RNA-binding protein SORBS2 increases human trophoblast cell migration via stabilizing HK2 mRNA in preeclampsia.

In brief: SORBS2, an RNA-binding protein, is identified as a regulator of aerobic glycolysis, which is essential for trophoblast migration and placental development. Reduced SORBS2 expression in preeclampsia may impair trophoblast migration by affecting mRNA stability and glycolysis, suggesting its role in the disease's pathogenesis. Abstract: Insufficient trophoblast migration and impaired uterine spiral artery remodeling are implicated in the pathogenesis of preeclampsia, contributing to inadequate placentation. However, the molecular mechanism underlying this process remains unclear. Aerobic glycolysis, which produces substantial lactate, is crucial for establishing a favorable microenvironment for early uterine preparation and supporting embryo implantation and trophoblast migration. In the present study, we have demonstrated that SORBS2, an RNA-binding protein, regulated aerobic glycolysis and significantly improved trophoblast migration in vitro. Our results showed that SORBS2 expression was significantly reduced in human PE placentas and trophoblasts during hypoxia. Overexpression of SORBS2 enhanced cell proliferation and migration, whereas knockdown of SORBS2 decreased these functions in HTR-8/SVneo cells. Mechanistic studies have demonstrated that SORBS2 directly interacts with the 3' untranslated regions (UTRs) of key glycolysis-related genes, specifically HK2. This interaction results in enhanced stability of HK2 and activation of glycolysis. Moreover, silencing HK2 abrogated the enhancement of proliferation and migration of HTR-8/SVneo cells induced by SORBS2. In conclusion, our findings suggest that the downregulation of SORBS2 may contribute to the pathogenesis of preeclampsia by regulating mRNA stability and inhibiting trophoblast migration during placentation.

Downregulation of miR-126-3p expression contributes to increased inflammatory response in placental trophoblasts in preeclampsia.

MiR-126-3p is a prototype of an endothelial miRNA and has protective effects on endothelial cells. However, little is known about the effects of miR-126-3p on placental trophoblasts. In the present study, we tested the hypothesis that aberrant miR-126-3p expression is present in preeclamptic placenta which contributes to increased inflammatory response in trophoblasts. Placentas were obtained immediately after delivery from normotensive and preeclamptic pregnancies. Villous tissue was either fixed with formalin or used for trophoblast isolation. Trophoblast miR-126-3p expression was assessed by in situ hybridization of formalin-fixed tissue sections and by RT-PCR in cultured syncytiotrophoblasts. Culture medium was collected for measurement of IL-6, TNFα, and 8-Isoprostane production by ELISA and total cellular protein was collected for evaluation of HIF1α expression by Western blot. Effects of overexpression of miR-126-3p in trophoblasts on cytokine production were tested by transfection of pre-mir-126, a precursor of miR-126, into primary isolated trophoblasts. We found that downregulation of miR-126-3p expression was associated with increased IL-6 and TNFα production in trophoblasts from preeclamptic placentas vs. normal placentas. Moreover, transient overexpression of miR-126-3p significantly reduced IL-6 and TNFα production in trophoblasts from both normal and preeclamptic placentas. We further found that increase in miR-126-3p expression not only suppressed hypoxia-induced increases in IL-6 and TNFα production, but also attenuated hypoxia-induced increases in HIF1α expression and 8-Isoprostane production in trophoblasts cultured under hypoxic condition. These results provide plausible evidence that downregulation of miR-126-3p expression reduces anti-inflammatory and anti-oxidative stress activities in placental trophoblasts in preeclampsia.

Upregulation of METTL3 expression and m6A RNA methylation in placental trophoblasts in preeclampsia.

