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Concentration-dependent change in the humoral inflammatory response following incubation with Escherichia coli ( E. coli ) in the ex vivo whole blood model. a Absolute plasma concentrations of IL-6, <t>IL-8,</t> and MMP9 determined by enzyme-linked immunosorbent assay. b Normalized values and EC 50 curve fit by BuC=0% and 50 000 CFU/ml E. coli= 100%, respectively, for IL-6, IL-8, and MMP9 as indicated by EC 50 (%) on the respective Y-axis. BuC indicates buffer control after 60 min incubation; numbers on the X-axis indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation; LPS indicates lipopolysaccharide (LPS) 100 ng/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, comparing all shown concentrations of E. coli bacteria and 100 ng/ml LPS with BuC. P -values are indicated above the respective data points. ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.
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circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of <t>IL-10,</t> TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.
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circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of <t>IL-10,</t> TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.
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Angiogenesis and collagen deposition in diabetic wound tissues following HPSL@SG hydrogel treatment. (A) Dihydroethidium (DHE) immunofluorescence staining and (B) semi-quantitative analysis of wound tissues from each treatment group on day 7, scale bar = 100 μm. Immunofluorescence staining of (C) MMP-9, IL-6, and <t>IL-10,</t> and (D) CD31, VEGF-A, and collagen I in wound tissue sections from each treatment group on day 7, scale bar = 100 μm. (E-J) Mean relative fluorescence intensity of each indicator in wound tissue sections from each treatment group on day 7, scale bar = 100 μm. All data are shown as mean ± SEM (n = 6).
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circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of <t>IL-10,</t> TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.
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Elevated PD-1 expression and altered cytokine levels in plasma of IE patients. (A) PD-1 expression on CD4 + CD25 high Tregs is significantly increased in IE patients, particularly in the ISE subgroup, while CD4 + and CD4 + CD25 + T cell percentages are reduced. (B) Plasma concentrations <t>of</t> <t>IL-10</t> and IL-6 are elevated in IE patients, with IL-10 showing a progressive increase with seizure severity. (C) Correlation analyses reveal associations between immune parameters and clinical characteristics. ∗∗∗P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05; ns indicates not significant.
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GMI gel -mediated metabolic reprogramming and its regulatory role in macrophage polarization. A-D) Hexokinase activity, Phosphofructokinase activity, Isocitrate dehydrogenase activity, and Succinate dehydrogenase activity, n = 3. E-H) Heatmap of LC-MS data and quantitative analysis of the relative abundance of glycolysis and TCA cycle metabolites, n = 3. I) Schematic diagram of glycolysis and the TCA cycle. J) Representative CLSM images of RAW 264.7 cells treated with GMI gel for 72 h. K) Flow cytometry analysis of CD206 and CD86 expression. L, M) The secretion <t>of</t> <t>IL-6</t> and TNF-α in the culture supernatant of RAW264.7 cells, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns, not significant (p > 0.05).
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GMI gel -mediated metabolic reprogramming and its regulatory role in macrophage polarization. A-D) Hexokinase activity, Phosphofructokinase activity, Isocitrate dehydrogenase activity, and Succinate dehydrogenase activity, n = 3. E-H) Heatmap of LC-MS data and quantitative analysis of the relative abundance of glycolysis and TCA cycle metabolites, n = 3. I) Schematic diagram of glycolysis and the TCA cycle. J) Representative CLSM images of RAW 264.7 cells treated with GMI gel for 72 h. K) Flow cytometry analysis of CD206 and CD86 expression. L, M) The secretion <t>of</t> <t>IL-6</t> and TNF-α in the culture supernatant of RAW264.7 cells, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns, not significant (p > 0.05).
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Concentration-dependent change in the humoral inflammatory response following incubation with Escherichia coli ( E. coli ) in the ex vivo whole blood model. a Absolute plasma concentrations of IL-6, IL-8, and MMP9 determined by enzyme-linked immunosorbent assay. b Normalized values and EC 50 curve fit by BuC=0% and 50 000 CFU/ml E. coli= 100%, respectively, for IL-6, IL-8, and MMP9 as indicated by EC 50 (%) on the respective Y-axis. BuC indicates buffer control after 60 min incubation; numbers on the X-axis indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation; LPS indicates lipopolysaccharide (LPS) 100 ng/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, comparing all shown concentrations of E. coli bacteria and 100 ng/ml LPS with BuC. P -values are indicated above the respective data points. ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Concentration-dependent change in the humoral inflammatory response following incubation with Escherichia coli ( E. coli ) in the ex vivo whole blood model. a Absolute plasma concentrations of IL-6, IL-8, and MMP9 determined by enzyme-linked immunosorbent assay. b Normalized values and EC 50 curve fit by BuC=0% and 50 000 CFU/ml E. coli= 100%, respectively, for IL-6, IL-8, and MMP9 as indicated by EC 50 (%) on the respective Y-axis. BuC indicates buffer control after 60 min incubation; numbers on the X-axis indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation; LPS indicates lipopolysaccharide (LPS) 100 ng/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, comparing all shown concentrations of E. coli bacteria and 100 ng/ml LPS with BuC. P -values are indicated above the respective data points. ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Concentration Assay, Incubation, Ex Vivo, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Bacteria

