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1.
Toxicol In Vitro ; 98: 105834, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38657713

ABSTRACT

Triphenyltin chloride (TPTCL) is widely used in various industrial and agricultural applications. This study aimed to elucidate the mechanisms underlying the toxicological effects of TPTCL on oocytes. The obtained findings revealed that TPTCL exposure reduced polar body extrusion (PBE) and induced meiotic arrest. Mechanistically, TPTCL disrupted meiotic spindle assembly and chromosome alignment. Further analysis indicated a significant decrease in p-MAPK expression, and disturbances in the localization of Pericentrin and p-Aurora A in TPTCL exposed oocytes, which suggesting impaired microtubule organizing center (MTOC)function. Moreover, TPTCL exposure enhance microtubule acetylation and microtubule instability. Therefore, the spindle assembly checkpoint (SAC) remained activated, and the activity of the anaphase-promoting complex (APC) was inhibited, thereby preventing oocytes from progressing into the entering anaphase I (AI) stage. TPTCL exposure also augmented the actin filaments in the cytoplasm. Notably, mitochondrial function appeared unaffected by TPTCL, as evidenced indicated by stable mitochondrial membrane potential and ATP content. Furthermore, TPTCL treatment altered H3K27me2, H3K27me3 and H3K9me3 levels, suggesting changes in epigenetic modifications in oocytes. Taken together, our results suggest that TPTCL disrupts cytoskeleton assembly, continuously activates SAC, inhibits APC activity, and blocks meiotic progression, ultimately impair oocyte maturation.


Subject(s)
Cytoskeleton , Meiosis , Oocytes , Organotin Compounds , Animals , Oocytes/drug effects , Meiosis/drug effects , Female , Cytoskeleton/drug effects , Organotin Compounds/toxicity , Mice , Mice, Inbred ICR , Cell Cycle/drug effects
2.
Ecotoxicol Environ Saf ; 276: 116312, 2024 May.
Article in English | MEDLINE | ID: mdl-38608383

ABSTRACT

The use of bisphenol A (BPA) has been restricted due to its endocrine-disrupting effects. As a widely used alternative to BPA today, environmental levels of bisphenol Z (BPZ) continue to rise and accumulate in humans. Oocyte quality is critical for a successful pregnancy. Nevertheless, the toxic impacts of BPZ on the maturation of mammalian oocytes remain unexplored. Therefore, the impacts of BPZ and BPA on oocyte meiotic maturation were compared in an in vitro mouse oocyte culture model. Exposure to 150 µM of both BPZ and BPA disrupted the assembly of the meiotic spindle and the alignment of chromosomes, and BPZ exerted stronger toxicological effects than BPA. Furthermore, BPZ resulted in aberrant expression of F-actin, preventing the formation of the actin cap. Mechanistically, BPZ exposure disrupted the mitochondrial localization pattern, reduced mitochondrial membrane potential and ATP content, leading to impaired mitochondrial function. Further studies revealed that BPZ exposure resulted in oxidative stress and altered expression of genes associated with anti-oxidative stress. Moreover, BPZ induced severe DNA damage and triggered early apoptosis in oocytes, accompanied by impaired lysosomal function. Overall, the data in this study suggest that BPZ is not a safe alternative to BPA. BPZ can trigger early apoptosis by affecting mitochondrial function and causing oxidative stress and DNA damage in oocytes. These processes disrupt cytoskeletal assembly, arrest the cell cycle, and ultimately inhibit oocyte meiotic maturation.


Subject(s)
Benzhydryl Compounds , DNA Damage , Endocrine Disruptors , Meiosis , Mitochondria , Oocytes , Oxidative Stress , Phenols , Animals , Phenols/toxicity , Oocytes/drug effects , Benzhydryl Compounds/toxicity , Meiosis/drug effects , Mitochondria/drug effects , Mice , Oxidative Stress/drug effects , Female , Endocrine Disruptors/toxicity , Apoptosis/drug effects , Membrane Potential, Mitochondrial/drug effects , Actins/metabolism
3.
Aging (Albany NY) ; 16(5): 4920-4942, 2024 Mar 08.
Article in English | MEDLINE | ID: mdl-38461430

