LC–MS-Based Metabolomics and Lipidomics Study of High-Density-Lipoprotein-Modulated Glucose Metabolism with an apoA‑I Knockout Mouse Model

Type 2 diabetes mellitus (T2DM) has become a tremendous problem in public health nowadays. High-density lipoprotein (HDL) refers to a group of heterogeneous particles that circulate in blood, and a recent research finds that HDL acts a pivotal part of glucose metabolism. To understand systemic metabolic changes correlated with HDL in glucose metabolism, we applied LC–MS-based metabolomics and lipidomics to detect metabolomic and lipidomic profiles of plasma from apoA-I knockout mice fed a high-fat diet. Multivariate analysis was applied to differentiate apoA-I knockout mice and controls, and potential biomarkers were found. Pathway analysis demonstrated that several metabolic pathways such as aminoacyl-tRNA biosynthesis, arginine and proline metabolism, and phenylalanine, tyrosine, and tryptophan biosynthesis were dysregulated in apoA-I knockout mice. This study may provide a new insight into the underlying pathogenesis in T2DM and prove that LC–MS-based metabolomics and lipidomics are powerful approaches in finding potential biomarkers and disturbed pathways

Construction of Nanodroplet/Adiposome and Artificial Lipid Droplets

The lipid droplet (LD) is a cellular organelle that consists of a neutral lipid core with a monolayer-phospholipid membrane and associated proteins. Recent LD studies demonstrate its importance in metabolic diseases and biofuel development. However, the mechanisms governing its formation and dynamics remain elusive. Therefore, we developed an <i>in vitro</i> system to facilitate the elucidation of these mechanisms. We generated sphere-shaped structures with a neutral lipid core and a monolayer-phospholipid membrane by mechanically mixing neutral lipids and phospholipids followed by a two-step purification. We named the nanodroplet “adiposome”. We then recruited LD structure-like/resident proteins to the adiposome, including the bacterial MLDS, Caenorhabditis elegans MDT-28/PLIN-1, or mammalian perilipin-2. In addition, adipose triglyceride lipase (ATGL) and apolipoprotein A1 (apo A-I) were recruited to adiposome. We termed the functional protein-coated adiposomes, Artificial Lipid Droplets (ALDs). With this experimental system, different proteins can be recruited to build ALDs for some biological goals and potential usage in drug delivery

Organic-Nanowire–SiO₂ Core–Shell Microlasers with Highly Polarized and Narrow Emissions for Biological Imaging

Development of luminescence probes with polarized and narrow emissions simultaneously is helpful for removing multiply scattered light and enables multiplexing detection, but it remains challenging to use conventional organic dyes, fluorescence proteins, and quantum dots. Here, we demonstrated smart one-dimensional microlaser probes (MLPs) by coating a thin layer of silica shell on the surface of organic nanowires (ONWs) of 1,4-dimethoxy-2,5-di­[4′-(methyl­thio)­styryl]­benzene (TDSB), namely, ONW@SiO₂ core–shell structures. Different from the Fabry–Pérot (FP) cavity formed between two end-faces of semiconductor nanowires, whispering gallery mode (WGM) microresonators are built within the rectangular cross section of ONW@SiO₂ MLPs. This enables a lasing threshold as low as 1.54 μJ/cm², above which lasing emissions are obtained with a full width at half-maximum (fwhm) < 5 nm and a degree of polarization (DOP) > 83%. Meanwhile, small dimensions of ONW@SiO₂ MLPs with a side-length of ca. 500 nm and a length of 3–8 μm help to reduce their perturbations in living cells. With the help of mesoporous silica shells, which provide both high biocompatibility and good photostability, ONW@SiO₂ MLPs can be easily introduced into the cell cytoplasm through natural endocytosis. Using their narrow and highly polarized lasing emissions in vitro, we demonstrate that it is possible to tag individual cells using ONW@SiO₂ MLPs with high stability

DataSheet2_Metabolic profiling reveals altered tryptophan metabolism in patients with kawasaki disease.CSV

