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   Table of Contents - Current issue
Coverpage
July-September 2020
Volume 4 | Issue 3
Page Nos. 129-193

Online since Tuesday, September 29, 2020

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ORIGINAL ARTICLES  

Mageb4, a testis-specific gene, is dispensable for mouse spermatogenesis p. 129
Sheng Gao, Da-Min Yun, Li-Wei Zhou, Yun-Hao Wu, Deng-Feng Lin, Xiao-Long Wu, Fei Sun
DOI:10.4103/2096-2924.296546  
Objective: It has recently been shown that the melanoma antigen gene (MAGE) family is expressed in various tumor cell lines but silent in normal tissues, except germ cell lines. Mageb4, a member of the MAGE family, is highly expressed in the testis and homologous in humans and mice. Whole-exome sequencing studies have identified Mageb4 as a possible X-linked cause of inherited male infertility. However, the function of Mageb4 protein remains largely unknown. Methods: Using clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) 9 technology, we generated a Mageb4 knockout mouse model (Mageb4−/Y) to explore the role of this gene in spermatogenesis. Results: First, immunostaining of testicular cells showed that Mageb4 is localized in the cytoplasm of spermatogonia. Second, Mageb4−/Y male mice displayed significant increases in apoptosis. However, Mageb4−/Y male mice showed normal fertility, including normal sperm concentration, sperm motility, and testicular and epididymal histology. Conclusions: These findings suggest that, despite testis-exclusive expression, Mageb4 is dispensable for mouse spermatogenesis. Future research should focus on the role of this gene in apoptosis, aiming to provide clinical guidance regarding male infertility.
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Differentiation of human adipose-derived stem cells into endometrial epithelial cells p. 137
Fang Yang, Wan-Lin Zhang, Shu-Qiang Chen, Hui-Jun Sun, Jie Lu, Xi-Feng Xiao, Xiao-Hong Wang
DOI:10.4103/2096-2924.296547  
Objective: This study aimed to investigate the differentiation of human adipose-derived stem cells (hASCs) into endometrial epithelial cells (EECs) under certain induction conditions and to a further step provide a promising approach for ASCs in clinical practice to the treatment of severe intrauterine adhesion. Methods: Four groups of hASCs were separately cultured as follows: in Group 1, hASCs were cultured in a control medium (5% fetal bovine serum [FBS] + α-minimum Eagle's medium [α-MEM]); in Group 2, hASCs were cultured in an induction medium (5% FBS + α-MEM + [1 × 10−7 mol/L 17β-estradiol] + 10 ng/mL transforming growth factor β1 [TGF-β1] + 10 ng/mL epidermal growth factor [EGF] + 10 ng/mL platelet-derived growth factor BB [PDGF-BB]); in Group 3, hASCs and human endometrium cells (hEMCs) were cocultured in the control medium; and in Group 4, hASCs and hEMCs were cocultured in the induction medium. Results: When cocultured with hEMCs, the morphology of hASCs became similar with EECs, and the addition of factors such as EGF, TGFβ, PDGF-BB, and 17β-estradiol promoted differentiation. This study, for the first time, demonstrated estrogen receptor (ER)α and ERβ expression in hASCs and preliminarily explored changes in ERα, ERβ, β-catenin, and H19 mRNA expression during hASC differentiation. Furthermore, we concluded that H19 mRNA expression was negatively correlated with differentiation, which is seemingly related to the estrogen signaling pathway. Conclusions: hASCs revealed the potential for differentiating to EECs when cocultured with hEMCs.
