Journal: Frontiers in Immunology

Article Title: Xinfeng capsule attenuates ankylosing spondylitis by downregulating YTHDC1-mediated m 6 A modification of LINC01579 and suppressing IL-17/NF-κB signaling

doi: 10.3389/fimmu.2026.1762062

Figure Lengend Snippet: Pharmacology of XFC action on AS inflammation network. (A) XFC-AS-inflammation Wayne diagram; (B) XFC-AS-inflammation PPI network; (C) Identification of the top 10 targets of action based on the hubba plugin; (D) XFC component-AS-inflammation-pathway target network diagram; (E) CC bioprocesses; (F) KEGG analysis; (G) IL-17/NF-kB signaling pathway Schematic diagram. Pink squares in D are target genes, red arrows are potential pathways, and green circles are XFC core components.

Article Snippet: In vitro , co-culture experiments were performed using AS-FLSs and AS-PBMCs, with the IL-17 inhibitor Secukinumab (AIN457; Cat# HY-P9927, MCE; 16.2 ng/mL) employed to further elucidate pathway involvement.

Techniques:

Journal: Frontiers in Immunology

Article Title: Xinfeng capsule attenuates ankylosing spondylitis by downregulating YTHDC1-mediated m 6 A modification of LINC01579 and suppressing IL-17/NF-κB signaling

doi: 10.3389/fimmu.2026.1762062

Figure Lengend Snippet: Effects of AS-PBMCs and AS-FLS co-culture model on YTHDC1, LINC01579 and inflammatory cytokines. (A) MeRIP-qPCR for total m6A expression, (B) RT-qPCR for LINC01579 expression; (C) MeRIP-qPCR for LINC0159 m6A expression; (D) RT -qPCR to detect the expression of YTHDC1; (E) WB to detect the expression of YTHDC1; (F) ELISA to detect the expression of IL-6, IL-17 and TNF-a. All experiments were repeated three times. **p<0.01; ***p<0.001.

Article Snippet: In vitro , co-culture experiments were performed using AS-FLSs and AS-PBMCs, with the IL-17 inhibitor Secukinumab (AIN457; Cat# HY-P9927, MCE; 16.2 ng/mL) employed to further elucidate pathway involvement.

Techniques: Co-Culture Assay, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Xinfeng capsule attenuates ankylosing spondylitis by downregulating YTHDC1-mediated m 6 A modification of LINC01579 and suppressing IL-17/NF-κB signaling

doi: 10.3389/fimmu.2026.1762062

Figure Lengend Snippet: YTHDC1 Regulates LINC01579 Stability and Inflammatory Responses via the MUT1 m6A Site. (A–C) . ELISA to detect the expression of IL-6, IL-17 and TNF-a; (D) RT -qPCR to detect the expression of LINC0159; (E) Radiomycin D assay for LINC0157 stability; (F) WB to detect the expression of iIL-17A, IL-17RA, P-P65. All experiments were repeated three times. **p<0.01; ***p<0.001.

Article Snippet: In vitro , co-culture experiments were performed using AS-FLSs and AS-PBMCs, with the IL-17 inhibitor Secukinumab (AIN457; Cat# HY-P9927, MCE; 16.2 ng/mL) employed to further elucidate pathway involvement.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Xinfeng capsule attenuates ankylosing spondylitis by downregulating YTHDC1-mediated m 6 A modification of LINC01579 and suppressing IL-17/NF-κB signaling

doi: 10.3389/fimmu.2026.1762062

Figure Lengend Snippet: YTHDC1 regulates the activation of IL-17/NF-kB pathway by modulating the expression of LINC0157 m6A. (A) Nuclear-cytoplasmic fractionation assay to detect the expression of LINC0157; (B) Screening for the optimal small interfering RNA (siRNA) model for LINC0157; (C) RT-qPCR to detect the expression of LINC0157; (D) MeRIP-qPCR detection of LINC0159 m6A expression; (E) ELISA detection of IL-6 and TNF-a expression; (F) Colony formation assay to assess colony formation ability; (G) ELISA detection of IL-17A expression; (H) IF detection of p-P65 expression; (I) Screening for the optimal small interfering RNA model for YTHDC1; (J) RT-qPCR detection of YTHDC1 expression; (K) WB assay for YTHDC1 protein expression. (L) Radiomycin D assay for LINC0157 stability. (M) RT-qPCR assay for LINC0159 expression, (N) MeRIP-qPCR assay for LINC0159 m6A expression. All experiments were repeated three times. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: In vitro , co-culture experiments were performed using AS-FLSs and AS-PBMCs, with the IL-17 inhibitor Secukinumab (AIN457; Cat# HY-P9927, MCE; 16.2 ng/mL) employed to further elucidate pathway involvement.

