Missed Abortion Research Articles

P–389 The relationship between serum hormone profiles and missed abortion in humans

Abstract Study question Are circulating profiles of metabolic-related hormones also associated with the missed abortion (MA) in humans? Summary answer Serum levels of fatty acid-binding protein–4 (FABP4) and fibroblast growth factor 21 (FGF21) are positively associated with MA. What is known already A cluster of endocrine hormones, including FABP4, FGF21, adiponectin, lipocalin–2 (LCN2), exhibit pleiotropic effects on regulating systematic metabolism. Serum levels of them are associated with gestational obesity and diabetes and affect pregnancy outcomes, however, the relationship between their circulating profiles and MA is under-investigated. Study design, size, duration 78 patients with MA and 86 healthy pregnant subjects matching on maternal age and body mass index (BMI) were nested from a prospective cohort in the Chinese population. Participants/materials, setting, methods Fasting serum samples from all participants were collected to test their serum levels of FGF21, FABP4, adiponectin, and LCN2 by enzyme-linked immunosorbent assay method (ELISA). Main results and the role of chance There were no significant differences in circulating profiles of adiponectin and LCN2 between MA patients and healthy pregnant subjects. By contrast, circulating levels of FGF21 and FABP4 were significantly and independently elevated in patients with MA relative to control cases even after adjusting confounding factors (for FGF21: MA: 28.96 ± 2.17 ng/ml; HP: 19.18 ± 1.12 ng/ml, P < 0.001, for FABP4: MA: 152.50 ± 9.31 pg/ml; HP: 90.86 ± 4.14 pg/ml, P < 0.001). Linear regression analysis showed, FGF21 raised every 10 pg/ml contributed to a 24% (95% CI: 15% - 34%) increase in the risk of MA, whereas the OR of FABP4 for the risk of MA was 1.052 (95% CI: 1.022 –1.088). Furthermore, using serum FGF21 level or FABP4 levels discriminated MA from healthy controls with an area under the operating characteristic’s curve (AUROC) of 0.81 (95% CI 0.76–0.92) and 0.70 (95% CI 0.62 - 0.78), respectively. Limitations, reasons for caution The study is limited by the sample size. In addition, our results were based-on Chinese population, whether it could be observed in other ethics group remain to be investigated. Meanwhile, the cause-effect relationship between increased serum FGF21 level and MA remains to be explored. Wider implications of the findings: Our data would suggest that serum levels of FGF21 and FABP4 are associated with MA. Moreover, circulating FGF21 levels may serve as a potential diagnostic biomarker for the recognition of M. Trial registration number IRB Ref. No.: KY201913

Circulating fibroblast growth factor 21 as a potential biomarker for missed abortion in humans

To investigate whether serum levels of fibroblast growth factor 21 (FGF21) and fatty acid-binding protein-4 (FABP4) are associated with missed abortion (MA) in humans. Cross-sectional study. University-affiliated hospital. Patients with MA at 8-12 weeks of gestation. None. Serum levels of FGF21 and FABP4 were tested by enzyme-linked immunosorbent assay. Placental samples were collected during dilation and curettage surgery, and the expression of FGF21 and its related genes were measured using quantitative polymerase chain reaction. In the discovery cohort, 78 patients with MA and 79 healthy pregnant women matched for maternal age and body mass index were nested from a prospective cohort. Circulating levels of FGF21 and FABP4 were significantly and independently elevated in patients with MA relative to the levels in the healthy controls. A single measurement of FGF21 serum level effectively discriminated MA with an area under the receiver operating characteristics curve of 0.80 (95% confidence interval: 0.73-0.87). Importantly, in our external validation cohort that comprised subjects with MA (n = 34) or induced abortion (n = 27), the FGF21 serum levels achieved an area under the receiver operating characteristics curve of 0.85 (95% confidence interval: 0.75-0.96) when identifying those with MA. Nevertheless, expression of FGF21 in the placenta was not associated with its serum concentration. Placental tissues from patients with MA exhibited impaired FGF21 signaling. Our results suggested that serum levels of FGF21 and FABP4 were associated with MA. Circulating FGF21 may serve as a potential biomarker for the recognition of MA.

