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: BioMed Research International

Article Title: MP Resulting in Autophagic Cell Death of Microglia through Zinc Changes against Spinal Cord Injury

doi: 10.1155/2016/6090316

Figure Lengend Snippet: MP reduced the activation of microglia in SCI rats. After 12 hours and 18 hours treatment, spinal cord was collected for histological analysis, ELISA and RT-PCR. (a) Immonoexpression of Iba1 (brown) in spinal cord at 12 h and 18 h. Scale bar = 200 μ m, 50 μ m; (b), (c) Spinal cord around lesion was subjected to examine the expression of iNOS, IL-6, IL-10, LC-3B and Beclin-1 by RT-PCR. Data were expressed as mean ± SD ( n = 5 for each group). ∗ P < 0.05, ∗∗ P < 0.01 versus normal group; # P < 0.05, ## P < 0.05 versus SCI group.

Article Snippet: IL-6, IL-10, and TNF- α (tumor necrosis factor- α ) (both Origene) were measured using respective ELISA kit according to the manufacturer's instructions and analyzed by microplate reader (Dynex Technology, Chantilly, VA, USA).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: BioMed Research International

Article Title: MP Resulting in Autophagic Cell Death of Microglia through Zinc Changes against Spinal Cord Injury

doi: 10.1155/2016/6090316

Figure Lengend Snippet: Effects of MP on microglia activation and secretion of inflammatory cytokines. (a) The expression of Iba-1 of microglia. After 12 hours of treatment, immunofluorescence staining was performed to assess expression of Iba-1 (Iba-1 green; DAPI blue); scale bar = 50 μ m. (b) After 12 hours of incubation, the expression of protein iNOS in microglia was assessed in LPS group, LPS + MP group, and normal group by western blot. (c) Cytokine levels in culture medium. After 12 hours of treatment, the culture media were collected and expression of IL-6, IL-10, and TNF- α was detected by ELISA kit. Data were expressed as mean ± SD ( n = 5 for each group). ∗ P < 0.05, ∗∗ P < 0.01 versus normal group; # P < 0.05 versus LPS group.

Article Snippet: IL-6, IL-10, and TNF- α (tumor necrosis factor- α ) (both Origene) were measured using respective ELISA kit according to the manufacturer's instructions and analyzed by microplate reader (Dynex Technology, Chantilly, VA, USA).

Techniques: Activation Assay, Expressing, Immunofluorescence, Staining, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay