In the present study, we investigated whether apigenin significantly affects tumor necrosis factor-α (TNF-α)-induced production and gene expression of MUC5AC mucin in airway epithelial cells. Confluent NCI-H292 cells were pretreated with apigenin for 30 min and then stimulated with TNF-α for 24 h or the indicated periods. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Apigenin significantly inhibited MUC5AC mucin production and down-regulated MUC5AC gene expression induced by TNF-α in NCI-H292 cells. To elucidate the action mechanism of apigenin, effect of apigenin on TNF-α-induced nuclear factor kappa B (NF-κB) signaling pathway was also investigated by western blot analysis. Apigenin inhibited NF-κB activation induced by TNF-α. Inhibition of inhibitory kappa B kinase (IKK) by apigenin led to the suppression of inhibitory kappa B alpha (IκBα) phosphorylation and degradation, p65 nuclear translocation. This, in turn, led to the down-regulation of MUC5AC protein production in NCI-H292 cells. Apigenin also has an influence on upstream signaling of IKK because it inhibited the expression of adaptor protein, receptor interacting protein 1 (RIP1). These results suggest that apigenin can regulate the production and gene expression of mucin through regulating NF-κB signaling pathway in airway epithelial cells.

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Cricothyroid (CT) approach for vocal fold injection (VFI) has advantages of a low complication rate, suitability for in-office practice, and good patient compliance. However, it requires a high level of experience and a steep learning curve due to invisibility of needle. Recently, real-time light-guided VFI (RL-VFI) was developed for safe and precise injection into laryngeal structures under light guidance. Herein, we describe the development of a simulation-based training (SBT) program using RL-VFI for CT approach and report its preliminary application with in-training otolaryngologists. Methods: The workshop comprised 3 sessions: mini-lectures, and two hands-on training courses of conventional VFI and RL-VFI. Excised canine larynges and the device for RL-VFI were prepared for hands-on courses. Comfort levels for VFI was evaluated using visual analogue scale after each session. Trainees were requested to identify the needle tip on the target point lateral to vocal process. The time (s) to reach the target point was measured in all procedures. After workshop, all participants filled out questionnaires regarding their future preference for conventional VFI and RL-VFI. Results: Eleven otolaryngology residents participated in the study. The mean comfort levels were 1.7 ± 1.6, 5.5 ± 2.6, 4.8 ± 1.7, and 7.5 ± 1.6 for pre-workshop, post-lecture, post-conventional VFI, and post-RL-VFI ( P < .001). The mean time (s) to reach the target point were 146.4 ± 90.1 and 42.7 ± 40.5 for conventional VFI and RL-VFI ( P = .004). The mean preference scores were 4.2 ± 1.3 and 8.7 ± 1.3 for conventional VFI and RL-VFI ( P = .004). Conclusion: SBT program using RL-VFI might improve the comfort levels of trainees for VFI with CT approach. It would be helpful for trainees to practice VFI before trying it on actual patients.

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