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OBJECTIVE: Preeclampsia (PE) is a common but serious pregnancy complication that adversely affects both maternal and fetal health. However, the mechanisms of its pathogenesis remain unclear, and effective biomarkers for early diagnosis are still lacking. METHODS: In this retrospective study, comprehensive bioinformatic analysis and logistic regression analysis were used to compare profiles of 48 serum cytokines in 27 PE patients with those in 41 normotensive pregnant subjects. RESULTS: The results revealed that serum cytokine profiles accumulated to different levels between the two groups, which had significant correlations with the clinical features of PE. Nine cytokines with high discriminatory capacity for diagnosising PE (AUC ≥ 0.7) were selected for inclusion in a multivariate logistic regression model for PE and calculated as a probability diagnostic formula. This model constructed from the panel of nine cytokines had better diagnostic performance than any individual cytokine (AUC = 0.97, 95% CI 0.94-1.00, P < 0.0001), with a sensitivity of 96.30% and a specificity of 90.24%. CONCLUSIONS: The set of cytokine profiles and risk assessment model described here can serve as a basis for developing early clinical diagnostic and therapeutic strategies for PE.
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Pré-Eclâmpsia , Gravidez , Feminino , Humanos , Pré-Eclâmpsia/diagnóstico , Pré-Eclâmpsia/etiologia , Estudos Retrospectivos , Biomarcadores , CitocinasRESUMO
BACKGROUND Preeclampsia (PE) is a serious pregnancy disorder associated with immune tolerance imbalance. The etiology of preeclampsia has not been fully elucidated. The aim of this study was to clarify the possible role of the lymphocyte activation gene 3 (LAG-3)/fibrinogen-like protein 1 (FGL-1) signaling pathway in the immune imbalance of early-onset PE. MATERIAL AND METHODS We enrolled 34 women with early-onset PE and 34 age-matched normal pregnancies (NPs). Flow cytometry was performed to determine the expression of LAG-3 on peripheral T cell subsets (CD3+, CD4+, and CD8+ T cells). We measured LAG-3 expression on decidual T cells to determine whether there was a difference in the expression of LAG-3 between decidual and peripheral T cells. Maternal plasma levels of FGL-1 were measured by ELISA. RESULTS There was no significant difference in LAG-3 expression on peripheral CD3+ T cells between NP and early-onset PE. Compared to NP, the significant decrease expression of LAG-3 by peripheral CD4+ and CD8+T cells was found in early-onset PE. The LAG-3 expression was higher on decidual T cells than peripheral counterparts in all pregnancies. The plasma level of FGL-1 was significantly elevated in early-onset PE compared with NP. CONCLUSIONS Abnormal expression of LAG-3/FGL-1 signaling pathway may be associated with immune activation of effector T cells and impaired immune tolerance in early-onset PE.
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Pré-Eclâmpsia , Antígenos CD/metabolismo , Linfócitos T CD8-Positivos , Feminino , Fibrinogênio/metabolismo , Humanos , Gravidez , Transdução de Sinais , Subpopulações de Linfócitos T , Proteína do Gene 3 de Ativação de LinfócitosRESUMO
OBJECTIVE: To assess the value of combined fetal karyotyping and chromosomal microarray analysis (CMA) for the verification of high-risk pregnancy signaled by noninvasive prenatal screening (NIPS) based on high-throughput sequencing. METHODS: One hundred and fifty-one pregnant women with high risks for aneuploidies of chromosomes 13, 18, 21, X and Y or pathological copy number variations (CNVs) by NIPS were subjected to amniocytic karyotyping and CMA analysis. RESULTS: One hundred and forty-two women were found to have a high risk for fetal chromosomal aneuploidies, which included 83 cases of trisomy 21, 17 cases of trisomy 18, 2 cases of trisomy 13, and 40 cases of sex chromosome aneuploidies. Amniocytic karyotyping and CMA analysis has confirmed 81 cases of trisomy 21, 15 cases of trisomy 18, 10 cases of 47,XXY, 4 cases of 47,XXX, 2 cases of 47,XYY and 1 case of 46,X,del(X)(q26.1). Two trisomy 21, two trisomy 18, 2 trisomy 13, and 23 cases of sex chromosomal aneuploidies were verified as false positives. For 9 women with pathological fetal CNVs detected by NIPS, combined fetal karyotyping and CMA has confirmed 1 case of chromosome 13 microdeletion, 1 case of chromosome 18 microduplication, and 1 case of chromosome 18 deletion. For a case with 30 Mb duplication of chromosome 2 and 25 Mb duplication of chromosome 8, CMA analysis had no positive finding, while fetal umbilical cord blood karyotyping has yielded a 46,XX,dup(2)(p23.1p25.3)[13]/46,XX[87] karyotype. The remaining 5 cases were confirmed as false positive results. CONCLUSION: Combined fetal karyotyping and CMA has provided a powerful tool for verifying high-risk fetuses signaled by NIPS.