Diagnostic performance for the detection of bacteremia, analyzing the neutrophil phenotype by determining the median fluorescence intensity (MFI) and the cellular response capacity (CRC) in comparison with traditional markers of humoral inflammation (IL-6, IL-8, MMP9). a Comparison of receiver operating characteristic (ROC) at 10,000 CFU/ml Escherichia coli ( E. coli ) with the respective 95% confidence interval (CI) and P -value, and half-maximal effective concentration (EC 50 ) as a function of the E. coli concentration. b Detailed comparison of the EC 50 as a function of the E. coli concentration. c Exemplary comparison of EC 50 curve fit after normalization as indicated by EC 50 (%) on the respective Y-axis to BuC=100% and 50 000 CFU/ml E. coli =0% for the humoral marker IL-6 (the IL-6 values were multiplied by −1 before EC 50 calculation to facilitate comparability with the CRC) and the change in neutrophil phenotype represented by CD11b CRC. BuC indicates buffer control after 60 min incubation; numbers on the X-axis of c indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, evaluating the EC 50 of IL-8, MMP9, the MFI, and CRC of CD10, CD11b, and CD62L in comparison to the EC 50 of IL-6. P -values are indicated above the respective data points. ⁎ P <0.05. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Diagnostic performance for the detection of bacteremia, analyzing the neutrophil phenotype by determining the median fluorescence intensity (MFI) and the cellular response capacity (CRC) in comparison with traditional markers of humoral inflammation (IL-6, IL-8, MMP9). a Comparison of receiver operating characteristic (ROC) at 10,000 CFU/ml Escherichia coli ( E. coli ) with the respective 95% confidence interval (CI) and P -value, and half-maximal effective concentration (EC 50 ) as a function of the E. coli concentration. b Detailed comparison of the EC 50 as a function of the E. coli concentration. c Exemplary comparison of EC 50 curve fit after normalization as indicated by EC 50 (%) on the respective Y-axis to BuC=100% and 50 000 CFU/ml E. coli =0% for the humoral marker IL-6 (the IL-6 values were multiplied by −1 before EC 50 calculation to facilitate comparability with the CRC) and the change in neutrophil phenotype represented by CD11b CRC. BuC indicates buffer control after 60 min incubation; numbers on the X-axis of c indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, evaluating the EC 50 of IL-8, MMP9, the MFI, and CRC of CD10, CD11b, and CD62L in comparison to the EC 50 of IL-6. P -values are indicated above the respective data points. ⁎ P <0.05. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Diagnostic Assay, Fluorescence, Comparison, Concentration Assay, Marker, Control, Incubation, Bacteria

