GPR124 promotes trophoblast proliferation, migration, and invasion and inhibits trophoblast cell apoptosis and inflammation via JNK and P38 MAPK pathways.

Early-onset preeclampsia, which occurrs before 34 weeks of gestation, is the most dangerous classification of preeclampsia, which is a pregnancy-specific disease that causes 1% of maternal deaths. G protein-coupled receptor 124 (GPR124) is significantly expressed at various stages of the human reproductive process, particularly during embryogenesis and angiogenesis. Our prior investigation demonstrated a notable decrease in GPR124 expression in the placentas of patients with early-onset preeclampsia compared to that in normal pregnancy placentas. However, there is a lack of extensive investigation into the molecular processes that contribute to the role of GPR124 in placenta development. This study aimed to examine the mechanisms by which GPR124 affects the occurrence of early-onset preeclampsia and its function in trophoblast. Proliferative, invasive, migratory, apoptotic, and inflammatory processes were identified in GPR124 knockdown, GPR124 overexpression, and normal HTR8/SVneo cells. The mechanism of GPR124-mediated cell function in GPR124 knockdown HTR8/SVneo cells was examined using inhibitors of the JNK or P38 MAPK pathway. Downregulation of GPR124 was found to significantly inhibit proliferation, invasion and migration, and promote apoptosis of HTR8/SVneo cells when compared to the control and GPR124 overexpression groups. This observation is consistent with the pathological characteristics of preeclampsia. In addition, GPR124 overexpression inhibits the secretion of pro-inflammatory cytokines interleukin (IL)-8 and interferon-γ (IFN-γ) while enhancing the secretion of the anti-inflammatory cytokine interleukin (IL)-4. Furthermore, GPR124 suppresses the activation of P-JNK and P-P38 within the JNK/P38 MAPK pathway. The invasion, apoptosis, and inflammation mediated by GPR124 were partially restored by suppressing the JNK and P38 MAPK pathways in HTR8/SVneo cells. GPR124 plays a crucial role in regulating trophoblast proliferation, invasion, migration, apoptosis, and inflammation via the JNK and P38 MAPK pathways. Furthermore, the effect of GPR124 on trophoblast suggests its involvement in the pathogenesis of early-onset preeclampsia.

Down-regulated Wnt7a and GPR124 in early-onset preeclampsia placentas reduce invasion and migration of trophoblast cells.

OBJECTIVES: Preeclampsia (PE) is a disease specific to pregnancy that causes 9-10 % of maternal deaths. Early-onset PE (<34 weeks' gestation) is the most dangerous category of PE. Wnt7a and GPR124 (G protein-coupled receptor 124) are widely expressed in the human reproductive process. Especially during embryogenesis and tumorigenesis, Wnt7a plays a crucial role. However, few studies have examined the association between Wnt7a-GPR124 and early-onset PE. The aim of this study was to examine the significance of Wnt7a and GPR124 in early-onset PE as well as Wnt7a's role in trophoblast cells. METHODS: Immunohistochemistry (IHC), real-time PCR, and western blotting (WB) were used to investigate Wnt7a and GPR124 expression in normal and early-onset PE placentas. Additionally, FACS, Transwell, and CCK-8 assays were used to diagnose Wnt7a involvement in migration, invasion, and proliferation. RESULTS: In the early-onset PE group, Wnt7a and GPR124 expression was significantly lower than in the normal group, especially in the area of syncytiotrophoblasts (STBs) and extravillous trophoblasts (EVTs). A negative correlation was found between Wnt7a RNA and GPR124 expression (r=-0.42, p<0.01). However, the Wnt7a RNA expression level was positive correlated with PE severity. In further cellular functional experiments, knockdown of Wnt7a inhibits HTR8/SVeno cells invasion and migration but has little effect on proliferation and apoptosis. CONCLUSIONS: Through the Wnt pathway, Wnt7a regulates trophoblast cell invasion and migration, and may contribute to early-onset preeclampsia pathogenesis. A molecular level study of Wnt7a will be needed to find downstream proteins and mechanisms of interaction.

Mediation effect of maternal triglyceride and fasting glucose level on the relationship between maternal overweight/ obesity and fetal growth: a prospective cohort study.

