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   Table of Contents - Current issue
January-March 2019
Volume 3 | Issue 1
Page Nos. 1-62

Online since Thursday, April 11, 2019

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Overview of Oncofertility p. 1
Ding Ma
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Deficiency of antisense lncRNA Gm48853 resulted in embryonic lethality and impaired placental development in mice p. 5
Qi-Yun Qin, Wei-Jia Zeng, Chu-Yue Peng, Yu-Fang Zheng, Hong-Yan Wang
Objective: Long noncoding RNAs (lncRNAs) play important roles in embryonic development and in various diseases. Gm48853 is a predicted natural antisense lncRNA gene of unknown function. The present study aimed to investigate its expression and function. Methods: Quantitative reverse transcription-polymerase chain reaction and sequencing were used to detect the expression of Gm48853 antisense lncRNA. Hematoxylin and eosin staining and pathological analysis were applied to evaluate the effects of Gm48853 mutations on embryonic/placental development. 5-Ethynyl-2'-deoxyuridine (EdU) and terminal deoxynucleotidyl transferase deoxy-UTP-nick-end labeling assays were used to analyze the status of cell proliferation and apoptosis in Gm48853- deficient embryos and placentas. Results: PiggyBac (PB) transposon insertion into the intron of Gm48853 resulted in a significant decrease in Gm48853 expression and embryonic lethality at midgestation. Homozygous Gm48853 mutations led to severe growth retardation of embryos and impaired the morphogenesis of the placental labyrinth. In addition, cell proliferation was dramatically decreased in Gm48853PB/PB embryos and placentas, and apoptosis was radically increased in the mutant embryos but not in the mutant placentas. Conclusion: Antisense lncRNA Gm48853 may regulate embryonic/placental development by enhancing cell proliferation and/or inhibiting apoptosis.
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The messenger RNA and long non-coding RNA expression profiles in ectopic and eutopic endometrium provide novel insights into endometriosis p. 11
Song-Ping Liu, Xin Tian, Hong-Yan Cui, Qiong Zhang, Ke-Qin Hua
Objective: To establish the messenger RNA (mRNA) and long non-coding RNA (lncRNA) expression profiles in ectopic and eutopic endometrium and provide novel insights into endometriosis. Methods: The mRNA and lncRNA expression profiles were tested using high-throughput sequencing technology in ectopic and eutopic endometrium with endometriosis and normal endometrium without endometriosis. The potential targeted lncRNAs were annotated by analyzing the correlation between lncRNA and mRNA expression to better understand the pathogenesis of endometriosis. Results: In ectopic compared with normal endometrium, a total of 2,188 mRNAs and 1,200 lncRNAs were differentially expressed with a fold-change (FC) ≥2.5. In eutopic compared with normal endometrium, a total of 2,324 mRNAs and 695 lncRNAs were differentially expressed with an FC ≥1.5. In ectopic compared with eutopic endometrium, a total of 2,223 mRNAs and 511 lncRNAs were differentially expressed with an FC ≥2. Bioinformatic analysis indicated that the differentially expressed mRNAs were enriched in the biological processes and signaling pathways involved in endometriosis. In addition, we constructed a gene coexpression network based on the dysregulated lncRNAs in both ectopic endometrium and eutopic endometrium, combined with their coexpressed mRNAs to simulate the complex interactions. Conclusions: This study describes the first-to-integrate analysis of the differential expression profiles of mRNAs and lncRNAs, including analyses between ectopic and normal endometrium, eutopic and normal endometrium, and ectopic and eutopic endometrium, which provides new insights to investigate the pathogenesis of endometriosis and explore novel diagnostic biomarkers and therapeutic targets.