ABSTRACT

OBJECTIVE: Studies have firmly established the pivotal role of the immunogenic cell death (ICD) in the development of tumors. This study seeks to develop a risk model related to ICD to predict the prognosis of patients with endometrial carcinoma (EC). MATERIALS AND METHODS: RNA-seq data of EC retrieved from TCGA database were analyzed using R software. We determined clusters based on ICD-related genes (ICDRGs) expression levels. Cox and LASSO analyses were further used to build the prediction model, and its accuracy was evaluated in the train and validation sets. Finally, in vitro and in vivo experiments were conducted to confirm the impact of the high-risk gene IFNA2 on EC. RESULTS: Patients were sorted into two ICD clusters, with survival analysis revealing divergent prognoses between the clusters. The Cox regression analysis identified prognostic risk genes, and the LASSO analysis constructed a model based on 9 of these genes. Notably, this model displayed excellent predictive accuracy when validated. Finally, increased IFNA2 levels led to decreased vitality, proliferation, and invasiveness in vitro. IFNA2 also has significant tumor inhibiting effect in vivo. CONCLUSIONS: The ICD-related model can accurately predict the prognosis of patients with EC, and IFNA2 may be a potential treatment target.


Subject(s)
Endometrial Neoplasms , Immunogenic Cell Death , Humans , Female , Prognosis , Endometrial Neoplasms/genetics , Cell Movement , Databases, Factual , Tumor Microenvironment
4.
Nat Biotechnol ; 2024 Feb 09.
Article in English | MEDLINE | ID: mdl-38336903

ABSTRACT

Dynamic 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) modifications to DNA regulate gene expression in a cell-type-specific manner and are associated with various biological processes, but the two modalities have not yet been measured simultaneously from the same genome at the single-cell level. Here we present SIMPLE-seq, a scalable, base resolution method for joint analysis of 5mC and 5hmC from thousands of single cells. Based on orthogonal labeling and recording of 'C-to-T' mutational signals from 5mC and 5hmC sites, SIMPLE-seq detects these two modifications from the same molecules in single cells and enables unbiased DNA methylation dynamics analysis of heterogeneous biological samples. We applied this method to mouse embryonic stem cells, human peripheral blood mononuclear cells and mouse brain to give joint epigenome maps at single-cell and single-molecule resolution. Integrated analysis of these two cytosine modifications reveals distinct epigenetic patterns associated with divergent regulatory programs in different cell types as well as cell states.

5.
Hum Fertil (Camb) ; 26(6): 1503-1510, 2023 Dec.
Article in English | MEDLINE | ID: mdl-38248811

ABSTRACT

The influence of cola intake on birth outcomes is unclear. This study sought to describe and compare the associations between cola intake and adverse birth outcomes among women following assisted reproductive technology (ART) and women spontaneously conceived (SC). Participants (736 ART women and 1,270 SC women) were from the Chinese National Birth Cohort collected in Anhui province. Cola intake was assessed by self-reported questionnaires at each trimester. Outcome measures including preterm birth (PTB) and low birth weight (LBW) were extracted from medical records. The association between cola intake during pregnancy and PTB was found using multivariable log-binomial regression in combined ART and SC women. Separately, for ART women, cola intake during pregnancy increased the risk of PTB (risk ratios were 2.10, 1.65, and 1.81 for all three trimesters, respectively, all p < 0.05), and cola intake in the 1st trimester increased the risk of LWB (risk ratio 2.58, 95% confidence interval 1.29 to 5.16). Cola intake during pregnancy was not associated with PTB or LBW for SC women. Our findings indicate a detrimental effect of cola intake during pregnancy on birth outcomes for ART women. Thus, avoidance of cola intake should be counselled by medical doctors in women prescribed with ART treatment.


Subject(s)
Carbonated Beverages , Cola , Pregnancy , Premature Birth , Reproductive Techniques, Assisted , Female , Humans , Infant, Newborn , Pregnancy/drug effects , Asian People , Cohort Studies , Premature Birth/epidemiology , Premature Birth/etiology , Reproductive Techniques, Assisted/adverse effects , Carbonated Beverages/adverse effects , Cola/adverse effects , Pregnancy Outcome
6.
J Obstet Gynaecol Res ; 48(7): 1501-1512, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35445497

ABSTRACT

AIM: This review aims to summarize the research focused upon the functions of nuclear hormone receptor 4A (NR4A) in the human reproductive system. The research questions addressed are to decipher what role the NR4A subfamily plays in the regulation of the human reproductive system and effects upon fertility issues through regulation of the expression of the NR4A subfamily. METHODS: The electronic database PubMed was searched for studies published before November 2021. Keywords included "NR4A," "trophoblast," "decidualization," "folliculogenesis," "estrogen," "pregnancy," "Leydig cells," "fertility," and "reproductive." Relevant references from retrieved manuscripts and review articles were also searched manually. RESULTS: NR4A subfamily are involved in trophoblast differentiation, endometrial decidualization, embryo adhesion, secretion of related hormones, and regulation of spontaneous term labor. Besides, many studies have provided strong evidence that they play critical roles in spermatogenesis. Furthermore, Multiple mechanisms can affect the expression of NR4As. Broadly, NR4A family receptors affect the human reproductive system in multiple ways. CONCLUSIONS: Further research is needed to specifically dissect the functions and regulatory mechanisms of these receptors and their pharmaceutical antagonists and agonists. The connection between the NR4A subfamily and a variety of reproductive disorders needs to be proven experimentally such that further examination of human tissue is required to assess the role of these receptors in human reproductive diseases.