Kawasaki disease (KD) is a childhood vasculitis disease that is difficult to diagnose, and there is an urgent need for the identification of accurate and specific biomarkers. Here, we aimed to investigate metabolic alterations in patients with KD to determine novel diagnostic and prognostic biomarkers for KD. To this end, we performed untargeted metabolomics and found that several metabolic pathways were significantly enriched, including amino acid, lipid, and tryptophan metabolism, the latter of which we focused on particularly. Tryptophan-targeted metabolomics was conducted to explore the role of tryptophan metabolism in KD. The results showed that Trp and indole acetic acid (IAA) levels markedly decreased, and that l-kynurenine (Kyn) and kynurenic acid (Kyna) levels were considerably higher in patients with KD than in healthy controls. Changes in Trp, IAA, Kyn, and Kyna levels in a KD coronary arteritis mouse model were consistent with those in patients with KD. We further analyzed public single-cell RNA sequencing data of patients with KD and revealed that their peripheral blood mononuclear cells showed Aryl hydrocarbon receptor expression that was remarkably higher than that of healthy children. These results suggest that the Trp metabolic pathway is significantly altered in KD and that metabolic indicators may serve as novel diagnostic and therapeutic biomarkers for KD.</p

DataSheet1_Metabolic profiling reveals altered tryptophan metabolism in patients with kawasaki disease.CSV

Kawasaki disease (KD) is a childhood vasculitis disease that is difficult to diagnose, and there is an urgent need for the identification of accurate and specific biomarkers. Here, we aimed to investigate metabolic alterations in patients with KD to determine novel diagnostic and prognostic biomarkers for KD. To this end, we performed untargeted metabolomics and found that several metabolic pathways were significantly enriched, including amino acid, lipid, and tryptophan metabolism, the latter of which we focused on particularly. Tryptophan-targeted metabolomics was conducted to explore the role of tryptophan metabolism in KD. The results showed that Trp and indole acetic acid (IAA) levels markedly decreased, and that l-kynurenine (Kyn) and kynurenic acid (Kyna) levels were considerably higher in patients with KD than in healthy controls. Changes in Trp, IAA, Kyn, and Kyna levels in a KD coronary arteritis mouse model were consistent with those in patients with KD. We further analyzed public single-cell RNA sequencing data of patients with KD and revealed that their peripheral blood mononuclear cells showed Aryl hydrocarbon receptor expression that was remarkably higher than that of healthy children. These results suggest that the Trp metabolic pathway is significantly altered in KD and that metabolic indicators may serve as novel diagnostic and therapeutic biomarkers for KD.</p

HepG2 k cells express elevated level of VEGFA.

<p>(A) Expression of p-Akt, HIF-1α and VEGFA in parental HepG2 and HepG2 k cells were detected by western blot analysis. (B) The mRNA expression of VEGFA in parental HepG2 and HepG2 k cells was assayed using real time PCR. *, <i>P</i><0.05. (C) VEGFA concentration in conditioned media from parental HepG2 and HepG2 k cells was measured by ELISA analysis. **, <i>P</i><0.01. (D) The expression of HIF-1α and VEGFA in parental HepG2 and HepG2 k cells treated with or without LY294002 (20 µM) for 24 h were analyzed by western blot analysis. (E) VEGFA concentration in conditioned media was detected by ELISA analysis. ***, <i>P</i><0.001 versus parental HepG2 cells control respectively; ^###, <i>P</i><0.001 versus HepG2 k cells; ns, no significance.</p

Additional file 2: of Sorafenib suppresses the epithelial-mesenchymal transition of hepatocellular carcinoma cells after insufficient radiofrequency ablation

The supplementary data. (DOCX 4641 kb

Additional file 1: of Sorafenib suppresses the epithelial-mesenchymal transition of hepatocellular carcinoma cells after insufficient radiofrequency ablation

NC3Rs ARRIVE Guidelines Checklist. (DOCX 664 kb

The viability of HepG2 cells and sublines derived from HepG2 cells after hyperthermia.

<p>(A) HepG2 cells were cultured after 47°C heat treatment. The 24 h, 48 h and 72 h cell viability of HepG2 cells with or without 47°C heat treatment were measured using MTT assay. (B) Twenty-four sublines were established after 47°C heat treatment for 10 min as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037266#s2" target="_blank">method</a>. The 24 h, 48 h, and 72 h viability was evaluated by MTT assay after 24 sublines were established. par, parental HepG2 cells; a–x, sublines derived from the HepG2 cells. (C) The 24 h, 48 h and 72 h viability of representative sublines of HepG2 cells were evaluated by MTT assay. (D) Parental HepG2 and HepG2 k cells were treated with 49°C or 50°C 10 min. The 4 h, 12 h, 24 h and 48 h cell viability were measured by MTT assay. *, <i>P</i><0.05; **, <i>P</i><0.01; ***, <i>P</i> <0.001. Data are the representative results of three independent experiments. The coefficients of variation (CV) of all assays were shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037266#pone.0037266.s001" target="_blank">Supporting Information S1</a>.</p