Journal: Oncology Letters

Article Title: Role of T helper 17 cytokines in the tumour immune inflammation response of patients with laryngeal squamous cell carcinoma

doi: 10.3892/ol.2017.6253

Figure Lengend Snippet: Positive and negative expression of IL17/IL17R immunohistochemistry in tissue microarray sections. (A) Positive and (B) negative expression of IL17. (C) Positive and (D) negative expression of IL17R. Original magnification, ×20. IL, interleukin; R, receptor.

Article Snippet: The following anti-human monoclonal antibodies were used: Rabbit anti-IL17 (#bs-2140R; BIOSS, Beijing, China) and rabbit anti-IL17A receptor (IL17R; #bs-2606R; BIOSS).

Techniques: Expressing, Immunohistochemistry, Microarray

Journal: Oncology Letters

Article Title: Role of T helper 17 cytokines in the tumour immune inflammation response of patients with laryngeal squamous cell carcinoma

doi: 10.3892/ol.2017.6253

Figure Lengend Snippet: Positive results of IL17/IL17R immunohistochemistry in tumors and controls.

Article Snippet: The following anti-human monoclonal antibodies were used: Rabbit anti-IL17 (#bs-2140R; BIOSS, Beijing, China) and rabbit anti-IL17A receptor (IL17R; #bs-2606R; BIOSS).

Techniques: Immunohistochemistry

Journal: Oncology Letters

Article Title: Role of T helper 17 cytokines in the tumour immune inflammation response of patients with laryngeal squamous cell carcinoma

doi: 10.3892/ol.2017.6253

Figure Lengend Snippet: Reverse transcription-quantitative polymerase chain reaction analysis of the Th17-associated intracellular cytokines and transcription factors IL17, IL23 and RORγt in tumors and controls. *P<0.05. IL, interleukin; ROR, RAR-related orphan receptor; Cq, quantification cycle.

Article Snippet: The following anti-human monoclonal antibodies were used: Rabbit anti-IL17 (#bs-2140R; BIOSS, Beijing, China) and rabbit anti-IL17A receptor (IL17R; #bs-2606R; BIOSS).

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of Neuroinflammation

Article Title: RhANP attenuates endotoxin-derived cognitive dysfunction through subdiaphragmatic vagus nerve-mediated gut microbiota–brain axis

doi: 10.1186/s12974-021-02356-z

Figure Lengend Snippet: Roles of SDV in rhANP-mediated reduction of LPS-induced systemic inflammation and neuroinflammation. a Treatment schedule. Mice were intraperitoneally injected with lipopolysaccharides (LPS, 5 mg/kg). Recombinant human ANP (rhANP; 1.0 mg/kg) or 0.9% saline were intraperitoneally injected to mice at 24 h before and 10 min after LPS injection. Subdiaphragmatic vagotomy (SDV) was performed 14 days prior to LPS injection. Plasma and hippocampus were collected 24 h after LPS injection. b Body weight loss in each group (two-way ANOVA: rhANP: F 1,36 = 7.058, P = 0.0117; SDV: F 1,36 = 12.72, P = 0.0010; interaction: F 1,36 = 4.923, P = 0.0329). The plasma levels of interleukin (IL)-6 ( c ; two-way ANOVA: rhANP: F 1,36 = 13.07, P = 0.0009; SDV: F 1,36 = 15.60, P = 0.0003; interaction: F 1,36 = 20.51, P < 0.0001), IL-17A ( d ; two-way ANOVA: rhANP: F 1,36 = 6.111, P = 0.0183; SDV: F 1,36 = 7.794, P = 0.0083; interaction: F 1,36 = 6.027, P = 0.0191), interferon (IFN)-γ ( e ; two-way ANOVA: rhANP: F 1,36 = 6.460, P = 0.0155; SDV: F 1,36 = 8.447, P = 0.0062; interaction: F 1,36 = 7.836, P = 0.0082), and tumor necrosis factor (TNF)-α ( f ; two-way ANOVA: rhANP: F 1,36 = 4.828, P = 0.0345; SDV: F 1,36 = 6.217, P = 0.0174; interaction: F 1,36 = 7.883, P = 0.0080). Western blot analysis of ionized calcium-binding adapter molecule 1 (iba-1) ( g ; two-way ANOVA: rhANP: F 1,36 = 6.208, P = 0.0175; SDV: F 1,36 = 4.772, P = 0.0355; interaction: F 1,36 = 4.996, P = 0.0317), IL-6 ( h ; two-way ANOVA: rhANP: F 1,36 = 6.286, P = 0.0168; SDV: F 1,36 = 6.894, P = 0.0126; interaction: F 1,36 = 8.569, P = 0.0059), IL-17A ( i ; two-way ANOVA: rhANP: F 1,36 = 4.268, P = 0.0461; SDV: F 1,36 = 3.363, P = 0.0750; interaction: F 1,36 = 5.503, P = 0.0246), interferon (IFN)-γ ( j ; two-way ANOVA: rhANP: F 1,36 = 4.706, P = 0.0367; SDV: F 1,36 = 5.685, P = 0.0225; interaction: F 1,36 = 3.530, P = 0.0684), TNF-α ( k ; two-way ANOVA: rhANP: F 1,36 = 1.598, P = 0.2144; SDV: F 1,36 = 12.72, P = 0.0010; interaction: F 1,36 = 4.345, P = 0.0443), inducible nitric oxide synthase (iNOS) ( l ; two-way ANOVA: rhANP: F 1,36 = 4.764, P = 0.0357; SDV: F 1,36 = 2.933, P = 0.0954; interaction: F 1,36 = 5.462, P = 0.0251) and their respective β-actin in the hippocampus. Data are shown as mean ± SEM, n = 10/group. * P < 0.05, ** P < 0.01, *** P < 0.0001; N.S. not significant