Subject(s)
Gene Expression Regulation , Nuclear Receptor Subfamily 4, Group A, Member 1 , Endometrium/metabolism , Female , Humans , Male , Nuclear Receptor Subfamily 4, Group A, Member 1/genetics , Nuclear Receptor Subfamily 4, Group A, Member 1/metabolism , Signal Transduction , Trophoblasts/metabolism
7.
Front Genet ; 12: 746082, 2021.
Article in English | MEDLINE | ID: mdl-34925444

ABSTRACT

Recurrent pregnancy loss (RPL) is a common reproductive problem affecting around 5% of couples worldwide. At present, about half of RPL cases remained unexplained. Previous studies have suggested an important role for genetic determinants in the etiology of RPL. Here, we performed whole-exome sequencing (WES) analysis on 100 unrelated Han Chinese women with a history of two or more spontaneous abortions. We identified 6736 rare deleterious nonsynonymous variants across all patients. To focus on possible candidate genes, we generated a list of 95 highly relevant genes that were functionally associated with miscarriage according to human and mouse model studies, and found 35 heterozygous variants of 28 RPL-associated genes in 32 patients. Four genes (FOXA2, FGA, F13A1, and KHDC3L) were identified as being strong candidates. The FOXA2 nonsense variant was for the first time reported here in women with RPL. FOXA2 knockdown in HEK-293T cells significantly diminished the mRNA and protein expression levels of LIF, a pivotal factor for maternal receptivity and blastocyst implantation. The other genes, with 29 variants, were involved in angiogenesis, the immune response and inflammation, cell growth and proliferation, which are functionally important processes for implantation and pregnancy. Our study identified several potential causal genetic variants in women with RPL by WES, highlighting the important role of genes controlling coagulation, confirming the pathogenic role of KHDC3L and identifying FOXA2 as a newly identified causal gene in women with RPL.

8.
Eur J Pharmacol ; 890: 173659, 2021 Jan 05.
Article in English | MEDLINE | ID: mdl-33131637

ABSTRACT

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen of 2019 novel coronavirus disease (COVID-19), is currently spreading around the world. The WHO declared on January 31 that the outbreak of SARS-CoV-2 was a public health emergency. SARS-Cov-2 is a member of highly pathogenic coronavirus group that also consists of severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV). Although respiratory tract lesions were regarded as main manifestation of SARS-Cov-2 infection, gastrointestinal lesions were also reported. Similarly, patients with SARS-CoV and MERS-CoV were also observed. Common gastrointestinal symptoms of patients mainly included diarrhea, vomiting and abdominal pain. Gastrointestinal lesions could be used as basis for early diagnosis of patients, and at the same time, controlling gastrointestinal lesions better facilitated to cut off the route of fecal-oral transmission. Hence, this review summarizes the characteristics and mechanism of gastrointestinal lesions caused by three highly pathogenic human coronavirus infections including SARS-CoV, MERS-CoV, as well as SARS-CoV-2. Furthermore, it is expected to gain experience from gastrointestinal lesions caused by SARS-CoV and MERS-CoV infections in order to be able to better relieve SARS-CoV-2 epidemic. Targetin gut microbiota to regulate the process of SARS-CoV-2 infection should be a concern. Especially, the application of nanotechnology may provide help for further controlling COVID-19.


Subject(s)
Coronavirus Infections/complications , Gastrointestinal Diseases/etiology , Middle East Respiratory Syndrome Coronavirus , SARS-CoV-2 , Severe acute respiratory syndrome-related coronavirus , Animals , Humans
9.
Life Sci ; 250: 117543, 2020 Jun 01.
Article in English | MEDLINE | ID: mdl-32169518

