Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Simultaneous application of IL-4 and Arg promotes efferocytosis and improves the pro-regenerative microenvironment (A) Schematic illustration of the in vitro efferocytosis assessment. (B and C) Macrophage efferocytosis assessment via flow cytometry, showing representative scatterplots of macrophages engulfing apoptotic cells (B) and quantification of the efferocytosis rate after 24 h (C). (D) Morphological evaluation of macrophage efferocytosis, showing representative cell tracking images of macrophages (labeled with CMFDA) engulfing apoptotic cells (labeled with CM-Dil) after 24 h of co-culture. (E) Quantification of the efferocytosis rate after 6, 24, and 48 h determined by flow cytometry under Control and IL-4 + Arg conditions. (F) Experimental design for detecting microenvironmental changes following efferocytosis. (G) ROS quantification following efferocytosis. (H) Enzyme-linked immunosorbent assay (ELISA) quantification of TGF-β1 and IL-10 in the culture supernatants. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (C, G, and H) or two-way ANOVA followed by Sidak’s multiple comparisons test (E). Sample sizes: n = 3 biological replicates (B–E); n = 5 biological replicates, each performed with 3 technical replicates (G) and n = 3 biological replicates, each performed with 3 technical replicates (H). Scale bars, 100 μm (D) and 20 μm (inset).

Article Snippet: The secretion levels of anti-inflammatory cytokines were quantified using a Mouse TGF-β1 ELISA Kit (ABclonal, Wuhan, China) and a Mouse IL-10 ELISA Kit (Elabscience, Wuhan, China).

Techniques: In Vitro, Flow Cytometry, Cell Tracking Assay, Labeling, Co-culture Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Transcriptomic changes underlying synergistic regulation of macrophages by IL-4 and Arg (A) RT-qPCR analysis of M2-related marker gene expression ( Tgfb1 , Il10 , Arg1 , and Cd206 ) in macrophages. (B) Schematic illustration of the bone marrow-derived macrophages (BMDMs) isolation, total RNA extraction, and transcriptomic sequencing workflow. (C) Functional annotation of upregulated genes using Gene Ontology (GO) enrichment analysis. (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis emphasizing significant signaling pathways. (E) RT-qPCR analysis of Tgfb1 and Il10 mRNA expression levels of macrophage after treatment with the IL-4 + Arg combination in the presence or absence of nor-NOHA. (F) ELISA quantification of secreted TGF-β1 and IL-10 protein concentrations in the culture supernatants under nor-NOHA inhibition. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (A, E, and F). Sample sizes: n = 3 biological replicates (C and D); n = 3 (A and F) and 4 (E) biological replicates, each performed with 3 technical replicates.

Article Snippet: The secretion levels of anti-inflammatory cytokines were quantified using a Mouse TGF-β1 ELISA Kit (ABclonal, Wuhan, China) and a Mouse IL-10 ELISA Kit (Elabscience, Wuhan, China).

Techniques: Quantitative RT-PCR, Analysis, Marker, Gene Expression, Derivative Assay, Isolation, RNA Extraction, Sequencing, Functional Assay, Protein-Protein interactions, Expressing, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet:

Article Snippet: The secretion levels of anti-inflammatory cytokines were quantified using a Mouse TGF-β1 ELISA Kit (ABclonal, Wuhan, China) and a Mouse IL-10 ELISA Kit (Elabscience, Wuhan, China).

Techniques: Cell-Signaling, Recombinant, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Software

Journal: Cell Biochemistry and Function

Article Title: Capybara Oil‐Based Nanoemulsion Attenuates Obesity‐Induced Renal Injury Through Modulation of Inflammation, Oxidative Stress, and Fibrosis in Mice

doi: 10.1002/cbf.70316

Figure Lengend Snippet: Immunostaining and quantification of TNF‐α and IL‐10 cytokines in the renal cortex of the experimental groups. (A) Representative photomicrographs of TNF‐α immunostaining. (B) Representative photomicrographs of IL‐10 immunostaining. Arrows indicate positive immunostaining. (C) Represents quantification of TNF‐α‐positive immunostaining and (D) Represents quantification of IL‐10‐positive immunostaining, expressed as pixels/µm 2 . Scale bars: 50 µm (A) and 200 µm (B). Data are expressed as mean ± SEM ( n = 6 per group). (a) p < 0.05 versus CD; (b) p < 0.05 versus CD + NE‐CO; (c) p < 0.05 versus CD + CO; (d) p < 0.05 versus CD + NE‐W; (e) p < 0.05 versus HFD; and (f) p < 0.05 versus HFD + CO.

Article Snippet: We used anti‐TNF‐α (sc‐52746), anti‐TGF‐β (sc‐130348), IL‐10 (sc‐8438), and anti‐α‐SMA (sc‐53142) (dilution 1:200, Santa Cruz Biotechnology, Dallas, Texas, USA) antibodies.

