Journal: Journal of Translational Autoimmunity

Article Title: T cell proliferative response to a homocitrullinated peptide correlates with joint pathology in collagen induced arthritis

doi: 10.1016/j.jtauto.2025.100345

Figure Lengend Snippet: RA-Specific Serum IgG Antibodies in DBA/1J Mice. Sera were collected biweekly from DBA/1J mice immunized with bovine type II collagen (bCII) (arthritic: N = 9–18; non-arthritic: N = 17–37), or PBS (N = 14) and tested via indirect ELISA for A) anti-CitP, and B) anti-HomoCitP IgG antibodies. Graphs show the median [IQR] for each group, with a cut-off indicating the limit of detection (dashed line). Statistical analysis was performed using a mixed effects model with the Geisser-Greenhouse correction and Tukey's multiple comparisons test. The resulting p-values are indicated on the graphs.

Article Snippet: The anti-mCII ELISA kits were used according to manufacturer's protocol (2036T; Chondrex).

Techniques: Indirect ELISA

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: Significant Ups genes in the acetabular cartilage of the rats with DDH, as determined by mRNA-seq analysis. (A) Neonatal rats were subjected to straight-leg swaddling for 10 days. A total of 4 weeks later, (a) morphology, (b) H&E staining and (c) SO/FG staining were assessed in the hip joints of the rats. (B) Immunohistochemical staining and (E) semi-quantitative analysis of COL2A1 in the acetabular cartilage of the 4-week-old control rats and rats with DDH. (C) Relative gene expression levels of COL2A1, IL-1β and IL-6 in the acetabular cartilage of the 4-week-old control rats and rats with DDH, as determined by reverse transcription-quantitative PCR. (D) Levels of IL-6 in the supernatants of acetabular cartilage from the 4-week-old rats with DDH, as determined by ELISA. (F) Significant Ups in the acetabular cartilage of rats with DDH were identified by mRNA-seq analysis with the screening criteria of log 2 FC >4 and P.adj<0.01. Principal component analysis results and a heatmap of these significant Ups are shown. (G) GO enrichment analysis of significant Ups in the acetabular cartilage of the rats with DDH. Data are presented as the mean ± SD. *** P<0.001. (C-E) Representative results of four independent experiments are shown. Ups, upregulated genes; BP, biological process; CC, cellular component; COL2A1, type II collagen; DDH, developmental dysplasia of the hip; FC, fold change; GO, Gene Ontology; H&E, hematoxylin and eosin; IHC, immunohistochemistry; IL, interleukin; IOD, integrated optical density; MF, molecular function; mRNA-seq, mRNA sequencing; SO/FG, Safranin O/Fast Green.

Article Snippet: Protein concentrations were measured using a BCA assay kit (cat. no. PK10026; Wuhan Sanying Biotechnology), and the levels of TNF-α and IL-6 in the tissue homogenates and cell supernatants were measured using the Rat TNF-α ELISA Kit (cat. no. EK382; Multi Sciences Biotech) and Rat IL-6 ELISA Kit (cat. no. EK306; Multi Sciences Biotech), according to the manufacturers' instructions.

Techniques: Staining, Immunohistochemical staining, Control, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Sequencing