ABSTRACT

AIMS: HMGB1 has been reported to play a crucial role in the physiological and pathophysiological responses during pregnancy. However, it is still unknown whether excessively expressed HMGB1 at the maternal-fetal interface related to Unexplained Recurrent Spontaneous Abortion (URSA). This study was designed to investigate the local capability of HMGB1 in the pathology of URSA, determined the distributions and characteristics of HMGB1, its receptors (RAGE/TLR2/TLR4) and important signaling molecule NF-κB p65 expression at the maternal-fetal interface,as well as compared the differences of HMGB1 expression between the URSA group, control group and aspirin treatment group. MATERIAL AND METHODS: H&E staining, Western blot analysis, immunofluorescence assay and immunohistochemical staining were applied to determine the effect of HMGB1 and its receptors at the maternal-fetal interface. ELISA was utilized to detect the concentration of HMGB1 in plasma. KEY FINDINGS: Our study demonstrated that the activation of the HMGB1-RAGE/TLR2/TLR4-NF-κB pathway at the maternal-fetal interface may have occurred in the URSA group. HMGB1 concentration in plasma was higher in the URSA group than the control group. Furthermore, the levels of HMGB1 of subjects with URSA could be reduced by administrating low doses of aspirin (ASPL). SIGNIFICANCE: This is the first report indicating the roles of HMGB1 at the maternal-fetal interface of URSA patients and broadening the horizons for clinical treatment of URSA. HMGB1-RAGE/TLR2/TLR4-NF-κB signaling pathway may be activated at the maternal-fetal interface in URSA and account for its pathogenesis. HMGB1 have the potential to be promising biomarkers in prevention and therapy of URSA.


Subject(s)
Abortion, Habitual/metabolism , Abortion, Spontaneous/metabolism , HMGB1 Protein/metabolism , Signal Transduction , Antigens, Neoplasm/metabolism , Aspirin/administration & dosage , Female , Fetus , Gene Expression Regulation , Humans , Leukocyte Common Antigens/metabolism , Maternal-Fetal Exchange , Mitogen-Activated Protein Kinases/metabolism , Pregnancy , Toll-Like Receptor 2/metabolism , Toll-Like Receptor 4/metabolism , Transcription Factor RelA/metabolism , Up-Regulation
10.
Am J Pathol ; 189(1): 124-131, 2019 01.
Article in English | MEDLINE | ID: mdl-30339840

ABSTRACT

Recurrent pregnancy loss (RPL) is a major concern for women's reproductive health. Several studies have proved that genetics is a major factor leading to unexplained RPL, but the maternal pathogenic genes involved in RPL remain largely unknown. A consanguineous family, including the parents who were cousins and their three daughters who had been diagnosed as having nonsyndromic unexplained RPL, was recruited in this study. A rare homozygous variant in calcyphosine (CAPS; ENST00000588776: c.377delC, p.Leu127Trpfs) might be the potential candidate variant for this RPL family through whole-exome sequencing. Sanger sequencing confirmed that the three affected sisters carried the homozygous p.Leu127Trpfs, whereas their parents carried the heterozygous p.Leu127Trpfs. CAPS encodes a Ca2+-binding protein and may play a role in the regulation of Ca2+ transport. Although the precise underlying mechanisms remain unclear, the previous study suggested that they may be involved in cross talk between Ca2+ signaling and cAMP-protein kinase A pathways, which are crucial to embryo implantation and pregnancy maintenance. Knockdown of CAPS expression might promote the expression of secreted phosphoprotein 1 and matrix metalloproteinase 9, and the release of prostaglandin E2, which all played important roles in embryo implantation and early pregnancy maintenance. These results indicated that the autosomal recessive homozygous mutation, p.Leu127Trpfs, in CAPS might be a maternal effect causative mutation of RPL pathogenesis.


Subject(s)
Abortion, Spontaneous , Base Sequence , Calcium-Binding Proteins , Genes, Recessive , Sequence Deletion , Abortion, Spontaneous/genetics , Abortion, Spontaneous/metabolism , Abortion, Spontaneous/pathology , Adult , Calcium Signaling/genetics , Calcium-Binding Proteins/genetics , Calcium-Binding Proteins/metabolism , Cyclic AMP-Dependent Protein Kinases/genetics , Cyclic AMP-Dependent Protein Kinases/metabolism , Dinoprostone/genetics , Dinoprostone/metabolism , Embryo Implantation/genetics , Female , Humans , Matrix Metalloproteinase 9/genetics , Matrix Metalloproteinase 9/metabolism , Osteopontin/genetics , Osteopontin/metabolism , Pregnancy , Exome Sequencing
11.
J Cell Biochem ; 120(5): 8291-8299, 2019 May.
Article in English | MEDLINE | ID: mdl-30548667