Techniques: Immunostaining

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Systemic TREM1 ablation attenuates neuroinflammation and neurological deficits after TBI. (a) Schematic timeline of the experimental design. WT and Trem1 −/− mice were subjected to CCI or sham surgery, followed by longitudinal behavioral and histological assessments. (b, c) Analysis of the neuroinflammatory profile in the perilesional cortex at 3 days post‐TBI. (b) Representative images of inflammatory cytokines in TBI lesions detected by the Proteome Profiler Array Mouse Cytokine Array Panel A in WT and Trem1 −/− mice at 3 days post‐TBI. (c) Quantification of inflammatory cytokines in TBI lesions of WT and Trem1 −/− mice at 3 days post‐TBI or sham treatment by ELISA ( n = 5 mice per group, two‐way ANOVA followed by Tukey's multiple‐comparison test). (d) Representative immunofluorescence images of macrophages (IBA1 + , green) and the reactivation marker CD86 (red) in the TBI cortex. Trem1 ablation significantly reduced reactive polarization (scale bar, 50 µm). (e) Evaluation of neuronal apoptosis. Representative co‐staining of TUNEL (green) and NeuN (red) (left) ( n = 3 mice per group, scale bar, 100 µm). Yellow arrows indicate apoptotic neurons. (f) Assessment of white matter integrity. Representative images of Myelin Basic Protein (MBP, green) and NeuN (red) (left) and quantification of MBP fluorescence intensity (right) (scale bar, 100 µm). (g) Representative immunofluorescence images of hippocampal neurons (NeuN, red) showing preserved cellular density in Trem1 −/− mice ( n = 3 mice per group; scale bar, 100 µm). (h) Brain water content analysis at 1day and 3 d post‐TBI, indicating alleviated cerebral edema in Trem1 −/− mice ( n = 6 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (i, j) Assessment of blood‐brain barrier (BBB) permeability. (i) Representative images of Evans Blue extravasation. (j) Quantification of dye leakage ( n = 6 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (k, l) Molecular validation of tissue preservation. (k) Representative Western blots of the neuronal marker beta3‐tubulin, synaptic marker Synaptophysin (Syn), and tight junction protein Occludin. l, Densitometric quantification normalized to GAPDH ( n = 3 independent experimental replicates; two‐way ANOVA followed by Tukey's multiple‐comparison test). (m–o) Assessment of sensorimotor function. m, Latency to fall in the accelerating Rotarod test ( n = 8 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (n) Performance in the fixed‐speed (30 rpm) Rotarod test ( n = 8 mice per group, one‐way ANOVA followed by Tukey's multiple‐comparison test). o, Modified Neurological Severity Score (mNSS) trajectory ( n = 8 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (p, q) Evaluation of anxiety‐like behavior and exploratory activity in the Open Field Test (OFT). (p) Representative locomotor tracking plots. (q) Quantification of the percentage of time spent in the center zone ( n = 8 per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). Data are presented as mean ± SD. Each dot represents an individual mouse, except in l, where each dot represents an independent experimental replicate. Statistical tests are specified for each panel above. Exact p values are provided in the Source Data. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐10 ELISA kit , ABclonal , # RK00016.

Techniques: Analysis, Enzyme-linked Immunosorbent Assay, Comparison, Immunofluorescence, Marker, Staining, TUNEL Assay, Fluorescence, Permeability, Biomarker Discovery, Tissue, Preserving, Western Blot, Modification, Activity Assay

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Generative AI‐driven design and mechanistic validation of a TREM1‐targeting LYTAC degrader. (a) Schematic of the RFdiffusion‐guided pipeline for de novo TREM1‐binding peptide design. Peptide backbones were generated around the Ig‐like domain of TREM1 and subsequently screened by AlphaFold3, ZDOCK, HDOCK, and Rosetta‐based energetic analysis to identify high‐ranking candidates. (b) Predicted structural models of Pep15, Pep17, and Pep457 in complex with TREM1, showing the predicted binding interfaces. (c) Temperature‐related intensity change (TRIC) assays showing concentration‐dependent target engagement of Pep15, Pep17, and Pep457 with TREM1. Left, schematic illustration of the TRIC assay principle; right, dose‐response curves of the indicated peptides ( n = 3 independent experiments). (d) ELISA‐based binding analysis of Pep15, Pep17, and Pep457 to immobilized TREM1. Top, schematic of the ELISA workflow; bottom, quantitative binding curves ( n = 3 independent experiments). (e) Surface plasmon resonance (SPR) analysis of Pep457 binding to TREM1. Top, schematic of the SPR assay; bottom, representative sensorgrams obtained at the indicated Pep457 concentrations. (f) Structural model of the bifunctional TREM1‐LYTAC, in which Pep457 engages TREM1 and the mannose‐6‐phosphate (M6P) moiety recruits the cation‐independent mannose‐6‐phosphate receptor (CI‐M6PR). (g) Representative confocal images showing partial colocalization of TREM1‐LYTAC with lysosomes following cellular uptake. TREM1‐LYTAC, green; lysosomes, red; nuclei, blue. Arrows indicate representative colocalized puncta. Scale bars as indicated. (h) ELISA quantification of cellular TREM1 protein following treatment with vehicle, scrambled‐LYTAC, TREM1‐LYTAC, or TREM1‐LYTAC in the presence of bafilomycin A1 (BafA1) or MG132 ( n = 5 independent experiments). (i) Representative flow‐cytometric histograms showing cell‐surface TREM1 expression after the indicated treatments. (j) Trem1 mRNA expression measured by qPCR after the indicated treatments ( n = 3 independent experiments; one‐way ANOVA followed by Tukey's multiple‐comparison test). (k) Requirement of both functional modules for TREM1 reduction. Cellular TREM1 protein levels were quantified after treatment with Pep457 plus M6P, Pep457 alone, M6P alone, TREM1‐LYTAC in the presence of CI‐M6PR blockade, or conjugated TREM1‐LYTAC ( n = 5 independent experiments; one‐way ANOVA followed by Tukey's multiple‐comparison test). (l) Plasma pharmacokinetic profile of free TREM1‐LYTAC after intravenous administration. Fluorescence intensity was fitted to a monoexponential decay model, yielding an apparent half‐life of 1.13 h ( n = 3 mice). (m) Proposed mechanism of TREM1‐LYTAC‐mediated degradation. TREM1‐LYTAC bridges cell‐surface TREM1 to CI‐M6PR, thereby promoting receptor internalization and lysosomal degradation. Data are mean ± SD. Each dot represents an independent experimental replicate or an individual animal. Statistical tests are specified for each quantitative panel above. Exact p values are provided in the Source Data. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐10 ELISA kit , ABclonal , # RK00016.

