nmol l tumor necrosis factor  (R&D Systems)

Journal: Bioactive Materials

Article Title: Inhalable PD-L1-engineered hybrid cellular vesicles suppress excessive neutrophil activation and restore mitochondrial homeostasis to alleviate ischemia–reperfusion lung injury and pneumonia

doi: 10.1016/j.bioactmat.2026.03.024

Figure Lengend Snippet: Characterization of Res-PD-L1@nmEVs . (A) Schematic illustration of the Res-PD-L1@nmEVs synthesis procedure. (B-D) Representative transmission electron microscopy (TEM) images, dynamic light scattering (DLS) size distributions, and zeta potential measurements of nEVs, PD-L1@mEVs, PD-L1@nmEVs, and Res-PD-L1@nmEVs. (E) PD-L1 expression in PD-L1-overexpressing MSCs (OE-PD-L1) and negative control (NC) MSCs, and CD11b expression in HL60 cells before and after DMSO stimulation, as determined by Western blot. (F) Expression levels of neutrophil membrane markers (CD11b, CXCR2, RAGE, TLR2) and the exosomal marker CD63 in the four EV types. (G) Fluorescence co-localization images of DiO-labeled nEVs (green) and DiL-labeled PD-L1@mEVs (red) after fusion, demonstrating hybrid vesicle formation. (H) Size stability of Res-PD-L1@nmEVs stored at 4 °C and 37 °C for 7 days. (I-K) Binding and neutralization capacity of Res-PD-L1@nmEVs against inflammatory cytokines (TNF-α, IL-6, IL-1β) in vitro. ∗ vs. 0ug/ml; # vs. 100 μg/ml, p < 0.05, n = 5.

Article Snippet: These cells were then stimulated with 40 ng/mL recombinant TNF-α (C008, Novoprotein, China) for 6 h to induce an N1-type neutrophil phenotype.

Techniques: Transmission Assay, Electron Microscopy, Zeta Potential Analyzer, Expressing, Negative Control, Western Blot, Membrane, Marker, Fluorescence, Labeling, Binding Assay, Neutralization, In Vitro

Journal: Bioactive Materials

Article Title: Inhalable PD-L1-engineered hybrid cellular vesicles suppress excessive neutrophil activation and restore mitochondrial homeostasis to alleviate ischemia–reperfusion lung injury and pneumonia

doi: 10.1016/j.bioactmat.2026.03.024

Figure Lengend Snippet: Res-PD-L1@nmEVs Attenuate Inflammation and Oxidative Damage in Lung Epithelial Cells In Vitro . (A-B) Flow cytometric analysis and quantification (B) of DiO-labeled Res-PD-L1@nmEVs uptake by BEAS-2B cells under H/R conditions after pretreatment with different endocytic inhibitors (chlorpromazine, chloroquine, and filipin) or incubation at 4 °C. (C) mRNA expression levels of IL-6, TNF-α, and IL-1β in BEAS-2B cells with or without H/R injury following pretreatment with Res, nEVs, PD-L1@mEVs, PD-L1@nmEVs, or Res-PD-L1@nmEVs. (D-E) Representative fluorescence images (D) and quantitative analysis (E) of cell proliferation assessed by BrdU incorporation (red; nuclei stained with DAPI, blue). Scale bar: 50 μm. (F-G) Apoptosis rates detected by flow cytometry (F) and flow cytometric analysis of Annexin V-positive BEAS-2B cells under the indicated conditions (G). (H–K) Fluorescence microscopy images and quantitative analysis of intracellular nitric oxide (NO, green) (H-I) and reactive oxygen species (ROS, red) (J-K). Scale bar: 100 μm. (L) Flow cytometry analysis of intracellular ROS levels. (M − O) Levels of malondialdehyde (MDA) (M), superoxide dismutase 2 (SOD2) activity (N), and glutathione (GSH) content (O) in cells. (P-Q) Cell migration ability evaluated by wound healing assay under different treatments. ∗ vs. Control; # vs. H/R; & vs. H/R + PD-L1@nmEVs, p < 0.05.

