Journal: ImmunoTargets and Therapy

Article Title: Recombinant Antithrombin Alleviated Pulmonary Injury and Inflammation in LPS-Induced ARDS by Inhibiting IL17a/NF-κB Signaling

doi: 10.2147/ITT.S502925

Figure Lengend Snippet: rAT reduced pulmonary inflammation in LPS-induced ARDS model mice. ( A–D ) The levels of inflammatory factors, including IL-6 ( A ), TNF-α ( B ), IL-8 ( C ), and hs-CRP ( D ), were decreased in the serum of the rAT-treated group, as detected by ELISA. ( E ) Immunohistochemical staining for F4/80 revealed that rAT treatment reduced the proportion of macrophages in the lung tissue of LPS-induced ARDS model mice. ( F ) Immunohistochemical staining for MRCI, a marker of M2 macrophages, showed that rAT treatment increased the proportion of M2 macrophages in the lung tissue of LPS-induced ARDS model mice. ( G ) Immunohistochemical staining for Ly6G, a marker of neutrophils, showed that rAT treatment reduced the proportion of neutrophils in the lung tissue of LPS-induced ARDS model mice. All the data are presented as the means ± SDs of three independent experiments. One-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, ns, not significant; scale bar, 50 μm.

Article Snippet: A mouse CXCL15 ELISA Kit (E-EL-M0269) and a mouse hs-CRP ELISA Kit (E-EL-M0677) were purchased from Elabscience (Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Marker

Journal: ImmunoTargets and Therapy

Article Title: Recombinant Antithrombin Alleviated Pulmonary Injury and Inflammation in LPS-Induced ARDS by Inhibiting IL17a/NF-κB Signaling

doi: 10.2147/ITT.S502925

Figure Lengend Snippet: The efficacy of rAT in mitigating lung injury, suppressing the immune response, and inhibiting the activation of the NF-κB signaling pathway in LPS-induced ARDS mice were diminished by the administration of IL-17a. ( A ) ELISA results demonstrated that the administration of IL17a inhibited the ability of rAT to reduce inflammatory factors, including IL-6, TNF-α, and IL-8, in the serum of LPS-induced ARDS mice. ( B ) The analysis of the wet/dry weight ratio of the lung tissue revealed that the administration of IL17a counteracted the ability of rAT to alleviate pulmonary exudation in LPS-induced ARDS mice. ( C ) The administration of IL17a did not significantly affect the ability of rAT to reduce the number of cells in the BALF of LPS-induced ARDS mice. ( D ) The administration of IL17a attenuated the ability of rAT to reduce the concentrations of proteins in the BALF of LPS-induced ARDS mice. ( E ) Real-time PCR results showed that the administration of IL17a blocked the ability of rAT to downregulate the expression of target genes in the IL17a/NF-κB signaling pathway. ( F ) The protein levels of the NF-κB signaling pathway were assessed by Western blotting, and gray intensity analysis of the blots showed that the administration of IL17a in LPS-induced ARDS mice counteracted the ability of rAT to suppress the phosphorylation of IκBα, IKKα/β, and P65. The data are expressed as the means ± SDs (n=3 in each group). One-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, and ns not significant.

Article Snippet: A mouse CXCL15 ELISA Kit (E-EL-M0269) and a mouse hs-CRP ELISA Kit (E-EL-M0677) were purchased from Elabscience (Wuhan, China).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Phospho-proteomics

Journal: International journal of molecular sciences

Article Title: Surgical Site-Released Tissue Is Potent to Generate Bone onto TCP and PCL-TCP Scaffolds In Vitro.

doi: 10.3390/ijms242115877

Figure Lengend Snippet: Figure 6. (a–c) ELISA boxplots of the osteogenic markers osteocalcin (a), osteopontin (b), and collagen I (c). Boxplots compare the concentration of ocn, opn, and col I in the control group with the PCL-TCP and β-TCP group over the cultivation time of 6 weeks. Groups are further subdivided into the condition’s agarose and glue, to examine if the chosen adhesive, used for the attachment of the scaffolds onto the plastic wells, makes a difference in protein expression. The boxblots indicating the median within the 25–75% percentile. The dots represent single outliners during measurements.