INTRODUCTION: N6-methyladenosine (m6A) has been recognized as one of the most abundant and functionally relevant modifications of RNAs and plays critical roles in biological and pathological processes. Placental trophoblast dysfunction significantly contributes to the pathogenesis of preeclampsia. The present study aimed to determine if altered m6A expression occurs in placental trophoblasts in preeclampsia. Expression of m6A methyltransferase (methyltransferase like 3 (METTL3)), m6A demethylases (fat mass and obesity-associated protein (FTO) and AlkB homolog 5 (ALKBH5)), and m6A reader protein, heterogeneous nuclear ribonucleoprotein C1/C2 (hnRNPC1/C2), were also examined. METHODS: A total of 43 placentas (20 normal term, 5 normotensive preterm, and 18 preeclamptic) were used in the study. Expression of m6A, METTL3, FTO, ALKBH5, and hnRNPC1/C2 were examined by immunostaining in villous tissue sections and/or by Western blot of total cellular protein in trophoblasts isolated from normotensive and preeclamptic placentas. Total RNA extracted from trophoblasts was used to measure m6A RNA methylation. Effects of METTL3 on m6A RNA methylation and hnRNPC1/C2 expression were assessed by transfection of METTL3 siRNA in trophoblasts from preeclamptic placentas. RESULTS: Expression of m6A and m6A RNA methylation were significantly increased in trophoblasts from preeclamptic vs. normotensive placentas, p < 0.05. Expression of METTL3 and hnRNPC1/C2, but not FTO and ALKBH5, was significantly upregulated in trophoblasts from preeclamptic vs. normotensive placentas, p < 0.01. Transfection of METTL3 siRNA significantly reduced the level of m6A RNA methylation and hnRNPC1/C2 expression in trophoblasts from preeclamptic placentas, p < 0.05. CONCLUSION: The finding of increased METTL3 expression and m6A RNA methylation associated with increased hnRNPC1/C2 expression provides a new posttranscriptional mechanism that aberrant m6A modification may contribute to trophoblast dysfunction in preeclampsia.

Upregulation of histone H3K9 methylation in fetal endothelial cells from preeclamptic pregnancies.

Adverse intrauterine environment has been considered a predisposing factor for fetal programming in preeclampsia. Using human umbilical vein endothelial cells (HUVECs), we specifically explored if aberrant histone methylation occurs in fetal endothelial cells in preeclampsia. Strikingly, we found that increased di-, and tri-methylation of histone H3 lysine 9 (H3K9me2 and H3K9me3) expression were associated with upregulation of methyltransferase G9a and downregulation of endothelial nitric oxide synthase and CuZn-SOD expression in preeclamptic HUVECs. We further demonstrated that hypoxia-induced hypermethylation of H3K9 and reduced CuZn-SOD expression mimicked what were seen in preeclamptic HUVECs and inhibition of G9a could attenuate these hypoxia-induced adverse events. Our study was the first to identify hypermethylation status in fetal endothelial cells in preeclampsia, which provides plausible evidence that increased oxidative stress in the intrauterine environment is likely a mechanism to induce aberrant histone modification in fetal endothelial cells which may have a significant impact on fetal programming in preeclampsia.

Identification and characterization of methylation-mediated transcriptional dysregulation dictate methylation roles in preeclampsia.

BACKGROUND: Preeclampsia (PE) is a heterogeneous, hypertensive disorder of pregnancy, with no robust biomarkers or effective treatments. PE increases the risk of poor outcomes for both the mother and the baby. Methylation-mediated transcriptional dysregulation motifs (methTDMs) could contribute the PE development. However, precise functional roles of methTDMs in PE have not been globally described. METHODS: Here, we develop a comprehensive and computational pipeline to identify PE-specific methTDMs following TF, gene, methylation expression profile, and experimentally verified TF-gene interactions. RESULTS: The regulation patterns of methTDMs are multiple and complex in PE and contain relax inhibition, intensify inhibition, relax activation, intensify activation, reverse activation, and reverse inhibition. A core module is extracted from global methTDM network to further depict the mechanism of methTDMs in PE. The common and specific features of any two kinds of regulation pattern are also analyzed in PE. Some key methylation sites, TFs, and genes such as IL2RG are identified in PE. Functional analysis shows that methTDMs are associated with immune-, insulin-, and NK cell-related functions. Drug-related network identifies some key drug repurposing candidates such as NADH. CONCLUSION: Collectively, the study highlighted the effect of methylation on the transcription process in PE. MethTDMs could contribute to identify specific biomarkers and drug repurposing candidates for PE.

Circulating lncRNA BC030099 Increases in Preeclampsia Patients.

Long noncoding RNAs (lncRNAs) have increasingly been shown to be important biological regulators involved in numerous diseases. Further, increasing evidence demonstrates that circulating lncRNAs can be used as diagnostic biomarkers. Therefore, the purpose of this study was to evaluate the potential for circulating lncRNAs as novel biomarkers for the diagnosis of preeclampsia. In the present study, we measured the expression of five lncRNAs known to be relevant to the uterus in whole blood samples from 48 preeclampsia patients and 24 non-preeclampsia healthy subjects using qRT-PCR. We found that circulating levels of lncRNA BC030099 were significantly higher in patients with preeclampsia (1.232 ± 0.4870) than in non-preeclampsia healthy subjects (0.9928 ± 0.2008, p < 0.05). The area under the receiver operating characteristic (ROC) curve for lncRNA BC030099 was 0.713. Univariate and multivariate analyses identified lncRNA BC030099 as an independent predictor for preeclampsia. In brief, our results suggest that increased plasma levels of lncRNA BC030099 are associated with an increased risk of preeclampsia and may be considered a novel biomarker.