Clinical specifications and parameters over all time points of the sepsis cohort. a Suspected infection cause of sepsis. b Distribution of the individual score points of the Sequential Organ Failure Assessment (SOFA) score. c Total SOFA score. d-h Traditional and humoral markers of inflammation: leukocytes and neutrophil-lymphocyte ratio ( d ), C-reactive protein (CRP) and procalcitonin (PCT) ( e ), interleukin-6 (IL-6) and interleukin-8 (IL-8) ( f ), serum amyloid A (SAA) and calprotectin ( g ), matrix metallopeptidase 9 (MMP9) and myeloperoxidase (MPO) ( h ). Values are shown as median and interquartile range. n =14. CNS. Central nervous system; HV. Healthy volunteers.

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Clinical specifications and parameters over all time points of the sepsis cohort. a Suspected infection cause of sepsis. b Distribution of the individual score points of the Sequential Organ Failure Assessment (SOFA) score. c Total SOFA score. d-h Traditional and humoral markers of inflammation: leukocytes and neutrophil-lymphocyte ratio ( d ), C-reactive protein (CRP) and procalcitonin (PCT) ( e ), interleukin-6 (IL-6) and interleukin-8 (IL-8) ( f ), serum amyloid A (SAA) and calprotectin ( g ), matrix metallopeptidase 9 (MMP9) and myeloperoxidase (MPO) ( h ). Values are shown as median and interquartile range. n =14. CNS. Central nervous system; HV. Healthy volunteers.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Infection

circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: For mouse experiments, mouse IL-10 was measured using the Mouse IL-10 ELISA Kit (R&D Systems, Cat# M1000B), and mouse TGF-β1 was measured using the Mouse TGF beta-1 ELISA Kit (Invitrogen, Cat# BMS608-4), following the manufacturers’ instructions.

Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration

circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Human IL-10 was quantified using IL-10 DuoSet ELISA (R&D Systems, Cat# DY217B), and human TGF-β1 was measured using the Human TGF beta-1 ELISA Kit (Invitrogen, Cat# BMS249-4).

Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration

Angiogenesis and collagen deposition in diabetic wound tissues following HPSL@SG hydrogel treatment. (A) Dihydroethidium (DHE) immunofluorescence staining and (B) semi-quantitative analysis of wound tissues from each treatment group on day 7, scale bar = 100 μm. Immunofluorescence staining of (C) MMP-9, IL-6, and IL-10, and (D) CD31, VEGF-A, and collagen I in wound tissue sections from each treatment group on day 7, scale bar = 100 μm. (E-J) Mean relative fluorescence intensity of each indicator in wound tissue sections from each treatment group on day 7, scale bar = 100 μm. All data are shown as mean ± SEM (n = 6).

Journal: Bioactive Materials

Article Title: Glucose/ROS-responsive and redox-gated adaptive hydrogel dressing for accelerating diabetic wound repair via synergistic cGAS/STING pathway inhibition and oxidative stress alleviation

doi: 10.1016/j.bioactmat.2026.03.025

Figure Lengend Snippet: Angiogenesis and collagen deposition in diabetic wound tissues following HPSL@SG hydrogel treatment. (A) Dihydroethidium (DHE) immunofluorescence staining and (B) semi-quantitative analysis of wound tissues from each treatment group on day 7, scale bar = 100 μm. Immunofluorescence staining of (C) MMP-9, IL-6, and IL-10, and (D) CD31, VEGF-A, and collagen I in wound tissue sections from each treatment group on day 7, scale bar = 100 μm. (E-J) Mean relative fluorescence intensity of each indicator in wound tissue sections from each treatment group on day 7, scale bar = 100 μm. All data are shown as mean ± SEM (n = 6).

Article Snippet: IL-6 and IL-10-specific antibodies were purchased from Bosterbio (Wuhan, China).

Techniques: Immunofluorescence, Staining, Fluorescence

circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Human IL-10 was quantified using IL-10 DuoSet ELISA (R&D Systems, Cat# DY217B), and human TGF-β1 was measured using the Human TGF beta-1 ELISA Kit (Invitrogen, Cat# BMS249-4).

Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration

Elevated PD-1 expression and altered cytokine levels in plasma of IE patients. (A) PD-1 expression on CD4 + CD25 high Tregs is significantly increased in IE patients, particularly in the ISE subgroup, while CD4 + and CD4 + CD25 + T cell percentages are reduced. (B) Plasma concentrations of IL-10 and IL-6 are elevated in IE patients, with IL-10 showing a progressive increase with seizure severity. (C) Correlation analyses reveal associations between immune parameters and clinical characteristics. ∗∗∗P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05; ns indicates not significant.

Journal: Brain, Behavior, & Immunity - Health

Article Title: Exploring the role of PD-1 as a marker in drug-refractory epilepsy and its potential indication for valproic acid treatment

doi: 10.1016/j.bbih.2026.101238

Figure Lengend Snippet: Elevated PD-1 expression and altered cytokine levels in plasma of IE patients. (A) PD-1 expression on CD4 + CD25 high Tregs is significantly increased in IE patients, particularly in the ISE subgroup, while CD4 + and CD4 + CD25 + T cell percentages are reduced. (B) Plasma concentrations of IL-10 and IL-6 are elevated in IE patients, with IL-10 showing a progressive increase with seizure severity. (C) Correlation analyses reveal associations between immune parameters and clinical characteristics. ∗∗∗P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05; ns indicates not significant.

Article Snippet: Concentrations of IL-10 and IL-6 in plasma and CSF were quantified using commercial double-antibody sandwich ELISA kits (Boster Bioengineering, China), according to the manufacturer's instructions.

Techniques: Expressing, Clinical Proteomics

Central nervous system immune activation in ISE patients. (A) PD-1 + CD4 + CD25 high Treg cell counts are significantly elevated in the CSF of ISE patients compared to controls. (B) Both IL-10 and IL-6 concentrations are increased in the CSF of ISE patients. (C) Age correlates positively with PD-1 + CD4 + CD25 low and CD4 + CD25 high Treg percentages, and negatively with CD4 + T cell percentage in the CSF of ISE patients. ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05.

Journal: Brain, Behavior, & Immunity - Health

Article Title: Exploring the role of PD-1 as a marker in drug-refractory epilepsy and its potential indication for valproic acid treatment

doi: 10.1016/j.bbih.2026.101238

Figure Lengend Snippet: Central nervous system immune activation in ISE patients. (A) PD-1 + CD4 + CD25 high Treg cell counts are significantly elevated in the CSF of ISE patients compared to controls. (B) Both IL-10 and IL-6 concentrations are increased in the CSF of ISE patients. (C) Age correlates positively with PD-1 + CD4 + CD25 low and CD4 + CD25 high Treg percentages, and negatively with CD4 + T cell percentage in the CSF of ISE patients. ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05.

Article Snippet: Concentrations of IL-10 and IL-6 in plasma and CSF were quantified using commercial double-antibody sandwich ELISA kits (Boster Bioengineering, China), according to the manufacturer's instructions.

Techniques: Activation Assay

VPA treatment modulates PD-1 expression and IL-10 levels independently of drug concentration. (A) Plasma PD-1 expression on both CD4 + CD25 high and CD4 + CD25 low Treg subsets decreases significantly after 48 h of VPA treatment. (B) Plasma IL-10 levels decrease significantly after VPA treatment, while IL-6 shows no significant change. (C) Seizure control time correlates with immunological parameters but not with VPA concentrations. (D) VPA concentrations in CSF are significantly lower than in plasma, with no change between d0 and d3, and no correlation with seizure control time. ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05; ns indicates not significant.

Journal: Brain, Behavior, & Immunity - Health

Article Title: Exploring the role of PD-1 as a marker in drug-refractory epilepsy and its potential indication for valproic acid treatment

doi: 10.1016/j.bbih.2026.101238

Figure Lengend Snippet: VPA treatment modulates PD-1 expression and IL-10 levels independently of drug concentration. (A) Plasma PD-1 expression on both CD4 + CD25 high and CD4 + CD25 low Treg subsets decreases significantly after 48 h of VPA treatment. (B) Plasma IL-10 levels decrease significantly after VPA treatment, while IL-6 shows no significant change. (C) Seizure control time correlates with immunological parameters but not with VPA concentrations. (D) VPA concentrations in CSF are significantly lower than in plasma, with no change between d0 and d3, and no correlation with seizure control time. ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05; ns indicates not significant.