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LncRNA4667 is dispensable for spermatogenesis and fertility in mice p. 18
Yu-Bing Dai, Yu Lin, Ning Song, Fei Sun
Objective: Spermatogenesis is a complex process which is of vital importance for sexual reproduction. In many studies of spermatogenesis, the mRNAs, protein-coding genes, as well as small noncoding RNAs (ncRNAs) have been well characterized. However, there remain numerous questions despite previously characterized molecular mechanisms. Long ncRNAs (lncRNAs) are a relatively new addition to our knowledge of ncRNAs. Limited studies have examined the function of lncRNAs in spermatogenesis. Therefore, we selected a testis-specific lncRNA, lncRNA4667, to analyze its potential role in spermatogenesis and male fertility. Methods: In situ hybridization and quantitative reverse transcription polymerase chain reaction analyses were used to confirm testis-specific expression of lncRNA4667. LncRNA4667 knockout mice were generated using CRISPR/Cas9 technology. Histology, sperm counts, sperm motility, body parameters, and fertility were compared between wild-type and knockout mice (n = 8/group). Results: Expression analysis showed that lncRNA4667 was testis specific and localized to round spermatids in seminiferous tubules of adult mouse testes. Mice homozygous for a null mutation of lncRNA4667 displayed normal spermatogenesis and fertility compared with wild-type mice. Conclusions: These data indicate that lncRNA4667 is dispensable for spermatogenesis and fertility in mice, and the localization of lncRNA4667 makes it a useful marker for the identification of round spermatids in mice.
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Clinicopathological evaluation of ovarian juvenile granulosa cell tumor: Is fertility-sparing surgery safe? p. 24
Lu Guo, Xiao-Cheng Liu, Xiu-Ying Chen, Xi-Rong Xiao, Yu-Qing Qu, Bin Li
Objective: To retrospectively investigate the clinicopathological characteristics of ovarian juvenile granulosa cell tumors (JGCTs) and to evaluate the safety of fertility-sparing surgery. Methods: In this study, surgically treated patients with JGCTs diagnosed between January 2004 and October 2018 in our center were identified. Clinicopathological data, survival outcomes, and recurrence rates were examined in these patients. Results: A total of 8 patients were included. All patients were premenarchal girls or young women (age range, 9–32 years). Irregular vaginal bleeding was the most common presenting symptom. Of them, seven patients were classified with Stage I JGCTs, and they underwent fertility-sparing surgery. One patient who had Stage IIIC JGCT and had completed childbearing underwent complete surgery. Seven patients received adjuvant chemotherapy. The median follow-up duration in the total cohort was 64 months (range, 2–117 months). The overall survival rate in the fertility-sparing group was 100%, whereas the patient with Stage IIIC JGCT died 1 month after the treatment. Conclusions: Fertility-sparing surgery might not show a negative impact on oncologic outcomes. Fertility sparing could be considered a modified option for patients with Stage I JGCTs. However, due to the limited number of patients, the conclusion must be interpreted with caution, and larger or multicenter studies are needed before conclusions can be drawn.
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Safety of mifepristone in medical abortion in hyperthyroidism pregnant mice p. 30
Yun-Hui Tang, Xiao-Ying Yao
Objective: To study the safety of mifepristone on thyroid hormone level by using hyperthyroidism pregnant model in mouse to simulate the process of medical abortion and observe the changes of thyroid hormone during abortion. Methods: A total of 60 female Institute of Cancer Research (ICR) mice aged 6–8 weeks were divided into control group, control group with 0 mgRU486 group (control-0 mgRU486), control group with 2 mgRU486 group (control-2 mgRU486), hyperthyroid pregnant mice with 0 mgRU486 group (hyper-0 mgRU486), hyperthyroid pregnant mice with 2 mgRU486 group (hyper-2 mgRU486), and hyperthyroid pregnant mice with 20 mgRU486 group (hyper-20 mgRU486). In the hyperthyroidism groups, L-thyroxine sodium was intraperitoneally injected every day at 30 μg·kg−1·day−1 until the end of the experiment. On the 7th day of the experiment, free triiodothyronine (FT3), free tetraiodothyroxine (FT4), thyroxine (TT4), and thyroid-stimulating hormone (TSH) levels were tested. The mice in the control groups and those in the experimental groups were paired with the male mice (2:1) on the 10th day of the experiment, and the caging was recorded. On the 8th day of pregnancy (day 8), pregnant mice were subcutaneously injected with mifepristone in different doses and were sacrificed 6 h later. Pregnancy rate and the number of embryos were recorded. Thyroid tissues were observed by hematoxylin and eosin (HE) staining. Serum TSH level was determined by radioimmunoassay. Results: Six hours after injection with mifepristone, serum FT3, FT4, and TT4 levels of pregnant mice were all increased. The increased levels in the mice under hyperthyroidism were different from those in the control groups (P < 0.05). There was no difference in the embryo number and pregnancy rate between the experimental and the control groups; HE staining indicated that there was no significant change in microscopic features before and after mifepristone administration. Conclusion: Serum thyroid hormone level of mice under hyperthyroidism was significantly increased after mifepristone administration. Therefore, mifepristone should be avoided when hyperthyroidism has not been controlled.