Article Snippet: These cells were then stimulated with 40 ng/mL recombinant TNF-α (C008, Novoprotein, China) for 6 h to induce an N1-type neutrophil phenotype.

Techniques: In Vitro, Labeling, Incubation, Expressing, Fluorescence, BrdU Incorporation Assay, Staining, Flow Cytometry, Microscopy, Activity Assay, Migration, Wound Healing Assay, Control

Journal: Bioactive Materials

Article Title: Inhalable PD-L1-engineered hybrid cellular vesicles suppress excessive neutrophil activation and restore mitochondrial homeostasis to alleviate ischemia–reperfusion lung injury and pneumonia

doi: 10.1016/j.bioactmat.2026.03.024

Figure Lengend Snippet: Res-PD-L1@nmEVs Suppresses Neutrophil Activation HL60 cells were differentiated into neutrophil-like cells using DMSO and subsequently stimulated with TNF-α to induce activation under conditions simulating IRI. The effects of Res, nEVs, PD-L1@mEVs, PD-L1@nmEVs, and Res-PD-L1@nmEVs on neutrophil activation were evaluated. (A) Cell surface PD-1 expression analyzed by flow cytometry. (B) Representative immunofluorescence images of CD206 expression (red). Nuclei were stained with DAPI (blue). Scale bar: 50 μm. (C) Flow cytometric analysis of cell surface CD206 expression. (D) Flow cytometric analysis of cell surface CD95 expression. (E-G) Levels of myeloperoxidase (MPO) (E), neutrophil elastase (NE) (F), and MMP-9 (G) in neutrophil culture supernatants, measured by ELISA. (H-J) BEAS-2B cells were co-cultured with neutrophils in the presence or absence of TNF-α stimulation. Apoptosis levels (I) and migration capacity (J) of BEAS-2B cells were assessed under different treatment conditions. ∗ vs. Control; # vs. TNF-a; & vs. TNF-a+PD-L1@nmEVs, p < 0.05.

Article Snippet: These cells were then stimulated with 40 ng/mL recombinant TNF-α (C008, Novoprotein, China) for 6 h to induce an N1-type neutrophil phenotype.

Techniques: Activation Assay, Expressing, Flow Cytometry, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture, Migration, Control

Journal: Bioactive Materials

Article Title: Inhalable PD-L1-engineered hybrid cellular vesicles suppress excessive neutrophil activation and restore mitochondrial homeostasis to alleviate ischemia–reperfusion lung injury and pneumonia

doi: 10.1016/j.bioactmat.2026.03.024

Figure Lengend Snippet: Res-PD-L1@nmEVs Effectively Attenuates MRSA-Induced Pneumonia (A-B) Rats with MRSA-induced pneumonia received three bronchial nebulization treatments over one week with different formulations (Res, nEVs, PD-L1@mEVs, PD-L1@nmEVs, or Res-PD-L1@nmEVs). (A) Representative H&E-stained lung sections and (B) corresponding lung injury scores are shown (n = 5). (C) TUNEL staining of lung tissues to assess apoptosis. (D) Representative micro-CT images of anesthetized rats. (E-G) Flow cytometric analysis of immune cell proportions in lung single-cell suspensions: CD8 + T cells (E), neutrophils (F), and classical monocytes (G). (H-J) Plasma levels of inflammatory cytokines IL-6 (H), IL-1β (I), and TNF-α (J) (n = 5). (K) Immunofluorescence staining of tight junction proteins Occludin (green) and ZO-1 (red) in lung tissues (nuclei stained with DAPI). Scale bar: 50 μm. (L-N) Pulmonary function parameters: lung compliance (L), airway resistance (M), and oxygenation index (N) (n = 4). ∗ vs. Sham; # vs. MRSA; & vs. MRSA + PD-L1@nmEVs, p < 0.05.

Article Snippet: These cells were then stimulated with 40 ng/mL recombinant TNF-α (C008, Novoprotein, China) for 6 h to induce an N1-type neutrophil phenotype.

