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: Oncology Letters

Article Title: Cytotoxic, apoptotic and genotoxic effects of thymoquinone-oxime derivative on gastric cancer cells: An in vitro study

doi: 10.3892/ol.2026.15462

Figure Lengend Snippet: Effect of TQ-ox on apoptosis in HGEpiCs and AGS cells. Statistical significance was analyzed using one-way analysis of variance with Tukey's post hoc test. **P<0.01 and ***P<0.001 vs. control. Representative fluorescence microscopy images of acridine orange/ethidium bromide-stained cells showing live (green) and apoptotic (red/orange) cells in the control and 40 µM TQ-ox treated AGS groups. A total of ~50 cells were analyzed per condition. Scale bar, 100 µm. HGEpiC, human gastric epithelial cell; TQ-ox, thymoquinone-oxime.

Article Snippet: Primary HGEpiCs were obtained from Innoprot (cat. no. P10778).

Techniques: Control, Fluorescence, Microscopy, Staining

Journal: Oncology Letters

Article Title: Cytotoxic, apoptotic and genotoxic effects of thymoquinone-oxime derivative on gastric cancer cells: An in vitro study

doi: 10.3892/ol.2026.15462

Figure Lengend Snippet: Effect of TQ-ox on DNA damage in HGEpiCs and AGS cells following 24 h of TQ-ox treatment (5–40 µM). Statistical significance was analyzed using one-way analysis of variance with Tukey's post hoc test. **P<0.01 and ***P<0.001 vs. control. Representative fluorescence microscopy images showing nuclei (control) and comet tails (40 µM TQ-ox-treated cells), indicating fragmented DNA. A total of ~50 cells were analyzed per condition. Scale bar, 200 µm. HGEpiC, human gastric epithelial cell; TQ-ox, thymoquinone-oxime.

Article Snippet: Primary HGEpiCs were obtained from Innoprot (cat. no. P10778).

Techniques: Control, Fluorescence, Microscopy

Journal: Clinical and Translational Radiation Oncology

Article Title: Differential regulation of radioadaptation by quercetin between human normal and cancer cells

doi: 10.1016/j.ctro.2025.101099

Figure Lengend Snippet: Quercetin restores NRF2 nuclear translocation in radioadapted MCF10A cells. Representative immunofluorescence images of NRF2 (green) and DAPI (blue) and quantification of the nuclear-to-cytoplasmic NRF2 fluorescence ratio in MCF10A cells 24 h after 5 Gy irradiation with or without prior LDRT and quercetin treatment (Scale bar: 10 μm). Data is shown as mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were incubated with rabbit anti-human NRF2 primary antibody (1:200; Proteintech, 16396–1-AP), and detection was performed using goat anti-rabbit Alexa Fluor 488 (1:500, Invitrogen, 11001).

Techniques: Translocation Assay, Immunofluorescence, Fluorescence, Irradiation

Journal: Aging Cell

Article Title: Overactivation of Cdc42 GTPase Impairs the Cytotoxic Function of NK Cells From Old Individuals Towards Senescent Fibroblasts

doi: 10.1111/acel.70398

Figure Lengend Snippet: Natural killer cells from old adults reveal reduced cytotoxicity towards senescent fibroblasts. (A) Graphical illustration of the NK cell mediated target cell cytotoxicity assay. Target cells were first stained with calcein acetomethoxymethyl (AM), a vital fluorescent dye. Calcein AM is a non‐fluorescent compound that pass the intact cell membrane into the cytoplasm. Hydrolysis of calcein AM by intracellular esterases in live cells generates calcein, a hydrophilic, intensely fluorescent molecule which reliably stays in the cytoplasm. The stained target cells were next co‐cultured with NK cells isolated from young or old human or mice. NK cells exert their cytotoxicity towards target cells through the release of perforin and granzyme B. Upon lysis of target cells, the calcein dye is released and the loss of the dye is measured as a shift in fluorescence intensity by flow cytometry. Dead cells will appear to the left of the histogram, while alive cells on the right side. The percentage of dead cells can then simply be calculated and presented. (B) Graphical scheme depicts the experimental groups: Co‐cultures of NK cells from young adults with senescent human dermal fibroblasts in the top row and NK cells from old adults with senescent HDF in the bottom row. (C) Histogram (bi‐exponential scale) showing cytotoxicity of NK cells from young and old adults on different senescent HDF. RS, replicative senescent HDF, DIS, doxorubicin induced senescent HDF, IR, ionizing radiation induced senescence, CA, chronologically aged HDF (~75 years). The peak in the left part of the histogram showing the dead cell population and the percentage of dead cells. (D) The graph depicts the percentage of senescent HDF death ( y ‐axis) by NK cells isolated from young and old human adult. Data were represented as mean (Percentage of senescent fibroblast death) ± SEM. N = 5. Two tailed Student's t ‐test was used to assess the significance between young and old groups for each of senescence model. (E) Illustration of the experimental design showing cytotoxic activity of NK cells derived from bone marrow and spleen of young and old mice against aged murine dermal fibroblasts (MDF). (F) Histogram depicting cytotoxicity of NK cells from young and old mice on old MDF. The peak in the left part of the histogram showing the dead cell population and the percentage of dead cells. (G) The graph depicts the percentage of senescent MDF death ( y ‐axis) by NK cells isolated from young and old mice. Data were represented as mean (Percentage of senescent fibroblast death) ± SEM. N = 3. Each mouse NK cell sample used in the cytotoxicity assay was the pool of NK cells isolated from three different mice. One‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups.