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Sequenciamento de Nucleotídeos em Larga Escala/métodos , Diagnóstico Pré-Natal , Aneuploidia , Variações do Número de Cópias de DNA , Síndrome de Down , Feminino , Humanos , Cariotipagem , Análise em Microsséries , Gravidez , Síndrome da Trissomia do Cromossomo 13 , Síndrome da Trissomía do Cromossomo 18RESUMO
PURPOSE: The purpose of this study was to describe some prenatal characteristics and laboratory findings of acute fatty liver of pregnancy (AFLP) and provide the clinicians with reasonable predictors and expectation in postpartum recovery. METHODS: At a tertiary referral center 43 patients with AFLP were entered into this retrospective study in 5 years based on the Swansea criteria. Emergent cesarean sections were performed within 24 h, and the criteria of recovery after operation was based on a uniform standard. All of them were hospitalized and treated at the same department of obstetrics and maternal intensive care unit. RESULTS: Prenatally, all women with AFLP had elevated serum hepatic aminotransferase and serum bilirubin levels. Albumin level was decreased in 88 % women and hypoglycemia was documented in 56 % women. Plasma fibrinogen level of 93 % patients was less than 1.75 g/L and prothrombin time (PT) of 91 % was prolonged abnormally. The duration of recovery after delivery ranged from 5 to 20 days. Pearson correlation coefficient between duration of recovery and hyperbilirubinemia was 0.639 (P = 0.001). The levels of PT, plasma fibrinogen and platelet counts were also correlated with the recovery time (R = 0.459, P = 0.002; R = 0.427, P = 0.004; R = 0.435, P = 0.004). Elevated leukocytes, hypoglycaemia, hepatic aminotransferase and uric acid levels showed no value for predicting the prognosis of AFLP (P > 0.01). CONCLUSIONS: AFLP is a rare but serious complication in the third trimester. Prenatal serum bilirubin, PT, plasma fibrinogen levels and platelet counts are the predictors of postpartum recovery, but some Swansea diagnosis criteria do not have the same prognostic significance as others.
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Fígado Gorduroso/sangue , Complicações na Gravidez/sangue , Centros de Atenção Terciária , Adulto , Bilirrubina/sangue , Cesárea , Fígado Gorduroso/terapia , Feminino , Fibrinogênio/análise , Humanos , Unidades de Terapia Intensiva , Contagem de Plaquetas , Período Pós-Parto , Gravidez , Complicações na Gravidez/terapia , Cuidado Pré-Natal , Prognóstico , Tempo de Protrombina , Estudos RetrospectivosRESUMO
BACKGROUND: Because the future application of cell-free fetal DNA screening is expected to dramatically improve the diagnostic yield and reduce unnecessary invasive procedures, it is time to summarize the indications of invasive prenatal diagnosis. This retrospective study was performed to evaluate the changes and efficacies of indications of invasive procedures for detecting cytogenomic abnormalities from 2000 to 2012. MATERIAL AND METHODS: From our regional obstetric unit, 7818 invasive procedures were referred by indications of advance maternal age (AMA), abnormal ultrasound findings (aUS), abnormal maternal serum screening (aMSS), and family history (FH). Chromosome, fluorescence in situ hybridization (FISH), and array comparative genomic hybridization (aCGH) analyses were performed on chorionic villus sampling (CVS) and amniotic fluid (AF) specimens at the Yale Cytogenetics Laboratory. The abnormal findings from single or combined indications were compared to evaluate the diagnostic yield. RESULTS: The annual caseload declined by 57.2% but the diagnostic yield increased from 7.2% to 13.4%. Chromosomal and genomic abnormalities were detected in 752 cases (9.6%, 752/7818) and 12 cases (4%, 12/303), respectively. Significantly decreased AMA referrals and increased aUS and aMSS referrals were noted. The top 3 indications by diagnostic yield were AMA/aUS (51.4% for CVS, 24.2% for AF), aUS (34.7% for CVS, 14.5% for AF), and AMA/aMSS (17.8% for CVS, 9.9% for AF). CONCLUSIONS: Over a period of 13 years, the indication of aMSS and aUS were increasing while AMA was decreasing for prenatal diagnosis of cytogenomic abnormalities, and there was a continuous trend of reduced invasive procedures. Prenatal evaluation using AMA/aUS was the most effective in detecting chromosomal abnormalities, but better indications for genomic abnormalities are needed.
