Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Assembly and characterization of human 3D liver spheroids via DNA origami NAC-linkers. (A) Schematic of 3D liver spheroid self-assembly from primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells using NAC-linkers. (B) Atomic force microscopy image of NAC-linkers. Scale bars, 200 nm. (C) 1% agarose gel electrophoresis confirming cholesterol-modified NAC-linkers assembly (lanes: DNA marker, M13mp18 scaffold, and NAC-linkers). (D) Bright-field image of a mature spheroid. (E) Hematoxylin and eosin (H&E) staining of a spheroid section. (F) Immunofluorescence staining of cell type markers in human 3D liver spheroids: albumin (ALB, hepatocytes), CD31 (endothelial cells), and CD68 (Kupffer cells). Scale bars, 200 μm.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Microscopy, Agarose Gel Electrophoresis, Modification, Marker, Staining, Immunofluorescence

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Adaptation and pathogenesis of MKWV in human 3D liver spheroids. (A) Schematic of serial passaging of the HLJ1 strain in spheroids, yielding the adapted NAC-Org5 strain. (B, C) Viral RNA copies (B) and TCID₅₀ titers (C) across passages (P1-P5). (D) Bright-field image of spheroids infected with passage 5 (P5) virus, showing structural disruption. Scale bar, 100 μm. (E) Quantification of spheroid diameter post-infection. (F) Transmission electron micrographs of virions within cytoplasmic vesicles of infected spheroids. Scale bars: 1 μm (left), 200 nm (right). (G) Representative images and quantification of nuclei showing infection-induced cell death. Scale bar, 200 μm. (H) Western blot detecting cleaved caspase-3 in spheroids at 48 and 72 h post-infection (hpi). (I) Multiplex immunofluorescence showing NAC-Org5 tropism for CD31 + endothelial cells and CD68 + Kupffer cells, with weaker detection in ALB + hepatocytes. Scale bar, 200 μm. (J) Functional assessment of infected spheroids: ATP (viability), ALT/AST/LDH (damage), ALB/urea (synthetic function). (K) RT-qPCR analysis of pro-inflammatory cytokine mRNA expression, normalized to β-actin. Data are mean ± SD ( n = 5 biological replicates). * p < 0.05, ** p < 0.01.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Passaging, Infection, Virus, Disruption, Transmission Assay, Western Blot, Multiplex Assay, Immunofluorescence, Functional Assay, Quantitative RT-PCR, Expressing

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Pathogenicity of the NAC-Org5 strain in murine models. (A) Experimental schematic for intracranial (3-day-old) and intraperitoneal (3-week-old) inoculation of BALB/c mice. (B, C) Survival (B) and weight change (C) of suckling mice after NAC-Org5 infection. (D) Viral load in tissues and blood of suckling mice at 7 dpi. (E, F) Survival (E) and weight change (F) of 3-week-old mice. (G) Viral load in tissues and blood of 3-week-old mice at 7 dpi. Data are from 3 independent experiments. (H) Representative H& E -stained liver sections from 3-week-old mice at 7 and 15 dpi, showing inflammatory infiltrates and hepatocyte necrosis that resolves by 15 dpi. Scale bar, 100 μm. *** p < 0.001.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Infection, Staining

Journal: Materials Today Bio

Article Title: Injectable chitosan-based hydrogel via in situ gelation modulates the inflammatory microenvironment and facilitates minimally invasive repair of peripheral nerve injury

doi: 10.1016/j.mtbio.2026.102814

Figure Lengend Snippet: Pro-angiogenic and pro-migratory effects of hydrogels. Immunofluorescence staining shows blood vessel formation of HUVECs in LPS-macrophage condition medium (A). Scratch assays and Transwell migration assays of HUVECs (B) and L929 cells (C). Quantification of junctions (D), branches (E), wound closure percentage (F–G) and migrated cells (H–I). One-way ANOVA with Tukey's post hoc test and n = 3 for D-I; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. ANOVA, analysis of variance; CS, chitosan; IBU, ibuprofen; GP, genipin; MA, methacrylic anhydride; LPS, lipopolysaccharides; HUVEC, human umbilical vein endothelial cell; n.s., not significant.

Article Snippet: Mouse fibroblasts (L929, ATCC), human umbilical vein endothelial cells (HUVECs, ATCC), and mouse macrophages (RAW 264.7, ATCC) were provided by the Cell Bank of the Chinese Academy of Sciences.

Techniques: Immunofluorescence, Staining, Migration

Journal: Regenerative Therapy

Article Title: Airway basal stem cell derived extracellular vesicles promote lung repair in chronic obstructive pulmonary disease

doi: 10.1016/j.reth.2026.101068

Figure Lengend Snippet: Characterization of BC-EVs. (a) Representative transmission electron microscopy image of BC-EVs. Scale bar, 100 nm. (b) Size distribution of BC-EVs determined by Nano-Flow Cytometry. (c) Western blotting analysis showing the expression of EV markers TSG101 and CD63, and the negative marker Calnexin. (d) Representative images of HUVECs tube formation after treatment with DMEM, BC-derived EVs, or MSC-derived EVs. Scale bar, 100 μm. (e) Quantification of the number of tubes formed in each group (n = 5 technical replicates). (f) Cell viability of BCs cultured under serum-depleted conditions and treated with DMEM, or increasing concentrations of BC-EVs or MSC-EVs (n = 3 technical replicates). (e) and (f) All data are presented as mean ± SEM. Statistical analysis was performed using a one-way ANOVA with Tukey's multiple comparisons test. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were obtained from the American Type Culture Collection (ATCC) and cultured in DMEM supplemented with 10 % fetal bovine serum.