Article Snippet: Human dermal fibroblast , ATCC , Cat. #ATCC‐PCS‐201‐010.

Techniques: Cytotoxicity Assay, Staining, Membrane, Cell Culture, Isolation, Lysis, Fluorescence, Flow Cytometry, Two Tailed Test, Activity Assay, Derivative Assay, Comparison

Journal: Aging Cell

Article Title: Overactivation of Cdc42 GTPase Impairs the Cytotoxic Function of NK Cells From Old Individuals Towards Senescent Fibroblasts

doi: 10.1111/acel.70398

Figure Lengend Snippet: CASIN restores impairment of conjugation, degranulation and mitochondrial ATP generation in old Natural killer cells. (A) Synapse formation with conjugation of NK cells with the target senescent fibroblasts and the tubulin network pulling the perforin and granzyme B containing vesicles in the direction of the synapse. (B) Fusion of the NK cell derived secretory granules with the presynaptic membrane of NK cells and concomitant exposure of CD107a at the cell membrane and the release of perforin and granzyme B into the synaptic cleft towards the target cell. (C & D) Percentage of NK cell conjugation with senescent HDF when co‐cultured for (C) 60 and (D) 90 min at an effector to target (E:T) cell ratio of 1:1. Data were represented as mean (percentage of cell conjugation) ± SEM, N = 3. (E) Degranulation of NK cells when co‐cultured with senescent HDF for 7 h at an effector to target (E:T) cell ratio of 10:1. Data were represented as mean (mean fluorescence intensity) ± SEM, N = 6. One‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups in C, D and E. (F) Seahorse flux analysis showed quantification of ATP generated by NK cells treated with vehicle from young donors and from NK cells treated with either vehicle or CASIN from old donors. The ATP generation either by glycolysis or by oxidative phosphorylation and total ATP was assessed. Data were represented as mean (ATP level) ± SEM, N = 7. Two‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups. (G) Mitochondrial structure showing the chemical structure of the mitochondrial fluorescent probe JC‐1 that can form J‐aggregates (red) and J‐monomers (green) indicating high and low mitochondrial membrane potential, respectively. (H) Flow cytometry analysis of J‐aggregates (red) and J‐monomers (green) of young NK cells treated with vehicle, old NK cells treated with either vehicle or CASIN. (I) The graph depicts the percentage of J‐aggregates (Q2 population of figure H) of young NK treated with vehicle, and old NK cells treated with either vehicle or CASIN. Data were represented as mean (percentage of cell with J‐aggregate) ± SEM, N = 6. (J) Quantification of the ratio of J‐aggregates to J‐monomers from young and old NK treated with vehicle and old NK cells treated with CASIN. Data were represented as mean (ratio) ± SEM, N = 5. One‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups in I and J.

Article Snippet: Human dermal fibroblast , ATCC , Cat. #ATCC‐PCS‐201‐010.

Techniques: Conjugation Assay, Derivative Assay, Membrane, Cell Culture, Fluorescence, Comparison, Generated, Phospho-proteomics, Flow Cytometry

Journal: Aging Cell

Article Title: Overactivation of Cdc42 GTPase Impairs the Cytotoxic Function of NK Cells From Old Individuals Towards Senescent Fibroblasts