Could Red Cell Distribution Width and Mean Platelet Volume to Lymphocyte Ratio Predict Missed Abortion?

Missed abortion (MA) is a common pregnancy complication; however, the pathophysiology of MA is still not entirely clear. Mean platelet volume (MPV), and red cell distribution width (RDW) have been found to be associated with obstetrics disorders such as first trimester miscarriage and recurrent miscarriage. We evaluated the association between RDW, MPV to lymphocyte (MPV/L) ratio and MA. A total of 230 cases were included in the present study. Patients were selected from women who had missed abortion (n=125) and controls were selected from the women who had given birth at term (> 37 weeks of gestation) to healthy infants (n=105). In all cases, blood samples were drawn at admission before starting any medication. The MPV, RDW, WBC, L/M ratio and MPV/L ratio levels were significantly higher in the MA group than in the control group. Using multivariate logistic regression analysis, we found that RDW [odds ratio (OR)=1.737, 95% confidence interval (CI)=1.346-2.242, p<0.001] and MPV/L ratio [odds ratio (OR)=1.518, 95% confidence interval (CI) =1.047-2.199, p=0.028] were independent predictors of MA. RDW may be a strong independent predictor of MA.MPV/L ratio was also an independent predictor of MA and it showed superior predictive value than MPV.

Serum Angiopoietin -1 as a biomarker of missed abortion

ABSTRACT Background: Missed abortion (MA) is a pregnancy with an embryo without cardiac pulsation or gestational sac without embryo without attempt for expulsion of the product of conception outside the uterus. Objective: to detect the efficacy of serum Angiopoietin-1(Ang-1) as a biomarker of missed abortion at 6 to 8 weeks of pregnancy Methodology: This case-control study comprised eighty pregnant women aged 20–40 years; they were divided into; A) control group which composed of 40 women with normal viable pregnancy at the 6-8 weeks of pregnancy , B) study group that included 40 women with missed abortion diagnosed by TVUS at same gestational age. Results: This study revealed that, there was highly statistically significant decrease in serum levels of Ang-1 in missed abortion .The mean ±SD of serum Ang-1 were 926.16 ± 211.48 ( 700 – 1620.3) pg/ml in normal pregnant women, and 700.04 ± 87.87 (435.9-797.8) pg/ml in women with missed abortion. The cutoff point was ≤ 780.99 pg/ml with high sensitivity and specificity (87.5.% and 90.00% respectively). There was no correlation between the Ang-1 levels and (age, parity, and body mass index). Conclusion: Serum levels of Ang-1 were highly significantly decreased in a status of missed abortion as early as 6 to 8 weeks of conception when compared with Ang-1 levels in normal viable pregnancies of similar gestational age which indicate its role in pathogenesis of missed abortion. Angiopoietin 1 is not a reliable biomarker for instant diagnosis of missed abortion as some patients need further transvaginal ultrasound (TVUS) and regular follow up.

Distinct mRNA and long non-coding RNA expression profiles of decidual natural killer cells in patients with early missed abortion.