Article Snippet: The membranes were blocked in 5% non-fat dried milk for 1 h at room temperature, and then incubated with the following primary antibodies: rabbit polyclonal anti-ionized calcium-binding adapter molecule 1 (iba-1; 1:1000, #016-20001: Wako Pure Chemical Industries, Ltd., Tokyo, Japan), rabbit monoclonal anti-IL-6 (1:1000, #12912S, Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit monoclonal anti-IL-17A (1:1000, #A0688, ABclonal, Wuhan, China), rabbit monoclonal anti-IFN-γ (1:1000, #A12450, ABclonal, Wuhan, China), rabbit polyclonal anti-TNF-α (1:1000, #A0277, ABclonal, Wuhan, China), rabbit monoclonal anti-inducible nitric oxide synthase (iNOS; 1:1000, #ab3523, Abcam, Cambridge, MA, USA), brain-derived neurotrophic factor (BDNF; 1:1000, #A4873, ABclonal, Wuhan, China), rabbit polyclonal phosphorylated tyrosine kinase receptor B (p-TrkB; 1:1000, #AP0423, ABclonal, Wuhan, China), rabbit polyclonal total TrkB (t-TrkB; 1:1000, #A2099, ABclonal, Wuhan, China), and mouse monoclonal anti-β-actin (1:1000, #A2319, ABclonal, Wuhan, China) overnight at 4 ℃.

Techniques: Injection, Recombinant, Western Blot, Binding Assay

Journal: Journal of Neuroinflammation

Article Title: RhANP attenuates endotoxin-derived cognitive dysfunction through subdiaphragmatic vagus nerve-mediated gut microbiota–brain axis

doi: 10.1186/s12974-021-02356-z

Figure Lengend Snippet: Effects of rhANP on plasma inflammatory cytokines after LPS-triggered endotoxemia. a Treatment schedule. Mice were intraperitoneally injected with lipopolysaccharides (LPS, 5 mg/kg) or 0.9% saline. Recombinant human ANP (rhANP; 1.0 mg/kg) or 0.9% saline were intraperitoneally injected to mice 24 h before and 10 min after LPS injection. Spleen and plasma were collected 24 h after injection of LPS or 0.9% saline. b Body weight loss in mice treated with rhANP or 0.9% saline 24 h after injection of LPS or 0.9% saline (one-way ANOVA: F 2,27 = 58.21, P < 0.0001). c Representative picture of spleen and spleen weight (one-way ANOVA: F 2,28 = 27.53, P < 0.0001). d Ratio of spleen weight/body weight (one-way ANOVA: F 2,28 = 20.17, P < 0.0001). e Plasma levels of interleukin (IL)-6 (one-way ANOVA: F 2,28 = 65.35, P < 0.0001). f Plasma levels of IL-17A (one-way ANOVA: F 2,28 = 16.82, P < 0.0001). g Plasma levels of interferon (IFN)-γ (one-way ANOVA: F 2,28 = 12.62, P < 0.0001). h Plasma levels of tumor necrosis factor (TNF)-α (one-way ANOVA: F 2,28 = 51.43, P < 0.0001). i There was a positive correlation (r = 0.797, P < 0.001) between spleen weight and plasma IL-6. j There was a positive correlation ( r = 0.629, P < 0.001) between spleen weight and plasma IL-17A. k There was a positive correlation ( r = 0.479, P = 0.006) between spleen weight and plasma IFN-γ. l Positive correlation ( r = 0.637, P < 0.001) between spleen weight and plasma TNF-α was observed. Data are shown as mean ± SEM, n = 10 or 11/group. ** P < 0.01, *** P < 0.0001

