Reactive Template and Confined Self-Activation Strategy: Three-Dimensional Interconnected Hierarchically Porous N/O-Doped Carbon Foam for Enhanced Supercapacitors

Herein, we report a new reactive template and confined self-activation strategy for the preparation of three-dimensional interconnected hierarchically porous N/O-doped carbon foam (3DHPNCF), which contains abundant micropores, mesopores, and macropores via using mesoporous silica SBA-15 as a hard template and ethylenediaminetetraacetic acid tripotassium (EDTA-3K) as a single carbon/nitrogen precursor, respectively. Importantly, potassium carbonate produced by the pyrolysis of EDTA-3K can effectively react with the SBA-15 silica skeleton to form water-soluble potassium silicate, after hot-water removal, leading to a continuous macropore structure. Simultaneously, a confined alkali activation reaction is carried out between potassium carbonate and in situ as-formed carbon to create meso/micropores. 3DHPNCF exhibits the high specific surface area and pore volume, as well as rich content of in situ-doped oxygen/nitrogen element, contributing to the high specific capacitance, good rate capability, and outstanding cycling stability. We provide a novel, green, and universal strategy for the design and fabrication of novel hierarchically porous carbons

A Novel Poly(ionic liquid) Interface-Free Two-Dimensional Monolithic Material for the Separation of Multiple Types of Glycoproteins

Currently, many types of affinity materials have been developed for the enrichment of glycoproteins potentially considered to be clinical biomarkers; however, they can not effectively distinguish between different glycoproteins and thus lack the functionality that may be the key to the diagnosis of specific diseases. In the present work, a novel interface-free 2D monolithic material has been developed for the separation of multiple types of glycoproteins, in which boronate-functionalized graphene acts as preconcentration segment and poly­(guanidinium ionic liquid) acts as separation segment. The resultant 2D material was characterized by X-ray photoelectron spectroscopy, elemental analysis, and electroosmotic flow analysis to demonstrate successful modification at each step. The performance of this 2D material was evaluated by capillary electrochromatography and allowed the successful online concentration and separation of five standard glycoproteins. The high separation efficiency can be largely attributed to the good orthogonality of boronate-functionalized graphene monolith and poly­(guanidinium ionic liquid) monolith

qRT-PCR validation of five genes (COL4A2, BMF, DUSP1, FOXA1and MLPH) in seven breast cancer cell lines (MDA-MB-231, MDA-MB-435, MDA-MB-468, MDA-MB-453, MCF-7, BT-474 and SK-BR-3).

<p>qRT-PCR validation of five genes (COL4A2, BMF, DUSP1, FOXA1and MLPH) in seven breast cancer cell lines (MDA-MB-231, MDA-MB-435, MDA-MB-468, MDA-MB-453, MCF-7, BT-474 and SK-BR-3).</p

Molecular Features of Triple Negative Breast Cancer: Microarray Evidence and Further Integrated Analysis

<div><p>Purpose</p><p>Breast cancer is a heterogeneous disease usually including four molecular subtypes such as luminal A, luminal B, HER2-enriched, and triple-negative breast cancer (TNBC). TNBC is more aggressive than other breast cancer subtypes. Despite major advances in ER-positive or HER2-amplified breast cancer, there is no targeted agent currently available for TNBC, so it is urgent to identify new potential therapeutic targets for TNBC.</p><p>Methods</p><p>We first used microarray analysis to compare gene expression profiling between TNBC and non-TNBC. Furthermore an integrated analysis was conducted based on our own and published data, leading to more robust, reproducible and accurate predictions. Additionally, we performed qRT-PCR in breast cancer cell lines to verify the findings in integrated analysis.</p><p>Results</p><p>After searching Gene Expression Omnibus database (GEO), two microarray studies were obtained according to the inclusion criteria. The integrated analysis was conducted, including 30 samples of TNBC and 77 samples of non-TNBC. 556 genes were found to be consistently differentially expressed (344 up-regulated genes and 212 down-regulated genes in TNBC). Functional annotation for these differentially expressed genes (DEGs) showed that the most significantly enriched Gene Ontology (GO) term for molecular functions was protein binding (GO: 0005515, P = 6.09E-21), while that for biological processes was signal transduction (GO: 0007165, P = 9.46E-08), and that for cellular component was cytoplasm (GO: 0005737, P = 2.09E-21). The most significant pathway was Pathways in cancer (P = 6.54E-05) based on Kyoto Encyclopedia of Genes and Genomes (KEGG). DUSP1 (Degree = 21), MYEOV2 (Degree = 15) and UQCRQ (Degree = 14) were identified as the significant hub proteins in the protein-protein interaction (PPI) network. Five genes were selected to perform qRT-PCR in seven breast cancer cell lines, and qRT-PCR results showed that the expression pattern of selected genes in TNBC lines and non-TNBC lines was nearly consistent with that in the integrated analysis.</p><p>Conclusion</p><p>This study may help to understand the pathogenesis of different breast cancer subtypes, contributing to the successful identification of therapeutic targets for TNBC.</p></div

The top 15 enriched GO terms of differentially expressed genes.

<p>A. molecular functions for DEGs (p value ≤ 9.35E-06); B. biological process for DEGs (p value ≤ 1.92E-07); C. cellular component for DEGs (p value ≤ 2.98E-09).</p

NET formation induced by complement-mediated phagocytosis against the invasion of <i>C. albicans</i>.