BACKGROUND: Previous studies have suggested that maternal overweight/obesity is asscociated with macrosomia. The present study aimed to investigate the mediation effects of fasting plasma glucose (FPG) and maternal triglyceride (mTG) in the relationship between maternal overweight/obesity and large for gestational age (LGA) among non-diabetes pregnant women. METHODS: This prospective cohort study was conducted in Shenzhen from 2017 to 2021. A total of 19,104 singleton term non-diabetic pregnancies were enrolled form a birth cohort study. FPG and mTG were measured at 24-28 weeks. We analyzed the association of maternal prepregancy overweight/obesity with LGA and mediation effects of FPG and mTG. Multivariable logistic regression analysis and serial multiple mediation analysis were performed. The odds ratio (OR) and 95% confidence intervals (CIs) were calculated. RESULTS: Mothers who were overweight or obese had higher odds of giving birth to LGA after adjusting potential confounders (OR:1.88, 95%CI: 1.60-2.21; OR:2.72, 95%CI: 1.93-3.84, respectively). The serial multiple mediation analysis found prepregnancy overweight can not only have a direct positive effect on LGA (effect = 0.043, 95% CI: 0.028-0.058), but also have an indirect effect on the LGA through two paths: the independent mediating role of FPG (effect = 0.004, 95% CI: 0.002-0.005); the independent mediating role of mTG (effect = 0.003,95% CI: 0.002-0.005). The chain mediating role of FPG and mTG has no indirect effect. The estimated proportions mediated by FPG and mTG were 7.8% and 5.9%. Besides, the prepregnancy obesity also has a direct effect on LGA (effect = 0.076; 95%CI: 0.037-0.118) and an indirect effect on LGA through three paths: the independent mediating role of FPG (effect = 0.006; 95%CI: 0.004-0.009); the independent mediating role of mTG (effect = 0.006; 95%CI: 0.003-0.008), and the chain mediating role of FPG and mTG (effect = 0.001; 95%CI: 0.000-0.001). The estimated proportions were 6.7%, 6.7%, and 1.1%, respectively. CONCLUSION: This study found that in nondiabetic women, maternal overweight/obesity was associated with the occurence of LGA, and this positive association was partly mediated by FPG and mTG, suggesting that FPG and mTG in overweight/obese nondiabetic mothers deserve the attention of clinicians.

Maternal BMI changes from the prepregnancy to postpartum period are associated with postpartum cardiometabolic risk factors: a longitudinal study.

PURPOSE: This study aimed at investigating the associations between the total body mass index (BMI) change at 3 or 4 years postpartum compared to the prepregnancy and cardiometabolic risk factors. METHODS: This longitudinal study included 1305 participants. Based on the total postpartum BMI changes, they were divided into < 0 units, 0-1.7 units, and > 1.7 units groups using the interquartile range. Multiple linear regression models were used to analyze the associations. RESULTS: Compared to the reference group, there was a progressive increase in the ßcoefficient (ßcoef) of homeostasis model assessment of insulin resistance (HOMA-IR) of cardiometabolic risk in the following groups: the '0-1.7 units' group with the 'overweight traj' [ßcoef 0.33; 95% confidence intervals (CI) 0.22, 0.44)] or the 'obesity traj' [0.66; (0.45, 0.88)] and the '> 1.7 units' group with the 'normal traj' [0.33; (0.22, 0.44)], the 'overweight traj' [0.54; (0.41, 0.67)] or the 'obesity traj' [0.97; (0.79, 1.15)]. The same increasing trend of ßcoef was also found in DBP, FPG, LDL, WHR, BF%. However, the '< 0 units' group with the 'low traj' [0.13; (0.06, 0.21)] and the '0-1.7 units' group with the 'low traj' [0.08; (0.03, 0.13)] had higher high-density lipoprotein cholesterol (HDL-C) level than the reference group. CONCLUSION: Women with a postpartum BMI gain > 1.7 units are positively associated with cardiometabolic risk factors, especially for those in the 'obesity traj' or 'traj D'. Conversely, women with a postpartum BMI loss > 0 units have negative association with cardiometabolic risk factors, especially for those in the 'low traj' or 'traj B'.

Elevated trophoblastic Siglec6 contributes to the impairment of vascular endothelial cell functions by downregulating Wnt6/ß-catenin signaling in preeclampsia.

Preeclampsia (PE), a systemic vascular disorder, is the leading cause of maternal and perinatal morbidity and mortality, and its pathogenesis has yet to be fully elucidated. Siglec6, a transmembrane protein, is highly expressed in human placental trophoblasts, and previous studies have shown that Siglec6 overexpression correlates with PE, but the role of Siglec6 during PE progression is unknown. Here, we demonstrated that the mRNA and protein expression levels of Siglec6 were upregulated in early-onset PE placentas compared with uncomplicated pregnancies, and Siglec6 was primarily located in syncytiotrophoblasts (STBs) and extravillous trophoblasts (EVTs). Moreover, our results showed that chemical reagent-induced HIF-1α accumulation promoted the mRNA and protein levels of Siglec6 in HTR8/SVneo and BeWo cells. Although Siglec6 overexpression did not affect HTR8/SVneo cell proliferation, migration, and invasion, the conditional medium derived from the Siglec6 overexpressed HTR8/SVneo cells (Siglec6-OE-CM) significantly impaired the proliferation, migration, invasion, and tube formation of human umbilical vein endothelial cells (HUVECs). Subsequently, the transcriptome sequencing results revealed that Siglec6 overexpression led to the downregulation of Wnt6 in HTR8/SVneo cells, which was further confirmed by qPCR and ELISA. Recombinant human Wnt6 reversed Siglec6-OE-CM-mediated suppression of HUVEC functions by reactivating the Wnt/ß-catenin signaling pathway. Altogether, our study found that elevated trophoblastic Siglec6 contributed to the impairment of vascular endothelial cell functions by downregulating Wnt6/ß-catenin signaling.

