Journal: International Journal of Molecular Medicine

Article Title: BML-111 suppresses TGF-β1-induced lung fibroblast activation in vitro and decreases experimental pulmonary fibrosis in vivo

doi: 10.3892/ijmm.2018.3914

Figure Lengend Snippet: BML-111 decreased TGF-β1-induced NIH3T3 cell α-SMA expression in a dose-dependent manner. (A) NIH3T3 cells express rs1 and (B) FPR2. Cells were pretreated with a vehicle (0.035% ethanol) or BML-111 (1, 10, 100, 200 and 500 nM) for 30 min and then treated with TGF-β1 (5 ng/ml) for 24 h. (C) The expression of α-SMA was assessed using western blotting and (D) quantified. Similar results were obtained from at least 3 sections. Data are expressed as the mean ± standard deviation. # P<0.05 and ## P<0.01 vs. the vehicle group. * P<0.05 and ** P<0.01 vs. the TGF-β1 group in the absence of BML-111. Magnification, ×200. TGF-β1, Transforming growth factor-β1; α-SMA, smooth muscle α actin; rs1, related sequence 1; FPR2, formyl peptide receptor; marker 1, Trans DNA ladder (Tiangen Biotech, Co., Ltd., Beijing, China).

Article Snippet: Following permeabilization, washing and blocking, the cells were incubated with rabbit anti-FPR2 anti-bodies (1:100; cat. no. AFR-002; Alomone Labs, Jerusalem, Israel) at 4°C overnight and then washed and incubated with a fluorescent-labeled secondary antibody (fluorescein isothiocyanate-labelled goat anti-rabbit immunoglobulin G; 1:50; cat. no. AS1110; Aspen Biological, Wuhan, China) for 45 min at 37°C.

Techniques: Expressing, Western Blot, Standard Deviation, Sequencing, Marker

Journal: Journal of Translational Medicine

Article Title: Defective formyl peptide receptor 2/3 and annexin A1 expressions associated with M2a polarization of blood immune cells in patients with chronic obstructive pulmonary disease

doi: 10.1186/s12967-018-1435-5

Figure Lengend Snippet: Diminished FPR2/3 expressions of blood immune cells in COPD patients. The COPD patients at presentation had significantly lower intracellular FPR3 expressions of a M1 monocyte, b M2a monocyte, c NK cell, d NK T cell, e Th cell, and f Tc cell as compared with the healthy non-smokers. Cell surface FPR2 expressions on g Th and h Tc cells were also lower in the COPD patients. FPR3 expression of NK cell was positively correlated with i pre-bronchodilator FEV1/FVC ration and j FEV1 %predicted

Article Snippet: (A) Surface markers were measured by simultaneously staining for 30 min at 4 °C with directly conjugated monoclonal antibodies (mAb) as follows: PE Mouse Anti-Human CD56 (BD Pharmingen, USA), PE-CyTM5 Mouse Anti-Human CD3 (BD Pharmingen), CD4-PC7 (Beckman Coulter; USA), CD8-PC7, CD16-PC7, CD14-PC7 (Beckman Coulter), PerCP-Cy5.5 Mouse Anti-Human CD209 (BD Pharmingen), Anti-human CFS-conjugated FPR1 (R&D Systems; USA), PE-conjugated anti-human FPR2 (R&D Systems) or isotype control mAb. (B) For intracellular staining, cells were stained with anti-CD8a-PE-Cyanine5 and incubated with Cytofix/Cytoperm TM for 20 min at 4 °C, and washed with Perm/Wash Buffer.