Early non-chromosome-related missed abortion (MA) is commonly associated with an altered immunological environment during pregnancy. Human decidual natural killer (dNK) cells, the most abundant lymphocyte population within the first-trimester maternal-fetal interface, are vital maternal regulators of immune tolerance mediating successful embryo implantation and placentation. Previous studies have shown that dNK cells may play a role in MA. However, the gene expression status and specific altered manifestations of dNK cells in patients with early MA remain largely unknown. Here, we show that MA dNK cells have distinct mRNA and lncRNA expression profiles through RNA sequencing, with a total of 276 mRNAs and 67 lncRNAs being differentially expressed compared with controls. Protein-protein interaction analysis of differentially expressed mRNAs was performed to identify hub genes and key modules. An lncRNA-mRNA regulatory network characterized by the small-world property was constructed to reveal the regulation of mRNA transcription by differential hub lncRNAs. Functional annotation of differentially expressed mRNAs and lncRNAs was performed to disclose their potential roles in MA pathogenesis. Our data highlight several enriched biological processes (immune response, inflammatory response, cell adhesion, and extracellular matrix [ECM] organization) and signaling pathways (cytokine-cytokine receptor interaction, ECM-receptor interaction, Toll-like receptor signaling pathway, and phosphatidylinositol signaling system) that may influence MA. This study is the first to demonstrate the involvement of altered mRNA and lncRNA expression profiles in the dNK cell pathogenesis of early MA, facilitating a better understanding of the underlying molecular mechanisms and the development of novel MA therapeutic strategies targeting key mRNAs and lncRNAs.

Serum Levels of Collectins Are Sustained During Pregnancy: Surfactant Protein D Levels Are Dysregulated Prior to Missed Abortion.

About 15% of pregnant women undergo missed abortion (MA), wherein women do not experience cramping and vaginal bleeding. Dysregulation of the immune molecules and steroid hormones contribute to early pregnancy loss. Collectins- surfactant protein A (SP-A), surfactant protein D (SP-D), and mannose-binding lectin (MBL) are a group of innate immune molecules regulated by the steroid hormones. Reduced levels of SP-A and SP-D during the early gestation exhibited a significant association with the severe early onset preeclampsia. In order to determine the serum profile of collectins throughout the normal pregnancy and explore their predictive potential during the 8-12weeks of gestation for MA, we examined a prospective cohort of pregnant women (n = 221). The serum levels of SP-A and SP-D were significantly downregulated in the normal pregnant women in all the three trimesters (n = 30) compared with the non-pregnant women (n = 20) and were not significantly different across the three trimesters. Fourteen of the women from the cohort underwent MA during the 14-20weeks of gestation and exhibited a significant downregulation in the serum levels of SP-D during 8-12weeks of gestation. A significant inhibition of the HTR-8/SVneo cell proliferation and migration in the presence of a recombinant fragment of human SP-D suggested the relevance of SP-D in placental development. We report here that the serum levels of SP-A, SP-D, and MBL are consistently maintained during pregnancy in the Indian cohort. Dysregulated serum levels of SP-D and P4/E2 ratio during the early first trimester may predict occurrence of MA.

Can neutrophil-to-lymphocyte and monocyte-to-lymphocyte ratios be useful markers for predicting missed abortion in the first trimester of pregnancy?

To explore whether neutrophil-to-lymphocyte ratio (NLR) and monocyte-to-lymphocyte ratio (MLR) could predict missed abortion (MA) at 7-13 week's gestation. A total of 363 women with a diagnosis of MA and 232 women with normal pregnancy at 7-13 week's gestation, who visited our hospital from June 2012 to May 2018 were retrospectively analyzed. At 7 week's gestation, total and differential leukocyte counts, NLR and MLR were compared between women with MA (n = 69) and normal controls (n = 53). The receiver operating characteristic curve was used to select the optimal indicator and its cut-off point. The role of the optimal indicator was further assessed at 8-13 week's gestation. The mean white blood cell counts, the mean neutrophil counts, the median monocyte counts, the mean NLR and the median MLR in women with MA were significantly lower than those in normal controls (P < 0.05, respectively).The neutrophil counts had the highest area under the curve (AUC) value of 0.772 (95% confidence interval 0.675-0.869) with a cut-off value of 4.870 × 109 /L, and the sensitivity was 72.46%, the specificity was 69.81%, positive predictive value was 75.76%, and negative predictive value was 66.07%. In addition, the neutrophil counts were also significantly lower in MA groups than those in normal controls at 8-13 week's gestation, and all had the highest AUC values. Neutrophil counts may predict MA in the first trimester of pregnancy, which may provide a promising marker to diagnose missed abortion as early as 7 week's gestation.