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Aberrações Cromossômicas , Diagnóstico Pré-Natal , Hibridização Genômica Comparativa , Feminino , Humanos , Hibridização in Situ Fluorescente , GravidezRESUMO
The pathogenesis of preeclampsia (PE) has not been elucidated, but immune imbalance is known to be one of the main pathogeneses. Dysfunction of decidual macrophages can lead to PE, and the PD-1/PD-L1 signaling pathway is associated with macrophage polarization. However, the relationship between the influence of the PD-1/PD-L1 signaling pathway on macrophage polarization and the onset of PE has not been fully elucidated. In this study, we analyzed the expression of CD68, iNOS, CD206, PD-1 and PD-L1 and the coexpression of CD68+PD-1+ and CD68+PD-L1+ in the decidual tissue of PE patients (n= 18) and healthy pregnant women (n=20). We found that CD68 and iNOS expression was increased in the decidua of PE patients (P < 0.001) and that CD206, PD-1 and PD-L1 expression and CD68+PD-1+ and CD68+PD-L1+ coexpression were decreased (P < 0.001). To assess the influence of the PD-1/PD-L1 signaling pathway on macrophage polarization, we added an anti-PD-1 mAb (pembrolizumab) or an anti-PD-L1 mAb (durvalumab) during THP-1 differentiation into M1 macrophages. Then, we detected the polarization of CD68+CD80+ macrophages and the expression of iNOS. To examine the effect of macrophage polarization on the invasion ability of trophoblast cells, macrophages were cocultured with HTR8/SVneo cells, and the invasion ability of HTR8/SVneo cells was detected via transwell assays. We found that CD68+CD80+ macrophage polarization was enhanced (P<0.05) and that iNOS expression was greater (P<0.01) in the pembrolizumab group. In the durvalumab group, CD68+CD80+ macrophage polarization and iNOS expression were also increased (P<0.05 and P<0.001). Compared with that in the untreated group, the aggressiveness of HTR8/SVneo cells was decreased in both the pembrolizumab group (P < 0.01) and the durvalumab group (P < 0.001). These findings indicate that the PD-1/PD-L1 signaling pathway may play an important role in the pathogenesis of PE by influencing macrophage polarization and reducing the invasion ability of trophoblasts.
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Antígeno B7-H1 , Decídua , Macrófagos , Pré-Eclâmpsia , Receptor de Morte Celular Programada 1 , Transdução de Sinais , Humanos , Feminino , Pré-Eclâmpsia/imunologia , Pré-Eclâmpsia/patologia , Pré-Eclâmpsia/metabolismo , Gravidez , Antígeno B7-H1/metabolismo , Antígeno B7-H1/imunologia , Receptor de Morte Celular Programada 1/metabolismo , Decídua/imunologia , Decídua/patologia , Decídua/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Transdução de Sinais/imunologia , Adulto , Antígenos CD/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Ativação de Macrófagos/imunologia , Células THP-1RESUMO
The etiology of preeclampsia (PE) is still unknown, and excessive immune activation is an important component of its pathogenesis. Programmed cell death protein 1 (PD-1) is one of immune checkpoints which may prevent overactivated immune attack and lead to a tolerant immune microenvironment. Little is known about the involvement of PD-1-mediated immunoregulation at the maternal-fetal interface in PE. To investigate the inflammatory pattern and the involvement of PD-1 in the decidua of women with PE, decidual tissues were obtained from PE and control pregnant women. Quantitative RT-PCR analysis of the mRNA levels of the inflammatory cytokines was performed. PD-1 expression was detected by immunohistochemistry, western blot analysis, and flow cytometry. To prove the role of PD-1, decidual immune cells were incubated with blocking antibodies, and the inflammatory cytokines were detected by ELISA. We observed that the mRNA levels of IL-1ß, IL-6, TNF-α, and IFN-γ were higher in the decidua of the PE group than in the decidua of the control group. The mRNA levels of IL-4 and IL-10 were lower in PE. The expression level of PD-1 was significantly downregulated, and the proportion (%) of PD-1 + CD45 + cells was significantly lower in PE. There was a significant linear correlation between PD-1 expression and common proinflammatory cytokines in the decidua. Anti-PD-1 blocking antibody significantly increased the secretion of proinflammatory cytokines. Our data suggested that the inflammatory pattern and decreased PD-1 expression in the decidua might play an active role in the local immunoregulatory mechanisms of PE. The PD-1 pathway in the maternal-fetal interface possibly function to break the tolerant immune microenvironment in PE via inflammatory cytokines.
