnancy in China. In an effort to increase the accuracy rate of prompt chorionicity determination, clinical training was provided to primary care physicians. This study assesses the effects of implementing clinical guidelines on chorionicity determination. Methods: A multi-centered cohort study was conducted between January 2014 and June 2017 in 12 hospitals without fetal medicine centers. In 2014, the obstetricians and ultrasound physicians were trained in clinical practice and ultrasound examination relating to chorionicity determination. Linear and binary regression analyses were conducted to identify the effects of introducing the new protocols, including the diagnosis rate of chorionicty and perinatal outcomes, taking the data from 2014 as a baseline. Pregnancy outcomes were additionally adjusted for maternal age. Results: During the period of this study, 3,599 twin pregnancies from 12 centers were enrolled, and a total of 2,998 twin pregnancies were extracted. The rate of overall chorionicity determination, including antenatal and postpartum diagnosis, increased successively from 49.5% in 2014 to 93.5% in 2017 (P < 0.0001). The rate of ultrasonic chorionicity diagnosis before 14 weeks increased from 25.2% in 2014 to 65.0% in 2017 (P < 0.0001). These changes were associated with decreasing incidence of preterm birth, a lower risk of stillbirth, whether for one (P = 0.0456 in 2016) or two fetuses (P = 0.0470 in 2016; P = 0.0042 in 2017) and a decreased rate of admission to neonatal intensive care unit (43.0% in 2014, 37.4% in 2017; P = 0.0032). Conclusions: The implementation of a clinical practice guideline improved both overall and early chorionicity determinations. Regular training workshops of antenatal care are recommended to further promote capability in clinical diagnosis and treatment.
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REVIEW ARTICLE Top

Innate lymphoid cells in normal pregnancy and pregnancy-Related diseases p. 53
Cheng-Jie Wang, Yi Yu, Ming-Qing Li, Wei-Rong Gu
DOI:10.4103/2096-2924.281858  
Innate lymphoid cells (ILCs) are a group of lymphocytes without diversified antigen receptors encoded by gene rearrangement on T and B cells. ILCs, which are tissue-resident innate immune cells, expressed particularly in the mucosa or the barrier surface, contribute to the formation of lymphoid organs, the maintenance of tissue homeostasis, and the regulation of antimicrobial defenses. It has been recently reported that ILCs were enriched at the maternal–fetal interface. During a successful pregnancy, the maternal immune system must tolerate a fetus as an allograft. With the new defined of ILCs, a number of studies have shown that three types of ILCs are involved in embryonic development and pregnancy maintenance as well as the occurrence and development of pregnancy-related complications. This article reviews the types and roles of ILCs in normal pregnancy and pregnancy-related diseases.
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CASE REPORT Top

Successful pregnancy following transfer of embryo with dissolved zona pellucida p. 60
Hong-Xing Li
DOI:10.4103/2096-2924.281856  
This study aimed to describe a case of a patient with the spontaneous dissolution of zona pellucida (ZP) during cleavage embryo development after intracytoplasmic sperm injection (ICSI). This patient experienced early abortion due to fetal chromosome 8 abnormality and received ICSI at 31 years of age. Time-lapse monitoring of embryo cleavage and blastocyst culture was performed, and we found that the ZP of embryos 1, 2, 4, and 5 dissolved gradually during the cleavage stage of embryo development. Five blastocysts that developed from 2 pronuclear zygotes were analyzed using next-generation sequencing, and one frozen-thawed blastocyst was transferred, resulting in intrauterine pregnancy. The results of amniotic fluid puncture showed normal fetal chromosomes. To our knowledge, this is the first report of ZP dissolution during the cleavage-stage embryo; however, the exact mechanism remains unknown. The morphological changes inferred that the granular matter in the periocular space was related to the dissolution of ZP through time-lapse observation.
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