ABSTRACT

STOX1 is a transcription factor that is implicated in the high prevalence of human gestational diseases. It has been studied in various types of gestational diseases using different molecular and cellular biological technologies. However, the effect and detailed mechanism of storkhead box 1 (STOX1) in recurrent spontaneous abortion (RSA) remain unknown. This study aimed to explore the effect and detailed mechanism of STOX1 in human trophoblast cells. The result showed that downregulation of STOX1 by short hairpin RNA (shRNA) led to a decrease in proliferation and migration in HTR-8/SVneo cells, while it induced the apoptosis of HTR-8/SVneo cells. Moreover, the result showed that trophoblast cells expressed lower levels of pAKT and p85 subunits after treatment with STOX1 shRNA when compared with control. However, overexpression of STOX1 obviously increased the pAKT and p85 protein expressions. Transfection of pcDNA-AKT plasmid increased cell proliferation and migration in HTR-8/SVneo cells while suppressed the apoptosis of HTR-8/SVneo cells. Furthermore, inhibition of the PI3K/Akt pathway by a specific inhibitor promoted cell apoptosis and aggravatedly suppressed cell proliferation and migration of HTR-8/SVneo cells. On the other hand, upregulation of the PI3K/Akt pathway could increase the relative expression level of Bcl-2 and decrease the relative expression levels of Bax and Bim, while inhibition of the PI3K/Akt pathway led to adverse results. Our results demonstrated that inhibition of STOX1 could suppress trophoblast cell proliferation and migration, while promote apoptosis through inhibiting the PI3K/Akt signaling pathway. These findings might provide a new fundamental mechanism for regulating RSA and could be used to prevent and treat RSA in clinic.

12.
Biomed Res Int ; 2018: 9846274, 2018.
Article in English | MEDLINE | ID: mdl-30155486

ABSTRACT

Selecting excellent oocytes is required to improve the outcomes of in vitro fertilization (IVF). Cumulus cells (CCs) are an integral part of the oocyte maturation process. Therefore, we sought to identify differentially expressed genes in CCs to assess oocyte quality and embryo development potential. We divided the participants' embryos into the high-quality embryo group and low-quality embryo group by the information including age, body mass index, and the levels of luteinizing hormone, follicle-stimulating hormone, estradiol, and progesterone. We analyzed a total of 7 CC samples after the quality control in RNA sequencing. We found that 2499 genes were unregulated and 5739 genes were downregulated in high-quality embryo group compared to the low-quality embryo group (Padj < 0.05). Interestingly, MSTN, CTGF, NDUFA1, VCAN, SCD5, and STAR were significantly associated with the quality of embryo. In accordance with the results of RNA sequencing, the association of the expression levels of MSTN, CTGF, NDUFA1, VCAN, SCD5, and STAR with the embryo quality was verified by quantitative reverse-transcription polymerase chain reaction (RT-qPCR) in 50 CC samples. Despite the small sample size and lack of validation in animal models, our study supports the fact that differential gene expression profile of human CCs, including MSTN, CTGF, NDUFA1, VCAN, SCD5, and STAR, can serve as potential indicator for embryo quality.


Subject(s)
Cumulus Cells/metabolism , Fertilization in Vitro , Oocytes , Sequence Analysis, RNA , Animals , China , Female , Humans , Male , Semen Analysis , Sperm Motility , Transcriptome
13.
Clin Appl Thromb Hemost ; 24(6): 894-900, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29363996

ABSTRACT

Mutations of hemostasis/coagulation-related genes have been speculated to cause recurrent spontaneous abortion (RSA). This study investigated the genetic association between the polymorphisms of factor V (F5), factor II (F2), antithrombin (SERPINC1), protein C (PROC), protein S (PROS1), protein Z (PROZ), factor XIII (F13A1), and carboxypeptidase B2 (CPB2) genes and RSA. The 426 patients with RSA and 444 controls were recruited in this study, and single-nucleotide polymorphisms (SNPs) were analyzed by using SNPscan technology. Genotype and allele frequencies of rs3136520 in F2, rs3024731 in PROZ, and rs1050782 in F13A1 showed statistically significant differences between the 2 groups. TT genotype of rs3136520 ( P = .031, odds ratio [OR] = 0.986, 95% confidence interval [CI] = 0.976-0.997) and AA genotype of rs2069906 in PROC ( P = .021, OR = 0.114, 95% CI = 0.014-0.902) in their recessive models and AG + GG variants of rs1050782 ( P = .007, OR = 0.681, 95% CI = 0.516-0.899) in the dominant model might be associated with the reduced risk of RSA. AT + TT variants of rs3024731 ( P = .010, OR = 1.479, 95% CI = 1.098-1.994) may increase disease susceptibility in dominant model. Haplotype analysis of rs3024731 and rs3024735 in PROZ displayed that the AA and TG haplotype were inclined to decrease and increase the risk of RSA, respectively. These results suggested that rs3136520, rs2069906, rs3024731, and rs1050782 may have a significant association with the genetic susceptibility of RSA in Chinese Han women.