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Citocinas , Pré-Eclâmpsia , Receptor de Morte Celular Programada 1 , Feminino , Humanos , Gravidez , Citocinas/metabolismo , Decídua/metabolismo , Pré-Eclâmpsia/metabolismo , RNA Mensageiro/metabolismo , Receptor de Morte Celular Programada 1/metabolismoRESUMO
BACKGROUND: Preeclampsia (PE) is a common hypertensive pregnancy disorder associated with shallow trophoblast invasion. Although bone morphogenetic protein 2 (BMP2) has been shown to promote trophoblast invasion in vitro, its cellular origin and molecular regulation in placenta, as well as its potential role in PE, has yet to be established. Additionally, whether BMP2 and/or its downstream molecules could serve as potential diagnostic or therapeutic targets for PE has not been explored. METHODS: Placentas and sera from PE and healthy pregnant women were subjected to multi-omics analyses, immunoblots, qPCR, and ELISA assays. Immortalized trophoblast cells, primary cultures of human trophoblasts, and first-trimester villous explants were used for in vitro experiments. Adenovirus expressing sFlt-1 (Ad Flt1)-induced PE rat model was used for in vivo studies. FINDINGS: We find globally decreased H3K27me3 modifications and increased BMP2 signalling in preeclamptic placentas, which is negatively correlated with clinical manifestations. BMP2 is derived from Hofbauer cells and epigenetically regulated by H3K27me3 modification. BMP2 promotes trophoblast invasion and vascular mimicry by upregulating BMP6 via BMPR1A-SMAD2/3-SMAD4 signalling. BMP2 supplementation alleviates high blood pressure and fetal growth restriction phenotypes in Ad Flt1-induced rat PE model. INTERPRETATION: Our findings demonstrate that epigenetically regulated Hofbauer cell-derived BMP2 signalling enhancement in late gestation could serve as a compensatory response for shallow trophoblast invasion in PE, suggesting opportunities for diagnostic marker and therapeutic target applications in PE clinical management. FUNDING: National Key Research and Development Program of China (2022YFC2702400), National Natural Science Foundation of China (82101784, 82171648, 31988101), and Natural Science Foundation of Shandong Province (ZR2020QH051, ZR2020MH039).
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Pré-Eclâmpsia , Trofoblastos , Gravidez , Humanos , Feminino , Ratos , Animais , Trofoblastos/metabolismo , Histonas/metabolismo , Pré-Eclâmpsia/metabolismo , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 2/metabolismo , Proteína Morfogenética Óssea 2/farmacologia , Placenta/metabolismo , Movimento CelularRESUMO
Inadequate trophoblastic infiltration and resulting placental hypoxia and inflammation comprise the core pathological basis of preeclampsia (PE). Maternally expressed gene 3 (MEG3) is known to be involved in the pathogenesis of preeclampsia by inhibiting the migration and invasion of trophoblasts and promoting their apoptosis. Nevertheless, the specific underlying downstream molecular mechanism of MEG3 is less well characterized. In this study, we detected lower expression levels of MEG3 and ß-Catenin and higher expression of nod-like receptor pyrin domain-containing 3 (NLRP3) in placental tissues of pregnant women with severe preeclampsia (sPE) than in normal pregnancies. Elevated serum levels of IL-1ß and TNF-α were also observed in the sPE group. Then, we established a hypoxia/reoxygenation (H/R) model to mimic preeclampsia. Similar results with sPE group were found in the H/R group compared with the control group. In addition, suppressive trophoblast proliferation, migration and invasion and increases in the apoptotic rate and inflammation were also detected in the H/R group. Notably, overexpressing MEG3 markedly improved trophoblast dysfunction and inflammation caused by H/R. However, the effects of MEG3 on trophoblasts, whether upregulated or downregulated, can be reversed by DKK-1 (Wnt/ß-Catenin inhibitor) and MCC950 (NLRP3 inhibitor). The current study revealed that MEG3 regulates trophoblast function and inflammation through the Wnt/ß-Catenin/NLRP3 axis and provided new insights into the pathogenesis of preeclampsia.