Techniques: Activation Assay, Expressing, Fractionation, Small Interfering RNA, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Colony Assay

Journal: Frontiers in Immunology

Article Title: Xinfeng capsule attenuates ankylosing spondylitis by downregulating YTHDC1-mediated m 6 A modification of LINC01579 and suppressing IL-17/NF-κB signaling

doi: 10.3389/fimmu.2026.1762062

Figure Lengend Snippet: XFC Regulates LINC01579 and Downstream Inflammatory Responses via YTHDC1. (A) RT-qPCR assay to detect YTHDC1 expression, (B) RT-qPCR assay to detect LINC01579 expression, (C–E) . ELISA assay to detect IL-6, IL-17 and TNF-a expression. All experiments were repeated three times. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: In vitro , co-culture experiments were performed using AS-FLSs and AS-PBMCs, with the IL-17 inhibitor Secukinumab (AIN457; Cat# HY-P9927, MCE; 16.2 ng/mL) employed to further elucidate pathway involvement.

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Xinfeng capsule attenuates ankylosing spondylitis by downregulating YTHDC1-mediated m 6 A modification of LINC01579 and suppressing IL-17/NF-κB signaling

doi: 10.3389/fimmu.2026.1762062

Figure Lengend Snippet: XFC attenuates aberrant methylation and inflammatory response in PGIA mice (A) Study design and experimental schedule; (B) Representative hind paw observations of four groups of PGIA mice; (C) Representative images of Micro-CT scans of spinal joints in three groups, with osteogenesis and stenosis visible in PGIA mice (red arrows); (D) Mouse body weight; (E) Joint scores; (F) ELISA for ALT, AST, CRE (blood and kidney); (G) : ELISA for IL-6, IL-17 and TNF-a expression. (H) MeRIP-qPCR for total m6A expression. (I) RT-qPCR for LINC01579 expression; (J) RT- qPCR to detect the expression of YTHDC1; (K) WB to detect the expression of IL-17 and IL-17A. (L) HE staining of spondyloarthritic joints, PGIA mice were seen with inflammatory cell infiltration and synovial inflammation (black arrowheads), narrowing of joint space, cartilage degradation (green arrows), and bone erosion (yellow arrowheads). (M) Saffron O solid green staining of cartilage degradation was seen in PGIA mice (green arrows). All experiments were repeated three times. *p<0.05; **p<0.017

Article Snippet: In vitro , co-culture experiments were performed using AS-FLSs and AS-PBMCs, with the IL-17 inhibitor Secukinumab (AIN457; Cat# HY-P9927, MCE; 16.2 ng/mL) employed to further elucidate pathway involvement.

Techniques: Methylation, Micro-CT, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Staining

Journal: Frontiers in Immunology

Article Title: Xinfeng capsule attenuates ankylosing spondylitis by downregulating YTHDC1-mediated m 6 A modification of LINC01579 and suppressing IL-17/NF-κB signaling

doi: 10.3389/fimmu.2026.1762062

Figure Lengend Snippet: XFC improves AS inflammatory response by upregulating LINC01579 to inhibit IL-17/NF-kB activation (A) Representative hindpaws of five groups of PGIA mice; (B) Arthritis score; (C) ELISA detection of IL-6, IL-17, and TNF-a expression; (D) WB detection of IL-17 and IL-17A expression; (E) Spinal joint HE staining shows inflammatory cell infiltration and synovial inflammation (black arrows), narrowed joint spaces, cartilage degeneration (green arrows), and bone erosion (yellow arrows) in PGIA mice. (F) Fuchsin-O green staining, PGIA mice showed cartilage degeneration (green arrows). (G) RT-qPCR detection of LINC01579 expression in AS-FLS; (H) MeRIP-qPCR assay for LINC0159 m6A expression; (I) ELISA detection of IL-6, IL-17, and TNF-a expression in AS-FLS; (J) IF detection of p-P65 expression. All experiments were repeated three times. *p<0.05; **p<0.01.