Article Snippet: Enzyme-linked immunosorbent assays (ELISAs) (human Duoset ELISA kit, R&D Systems, Bio Techne, Minneapolis, MN, USA) were then performed using the supernatants at a dilution of 1:10 for osteocalcin (1:50; pure for collagen and pure for osteopontin) according to the manufacturer’s protocol and measured at 450 nm using a Multiscan Ascent reader (Thermo Scientific).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Control, Adhesive, Expressing

Journal: Bioengineering & Translational Medicine

Article Title: Neuron‐targeted 2‐deoxyglucose‐dendrimer‐rosiglitazone nanotherapy mitigates neuroinflammation and cognitive deficits in pediatric traumatic brain injury

doi: 10.1002/btm2.70053

Figure Lengend Snippet: FIGURE 8 mRNA expressions of pro- and anti-inflammatory markers and cell death markers from sham (n = 12, 6 M/6 F), traumatic brain injury (TBI) + vehicle (n = 12, 6 M/6 F), TBI + rosiglitazone (Rosi) (n = 12, 6 M/6 F), and TBI + 2-deoxyglucose (2DG-D)-Rosi (n = 12, 6 M/6 F) groups. Neurons were isolated from the injured brain regions (or the matching area from the sham mice) at 24-h post-treatment for gene expression evaluations. Data were presented as mean ± SEM. Treatment groups were presented as sham (gray circles), TBI + vehicle (blue circles), TBI + Rosi (magenta circles), and TBI + 2DG-D-Rosi (green circles). (a–d) The expression of pro-inflammatory makers, tumor necrosis factor-alpha (TNF-α) (a), interleukin-6 beta (IL-1β) (b), Toll-like receptor 4 (TLR4) (c), and NLR Family Pyrin Domain Containing 3 (NLRP3) (d). (e) The expression of anti-inflammatory maker IL-13. (f) The expression of cell death marker Fas. *p < 0.05; **p < 0.01; ***p < 0.001. (g) The co- localization of neurodegenerative markers marker Fluoro-Jade C (FJC) (green) and neuronal marker (NeuN, red) was evaluated in sham (n = 8, 4 M/4 F), TBI + vehicle (n = 8, 4 M/4 F), TBI + Rosi (n = 8, 4 M/4 F), and TBI + 2DG-D-Rosi (n = 8, 4 M/4 F) groups at 24-h post-injury. Images (40, 5 images/animal) were randomly acquired from the cortex area (mainly primary motor cortex and primary somatosensory cortex) in the injured brain regions (approximately between bregma +1 mm and bregma 0.5 mm) using the Nikon Eclipse TS2R fluorescent microscope (Nikon, NY, USA). The representative images from male treatment groups (upper panels) and female treatment groups (lower panels). 40,6-Diamidino-2-phenylindole (DAPI) (blue) was used for nucleus stains. Scale bars: 100 μm. (h) The quantification of FJC+NeuN+ cells among treatment groups. Data were presented as mean ± SEM. Treatment groups were presented as sham (gray circles), TBI + vehicle (blue circles), TBI + Rosi (magenta circles), and TBI + 2DG-D-Rosi (green circles). *p < 0.05; **p < 0.01; ****p < 0.0001.

Article Snippet: Brain sections were washed 3 times in PBS for 5 min each, incubated in 0.001% Fluoro-Jade C (FJC) (Cat# TR-160-FJC, Biosensis, CA, USA) for 10 min, washed six times in PBS for 15 min each, and incubated with rabbit anti-NeuN (1:250; Cat# ab177487; Abcam, MA, USA).

Techniques: Isolation, Gene Expression, Expressing, Marker, Microscopy