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Lin28B expression in reduced uterine perfusion pressure rat model p. 36
Yun Yang, Jia-Po Li, Yue Bian, Gui-Yu Song, Yuan-Yuan Li, Dong-Ying Zheng, Ling Huang, Chong Qiao
Objective: Preeclampsia (PE) is a serious complication of pregnancy. Placental ischemia could be an initiating event, but the molecular mechanisms underlying PE are unclear. Lin28B, a paralog of Lin28 RNA-binding protein, is predominantly expressed in human placenta, and decreased Lin28B expression may play a role in PE by reducing trophoblast invasion. The current study was intended to verify whether Lin28B plays a role in the pathogenesis of PE in rat model for reduced uterine perfusion pressure (RUPP). Methods: We used RUPP rat model. The changes in blood pressure, 24-h urine protein excretion, and fetal development in RUPP rats were recorded and compared to those of normal pregnant (NP) rats. Furthermore, the expression of Lin28B mRNA and protein in placenta was determined using quantitative real-time polymerase chain reaction, Western blotting, and immunohistochemistry. Results: The blood pressure, 24-h urine protein excretion, and embryo absorption rate were significantly increased in RUPP rats on the 20th day of gestational period compared with the NP rats (P < 0.001). However, there was no difference in the weight of placenta in RUPP versus NP rats (P > 0.05). The expression levels of Lin28B mRNA and protein in the placenta of RUPP rats were also significantly decreased in comparison to NP rats (P < 0.001). Conclusion: Our results show that the expression of Lin28B in the placenta of RUPP rats is different from that in NP rats, thus suggesting a role of Lin28B in the pathogenesis of preeclampsia.
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Preliminary research on the effect of linolenic acid on human oocyte maturation p. 42
Ji-Jun Hu, Jia-Xin Li, Xing-Ling Wang, Yi-Chun Guan, Li-Jun Sun
Objective: To investigate the effects of exogenous α-linolenic acid (ALA) on in vitro maturation (IVM) and developmental competence of human oocytes. Methods: Experiment 1 examined the effects of ALA at different concentrations (0 [control], 10, 50, 100, and 200 μmol/L) in the IVM medium on oocyte maturation. Treatment with 50 μmol/L ALA significantly accelerated oocyte maturation (P < 0.05) and resulted in significantly higher mitochondrial DNA (mtDNA) copy number compared to the control. Hence, 50 μmol/L ALA was selected for combination with follicular fluid (FF) to investigate oocyte developmental potential. mtDNA of the matured oocyte was analyzed by real-time polymerase chain reaction. Experiment 2 investigated the effects of FF with optimal ALA concentration (Group A: ALA + FF + IVM medium) or without ALA (Group B: FF + IVM medium) on oocyte maturation, fertilization, 2 pronuclear cleavage, and embryo and blastocyst development. Malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were measured for maturation medium from Group A, Group B, and Group C (control group, IVM medium only). Results: Treatment with 50 μmol/L ALA obviously accelerated oocyte maturation (P < 0.05) and resulted in significantly higher mtDNA copy number (P < 0.05) in the matured oocytes compared to the control (0 μmol/L ALA). Supplementation of 50 μmol/L ALA and FF (Group A) obviously increased the total maturation rate than FF-treated group (Group B) which has higher (P < 0.05) total maturation rate than that of Group C. However, no significant differences were observed in fertilization, embryo availability, and blastocyst production among Group A, B, and C. Treatment with 50 μmol/L ALA decreased the level of MDA (P < 0.05), but had no effect on the activity of SOD in the IVM medium. Conclusions: Our results suggested that the treatment with 50 μmol/L ALA during IVM improves maturation in human oocytes. It is also likely to improve embryo availability and blastocyst formation. This might be associated with the alteration of mtDNA replication (increased mtDNA copy number) and the reduction of oxidative stress (reduced MDA level).