Aromatic alkyne insertion into six-membered cyclometalated pyridine complexes of iridium: different insertion modes and structurally novel products.

Reactions of 2-benzoylpyridine or 2-benzylpyridine with [Cp*MCl2]2 (M = Ir, Rh) have been carried out in the presence of NaOAc in refluxing methanol, which form the corresponding six-membered cyclometalated products (1-3) except for the reaction of 2-benzylpyridine with [Cp*RhCl2]2. Insertion reactions of two six-membered cyclometalated pyridine iridium complexes (1 and 2) with terminal or internal aromatic alkynes were studied. Terminal alkynes p-XC6H4C[triple bond, length as m-dash]CH (X = H, MeO, and F) with 1 give the corresponding five- and seven-membered doubly cycloiridated complexes 4a-c, internal alkynes p-XC6H4C[triple bond, length as m-dash]CC6H4X-p (X = H, MeO, and Br) form the similar five- and seven-membered doubly cycloiridated complexes (5a,b) and/or di-insertion products (6a,c), whereas the acyl alkyne PhC[triple bond, length as m-dash]CCOPh affords the novel spiro-metalated complex 7. For complex 2, internal alkynes p-XC6H4C[triple bond, length as m-dash]CC6H4X-p (X = H, MeO, and Br) form similar five- and seven-membered doubly cycloiridated complexes (8a-c). However, in the case of PhC[triple bond, length as m-dash]CCOPh, the reaction gives the novel four-membered cyclometalated complex 9. These results suggest that the products formed by alkyne insertion reactions of the six-membered cycloiridated pyridine complexes are very diverse. Plausible pathways for the formation of these novel insertion products were proposed. Molecular structures of seven cyclometalated complexes were determined by X-ray diffraction.

Synthesis, Structures, and Reactivity of Single and Double Cyclometalated Complexes Formed by Reactions of [Cp*MCl2]2 (M = Ir and Rh) with Dinaphthyl Phosphines.

Reactions of two dinaphthyl phosphines with [Cp*IrCl2]2 have been carried out. In the case of di(α-naphthyl)phenylphosphine (1a), a simple P-coordinated neutral adduct 2a is obtained. However, tert-butyldi(α-naphthyl)phenylphosphine (1b) is cyclometalated to form [Cp*IrCl(P^C)] (3b). Complexes 2a and 3a undergo further cyclometalation to give the corresponding double cyclometalated complexes [Cp*Ir(C^P^C)] (4a,b) upon heating. In the presence of sodium acetate, reactions of 1a,b with [Cp*IrCl2]2 directly afford the final double cyclometalated complexes (4a,b). In the absence of acetate, [Cp*RhCl2]2 shows no reaction with 1a,b, whereas with acetate the reactions form the corresponding single cyclometalated complexes [Cp*RhCl(P^C)] (5a,b), which react with t BuOK to form the corresponding rhodium hydride complexes (6a,b). Treatment of 4a with CuCl2 or I2 leads to opening of two Ir-C σ bonds to yield the corresponding P-coordinated iridium dihalide (7 or 8) by means of an intramolecular C-C coupling reaction. A new chiral phosphine (11) is formed by the ligand-exchange reaction of 8 with PMe3. Reactions of the single cycloiridated complex 3b with terminal aromatic alkynes result in the corresponding five- and six-membered doubly cycloiridated complex 12 and/or η2-alkene coordinated complexes 13-15; the latter discloses that the electronic effect of terminal alkynes affects the regioselectivity. While the single cyclorhodated complex 5b reacts with terminal aromatic alkynes to form the corresponding six-membered cyclometalated complexes 16a-c by vinylidene rearrangement/1,1-insertion. Plausible pathways for formation of insertion products 13-16 were proposed. Molecular structures of twelve new complexes were determined by X-ray diffraction.

Genome sequencing of strain Cellulosimicrobium sp. TH-20 with ginseng biotransformation ability.

