Journal: Journal of ethnopharmacology

Article Title: Antiviral effect of fufang yinhua jiedu (FFYH) granules against influenza A virus through regulating the inflammatory responses by TLR7/MyD88 signaling pathway.

doi: 10.1016/j.jep.2021.114063

Figure Lengend Snippet: Fig. 7. In vivo inhibitory effect of FFYH on the inflammatory factors, TNF-α, IL-6, IFN-γ, IP10, and IL-1β caused by influenza a virus (H1N1). The concentration of inflammatory factors in mice sera were quantified with ELISA assays. ##p < 0.05, ###p < 0.01, compared with normal control group; *p < 0.05, **p < 0.01, compared with the virus-infected group (n = 6).

Article Snippet: ELISA kit for examining TNF-α (EK0529), IL-6 (EK0411), IFN-γ(EK0375), IP-10 (EK0736) and IL-1β (EK0394) were obtained from Boster (Wuhan, China).

Techniques: In Vivo, Virus, Concentration Assay, Enzyme-linked Immunosorbent Assay, Control, Infection

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: Frontiers in Cell and Developmental Biology

Article Title: lncRNA Gm16410 Mediates PM 2 . 5 -Induced Macrophage Activation via PI3K/AKT Pathway

doi: 10.3389/fcell.2021.618045

Figure Lengend Snippet: Macrophage activation caused inflammation of lung tissue in response to PM 2 . 5 . (A) Immunohistochemistry detection of the macrophage marker F4/80 in mouse lung tissues. Scale bar = 10 μm. (B) Pathological changes in the lung tissues. Scale bar = 10 μm. (C , D) Western blot analysis of TNF-α and IL-1β levels in the lung tissues. The plot gives a quantitative gray-scale analysis of the blot ( n = 3). (E) ELISA analysis of IL-6 in serum ( n = 5). (F) Real-time quantitative PCR analysis of IL-6, TNF-α, and IL-1β in the lung ( n = 8). (G , H) Western blot analysis of the P-PI3K, P-AKT, and NF-κB in mouse lung tissues. The plot gives a quantitative gray-scale analysis of the blot ( n = 3). (I) Immunohistochemistry detection of P-AKT and NF-κB in mouse lung tissues. Scale bar = 10 μm. Data are the means ± SEMs from at least three independent experiments, each performed in triplicate. * P < 0.5 and ** P < 0.01—levels of significance that are different from the control group at the levels, respectively.

Article Snippet: Mouse IL-6 ELISA kit was purchased from Boster (Wuhan, China).

Techniques: Activation Assay, Immunohistochemistry, Marker, Western Blot, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Control

Journal: BioMed Research International

Article Title: The Role of IL-6RA in UHMWPE Promotes Proliferation in Fibro-Like Synovial Cells

doi: 10.1155/2018/3928915

Figure Lengend Snippet: UHMWPE increased the expression of mIL-6R protein. FLS cells were incubated with UHMWPE (0, 0.01, 0.1, and 1g/L) for 7 d. The expression of IL-6R protein was detected by western blotting assay.

Article Snippet: The primary antibodies included caspase-3(Abcam, USA; ab4051), cleaved-caspase-3 (Cell Signaling Technology, USA; #9661), Bax (Abcam, USA; ab182733), Bcl-2 (Abcam, USA; ab692), IL-6R (Boster, China; A01425-1), β -actin (Bioss, China; bs-0061R), and GAPDH (Abcam, USA; ab8245) at 1:500 dilutions.

Techniques: Expressing, Incubation, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Complement 5a‐mediated trophoblasts dysfunction is involved in the development of pre‐eclampsia

doi: 10.1111/jcmm.13466

Figure Lengend Snippet: Expression of angiogenesis‐related factors in human placenta. ( A ) Relative mRNA levels of anti‐angiogenic factors ( IL ‐1β, TNF ‐α, IL ‐6, MCP ‐1 and sF lt1) and pro‐angiogenic ( PIGF , IL ‐10) in normal and pre‐eclamptic placentas. Data are represented as mean ± S.E.M. N = 4–6 in each group. * P < 0.05, ** P < 0.01, *** P < 0.001. ( B ) Immunofluorescence analysis of representative angiogenesis‐related factors (red) and Cy7 (green) in normal and PE placentas. Nuclei were counterstained with DAPI (blue). Scale bar: 60 μm.

Article Snippet: The human placenta slices were blocked in 10% normal goat serum for 30 min., then incubated with primary antibodies against CD31 (Santa Cruz), CD11b (BD Biosciences, San Jose, CA, USA), C5a (Comp Tech, A221), C5aR (Biolegend), cytokeratin 7 (Cy7, Santa Cruz), PIGF (Proteintech group, Wuhan, Hubei, China), sFlt1 (Life Technologies, Waltham, MA, USA), IL‐1β (Boster), IL‐6 (Boster) and MCP‐1 (Boster, Wuhan, Hubei, China) at 4°C overnight.

Techniques: Expressing, Immunofluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Complement 5a‐mediated trophoblasts dysfunction is involved in the development of pre‐eclampsia

doi: 10.1111/jcmm.13466

Figure Lengend Snippet: Trophoblasts stimulated with C5a display an anti‐angiogenic phenotype. ( A ) HTR ‐8/ SV neo cells treated with C5a showed a polarization towards an anti‐angiogenic phenotype with significantly increased mRNA levels of IL ‐1β, TNF ‐α, IL ‐6, MCP ‐1, sF lt1 and decreased mRNA level of PIGF and IL ‐10. The respective mRNA was normalized to β‐actin housekeeping gene. Data are represented as mean ± S.E.M . N = 3 in each group * P < 0.05, ** P < 0.01. ( B ) Immunofluorescence staining for sF lt1 and PIGF expression in HTR ‐8/ SV neo cells in the presence of C5a or PBS ( CON ). Scale bar: 60 μm.

Article Snippet: The human placenta slices were blocked in 10% normal goat serum for 30 min., then incubated with primary antibodies against CD31 (Santa Cruz), CD11b (BD Biosciences, San Jose, CA, USA), C5a (Comp Tech, A221), C5aR (Biolegend), cytokeratin 7 (Cy7, Santa Cruz), PIGF (Proteintech group, Wuhan, Hubei, China), sFlt1 (Life Technologies, Waltham, MA, USA), IL‐1β (Boster), IL‐6 (Boster) and MCP‐1 (Boster, Wuhan, Hubei, China) at 4°C overnight.

Techniques: Immunofluorescence, Staining, Expressing