Journal: International Journal of Molecular Medicine

Article Title: Orcinol glucoside ameliorates pulmonary fibrosis by suppressing hyaluronic acid synthesis and macrophage M2 polarization via targeting hyaluronic acid synthase 2

doi: 10.3892/ijmm.2026.5764

Figure Lengend Snippet: Elevated HAS2 expression and HA accumulation in murine and cellular fibrosis models. (A) Schematic of the experimental design for the time-course study. C57BL/6J mice received a single intratracheal dose of BLM or saline and were sacrificed at the indicated time points (days 0, 3, 5, 7, 10, 14 and 21) for sample collection (n=5 for per group). (B) Representative western blot images (left panel) and densitometric quantification (right panel) showing the protein expression levels of COL1A1 and HAS2 in lung tissues across the time course. β-tubulin served as the loading control (n=4 for per group). (C) ELISA quantification of HA levels in BALF at different days post-BLM injury (n=5 for per group). (D) Schematic diagram illustrating the workflow for the isolation and culture of primary mouse lung fibroblasts from BLM-treated fibrotic mice. (E) Western blotting analysis of HAS2 and COL1A1 protein expression in primary lung fibroblasts isolated from control (saline) and BLM-induced fibrotic mice. Representative blot images (left panel) and densitometric quantification of protein levels normalized to β-tubulin are shown (n=3 for each group). (F) Schematic diagram depicting TGF-β1-induced transition of NIH/3T3 fibroblasts into myofibroblasts. (G) COL1A1, ACTA2, and HAS2 mRNA expression in NIH/3T3 cells was detected using PCR after 5 ng/ml TGF-β1 administration for 12 h (n=3-4 for each group). (H) HAS2 protein expression in NIH/3T3 was detected using western blotting after 5 ng/ml TGF-β1 administration for 24 h (n=3 for each group). (I) ELISA was used to quantify HA concentrations in the culture media of myofibroblasts (n=6 for each group). * P<0.05, ** P<0.01, *** P<0.001. HAS2, hyaluronic acid synthase 2; HA, hyaluronic acid; BLM, bleomycin; BALF, bronchoalveolar lavage fluid; COL1A1, Collagen type I α 1 chain; ACTA2, actin alpha 2, smooth muscle.

Article Snippet: Antibodies for Collagen type I α 1 chain (COL1A1; cat. no. 72026; 1:1,000), Phospho-STAT6 (Tyr641; cat. no. 56554S; 1:1,000) and α-smooth muscle actin (α-SMA; cat. no. 19245; 1:1,000) were obtained from Cell Signaling Technology, Inc.

Techniques: Expressing, Saline, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Isolation

Journal: International Journal of Molecular Medicine

Article Title: Orcinol glucoside ameliorates pulmonary fibrosis by suppressing hyaluronic acid synthesis and macrophage M2 polarization via targeting hyaluronic acid synthase 2

doi: 10.3892/ijmm.2026.5764

Figure Lengend Snippet: OG exerts an anti-pulmonary fibrosis effect by impairing myofibroblast-mediated macrophage M2 polarization. (A) Schematic workflow: Fibroblast treatment with OG during TGF-β1 stimulation, followed by conditioned medium collection and macrophage culture for downstream assays. (B) TGF-β1 release from macrophages measured by ELISA (n=3 for each group). (C) Flow cytometric analysis of CD206 and CD86 expression in macrophages cultured in conditioned medium. The bar graph quantifies the proportion of CD206 + CD86 − cells (M2-like macrophage subpopulation; n=3 for each group). (D) Animal experimental protocol. (E) A BLM-induced mouse model was established to assess the anti-fibrotic effects of OG. Histopathological alterations in lung tissues from different groups were assessed using H&E and Masson's trichrome staining. Scale bars: 500 μ m; magnification, ×40. (F) Western blotting analysis to measure the expression levels of fibrotic markers, COL1A1 and α-SMA, in lung tissues (n=4 for each group). (G) The levels of HA in mouse serum were quantified by ELISA. (H) The levels of TGF-β1 in mouse serum were quantified by ELISA (n=6 for each group). (I) OG concentrations in serum and lung tissues after 14-day oral administration. Left: serum concentrations of OG across treatment groups. Right: lung tissue concentrations of OG, expressed as a percentage of tissue weight (n=3 for each group). * P<0.05, ** P<0.01, *** P<0.001. OG, glucoside; BLM, bleomycin; COL1A1, Collagen type I α 1 chain; α-SMA, α-smooth muscle actin; H&E, hematoxylin and eosin.

