Journal: Frontiers in endocrinology

Article Title: Excessive Iodine Promotes Pyroptosis of Thyroid Follicular Epithelial Cells in Hashimoto's Thyroiditis Through the ROS-NF-κB-NLRP3 Pathway.

doi: 10.3389/fendo.2019.00778

Figure Lengend Snippet: FIGURE 1 | Aberrant pyroptosis occurs in the thyroid tissues of HT patients and is induced by excessive iodine in vitro. (A,B) Gasdermin D (GSDMD) protein levels in the thyroid tissue of HT patients (n = 20) and controls (n = 10) were analyzed by immunoblots. “Control” indicates tissues from patients with nodular goiter of the thyroid. Representative immunoblotting results and quantification of GSDMD are shown. (C,D) Representative results of GSDMD immunohistochemical staining in HT tissues (n = 20) and control tissues (n = 10) are shown. Brown regions represent positive expression (original magnification, ×200; scale bars, 100 µm). (E–G) Nthy-ori3-1 cells were harvested after treatment with a gradient of concentrations of sodium iodide (NaI) for 24 h. The images presented are immunoblots probed for GSDMD (Hsp60 served as the loading control). All statistical results shown are representative of three replicates. (H) The cell viability of Nthy-ori 3-1 cells was assessed by CCK-8 assays after NaI treatment for 24 h. All statistical results shown are representative of three replicates. Significant differences and P-values were calculated by unpaired t-tests or one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Thyroid sections were blocked with 2% bovine serum albumin in PBS for 30min and then incubated with mouse anti-human GSDMD or mouse antihuman IL-1β antibodies (Santa Cruz, NJ, USA) at 4◦C overnight.

Techniques: In Vitro, Western Blot, Control, Immunohistochemical staining, Staining, Expressing, CCK-8 Assay

Journal: Frontiers in endocrinology

Article Title: Excessive Iodine Promotes Pyroptosis of Thyroid Follicular Epithelial Cells in Hashimoto's Thyroiditis Through the ROS-NF-κB-NLRP3 Pathway.

doi: 10.3389/fendo.2019.00778

Figure Lengend Snippet: FIGURE 3 | The NLRP3 inflammasome participates in excessive iodine-induced pyroptosis in TFCs. (A,B) The NLRP3 expression levels were measured by immunoblots when Nthy-ori 3-1 cells were treated with NaI with or without NAC (10 mM) or IKK-16 (2 µM) for 24 h. (C,D) Verification of the silencing efficiency of NLRP3 by siRNA in Nthy-ori3-1 cells is shown by immunoblots; NC indicates the negative control. (E,F) The protein levels of GSDMD were detected by immunoblots after transfection of siNLRP3 in NaI-treated Nthy-ori 3-1 cells. All statistical results shown are representative of three replicates. Significant differences and P-values were calculated by one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Thyroid sections were blocked with 2% bovine serum albumin in PBS for 30min and then incubated with mouse anti-human GSDMD or mouse antihuman IL-1β antibodies (Santa Cruz, NJ, USA) at 4◦C overnight.

Techniques: Expressing, Western Blot, Negative Control, Transfection

Journal: Frontiers in endocrinology

Article Title: Excessive Iodine Promotes Pyroptosis of Thyroid Follicular Epithelial Cells in Hashimoto's Thyroiditis Through the ROS-NF-κB-NLRP3 Pathway.

doi: 10.3389/fendo.2019.00778

Figure Lengend Snippet: FIGURE 5 | Proposed model of TFC pyroptosis in excessive iodine-promoted HT. Excessive iodine enters TFCs and induces the production of ROS, which activates NF-κB signaling and increases GSDMD-FL. In addition, the NLRP3 inflammasome is activated by NF-κB signaling, leading to GSDMD-FL cleavage into GSDMD-N, which triggers pore formation in the membrane and the release of mature IL-1β from cells, causing a sterile inflammatory response and further contributing to pyroptotic cell death and the subsequent promotion of HT.

Article Snippet: Thyroid sections were blocked with 2% bovine serum albumin in PBS for 30min and then incubated with mouse anti-human GSDMD or mouse antihuman IL-1β antibodies (Santa Cruz, NJ, USA) at 4◦C overnight.

Techniques: Membrane, Sterility

Journal: Pain medicine (Malden, Mass.)

Article Title: Neonatal bee venom exposure induces sensory modality-specific enhancement of nociceptive response in adult rats.

doi: 10.1111/pme.12296

Figure Lengend Snippet: Figure 1 A flowchart illustrating the experimental design.

Article Snippet: After being blocked in 5% normal goat serum in PBS containing 0.3% Triton X-100 for 30 minutes, sections were incubated overnight at 4°C with primary antibodies to NGF (1:200; rabbit anti-rat, Santa Cruz Biotechnology, Santa Cruz, CA, USA), TrkA receptor (1:200; rabbit antirat, Santa Cruz), BDNF (1:200; rabbit anti-rat, Santa Cruz), TrkB receptor (1:200; rabbit anti-rat, Santa Cruz), IL-1β (1:200; rabbit anti-rat, Santa Cruz), or COX-2 (1:200; goat anti-rat, Santa Cruz).

Techniques:

Journal: Pain medicine (Malden, Mass.)

Article Title: Neonatal bee venom exposure induces sensory modality-specific enhancement of nociceptive response in adult rats.

doi: 10.1111/pme.12296

Figure Lengend Snippet: Figure 8 Effects of neonatal inflammation on the spinal expression of IL-1β and COX-2 in adult rats. Compared with adult rats without neonatal inflammation, rats receiving the first BV injection on P21 and beyond showed up-regulated expression of IL-1β in the spinal cord dorsal horn. Scale bar: 200 μm. IOD = integral optical density. Sal-BV = rats receiving saline injection neonatally and then exposed to BV as adults. BV-BV = rats receiving BV neonatally and then again exposed to BV as adults. * P < 0.05, as compared with the Sal-BV group.

Article Snippet: After being blocked in 5% normal goat serum in PBS containing 0.3% Triton X-100 for 30 minutes, sections were incubated overnight at 4°C with primary antibodies to NGF (1:200; rabbit anti-rat, Santa Cruz Biotechnology, Santa Cruz, CA, USA), TrkA receptor (1:200; rabbit antirat, Santa Cruz), BDNF (1:200; rabbit anti-rat, Santa Cruz), TrkB receptor (1:200; rabbit anti-rat, Santa Cruz), IL-1β (1:200; rabbit anti-rat, Santa Cruz), or COX-2 (1:200; goat anti-rat, Santa Cruz).

Techniques: Expressing, Injection, Saline