Journal: International Journal of Molecular Medicine

Article Title: Hypercapnia exacerbates the disruption of the blood‑brain barrier by inducing interleukin‑1β overproduction in the blood of hypoxemic adult rats

doi: 10.3892/ijmm.2020.4604

Figure Lengend Snippet: Hypercapnia increased the expression of IL-1β in the hypoxic blood in vivo and in vitro (n=4). (A) There was an interaction effect between hypoxia treatment and hypercapnia treatment (P<0.01). (B) Simple effects analyses revealed increased IL-1β expression in the hypoxemia group ( ** P<0.01), but not in the hypercapnia group (P>0.05) compared with the Sham group. The expression levels of IL-1β were the highest in the HH group in comparison with the hypoxemia group ( ** P<0.01) and the hypercapnia group ( ** P<0.01). (C) There was an interaction effect between 0.2% O 2 treatment and 10% CO 2 treatment. (D) Simple effects analyses revealed increased IL-1β expression in the hypoxia group ( ** P<0.01), but not in the HC group (P>0.05) compared with the control group. The expression levels of IL-1β were the highest in the hypoxia + HC group compared with the hypoxia group ( ** P<0.01) and the HC group ( ** P<0.01). The concentrations of O 2 and CO 2 in the air were 21 and 0.03%, respectively. IL-1β, interleukin-1β; HH, hypercapnia + hypoxemia; ns, non‑significant; HC, high concentration of carbon dioxide.

Article Snippet: The IL-1β + IL-1Ra group was treated with IL-1β (40 ng/ml) and IL-1Ra (40 ng/ml; MedChemExpress; cat. no. HY-P7029).

Techniques: Expressing, In Vivo, In Vitro, Comparison, Control, Concentration Assay

Journal: International Journal of Molecular Medicine

Article Title: Hypercapnia exacerbates the disruption of the blood‑brain barrier by inducing interleukin‑1β overproduction in the blood of hypoxemic adult rats

doi: 10.3892/ijmm.2020.4604

Figure Lengend Snippet: IL-1β treatment increased IL-1R1 and p-IRAK-1 expression in RBECs (n=4). (A) Immunoreactive bands of IL-1R1 (80 kDa) and β-actin (42 kDa). (B) The bar graph shows increased IL-1R1 expression in the IL-1β group compared with that in the control group ( ** P<0.01). (C) Immunofluorescence images showing the expression of CD31 + RBECs (a and d, green), IL-1R1 (b and e, red), and the co-localization of IL-1R1 and RBECs (c and f). Enhanced IL-1R1 immunofluorescence was evident in the IL‑1β group compared with the control group. Scale bars (a-f): 10 μ m. (D) Immunoreactive bands of p-RIAK-1 (77 kDa), t-IRAK-1 (77 kDa) and β-actin (42 kDa). (E) The bar graph shows increased p-IRAK-1 expression in the IL-1β group compared with the control group ( ** P<0.01). The protein expression of p‑IRAK‑1 was significantly suppressed with IL‑1Ra treatment ( * P<0.05). RBECs, rat brain capillary endothelial cells; IL-1β, interleukin-1β; IL‑1R, interleukin‑1 receptor; IL‑1Ra, interleukin‑1 receptor antagonist; HC, high concentration of carbon dioxide; ns, non‑significant.

Article Snippet: The IL-1β + IL-1Ra group was treated with IL-1β (40 ng/ml) and IL-1Ra (40 ng/ml; MedChemExpress; cat. no. HY-P7029).

Techniques: Expressing, Control, Immunofluorescence, Concentration Assay

Journal: International Journal of Molecular Medicine

Article Title: Hypercapnia exacerbates the disruption of the blood‑brain barrier by inducing interleukin‑1β overproduction in the blood of hypoxemic adult rats

doi: 10.3892/ijmm.2020.4604

Figure Lengend Snippet: IL-1β treatment decreased the expression of tight junctional proteins in RBECs (n=4). (A) Immunoreactive bands of ZO-1 (220 kDa), occludin (59 kDa), claudin-5 (23 kDa) and β-actin (42 kDa). (B) There was decreased tight junctional protein expression in the IL-1β group compared with the control group (ZO-1: ** P<0.01; occludin: ** P<0.01; and claudin-5: ** P<0.01). The expression of tight junctional proteins was significantly upregulated following IL-1Ra treatment (ZO-1: ** P<0.01; occludin: ** P<0.01; and claudin-5: * P<0.05). RBECs, rat brain capillary endothelial cells; ZO-1, zonula occludens-1; IL-1β, interleukin-1β; IL‑1Ra, interleukin‑1 receptor antagonist; ns, non‑significant.

Article Snippet: The IL-1β + IL-1Ra group was treated with IL-1β (40 ng/ml) and IL-1Ra (40 ng/ml; MedChemExpress; cat. no. HY-P7029).

Techniques: Expressing, Control

Journal: Inflammation and Regeneration

Article Title: Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury

doi: 10.1186/s41232-024-00319-4

Figure Lengend Snippet: Early-stage tissue evaluation after neurite bundle-derived artificial nerve transplantation. a , b Immunohistochemistry images with anti-NFH and anti-CD31 antibodies. Dotted arrows indicate the proximal side of the intact nerve areas, and solid arrows indicate the NFH- and CD31-positive regeneration tip in the reconstructed tissue. Scale bars = 1 mm. c , d Quantitative comparison of axonal extension distance was indicated by NFH-positive areas and that of vascular elongation distance was demonstrated by CD31-positive areas at 1 and 2 weeks after transplantation. Significant differences between groups elongation distances were observed in 2-week samples only. e , f Iba1 immunohistochemistry image of the proximal part of the regenerated tissue at 2 weeks after transplantation (e). Scale bars = 1 mm. Quantitative analysis of Iba1-positive areas demonstrated that a larger number of macrophages aggregated in the Auto, motor TP and sensory TP (f). ( n = 3). * p < 0.05, ** p < 0.01, N.S. = not significant. Data are represented as the mean ± SEM

Article Snippet: The primary antibodies used in this study were human polyclonal anti-pan-ELAVL (ELAV-like protein 2/3/4) antibody (1:2000, kindly provided from Prof. Robert Darnell, Rockefeller University), rabbit polyclonal anti-TrkA antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), chicken polyclonal anti-TrkB antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), goat polyclonal anti-TrkC antibody (AF373, 1:200, R&D systems), rabbit polyclonal anti-parvalbumin antibody (PV-28, 1:1000, Swant), rabbit polyclonal anti-CGRP antibody (BML-CA1134, 1:500, Enzo Life Sciences), Goat polyclonal anti-Choline acetyltransferase antibody (AB144P, 1:200, Chemicon), Mouse monoclonal anti-Islet1 and Islet2 antibody (39.4D5, 1:200, DSHB), rabbit polyclonal anti-Neurofilament heavy polypeptide antibody (ab8135, 1:500, Abcam), goat polyclonal anti-mouse and anti-rat CD31 (AF3628, 1:100, R&D), rabbit polyclonal anti-Iba1 antibody (GtX100042, 1:500, GeneTex), goat anti-GFP antibody (600–101-215, 1:1000, Rockland).

Techniques: Derivative Assay, Transplantation Assay, Immunohistochemistry, Comparison

Journal: Journal of Inflammation Research

Article Title: Glycochenodeoxycholic acid induces the release of IL-1beta from L02 cells via the NLRP3/caspase-1/GSDMD pathway to activate LX2 cells

doi: 10.2147/JIR.S532042

Figure Lengend Snippet: GCDCA-mediated hepatocyte IL-1β secretion activates HSCs. ( A ) Western blot analysis ofα-SMA and Collagen-I in LX2 cells treated with conditioned medium (CM) from GCDCA/LPS-stimulated L02 hepatocytes. ( B ) The protein levels of α-SMA and Collagen-I in LX2 cells with treatment of IL-1 receptor antagonist (IL-1RA, 200 ng/mL) upon CM exposure. ( C and D ) Representative images and quantification of LX2 migration upon CM exposure detected by wound healing assay. ( E ) CCK-8 assay of LX2 proliferation by CM treatment. n ≥ 3 for each group. a P < 0.05.

Article Snippet: Conditioned media from treated L02 cells were co-cultured with LX2 cells in the presence/absence of Interleukin-1 receptor antagonist (IL-1RA) (200 ng/mL, HY-P7029A, MCE).

Techniques: Western Blot, Migration, Wound Healing Assay, CCK-8 Assay

Journal: Small Methods

Article Title: Mannose Oligosaccharide‐Conjugated In Situ Pore‐Forming Injectable Hydrogels for Rheumatoid Arthritis Treatment by Reprogramming Macrophage Extracellular Vesicles

doi: 10.1002/smtd.202500605

Figure Lengend Snippet: H/CS‐MOS hydrogels regulate the recruitment and polarization of macrophages. a–c) Flow cytometric analysis showing the proportion of the macrophages among the recruited cells (a) and the proportion of the M2 macrophages among all the macrophages (b) along with their quantitative comparison (c) ( n = 3). d–f) Flow cytometry analysis of the polarization of the macrophages, indicated by CD197 (M1 phenotype, d) and CD206 (M2 phenotype, e), along with their quantitative comparison (f) ( n = 3). g) Flow cytometry analysis showing the polarization shift of M1 macrophages (induced with LPS and IFN‐γ) cultured on the different scaffolds, along with their quantitative comparison ( n = 3). h,i) Western blot analysis of protein levels of CD206, CD44, pro‐inflammatory factors (IFN‐γ and TNF‐α), anti‐inflammatory factor (IL‐1Ra), and Arg‐1, with quantification normalized to the HA group ( n = 3). j) Quantitative analysis of inflammatory cytokine levels of IL‐6, IL‐1β, TNF‐α, IL‐10 and TGF‐β ( n = 3). All data are presented as mean ± SD. (The p values were determined using unpaired two‐tailed t tests for two‐group comparisons and one‐way ANOVA followed by Tukey's post hoc test for multiple‐group comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, # p < 0.05, ## p < 0.01, ### p < 0.001, # indicates a comparison with the HA group within the same protein experimental group.

Article Snippet: The membrane pieces were probed overnight at 4 °C with primary antibodies against GAPDH (#2118, Cell Signaling Technology, CST, Danvers, MA, USA), CD206 (ab64693, Abcam), CD44 (ab157107, Abcam), IFN‐γ (ab198801, Abcam), TNF‐α (ab307164, Abcam), IL‐1Ra (NBP1‐32568, Novus Biologicals, Littleton, CO, USA) and Arg‐1 (#93668, CST).

Techniques: Comparison, Flow Cytometry, Cell Culture, Western Blot, Two Tailed Test