Journal: EMBO Molecular Medicine

Article Title: A clinical and mechanistic study of topical borneol‐induced analgesia

doi: 10.15252/emmm.201607300

Figure Lengend Snippet: A–D Representative whole‐cell currents in HEK 293 cells expressing TRPV1 (A), ASIC3 (B), P2X2 (C), or P2X4 (D) in response to capsaicin (A), acidic pH (B), or ATP (C, D) in the absence of presence of borneol ( n = 5 for each channel). E The effect of different concentrations of borneol on the enzymatic activity of human COX‐2. The number of independent measurements is marked on top of each bar. F Averaged intracellular Ca 2+ increases in cultured mouse DRG neurons in response to consecutive applications of 200 μM borneol, 200 μM menthol, and 67 mM KCl. A total of 81 in 1,689 neurons from four mice were found to be borneol‐sensitive and were included in the analysis. G Representative intracellular Ca 2+ signals in HEK 293 cells expressing human TRPM8 (hTRPM8) in response to different concentrations of borneol. After each application of borneol, Ca 2+ ionophore ionomycin was applied to calibrate Ca 2+ response. RFU: relative fluorescence unit. H Dose–response curves of the borneol‐ or menthol‐induced increase in intracellular Ca 2+ in hTRPM8‐expressing HEK 293 cells. Smooth curves are fit to the Hill equation with an EC 50 of 65 μM and a Hill coefficient of 2.0 for borneol ( n = 15) and an EC 50 of 13 μM and a Hill coefficient of 2.0 for menthol ( n = 6 at concentrations of 0.1, 0.3, and 1 μM; n = 9 at concentration of 3 μM; n = 13 at concentrations of 10, 30, 100, and 300 μM). The data were normalized to ionomycin‐induced intracellular Ca 2+ increases. I Time course of menthol‐ and subsequently applied borneol‐induced whole‐cell currents in hTRPM8‐expressing HEK 293 cells ( n = 6). Data information: All the data are presented as the mean ± standard error of the mean (SEM).

Article Snippet: Intracellular calcium imaging of HEK 293 cells was performed using the FlexStation 3 microplate reader (Molecular Devices).

Techniques: Expressing, Activity Assay, Cell Culture, Fluorescence, Concentration Assay

Journal: EMBO Molecular Medicine

Article Title: A clinical and mechanistic study of topical borneol‐induced analgesia

doi: 10.15252/emmm.201607300

Figure Lengend Snippet: Representative intracellular Ca 2+ signals in HEK 293 cells transfected with empty vector in response to borneol and subsequent applied Ca 2+ ionophore ionomycin ( n = 6). Representative intracellular Ca 2+ signals in HEK 293 cells expressing hTRPM8 in response to different concentrations of menthol and the subsequently applied Ca 2+ ionophore ionomycin.

Article Snippet: Intracellular calcium imaging of HEK 293 cells was performed using the FlexStation 3 microplate reader (Molecular Devices).

Techniques: Transfection, Plasmid Preparation, Expressing

Journal: EMBO Molecular Medicine

Article Title: A clinical and mechanistic study of topical borneol‐induced analgesia

doi: 10.15252/emmm.201607300

Figure Lengend Snippet: Quantification of consecutively applied 100 μM menthol‐ and 600 μM borneol‐induced hTRPM8 currents. Currents were normalized to 100 μM menthol‐induced currents at +80 mV ( n = 6). Representative intracellular Ca 2+ signals in HEK 293 cells expressing mouse TRPM8 in response to different concentrations of borneol. Dose–response curves of borneol‐induced increase in intracellular Ca 2+ in mouse TRPM8‐expressing HEK 293 cells. The smooth curve is a fit to the Hill equation with an EC 50 of 116 μM ( n = 12). The data were normalized to ionomycin‐induced intracellular Ca 2+ increases. Time course of menthol‐ and subsequently applied borneol‐induced whole‐cell currents in mouse TRPM8‐expressing HEK 293 cells ( n = 5). Data information: All the data are presented as the mean ± standard error of the mean (SEM).

Article Snippet: Intracellular calcium imaging of HEK 293 cells was performed using the FlexStation 3 microplate reader (Molecular Devices).

Techniques: Expressing

Journal: Journal of Biological Chemistry

Article Title: dlk1/FA1 Regulates the Function of Human Bone Marrow Mesenchymal Stem Cells by Modulating Gene Expression of Pro-inflammatory Cytokines and Immune Response-related Factors

doi: 10.1074/jbc.m607530200

Figure Lengend Snippet: FIGURE 5. Dlk1 expression synergizes the effect of LPS-induced cytokines in hMSC cells and stimulates the B cell proliferation. A, control and hMSC-dlk1 cells were stimulated with 100 ng/ml LPS for 24 h. Total RNA was isolated and analyzed for the expression of the indicated genes by real time PCR. Expression of each target gene was normalized to -actin. Data are shown as means S.D. of three independent experiments. *, p 0.05; **, p 0.005 versus control cells (hMSC-TERT) induced with LPS. B, CD19-purified human peripheral blood B lymphocytes were cultured in CM obtained from hMSC-TERT (control cells) or hMSC-dlk1 cells as described under “Experimental Procedures” in the absence and the presence of 2.5 or 10 g/ml CD40L. Cell proliferation was measured with XTT-based assay. Stimulation in the cell proliferation was represented as fold induction over nonstimulated cells cultured in the corresponding CM. The bars are the means S.D. of five independent experiments. *, p 0.05; **, p 0.005 versus cultured cells in control/CM.

Article Snippet: Cells were then resuspended at 0.5 106/ml in RPMI (as a positive control), serum-free conditioned medium from hMSC-TERT (control/CM), or hMSC-dlk1 (dlk1/CM) supplemented with 10% FCS and stimulated with 20 ng/ml recombinant human IL-4 (R&D Systems, Abingdon, UK) and the indicated concentrations of recombinant human CD40L (R&D Systems) or left untreated (nonstimulated).

Techniques: Expressing, Control, Isolation, Real-time Polymerase Chain Reaction, Purification, Cell Culture, XTT Assay