Excessive reactive oxygen species induce apoptosis via the APPL1-Nrf2/HO-1 antioxidant signalling pathway in trophoblasts with missed abortion

AimsPrevious evidence has demonstrated that oxidative stress is related to the pathogenesis of missed abortion (MA), but the specific mechanism remains obscure. The adaptor protein APPL1 is one of the differential proteins in chorionic trophoblasts. Thus, this study aimed to assess the potential influence of APPL1 on oxidative stress responses as well the possible molecular mechanisms involving in MA. Main methodsIn the present study, the chorionic trophoblasts and the HTR-8/SVneo cell line were researched in vitro. Small interfering RNA (siRNA) was used to suppress the expression of APPL1. The fluorescent probes DHE and DCFH-DA were used to assess the intracellular reactive oxidative species (ROS). The activity of superoxide dismutase (SOD) was determined. Apoptosis was detected by TUNEL and flow cytometry. Cell viability was determined using Cell Counting Kit-8. Protein expression was detected by immunohistochemistry, western blotting, and reverse transcription-quantitative PCR. Key findingsThe application of oxidant in normal chorionic trophoblasts induced cell death and overproduction of ROS, which was consistent with MA. In addition, knockdown of APPL1 in HTR-8/SVneo cells resulted in increased ROS and apoptosis, which could be rescued by pretreatment with antioxidants. Mechanistically, we report that overproduction of ROS in trophoblasts and disturbed SOD, APPL1 and Nrf2/HO-1 antioxidant responses constitute important contributors to apoptosis. SignificanceOur results suggest that APPL1 has antioxidant properties that suppress oxidative stress and apoptosis via the Nrf2/HO-1 pathway. Moreover, antioxidant N-acetylcysteine (NAC) effectively restored the impaired antioxidative defense system elicited by excess ROS, as a potential therapeutic reagent for MA.

MiR-98 is involved in missed abortion by targeting GDF6 and FAPP2.

Missed abortion (MA) is a common disease in obstetrics and gynecology. More and more studies have focused on the relationship between miRNAs and pregnancy maintenance and its related diseases. The aim of this article is to explore the relationship between miRNA and MA. The expression of miR-98 were detected by in situ hybridization and real-time PCR. Cell proliferation, activity and migration were measured via Edu, MTT, and transwell assays. The target genes of miR-98 are identified by dual-luciferase activity assay. And the expression levels of target genes were determined by Western blot, real-time PCR and immunohistochemistry. miR-98 was significantly up-regulated in placental villi from over 35 years old MA patients compared with the age-matched normal pregnant women. Up-regulation of miR-98 suppressed the proliferation, activity and migration of the human trophoblast HTR-8/SVneo cell in vitro. miR-98 could bind to GDF6 and FAPP2 mRNA 3'-UTR and negatively regulate their expression. The downregulation of miR-98 promoted cell proliferation, then knockdown of GDF6 or FAPP2 inhibited miR-98-mediated cell proliferation. GDF6 and FAPP2 expression in the placental villi from MA patients were decreased compared to normal placental tissues. The expression of miR-98 in MA had an opposite relationship with the expression of GDF6 and FAPP2. Overexpression of miR-98 is associated with the occurrence of MA. miR-98 prevents proliferation, viability and migration of trophoblast cells partially through targeting GDF6 and FAPP2.

Pigment epithelium‑derived factor facilitates NLRP3 inflammasome activation through downregulating cytidine monophosphate kinase 2: A potential treatment strategy for missed abortion.