Subject(s)
Abortion, Habitual/genetics , Blood Proteins/genetics , Factor XIII/genetics , Models, Genetic , Polymorphism, Single Nucleotide , Protein C/genetics , Prothrombin/genetics , Abortion, Habitual/epidemiology , Abortion, Habitual/ethnology , Asian People , China/epidemiology , Female , Haplotypes , Humans
14.
J Assist Reprod Genet ; 34(1): 125-129, 2017 Jan.
Article in English | MEDLINE | ID: mdl-27722936

ABSTRACT

PURPOSE: The study aims to investigate the genetic association between paired box gene 2 (PAX2) and mullerian duct anomalies (MDA) in Chinese Han females. METHODS: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to identify the genotypes of three tag single nucleotide polymorphisms (SNPs) in PAX2 in 362 MDA cases and 406 controls. RESULTS: We found that one tag SNP (rs12266644) of PAX2 was associated with susceptibility to MDA. The genotype distributions of the SNP rs12266644 have a statistically significant difference in the MDA patients and controls with a p value = 0.008. In the dominant model, we also observed that the GT + TT genotype increased the risk for MDA (p = 0.015, OR = 1.637, 95 % CI = 1.096-2.443). CONCLUSION: The polymorphism rs12266644 of PAX2 might be a risk factor for MDA in Chinese Han females.


Subject(s)
Genetic Association Studies , Genital Diseases, Female/genetics , Mullerian Ducts/pathology , PAX2 Transcription Factor/genetics , Adult , Alleles , Asian People , China , Female , Genetic Predisposition to Disease , Genital Diseases, Female/pathology , Genotype , Humans , Middle Aged , Polymorphism, Single Nucleotide , Risk Factors , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
15.
Sci Rep ; 6: 39638, 2016 12 22.
Article in English | MEDLINE | ID: mdl-28004769

ABSTRACT

Polycystic ovary syndrome (PCOS) is a common frequent endocrine disorder among women of reproductive age. Although assisted reproductive techniques (ARTs) are used to address subfertility in PCOS women, their effectiveness is not clear. Our aim was to compare transcriptomic profiles of oocytes and cumulus cells (CCs) between women with and without PCOS, and assess the effectiveness of ARTs in treating PCOS patients. We collected oocytes and CCs from 16 patients with and without PCOS patients to categorize them into 6 groups according to oocyte nuclear maturation. Transcriptional gene expression of oocyte and CCs was determined via single-cell RNA sequencing. The ratio of fertilization and cleavage was higher in PCOS patients than in non-PCOS patients undergoing ARTs, and there was no difference in the number of high-quality embryos between the groups. Differentially expressed genes including PPP2R1A, PDGFRA, EGFR, GJA1, PTGS2, TNFAIP6, TGF-ß1, CAV1, INHBB et al. were investigated as potential causes of PCOS oocytes and CCs disorder at early stages, but their expression returned to the normal level at the metaphase II (MII) stage via ARTs. In conclusion, ARTs can improve the quality of cumulus-oocyte complex (COC) and increase the ratio of fertilization and cleavage in PCOS women.


Subject(s)
Cumulus Cells/metabolism , Oocytes/metabolism , Polycystic Ovary Syndrome/metabolism , Single-Cell Analysis , Transcriptome , Adult , Body Mass Index , Cumulus Cells/cytology , Female , Gap Junctions/pathology , Gene Expression Profiling , Gene Expression Regulation , Humans , Metaphase , Oocytes/cytology , Oogenesis , Oxidative Stress , Reproductive Techniques, Assisted , Sequence Analysis, RNA , Signal Transduction
16.
Reprod Biomed Online ; 32(4): 420-6, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26856455

ABSTRACT

Dapper antagonist of catenin-1 (DACT1) plays an important role in embryogenesis and organogenesis of the female reproductive tract in mouse models. The aim of this study was to investigate the association between DACT1 mutations and human Müllerian duct anomalies (MDA). One hundred clinically well-defined Chinese Han patients with MDA and 200 healthy controls were recruited in this study. All four exons coding for DACT1 were amplified and sequenced. A missense mutation (c.G1084A, p.V362M) was identified in a patient who had a didelphic uterus and was absent from the control group. This variant changed the hydrophilicity of the amino acid residue and was predicted to be deleterious to the structure and function of DACT1 protein. The data indicate that the p.V362M mutation of DACT1 may be an underlying cause of MDA.