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: DUSP26 overexpression alleviates IL-1β-induced chondrocyte injury. Rat DUSP26 overexpression adenovirus was constructed and used to infect chondrocytes. After 24 h, the chondrocytes were subjected to 10 ng/ml IL-1β. (A) Expression levels of COL2A1 and COL1A1 in the chondrocytes were detected by RT-qPCR and western blotting. (B) Relative mRNA expression levels of TNF-α and IL-6 in the chondrocytes were determined by RT-qPCR. (C) Levels of TNF-α and IL-6 in the supernatants of the chondrocytes were determined by ELISA. Two adenovirus-mediated shRNAs against rat DUSP26 were constructed and used to infect chondrocytes. After 24 h, the chondrocytes were subjected to 10 ng/ml IL-1β. (D) Expression levels of COL2A1 and COL1A1 in the chondrocytes were evaluated by RT-qPCR and western blotting. (E) Relative mRNA expression levels of TNF-α and IL-6 in the chondrocytes were determined by RT-qPCR. (F) Levels of TNF-α and IL-6 in the supernatants of the chondrocytes were detected by ELISA. Data are presented as the mean ± SD. * P<0.05, ** P<0.01 and *** P<0.001. Representative results of three independent experiments are shown. COL1A1, type I collagen; COL2A1, type II collagen; EP, empty adenoviral vector; DUSP26, dual-specificity phosphatase 26; IL, interleukin; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: Protein concentrations were measured using a BCA assay kit (cat. no. PK10026; Wuhan Sanying Biotechnology), and the levels of TNF-α and IL-6 in the tissue homogenates and cell supernatants were measured using the Rat TNF-α ELISA Kit (cat. no. EK382; Multi Sciences Biotech) and Rat IL-6 ELISA Kit (cat. no. EK306; Multi Sciences Biotech), according to the manufacturers' instructions.

Techniques: Over Expression, Construct, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Integrative Medicine Research

Article Title: Therapeutic effects of pomegranate hot-water extract via inhibition of apoptosis and oxidative stress in a DHEA-induced mouse model of PCOS

doi: 10.1016/j.imr.2025.101264

Figure Lengend Snippet: Pg-hWE alleviates PCOS symptoms in a mouse model based on H&E staining. (A) PCOS mouse model construction. The mice were subcutaneously administered with 30 mg/kg of DHEA for six weeks. One week prior to start of SC administration, each group was administered the appropriate drug (tap water, dextrin, and Pg-hWE) orally. (B) Body weight was measured once a week. (C) Results of the AMH ELISA with mouse serum. (D) Mouse ovary histology by H&E staining. (E) Results of ovarian follicles by type. # p < 0.05 ## p < 0.01 compared with the Normal group, * p < 0.05 and ⁎⁎ p < 0.01 compared with the PCOS group.

Article Snippet: ELISA of mouse serum was performed using the CUSABIO's anti-Müllerian hormone ELISA kit (CUSABIO, Houston, TX, USA) as reported previously, and all preparation and experimental procedures were performed according to the protocol provided by CUSABIO.

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Bioengineered extracellular vesicles escape lysosomal degradation and deliver Tet-PKM2 for macrophage immunometabolic reprogramming and periodontitis treatment

doi: 10.1016/j.bioactmat.2026.01.002

Figure Lengend Snippet: Immunomodulatory effects of the bioengineered LEVs Tet−PKM2 @TA in terms of their ability to modulate macrophage polarization in vitro . The macrophages were treated with 100 ng/mL LPS for 24 h and then treated with PBS (Control), 100 μg/mL LEVs PKM2 , LEVs Tet−PKM2 , or LEVs Tet−PKM2 @TA for another 24 h. ( A ) The relative mRNA expression levels of M1 polarization-related genes ( IL-6 and IL-1β ) and M2 polarization-related genes ( IL-4 and Arg-1 ) in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups (qRT‒PCR) ( n = 3). ( B ) Concentrations of M1-related cytokines (IL-6 and TNF-α) and M2-related cytokines (IL-4 and IL-10) in the supernatants of the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups (ELISA) ( n = 3). ( C ) Representative immunofluorescence images and quantification of the expression levels of M1-related proteins (iNOS and CCR7) and M2-related proteins (CD163, CD206, and Arg-1) in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 3). The data are expressed as the mean ± SEM. Statistical analysis was performed with one-way ANOVA ( A , B , and C ). ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 indicate significant differences between the indicated columns.

Article Snippet: Kits were sourced as follows: TNF-α, IL-4, and IL-10 from Fankew (Shanghai Kexing Trading Co., Ltd., China) and IL-6 from Proteintech.

Techniques: In Vitro, Control, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Materials Today Bio

Article Title: One-step green assembly of natural polyphenol-based nobiletin nanoparticles for ulcerative colitis therapy

doi: 10.1016/j.mtbio.2026.102887

Figure Lengend Snippet: | In vivo anti-oxidant and anti-inflammatory effects of NTAl NPs against UC. A) H&E-stained sections of mouse colon following treatment with indicated materials. B) H&E staining histological scores. C) Typical TEM images showing the microstructures of colonic epitheliums. D-F) the expression of oxidative stress markers including MDA(D), SOD (E), and GSH-Px (F). G-J) the expression of colonic inflammatory cytokines including pro-inflammatory cytokines TNF-α (G), IL-1β (H), IL-6 (I) and anti-inflammatory cytokines IL-10 (J). All data are shown as mean ± s.d; n = 6. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Commercial ELISA kits including Mouse TNF-α (EK282, Multi Sciences), Mouse IL-1β (EK201B, Multi Sciences), Mouse IL-6 (EK206, Multi Sciences), and Mouse IL-10 (EK210, Multi Sciences) were used to quantify pro-inflammatory cytokines and the anti-inflammatory cytokine according to the manufacturer's instructions.

Techniques: In Vivo, Staining, Expressing

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: Significant Ups genes in the acetabular cartilage of the rats with DDH, as determined by mRNA-seq analysis. (A) Neonatal rats were subjected to straight-leg swaddling for 10 days. A total of 4 weeks later, (a) morphology, (b) H&E staining and (c) SO/FG staining were assessed in the hip joints of the rats. (B) Immunohistochemical staining and (E) semi-quantitative analysis of COL2A1 in the acetabular cartilage of the 4-week-old control rats and rats with DDH. (C) Relative gene expression levels of COL2A1, IL-1β and IL-6 in the acetabular cartilage of the 4-week-old control rats and rats with DDH, as determined by reverse transcription-quantitative PCR. (D) Levels of IL-6 in the supernatants of acetabular cartilage from the 4-week-old rats with DDH, as determined by ELISA. (F) Significant Ups in the acetabular cartilage of rats with DDH were identified by mRNA-seq analysis with the screening criteria of log 2 FC >4 and P.adj<0.01. Principal component analysis results and a heatmap of these significant Ups are shown. (G) GO enrichment analysis of significant Ups in the acetabular cartilage of the rats with DDH. Data are presented as the mean ± SD. *** P<0.001. (C-E) Representative results of four independent experiments are shown. Ups, upregulated genes; BP, biological process; CC, cellular component; COL2A1, type II collagen; DDH, developmental dysplasia of the hip; FC, fold change; GO, Gene Ontology; H&E, hematoxylin and eosin; IHC, immunohistochemistry; IL, interleukin; IOD, integrated optical density; MF, molecular function; mRNA-seq, mRNA sequencing; SO/FG, Safranin O/Fast Green.

Article Snippet: Protein concentrations were measured using a BCA assay kit (cat. no. PK10026; Wuhan Sanying Biotechnology), and the levels of TNF-α and IL-6 in the tissue homogenates and cell supernatants were measured using the Rat TNF-α ELISA Kit (cat. no. EK382; Multi Sciences Biotech) and Rat IL-6 ELISA Kit (cat. no. EK306; Multi Sciences Biotech), according to the manufacturers' instructions.

Techniques: Staining, Immunohistochemical staining, Control, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Sequencing