Biotransformation for increasing the pharmaceutical effect of ginsenosides is getting more and more attractions. Strain Cellulosimicrobium sp. TH-20 isolated from ginseng soil samples was identified to produce enzymes contributing to its excellent biotransformation activity against ginsenosides, the main active components of ginseng. Based on phylogenetic tree and homology analysis, the strain can be designated as Cellulosimicrobium sp. Genome sequencing was performed using the Illumina Miseq to explore the functional genes involved in ginsenoside transformation. The draft genome of Cellulosimicrobium sp. TH-20 encoded 3450 open reading frames, 51 tRNA, and 9 rRNA. All ORFs were annotated using NCBI BLAST with non-redundant proteins, Gene Ontology, Cluster of Orthologous Gene, and Kyoto Encyclopedia of Genes and Genomes databases. A total of 11 genes were selected based on the functional annotation analysis. These genes are relevant to ginsenoside biotransformation, including 6 for beta-glucosidase, 1 for alpha-N-arabinofuranosidase, 1 for alpha-1,6-glucosidase, 1 for endo-1,4-beta-xylanase, 1 for alpha-L-arabinofuranosidase, and 1 for beta-galactosidase. These glycosidases were predicted to catalyze the hydrolysis of sugar moieties attached to the aglycon of ginsenosides and led to the transformation of PPD-type and PPT-type ginsenosides.

Enrichment Analysis Identifies Functional MicroRNA-Disease Associations in Humans.

Substantial evidence has shown that microRNAs (miRNAs) may be causally linked to the occurrence and progression of human diseases. Herein, we conducted an enrichment analysis to identify potential functional miRNA-disease associations (MDAs) in humans by integrating currently known biological data: miRNA-target interactions (MTIs), protein-protein interactions, and gene-disease associations. Two contributing factors to functional miRNA-disease associations were quantitatively considered: the direct effects of miRNA that target disease-related genes, and indirect effects triggered by protein-protein interactions. Ninety-nine miRNAs were scanned for possible functional association with 2223 MeSH-defined human diseases. Each miRNA was experimentally validated to target ≥ 10 mRNA genes. Putative MDAs were identified when at least one MTI was confidently validated for a disease. Overall, 19648 putative MDAs were found, of which 10.0% was experimentally validated. Further results suggest that filtering for miRNAs that target a greater number of disease-related genes (n ≥ 8) can significantly enrich for true MDAs from the set of putative associations (enrichment rate = 60.7%, adjusted hypergeometric p = 2.41×10-91). Considering the indirect effects of miRNAs further elevated the enrichment rate to 72.6%. By using this method, a novel MDA between miR-24 and ovarian cancer was found. Compared with scramble miRNA overexpression of miR-24 was validated to remarkably induce ovarian cancer cells apoptosis. Our study provides novel insight into factors contributing to functional MDAs by integrating large quantities of previously generated biological data, and establishes a feasible method to identify plausible associations with high confidence.

Analysis on the outcomes of hepatitis B virus perinatal vertical transmission: nested case-control study.

OBJECTIVE: Hepatitis B virus (HBV) infection is a public health problem worldwide, with vertical transmission as the leading transmission route. Therefore, it is very important to explore the risk factors associating with HBV perinatal transmission, providing valuable information for preventive and curative means for HBV perinatal infections. In this study, we systematically investigated the impact of adverse outcomes of HBV maternal infection on fetal intrauterine infection. PATIENTS AND METHODS: This is a nested case-control study that included 42 hepatitis B surface antigen (HBsAg)-positive pregnant women. Gestational weeks, delivery modes, alanine aminotransferase levels, platelet counts, liver ultrasonography results as well as adverse pregnancy outcomes for the pregnant mothers and Apgar scores at both 1 and 5 min, birth weight, and height for the newborn infants were recorded. Vein blood from pregnant mothers and cord blood immediately after delivery were collected for the detection of HBsAg, antibodies to hepatitis B surface antigen, hepatitis B e antigen (HBeAg), antibody to hepatitis B e antigen, hepatitis B core antigen, and HBV DNA. Placental tissues were collected for detection of HBV DNA. RESULTS: Positive HBeAg as well as HBV DNA in the mother's serum were correlated closely with HBV intrauterine infection. Mother's age, delivery mode, alanine aminotransferase, blood platelet count, clinical HBV infection features, premature labor, gestational diabetes mellitus, pre-eclampsia, fetal growth retardation, fetal distress, Apgar scores of the infant as well as the HBV infection status of the placenta all failed to show a statistically significant correlation with intrauterine infection. CONCLUSION: High level of HBV in maternal blood was one of the risk factors accounting for intrauterine infection.

MicroRNA-650 expression in glioma is associated with prognosis of patients.

