Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Schematic representation of the human collagen XVIII variants, termed as SHORT, MIDDLE and LONG/FZ. Collagenous sequences are shown in white. Non-collagenous (NC) amino terminal sequences common to all variants are shown in black. Non-collagenous amino terminal sequences common to the two long variants are shown in grey. A non-collagenous amino terminal sequence specific to the LONG/FZ variant is marked with waves. The amino acid lengths (aa) and the predicted molecular masses (kDa) of these sequences are given, as are those of the full length type XVIII collagen molecules. Signal sequences are indicated by vertical and horizontal hatching. The epitopes of the antibodies used in this work are shown by black horizontal lines. The molecular masses of some endostatin-containing carboxy-terminal fragments are also indicated.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Sequencing, Variant Assay

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Immunoprecipitation with anti-ALL antibody and detection with anti-LONG huXVIII antibody. Elevated type XVIII collagen precursors in the plasma of ALI patients compared with normal controls is shown. ALI = acute lung injury.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Immunoprecipitation, Clinical Proteomics

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Western blot by anti-ALL huXVIII antibody. It shows elevated type XVIII collagen precursors in the bronchoalveolar lavage fluid (BALF) of patients with acute lung injury compared with normal controls. ALI = acute lung injury.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Western Blot

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: GA attenuates oxidative stress by inhibiting inflammation to alleviate tendinopathy. In vivo, Wistar rats were divided into 6 groups: vehicle, ciprofloxacin, ciprofloxacin + 25 mg/kg GA, ciprofloxacin + 50 mg/kg GA, ciprofloxacin + 100 mg/kg GA and positive control (five rats in each group). ( A ) H&E and Masson staining were used to observe collagen organization (blue: collagen fibres; red: muscle fibres); Sirius red staining was used to distinguish between type I and type III collagen (scale bars: 10×, 100 μm); ( B ) The expression of collagen I/III was determined by immunofluorescence staining (scale bars: 10×, 100 μm); ( C ) The protein expression of PTGS2 in rat tendon tissues was assessed by immunohistochemical staining (scale bars: 10×, 100 μm). ( D-E ) Biological process and protein function enrichment analysis of the TMT proteomics data. ( F-H ) The serum levels of MDA, GSH and IL-1β were determined with the relevant kits; ( n = 5, data are presented as mean ± SD, ** p < 0.01, *** p < 0.001 vs. vehicle group; ## p < 0.01, ### p < 0.001 vs. Ciprofloxacin group)

Article Snippet: The anti-NLRP3 antibody (#BA3677) and anti-Collagen Type III antibody (#M00788-1) were obtained from Boster, and the anti-ASC antibody (#HA721306) was obtained from Hangzhou Huaan Biotechnology.

Techniques: In Vivo, Positive Control, Staining, Expressing, Immunofluorescence, Immunohistochemical staining

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: GA increases TSC viability by inhibiting oxidative stress and inflammatory responses. In vitro, TSCs were pretreated with different concentrations of GA for 12 h and then subjected to the prescribed concentration of H 2 O 2 (100 µM) for 24 h. ( A-B ) Cell viability was assessed by MTT assay; ( C ) Cell viability was assessed by CCK-8 assay; ( D-E ) The levels of GSH and MDA were determined with the relevant kits; ( F ) The content of ROS in TSCs was measured by flow cytometry using DCFH-DA probe; ( G-H ) Western blotting and qPCR were used to assess the protein and gene expression of inflammatory cytokines, including IL-1β, IL-6, and IL-8; ( I ) Tenomodulin, Collagen I and Collagen III protein levels were assessed via western blotting. ( n = 3, data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H₂O₂ group)

Article Snippet: The anti-NLRP3 antibody (#BA3677) and anti-Collagen Type III antibody (#M00788-1) were obtained from Boster, and the anti-ASC antibody (#HA721306) was obtained from Hangzhou Huaan Biotechnology.

