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   Table of Contents - Current issue
Coverpage
April-June 2020
Volume 4 | Issue 2
Page Nos. 63-127

Online since Friday, June 26, 2020

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ORIGINAL ARTICLES  

Mifepristone (RU486) inducing abortion in a mouse model by regulating innate and adaptive immune responses p. 63
Meng-Die Li, Yi-Fei Sang, Yun-Hui Tang, Ling Xu, Chun-Fang Xu, Da-Jin Li, Yan-Hong Li, Jian-Ping Zhang, Mei-Rong Du
DOI:10.4103/2096-2924.288021  
Objective: Mifepristone (RU486), one of the most common medications for artificial abortion, attenuates the immunoregulatory effects of progesterone. However, the specific immune regulatory mechanism of RU486 in abortion remains unknown. We intended to investigate the immunomodulatory effects of RU486 on abortion. Methods: Sixty female mice were divided into the control group (0 mg RU486) and RU486 group (2 mg/kg RU486). The uterus, peripheral blood, and spleen were obtained for isolation of specific cell types. The population and phenotype of immune cells in the decidua, peripheral blood, and spleen were analyzed using flow cytometry. Statistical differences between groups were determined using two-tailed t- test. For all statistical tests, P < 0.05 was considered statistically significant. Results: RU486 effectively induced abortion in pregnant mice, with a significantly higher number of decidual macrophages (dMφ) (control group = 25.55% ± 2.467%, RU486 group = 19.41% ± 1.423%; P < 0.05), especially the major histocompatibility complex IIhigh subset. No difference in Mφ number was observed in the spleen or peripheral blood. Moreover, the dMφ from mice with RU486-induced abortion displayed a remarkable activated phenotype, with increased expressions of inducible nitric oxide synthase, tumor necrosis factor-α, and interleukin (IL)-12 but decreased expressions of arginase-1 and IL-10. We also found elevated levels of decidual CD4+ T-cells in the RU486 group that exhibited a higher level of the proinflammatory cytokine interferon-γ and a lower level of the anti-inflammatory cytokines, IL-4 and IL-10. Conclusions: We report a new mechanism of RU486-induced abortion via the regulation of innate cell Mφ activation and the adaptive response of CD4+ T-cells present in the decidua but not the periphery.
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Homozygous mutation in the MTHFS gene may contribute to the development of cerebral folate deficiency syndrome p. 72
Dharaniya Sakthivel, Yunping Lei, Xuanye Cao, Richard H Finnell
DOI:10.4103/2096-2924.288022  
Objective: The purpose of this study was to examine the role of rare variants in the one-carbon metabolic pathway in the etiology of the cerebral folate deficiency (CFD) syndrome. The CFD syndrome is a neurometabolic syndrome identified by low concentrations of 5-methyltetrahydrofolate (5-MTHF) in the cerebrospinal fluid (CSF) in spite of near-normal peripheral folate levels resulting in neurodevelopmental disorders. Methods: The localized folate metabolism impairments in CFD are thought to be either the result of mutations in genes responsible for folate transport or folate turnover through degradation. Genes that have been previously implicated in the etiology of CFD include folate receptor alpha-1 (FOLR1), dihydrofolate reductase, proton-coupled folate transporter, and capicua. We performed whole-exome sequencing (WES) analysis of a CFD patient that revealed 99 novel missense mutations, of which 21 were classified as damaging mutations through the Poly-Phen2 prediction algorithm. In vitro functional studies were conducted by transient transfection of wild-type and mutant MTHFS into HEK293T cells to determine the impact of the variants on enzyme activity. Results: Of the damaging variants identified in the WES studies, we focused on the gene coding for the enzyme 5,10-methenyl-tetrahydrofolate synthetase (MTHFS). This enzyme catalyzes the production of methenyl THF which is subsequently converted to 5-MTHF. The CFD patient described within was found to carry a homozygous mutation, c.101G>T (p.R34L, rs200058464) in MTHFS, while the parents of the proband are heterozygotes for the MTHFS gene, and the healthy sibling is not a carrier. Conclusion: The mutant allele displayed a 50% reduction in luciferase activity (P < 0.05), suggesting that homozygous loss of the MTHFS gene may play a significant role in the development of CFD.