Article Snippet: The membranes were blocked in 5% non-fat dried milk for 1 h at room temperature, and then incubated with the following primary antibodies: rabbit polyclonal anti-ionized calcium-binding adapter molecule 1 (iba-1; 1:1000, #016-20001: Wako Pure Chemical Industries, Ltd., Tokyo, Japan), rabbit monoclonal anti-IL-6 (1:1000, #12912S, Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit monoclonal anti-IL-17A (1:1000, #A0688, ABclonal, Wuhan, China), rabbit monoclonal anti-IFN-γ (1:1000, #A12450, ABclonal, Wuhan, China), rabbit polyclonal anti-TNF-α (1:1000, #A0277, ABclonal, Wuhan, China), rabbit monoclonal anti-inducible nitric oxide synthase (iNOS; 1:1000, #ab3523, Abcam, Cambridge, MA, USA), brain-derived neurotrophic factor (BDNF; 1:1000, #A4873, ABclonal, Wuhan, China), rabbit polyclonal phosphorylated tyrosine kinase receptor B (p-TrkB; 1:1000, #AP0423, ABclonal, Wuhan, China), rabbit polyclonal total TrkB (t-TrkB; 1:1000, #A2099, ABclonal, Wuhan, China), and mouse monoclonal anti-β-actin (1:1000, #A2319, ABclonal, Wuhan, China) overnight at 4 ℃.

Techniques: Injection, Recombinant

Journal: Journal of Neuroinflammation

Article Title: RhANP attenuates endotoxin-derived cognitive dysfunction through subdiaphragmatic vagus nerve-mediated gut microbiota–brain axis

doi: 10.1186/s12974-021-02356-z

Figure Lengend Snippet: Effects of rhANP on the neuroinflammation and cognitive function after LPS-triggered endotoxemia. a Treatment schedule. Mice were intraperitoneally injected with lipopolysaccharides (LPS, 5 mg/kg) or 0.9% saline. Recombinant human ANP (rhANP; 1.0 mg/kg) or 0.9% saline were intraperitoneally injected to mice 24 h before and 10 min after LPS injection. The prefrontal cortex (PFC) and hippocampus were collected 24 h after injection of LPS or 0.9% saline. b , c Western blot analysis of ionized calcium-binding adapter molecule 1 (iba-1) in the prefrontal cortex (PFC) (one-way ANOVA: F 2,27 = 6.170, P = 0.0062) and hippocampus (one-way ANOVA: F 2,27 = 5.250, P = 0.0119). d , e Western blot analysis of interleukin (IL)-6 in the PFC (one-way ANOVA: F 2,27 = 5.958, P = 0.0072) and hippocampus (one-way ANOVA: F 2,27 = 17.60, P < 0.0001). f , g Western blot analysis of IL-17A in the PFC (one-way ANOVA: F 2,27 = 4.498, P = 0.0206) and hippocampus (one-way ANOVA: F 2,27 = 6.268, P = 0.0058). h , i Western blot analysis of interferon (IFN)-γ in the PFC (one-way ANOVA: F 2,27 = 11.18, P = 0.0003) and hippocampus (one-way ANOVA: F 2,27 = 7.611, P = 0.0024). j , k Western blot analysis of tumor necrosis factor (TNF)-α in the PFC (one-way ANOVA: F 2,27 = 5.530, P = 0.0097) and hippocampus (one-way ANOVA: F 2,27 = 8.550, P = 0.0013). l , m Western blot analysis of inducible nitric oxide synthase (iNOS) in the PFC (one-way ANOVA: F 2,27 = 7.328, P = 0.0029) and hippocampus (one-way ANOVA: F 2,27 = 7.597, P = 0.0024). n , o Entries in the novel arm (one-way ANOVA: F 2,27 = 31.33, P < 0.0001) and duration in the novel arm (one-way ANOVA: F 2,27 = 13.34, P < 0.0001) in the Y maze test. p Latency to eat food in the buried food test (one-way ANOVA: F 2,27 = 7.129, P = 0.0033). Data are shown as mean ± SEM, n = 10/group. * P < 0.05, ** P < 0.01, *** P < 0.0001; N.S. not significant