Placental trophoblast-specific overexpression of chemerin induces preeclampsia-like symptoms.

Maternal circulating levels of the adipokine chemerin are elevated in preeclampsia, but its origin and contribution to preeclampsia remain unknown. We therefore studied (1) placental chemerin expression and release in human pregnancy, and (2) the consequences of chemerin overexpression via lentivirus-mediated trophoblast-specific gene manipulation in both mice and immortalized human trophoblasts. Placental chemerin expression and release were increased in women with preeclampsia, and their circulating chemerin levels correlated positively with the soluble Fms-like tyrosine kinase-1 (sFlt-1)/placental growth factor (PlGF) ratio, a well-known biomarker of preeclampsia severity. Placental trophoblast chemerin overexpression in mice induced a preeclampsia-like syndrome, involving hypertension, proteinuria, and endotheliosis, combined with diminished trophoblast invasion, a disorganized labyrinth layer, and up-regulation of sFlt-1 and the inflammation markers nuclear factor-κB (NFκB), tumor necrosis factor (TNF)-α, and interleukin (IL)-1ß. It also led to embryo resorption, while maternal serum chemerin levels correlated negatively with fetal weight in mice. Chemerin overexpression in human trophoblasts up-regulated sFlt-1, reduced vascular endothelial factor-A, and inhibited migration and invasion, as well as tube formation during co-culture with human umbilical vein endothelial cells (HUVECs). The chemokine-like receptor 1 (CMKLR1) antagonist α-NETA prevented the latter phenomenon, although it did not reverse the chemerin-induced down-regulation of the phosphoinositide 3-kinase/Akt pathway. In conclusion, up-regulation of placental chemerin synthesis disturbs normal placental development via its CMKLR1 receptor, thereby contributing to fetal growth restriction/resorption and the development of preeclampsia. Chemerin might be a novel biomarker of preeclampsia, and inhibition of the chemerin/CMKLR1 pathway is a promising novel therapeutic strategy to treat preeclampsia.

Association of maternal obesity with preterm birth phenotype and mediation effects of gestational diabetes mellitus and preeclampsia: a prospective cohort study.

BACKGROUND: The association between maternal obesity and preterm birth remains controversial and inconclusive, and the effects of gestational diabetes mellitus (GDM) and preeclampsia (PE) on the relationship between obesity and preterm birth have not been studied. We aimed to clarify the relationship between prepregnancy body mass index (BMI) and the phenotypes of preterm birth and evaluate the mediation effects of GDM and PE on the relationship between prepregnancy BMI and preterm birth. METHODS: We conducted a prospective cohort study of 43,056 women with live singleton births from 2017 through 2019. According to the WHO International Classification, BMI was classified as underweight (BMI < 18.5 kg/m2), normal weight (BMI 18.5-25 kg/m2), overweight (BMI 25-30 kg/m2) and obese (BMI ≥30 kg/m2). Preterm birth was defined as gestational age less than 37 weeks (extremely, < 28 weeks; very, 28-31 weeks; and moderately, 32-36 weeks). The clinical phenotypes of preterm birth included spontaneous preterm birth (spontaneous preterm labor and premature rupture of the membranes) and medically indicated preterm birth. We further analyzed preterm births with GDM or PE. Multivariable logistic regression analysis and causal mediation analysis were performed. RESULTS: Risks of extremely, very, and moderately preterm birth increased with BMI, and the highest risk was observed for obese women with extremely preterm birth (OR 3.43, 95% CI 1.07-10.97). Maternal obesity was significantly associated with spontaneous preterm labor (OR 1.98; 95% CI 1.13-3.47), premature rupture of the membranes (OR 2.04; 95% CI 1.08-3.86) and medically indicated preterm birth (OR 2.05; 95% CI 1.25-3.37). GDM and PE mediated 13.41 and 36.66% of the effect of obesity on preterm birth, respectively. GDM mediated 32.80% of the effect of obesity on spontaneous preterm labor and PE mediated 64.31% of the effect of obesity on medically indicated preterm birth. CONCLUSIONS: Maternal prepregnancy obesity was associated with all phenotypes of preterm birth, and the highest risks were extremely preterm birth and medically indicated preterm birth. GDM and PE partially mediated the association between obesity and preterm birth.