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Supplementation with L-carnitine rescues sperm epigenetic changes in asthenospermic male semen with altered acetyl-L-carnitine levels p. 146
Xiao-Hui Jiang, Chuan Jiang, Lin Yu, Xiao-Liang Li, Tao Zuo, Pei-Fei Gu, Fu-Ping Li, Wen-Ming Xu
DOI:10.4103/2096-2924.294314  
Objective: To investigate the relationship between the concentration of L-carnitine in semen and sperm parameters and investigate the epigenetic profile in sperm cell after L-carnitine usage. Methods: From February 2017 to February 2018, 46 semen samples from asthenospermic males and 41 semen samples from healthy donors were acquired. Motility parameters were assessed using computer-assisted sperm analysis (CASA, n = 78) and the DNA fragmentation index (DFI) was evaluated through flow cytometry (n = 86), %DFI = % cells outside main population. Other oxidative stress markers, such as reactive oxygen species (ROS) levels (n = 86) and the mitochondria DNA copy numbers, were detected (n = 78). The concentration of L-carnitine and acetyl-L-carnitine was detected (n = 82), and methylation was analyzed (n = 30). After that, we collected 13 fresh semen samples from asthenospermic males and 23 fresh semen samples from healthy donors. These samples were used in a freeze-thaw model that was used to determine whether adding L-carnitine could change sperm progressive motility (n = 23), apoptosis index (n = 9), and methylation analysis (n = 7). In total, we have done 13 asthenospermia samples for Western blot, and except for the poor Western result, we analyzed 6 samples for H3K9ac detection, 7 samples for H3K9m3 and H3K27m3 detection, and immunofluorescence (n = 3). Finally, we had recruited 30 volunteers, and they were given oral administration of L-carnitine for 3 months and then collected semen samples at different time points for methylation analysis. Results: The concentration of acetyl-L-carnitine is negatively correlated with the %DFI value (r2 = 0.1090; P = 0.0026), and the concentration of acetyl-L-carnitine is positively correlated with sperm forward motility (r2 = 0.0543; P = 0.0458) and ROS (r2 = 0.1854;P < 0.0001), and the acetyl-L-carnitine level is negatively correlated with %DFI in asthenospermia (r2 = 0.1701; P = 0.0066), and the level of acetyl-L-carnitine in asthenospermic semen is significantly lower than the normal group (P = 0.0419). In addition, this study indicates that adding L-carnitine significantly improved sperm motility (P = 0.0325) and reduced sperm apoptosis (P = 0.0032). Importantly, Western blotting (P = 0.0429) and immunofluorescence staining results showed that the addition of L-carnitine reduced H3K9Me3 methylation level in sperm, respectively. Furthermore, semen samples from asthenospermic patients had reduced methylation levels in a specific region (16th P = 0.0003; 17th P = 0.0016) of the brain-derived neurotrophic factor (BDNF) promoter. The 16th methylation decreased with age (r2 = 0.1564; P = 0.0306), and the 17th methylation was decreased after treatment with L-carnitine for 28 days (P = 0.0341). Conclusion: L-carnitine can reduce the %DFI and also affect the methylation of the histone modification marker in sperm as a possible epigenetic regulator.
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A modified ultra-long downregulation protocol improves pregnancy outcomes in high body mass index patients undergoing In vitro fertilization/intracytoplasmic sperm injection treatment p. 156
Hui-Jun Chen, Yuan Li, Xiao-Feng Li, Ge Lin, Guang-Xiu Lu, Fei Gong
DOI:10.4103/2096-2924.294315  
Objective: Overweight and obesity are increasingly epidemic and negatively related to reproductive outcome. The aim of this study was to investigate the advantages of a modified ultra-long downregulation protocol on pregnancy outcomes of patients with high body mass index (BMI) undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI), compared to the long protocol (LP). Methods: We retrospectively analyzed the clinical data of 3,920 infertile patients at the Reproductive and Genetic Hospital of Citic-Xiangya from January 2012 to December 2017 by propensity score matching (PSM). Patients were divided into two groups: modified ultra-LP (MULP) (n = 1,960) and LP (n = 1,960). Results: In the MULP group, live birth rate (52.65% vs. 46.79%,P < 0.001, odds ratio [OR]: 1.784, 95% confidence interval [CI]: 1.563–2.036), clinical pregnancy rate (62.50% vs. 57.91%, P = 0.003, OR: 1.211, 95% CI: 1.066–1.377), and implantation rate (53.24% vs. 49.65%, P = 0.004, OR: 1.155, 95% CI: 1.048–1.272) were statistically significantly higher than those of the LP group. Moreover, the cycle cancellation rates (12.70% vs. 15.15%, P = 0.027, OR: 0.815, 95% CI: 0.68–0.977), abortion rates (12% vs. 14.8%, P = 0.046, OR: 0.785, 95% CI: 0.619–0.996), and ectopic pregnancy rates (1.06% vs. 2.11%, P = 0.04, OR: 0.497, 95% CI: 0.252–0.98) were lower than those in the LP group. Conclusion: The modified ultra-long downregulation protocol improved the pregnancy outcomes in patients with high BMI undergoing IVF/ICSI treatment, providing a potential option for physicians when deciding an optimized ovary stimulation protocol for high BMI patients.