<p>(A) Serum-treated or non-treated <i>C. albicans</i> were added to neutrophil-like differentiated HL60 cells (the ratio of <i>C. albicans</i> to cell; 5∶1) in the presence of Sytox Green for 3 h at 37°C. Left panel shows representative fluorescence images and right panel shows frequency of Sytox green-positeve cells. Green shows Sytox Green staining. (B) Serum-treated <i>C. albicans</i> were added to neutrophil-like differentiated control cells and Rab27a-knockdown cells in the presence of both Hoechst 33342 and Sytox Green for 3 h (The ratio of <i>C. albicans</i> to cell; 5∶1). Left panel shows representative fluorescence images. Blue shows Hoechst 33342 staining and Green Sytox Green. Right panel shows the percentage of the cells in the individual stage (black: Hoechst 33342–positive; white: Sytox Green-positive and decondensed; shaded area: Sytox Green-positive and cloud-like shape). (C) Serum-treated <i>C. albicans</i> were added to neutrophil-like differentiated control cells and Rab27a-knockdown cells in the presence or absence of DPI for 3 h. Frequency of Sytox green-positive cells are shown. In (A) and (B), Scale bars indicate 10 µm. In (A), (B) and (C), the results of Rab27aKD clone 1 and Control clone 1 are shown and the data are the mean with SD from three independent experiments. Asterisks (*) mean that the difference is statistically significant (p values <0.01).</p

The effects of Rab27a-knockdown on PMA-induced NET formation.

<p>(A–C) Neutrophil-like differentiated control HL60 cells (Control; clone 1 and clone 2) and Rab27a-knockdown cells (Rab27aKD; clone 1 and clone 2) were stimulated with PMA. (A) Both cells fixed with PFA were stained with Hoechst 33342 (left). Cell lysates were assesed by immunoblotting analysis using specific antibody against H4cit3 and histone H4 (middle). PFA-fixed cells were stained with antibody against H4cit3 in the presence of Hoechst 33342 (right). Scale bars indicate 10 µm. (B) Representative fluorescence images at 4 hours after PMA treatment in the presence of both Hoechst 33342 and Sytox Green. Scale bars indicate 10 µm. Blue and green colors show Hoechst 33342-positive and Sytox Green-positive, respectively (left). The results of Rab27aKD clone 1 and Control clone 1 are shown as representative data. Frequency of Sytox Green-positive cells at indicated time points after PMA treatment (middle). The percentage of the cells in the individual stages at 4 h after PMA treatment (right). (C) The mean fluorescence intensity (MFI) after PMA treatment in the presence of APF at 30 min (left) or CM-H₂DCFDA at 20min (right). The mean value of MFI of control cells was adjusted to 1. In (B) and (C), the data are the mean with SD from three independent experiments.</p

Top 15 enriched KEGG pathway of DEGs.

<p>Top 15 enriched KEGG pathway of DEGs.</p

Process of NET formation after PMA treatment both in primary neutrophils and neutrophil-like differentiated HL60 cells.

<p>(A–C) Primary human neutrophils and neutrophil-like differentiated HL60 cells were stimulated with PMA. (A) NET formation was observed both in human neutrophils (a–c) and neutrophil-like differentiated HL60 cells (d–g). (a and d) Scanning electron microscopy (SEM) images of both cells indicating NET structures. Scale bars indicate 10 µm. (b and e) PFA-fixed cells were stained with antibody against histone H3 in the presence of Hoechst 33342. Scale bars indicate 20 µm. Extracellular histone H3 content in DNase I-treated supernatants were assessed by immunoblotting analysis in the time course study after PMA treatment. (c) PFA-fixed cells were stained with antibody against Rab27a in the presence of Hoechst 33342. (f) PFA-fixed cells were stained with antibody against H4cit3 in the presence of Hoechst 33342. Scale bars indicate 10 µm. (g) The amounts of histone H4cit3 and histone H4 in the cells in the time course study after PMA treatment were assesed by immunoblotting analysis using specific antibodies.(B) Both primary neutrophils and neutrophil-like differentiated HL60 cells were stimulated with PMA with a mixture of Hoechst 33342 and Sytox Green. Representative fluorescence images at indicated time points are shown. In the images of human neutrophils (right), x-y-z section images are also shown to indicate the spherical forms of the nuclei. Scale bars indicate 10 µm. Blue and green colors show Hoechst 33342-positive and Sytox Green-positive, respectively. (C) Cell stages in NET formation were classified into four groups by the degree of nuclei expansion and types of reagent staining (stage1–4). Nuclei are stained with Hoechst 33342 and showed lobulated shape and decondensation does not occur judging from z-section scanning (stage1), nuclei are stained with Hoechst 33342 and chromatin decondensation occurs like a spherical form (stage2), chromatin stained with Sytox Green like a spherical form (stage3), chromatin stained with Sytox Green shows a cloud-like spread form (stage4). The histogram shows the percentage of the cells in the individual stage at indicated time points. The data are the mean with SD from three independent experiments. Scale bars indicate 5 µm.</p

Heat-map image representing 66 genes that were significantly up-regulated or down-regulated (p<0.003) in TNBC compared with other breast cancer subtypes including LuminalA, LuminalB and HER2+.

<p>Heat-map image representing 66 genes that were significantly up-regulated or down-regulated (p<0.003) in TNBC compared with other breast cancer subtypes including LuminalA, LuminalB and HER2+.</p