Association between the cut-off value of the first trimester fasting plasma glucose level and gestational diabetes mellitus: a retrospective study from southern China.

PURPOSE: Our previous studies have suggested that the first trimester fasting plasma glucose (FPG) level is associated with gestational diabetes mellitus (GDM) and is a predictor of GDM. The aim of the present study was to provide valuable insights into the accuracy of the first trimester FPG level in the screening and diagnosis of GDM in southern China. METHODS: This retrospective study included pregnant women who had their first trimester FPG level recorded at 9-13+6 weeks and underwent screening for GDM using the 2-h 75 g oral glucose tolerance test (OGTT) between the 24th and 28th gestational weeks. Differences between the GDM and non-GDM groups were assessed by Student's t test and the chi-squared test according to the nature of the variables. A restricted cubic spine was used to explore the relationship between the first trimester FPG level and the odds ratio (OR) of GDM in pregnant women. Cut-off values of first trimester FPG were determined using receiver operating characteristic (ROC) curves and the area under the curve (AUC), and 95% confidence intervals (CIs), the positive predictive value (PPV) and the negative predictive value (NPV) were calculated. RESULTS: The medical records of 28,030 pregnant women were analysed, and 4,669 (16.66%) of them were diagnosed with GDM. The average first trimester FPG level was 4.62 ± 0.37 mmol/L. The OR of GDM increased with increasing first trimester FPG levels and with a value of first trimester FPG of approximately 4.6 mmol/L, which was equal to 1 (Chi-Square = 665.79, P < 0.001), and then started to increase rapidly afterwards. The ROC curve for fasting plasma glucose in the first trimester (4.735 mmol/L) for predicting gestational diabetes mellitus in pregnant women was 0.608 (95% CI: 0.598-0.617), with a sensitivity of 0.490 and a specificity of 0.676. CONCLUSION: Based on the research, we recommend that all pregnant women undergo FPG testing in the first trimester, particularly at the first antenatal visit. Furthermore, we suggest that the risks of GDM should be given increased attention and management as soon as the first trimester FPG value is more than 4.7 mmol/L. First trimester FPG levels should be considered a screening marker when diagnosing GDM in pregnant women but this needs to be confirmed by more prospective studies. These factors may have a significant impact on the clinical treatment of pregnant women.

CMKLR1 deficiency attenuates androgen-induced lipid accumulation in mice.

Excess androgen-induced obesity has become a public health problem, and its prevalence has increased substantially in recent years. Chemokine-like receptor 1 (CMKLR1), a receptor of chemerin secreted by adipose tissue, is linked to adipocyte differentiation, adipose tissue development, and obesity. However, the effect of CMKLR1 signaling on androgen-mediated adiposity in vivo remains unclear. Using CMKLR1-knockout mice, we constructed an androgen-excess female mouse model through 5α-dihydrotestosterone (DHT) treatment and an androgen-deficient male mouse model by orchidectomy (ORX). For mechanism investigation, we used 2-(α-Naphthoyl) ethyltrimethylammonium iodide (α-NETA), an antagonist of CMKLR1, to suppress CMKLR1 in vivo and wortmannin, a PI3K signaling antagonist, to treat brown adipose tissue (BAT) explant cultures in vitro. Furthermore, we used histological examination and quantitative PCR, as well as Western blot analysis, glucose tolerance tests, and biochemical analysis of serum, to describe the phenotypes and the changes in gene expression. We demonstrated that excess androgen in the female mice resulted in larger cells in the white adipose tissue (WAT) and the BAT, whereas androgen deprivation in the male mice induced a reduction in cell size. Both of these adipocyte size effects could be attenuated in the CMKLR1-knockout mice. CMKLR1 deficiency influenced the effect of androgen treatment on adipose tissue by regulating the mRNA expression of the androgen receptor (AR) and adipocyte markers (such as Fabp4 and Cidea). Moreover, suppression of CMKLR1 by α-NETA could also reduce the extent of the adipocyte cell enlargement caused by DHT. Furthermore, we found that DHT could reduce the levels of phosphorylated ERK (pERK) in the BAT, while CMKLR1 inactivation inhibited this effect, which had been induced by DHT, through the PI3K signaling pathway. These findings reveal an antiobesity role of CMKLR1 deficiency in regulating lipid accumulation, highlighting the scientific importance for the further development of small-molecule CMKLR1 antagonists as fundamental research tools and/or as potential drugs for use in the treatment of adiposity.