A number of conditions may underlie the occurrence of missed abortion (MA), including inflammation. Pigment epithelium-derived factor (PEDF) is a novel mediator of the inflammation-related nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome, which is associated with several human diseases. However, the association between MA and NLRP3 inflammasome, and whether PEDF is reduced in MA, remain unknown. In the present study, the decidua and chorion tissues of patients who had suffered a MA were examined, and a lipopolysaccharide (LPS)-induced human chorionic trophoblast HTR8/SVneo cell model was established to mimic MA in vitro. The results revealed that cytidine monophosphate kinase 2 (CMPK2) expression and NLRP3 inflammasome activation, downstream pro-IL-18 and pro-IL-1β expression, and IL-18 and IL-1β release, were all significantly increased in MA tissues or LPS-induced HTR8/SVneo cells. PEDF reversed the increase in CMPK2 expression and activation of the NLRP3 inflammasome axis and, thus, downregulated the production of mitochondrial reactive oxygen species and mitochondrial DNA release, resulting in reduced lactate dehydrogenase release, and a resultant decrease in cell viability. Recovery of CMPK2 expression abolished all the effects of PEDF, indicating that CMPK2 may be an effector downstream of PEDF. PEDF reduced CMPK2 protein levels but did not affect the mRNA levels, and treatment with the proteasomal inhibitor MG132 significantly reversed this reduction in CMPK2 protein levels. Furthermore, a ubiquitination assay of immunoprecipitation demonstrated that CMPK2 was polyubiquitinated in the presence of LPS, PEDF and MG132. These results indicated that the NLRP3 inflammasome is implicated in the pathogenesis of MA, and PEDF may reduce MA through ubiquitin-dependent proteasomal degradation of CMPK2 to inhibit NLRP3 activation, which may serve as a novel strategy for preventing or reducing the risk of MA.

Evaluation of Angiopoietin One and Angiopoietin Two with Missed Abortion

Missed abortion (MA) is a type of miscarriage, refer to pregnancy in which there is fetal demise without intervention, and also uterine that may expel the product of conception prior to 20 weeks of gestation to assess the role of ANG1 and ANG2 level in early pregnancy and compare these level with healthy pregnancy women, ANG1 play role in new blood vessels maturation and stabilization, the inhibition of endothelial apoptosis and reduction of vascular permeability in stable environment. ANG2 is an antagonist of ANG1 and it known to enhance the plasticity, destabilization and permeability of blood vessels and vascular remodeling site. A prospective cross - study, all women were attended to obstetrics and gynecology outpatient,and this study was carried out from Jan 2019 to July 2019. Sixty Women with missed miscarriage fetus heart negative 6-8 weeks were screened to participate in the present study group, Ages were between 18 – 35 years old, and they were from center and the periphery of Kirkuk city to comparative forty apparently health women with early pregnancy fetus heart positive a control groups. A serum level of ANG1 and ANG2 was measured by ELIZA. The result shows that the mean serum level of Angiopoietin one was significantly decrease in missed mischarge compared to control women (2.92±2.4 vs. 5.6±3.2 ng/ml) respectively at a P &lt; 0.05. And this study showed that there was the significant decrease serum level of angiopoietin two in missed miscarriage patients compared with the control group women (8.12±3.5 vs. 16.81 ±12.3). This study showed that there were the positive correlations of Angiopoietin one and two with missed miscarriage. It can be conclude that the level of serum Angiopoietin -1 and level of serum angiopoietin -2 in missed abortion decrease and it is can be used as an early and effective biomarker for diagnosis of missed abortion.

Comprehensive analysis of the differential expression profile of microRNAs in missed abortion.

To screen the key circulating microRNAs (miRNAs) involved in missed abortion (MA) and explore their role in MA process. We examined the miRNA profile from the serum of three MA patients and three early pregnancy induced abortion patients (controls) by next-generation sequencing. We analyzed the target genes of the differentially expressed (DE) miRNAs to analyze the function and pathway enrichment using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes, respectively. We validated five candidate miRNAs by real time-qPCR. Integrated miRNA-mRNA-pathway network analysis was performed to show the interaction network of the candidate miRNAs and their target genes of interest with the involved pathways. It was observed that 227 miRNAs were differently expressed between the MA group and the early pregnancy control group, with 58 miRNAs downregulated and 169 miRNAs upregulated in the MA group. Real-time qPCR results revealed that expression of the five candidate miRNAs, namely hsa-miR-22-3p, hsa-miR-145-3p, hsa-miR-107, hsa-miR-361-3p, and hsa-miR-378c, was consistent with the miRNA data obtained by sequencing. Integrated miRNA-mRNA-pathway network analysis illustrated that target genes of the candidate miRNAs were mainly involved in the PI3K-Akt signaling pathway, HIF-1 signaling pathway, and VEGF signaling pathway, which would have potential significance in pregnancy and MA. We are the first to reveal the DE miRNAs involved in MA and illustrate their functional interaction network. These results might provide potential circulating biomarkers and new therapeutic targets for MA.