Techniques: Biomarker Discovery, Binding Assay, Generated, Analysis, Concentration Assay, Drug discovery, Enzyme-linked Immunosorbent Assay, SPR Assay, Confocal, Expressing, Comparison, Functional Assay, Clinical Proteomics, Fluorescence

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Engineering and validation of a BBB‐targeted, pH‐responsive nanodevice for spatiotemporal delivery of TREM1‐LYTAC. (a) Schematic illustration of APTL‐NP fabrication. TREM1‐LYTAC was conjugated to a generation‐4 PAMAM dendrimer through a terephthalaldehyde‐derived acid‐labile linker, followed by surface functionalization with Angiopep‐2 and PS‐PEG‐NHS to enable LRP1‐directed BBB targeting and improve systemic stability. (b) Representative transmission electron microscopy images of PAMAM‐based precursor particles and APTL‐NP (Scale bars, 1 µm). (c) Dynamic light scattering profiles showing the apparent hydrodynamic size distributions of G4 PAMAM and APTL‐NP under the indicated measurement conditions. (d) Time‐dependent changes in hydrodynamic diameter and polydispersity index of APTL‐NP incubated in PBS at pH 7.4 and 37°C. (e) pH‐dependent zeta‐potential profile of APTL‐NP. (f) Nanoparticle tracking analysis showing pH‐dependent particle size and concentration distributions after incubation at pH 7.5, 7.0, or 6.5. (g) pH‐responsive release of TREM1‐binding‐competent TREM1‐LYTAC from APTL‐NP. Released free TREM1‐LYTAC was separated by ultrafiltration and quantified using a TREM1‐binding ELISA with free TREM1‐LYTAC standards ( n = 3 independent experiments). (h) Representative EdU staining images of BMECs, HT22 cells, and NHAs treated with vehicle or APTL‐NP. (i) Quantification of EdU‐positive cells in the indicated cell types( n = 6 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (j) Representative flow‐cytometric plots for apoptosis analysis in BMECs, HT22 cells, and NHAs after treatment with vehicle or APTL‐NP. (k) Quantification of apoptotic cells in the indicated cell types( n = 6 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (l) Cell viability after 24 h or 48 h exposure to the indicated concentrations of APTL‐NP ( n = 3 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (m) Plasma pharmacokinetic profile of APTL‐NP after intravenous administration. The fluorescence signal was fitted to a monoexponential decay model, yielding an apparent half‐life of 8.38 h ( n = 3 mice). (n) Representative photographs and quantification of hemolysis induced by the indicated concentrations of APTL‐NP ( n = 3 independent experiments, one‐way ANOVA followed by Tukey's multiple‐comparison test). (o, p) Plasma C3a and C5a concentrations at the indicated time points after administration of vehicle or APTL‐NP ( n = 6 mice per group, Two‐way ANOVA followed by Tukey's multiple‐comparison test). (q) Representative H&E‐stained sections of the spleen, heart, lung, liver, and kidney from vehicle‐ and APTL‐NP‐treated mice (Scale bar, 100 µm). Data are presented as mean ± SD where applicable. For in vitro assays, n denotes independent experimental replicates; for in vivo analyses, n denotes individual mice. Statistical tests are specified for each quantitative panel above. Exact p values are provided in the Source Data. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐10 ELISA kit , ABclonal , # RK00016.

Techniques: Biomarker Discovery, Derivative Assay, Stability, Transmission Assay, Electron Microscopy, Dynamic Light Scattering Assay, Incubation, Zeta Potential Analyzer, Analysis, Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Staining, Comparison, Clinical Proteomics, Fluorescence, In Vitro, In Vivo

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Key resources table.

Article Snippet: IL‐10 ELISA kit , ABclonal , # RK00016.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software