Role of G protein-coupled receptor 1 in choriocarcinoma progression.

Choriocarcinoma is characterized by malignant proliferation and transformation of trophoblasts and is currently treated with systemic chemotherapeutic agents. The lack of specific targets for chemotherapeutic agents results in indiscriminate drug distribution. In our study, we aimed to delineate the mechanism by which G protein-coupled receptor 1 (GPR1) regulates the development of choriocarcinoma and thus investigated GPR1 as a prospective chemotherapeutic target. In this study, GPR1 expression levels were examined in several trophoblast cell lines. We found significantly higher GPR1 expression in choriocarcinoma cells (JEG3 and BeWo) than in normal trophoblast cells (HTR-8/SVneo). Additionally, we studied the role of GPR1 in choriocarcinoma in vitro and in vivo. GPR1 knockdown suppressed proliferation, invasion, and Akt and ERK phosphorylation in vitro and slowed tumor growth in vivo. Interestingly, GPR1 overexpression promoted increased proliferation, invasion, and Akt and ERK phosphorylation in vitro. Furthermore, we identified a specific GPR1-binding seven-amino acid peptide, LRH7-G3, that might also suppress choriocarcinoma in vitro and in vivo through phage display. Our study is the first to report that GPR1 may play a role in regulating choriocarcinoma progression through the Akt and ERK pathways. GPR1 could be a promising potential pharmaceutical target for choriocarcinoma.

Impact of GPR1 signaling on maternal high-fat feeding and placenta metabolism in mice.

Chemerin and G protein-coupled receptor 1 (GPR1) are increased in serum and placenta in mice during pregnancy. Interestingly, we observed increased serum chemerin levels and decreased GPR1 expression in placenta of high-fat-diet-fed mice compared with chow-fed mice at gestational day 18. GPR1 protein and gene levels were significantly decreased in gestational diabetes mellitus (GDM) patient placentas. Therefore, we hypothesized that chemerin/GPR1 signaling might participate in the pathogenic mechanism of GDM. We investigated the role of GPR1 in carbohydrate homeostasis during pregnancy using pregnant mice transfected with small interfering RNA for GPR1 or a negative control. GPR1 knockdown exacerbated glucose intolerance, disrupted lipid metabolism, and decreased ß-cell proliferation and insulin levels. Glucose transport protein-3 and fatty acid binding protein-4 were downregulated with reducing GPR1 in vivo and in vitro via phosphorylated AKT pathway. Taken together, our findings first demonstrate the expression of GPR1, the characterization of its direct biological effects in humans and mice, as well as the molecular mechanism that indicates the role of GPR1 signaling in maternal metabolism during pregnancy, suggesting a novel feedback mechanism to regulate glucose balance during pregnancy, and GPR1 could be a potential target for the detection and therapy of GDM.

Global mRNA and Long Non-Coding RNA Expression in the Placenta and White Adipose Tissue of Mice Fed a High-Fat Diet During Pregnancy.

BACKGROUND/AIMS: Gestational diabetes mellitus (GDM) is a common complication of pregnancy, but the mechanisms underlying the disorders remain unclear. The study aimed to identify mRNA and long non-coding RNA (lncRNA) profiles in placenta and gonadal fat of pregnant mice fed a high-fat diet and to investigate the transcripts and pathways involved in the development of gestational diabetes mellitus. METHODS: Deep and broad transcriptome profiling was performed to assess the expression of mRNAs and lncRNAs in placenta and gonadal fat from 3 mice fed an HFD and chow during pregnancy. Then, differentially expressed mRNAs and lncRNAs were validated by quantitative real-time PCR. The function of the differentially expressed mRNAs was determined by pathway and Gene Ontology (GO) analyses, and the physical or functional relationships between the lncRNAs and the corresponding mRNAs were determined. RESULTS: Our study revealed that 82 mRNAs and 52 lncRNAs were differentially expressed in the placenta of mice fed an HFD during pregnancy, and 202 mRNAs and 120 lncRNAs were differentially expressed in gonadal fat. GO and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed differentially expressed mRNAs of placenta were closely related to extracellular matrix interactions, digestion, adhesion, and metabolism, whereas the differentially expressed mRNAs in adipose tissue were related to metabolic and insulin signalling pathways. The gene network demonstrated that Actg2, Cnfn, Muc16, Serpina3k, NONMMUT068202, and NONMMUT068203, were the core of the network in placental tissue, and the genes Tkt, Acss2, and Elovl6 served as the core of the network in gonadal fat tissue. CONCLUSION: These newly identified key genes and pathways in mice might provide valuable information regarding the pathogenesis of GDM and might be used to improve early diagnosis, prevention, drug design, and clinical treatment.