Serum and endometrial organic acid profile in women with missed abortion

Aim. To determine the role of organic acids in the pathogenesis of missing abortion.Materials and Methods. To analyze the endometrial organic acid content, we applied high performance liquid chromatography. Sampling was performed from 20 women with missed abortion and 10 patients with normal pregnancy utilising the vacuum aspiration. Following protein separation by means of double centrifugation and ultrafiltration, supernatant was profiled for the organic acids.Results. Women who experienced missing abortion had significantly higher concentrations of serum and endometrial lactic acid concentrations (10.01 ± 0.16 and 8.28 ± 0.20 mmol/L, respectively) than those with healthy pregnancy (8.16 ± 0.24 and 7.57 ± 0.20 mmol/L, respectively). In contrast, endometrial pyruvic acid level was lower in abortion group (27.71 ± 0.04 mmol/L) as compared to the controls (34.1 ± 0.40 mmol/L). Furthermore, 4-hydroxyphenylacetic acid was found exclusively in women with missed abortion.Conclusion. Changes in organic acid profile may play a major role in pathogenesis of missed abortion. Serum and endometrial 4-hydroxyphenylacetic acid can be suggested as a novel biomarker of early pregnancy loss.

Expression of galectin-3 and apoptosis in placental villi from patients with missed abortion during early pregnancy.

The success of a pregnancy relies on moderate trophoblast apoptosis. If the ‘inhibition-induction’ balance of apoptosis is broken, a pathological pregnancy may occur. Galectin-3 has an important role in numerous biological processes, including tumor cell apoptosis. However, the association between galectin-3 and missed abortion (MA) has remained elusive. In the present study, the mRNA and protein expression levels of galectin-3 in placental villi, and the apoptotic index of placental cells from patients with MA were assessed and compared with those in a normal spontaneous abortion group. The present study investigated the function of galectin-3 in the process of MA and the possible association between placental apoptosis and galectin-3 expression in MA patients. Furthermore, the role of galetin-3 in patients with MA at different times (<4 and >4 weeks) was explored. The present study provided a potential mechanism of MA from a perspective of apoptosis and also provided potential therapeutic approaches to prevent MA.

Whole‑exome sequencing analysis of products of conception identifies novel mutations associated with missed abortion.

Missed abortion (MA) refers to a pregnancy in which there is fetal demise without outside intervention, and additionally no uterine activity that may expel the product of conception (POC) prior to 20 weeks of gestation. Chromosomal abnormalities are the primary cause of MA and single gene defects in the POC may additionally be associated with MA; however, few studies have been conducted on the identification of mutations by whole-exome sequencing. In the present study, 19 unrelated MA POCs were collected and whole-exome sequencing was performed on the POC. Bioinformatics analysis was performed on sequence variants from a list of 286 selected candidate genes that were associated with early embryonic lethality and MA. A total of 36 sequence variants in 32 genes potentially associated with MA were identified in 15 out of 19 patients. Gene Ontology analysis suggested that these genes were enriched in biological processes in early embryonic development, including ‘chordate embryonic development’, ‘cell proliferation’ and ‘forebrain development’. Further strict in silico bioinformatics analysis predicted that the LIM domain-binding protein 1 (c.662C>T; p.S221L) variant was a highly pathogenic variant. In conclusion, the results of the present study provide researchers and clinicians with a better understanding of the etiology and molecular mechanism of human embryonic lethality and MA.