Article Snippet: In vitro , co-culture experiments were performed using AS-FLSs and AS-PBMCs, with the IL-17 inhibitor Secukinumab (AIN457; Cat# HY-P9927, MCE; 16.2 ng/mL) employed to further elucidate pathway involvement.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Expressing, Staining, Quantitative RT-PCR

Journal: Discover Nano

Article Title: Chitosan nanoparticle encapsulated pentoxifylline improves renal protection and reduces oxidative stress in amikacin induced nephrotoxicity

doi: 10.1186/s11671-026-04515-8

Figure Lengend Snippet: Oxidative stress and inflammatory markers in kidney tissue homogenates of different experimental groups showing levels of MDA ( A ), SOD ( B ), GSH ( C ), IL-17 ( D ), TNF-α ( E ), IL-6 ( F ) and CRP ( G ). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. Significant differences between groups are indicated by horizontal brackets, with asterisks denoting significance levels ( p < 0.05). ns = not significant

Article Snippet: 1,1-Diphenyl-2-picrylhydrazyl (DPPH) (Sigma-Aldrich, USA) Amikacin (AMK) (Sigma-Aldrich, USA) BioTek Synergy H1 Microplate Reader (BioTek, USA) Bruker D8 Advance X-ray Diffractometer (Bruker, USA) Bruker Tensor II FTIR Spectrometer (Bruker, Germany) Chitosan (medium molecular weight, ≥75% deacetylated) (Sigma-Aldrich, USA) Creatinine Assay Kit (ab65340) (Abcam, USA) Glutathione (GSH) Assay Kit (MBS267424) (MyBioSource, USA) Hitachi SU3500 SEM (Hitachi, Japan) IL-17 ELISA Kit (E-EL-M0047) (Elabscience, USA) JEOL JEM-1400Flash TEM (JEOL, Japan) Malondialdehyde (MDA) Assay Kit (MBS741034) (MyBioSource, USA) Malvern Zetasizer Nano ZS90 (Malvern Panalytical, UK) Pentoxifylline (PTX) (Sigma-Aldrich, USA) Shimadzu UV-1800 Spectrophotometer (Shimadzu, Japan) Sodium Tripolyphosphate (TPP) (Sigma-Aldrich, USA) Superoxide Dismutase (SOD) Assay Kit (MBS2707323) (MyBioSource, USA) Sysmex CA-560 Coagulation Analyzer (Sysmex, Japan) Urea Assay Kit (ab83362) (Abcam, USA) Uric Acid Assay Kit (ab65344) (Abcam, USA)

Techniques:

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Aiolos and Eos drive distinct human TH17 functional states

doi: 10.1007/s00018-026-06089-1

Figure Lengend Snippet: Identification of Distinct Human TH17 Cell Subsets and Generation of Stable TH17 Clones from PBMC for Functional Characterization. A Schematic representation of the workflow to generate T H 17-IL22 + /IFNg + and T H 17-IL-10 + clones used to perform bulk ATAC-seq and RNA-seq data sets. In brief, peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood using density gradient centrifugation. The samples were enriched for CD4 + CCR6 + CXCR3- TH17 cells, referred to as “bulk TH17 cells.” Viable IL-17-producing cells were isolated by flow cytometry following a 3-hour stimulation with PMA and ionomycin using a IL-17 capture assay. The single TH17 cell clones were sorted into 384-well plates and expanded with allogeneic γ-irradiated feeder cells and phytohemagglutinin in complete medium containing IL-2. After approximately ten days, clones were transferred to 96-well plates for expansion, and following 2–3 weeks, their cytokine profiles were analyzed. T cell clones were then evaluated at two stages: day 0 (resting state) and day 5 (activated state). On day 5, they were stimulated for 48 hours with anti-CD3 and CD28, followed by an additional 3 days in uncoated plates. On both evaluation days, cells underwent further stimulation — 5 hours for protein analysis and 2 hours for RNA and chromatin-accessibility (ATAC-seq) analysis. Only TH17 clones exhibiting a stable cytokine profile after two rounds of resting and reactivation were selected for RNA-seq and ATAC-seq analysis. B Intracellular staining of IL-17 and IFNγ (top) and IL-22 and IL-10 (bottom) in a T H 17-IL10 + clone (right) and a T H 17-IL22 + /IFNg + clone (left) in the resting state (Day 0) and 5 days post-activation (Day 5). Numbers in quadrants indicate percent cells. C Frequency of IL-17+, IL-10+, IFNγ+, and IL-22+ cells among 6 independent TH17-IL-22 + /IFNγ + (left) and TH17-IL-10 + (right) clones at Day 0 and Day 5. Each symbol represents an individual T cell clone ( n = 6); data are shown as mean ± s.e.m. * P < 0.05, ** P < 0.01 (one-way ANOVA). TH17 clones were selected for RNA and ATAC-seq analysis based on the following criteria: ≥50% IL-17A+ cells at Day 0, ≥15% IL-22+ cells at Day 0 and Day 5, ≥15% IFNγ+ cells at Day 0 and Day 5 for TH17-IL-22 + /IFNγ + clones; ≥50% IL-17A+ cells at Day 0, ≥15% IL-10+ cells at Day 5 for TH17-IL-10 + clones