Article Snippet: The membranes were blocked in 5% non-fat dried milk for 1 h at room temperature, and then incubated with the following primary antibodies: rabbit polyclonal anti-ionized calcium-binding adapter molecule 1 (iba-1; 1:1000, #016-20001: Wako Pure Chemical Industries, Ltd., Tokyo, Japan), rabbit monoclonal anti-IL-6 (1:1000, #12912S, Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit monoclonal anti-IL-17A (1:1000, #A0688, ABclonal, Wuhan, China), rabbit monoclonal anti-IFN-γ (1:1000, #A12450, ABclonal, Wuhan, China), rabbit polyclonal anti-TNF-α (1:1000, #A0277, ABclonal, Wuhan, China), rabbit monoclonal anti-inducible nitric oxide synthase (iNOS; 1:1000, #ab3523, Abcam, Cambridge, MA, USA), brain-derived neurotrophic factor (BDNF; 1:1000, #A4873, ABclonal, Wuhan, China), rabbit polyclonal phosphorylated tyrosine kinase receptor B (p-TrkB; 1:1000, #AP0423, ABclonal, Wuhan, China), rabbit polyclonal total TrkB (t-TrkB; 1:1000, #A2099, ABclonal, Wuhan, China), and mouse monoclonal anti-β-actin (1:1000, #A2319, ABclonal, Wuhan, China) overnight at 4 ℃.

Techniques: Injection, Recombinant, Western Blot, Binding Assay

Journal: Neural Regeneration Research

Article Title: Fingolimod protects against neurovascular unit injury in a rat model of focal cerebral ischemia/reperfusion injury

doi: 10.4103/1673-5374.353500

Figure Lengend Snippet: Fingolimod (FTY-720) attenuates ischemia/reperfusion-induced glial cell activation and IL-17A expression. (A) Representative immunofluorescence staining for IL-17A (red) and GFAP (green). (B) Representative immunofluorescence staining for IL-17A (red) and Iba-1 (green). Scale bars: 50 μm. (C) Representative western blot bands for IL-17A. (D) Quantification of IL-17A protein expression ( n = 5 rats per group, * P < 0.05, vs . DMSO group, one-way analysis of variance followed by Bonferroni post hoc test). The experiment was repeated five times. DMSO: Dimethyl sulfoxide; GFAP: glial fibrillary acidic protein; Iba1: ionized calcium-binding adapter molecule 1; IL: interleukin.

Article Snippet: The membranes were blocked for 1 hour with 5% skimmed milk (Solarbio Science & Technology Co., Ltd.) in TBST (tris-buffered saline and Polysorbate 20, also known as Tween 20) at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: rabbit anti-occludin antibody (tight junction protein, 1:1000; Cat# 40-4700, Thermo Fisher Scientific), rabbit anti-claudin-5 polyclonal antibody (tight junction protein, 1:1000; Cat# PA5-99415, Thermo Fisher Scientific), rabbit anti-S1PR1 polyclonal antibody (endothelial cell receptor; 1:1000; Cat# ab11424, Abcam), rabbit anti-IL-17A polyclonal antibody (an inflammatory cytokine; 1:1000; Cat# PA5-106856, Thermo Fisher Scientific), mouse anti-β-actin polyclonal antibody (1:3000; Cat# 3700, CST).

Techniques: Activation Assay, Expressing, Immunofluorescence, Staining, Western Blot, Binding Assay