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Influence of growth hormone supplementation in patients with thin endometrium undergoing frozen embryo transfer p. 49
Jun-Yi Yang, He Li, Nan Lu, Lu Li, Xiao-Xi Sun
Objective: To evaluate the effect of recombinant human growth hormone (rhGH) supplementation during hormone-replacement therapy and frozen-thawed embryo transfer (FET) in patients with thin endometrium. Methods: A retrospective research was conducted on 225 patients, who underwent artificial cycle FET in Shanghai, China, between January 2016 and November 2017. Data from 245 FET cycles were analyzed, of which 184 cycles received rhGH (GH group) and 61 did not (control group). Results: Clinical pregnancy and implantation rates were significantly higher in the GH group than those in the control group (64.7% vs. 49.2%, P = 0.032; 44.8% vs. 32.8%, P = 0.019, respectively). After logistic regression analysis, rhGH was considered the only significant variable that influenced clinical pregnancy rate, increasing it by 1.89-fold. On the other hand, the presence of rhGH did not seem to affect the early pregnancy loss. Conclusions: Our results indicated that simultaneous addition of rhGH could improve clinical outcomes of FET in patients with thin endometrium, particularly in patients between 30 and 34 years of age.
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Spectrum of diseases in female children and adolescents admitted to the obstetrics and gynecology hospital, Shanghai, between 2003 and 2017 p. 54
Hai-Xia Huang, Ming-Xing Zhang, Hong-Wei Zhang, Meng Zhang, Yi Yu, Yu Kang, Gui-Ling Li, Cong-Jian Xu
Objective: To investigate the spectrum of diseases in female children and adolescents hospitalized for obstetric and gynecological conditions. Methods: This retrospective study included patients (age ≤19 years) admitted to the Obstetrics and Gynecology Hospital of Fudan University, Shanghai, between January 2003 and December 2017. Their medical records – age, principal diagnosis, and treatment – were reviewed and analyzed. Diagnoses were divided into eight disease classifications. Comparison between groups was analyzed with Student's t-test. Results: This study enrolled 4,628 inpatients. The number of inpatients increased each year, especially in the groups of gynecologic tumors, pregnancy-related conditions, and disorders of the breast. The mean age was 16.33 ± 2.42 years (range: 2–19 years). Of all the inpatients, 41.9% (n = 1,939) were hospitalized for gynecologic tumors, and 85.5% (n = 1,657) of the tumors were benign. Altogether, ovarian cysts (28.66%, n = 1,325) were the most common disease observed in this group. In addition, 32.9% (n = 1,524) of the inpatients were hospitalized for pregnancy, childbirth, and puerperium; within this group, 21.9% (n = 1,014) were hospitalized for artificial abortions. Conclusions: The efforts of all stakeholders toward the implementation of government policies to promote sexual and reproductive health among children and adolescents in China have been effective.
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Effects of perinatal complications on coagulation function in pregnant women p. 60
Wei-Qin Yu, Xiao-Yan Liu, Pei-Min Mao
Objectives: To investigate the effects of various complications on the coagulation function of perinatal women. Methods: A total of 210 pregnant women between November 2016 and June 2017 were recruited. The coagulation function indexes, including prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), and thrombin time (TT), were evaluated in pregnant women with group B Streptococcus (GBS), positive nonstress test (NST), scarred uterus, prenatal fever, macrosomia, gestational hypertension, gestational diabetes, premature rupture of membranes (PROMs), threatened abortion, and oligohydramnios. Results: No significant differences were found in PT, APTT, FIB, and TT in pregnant women with GBS, positive NST, scarred uterus, macrosomia, gestational hypertension, gestational diabetes, PROMs, and oligohydramnios. Moreover, significantly decreased PT and APTT and significantly increased TT were found in pregnant women with prenatal fever compared to normal controls, whereas significantly increased TT was also observed in those with threatened abortion. Conclusions: Among the complications that occur in pregnant women, prenatal fever and threatened abortion have important effects on coagulation function.
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