Monitoring of hemostatic parameters for early prediction of first-trimester miscarriage.

BACKGROUND: Hypercoagulation starts as early as the first-trimester pregnancy and is a risk factor for thromboembolic events which are associated with miscarriage. Our study aimed to investigate coagulation, platelets, and fibrinolysis parameters alteration amongst trimester-specific normal pregnancy and first-trimester miscarriage patients. We also test the accuracy of haemostatic parameters determination for prediction of first-trimester miscarriage. METHODS: Retrospective investigation of 50 women whose most recent pregnancy had ended in the first trimester and 54 age-matched consecutive normal pregnancy between 2016 and 2019. Furthermore, 51 non-pregnant, age-matched women were included in parallel to healthy controls. Twelve screening tests for coagulation and platelet parameters were assessed. RESULTS: We found plasma levels of aPTT, FBG, and TT were significantly prolonged or decreased in miscarriage subjects than the corresponding first phase in normal pregnancies. PT, INR, aPTT, and d-dimer all shift back to normal in miscarriage patients compared with non-pregnant women. Shortened aPTT combined with TT and FBG can predicted the occurrence of first-trimester miscarriage with an AUC of 0.831. CONCLUSIONS: Routine assessment of aPTT combined with TT and FBG is a low-cost, widely available marker for prediction of first-trimester miscarriage.

A novel potentiometric immunoassay for carcinoma antigen 15-3 by coupling enzymatic biocatalytic precipitation with a nanogold labelling strategy.

Methods based on potentiometric measurement have been developed for immunoassays, but most exhibit low sensitivities and are unsuitable for early diagnosis of disease. Herein we design a new potentiometric immunosensing platform for the sensitive detection of carcinoma antigen 15-3 (CA 15-3) by coupling with enzymatic biocatalytic precipitation and a nanogold labeling technique. The sandwich-type immunoreaction is carried out on a monoclonal anti-CA 15-3 capture antibody-modified working electrode, using horseradish peroxidase (HRP) and polyclonal anti-CA 15-3 detection antibody-labeled gold nanoparticles. Upon the introduction of target CA 15-3, the carried HRP molecules with an immunocomplex catalyze the oxidation of 4-chloro-1-naphthol (4-CN) into the insoluble benzo-4-chlorohexadienone. The formed product coated on the surface of the modified electrode results in a change of the electrical potential. Under optimal conditions, the shift in the electrical potential relative to the background signal increases with the increasing target CA 15-3 concentration, and exhibits a good linear relationship within the dynamic range of 0.01-30 U mL-1 at a detection limit of 7.8 mU mL-1. Good precision and reproducibility, and high specificity were acquired for the analysis of 15 human serum specimens, giving well matched results with those obtained from a human CA 15-3 ELISA kit.

Sensitive electrochemical detection of microRNA-21 based on propylamine-functionalized mesoporous silica with glucometer readout.

A new homogeneous electrochemical sensing system was developed for sensitive detection of microRNA-21 (miRNA-21) based on target-induced glucose release from propylamine-functionalized mesoporous silica nanoparticle (MSN) with glucometer readout. Glucose molecules (as the signal tracers) were initially gated into the pores through the interaction of the negatively charged anchor DNA with the aminated MSN. Upon addition of target miRNA, the analyte competitively hybridized with anchor DNA to form the RNA-DNA duplex, thus resulting in detachment of anchor DNA from the MSN accompanying the pore opening. The loaded glucose molecules released out from the pores because of concentration gradients, which could be detected by using a portable personal glucometer (PGM). Experimental results indicated that the PGM signal increased with the increasing miRNA level, and exhibited a good linear dependence on the miRNA-21 concentration from 50 pM to 5.0 nM with a detection limit of 19 pM under optimum conditions. Additionally, multifunctional mesoporous silica nanoparticles also showed good stability and favorable selectivity, and satisfactory accuracy for the miRNA detection in cell lysates with quantitative real-time polymerase chain reaction (qRT-PCR). Such good analytical performance endows it as a promising scheme for the efficient and convenient detection of miRNA in clinical diagnosis and therapy. Graphical abstract An electrochemical sensing system is designed for detection of microRNA-21 based on target-induced glucose release from propylamine-functionalized mesoporous silica nanoparticle with glucometer readout.

Low-Density Lipoprotein Receptor Signaling Mediates the Triglyceride-Lowering Action of Akkermansia muciniphila in Genetic-Induced Hyperlipidemia.

OBJECTIVE: Akkermansia muciniphila (A muciniphila) is a mucin-degrading bacterium that resides in the mucus layer whose abundance inversely correlates with body weight and the development of diabetes mellitus in mice and humans. The objective of this study was to explore the regulatory effect of A muciniphila on host lipoprotein metabolism, insulin sensitivity, and hepatic metabolic inflammation. APPROACH AND RESULTS: By establishing a novel mouse model that colonized the A muciniphila in the gastrointestinal tract of the cAMP-responsive binding protein H (CREBH)-deficient mouse and in vivo chylomicron assay, we found that increased colonization of A muciniphila in the gastrointestinal tract of wild-type mice protected mice from an acute fat load-induced hyperlipidemia compared with vehicle-treated mice. A muciniphila administration also significantly ameliorated chronic hypertriglyceridemia, improved insulin sensitivity, and prevented overproduction of postprandial chylomicrons in CREBH-null mice. Mechanistic studies revealed that increased A muciniphila colonization induced expression of low-density lipoprotein receptors and apolipoprotein E in the hepatocytes of CREBH-null mice, which facilitated the uptake of intermediate-density lipoprotein via the mediation of apolipoprotein B100 and apolipoprotein E, leading to the increased clearance of triglyceride-rich lipoprotein remnants, chylomicron remnants, and intermediate-density lipoproteins, from the circulation. Treatment with A muciniphila further improved hepatic endoplasmic reticulum stress and metabolic inflammation in CREBH-null mice. CONCLUSIONS: Increased colonization of the disease-protective gut bacteria A muciniphila protected the host from acute and chronic hyperlipidemia by enhancing the low-density lipoprotein receptor expression and alleviating hepatic endoplasmic reticulum stress and the inflammatory response in CREBH-null mice.