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Clinical performance of cell-free fetal DNA testing for fetal aneuploidies and subchromosomal deletions/duplications in a cohort of 19,531 pregnancies p. 163
Yi-Sheng Chen, Yong-Qin Wu, Ying Zhang, Chun-Mei Ying
Objective: We aim to assess the clinical performance of cell-free fetal DNA (cffDNA) testing for detecting common fetal aneuploidies as well as subchromosomal deletions/duplications and explore the pregnancy decisions in screen-positive cases. Methods: A cohort of 19,531 pregnant women was offered cffDNA testing for detection of trisomies 21, 18, and 13 (T21, T18, and T13); sex chromosome aneuploidies (SCAs); and subchromosomal deletions/duplications. Screen-positive cases were confirmed by karyotyping and single-nucleotide polymorphism array analysis. Results: A total of 47 cases failed the test. The overall screen-positive rate of chromosomal abnormalities was 1.07% (208/19,484), including 57 cases with T21, 18 cases with T18, 7 cases with T13, 106 cases with SCAs, and 20 cases of subchromosomal deletions/duplications. Positive predictive values were 91.30% (42/46), 38.46% (5/13), 33.33% (2/6), 41.33% (31/75), and 27.78% (5/18), respectively. There was no significant difference in the screening of fetal chromosomal aneuploidies in the high-risk group compared with the low-risk group (P > 0.05). All of the pregnant women who had confirmed fetal T21, T18, or T13 terminated their pregnancies, except for a case of T13 mosaic, whereas 45.16% (14/31) of women with fetal SCAs continued their pregnancies. Furthermore, 17 pregnant women with positive screens for T21, T18, or T13 without a subsequent diagnosis chose to terminate their pregnancy, whereas 29 of 31 women with SCAs chose to continue their pregnancies. Conclusions: CffDNA testing exhibited good screening accuracy for T21, T18, and T13 and also contributed to detecting fetal SCAs and subchromosomal deletions/duplications. Pregnant women with fetal 47, XXX or 47, XYY were more willing to terminate their pregnancy than those with fetal 45, X or 47, XXY.
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Effects of endometrial stimulation timings and techniques on pregnancy outcomes in patients without prior embryo transfer: A systematic review and meta-analysis p. 169
Yu-Rong Cao, Hao Shi, Jun Zhai
DOI:10.4103/2096-2924.296549  
Objective: To investigate the effects of endometrial stimulation timings and techniques on pregnancy outcomes in patients without prior embryo transfer (ET). Methods: We included a total of 10 studies related to the impact of endometrial stimulation on the pregnancy outcome of infertile patients with the first ET from 2010 to 2019. These studies were found by searching databases including China Science and Technology Journal Database (VIP), Chinese Biological Med (CBM), Chinese Medical Current Content (CMCC), China National Knowledge Internet (CNKI), WanFang Med Online, Cochrane Library, Web of Science, PubMed, Medline, ScienceDirect, and EMBASE. A total of 1,983 cycles were included, of which 725 were cycles with endometrial stimulation. Clinical outcomes included clinical pregnancy, implantation, abortion, multiple pregnancy, and live birth rate. Results: The implantation rate (IR) was higher in the fresh cycle endometrial stimulation group than in the control group (relative risk [RR] = 1.21, 95% confidence interval [CI] = 1.03–1.42; P = 0.02), but there were no significant between-group differences in the live birth rate (LBR) and abortion rate (AR). Subgroup analysis showed that whether follicular or luteal endometrial stimulation was performed before the ET cycle had no effect on the clinical pregnancy outcome, and endometrial stimulation on the day of oocyte retrieval reduced the clinical pregnancy rate (CPR) (RR = 0.37, 95% CI = 0.19–0.75; P = 0.005). Whether the technique involved the use of a curette or catheter, there was no significant between-group difference in CPR. Conclusions: Fresh cycle endometrial stimulation can improve the embryo IR in patients without prior ET, but it cannot increase CPR, LBR, or AR. Subgroup analysis showed that different endometrial stimulation timings and techniques did not significantly improve CPR and that endometrial stimulation on the day of oocyte retrieval reduced CPR.
