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

Additional file 1: of 15-oxoeicosatetraenoic acid mediates monocyte adhesion to endothelial cell

The details of patients are provided as follow. (DOCX 32 kb

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

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

YC-1 and VEGFA siRNA inhibit HepG2 cell viability.

<p>The parental HepG2 and HepG2 k cells were treated with or without YC-1(5 µM) or transfected with or without VEGFA siRNA. (A) The expression of HIF-1α and VEGFA was detected by western blot analysis. (B) VEGFA concentration in conditioned media was detected by ELISA analysis. **, <i>P</i><0.01, ***, <i>P</i><0.001 versus parental HepG2 cells control respectively; ^###, <i>P</i><0.001 versus HepG2 k cells. (C) Parental HepG2 and HepG2 k cells were transfected with or without VEGFA siRNA for 24 h, 48 h, and 72 h, cell viability was measured by MTT assay. ***, <i>P</i><0.01; ns, no significance. (D) Parental HepG2 and HepG2 k cells were treated with or without YC-1 (5 µM) for 24 h, 48 h, and 72 h, cell viability was measured by MTT assay. ***, <i>P</i><0.001; ns, no significance.</p

The enhanced pro-angiogenic ability of HepG2 k is in a HIF-1α/VEGFA dependent manner <i>in vitro</i>.

<p>(A) HUVECs were treated with conditioned media from parental HepG2 or HepG2 k cells with addition of bevacizumab (0.5 mg/ml) or control IgG for 24 h. Cell viability were quantified by MTT assay. (B–C) HUVEC migration <i>in vitro</i> in response to conditioned media from HepG2 k or parental HepG2 cells with addition of bevacizumab (0.5 mg/ml) or control IgG was assayed after 12 h. The number of migrated cells was quantified by counting 10 random fields at ×100 magnification. (D–E) HUVEC tube formation in response to conditioned media from HepG2 k or parental HepG2 cells with addition of bevacizumab (0.5 mg/ml) or control IgG after 20 h was assayed. The length of tube was evaluated by counting 10 random fields at ×100 magnification. (F–H) The parental HepG2 and HepG2 k cells were treated with or without VEGFA siRNA or 5 µM YC-1, and the conditioned media was collected. The effect of various conditioned media on HUVEC 24 h viability was quantified by MTT assay. The effect of various conditioned media on HUVEC migration for 12 h was assayed. The number of migrated cells was quantified by counting 10 random fields at ×100 magnification. The effect of various conditioned media on HUVEC tube formation for 20 h was assayed. The tube length was evaluated by counting 10 random fields at ×100 magnification. *, <i>P</i><0.05, ***, <i>P</i><0.001 versus parental HepG2 cell control; ^##, <i>P</i><0.01, ^###, <i>P</i><0.001 versus HepG2 k cells control; ns, no significance.</p

Bevacizumab impaired the tumor growth and angiogenesis in tumor-bearing mice <i>in vivo</i>.

<p>2×10⁶ HepG2 k or parental HepG2 cells in cells in 200 µl phosphate buffered saline were injected by subcutaneous injection to obtain s. c. tumors. (A) Average tumor volume is shown for the HepG2 tumors. Parental HepG2 tumor with control IgG (n = 5), Parental HepG2 tumor with Bevacizumab (n = 5), HepG2 k tumor with control IgG (n = 5) and HepG2 k tumor with Bevacizumab (n = 5). **, <i>P</i><0.01. (B) Mice were killed after 28 days implantation and the tumor tissues were removed and weighed. *, <i>P</i><0.05. (C–D) Twenty-eight days after implantation, the numbers of new microvessels marked with CD34 (arrow heads) in the subcutaneous tumors were quantified by performing new vessel counts of 10 random fields at ×400 magnification. ^*, <i>P</i><0.05, ^**, <i>P</i><0.01 versus parental HepG2 tumor respectively; ^###, <i>P</i><0.001 versus HepG2 k tumor. (E) Immunohistochemistry analysis of the expression of VEGF in implanted tumors.</p

Additional file 1: of Myeloperoxidase-oxidized high density lipoprotein impairs atherosclerotic plaque stability by inhibiting smooth muscle cell migration

Supplemental methods and supplemental results. (DOC 125 kb

sj-docx-1-tar-10.1177_17534666231224692 – Supplemental material for The significance of dynamic monitoring plasma TMAO level in pulmonary arterial hypertension – a cohort study

Supplemental material, sj-docx-1-tar-10.1177_17534666231224692 for The significance of dynamic monitoring plasma TMAO level in pulmonary arterial hypertension – a cohort study by Yicheng Yang, Xin Li, Peizhi Wang, Songren Shu, Bingyang Liu, Yanru Liang, Beilan Yang, Zhihui Zhao, Qin Luo, Zhihong Liu, Lemin Zheng, Qixian Zeng and Changming Xiong in Therapeutic Advances in Respiratory Disease</p