Techniques: Transmission Assay, Electron Microscopy, Flow Cytometry, Western Blot, Expressing, Marker, Derivative Assay, Cell Culture

Journal: Molecular Medicine Reports

Article Title: PTUPB, a soluble epoxide hydrolase/cyclooxygenase-2 dual inhibitor, reduces endothelial-to-mesenchymal transition and improves doxorubicin-induced vascular and cardiac toxicity

doi: 10.3892/mmr.2026.13810

Figure Lengend Snippet: PTUPB decreases DOX-induced sEH and COX-2 expression in human endothelial cells. (A) Basal gene expression levels of sEH and COX-2 in endothelial cells. EA.hy926 cells were treated with 0.5 µM, 1 µM and 2 µM DOX for 24H. Gene expression levels of (B) sEH and (C) COX-2 in DOX-treated cells. Viability of EA.hy926 cells treated with either (D) PTUPB alone or (E) in combination with DOX. EA.hy926 cells were treated with 2 µM DOX with or without 1 µM PTUPB for 24 h. Gene expression levels of (F) sEH and (G) COX-2. (H) Activity level of sEH. One-way ANOVA followed by Tukey's multiple comparison tests was performed. + P<0.05 vs. control, *P<0.05 vs. DOX). sEH, soluble epoxide hydrolase; COX-2, cyclooxygenase-2; DOX, doxorubicin; PTUPB, 4-[5-phenyl-3-[3-[[[[4-(trifluoromethyl)phenyl]amino]carbonyl]amino]propyl]-1H-pyrazol-1-yl]-benzenesulfonamide; DHET, dihydroxyeicosatrienoic acid; Con, control.

Article Snippet: MDA-MB-231 cells (cat. no. HTB-26; American Type Culture Collection) and EA.hy926 human endothelial cells (cat. no. CRL-2922; American Type Culture Collection) were cultured in 75-cm 2 tissue-culture flasks that contained Dulbecco's Modified Eagle Medium F-12 Nutrient Mixture supplemented with 10% w/v fetal bovine serum and 1% w/v Antibiotic-Antimycotic (all Gibco; Thermo Fisher Scientific, Inc.) in a humid environment at 37°C with 5% CO 2 .

Techniques: Expressing, Gene Expression, Activity Assay, Comparison, Control

Journal: Molecular Medicine Reports

Article Title: PTUPB, a soluble epoxide hydrolase/cyclooxygenase-2 dual inhibitor, reduces endothelial-to-mesenchymal transition and improves doxorubicin-induced vascular and cardiac toxicity

doi: 10.3892/mmr.2026.13810

Figure Lengend Snippet: Inhibition of soluble epoxide hydrolase and cyclooxygenase-2 using PTUPB attenuates DOX-induced endothelial-to-mesenchymal transition. EA.hy926 cells were treated with 2 µM DOX with or without 1 µM PTUPB for 24 h. Gene expression levels of mesenchymal markers (A) ASMA, (B) SMA22, (C) VIM, (D) CDH2, (E) TGF-β, (F) SNAI1 and (G) SNAI2, and (H) endothelial marker CD31. (I) Ratio of mesenchymal to endothelial cells in cells treated with DOX with or without 1 µM PTUPB was measured based on morphological images. (J) Representative morphological images of EA.hy926 cells incubated with DOX with or without 1 µM PTUPB for 24 h at a magnification of ×10 (scale bar, 100 µm). The light red arrow indicates mesenchymal cells (long, spindle-like cells). The dark red arrow indicates endothelial cells (cobblestone monolayer-like cells). One-way ANOVA followed by Tukey's multiple comparison test was performed to determine the significant differences between groups ( + P<0.05 vs. control, *P<0.05 vs. DOX). PTUPB, 4-[5-phenyl-3-[3-[[[[4-(trifluoromethyl)phenyl]amino]carbonyl]amino]propyl]-1H-pyrazol-1-yl]-benzenesulfonamide; DOX, doxorubicin; ASMA, smooth muscle actin α2; SMA22, smooth muscle protein 22α; VIM, vimentin; CDH2, cadherin-2; SNAI1, snail family transcriptional repressor 1; SNAI2, snail family transcriptional repressor 2.

Article Snippet: MDA-MB-231 cells (cat. no. HTB-26; American Type Culture Collection) and EA.hy926 human endothelial cells (cat. no. CRL-2922; American Type Culture Collection) were cultured in 75-cm 2 tissue-culture flasks that contained Dulbecco's Modified Eagle Medium F-12 Nutrient Mixture supplemented with 10% w/v fetal bovine serum and 1% w/v Antibiotic-Antimycotic (all Gibco; Thermo Fisher Scientific, Inc.) in a humid environment at 37°C with 5% CO 2 .

Techniques: Inhibition, Gene Expression, Marker, Incubation, Comparison, Control