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Pan-specific antibodies as novel tools to detect valyllysine p. 78
Ya-Ling Wang, Min-Yan Liu, Yu-Hua Li, Yi-Ting Yang, Wei-Wei Wang, He-Guo Yu, Zhi-Yu Shao, Hua Diao
DOI:10.4103/2096-2924.288018  
Objective: Amino acyl modification of lysine residues is an essential mechanism of nutrient sensing that regulates various biological functions including reproduction. At present, the lack of pan-specific antibodies for a recently identified lysine valylation hinders the characterization and detection of this modification. The objective of this study is to raise pan-specific antibodies that may facilitate the identification of novel expression patterns of lysine valylation. Methods: Chicken ovalbumin was valylated as an immunogen to raise polyclonal antibodies (PcAbs) in rabbits. The population of the pan-specific antibodies recognizing valylated lysine was purified using the chemically synthesized valylated peptides consisting of random amino acids. The specificity of the antibodies was evaluated using ELISA, dot blots, Western blots, and immunohistochemistry (IHC) staining in human epididymis as well. Results: A preliminary and simple strategy to make an anti-valylated lysine PcAb was developed. The recognition of the antibodies to valyllysine was evaluated as pan specific. This was useful for the detection of the newly identified valyl modification using ELISA, dot blots, and Western blots. The antibodies were also successfully utilized in IHC assays, which revealed novel valyllysine modification patterns in epididymis tissues of human. Conclusions: A new antibody tool was provided for the study of lysine valylation. The novel expression patterns of valyllysine in the epididymis suggest that this modification may be involved in sperm maturation.
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Identification of a novel compound heterozygous mutation in OTOG in a chinese family with severe hearing impairment p. 84
Wen-Ya Yan, Fan Xu, Bing Li
DOI:10.4103/2096-2924.288023  
Objectives: Hearing loss is a worldwide disease. In 50% of the patients, hearing loss is caused by genetic problems associated with GJB2, MTRNR1, SLC26A4, and other genes. Considering the recent development and cost reduction of whole-exome sequencing, it is possible to filter out the normal genes and find which among the more novel genes contributed to the loss of hearing. Methods: After prescreening all individuals for GJB2, MTRNR1 and SLC26A4 mutations, whole-exome sequencing was performed in the proband, and the pathogenic variant was confirmed via Sanger sequencing. Results: The compound-heterozygous variant namely c.8076G>C:p.E2692D and c.6362T>C:p.V2121A in OTOG was identified as a candidate gene of a consanguineous Kazakh family. Conclusion: This is the first reported case of severe deafness caused by an OTOG compound-heterozygous variant in the world and the first case of deafness caused by an OTOG variant in China. This discovery identified the important contribution of OTOG toward deafness and expanded the spectrum of variants responsible for human hearing loss.
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Comparison of the efficacy and safety of different surgical strategies for patients with type II cesarean scar pregnancy p. 89
Qi Cheng, Qi Tian, Kai-Kai Chang, Xiao-Fang Yi
DOI:10.4103/2096-2924.288024  
Objective: To compare the efficacy and safety of four surgical strategies currently used for the management of deep implantation cesarean scar pregnancy (CSP-II). Methods: This was a retrospective clinical cohort study, and, in total, 131 women diagnosed with CSP-II and primarily treated in our hospital were recruited. Women treated using laparoscopy assisted by operative hysteroscopy (LAOH; Group A, n = 25), uterine artery embolization (UAE) followed by LAOH (Group B, n = 21), ultrasound-guided dilatation and curettage (D&C; Group C, n = 24), and UAE followed by D&C (Group D, n = 61) were evaluated. Univariate and multiple logistic analyses were performed to identify the risk factors. Results: No statistically significant difference was found in patient age, gestational age, size of lesion, and pretreatment serum β-human chorionic gonadotropins (β-hCG) level. Operation time was longer (P < 0.001) and the success rate was higher (P = 0.01) in both Group A and Group B than in Group C and Group D. When the cohort was further analyzed regarding patients with myometrial thickness ≤3 mm (n = 75, defined as CSP-IIb), a lower rate of perioperative complications (P = 0.036) and a higher success rate (P < 0.001) remained in Group A (n = 15) and Group B (n = 15) but not in Group C (n = 11) or Group D (n = 34). In multiple logistic regression analysis, the risk factors related to lower treatment efficacy for patients with CSP-II were thinner myometrial thickness of cesarean scar (CS) (≤3 mm) (odds ratio [OR] = 5.470, P = 0.062), number of cesarean sections (a2) (OR = 8.877, P = 0.013), mass protruding into the bladder or abdominal cavity (OR = 25.507, P < 0.001), and direct D&C modality (OR = 38.247, P = 0.010). Conclusions: Compared with D&C ± UAE, LAOH ± UAE showed a higher success rate for patients with CSP-II, especially when the zygote was more deeply implanted with a myometrial thickness of CS ≤ 3 mm. CSP-II treatment should be individualized on the basis of many risk factors.