Evaluation of thiol/disulphide homeostasis as a novel predictor testing tool of early pregnancy viability

ObjectiveTo evaluate serum dynamic thiol/disulphide concentrations in patients with suspected missed abortion (MA) and to determine whether this ratio has a predictive role in the viability in these pregnancies. Materials and MethodsIn this prospective cohort study, 48 out of 120 recruited pregnant patients were confirmed MA in the study group. Following the recommended waiting time (7–10 days), the remaining 72 viable pregnancies that met the inclusion criteria were categorized as the control group. A novel, automated, and spectrophotometric assay, which can measure both sides of the thiol/disulphide balance, was used. The cut-off values were used for the ROC curve. ResultsThere were no statistically significant differences between the groups (MA and control) regarding maternal obstetric and demographic features. Significantly reduced levels of Total Thiol and Native Thiol were shown in patients with MA compared to the control group (p = 0.016 and p = 0.001, respectively). Serum levels of disulphide was significantly increased in the MA group (23.4 ± 7.8 μmol/l vs. 17.9 ± 4.99 μmol/l, p < 0.0001). Disulphide values of less than 17.68 predicted 80.8% of the viable pregnancies. ConclusionSignificantly increased serum disulphide levels, one of the oxidative stress markers, and decreased antioxidant levels (total and native thiol) were found in patients with MA. Increased oxidative stress status is thought to play a role in the etiology of MA. Serum dynamic thiol/disulphide homeostasis may serve as a promising testing tool to rule out subsequent diagnosis of MA and may benefit as an early pre-treatment testing tool for viability.

Granulysin-mediated apoptosis of trophoblasts in blighted ovum and missed abortion.

Granulysin (GNLY) is a cytotoxic molecule mostly present in decidual natural killer (NK) cells. Blighted ovum (BO) and missed abortion (MA) represent the early pathological pregnancies with hindered development of the embryoblast or a dead embryo. We investigated the GNLY-mediated apoptotic mechanism potentially responsible for delayed termination of pregnancy. We performed immunohistological and immunofluorescence labeling of decidual tissues (GNLY, Apaf-1, NF-κB). NKG2A expression was analyzed by flow cytometry and GNLY mRNA by RT-qPCR. The GNLY labeling intensity (H score) was lower in the nuclei of trophoblast cells in BO and MA. GNLY gene levels were inversely detected in BO and MA. A decreased decidual NK cell percentage was found in MA. NK cells from pathological pregnancies expressed lower NKG2A levels. The highest frequency of Apaf-1 was found in trophoblast cells of MA. NF-kB was highly expressed in decidual cells of BO. The reduced activation of GNLY-mediated killing might be implicated in the slower rejection of trophoblast cells in BO and MA. A decreased authentic decidual NK cell number could be responsible for low cytotoxicity against trophoblast cells in MA. In BO, trophoblast cells have a higher survival potential due to increased NF-kB expression.

NMR-based serum metabolomics study reveals a innovative diagnostic model for missed abortion

A missed abortion (MA) is an in-utero death of the embryo or fetus before the 20th week of gestation with retained products of conception. In order to discover novel biomarkers for MA, a 1H NMR spectroscopy-based metabolomics approach was applied to detect human MA serum metabolic profiles. Serum samples were obtained from patients with MA (n = 15) and healthy controls (n = 9) for study. The NOESYPR1D spectrum combined with multi-variate pattern recognition analysis was used to cluster the groups and establish a disease-specific metabolites phenotype. Principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) models were capable of distinguishing MA patients from healthy subjects. The results revealed that 24 metabolites altered in MA patients compared with the control population. Metabolomic pathway analysis demonstrated that alanine, aspartate and glutamate metabolism, citrate cycle (TCA cycle), taurine and hypotaurine metabolism were significantly altered in MA. The results indicated that serum NMR-based metabolomic profiling method is sensitive and specific enough to distinguish MA and from healthy controls, this method could be developed as a clinically useful diagnostic tool for MA. The finding from the MA serum metabolic profiling shed a new light on further understanding of MA disease mechanisms.