Article Snippet: Concentrations of IL-10 and IL-6 in plasma and CSF were quantified using commercial double-antibody sandwich ELISA kits (Boster Bioengineering, China), according to the manufacturer's instructions.

Techniques: Expressing, Concentration Assay, Clinical Proteomics, Control

GMI gel -mediated metabolic reprogramming and its regulatory role in macrophage polarization. A-D) Hexokinase activity, Phosphofructokinase activity, Isocitrate dehydrogenase activity, and Succinate dehydrogenase activity, n = 3. E-H) Heatmap of LC-MS data and quantitative analysis of the relative abundance of glycolysis and TCA cycle metabolites, n = 3. I) Schematic diagram of glycolysis and the TCA cycle. J) Representative CLSM images of RAW 264.7 cells treated with GMI gel for 72 h. K) Flow cytometry analysis of CD206 and CD86 expression. L, M) The secretion of IL-6 and TNF-α in the culture supernatant of RAW264.7 cells, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns, not significant (p > 0.05).

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: GMI gel -mediated metabolic reprogramming and its regulatory role in macrophage polarization. A-D) Hexokinase activity, Phosphofructokinase activity, Isocitrate dehydrogenase activity, and Succinate dehydrogenase activity, n = 3. E-H) Heatmap of LC-MS data and quantitative analysis of the relative abundance of glycolysis and TCA cycle metabolites, n = 3. I) Schematic diagram of glycolysis and the TCA cycle. J) Representative CLSM images of RAW 264.7 cells treated with GMI gel for 72 h. K) Flow cytometry analysis of CD206 and CD86 expression. L, M) The secretion of IL-6 and TNF-α in the culture supernatant of RAW264.7 cells, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns, not significant (p > 0.05).

Article Snippet: The expression levels of IL-6 and TNF-α in the culture medium were measured using commercial ELISA kits (Cusabio, Wuhan, China).

Techniques: Activity Assay, Liquid Chromatography with Mass Spectroscopy, Flow Cytometry, Expressing

GMI gel promotes the healing of infected pressure ulcers in vivo. A) Schematic diagram of GMI gel treatment of infected pressure ulcers. B) Photographs of wounds in mice at different treatment times. C) Signs of wound closure. D) Wound size at different treatment times, n = 3. E) H&E staining images of mouse wound tissue after different treatments on day 12. F) Masson staining images of mouse wound tissue after different treatments on day 12. G) Representative laser Doppler perfusion images of wounds in mice in each treatment group on day 12. H) Representative images of immunohistochemical staining for TNF- α, IL-6 and IL-10 12 days after treatment. I-L) Quantitative statistics of wound site blood perfusion, TNF- α, IL-6 and IL-10, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: GMI gel promotes the healing of infected pressure ulcers in vivo. A) Schematic diagram of GMI gel treatment of infected pressure ulcers. B) Photographs of wounds in mice at different treatment times. C) Signs of wound closure. D) Wound size at different treatment times, n = 3. E) H&E staining images of mouse wound tissue after different treatments on day 12. F) Masson staining images of mouse wound tissue after different treatments on day 12. G) Representative laser Doppler perfusion images of wounds in mice in each treatment group on day 12. H) Representative images of immunohistochemical staining for TNF- α, IL-6 and IL-10 12 days after treatment. I-L) Quantitative statistics of wound site blood perfusion, TNF- α, IL-6 and IL-10, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The expression levels of IL-6 and TNF-α in the culture medium were measured using commercial ELISA kits (Cusabio, Wuhan, China).

Techniques: Infection, In Vivo, Staining, Immunohistochemical staining