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BACKGROUND: Low-grade endometrial stromal sarcoma (LGESS) is a rare indolent tumor with a favorable prognosis. With the importance of improving quality of life recognized, fertility-sparing surgery may be an option for those young women. However, most of the reports suggested that stage IA patients might be candidates for fertility-sparing surgery, and adjuvant hormonal treatment was considered a feasible adjuvant therapy for reducing the recurrence risk of patients with LGESS and hysterectomy was recommended after the completion of pregnancy and delivery. CASE SUMMARY: A 28-year-old pregnant woman diagnosed with stage IB LGESS was treated by fertility-sparing surgery when term cesarean section delivery was performed. Without any adjuvant treatment, she had the other successful term pregnancy and cesarean section 45 mo after first fertility-sparing surgery. Moreover, only hysteroscopic resection was performed to retain fertility again even when the tumor recurred after 6 years. So far the patient's fertility and disease-free status have remained for more than 8 years without any adjuvant therapy despite local resection of the sarcoma. And the two babies were in good health. CONCLUSION: For young patients with stage I LGESS, it seems that repeated fertility-sparing surgeries could be performed even after two term deliveries and the tumor recurrence, and it might be attempted without adjuvant therapy but the counseling should be considered as mandatory.
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OBJECTIVE: To investigate significance and correlation of free fetal DNA (fDNA) and beta-human chorionic gonadotropin (beta-hCG) in circulation in pregnant women with high-risk of Down's syndrome (DS). METHODS: Pregnant women with a male fetus at second trimester screening for Down's syndrome were chosen, including 5 women with a trisomy 21 fetus (DS group), 21 women with DS high-risk pregnant women (DS high-risk group) matched with 22 normal pregnant women as control group. Free fDNA in maternal plasma were extracted. Male DYS14 gene was labled as fDNA, real-time PCR was used to detect fDNA expression. The concentration of beta-hCG in maternal serum was detected by chemiluminescence immune assay. The relationship between level of free fDNA and beta-hCG concentration was analyzed by Pearson correlation analysis. RESULTS: (1) The mean level of free fDNA was (127 +/- 58) GE/ml in DS group, which was significantly higher than (78 +/- 28) GE/ml in DS high-risk group and (48 +/- 21) GE/ml in control group,respectively (P < 0.01). When compared the level of free fDNA between DS high-risk group and control group, it reached statistical difference (P < 0.01). (2) The mean concentration of beta-hCG was (97 +/- 43) kU/L in DS group, which was significantly higher than (58 +/- 25) kU/L in DS high-risk group and (38 +/- 19) kU/L in control group, respectively (P < 0.01). The level of beta-hCG in DS high-risk group was also significantly higher than control group (P < 0.01). (3) The positive relationship between the level of free fDNA in maternal plasma and beta-hCG concentration in maternal serum was observed among three groups (r = 0.83, P < 0.05; r = 0.76,P < 0.01; r = 0.86,P < 0.01). CONCLUSIONS: Free fDNA in maternal plasma might be a candidate marker used for prenatal DS screening. However, its clinical value need to be evaluated because of positive correlation between free fDNA and beta-HCG in maternal circulation.
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Gonadotropina Coriônica Humana Subunidade beta/sangue , DNA/sangue , Síndrome de Down/diagnóstico , Doenças Fetais/diagnóstico , Diagnóstico Pré-Natal/métodos , Adulto , Biomarcadores/sangue , DNA/isolamento & purificação , Síndrome de Down/sangue , Feminino , Doenças Fetais/sangue , Idade Gestacional , Humanos , Masculino , Gravidez , Segundo Trimestre da Gravidez , Gravidez de Alto RiscoRESUMO
The typical hallmark of placenta accreta spectrum (PAS) disorders is increased implantation site intermediate trophoblast (ISIT) cell numbers. However, the extent of trophoblast proliferation and apoptosis have not been found to differ from those of normal placentation. MicroRNA-125a (miR-125a) induces apoptosis in colon cancer cell by targeting myeloid cell leukemia-1 gene (MCL1). We aimed to investigate the influence of miR-125a on ISIT cells in PAS disorders in 15 patients (self-paired trials) with placenta previa and PAS disorders. Expression of miR-125a and MCL1 were measured in villous trophoblasts and basal plate myometrial fibers from creta site and adjacent noncreta tissues by real-time quantitative polymerase chain reaction, and expression of the MCL1 protein was assayed by Western blotting. Flow-cytometry was used to examine the effect of miR-125a overexpression on apoptosis in vitro in HTR-8/SVneo cells, and luciferase activity assays was used to confirm miR-125a targeting of MCL1. In vivo, the expression levels of miR-125a was significantly lower in creta versus noncreta tissues, and the expression of MCL1 was upregulated; moreover, immunohistochemistry showed that the increased ISIT cells in the creta were positive for MCL1 protein. MCL1 was downregulated in the miR-125a-overexpressing HTR-8/SVneo cells in vitro, and overexpression of miR-125a-induced apoptosis in the HTR-8/SVneo trophoblast line. Finally, luciferase activity assays confirmed that miR-125a directly target the 3' untranslated region of MCL1 in the 293T cell line. In conclusion, downregulation of MCL1-targeting miR-125a exerts an antiapoptotic effect on ISIT cells in PAS disorders.
