Journal: Immunity, inflammation and disease

Article Title: Dexamethasone and potassium canrenoate alleviate hyperalgesia by competitively regulating IL-6/JAK2/STAT3 signaling pathway during inflammatory pain in vivo and in vitro.

doi: 10.1002/iid3.721

Figure Lengend Snippet: FIGURE 1 Schematic of the experimental protocol. Nongenomic effect (rapid): usually occur within a few minutes via membrane‐ bound receptors. Genomic effect (delayed and long‐lasting): usually take several hours or days to manifest specific gene transcription. Cane, potassium canrenoate; Dexa, dexamethasone; ED50, median effective dose; ELISA, enzyme‐linked immunosorbent assay; FCA, Freund's complete adjuvant; IL‐6, interleukin 6; i.pl., intraplantar.

Article Snippet: Competing IL‐ 6 (0–2000 pg/ml) was mixed with 1mM Dexa (1 mg/ 2.548ml) and/or 1mM Cane (1mg/2.522 ml) and incubated at 37°C for 1 h. Then, the concentration of IL‐6 in the presence of 1 mM Dexa and/or Cane was determined using rat IL‐6 ELISA kit (Elabscience), according to the operating instruction.

Techniques: Membrane, Enzyme-linked Immunosorbent Assay, Adjuvant

Journal: Immunity, inflammation and disease

Article Title: Dexamethasone and potassium canrenoate alleviate hyperalgesia by competitively regulating IL-6/JAK2/STAT3 signaling pathway during inflammatory pain in vivo and in vitro.

doi: 10.1002/iid3.721

Figure Lengend Snippet: FIGURE 6 Competitive binding assay and the schematic summarizing the key findings of this study. (A) Indicated concentrations of competing IL‐6 (0–2000 pg/ml) were incubated with 1 mM Dexa and/or Cane at 37°C for 1 h. After incubation, the competitive binding of Dexa and/or Cane to IL‐6 was evaluated by competitive ELISA. The current data showed that Dexa had a higher affinity to IL‐6 than Cane at the same concentration. Data are expressed as mean ± SD (n = 6). *p < .05 versus IL‐6 + 1 mM Dexa + 1 mM Cane group; ▲p < .05 versus IL‐ 6 + 1 mM Cane group. Statistical comparisons were conducted with one‐way ANOVA followed by Bonferroni's test. (B) Dexa and Cane competitively engage in IL‐6‐mediated JAK2/STAT3 signaling pathway, in which Dexa was dominant in binding to IL‐6, thus regulating tactile allodynia during inflammatory pain development. Cane, potassium canrenoate; Dexa, dexamethasone; ELISA, enzyme‐linked immunosorbent assays; IL‐6, interleukin‐6; IL‐6R, interleukin‐6 receptor; JAK2, Janus kinase 2; STAT3, Signal transducer and activator of transcription 3.

Article Snippet: Competing IL‐ 6 (0–2000 pg/ml) was mixed with 1mM Dexa (1 mg/ 2.548ml) and/or 1mM Cane (1mg/2.522 ml) and incubated at 37°C for 1 h. Then, the concentration of IL‐6 in the presence of 1 mM Dexa and/or Cane was determined using rat IL‐6 ELISA kit (Elabscience), according to the operating instruction.

Techniques: Competitive Binding Assay, Incubation, Binding Assay, Competitive ELISA, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Theranostics

Article Title: Cancer-associated Fibroblast-derived IL-6 Promotes Head and Neck Cancer Progression via the Osteopontin-NF-kappa B Signaling Pathway

doi: 10.7150/thno.22182

Figure Lengend Snippet: Fibroblasts contribute to the up-regulation of OPN in HNC cells. (A) Immunohistochemical analysis of OPN protein expression in HNC tissues and matched adjacent normal tissues (Scale bar: 25 μm). Higher staining of OPN was observed in tumor cells of the edge of bulk tumors. (B) The mRNA and protein levels of OPN were determined in NFs, CAFs and cancer cells (derived from 4 HNC patients) using real-time PCR and western blotting. (C) OPN at the protein and mRNA level and in the supernatant was detected in 8 representative HNC cell lines and normal oral epithelial cells (titled normal) using western blotting, real-time PCR and ELISA. (D) The protein level, supernatant concentration and mRNA level of OPN were determined in CAL-27 and SCC-25 cells after co-culture with NFs. (E) The protein level, supernatant concentration and mRNA level of OPN were determined in NFs after co-culture with CAL-27 and SCC-25 cells. (* p <0.05; ** p <0.01; *** p <0.001; **** p <0.0001)

Article Snippet: The supernatant OPN and IL-6 concentrations and the plasma OPN and IL-6 levels in patients with HNC were assessed using the Human OPN ELISA kit (Boster, China) and human IL-6 ELISA kit (Boster, China).