Subject(s)
Adaptor Proteins, Signal Transducing/genetics , Mullerian Ducts/abnormalities , Nuclear Proteins/genetics , Adult , China , DNA Mutational Analysis , Female , Humans , Mullerian Ducts/embryology , Mutation, Missense , Wnt Signaling Pathway
17.
Syst Biol Reprod Med ; 61(4): 205-10, 2015.
Article in English | MEDLINE | ID: mdl-25848831

ABSTRACT

Recurrent spontaneous abortion (RSA) is a multi-factor disease. The mammalian target of the the rapamycin (MTOR) gene has been reported to be involved in mouse embryo development and regulates the proliferation of embryonic stem cells. Our study explored the relationship between the single nucleotide polymorphism (SNP) rs17027478 in the promoter region of MTOR gene and the development of RSA. A total of 306 patients with RSA and 127 healthy females as the controls were recruited in the case-control study. The predesigned TaqMan SNP Genotyping Assay was adopted to analyze the association between rs17027478 and the development of RSA. Quantitative real-time reverse transcription polymerase chain reaction and luciferase reporter assays were conducted to analyze the function of the variant. It was found that a significant association exists between the variant and the risk of RSA among the patients who experienced no less than three spontaneous abortions (p = 0.043). However, the significant difference disappeared among the total samples (p = 0.524). Furthermore, we observed lower MTOR mRNA levels in the blood of RSA patients compared with healthy females (p = 0.020). The luciferase reporter assay showed that the rs17027478A allele significantly reduced the luciferase activity (p = 0.029). The results demonstrated that the variant rs17027478 in the promoter region of MTOR might be a good candidate responsible for the pathogenesis of RSA. Abbreviations RSA recurrent spontaneous abortion MTOR mammalian target of rapamycin SNP single nucleotide polymorphism qRT-PCR quantitative real-time polymerase chain reaction URSA unexplained recurrent spontaneous abortion mTORC1 mTOR complex 1 ESC embryonic stem cells HKE-293 human embryonic kidney 293 cells HWE Hardy-Weinberg equilibrium ANOVA one-way analysis of variance.


Subject(s)
Abortion, Habitual/genetics , Polymorphism, Single Nucleotide , TOR Serine-Threonine Kinases/genetics , Case-Control Studies , China , Female , Humans , Pregnancy , RNA, Messenger/genetics
18.
Reprod Sci ; 22(8): 1008-14, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25670720

ABSTRACT

The close relationship between cumulus cells and oocyte indicates that the analysis of cumulus gene expression is a potential noninvasive method to aid embryo selection and in vitro fertilization outcome. Long noncoding RNAs (LncRNAs) could regulate essential pathways that contribute to human oocyte maturation, fertilization, and embryo development, which indicates that lncRNA would be valuable biomarkers. In our previous study, AK124742 is a newly detected lncRNA that was identified as being natural antisense to PSMD6, but its role in oocyte and embryo development is still not elucidated and needs to be investigated. Here, the expression of AK124742 and PSMD6 was measured in 40 pairs of cumulus cells from oocytes that result in high-quality embryos (HCCs) and from oocytes that result in poor-quality embryos (PCCs) by real-time quantitative reverse transcriptase polymerase chain reaction. The predictive value of AK124742 and PSMD6 was evaluated using a receiver-operating characteristic (ROC) curve. Notably, elevated expression levels of AK124742 and PSMD6 were observed in HCCs compared to PCCs (72.5% and 62.5%, respectively; P < .01). Expression of AK124742 was potentially positively associated with the PSMD6 levels. The relative expression levels of AK124742 and PSMD6 in the pregnancy group were significantly higher than those in the nonpregnancy group (P < .01).The area under the ROC curve of AK124742 was 0.78 (95% confidence interval: 0.64-0.93). In conclusion, AK124742 and PSMD6 as a new lncRNA-messenger RNA gene pair in human cumulus cells may be considered as potential biomarkers to aid embryo selection.