Techniques: Staining, TUNEL Assay, Micro-CT, Single Cell, Clinical Proteomics, Immunofluorescence

Journal: Poultry Science

Article Title: Pathogen identification and analysis of broiler bronchial obstruction syndrome using high-throughput sequencing technology

doi: 10.1016/j.psj.2026.107032

Figure Lengend Snippet: Recombination analysis of CK/CH/HBBD/BD6 and CK/CH/HBBD/BD3. The genomic characteristics of IBV and the whole-genome recombination analysis were conducted using the Simplot software for Bootscan analysis (window size: 200 bp, step size: 20 bp).

Article Snippet: Bootscan analysis performed using Simplot software ( ) further validated the recombination events identified by RDP4 software and no recombination events were found in the S gene.

Techniques: Software

Journal: Poultry Science

Article Title: Immunocastration with recombinant GnRH6-CRM197 fusion protein improves reproductive suppression and meat tenderness in roosters

doi: 10.1016/j.psj.2026.107046

Figure Lengend Snippet: SDS-PAGE analysis of expression of pET21a-GnRH6-CRM197 in E. coli. BL21. Lane 1, standard molecular weight marker; Lane 2, non-induced recombinant bacteria; Lane 3, recombinant bacteria induced after 12 h.

Article Snippet: The GnRH6-CRM197 recombinant protein was separated on a 12.5% SDS polyacrylamide gel at 120 V. It was then transferred to a 0.45μm polyvinylidene fluoride (PVDF) membrane (Pulilai, Beijing, China) at 250 mA for 1 hour using a transfer system (BVT-2, Servicebio, Wuhan, China).

Techniques: SDS Page, Expressing, Molecular Weight, Marker, Recombinant, Bacteria

Journal: Poultry Science

Article Title: Immunocastration with recombinant GnRH6-CRM197 fusion protein improves reproductive suppression and meat tenderness in roosters

doi: 10.1016/j.psj.2026.107046

Figure Lengend Snippet: WB analysis of recombinant GnRH6-CRM197 protein. Lane 1, standard molecular weight marker; Lane 2, recombinant GnRH6-CRM197 fusion protein reacted with rabbit polyclonal to GnRH (dilution 1:2000) .

Article Snippet: The GnRH6-CRM197 recombinant protein was separated on a 12.5% SDS polyacrylamide gel at 120 V. It was then transferred to a 0.45μm polyvinylidene fluoride (PVDF) membrane (Pulilai, Beijing, China) at 250 mA for 1 hour using a transfer system (BVT-2, Servicebio, Wuhan, China).

Techniques: Recombinant, Molecular Weight, Marker

Journal: Poultry Science

Article Title: Immunocastration with recombinant GnRH6-CRM197 fusion protein improves reproductive suppression and meat tenderness in roosters

doi: 10.1016/j.psj.2026.107046

Figure Lengend Snippet: The effect of recombinant anti-GnRH6-CRM197 on serum anti-GnRH antibody of roosters. The arrow refers to the immune time; the first immunity of roosters is 4 W; * P < 0.05; ** P < 0.01 (n = 20 per group).

Article Snippet: The GnRH6-CRM197 recombinant protein was separated on a 12.5% SDS polyacrylamide gel at 120 V. It was then transferred to a 0.45μm polyvinylidene fluoride (PVDF) membrane (Pulilai, Beijing, China) at 250 mA for 1 hour using a transfer system (BVT-2, Servicebio, Wuhan, China).

Techniques: Recombinant

Journal: Poultry Science

Article Title: Immunocastration with recombinant GnRH6-CRM197 fusion protein improves reproductive suppression and meat tenderness in roosters

doi: 10.1016/j.psj.2026.107046

Figure Lengend Snippet: The effect of recombinant GnRH6-CRM197 on serum testosterone levels of roosters. * P < 0.05; ** P < 0.01 (n = 20 per group).

Article Snippet: The GnRH6-CRM197 recombinant protein was separated on a 12.5% SDS polyacrylamide gel at 120 V. It was then transferred to a 0.45μm polyvinylidene fluoride (PVDF) membrane (Pulilai, Beijing, China) at 250 mA for 1 hour using a transfer system (BVT-2, Servicebio, Wuhan, China).

Techniques: Recombinant