Abnormal expression of microRNA‑575 leads to missed abortion through regulating apoptosis and angiogenesis

Numerous microRNA (miR) are important for placental development and function. miR-575 has been demonstrated to be upregulated in maternal placenta in patients who have experienced a miscarriage. The present study aimed to explore the role of abnormal expression of miR-575 in missed abortion (MA) and to further analyze the potential molecular mechanisms. Embryo villus tissue samples were extracted from 10 childless women with MA and 10 fertile women without a history of MA. Additionally, human choriocarcinoma cells, JEG-3, were used in the present study, which were transfected with miR-575 mimic, inhibitor and scramble. The expression of miR-575 in embryo villus tissues and in JEG-3 cells was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Apoptosis in villus tissues of patients with MA and in JEG-3 cells of miR-575 mimic, inhibitor and scramble groups were detected by flow cytometry. Furthermore, the expression levels of apoptosis-related proteins, including B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) and phosphorylated (p)-p53, and angiogenesis-related proteins, including vascular endothelial growth factor (VEGF) and angiopoietin 2 (Ang-2), were measured by RT-qPCR and western blotting. Additionally, the target of miR-575 was predicted and clarified by luciferase reporter assay. miR-575 was significantly overexpressed in MA villus tissue compared with normal tissue (P<0.05). The percentage of apoptotic cells in MA embryo villus tissue was significantly higher than that in normal tissue (P<0.05). After JEG-3 cells were transfected with miR-575 inhibitor, the expression of miR-575 and the percentage of apoptotic cells decreased significantly compared with the control (P<0.05). MiR-575 suppression significantly increased the expression of Bcl-2 (P<0.05), and decreased the expressions of Bax (P<0.05) and p-p53 (P<0.01) compared with the control. Furthermore, miR-575 suppression significantly increased the expressions of angiogenesis-related proteins, Ang-2 and VEGF (P<0.01). Superoxide dismutase 2 was identified as the target of miR-575. Therefore, abnormal expression of miR-575 may lead to MA through regulating apoptosis and angiogenesis. Inhibition of miR-575 may inhibit apoptosis and promote angiogenesis in MA.

Lidocaine and Dexamethasone for Paracervical Block Anesthesia in Women with Missed Abortion : Randomized Controlled Trial

Aim of the work: nerve block is a technique whereby local anesthetic solutions are infiltrated around a nerve (or perineurally) to provide anesthesia and analgesia. Nerve block for intraoperative and postoperative pain management is associated with improved analgesia, fewer opioid-related adverse events, earlier ambulation and shorter hospital stay when compared to intravenous opioid analgesia alone.This study aimed to assess the efficacy of adding dexamethasone to lidocaine for cervical block anesthesia for prolonging the duration and anesthetic effect in women with missed abortion undergoing vacuum evacuation. Patients and methods: this study is a randomized controlled trial and it was conducted in accordance with the ethical committee protocols and informed consent procedures of Ain Shams University Maternity Hospital during the period between Augusts to December 2016.Sample size was calculated using PASS® version 11 programs, setting the type-1 error (α) at 0.05 and the power (1-β) at 0.8. Conclusion: para cervical block can be used as a safe and effective anesthetic technique in patients who need surgical uterine evacuation of missed abortion. Adding dexamethasone can increase effectiveness and duration of para cervical block. Intraoperative pain level was accepted in 80% of patients, these patients had no or mild to moderate accepted pain. We did not detect any postoperative complications in our patients including (excessive vaginal bleeding, hematoma or general manifestations of lidocaine toxicity) and It is recommended to apply PCB for cases of first trimester missed abortion who require uterine suction evacuation. Lidocaine is preferably mixed with dexamethasone to have better results as regards pain score. It is the anesthetic method of choice especially when general anesthesia is a high risk procedure.