Article Snippet: Antibodies for Collagen type I α 1 chain (COL1A1; cat. no. 72026; 1:1,000), Phospho-STAT6 (Tyr641; cat. no. 56554S; 1:1,000) and α-smooth muscle actin (α-SMA; cat. no. 19245; 1:1,000) were obtained from Cell Signaling Technology, Inc.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture, Staining, Western Blot

Journal: Cell Insight

Article Title: Molecular basis of fox ACE2 recognition by receptor binding domains of SARS-CoV-2 and PCoV-GD

doi: 10.1016/j.cellin.2026.100314

Figure Lengend Snippet: Pseudovirus entry and cell membrane fusion mediated by SARS-CoV-2 and PCoV-GD spikes. (a) Western blotting detection of SARS-CoV-2 and PCoV-GD spike proteins in pseudovirus packaging cell lysate and pseudovirions. The SARS-CoV-2 and PCoV-GD spikes contain a C-terminal Strep-II tag and are detected using anti-Strep II antibody. For cell lysate, β-Actin is used as cellular internal control. For pseudovirions, HIV-1 p24 is used as viral internal control. (b) Western blotting detection of fACE2 and hACE2 protein in stable cell lines. The ACE2s contain a C-terminal Strep II tag and are detected using anti-Strep II antibody. β-Actin is used as cellular internal control. (c) The pseudovirus entry efficiency of SARS-CoV-2 and PCoV-GD into HEK293T, hACE2/HEK293T and fACE2/HEK293T cells. Pseudovirus without spike protein is used as negative control. Statistical comparisons are made using t -test. ∗∗∗, p < 0.001, ns: not significant. Fluorescence intensities are presented as mean ± SEM of three biological replicates. (d) Cell membrane fusion activity mediated by SARS-CoV-2 and PCoV-GD spikes assessed by DSP assay. Cells transfected with DSP 1-7 or DSP 8-11 are used as negative control. Fluorescence images are acquired after 8 h of co-incubation. Scale bars: 50 μm.

Article Snippet: After blocked with 5% (w/v) fat-free milk in tris-buffered saline (TBS) containing 0.05% (v/v) Tween 20 (TBST) at room temperature for 2 h, the membranes were incubated with primary antibodies against β-actin (Cat: LF201S, Epizyme Biotech), HIV-1 p24 protein (Cat: 11695-R002, SinoBiological) or the Strep II tag (Cat: AE066, Abclonal) in Protein-Free Blocking and Antibody Dilution Buffer (Beyotime Biotechnology) for 2 h. After washing in TBST buffer for three times, the membranes were then incubated with HRP-conjugated secondary antibodies (Abclonal) for 2h and later detected by a ChemiDoc MP Imaging System (Bio-Rad).

Techniques: Membrane, Western Blot, Control, Stable Transfection, Negative Control, Fluorescence, Activity Assay, Transfection, Incubation

Journal: Materials Today Bio

Article Title: Ultrasound-activated piezoelectric Silk-PVDF hydrogel reprograms the osteoimmune microenvironment via NRF2 signaling for accelerated bone regeneration

doi: 10.1016/j.mtbio.2026.102779

Figure Lengend Snippet: In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, Runx2, OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: Membranes were blocked with 5 % non-fat milk (BD DifcoTM, #232100) in TBST (Beyotime, #ST673) for 1 h at 25 °C and incubated overnight at 4 °C with the following primary antibodies: Osteopontin (OPN) : • Rabbit monoclonal (Proteintech, 22952-1-AP), 1:1000 • Collagen I (COL1A1): Rabbit polyclonal (Proteintech, 14695-1-AP), 1:1000 • RUNX2: Rabbit monoclonal (Proteintech, 20700-1-AP), 1:1000 • NRF2: Rabbit monoclonal (Proteintech, 16396-1-AP), 1:2000 • NQO1: Mouse monoclonal (Proteintech, 67240-1-Ig), 1:2000 • Beta Actin: Mouse monoclonal (Proteintech, 66009-1-Ig), 1:20,000 After three 10-min TBST washes, membranes were incubated with HRP-conjugated goat anti-rabbit/mouse IgG (Beyotime, #A0208/#A0216, 1:5000) for 2 h at 25 °C.