Techniques: In Vitro, Concentration Assay, MTT Assay, CCK-8 Assay, Flow Cytometry, Western Blot, Gene Expression, Control

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: GA protects TSCs against oxidative stress via the HMGB1-cGAS-NLRP3 axis in tendinopathy. Wistar rats were divided into five groups (five rats per group): vehicle, ciprofloxacin, ciprofloxacin + 50 mg/kg GA, ciprofloxacin + 50 mg/kg GA + VA-Lip-Ctrl-Plasmid, and ciprofloxacin + 50 mg/kg GA + VA-Lip-cGAS-Plasmid. ( A-B ) Immunohistochemical detection for cGAS, HMGB1, NLRP3 and ASC in tendon tissues (scale bars: 10×, 100 μm); ( C ) H&E and Masson staining were used to observe the collagen organization of tendon tissues (blue indicates collagen fibres and red indicates muscle fibres); Sirius red staining was used to distinguish between type I and type III collagen (Scale bars: 10×, 100 μm); ( D ) The expression of collagen I/III were assessed by immunofluorescence staining (scale bars: 10×, 100 μm); ( E ) Immunohistochemical detection of the expression of NLRP3 and ASC (scale bars: 10×, 100 μm); ( F-H ) Serum levels of MDA and IL-1β were measured by commercial ELISA kits; ( I ) Immunofluorescence staining was used to detect the expression and colocalization of cGAS and HMGB1 in the tendon tissues of the rats (scale bars: 20×, 50 μm). ( J ) Immunofluorescence staining was used to detect the colocalization of cGAS and HMGB1 in the tendon tissue of clinical sample 1. ( n = 5, data are presented as mean ± SD, ** p < 0.01, *** p < 0.001 vs. vehicle group; ## p < 0.01, ### p < 0.001 vs. Ciprofloxacin group; && p < 0.01, &&& p < 0.001 vs. Ciprofloxacin + GA group)

Article Snippet: The anti-NLRP3 antibody (#BA3677) and anti-Collagen Type III antibody (#M00788-1) were obtained from Boster, and the anti-ASC antibody (#HA721306) was obtained from Hangzhou Huaan Biotechnology.

Techniques: Plasmid Preparation, Immunohistochemical staining, Staining, Expressing, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: The Journal of pathology

Article Title: A 3D tri-culture system reveals that activin receptor-like kinase 5 and connective tissue growth factor drive human glomerulosclerosis

doi: 10.1002/path.4960

Figure Lengend Snippet: (A) MC 3D monoculture without the addition of TGFβ, and (B) in the presence of TGFβ, shown by H&E staining of paraffin-embedded sections. Human collagen type I/III (immunostained with an antibody that recognises both collagen type I and III) (C) and human collagen type IV (D) can be demonstrated in nodules by immunoperoxidase staining. (E) High-power view of MC nodule by scanning electron microscopy in cross section. (F) MC nodule count and collagen type I alpha1 (COL1a1), and collagen type IV alpha1 (COL4a1) RNA quantification (n=3). (G) Effect of ALK5 inhibition (Alk5i) on MC nodule formation (n=3). (H) Effect of SMAD2 or SMAD3 siRNA knockdown in MCs on nodule formation (n=3) with immunoblots demonstrating degree of knockdown. [DharmaFECT (DH), non-targeting siRNA control (siCP), siSMAD2 (siS2), siSMAD3 (siS3), total SMAD2 (tSMAD2), total SMAD3 (tSMAD3)]. Error bars represent 1 s.d. NS, not significant; ***, p<0.001; **, p<0.01: 2-tailed Student’s t test.

Article Snippet: After culture, matrices were embedded in HistoGel, fixed in 4% paraformaldehyde and paraffin wax-embedded for sectioning and staining for human collagen type IV (mouse anti-human collagen IV monoclonal antibody 2150–0121, Bio-Rad, CA, USA) and human collagen type I/III (rabbit anti-human collagen I/III polyclonal antibody 2150–2210, Bio-Rad, CA, USA).

Techniques: Staining, Immunoperoxidase Staining, Electron Microscopy, Inhibition, Western Blot