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Effect of progestin-primed ovarian stimulation protocol in infertile women with basal follicle-stimulating hormone levels ≥15 IU/L: A retrospective analysis p. 97
Lu Fang, Xiu-Juan Qi, Hong Zhu
DOI:10.4103/2096-2924.285780  
Objective: To evaluate the efficacy of progestin-primed ovarian stimulation (PPOS) protocol in infertile women with high basal follicle-stimulating hormone (FSH) levels ≥15 IU/L. Methods: Patients with high basal FSH levels ≥15 IU/L with autologous oocytes from September 2016 to March 2019 were reviewed. Either medroxyprogesterone acetate 4 mg/d or clomiphene citrate (CC) 50 mg/d was administered daily from day 3 to the trigger day. When serum FSH levels decreased to ≤15.0 IU/L, a low dose of human menopausal gonadotropin (hMG) 75/150 IU/d was administered to promote late follicular development. Results: Two hundred and twenty women were retrospectively analyzed in this study. Among them, 139 patients were administered with PPOS protocol as the study group, and 81 patients were administered with CC protocol as the control group. The numbers of received oocytes and viable embryos were higher in the study group than those in the control group (1.5 ± 1.2 vs. 1.2 ± 0.8 and 0.8 ± 0.8 vs. 0.5 ± 0.6, respectively, P < 0.05). However, hMG duration and dosage were significantly higher in the study group than those in the control group (4.2 ± 2.7 d vs. 1.1 ± 2.3 d and 609.1 ± 424.5 IU vs. 140.7 ± 231.3 IU, respectively, P < 0.01). Incidence of luteinizing hormone surge and cycle cancellation rate were lower in the study group than those in the control group with statistical difference (2.88% vs. 16.05% and 36.50% vs. 50.63%, respectively, P < 0.05). Conclusions: PPOS protocol can effectively downregulate the endogenous FSH levels. Compared with CC protocol, treatment with PPOS protocol in patients with high basal FSH levels ≥15 IU/L could receive more oocytes and more viable embryos.
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Effect of stimulating the BL67 point on fetal correction from breech to cephalic presentation and natural delivery after the 36 weeks of pregnancy: A randomized clinical trial p. 103
Khatereh Sourani, Abolfazl Mohammadbeigi, Nasim Khademi, Azadeh Asgarian, Imaneh Khaki, Zohreh Ahmadi
DOI:10.4103/2096-2924.288019  
Objective: Various techniques are proposed for changing fetal presentation. We aimed to assess the effect of BL67 point stimulation on correcting breech presentation and natural delivery in women at 36–38 gestational weeks. Methods: A parallel single blinded randomized clinical trial was conducted on 72 eligible pregnant women with breech presentation at the 36 weeks of pregnancy. The subjects were divided into two groups – intervention (n = 36) and control groups (n = 36) by block randomization method. The intervention group stimulated the BL67 point by self-administration for 20 min once a day for 2 weeks. Finally, the appearance of cephalic presentation and rate of vaginal delivery was compared between the groups (n 1 = n 2 = 32) using the Chi-square test and multivariate logistic regression. Results: The correction of breech to cephalic presentation occurred in 53.1% of patients in the intervention group. The adjusted relative risk (RR) for fetal correction from breech to cephalic was 1.80 (RR = 1.80, 95% confidence interval [CI], 1.13–5.17). It was shown that the stimulation of the BL67 point increased the correction of breech to cephalic presentation. In addition, the rate of vaginal delivery increased by >4-fold (RR = 4.16, 95% CI, 2.54–6.82) by correction to cephalic presentation. Moreover, 65.6% of mothers in intervention group and 90.6% in the control group underwent cesarean section. Conclusions: The stimulation of the BL67 point is a safe, inexpensive, and effective method that can be self-administered at home for fetal correction from breech to cephalic presentation in women with breech presentations during 36–38 gestational weeks. This promotes uncomplicated natural childbirth.