Excessive gestational weight gain in accordance with the IOM criteria and the risk of hypertensive disorders of pregnancy: a meta-analysis.

BACKGROUND: Excessive gestational weight gain (GWG) is a potential risk factor for hypertensive disorders of pregnancy (HDP). METHODS: We systematically reviewed three electronic databases for relevant articles published in English: PubMed, EMBASE and Web of Science. The Newcastle-Ottawa Scale was used to assess study quality. Random-effects meta-analyses were performed to supply a pooled estimation of the OR comparing the risk of HDP among healthy pregnant women with and without excessive GWG. RESULTS: The pooled estimation for the association between excessive GWG and the risk of HDPs yielded an odds ratio (OR) of 1.79 (95% CI: 1.61-1.99). A subgroup analysis showed that women who had excessive GWG were more likely to have an HDP (OR 1.82; 95% CI 1.53-2.17), preeclampsia (OR 1.92; 95% CI 1.36-2.72), or gestational hypertension (OR 1.67; 95% CI 1.43-1.95). The pooled estimation for the association between excessive GWG and the risk of HDPs among pregestational normal weight women yielded an OR of 1.57 (95% CI 1.26-1.96). A subgroup analysis showed that women who had excessive GWG were more likely to have HDP (OR 1.45; 95% CI 1.09-1.92) or gestational hypertension (OR 1.51; 95% CI 1.22-1.86). The summary ORs of pre-gestational underweight women and pre-gestational overweight and obese women were 2.17 (95% CI 1.56-3.02) and 1.32 (95% CI 1.08-1.63), respectively. CONCLUSIONS: The findings of this study suggest that excessive GWG in accordance with the IOM recommendations influences the rate of HDP.

Clustering of metabolic risk factors and adverse pregnancy outcomes: a prospective cohort study.

BACKGROUND: The relative contributions of a cluster of metabolic risk factors to pregnancy complications are not fully understood. We investigated the correlation between clustering of metabolic risk factors and adverse pregnancy outcomes. METHODS: This prospective cohort study was performed on pregnant women who sought health care during their whole gestation in a women's and children's hospital. The pregnancy outcomes were also followed. Pre-pregnancy overweight/obesity, as well as pregnancy high triglycerides, low high-density lipoprotein-cholesterol, hyperglycemia and raised blood pressure were defined as metabolic risk factors. Adverse pregnancy outcomes included preterm delivery, small/large for gestational age, preeclampsia, gestational diabetes mellitus, neonatal asphyxia and foetal demise. Stratified analyses were conducted on a total of 5535 women according to classification in each metabolic risk factor. The adjusted odds ratio (OR) for adverse pregnancy outcomes according to the number of clustering metabolic factors was calculated using the logistic regression analysis. RESULTS: The number of metabolic risk factors and adverse pregnancy outcomes were positively correlated (Ptrend < 0.001). Compared with women without a metabolic risk factor, women with one metabolic risk factor had a risk (OR = 1.67 95%CI 1.42-1.96) of adverse pregnancy outcomes. Women with a cluster of two metabolic risk factors tended to develop more adverse pregnancy outcomes (OR = 3.32 95% CI 2.69-4.10), and the risk was much higher in women with a cluster of three or more metabolic risk factors (OR = 10.40 95%CI 7.37-14.69). CONCLUSIONS: Pregnant women with a cluster of metabolic risk factors are more likely to have adverse pregnancy outcomes. Copyright © 2016 John Wiley & Sons, Ltd.

Impact of preeclampsia on megakaryocytopoesis and platelet homeostasis of preterm infants.

UNLABELLED: The aim of this article is to investigate the megakaryopoyesis and thrombopoiesis in preterm infants born to mothers with preeclampsia and the potential effects mediated by soluble fms-like tyrosine kinase 1 (sFlt1) and thrombopoietin (TPO). A perspective case-control study was performed on 26 cord blood of preterm newborns born to mothers with preeclampsia (PE group) and 26 of preterms born to mothers without preeclampsia (control group). Circulating megakaryocyte count and megakaryocyte colony forming units (CFU-MK) were quantified by whole blood infiltration method and plasma clot culture system, respectively. Platelet activation markers, CD62P and CD63, were estimated by flow cytometry. Immunosorbent assays (ELISA) were employed to estimate plasma levels of sFlt1 and TPO of the two groups. When compared to the controls, infants born to mothers with PE had significantly lower peripheral platelet count (PE vs. CONTROLS: 157.9 [44.6] vs. 239.6 [57.5] × 10(9)/l, p < 0.001), circulating MK count (5.8 [1.0] vs. 7.7 [0.9]/ml, p < 0.001) and CFU-MK (14.1 [2.1] vs. 20.1 [2.8]/1 × 10(5) cell, p < 0.001); greater expressions of CD62P (15.5 [2.3] vs. 11.4 [1.9]% platelets, p < 0.001) and CD63 (12.3 [2.4] vs. 9.0 [1.6]% platelets, p < 0.001); increased plasma Flt level (130.1 [8.0] vs. 97.7 [8.7] pg/ml, p < 0.001) and TPO level (129.5 [17.8] vs. 98.9 [11.8] pg/ml, p < 0.001). In PE group, sFlt instead of TPO showed a significantly negative relationship with platelet counts, CFU-MK and circulating MK count, a positive relationship with CD62P, CD63 expressions. In control group, both sFlt and TPO did not show any relationship with these parameters. sFlt played important role in megakaryocytopoesis and platelet homeostasis in preterm infants born to mothers with PE. Its mechanism maybe the effect of impaired megakaryocyte formation and increased platelet activation.