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Apoptose/genética , MicroRNAs/metabolismo , Proteína de Sequência 1 de Leucemia de Células Mieloides/metabolismo , Placenta Acreta/metabolismo , Trofoblastos/metabolismo , Regiões 3' não Traduzidas , Adulto , Linhagem Celular , Proliferação de Células/fisiologia , Regulação para Baixo , Células Epiteliais/metabolismo , Feminino , Humanos , MicroRNAs/genética , Proteína de Sequência 1 de Leucemia de Células Mieloides/genética , Miométrio/metabolismo , Placenta Acreta/genética , GravidezRESUMO
MiR-125b regulates the kinds of cells that undergo apoptosis physiologically and pathologically. However, whether miR-125b affects the apoptotic behavior of trophoblasts and the underlying molecular regulatory mechanisms remains unclear. This study investigated the effect of miR-125b on apoptosis of HTR-8/SVneo cells in vitro. Constructed wild-type reporter vector (Wt-3'UTR) or mutated type reporter vector (Mut-3'UTR) reporter plasmids were transiently transfected into 293â¯T cells along with miR-125b mimics or a negative control. The luciferase reporter assay was used to validate whether the predicted MCL1 gene is a direct target of miR-125b. The HTR8/SVneo cells were transfected with miR-125 mimics, inhibitors, or a scramble control. Real-time polymerase chain reaction and western blotting were used to analyze mRNA and protein expression of the target gene MCL1. Flow cytometry was used to determine the effects on apoptosis. The luciferase activity assay validated the ability of miR-125b to specifically attenuate MCL1 transcription in the 293â¯T cell line, suggesting that MCL1 is a direct target of miR-125b. After transfection by miR-125b, relative expression and translation of the target gene MCL1 mRNA were repressed in HTR-8/SVneo cells. Trophoblast cells were induced to undergo apoptosis by overexpressing miR-125b in the HTR-8/SVneo cell line. In conclusion, MiR-125b may induced apoptosis of HTR8/SVneo cells by targeting MCL1. Our findings suggest that miR-125b may play a pivotal role in the pathophysiology of placentation.
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Apoptose , MicroRNAs/fisiologia , Proteína de Sequência 1 de Leucemia de Células Mieloides/fisiologia , Trofoblastos/fisiologia , Linhagem Celular , HumanosRESUMO
BACKGROUND: Polycystic ovary syndrome (PCOS) is a common endocrine disease characterized by hyperandrogenism, irregular menses, and polycystic ovaries. Several long non-coding RNAs (lncRNAs) are aberrantly expressed in PCOS patients; however, little is known about the effects of the lncRNA-low expression in tumor (lncRNA-LET) on PCOS. We aimed to explore the effects of lncRNA-LET on human granulosa-like tumor cell line, KGN. METHODS: Expression of lncRNA-LET in normal IOSE80 cells and granulosa cells was determined by qRT-PCR. KGN cell viability, apoptosis and migration were measured by trypan blue exclusion method, flow cytometry assay and wound healing assay, respectively. TGF-ß1 was used to induce epithelial-mesenchymal transition (EMT) process. LncRNA-LET expression and mRNA expressions of TIMP2 and EMT-related proteins were measured by qRT-PCR. Western blot analysis was used to measure the protein expression of apoptosis-related proteins, EMT-related proteins, TIMP2, and the proteins in the Wnt/ß-catenin and Notch signaling pathways. RESULTS: lncRNA-LET was down-regulated in KGN cells, and its overexpression inhibited cell viability and migration, and promoted apoptosis in KGN cells. Overexpression of lncRNA-LET increased the expression of E-cadherin and decreased the expressions of N-cadherin and vimentin in KGN cells. These effects of lncRNA-LET on KGN cells were reversed by TIMP2 suppression. Overexpression of TIMP2 inhibited cell viability, migration and EMT process, and increased apoptosis by activating the Wnt/ß-catenin and Notch pathways. CONCLUSION: Overexpression of lncRNA-LET inhibits cell viability, migration and EMT process, and increases apoptosis in KGN cells by up-regulating the expression of TIMP2 and activating the Wnt/ß-catenin and notch signaling pathways.