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: Defective formyl peptide receptor 2/3 and annexin A1 expressions associated with M2a polarization of blood immune cells in patients with chronic obstructive pulmonary disease

doi: 10.1186/s12967-018-1435-5

Figure Lengend Snippet: Changes in FPR1/2/3 expressions of blood immune cells in COPD patients after 1-year treatment and with various clinical phenotypes. In 10 COPD patients after 1-year treatment, a FPR3 expression of M1 monocyte, b NK cell, and c Th cell, as well as d FPR2 expression on Th cell all showed significant elevation. e COPD patients with a high MMRC dyspnea scale had higher FPR1 expression on neutrophil, while f those with a history of frequent moderate exacerbation in the past 1 year had lower FPR2 expression on neutrophil. g FPR1 expression on neutrophil showed significant reduction after 1-year treatment. h Ex-smokers with COPD had higher FPR3 expression of NK T cell than current smokers with COPD after 1 year treatment. COPD patient receiving oral steroid treatment for more than 3 months had i higher FPR2 expression on Th cell and j higher FPR3 expression of M1 monocyte than those without oral steroid use. *p < 0.05 for comparisons between COPD patients with a specific phenotype and healthy non-smokers by ANOVA test. # p < 0.05 for comparison between COPD patients with and without s specific phenotype by ANOVA test. § p < 0.05 for comparisons between COPD patients with and without a specific management after 1-year follow-up

Article Snippet: (A) Surface markers were measured by simultaneously staining for 30 min at 4 °C with directly conjugated monoclonal antibodies (mAb) as follows: PE Mouse Anti-Human CD56 (BD Pharmingen, USA), PE-CyTM5 Mouse Anti-Human CD3 (BD Pharmingen), CD4-PC7 (Beckman Coulter; USA), CD8-PC7, CD16-PC7, CD14-PC7 (Beckman Coulter), PerCP-Cy5.5 Mouse Anti-Human CD209 (BD Pharmingen), Anti-human CFS-conjugated FPR1 (R&D Systems; USA), PE-conjugated anti-human FPR2 (R&D Systems) or isotype control mAb. (B) For intracellular staining, cells were stained with anti-CD8a-PE-Cyanine5 and incubated with Cytofix/Cytoperm TM for 20 min at 4 °C, and washed with Perm/Wash Buffer.

Techniques: Expressing, Comparison

Journal: Journal of Translational Medicine

Article Title: Defective formyl peptide receptor 2/3 and annexin A1 expressions associated with M2a polarization of blood immune cells in patients with chronic obstructive pulmonary disease

doi: 10.1186/s12967-018-1435-5

Figure Lengend Snippet: Differential serum levels of five FPR ligands, including serum amyloid A (SAA), cathelicidin (LL-37), annexin A1 (ANXA1), lipoxin A4 (LXA4), and resolving D1 (RvD1), in COPD patients. a The COPD patients at presentation had significantly lower serum ANXA1 levels, which remained at low levels after 1-year medical treatment. Serum ANXA1 levels were b positively correlated with FPR2 expression on T helper cells, c positively correlated with FPR3 expression of natural killer cells, d negatively correlated with the percentage of M1 monocyte, and e positively correlated with the percentage of M2a monocyte. f The COPD patients with a history of more than 1 moderate exacerbation in the past 1 year had significantly lower serum ANXA1 levels as compared with that in those without the history or the healthy subjects. g Serum LXA4 levels showed significant reduction and h RvD1 showed elevation after 1-year medical treatment. *p < 0.01 for comparison between COPD patients with a specific phenotype and healthy non-smokers by ANOVA test. # p < 0.05 for comparison between COPD patients with and without s specific phenotype by ANOVA test

Article Snippet: (A) Surface markers were measured by simultaneously staining for 30 min at 4 °C with directly conjugated monoclonal antibodies (mAb) as follows: PE Mouse Anti-Human CD56 (BD Pharmingen, USA), PE-CyTM5 Mouse Anti-Human CD3 (BD Pharmingen), CD4-PC7 (Beckman Coulter; USA), CD8-PC7, CD16-PC7, CD14-PC7 (Beckman Coulter), PerCP-Cy5.5 Mouse Anti-Human CD209 (BD Pharmingen), Anti-human CFS-conjugated FPR1 (R&D Systems; USA), PE-conjugated anti-human FPR2 (R&D Systems) or isotype control mAb. (B) For intracellular staining, cells were stained with anti-CD8a-PE-Cyanine5 and incubated with Cytofix/Cytoperm TM for 20 min at 4 °C, and washed with Perm/Wash Buffer.

Techniques: Expressing, Comparison