Journal: Respiratory Research

Article Title: Plasma proteomic and machine learning models for differentiating idiopathic pulmonary fibrosis and connective tissue disease–associated interstitial lung disease: findings from a prospective cohort

doi: 10.1186/s12931-026-03596-4

Figure Lengend Snippet: Independent protein-level validation of candidate biomarkers in CTD-ILD and IPF. A Plasma levels of TWEAK, ( B ) MMP-10, and ( C ) ADA were quantified by ELISA in an independent validation cohort (CTD-ILD, n = 38; IPF, n = 22). Statistical significance was assessed using the Mann–Whitney U test (*** p < 0.001)

Article Snippet: FGF-19 concentrations were determined using the Human FGF-19 ELISA kit (DY969, R&D Systems), and TWEAK levels were quantified using the Human TWEAK ELISA kit (DY1090, R&D Systems).

Techniques: Biomarker Discovery, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Advanced Science

Article Title: Reprogramming of Fatty Acid Metabolism via PPARα‐Orchestrated FADS2 in Keratinocytes Modulates Skin Inflammation in Psoriasis

doi: 10.1002/advs.202417049

Figure Lengend Snippet: FADS2 modulates psoriatic inflammation in keratinocytes through NF‐κB activation. A) RT‐qPCR analysis of the indicated genes in HaCaT cells transfected with FADS2 siRNA (si FADS2 ) or control siRNA (siNC) for 24 h, followed by stimulation with PBS or a cytokine cocktail (M5) for 12 h (n = 3). B) ELISA quantification of CXCL1 and CXCL8 protein levels in both cell lysates and supernatants of HaCaT cells treated as (A) (n = 3). C) Top 10 enriched Gene Ontology (GO) molecular function terms from RNA‐sequencing (RNA‐seq) analysis of differential expression genes (DEGs) in FADS2 ‐silenced (si FADS2 ) versus control (siNC) HaCaT cells after M5 stimulation. DEGs were defined by |fold change| >1.5 & adjusted P <0.05. D) Gene set enrichment analysis (GSEA) of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment from RNA‐seq results in FADS2 ‐silenced (si FADS2 ) versus control (siNC) HaCaT cells after M5 stimulation. E) Representative immunofluorescence images of phosphorylated NF‐κB p65 (pNF‐κB) and K14 co‐staining in normal skin from healthy controls and lesional skin tissue from psoriasis patients. F) Immunoblotting of pNF‐κB and total‐NF‐κB p65 (NF‐κB) in HaCaT cells transfected with si FADS2 or siNC for 24 h and stimulated with PBS or M5 for 1 h. G) RT‐qPCR analysis of the indicated genes in HaCaT cells transfected with si FADS2 or siNC and stimulated with M5 after BAY 11–7082 or DMSO pretreatment (n = 3). H) RT‐qPCR analysis of FADS2 and the indicated genes in HaCaT cells transfected with FADS2 overexpression plasmids ( FADS2 OE) or empty vector for 48 h followed by PBS or M5 treatment for 10 h (n = 3). Scale bar, 50 µm. Data are presented as mean ± SD. Statistical significance was determined by two‐way ANOVA (A,G,H) or unpaired two‐tailed Student's t ‐test (B, H). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: Similarly, human CXCL1 (#70‐EK196), CSF3 (#70‐EK169), and CXCL8 (#70‐EK108/2) levels in cultured human keratinocyte lysates and supernatants were measured using corresponding human ELISA kits (Lianke Bio, China) according to the provided protocols.

Techniques: Activation Assay, Quantitative RT-PCR, Transfection, Control, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Quantitative Proteomics, Immunofluorescence, Staining, Western Blot, Over Expression, Plasmid Preparation, Two Tailed Test

Journal: Advanced Science

Article Title: Reprogramming of Fatty Acid Metabolism via PPARα‐Orchestrated FADS2 in Keratinocytes Modulates Skin Inflammation in Psoriasis

doi: 10.1002/advs.202417049

Figure Lengend Snippet: PPARα acts as an upstream positive regulator of FADS2 in psoriatic keratinocytes. A) RT‐qPCR analysis of PPARA expression in normal skin from healthy controls (NN, n = 4) and lesional skin tissue from psoriasis patients (PS, n = 5). B) Representative immunofluorescence images of PPARα and K14 co‐staining in normal skin from healthy controls and lesional skin tissue from psoriasis patients. C) RT‐qPCR analysis of PPARA in psoriatic lesional skin before and 10 weeks after infliximab treatment (n = 5). D) Representative images of PPARα immunofluorescence staining in lesional skin from psoriasis patients at baseline and week 10 following infliximab treatment. E) Representative immunofluorescence images of PPARα staining in healthy skin from control mice and skin lesions from IMQ‐induced psoriasis mouse model. F,G) RT‐qPCR analysis of PPARA (F) and FADS2 (G) expression in HaCaT cells stimulated with M5 for 12 h (n = 3). H) Immunoblotting of PPARα and FADS2 in HaCaT cells stimulated with M5 cytokines for the indicated time. I) RT‐qPCR analysis of PPARA , FADS2 , and indicated genes in HaCaT cells transfected with PPARA siRNA (si PPARA ) and control siRNA (siNC) for 24 h, followed by PBS or M5 stimulation for 12 h (n = 3). J) RT‐qPCR analysis of FADS2 and indicated genes in HaCaT cells pretreated with WY14643 or DMSO for 21 h, followed by PBS or M5 stimulation for 3 h (n = 3). K) ELISA quantification of CXCL1, CXCL8, and CSF3 protein levels in cell lysates and supernatants from HaCaT cells treated as in (J) (n = 3). L) RT‐qPCR analysis of the indicated genes in HaCaT cells transfected with si FADS2 and siNC for 24 h, followed by M5 stimulation for 3 h before WY14643 or DMSO pretreatment (n = 3). Scale bar, 50 µm. Data are presented as mean ± SD. Statistical significance was determined by unpaired two‐tailed Student's t test (A,F,G,K), paired two‐tailed Student's t test (C), or one‐way ANOVA (I,J,L). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: Similarly, human CXCL1 (#70‐EK196), CSF3 (#70‐EK169), and CXCL8 (#70‐EK108/2) levels in cultured human keratinocyte lysates and supernatants were measured using corresponding human ELISA kits (Lianke Bio, China) according to the provided protocols.

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Control, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay, Two Tailed Test