Lipoprotein-associated phospholipase A2 is associated with postpartum hypertension in women with history of preeclampsia.

Both hypertension and preeclampsia (PE) are considered as inflammatory diseases. Lipoprotein-associated phospholipase A2 (Lp-PLA2) is an inflammatory marker associated with lipid metabolism. We aimed to study the correlation and predictive value of Lp-PLA2 in postpartum hypertension after PE. A group of 160 PE patients (PE group) and a separate group of 160 normal pregnant women (control group) were recruited from January 2010 to October 2011. The average age in the PE group was 28.4 ± 4.5 years and the average gestational age was 34.7 ± 1.1 weeks. The average age in the control group was 27.8 ± 4.5 years and the average gestational age was 35.5 ± 1.2 weeks. General information (including age, gestational age, parity, history of metabolic disease, family history of high blood pressure, height, body weight before childbirth, and blood pressure) and blood samples were collected for measuring Lp-PLA2 and lipid parameters. From February to April in 2013, 153 cases in the PE group and 132 in the control group were re-called. We assessed their postpartum health, pregnancy, height, weight, and blood pressure. Serum mass of Lp-PLA2 in the PE group (210.67 ± 17.98 ng/mL) was significantly higher compared with that in the control group (174.72 ± 30.26 ng/mL) (P < 0.01). The pro-gestation BMI, systolic blood pressure (SBP), diastolic blood pressure, total cholesterol, triglyceride, and low-density lipoprotein-cholesterol (LDL-C) were also significantly higher. Correlation analysis showed that the level of Lp-PLA2 and SBP (r = 0.31), LDL-C (r = 0.37) were positively correlated. The incidence of postpartum hypertension in the PE group was higher than that in the normal control group. Logistic regression analysis showed that prenatal Lp-PLA2 mass was an independent risk factor for PE postpartum hypertension (OR 1.134,95 % CI 1.086-1.185). ROC curve analysis showed that the sensitivity of predicting postpartum hypertension was 73.2% and the specific degree was 86.6%, with Lp-PLA2 level of 217.75 ng/mL for boundary value. The onset of postpartum hypertension in PE patients may contribute to vascular inflammation, which is associated with antepartum lipid metabolism.

Lipoprotein-associated phospholipase A2 and AGEs are associated with cardiovascular risk factors in women with history of gestational diabetes mellitus.

OBJECTIVE: Although most women with gestational diabetes mellitus (GDM) return to normal glucose tolerance after delivery, they have increased risk of cardiometabolic diseases. This study aimed to evaluate the relationships between plasma levels of Lp-pla2 and AGEs and cardiometabolic risk factors in women with GDM. METHODS: 190 women with GDM (cases) and 80 healthy women (controls) were enrolled. Demographic and clinical data were collected and analyzed about 2 years after the delivery. RESULTS: Of the 190 cases, 19 (10%), 38 (20%) and 10 (5%) had type 2 diabetes mellitus, metabolic syndrome and hypertension after delivery, respectively. There were significant differences in variables between cases and controls: Lp-pla2 (pg/mL) 1991.5 ± 905.3 versus 1527.0 ± 799.8; AGEs (ng/mL) 403.0 ± 208.6 versus 321.8 ± 150.3. The plasma Lp-pla2 and AGEs levels were positively correlated with metabolic indexes in women with previous GDM. CONCLUSION: Women with GDM have increased risk of cardiometabolic disease. AGEs and Lp-pla2 could be utilized as novel biomarkers to identify at an early stage of women with increased risk of metabolic and cardiovascular disease.

[Incident and related risk factors of hypertension in women with a history of preeclampsia].