Hepatic mitochondrial and ER stress induced by defective PPARα signaling in the pathogenesis of hepatic steatosis.

Emerging evidence demonstrates a close interplay between disturbances in mitochondrial function and ER homeostasis in the development of the metabolic syndrome. The present investigation sought to advance our understanding of the communication between mitochondrial dysfunction and ER stress in the onset of hepatic steatosis in male rodents with defective peroxisome proliferator-activated receptor-α (PPARα) signaling. Genetic depletion of PPARα or perturbation of PPARα signaling by high-fructose diet compromised the functional activity of metabolic enzymes involved in mitochondrial fatty acid ß-oxidation and induced hepatic mitochondrial stress in rats and mice. Inhibition of PPARα activity further enhanced the expression of apolipoprotein B (apoB) mRNA and protein, which was associated with reduced mRNA expression of the sarco/endoplasmic reticulum calcium ATPase (SERCA), the induction of hepatic ER stress, and hepatic steatosis. Restoration of PPARα activity recovered the metabolic function of the mitochondria and ER, alleviated systemic hypertriglyceridemia, and improved hepatic steatosis. These findings unveil novel roles for PPARα in mediating stress signals between hepatic subcellular stress-responding machinery and in the onset of hepatic steatosis under conditions of metabolic stress.

Characterizing the landscape of cervical squamous cell carcinoma immune microenvironment by integrating the single-cell transcriptomics and RNA-Seq.

BACKGROUND: Cervical squamous cell carcinoma (CSCC), caused by the infection of high-risk human papillomavirus, is one of the most common malignancies in women worldwide. METHODS: RNA expression data, including those from the Cancer Genome Atlas, Gene Expression Omnibus, and Genotype-Tissue Expression databases, were used to identify the expression of RNAs in normal and tumor tissue. Correlation analysis was performed to identify the immune-related long noncoding RNAs (IRLs) and hypoxia-related genes (IRHs) that can influence the activity of the immune system. Prognosis models of immune-related RNAs (IRRs) were used to construct a coexpression network of the immune system. We identified the role of IRRs in immunotherapy by correlation analysis with immune checkpoint genes (ICGs). We then validated the expression data by integrating two single-cell sequencing data sets of CSCC to identify the key immune features. RESULTS: In total, six immune-related gene (IRG), four IRL, and five IRH signatures that can significantly influence the characteristics of the tumor immune microenvironment (TIME) were selected using machine learning methods. The expression level of ICGs was significantly upregulated in GZMB+ CD8+ T-cells and tumor-associated macrophages (TAMs) in tumor tissues. TGFBI+ TAMs are a kind of blood-derived monocyte-derived M0-like TAM linked to hypoxia and a poor prognosis. IFI30+ M1-like TAMs participate in the process of immune-regulation and showed a role in the promotion of CD8+ T-cells and Type 1 T helper (Th1)/Th2 cells in the coexpression network, together with several IRLs, IRGs, and ICGs. CONCLUSIONS: CD16+ monocyte-derived IFI30+ TAMs participated in our coexpression network to regulate the TIME, showing the potential to be a novel immunotherapy target. The enrichment of M0-like TAMs was associated with a worse prognosis in the high-risk score group with IRH signatures. Remarkably, M0-like TAMs in tumor tissues overexpressed TGFBI and were associated with several well-known tumor-proliferation pathways.

Activation of hepatic CREBH and Insig signaling in the anti-hypertriglyceridemic mechanism of R-α-lipoic acid.

The activation of sterol regulatory element binding proteins (SREBPs) is regulated by insulin-induced genes 1 and 2 (Insig-1 and Insig-2) and SCAP. We previously reported that feeding R-α-lipoic acid (LA) to Zucker diabetic fatty (ZDF) rats improves severe hypertriglyceridemia. In this study, we investigated the role of cyclic AMP-responsive element binding protein H (CREBH) in the lipid-lowering mechanism of LA and its involvement in the SREBP-1c and Insig pathway. Incubation of McA cells with LA (0.2 mM) or glucose (6 mM) stimulated activation of CREBH. LA treatment further induced mRNA expression of Insig-1 and Insig-2a, but not Insig-2b, in glucose-treated cells. In vivo, feeding LA to obesity-induced hyperlipidemic ZDF rats activated hepatic CREBH and stimulated transcription and translation of Insig-1 and Insig-2a. Activation of CREBH and Insigs induced by LA suppressed processing of SREBP-1c precursor into nuclear SREBP-1c, which subsequently inhibited expression of genes involved in fatty acid synthesis, including FASN, ACC and SCD-1, and reduced triglyceride (TG) contents in both glucose-treated cells and ZDF rat livers. Additionally, LA treatment also decreased abundances of very low density lipoprotein (VLDL)-associated apolipoproteins, apoB100 and apoE, in glucose-treated cells and livers of ZDF rats, leading to decreased secretion of VLDL and improvement of hypertriglyceridemia. This study unveils a novel molecular mechanism whereby LA lowers TG via activation of hepatic CREBH and increased expression of Insig-1 and Insig-2a to inhibit de novo lipogenesis and VLDL secretion. These findings provide novel insight into the therapeutic potential of LA as an anti-hypertriglyceridemia dietary molecule.