Techniques: Immunohistochemical staining, Expressing, Staining, Derivative Assay, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay, Co-Culture Assay

Journal: Theranostics

Article Title: Cancer-associated Fibroblast-derived IL-6 Promotes Head and Neck Cancer Progression via the Osteopontin-NF-kappa B Signaling Pathway

doi: 10.7150/thno.22182

Figure Lengend Snippet: Effects of stromal IL-6-induced OPN on promoting tumor growth and metastasis in vivo . (A) OPN overexpression in CAL-27 cells facilitated the xenograft tumor growth in nude mice. (B) Plasma OPN levels of the tumor-bearing mice from the normal group, CAL-27 group and CAL-27-OPN group. (C) Subcutaneous injection of OPN antibody (10 μg per tumor nodule) or IL-6 antibody (10 μg per tumor nodule) around the tumor partly inhibited NF-mediated tumor growth, and the combination of OPN antibody (5 μg per tumor nodule) and IL-6 antibody (5 μg per tumor nodule) exhibited a more powerful antitumor activity. (D) Plasma OPN levels of the tumor-bearing mice from the normal group, IgG treatment group, OPN antibody treatment group, IL-6 antibody treatment group and the combination group. (E) Overexpression of OPN in Rca-T cells promoted the formation and growth of metastatic nodules in nude mice (Scale bar, left: 5 mm; right: 100 μm). (F) Western blot analysis confirmed the exogenous expression of OPN in Rca-T cells, and plasma OPN levels of the mice involved in experimental metastasis were measured using ELISA. (G) Kaplan-Meier analyses of overall survival. (H, I and J) ROC curve analysis of the mRNA panel of OPN and IL-6 stratified by different groups in the validation set. ROC plots for the mRNA panel of OPN and IL-6 discriminated the five-year survival group from the death group (H), the TNM stage I group from the healthy controls (I), and the metastasis group from the non-metastasis group (J). AUC, area under the curve. (K) A proposed model illustrating the modulatory role of stromal IL-6-induced neoplastic OPN in controlling tumor growth and metastasis. (* p <0.05; ** p <0.01; *** p <0.001; **** p <0.0001)

Article Snippet: The supernatant OPN and IL-6 concentrations and the plasma OPN and IL-6 levels in patients with HNC were assessed using the Human OPN ELISA kit (Boster, China) and human IL-6 ELISA kit (Boster, China).

Techniques: In Vivo, Over Expression, Clinical Proteomics, Injection, Activity Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: ADMET & DMPK

Article Title: Anti-inflammatory potential of plant-derived extracellular vesicles from Solanum nigrum L. integrated in gelatine-dopamine hydrogel on RAW 264.7 and MC3T3 cells

doi: 10.5599/admet.3149

Figure Lengend Snippet: Interleukin-6 (IL-6) concentration by (A) RAW 264.7 and (B) MC3T3 cells LPS induction and PDEV (2.5, 5 and 10 μg mL -1 ) treatment released from Gel-Dop hydrogel. Data presented as mean ± standard deviation (*** p < 0.001, **** p <0,0001)

Article Snippet: Other materials used in this study were Dulbecco’s modified eagle medium (DMEM) - high glucose, foetal bovine serum (FBS), Antibiotic-Antimycotic (ABAM), polyethylene glycol (PEG) 6000, trehalose (Sigma-Aldrich; Merck), phosphate-buffered saline (PBS), sodium periodate (NaIO 4 ) (Sigma-Aldrich), bovine serum albumin (BSA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT), 4′,6-diamidino-2-phenylindole (DAPI), dimethyl sulfoxide (DMSO), paraformaldehyde (PFA), kit BCA Assay (PierceTM BCA Protein Assay Kit, Thermo Scientific), PKH67 Green Fluorescent Cell Linker Kits (Sigma-Aldrich; Merck), and Mouse Interleukin-6 (IL-6) ELISA Kit (BT Lab, Cat. No. E0049Mo).

Techniques: Concentration Assay, Standard Deviation

Journal: BMC Pediatrics

Article Title: Placental determinants of fetal growth: identification of key factors in the insulin-like growth factor and cytokine systems using artificial neural networks

doi: 10.1186/1471-2431-8-24

Figure Lengend Snippet: Connectivity map clarifying the clusters of variables, and single relationships among variables. Mother_Age_Delivery: Chronological age of the mother at delivery (years); newborn_gender: Sex of the newborn; Gestational_age: Gestational age (weeks); Other_siblings: Other siblings (yes or No); N_siblings: Number of other siblings; N_abortion: Number of previous abortions; Pla_total_pro: Total placental protein content in lysates (mg/ml); PLA_IGF2: IGF-II concentration in placental lysates (ng/mg); PLA_BP2: IGFBP-2 concentration in placental lysates (ng/mg); PLA_IL6: IL-6 concentration in placental lysates (pg/mg); PL_TNF: TNF- concentration in placental lysates (ng/mg); mRNA_IGF1:IGF-I relative gene expression in placenta (A.U./18S, UBQ); mRNA_IGF2:IGF-II relative gene expression in placenta (A.U./18S, UBQ); mRna_BP1: IGFBP-1 relative gene expression in placenta (A.U./18S, UBQ); mRNA_BP2:IGFBP-2 relative gene expression in placenta (A.U./18S, UBQ); mRNA_IL6:IL-6 relative gene expression in placenta (A.U./18S, UBQ); Controls: newborns appropriate for gestational age; IUGR: Intra uterine growth retardation (FGR fetal growth restricted).

Article Snippet: IGF-I (Assay ID: Hs00153126-m1), IGF-II (Assay ID: Hs00171254-m1), IGFBP-1 (Assay ID: Hs00426258-m1), IGFBP-2 (Assay ID: Hs00167151-m1) and IL-6 (Assay ID: Hs00174131-m1) primers were used according to the manufacturer's instructions.

Techniques: Concentration Assay, Gene Expression