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: DUSP26 overexpression alleviates IL-1β-induced chondrocyte injury. Rat DUSP26 overexpression adenovirus was constructed and used to infect chondrocytes. After 24 h, the chondrocytes were subjected to 10 ng/ml IL-1β. (A) Expression levels of COL2A1 and COL1A1 in the chondrocytes were detected by RT-qPCR and western blotting. (B) Relative mRNA expression levels of TNF-α and IL-6 in the chondrocytes were determined by RT-qPCR. (C) Levels of TNF-α and IL-6 in the supernatants of the chondrocytes were determined by ELISA. Two adenovirus-mediated shRNAs against rat DUSP26 were constructed and used to infect chondrocytes. After 24 h, the chondrocytes were subjected to 10 ng/ml IL-1β. (D) Expression levels of COL2A1 and COL1A1 in the chondrocytes were evaluated by RT-qPCR and western blotting. (E) Relative mRNA expression levels of TNF-α and IL-6 in the chondrocytes were determined by RT-qPCR. (F) Levels of TNF-α and IL-6 in the supernatants of the chondrocytes were detected by ELISA. Data are presented as the mean ± SD. * P<0.05, ** P<0.01 and *** P<0.001. Representative results of three independent experiments are shown. COL1A1, type I collagen; COL2A1, type II collagen; EP, empty adenoviral vector; DUSP26, dual-specificity phosphatase 26; IL, interleukin; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: Protein concentrations were measured using a BCA assay kit (cat. no. PK10026; Wuhan Sanying Biotechnology), and the levels of TNF-α and IL-6 in the tissue homogenates and cell supernatants were measured using the Rat TNF-α ELISA Kit (cat. no. EK382; Multi Sciences Biotech) and Rat IL-6 ELISA Kit (cat. no. EK306; Multi Sciences Biotech), according to the manufacturers' instructions.

Techniques: Over Expression, Construct, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: Inactivation of HDAC1/2/8 inhibits DUSP26 silencing-triggered cartilage degeneration. (A) Total protein and phosphorylation levels of HDAC1, HDAC2 and HDAC8 in IL-1β-treated chondrocytes determined by western blotting. (B) Relative mRNA expression levels of HDAC1, HDAC2 and HDAC8 in chondrocytes, as determined by reverse transcription-quantitative PCR. (C) Relative mRNA expression levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after knocking down HDAC1/2/8. (D) Relative mRNA expression levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after intervention with the HDAC inhibitor TSA. (E) Protein levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after knocking down HDAC1/2/8. (F) Protein levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after intervention with the HDAC inhibitor TSA. Data are presented as the mean ± SD. * P<0.05, ** P<0.01 and *** P<0.001. (B-D) Representative results of three independent experiments are shown. COL1A1, type I collagen; DUSP26, dual-specificity phosphatase 26; HDAC, histone deacetylase; IL, interleukin; NC, negative control; ns, no significance; sh, short hairpin; TSA, trichostatin A.

Article Snippet: Protein concentrations were measured using a BCA assay kit (cat. no. PK10026; Wuhan Sanying Biotechnology), and the levels of TNF-α and IL-6 in the tissue homogenates and cell supernatants were measured using the Rat TNF-α ELISA Kit (cat. no. EK382; Multi Sciences Biotech) and Rat IL-6 ELISA Kit (cat. no. EK306; Multi Sciences Biotech), according to the manufacturers' instructions.

Techniques: Phospho-proteomics, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Histone Deacetylase Assay, Negative Control

Journal: Materials Today Bio

Article Title: One-step green assembly of natural polyphenol-based nobiletin nanoparticles for ulcerative colitis therapy

doi: 10.1016/j.mtbio.2026.102887

Figure Lengend Snippet: | In vivo anti-oxidant and anti-inflammatory effects of NTAl NPs against UC. A) H&E-stained sections of mouse colon following treatment with indicated materials. B) H&E staining histological scores. C) Typical TEM images showing the microstructures of colonic epitheliums. D-F) the expression of oxidative stress markers including MDA(D), SOD (E), and GSH-Px (F). G-J) the expression of colonic inflammatory cytokines including pro-inflammatory cytokines TNF-α (G), IL-1β (H), IL-6 (I) and anti-inflammatory cytokines IL-10 (J). All data are shown as mean ± s.d; n = 6. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Commercial ELISA kits including Mouse TNF-α (EK282, Multi Sciences), Mouse IL-1β (EK201B, Multi Sciences), Mouse IL-6 (EK206, Multi Sciences), and Mouse IL-10 (EK210, Multi Sciences) were used to quantify pro-inflammatory cytokines and the anti-inflammatory cytokine according to the manufacturer's instructions.

Techniques: In Vivo, Staining, Expressing