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Apoptose/genética , Movimento Celular/genética , Transição Epitelial-Mesenquimal/genética , Síndrome do Ovário Policístico/genética , RNA Longo não Codificante/genética , Inibidor Tecidual de Metaloproteinase-2/genética , Linhagem Celular Tumoral , Sobrevivência Celular/genética , Feminino , Tumor de Células da Granulosa/genética , Tumor de Células da Granulosa/patologia , Humanos , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Síndrome do Ovário Policístico/patologia , Regulação para CimaRESUMO
AIM: The purpose of the study was to evaluate the feasibility and reliability of comparative genomic hybridization (CGH) in the detection of genomic imbalances in Chinese malformed fetuses. METHODS: Genomic DNA was extracted from umbilical cord blood or fresh amniotic fluid of 9 malformed fetuses and labeled with SpectrumGreen dUTP or SpectrumRed dUTP. A pair of CGH analyses in which the fluorochromes were exchanged was carried out for each sample. RESULTS: Samples from 9 malformed fetuses were analyzed successfully by CGH. Numerical chromosome aberrations were detected in samples from cases 4, 8 and 9, and were verified by fluorochrome-exchanged CGH. Trisomy 21q was detected in case 4, del 2p24-pter and dup 12p13 was detected in case 8, and del 1p33-pter and del 22q11-12 were detected in case 9. CONCLUSION: CGH is a reliable technique for the detection of genomic imbalances. Fluorochrome-exchanged CGH can reduce inconsistencies in the results caused by deviations in the process of DNA labeling and hybridization, and increase the accuracy and reliability in cases when conventional cytogenetic analysis is unavailable.
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Aneuploidia , Povo Asiático/genética , Anormalidades Congênitas/genética , Feto/anormalidades , Instabilidade Genômica/genética , Hibridização de Ácido Nucleico/métodos , China , Anormalidades Congênitas/etnologia , DNA/genética , Feminino , Corantes Fluorescentes , Humanos , Masculino , Reprodutibilidade dos TestesRESUMO
BACKGROUND: This study aims to assess the diagnostic and prognostic value of Swansea criteria in diagnosing acute fatty liver of pregnancy (AFLP) in a Chinese population. METHODS: A retrospective study was conducted on 52 Chinese women diagnosed with AFLP. All selected cases were reassessed using the Swansea criteria with special focus on the noninvasive criteria, since performing a liver biopsy for this indication is rare in a Chinese population. RESULTS: Ninety point four percent of patients fulfilled five or more of the Swansea criteria. Thirty-one cases were positive for six or more Swansea criteria, but there were no significance differences between patients when using a cutoff criteria <6 or >6. When patients were positive for less than seven criteria, frequency of stillbirth, continuous blood purification (CBP) treatment, hysterectomy, and postpartum hemorrhage were not increased. However, patients who were positive for seven or more criteria had a significantly higher risk of stillbirth and a higher rate of CBP treatment (p < 0.05). Areas under the receiver operating characteristic (ROC) curve of postpartum hemorrhage was 0.670, which reached a statistical significance (p = 0.040). We observed a significantly elevated postpartum hemorrhage along with positivity of the Swansea criteria (p = 0.040). CONCLUSIONS: Swansea criteria without liver biopsy are good screening tools for AFLP diagnosis, and may be useful for assessing disease severity.
Assuntos
Fígado Gorduroso/diagnóstico , Medicina Tradicional Chinesa/métodos , Complicações na Gravidez/diagnóstico , Diagnóstico Pré-Natal/métodos , Adulto , China , Diagnóstico Diferencial , Diagnóstico Precoce , Feminino , Idade Gestacional , Humanos , Testes de Função Hepática/métodos , Gravidez , Estudos Retrospectivos , Sensibilidade e Especificidade , Adulto JovemRESUMO
OBJECTIVE: Placenta accreta is defined as abnormal adhesion of placental villi to the uterine myometrium. Although this condition has become more common as a result of the increasing rate of cesarean sections, the underlying causative mechanism(s) remain elusive. Because microRNA-29a/b/c (miR-29a/b/c) have been shown to play important roles in placental development, this study evaluated the roles of these microRNAs in placenta accreta. METHODS: Expression of miR-29a/b/c and myeloid cell leukemia-1 (MCL1) were quantified in patient tissues and HTR8/SVneo trophoblast cells using the real-time quantitative polymerase chain reaction. Western blotting was used to analyze expression of the MCL1 protein in HTR8/SVneo trophoblast cells with altered expression of miR-29a/b/c. To determine their role in apoptosis, miR-29a/b/c were overexpressed in HTR-8/SVneo cells, and levels of apoptosis were analyzed by flow cytometry. Luciferase activity assays were used to determine whether MCL1 is a target gene of miR-29a/b/c. RESULTS: Expression of miR-29a/b/c was significantly lower in creta sites compared to noncreta sites (p = 0.018, 0.041, and 0.022, respectively), but expression of MCL1 was upregulated in creta sites (p = 0.039). MCL1 expression was significantly downregulated in HTR-8/SVneo cells overexpressing miR-29a/b/c (p = 0.002, 0.008, and 0.013, respectively). Luciferase activity assays revealed that miR-29a/b/c directly target the 3' untranslated region of MCL1 in 293T cells. Over-expression of miR-29a/b/c induced apoptosis in the HTR-8/SVneo trophoblast cell line. Moreover, histopathological evaluation revealed that the number of implantation site intermediate trophoblast (ISIT) cells was increased in creta sites and that these cells were positive for MCL1. CONCLUSIONS: Our results demonstrate that in placenta accreta, miR-29a/b/c inhibits apoptosis of ISIT cells by targeting MCL1. These findings provide new insights into the pathogenesis of placenta accreta.