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REVIEW ARTICLE Top

Functions of lysosomes in mammalian female reproductive system p. 109
Yuehuan Li, Zidao Wang, Christian L Andersen, Xiaoqin Ye
DOI:10.4103/2096-2924.288025  
The lysosome is the most acidic membrane-bound intracellular organelle. Lysosomal acidity is primarily maintained by vacuolar H+-ATPase (V-ATPase) and counter ion channels. There are >60 hydrolytic enzymes in the lysosome for its fundamental digestive role. Lysosomes also play important roles in endocytosis, exocytosis, autophagy, and cell death. Studies that have implicated roles of lysosomes in the female reproductive system are reviewed here. In the ovary, lysosomes are implicated in the preparation of free cholesterol for steroidogenesis and degradation of regulators of steroidogenesis, regulation of follicular atresia, follicle rupture during ovulation, luteal cell survival, and luteal regression. In the oviduct, lysosomes are involved in endocytosis of both serum and oviductal luminal components. In the uterus during the menstrual/estrous cycle, lysosomes are associated with endometrial secretion, apoptosis, and menstruation. In the uterus during early pregnancy, lysosomes are involved in the temporal and directional changes of endocytosis, uterine epithelial acidification upon embryo implantation initiation, and embryo-maternal mutual communications via extracellular vesicles. In the placenta, lysosomes are implicated in nutrient transport and placental separation from the uterus for parturition. In the mammary gland, lysosomes are important for mammary gland development and involution. These findings suggest/demonstrate functions of lysosomes in multiple processes of female reproduction, from ovulation to ovarian steroidogenesis for pregnancy maintenance, and from essential in utero nurturing of developing embryos/fetuses via embryo/fetal-maternal communications, to optional postpartum nurturing of newborns via lactation. Future studies using genetically or modified animal models and pharmacological approaches will provide novel insights into the functions and mechanisms of lysosomes in the mammalian female reproductive system.
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CASE REPORT Top

Acute miliary tuberculosis and pelvic tuberculosis after In vitro fertilization p. 123
Li-Juan Fan, Ting-Ting Ma, Shan Liu, Yun Qi
DOI:10.4103/2096-2924.288020  
Nearly one-fourth of the world's population is infected with Mycobacterium tuberculosis (MTB). Female genital tuberculosis (TB) is a common cause of infertility in both developing and undeveloped countries. Furthermore, assisted reproduction treatments and pregnancy potentially increase the risk of TB infection and reactivation. In this study, we present the case of a 28-year-old infertile female without a history of TB who developed an acute miliary TB and pelvic TB after in vitro fertilization–embryo transfer (IVF-ET). Elevated serum estrogen levels during controlled ovarian hyperstimulation and T-lymphocyte function inhibition during pregnancy are the risk factors for MTB infection and reactivation. In her 7th week of gestation, the patient developed fever and spontaneously aborted. Her chest computed tomography images revealed classical miliary TB. Uterine curettage tissue and vaginal secretion samples as well as Gene X-pert MTB/rifampicin (RIF) and TB-RNA test results were positive for MTB. Histological examination of the uterine curettage tissue confirmed the diagnosis of endometrial TB. Treatment with isoniazid, RIF, pyrazinamide, amikacin, and levofloxacin was selected based on the patient's diagnosis, complications, and test results. Currently, the patient is undergoing anti-TB treatment, and her condition is stable. It is important to rule out the presence of TB in infertile patients before performing IVF-ET to avoid TB dissemination during pregnancy.
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