doi: 10.1111/acel.70398

Figure Lengend Snippet: CASIN treatment improves the cytotoxic ability of Natural killer cells from old humans and mice. (A) Graphical illustration of experimental plan, where young NK cells treated with vehicle and old NK cells treated with either vehicle or CASIN for 8 h and thereafter subjected to co‐culture with target senescent HDF exerting their differential killing ability. (B) Representative histograms depicting the killing ability of different experimental groups as measured by flow cytometry. Peak at the left side of histogram, showing the dead senescent HDF population with percentage of dead cells. (C) Quantification of the percentage of target senescent HDF death executed by young NK cells treated with vehicle, and old NK cells treated with either vehicle or CASIN. Data were represented as mean (Percentage of senescent fibroblast death) ± SEM. N = 4. (D) Representative histograms show the distribution of K562 killing by young NK cells treated with vehicle, and old NK cells treated with either vehicle or CASIN. Peak at the left side of histogram, showing the dead K562 population with percentage of dead cells. (E) Graph shows the percentage of target cell (K562) death mediated either by young NK cells treated with vehicle or by old NK cells treated with either vehicle or CASIN. Data were represented as mean (Percentage of K562 lysis) ± SEM. N = 4. One‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups in C and E. (F) Illustration of the experimental design for treatment of young mice (average age 120 days) treated with vehicle and old mice (average age 650 days) treated with either vehicle or CASIN. Following treatment, NK cells were isolated from spleen and bone marrow and subjected to co‐cultures with murine dermal fibroblasts (MDF) derived from old mice (average age 650 days). (G) Flow cytometry with representative histograms depicting old/senescent MDF killing by NK cells isolated from bone marrow (left panel) and spleen (right panel) of vehicle and CASIN treated old mice. Peak at the left side of histogram, showing the dead old MDF population with percentage of dead cells. (H) Quantification of the percentage of old/senescent MDF killing by NK cells isolated from bone marrow and spleen of young and old mice treated with vehicle and old mice treated with CASIN. Data were represented as mean (percentage of old/senescent MDF lysis) ± SEM, N = 4, where each group contains pool of NK cells isolated from 4 different mice of same treatment group. Two‐way ANOVA, followed by Bonferroni multiple comparison test was used to find the significance among the groups. (I) Graphical summary. Unrestrained Cdc42 activity causes failure of old NK cells to kill senescent fibroblasts. Unrestrained Cdc42 activity disrupts the microtubular network and impaired mitochondrial ATP resulting in reduced conjugation, and impaired degranulation of lytic vesicles into the synaptic cleft with reduced cytotoxicity. CASIN can attenuate all these steps and in part attenuate the killing of senescent fibroblasts (senescent HDF).

Article Snippet: Human dermal fibroblast , ATCC , Cat. #ATCC‐PCS‐201‐010.

Techniques: Co-Culture Assay, Flow Cytometry, Lysis, Comparison, Isolation, Derivative Assay, Activity Assay, Conjugation Assay

Journal: Scientific Reports

Article Title: Chitosan-dextran sulfate nanocapsules for enhanced tigecycline efficacy against non-typhoidal Salmonella enterica

doi: 10.1038/s41598-026-35229-7

Figure Lengend Snippet: TGC and/or unloaded CD NPs ameliorated the S. Typhimurium infected group induced histopathological alterations in mice’s liver tissues. Representative photomicrographs of the H&E-stained hepatic tissue sections showing the control (A), S. Typhimurium infected group (C), TGC (E), unloaded CD NPs (G), CD-TGC groups (I) and their respective higher magnifications (B, D, F, H, and J). A, B: control group displaying normal central vein (CV), hepatic cords (HC) with hepatocytes of eosinophilic granular cytoplasm (EC), rounded central single (SN) or double vesicular nuclei (DN), and kupffer cells (KC). C, D: S. Typhimurium infected group demonstrating multiple areas of variable-sized necrotic areas (NA) of coagulative necrosis (CN), severely dilated and congested sinusoid (SDS) with Kupffer cell hyperplasia (KCH). E, F: TGC group displaying moderately sized necrotic areas (MNA), moderately dilated and congested sinusoids (MDS), moderately hyperplastic Kupffer’s cells (MKC), interstitial mononuclear cell infiltration (MI), fibroblast proliferation (FP), and regenerated hepatocytes of stippling basophilic cytoplasm (BC) and pale nuclei (PN). G, H: unloaded CD NPs group showing a few scattered minute necrotic areas (mNA), intense mononuclear cell infiltration (IMI) around the portal area, and apparently normal hepatocytes (NH). I, J: CD-TGC group showed normal hepatocytes (NH), few dilated blood vessels (DBV), and a few interstitial lymphocytic aggregates (FL). Scale bars = 100 μm in A, C, E, G, I; and = 20 μm in B, D, F, H, and J. K: Bar charts demonstrate the statistical analysis of the comparative quantification of the hepatic injury scores in all studied groups. Bars carrying different superscript letters (a, b, c, d, and e) are significantly different as analyzed by the one-way ANOVA test, followed by the multiple comparisons by Duncan’s Post-hoc test ( p < 0.05). Values are the mean of 6 mice per group ± S.E.M.

Article Snippet: BJ normal human foreskin primary fibroblast cell line (ATCC CRL-2522) was used for studying safety of CD-TGC nanocapsules.

Techniques: Infection, Staining, Control