MicroRNAs are known as non-coding RNAs that regulate the expression of target mRNA. Accumulating evidence has indicated that microRNA expression in human malignancies can be utilized as a prognostic marker for patients. However, the prognostic value of miR-650 in human glioma has not been investigated yet. In the present investigation, we have recruited 168 cases glioma specimens and 21 normal control brain specimens. Quantitative real-time PCR was carried out to investigate the expression of miR-650. Kaplan-Meier analysis and Cox's proportional hazards model was used to evaluate the association of miR-650 with prognosis of glioma patients. Results showed that miR-650 expression was increased in glioma compared with normal control specimens (P < 0.001). It was also found that miR-650 expression was related to World Health Organization grade and Karnofsky performance score (KPS) for high expression was more frequently detected in glioma of high grade or low KPS score (P < 0.001). The prognosis of glioma with high miR-650 expression was significantly worse compared with that of glioma with low miR-650 expression. These results proved that miR-650 expression was a significant prognostic indicator in glioma, which may suggest new management of human glioma.

Polysaccharides from Bupleurum chinense impact the recruitment and migration of neutrophils by blocking fMLP chemoattractant receptor-mediated functions.

Although neutrophils play important roles in host defense, sometimes they contribute to inflammation-related tissue injuries. Thus, restriction of their recruitment to the inflammatory sites is a promising strategy in the amelioration of inflammatory disease. The previous studies reported the anti-inflammatory effects of Bupleurum chinense, but the active ingredients and possible mechanism are still unclear. Here, we isolated water-soluble polysaccharides (BCPs) from B. chinense, to evaluate its anti-inflammatory effects and possible mechanism. The present results showed that BCPs significantly impaired the in vivo neutrophil infiltration, as well as the migration capacity of dHL-60 cells in vitro. In addition, BCPs inhibits chemoattractant fMLP-induced activation and clustering of ß2 integrin. BCPs impacted fMLP-induced actin polymerization and the activation of cytoskeleton regulatory molecules, Vav1 and Rac1. Together, BCPs significantly impacted recruitment and migration of neutrophils by blocking chemoattractant receptor-mediated functions, and it possesses a potential as novel anti-inflammatory drug.

Decreased expression of NDRG2 is related to poor overall survival in patients with glioma.

Members of the NDRG (N-Myc downstream-regulated) gene family have been shown to play a variety of roles in human malignancies. In the present study, we examined the expression of NDRG2 protein in glioma samples of WHO grades I-IV. We also investigated the association between NDRG2 expression and survival. Immunohistochemical analysis was used to measure NDRG2 protein expression in 316 specimens of human glioma and 41 normal control tissues. Survival analysis was performed using the Kaplan-Meier method and Cox's proportional hazards model. We found that NDRG2 expression was reduced in glioma relative to normal tissue, and that NDRG2 expression decreased with increasing glioma grade. Kaplan-Meier analysis showed that patients without NDRG2 expression had a lower survival rate than other patients. Multivariate analysis showed that NDRG2 expression was an independent prognostic factor for overall survival of patients with glioma. The present study provides the first evidence that NDRG2 expression is decreased in gliomas, indicating that NDRG2 may play an inhibitory role during the development of gliomas. NDRG2 expression may also be a significant and independent prognostic indicator for glioma.

Decreased expression of NDRG1 in glioma is related to tumor progression and survival of patients.

The aim of the study was to examine the expression of NDRG1 gene in glioma samples with different WHO grades and its association with survival. About 168 glioma specimens and 21 normal control tissues were collected. Immunochemistry assay, quantitative real-time PCR and Western blot analysis were carried out to investigate the expression of NDRG1 and Myc. Kaplan-Meier method and Cox's proportional hazards model were used in survival analysis. Immunohistochemistry showed that Ndrg1 expression was reduced in glioma. NDRG1 mRNA and protein levels were lower in glioma compared to control on real-time PCR and Western blot analysis (P < 0.001). Its expression levels increase from grade IV to grade I glioma on real-time PCR, immunohistochemistry analysis (P < 0.001) and Western blot. On the contrary, the expression of Myc by real-time PCR and Western blot showed the opposite trend of NDRG1. The survival rate of Ndrg1-negative patients was lower than that of Ndrg1-positive patients. We confirmed that the loss of NDRG1 expression was a significant and independent prognostic indicator in glioma by multivariate analysis. NDRG1 may play an inhibitory role during the development of glioma and may be a potential prognosis predictor of glioma.