Techniques: In Vitro, Staining, Activity Assay, Quantitative RT-PCR, Expressing

Journal: Materials Today Bio

Article Title: Ultrasound-activated piezoelectric Silk-PVDF hydrogel reprograms the osteoimmune microenvironment via NRF2 signaling for accelerated bone regeneration

doi: 10.1016/j.mtbio.2026.102779

Figure Lengend Snippet: In vitro evaluation of osteogenesis related proteins and genes. (A) The relative protein expression levels of C1, Runx2 and OPN. (B–D) Semi-quantitative analysis of immunoblotting results of C1, Runx2 and OPN. (E) Immunofluorescence staining of OPN. (F) Immunofluorescence staining of OCN. (G) The relative protein expression levels of C1, Runx2 and OPN. (H–J) Semi-quantitative analysis of immunoblotting results of C1, Runx2 and OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: Membranes were blocked with 5 % non-fat milk (BD DifcoTM, #232100) in TBST (Beyotime, #ST673) for 1 h at 25 °C and incubated overnight at 4 °C with the following primary antibodies: Osteopontin (OPN) : • Rabbit monoclonal (Proteintech, 22952-1-AP), 1:1000 • Collagen I (COL1A1): Rabbit polyclonal (Proteintech, 14695-1-AP), 1:1000 • RUNX2: Rabbit monoclonal (Proteintech, 20700-1-AP), 1:1000 • NRF2: Rabbit monoclonal (Proteintech, 16396-1-AP), 1:2000 • NQO1: Mouse monoclonal (Proteintech, 67240-1-Ig), 1:2000 • Beta Actin: Mouse monoclonal (Proteintech, 66009-1-Ig), 1:20,000 After three 10-min TBST washes, membranes were incubated with HRP-conjugated goat anti-rabbit/mouse IgG (Beyotime, #A0208/#A0216, 1:5000) for 2 h at 25 °C.

Techniques: In Vitro, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Translational Neurodegeneration

Article Title: Bacteroides coprocola protects dopaminergic neurons in rotenone-induced Parkinson’s disease mouse model by modulating gut microbiota dysbiosis and inhibiting the NLRP3 signaling pathway

doi: 10.1186/s40035-026-00542-8

Figure Lengend Snippet: NaA and NaB treatment inhibits the NLRP3 signaling pathway in the LPS model. a Representative Western blotting bands of FFAR2, SIRT1, p-IκB-α, NF-κB, NLRP3 and caspase-1 in the siRNA-FFAR2 groups. b Representative Western blotting bands of FFAR3, SIRT1, p-IκB-α, NF-κB, NLRP3 andcaspase-1 in the siRNA-FFAR3 groups. c Representative Western blotting bands of FFAR2/3, SIRT1, p-IκB-α, NF-κB, NLRP3 and caspase-1 in the siRNA-FFAR2/3 groups. n = 5 for each group. LPS model = LPS + ATP

Article Snippet: The primary antibodies included anti-NLRP3 (1:1000, Adipogen, AG-20B-0014-C100, Liestal, Switzerland), anti-interleukin-6 (IL-6) (1:1000, Abclonal, A0286), anti-ZO-1 (1: 5000, Proteintech, 21773-1-AP, Wuhan, China), anti-Occludin (1:5000, Proteintech, 27260-1-AP), anti-NF-κB (1:1000, Abclonal, A19653), anti-sirtuin type 1 (SIRT1) (1:1000, CST, 9475, Danvers, MA), anti-caspase-1 (1:1000, CST, 24232), anti-ionized calcium-binding adapter molecule 1 (Iba-1) (1:1000, CST, 17198), anti-phospho-IκBα (1:5000, HUABIO, HA722770, Hangzhou, China), anti-free fatty acid receptor (FFAR) 2 (1:1000, Proteintech, 84544-1-RR), anti-FFAR3 (1:1000, HUABIO, ER63606), anti-Toll-like receptor 4 (TLR4) (1:1000, Abclonal, A5258), anti-myeloid differentiation primary response protein 88 (MyD88) (1:1000, CST, 4283), and anti-interleukin-1β (IL-1β) (1:1000, Thermo, P420B, Waltham, MA). β-Actin (1:100000, Abclonal, AC026) was used as the internal control.

Techniques: Western Blot