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Comparative neonatal outcomes of vitrified versus fresh embryo transfers: A systematic review and meta-analysis p. 177
Cai-Hong Li, Dong-Kai Cheng, Hong-Jun Yu, Chun-Yi Li, Hai-Qin Ren, Ning Weng, Bao-Shan Li, Na Yang, Peng Xu
DOI:10.4103/2096-2924.288961  
Objective: Vitrification prevents ice formation within the high concentration of cryoprotectant agents and allows the cells and extracellular milieu to solidify into a glass-like state. This study aimed to provide objective information on the available data regarding singleton births comparing embryo vitrification with fresh embryo transfers. Methods: PubMed and Cochrane databases were searched for studies conducted up to 2017. Low birth weight (LBW), very low birth weight (VLBW), preterm birth (PTB), very PTB, small for gestational age (SGA), large for gestational age, perinatal mortality, cesarean section, and preeclampsia were measured. Results: The search returned 220 articles, and 13 of these met the inclusion criteria. A significantly lower risk of SGA birth was observed in the vitrified embryo transfer group than in the fresh transfer group. Conversely, a higher risk of very PTB, VLBW, large size for gestational age, and perinatal mortality was observed more frequently in the vitrified embryo transfer group. Conclusions: Vitrified embryo transfer was associated with the improved outcome of neonates regarding the SGA rate compared with fresh embryo transfer, but included a higher risk of very PTB, VLBW, large size for gestational age, and perinatal mortality. Caution should be exercised in interpreting these findings given the low level of evidence of the studies.
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REVIEW ARTICLE Top

The application of aspirin in pregnancy-related complications p. 184
Lu-Yu Ruan, Zhen-Zhen Lai, Hui-Li Yang, Shao-Liang Yang, Si-Yao Ha, Jia-Wei Shi, Hui-Hui Shen, Ming-Qing Li
DOI:10.4103/2096-2924.296548  
Aspirin, one of the most widely applied medicines, not only possesses the effects on reducing fever, anti-vascular hyperplasia, and anti-inflammation, but also has the capacity of preventing platelet aggregation. So far, it is acceptable to adopt aspirin, especially low-dose aspirin (LDA), to prevent pregnancy-related complications, such as pregnancy complicated by antiphospholipid syndrome, systemic lupus erythematosus, or preeclampsia; unexplained recurrent spontaneous abortion; fetal growth restriction; and preterm birth. In this article, we reviewed the possible mechanism of action and applications of aspirin in these pregnancy-related complications.
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CASE REPORT Top

Postpartum spontaneous rupture of an ovarian artery aneurysm: A case report and review p. 191
Hui-Fen Yin, Hai-Yan Liu, Wen-Tao Jin, Rong Hu
DOI:10.4103/2096-2924.296545  
This study reported a case of spontaneous rupture of an ovarian artery aneurysm 4 days following cesarean section. The patient mainly presented with a consistent low-grade fever and did not have symptoms, such as severe abdominal or flank pain, as is common in other similar cases. Enhanced magnetic resonance imaging revealed a massive retroperitoneal hematoma; angiography confirmed that there was a rupture in an ovarian artery aneurysm. A selective embolization was performed successfully.
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