Assuntos
Apoptose/genética , Regulação para Baixo , Implantação do Embrião/genética , MicroRNAs/metabolismo , Proteína de Sequência 1 de Leucemia de Células Mieloides/metabolismo , Placenta Acreta/metabolismo , Trofoblastos/metabolismo , Linhagem Celular , Vilosidades Coriônicas/metabolismo , Vilosidades Coriônicas/patologia , Feminino , Humanos , MicroRNAs/genética , Proteína de Sequência 1 de Leucemia de Células Mieloides/genética , Miométrio/metabolismo , Miométrio/patologia , Placenta Acreta/genética , Placenta Acreta/patologia , Gravidez , Trofoblastos/patologiaRESUMO
OBJECTIVE: To study the effect and mechanism of the peripheral blood mononuclear cell (PBMC) invasion by HBV on artificial immunization in newborns. METHODS: Fifty-two newborns of HBsAg positive mothers were immunized with HBIG (hepatitis B immunoglobulin) and HBVac (hepatitis B vaccine) and were followed up for 7 months. The newborns' HBV-DNA in serum and in the PBMCs was detected with nested-PCR; anti-HBs was tested with solid phase radioimmunoassay (SP-RIA). PBMCs isolated from newborn peripheral blood were incubated in the presence of PHA or purified HBsAg. Interleukin-2 (IL-2) level in culture supernatants of activated cells was detected by ELISA. RESULTS: The failure rate of immunization was higher in infants with positive HBV-DNA in PBMCs than those with negative HBV-DNA (P < 0.05); IL-2 level in PBMC culture supernatants was lower in former than in the latter and in normal controls (P < 0.05). The level of IL-2 in the immunization failure newborns was lower than that in the successfully immunized newborns and in normal controls (P < 0.05). CONCLUSIONS: Intrauterine invasion of PBMCs by HBV is one of the important reasons for immunization failure in newborns. IL-2 production is closely related to the invasion of PBMCs by HBV, which may contribute to the failure of artificial immunization in newborns.
Assuntos
Vacinas contra Hepatite B/efeitos adversos , Vírus da Hepatite B/fisiologia , Imunização/efeitos adversos , Leucócitos Mononucleares/virologia , DNA Viral/sangue , Antígenos de Superfície da Hepatite B/imunologia , Vírus da Hepatite B/isolamento & purificação , Humanos , Imunoglobulinas/imunologia , Recém-Nascido , Interleucina-2/biossíntese , Interleucina-2/sangueRESUMO
OBJECTIVES: To study the effect and the mechanism of peripheral blood nuclear cells (PBMC) invaded by hepatitis B virus (HBV) on the artificial immunization in newborns. METHODS: Fifty-two newborns, whose mothers were hepatitis B surface antigen (HBsAg) positive, were immunized with hepatitis B immunoglobulin and hepatitis B vaccine (HBVac), and then followed for 7 months. The newborns' serum and PBMC HBV DNA was detected by nested-PCR, hepatitis B surface antibody (HBsAb) was tested with solid phase radioimmunoassay. PBMC from newborn were incubated with PHA and HBsAg. The supernatant interleukin 2 (IL-2) level was measured by enzyme linked immununosorbent assay (ELISA). RESULTS: The rate of vaccination failure was higher in the infants with PBMC HBV DNA positive than those with negative (P < 0.05). The supernatant IL-2 level was lower in the former than that in the latter and the control (P < 0.05). The level of IL-2 in the immunization failure newborns was lower than that in the vaccination success and the control (P < 0.05). CONCLUSIONS: The intrauterine PBMC HBV invasion is one of the important causes of vaccination failure in the newborns. PBMC IL-2 autocrine down regulation is closely related to HBV invasion, that may lead to the failure of HBVac inoculation in the newborns.