Article Snippet: Viable IL-17-producing bulk TH17 cells were sorted by flow cytometry using the IL-17 cytokine secretion assay (Miltenyi Biotec#130–094–536) following 3 h of stimulation with phorbol 12-myristate 13-acetate (PMA) (0.2 μM) and ionomycin (1 μg/ml) (both from Sigma-Aldrich), according to the manufacturer’s instructions.

Techniques: Clone Assay, Functional Assay, RNA Sequencing, Isolation, Gradient Centrifugation, Flow Cytometry, Irradiation, Staining, Activation Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Aiolos and Eos drive distinct human TH17 functional states

doi: 10.1007/s00018-026-06089-1

Figure Lengend Snippet: Identification of Distinct Human TH17 Cell Subsets and Generation of Stable TH17 Clones from PBMC for Functional Characterization. A Schematic representation of the workflow to generate T H 17-IL22 + /IFNg + and T H 17-IL-10 + clones used to perform bulk ATAC-seq and RNA-seq data sets. In brief, peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood using density gradient centrifugation. The samples were enriched for CD4 + CCR6 + CXCR3- TH17 cells, referred to as “bulk TH17 cells.” Viable IL-17-producing cells were isolated by flow cytometry following a 3-hour stimulation with PMA and ionomycin using a IL-17 capture assay. The single TH17 cell clones were sorted into 384-well plates and expanded with allogeneic γ-irradiated feeder cells and phytohemagglutinin in complete medium containing IL-2. After approximately ten days, clones were transferred to 96-well plates for expansion, and following 2–3 weeks, their cytokine profiles were analyzed. T cell clones were then evaluated at two stages: day 0 (resting state) and day 5 (activated state). On day 5, they were stimulated for 48 hours with anti-CD3 and CD28, followed by an additional 3 days in uncoated plates. On both evaluation days, cells underwent further stimulation — 5 hours for protein analysis and 2 hours for RNA and chromatin-accessibility (ATAC-seq) analysis. Only TH17 clones exhibiting a stable cytokine profile after two rounds of resting and reactivation were selected for RNA-seq and ATAC-seq analysis. B Intracellular staining of IL-17 and IFNγ (top) and IL-22 and IL-10 (bottom) in a T H 17-IL10 + clone (right) and a T H 17-IL22 + /IFNg + clone (left) in the resting state (Day 0) and 5 days post-activation (Day 5). Numbers in quadrants indicate percent cells. C Frequency of IL-17+, IL-10+, IFNγ+, and IL-22+ cells among 6 independent TH17-IL-22 + /IFNγ + (left) and TH17-IL-10 + (right) clones at Day 0 and Day 5. Each symbol represents an individual T cell clone ( n = 6); data are shown as mean ± s.e.m. * P < 0.05, ** P < 0.01 (one-way ANOVA). TH17 clones were selected for RNA and ATAC-seq analysis based on the following criteria: ≥50% IL-17A+ cells at Day 0, ≥15% IL-22+ cells at Day 0 and Day 5, ≥15% IFNγ+ cells at Day 0 and Day 5 for TH17-IL-22 + /IFNγ + clones; ≥50% IL-17A+ cells at Day 0, ≥15% IL-10+ cells at Day 5 for TH17-IL-10 + clones

Article Snippet: PE IL-17 Secretion Assay- detection kit , Miltenyi , 130-094-536.

Techniques: Clone Assay, Functional Assay, RNA Sequencing, Isolation, Gradient Centrifugation, Flow Cytometry, Irradiation, Staining, Activation Assay