OBJECTIVE: To investigate the prevalence of hypertension in women with a history of preeclampsia (PE) and to estimate related risk factors. METHODS: In this prospective case-control study, we collected clinical data from 809 women with a history of PE and 3 421 women with normal pregnancy from January 2008 to June 2012. Between November 2012 and April 2013, 651 women in PE group and 2 684 women with normal pregnancy group were recruited at the same time for collecting postpartum data including blood pressure, blood glucose and blood lipid. Binary logistic regression analysis was applied to analyze the relative factors of postpartum blood pressure. RESULTS: Prevalence of hypertension in PE group was higher than those with normal pregnancy (17.2% (112/651) vs. 1.1% (30/2 684), P < 0.01). Prevalence of hypertension in severe PE and mild PE patients was similar (20.1% (58/289) vs. 15.2% (55/362), P = 0.103). Binary logistic regression analysis indicated that progestational body mass index (OR = 1.379, 95% CI: 1.257-1.510, P < 0.05) , antepartum systolic blood pressure (OR = 1.025, 95%CI:1.012-1.040, P < 0.05) , antepartum triglyceride (OR = 1.002, 95% CI: 1.002-1.410, P < 0.05) , antepartum fasting blood glucose (OR = 1.733, 95% CI: 1.047-2.870, P < 0.05) , postpartum body mass index (OR = 1.279, 95% CI: 1.199-1.363, P < 0.05), postpartum fasting insulin (OR = 1.107, 95% CI: 1.055-1.162, P < 0.05) , systolic blood pressure difference between antepartum and postpartum (OR = 1.024, 95% CI :1.011-1.037, P < 0.05) , difference on triglyceride value between antepartum and postpartum (OR = 1.26, 95% CI: 1.069-1.486, P < 0.01), difference value of HOMA-IR between antepartum and postpartum (OR = 2.448, 95% CI: 1.330-4.500, P < 0.01) and difference value of high density lipoprotein cholesterol between antepartum and postpartum (OR = 1.699, 95% CI: 1.277-2.260, P < 0.05) were associated with hypertension after pregnancy. CONCLUSIONS: Women with history of PE are associated with higher risk of postpartum hypertension. Increased blood pressure, abnormal glucose and lipid metabolism during pregnancy are major risk factors for postpartum hypertension.

Characterizing immune variation and diagnostic indicators of preeclampsia by single-cell RNA sequencing and machine learning.

Preeclampsia is a multifactorial and heterogeneous complication of pregnancy. Here, we utilize single-cell RNA sequencing to dissect the involvement of circulating immune cells in preeclampsia. Our findings reveal downregulation of immune response in lymphocyte subsets in preeclampsia, such as reduction in natural killer cells and cytotoxic genes expression, and expansion of regulatory T cells. But the activation of naïve T cell and monocyte subsets, as well as increased MHC-II-mediated pathway in antigen-presenting cells were still observed in preeclampsia. Notably, we identified key monocyte subsets in preeclampsia, with significantly increased expression of angiogenesis pathways and pro-inflammatory S100 family genes in VCAN+ monocytes and IFN+ non-classical monocytes. Furthermore, four cell-type-specific machine-learning models have been developed to identify potential diagnostic indicators of preeclampsia. Collectively, our study demonstrates transcriptomic alternations of circulating immune cells and identifies immune components that could be involved in pathophysiology of preeclampsia.

Simultaneous removal of nitrogen and phosphorus from wastewater by means of FeS-based autotrophic denitrification.

The efficacy of iron(II) sulfide (FeS)-based autotrophic denitrification in simultaneous nitrogen and phosphorus removal from wastewater was studied with batch experiments. It was efficient at a wide pH range of 5-9, and temperature range of 10-40 °C. The concentrations of NH4⁺-N, Mg²âº and HCO3⁻ in the wastewater should be kept over 7.8, 0.24 and 30 mg L⁻¹ for efficient nitrate (NO3⁻-N) reduction, respectively. The NO3⁻-N removal rate increased from 0 to 82 mg L⁻¹ d⁻¹ and then leveled off when the NO3⁻-N concentration increased from 0 to 415 mg L⁻¹ and then to 700 mg L⁻¹, respectively. The NO3⁻-N removal rate quickly increased, leveled off, and then sharply decreased when the PO4³â»-P concentration increased from 0 to 0.1 mg L⁻¹, then to 114.0 mg L⁻¹, and further to 683.8 mg L⁻¹, respectively. The PO4³â»-P removal was over 98% when the PO4³â»-P concentration ranged 0-683.3 mg L⁻¹. During treatment of the secondary effluent of a local municipal wastewater treatment plant containing NO3⁻-N of 14.9 mg L⁻¹ and total phosphorus (TP) of 3.9 mg L⁻¹, NO3⁻-N was reduced to 1.1 mg L⁻¹ and TP was completely removed.