Subject(s)
Blastocyst/metabolism , Cumulus Cells/metabolism , Fertilization in Vitro , In Vitro Oocyte Maturation Techniques , Oocytes/metabolism , Proteasome Endopeptidase Complex/genetics , RNA, Long Noncoding/genetics , RNA, Messenger/genetics , Area Under Curve , Blastocyst/pathology , Blastomeres/metabolism , Cells, Cultured , Embryo Culture Techniques , Female , Gene Expression Regulation, Developmental , Genetic Markers , Humans , Predictive Value of Tests , Pregnancy , Pregnancy Rate , Proteasome Endopeptidase Complex/metabolism , RNA, Long Noncoding/metabolism , RNA, Messenger/metabolism , ROC Curve , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Single Embryo Transfer , Treatment Outcome
19.
Reprod Biomed Online ; 29(4): 445-51, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25164168

ABSTRACT

In this study, the clinical significance of first-trimester intrauterine haematomas (IUH) detected in pregnancies achieved by IVF-embryo transfer (IVF-ET) was evaluated. A retrospective case-control study was designed to compare obstetric and perinatal outcomes of 350 pregnancies with IUH and 350 matched controls without IUH. The incidence of first-trimester IUH detected in the IVF-ET pregnancies was 13.5%. In women who delivered after 28 weeks' gestation, the incidence of gestational hypertension (OR 2.6; 95% CI 1.5 to 4.6), preeclampsia (OR 2.8; 95% CI 1.5 to 5.0) and postpartum haemorrhage (OR 3.1; 95% CI 1.8 to 5.3) was significantly higher in the IUH group. Compared with controls, placenta previa (OR, 8.7 95%; CI 3.4 to 22.2) and oligohydramninos (OR 5.8; 95% CI 2.4 to 14.0) were more common in the IUH group. The incidence of preterm delivery (<37 weeks' gestation) was significantly higher in the IUH group (OR 2.1; 95% CI 1.4 to 3.0), although the incidence of preterm delivery before 34 weeks' gestation was not. No differences were observed in the incidence of gestational diabetes mellitus, premature rupture of membranes and low birth weight. The presence of first-trimester IUH in IVF-ET pregnancies was associated with a higher risk of several pregnancy complications.


Subject(s)
Embryo Transfer/adverse effects , Hematoma/physiopathology , Hypertension, Pregnancy-Induced/etiology , Postpartum Hemorrhage/etiology , Pre-Eclampsia/etiology , Pregnancy Complications/physiopathology , Uterine Diseases/physiopathology , Adult , Case-Control Studies , China/epidemiology , Female , Fertilization in Vitro/adverse effects , Hematoma/diagnostic imaging , Hematoma/epidemiology , Hematoma/etiology , Humans , Hypertension, Pregnancy-Induced/epidemiology , Incidence , Infertility, Female/complications , Infertility, Female/therapy , Infertility, Male/therapy , Male , Oligohydramnios/epidemiology , Oligohydramnios/etiology , Placenta Previa/epidemiology , Placenta Previa/etiology , Postpartum Hemorrhage/epidemiology , Pre-Eclampsia/epidemiology , Pregnancy , Pregnancy Complications/diagnostic imaging , Pregnancy Complications/epidemiology , Pregnancy Complications/etiology , Pregnancy Trimester, First , Premature Birth/epidemiology , Premature Birth/etiology , Retrospective Studies , Ultrasonography, Prenatal , Uterine Diseases/diagnostic imaging , Uterine Diseases/epidemiology , Uterine Diseases/etiology
20.
J Assist Reprod Genet ; 31(3): 379-82, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24390679

ABSTRACT

OBJECTIVE: Recurrent spontaneous abortion (RSA) is a multifactor and distressing disease. There are still approximately half of the RSA patients with cause not being identified to date. Accumulating studies have confirmed that genetic polymorphisms in glutathione S-transferases (GSTs) were associated with the risk of recurrent spontaneous abortion. In this study, we aimed to investigate the relationship between the polymorphism of GSTA1, which is GSTA1 -69C/T (rs3957357), and the development of recurrent spontaneous abortion. METHODS: A case-control study of 127 cases with RSA and 112 ethnic and age matched women as controls was conducted. And measurement of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) was performed to genotype all of samples in order to analyze the association between GSTA1 -69C/T (rs3957357) and the risk of RSA. RESULTS: We found that the frequencies of genotypes between cases and controls have no significant difference (P = 0.908) and GSTA1 mutant allele GSTA1 -69 T was present at a frequency of 0.122 in case group, while in controls the frequency was 0.125 (P = 0.922). CONCLUSION: The polymorphism of GSTA1 (rs3957357) may not be associated with the risk of recurrent spontaneous abortion in Chinese Han population.


Subject(s)
Abortion, Habitual/genetics , Genetic Association Studies , Genetic Predisposition to Disease , Glutathione Transferase/genetics , Abortion, Habitual/pathology , Adult , Alleles , Case-Control Studies , China , Female , Gene Frequency , Genotype , Humans , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Pregnancy , Young Adult
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