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The aim of this study was to investigate the role of lncRNA NEAT1 (nuclear enriched abundant transcript 1) in regulating Th2 cell differentiation. The overexpression vectors of NEAT1 and ITCH, and siRNA targeting NEAT1, EZH2 (enhancer of zeste homolog 2) and STAT6 (signal transducer and activator of transcription 6) were transfected into CD4+T cells. The mRNA expressions of ITCH and STAT6 were analyzed by qRT-PCR and western blotting. The levels of Th2 cytokines were detected by ELISA assay. RIP and ChIP assays were performed to analyze the association between NEAT1 and EZH2 as well as EZH2 and STAT6, respectively. Results showed that NEAT1 significantly repressed ITCH expression and increased STAT6 expression as well as the levels of IL-4, IL-5 and IL-13 in CD4+T cells. RIP and ChIP assays revealed that NEAT1 bound to EZH2 and EZH2 was recruited to the promoter region of ITCH in CD4+T cells. Silencing EZH2 significantly promoted STAT6 for ubiquitination. Furthermore, NEAT targeted STAT6 for ubiquitination and elevated levels of Th2 cytokines by regulating EZH2/ITCH axis. In conclusion, our data indicated that NEAT1 promotes Th2 cell differentiation through the EZH2/ITCH/STAT6 axis.

BACKGROUND Three-dimensional (3D) printed models have been shown to be promising in surgical training in rhinology. The objectives of this study were to develop a set of 3D-printed models including the pediatric and adult nasal cavity, and the postsurgical paranasal sinuses, and to assess the face and content validity in endoscopic training. METHODS The computed tomography (CT) data of a pediatric patient without nasal disorders and an adult patient with nasal septal deviation were selected to produce the models of the pediatric and adult nasal cavity, and the CT data of an adult patient who underwent endoscopic sinus surgery 4 months ago was chosen to create the paranasal sinus model. After the models were printed by our desktop-level 3D printer, 5 rhinologists used the 5-point Likert scales to evaluate the fidelity and utility. Additionally, a group of prespecified tasks were completed by the rhinologists and 5 residents respectively for supplementary content validation. The difference of time used in completing each task was analyzed by Mann-Whitney U test. RESULTS All the models were prototyped in 24 hours, and the total cost for each model was less than 100 CNY (15 USD). The overall scores for fidelity and usefulness in endoscopic training were above 4.0. The experts accomplished all tasks using significantly less time than the residents (all p < 0.05). CONCLUSION The models of nasal cavities and paranasal sinuses made by our desktop-level 3D printer are high-fidelity, low-cost, and useful in training basic endoscopic skills.

Staphylococcus aureus and Pseudomonas aeruginosa are primary pathogens in chronic rhinosinusitis (CRS), and the presence of S. aureus and P. aeruginosa biofilms has been associated with negative outcomes after surgery. This study investigated the inhibition effect of cetylpyridinium chloride (CPC)-quatsomes at low concentrations on both S. aureus and P. aeruginosa biofilms in vitro, as well as their toxicities towards cultured human airway epithelial (NuLi-1) cells. S. aureus ATCC 25923 and P. aeruginosa ATCC 15692 were used to establish biofilms. CPC-quatsome and CPC micelle solutions at concentrations of 0.01%, 0.025%, and 0.05% were prepared. AlamarBlue was used to test the viability of both planktonic S. aureus and P. aeruginosa and their biofilms after treatment for 5 min and 2 h, respectively. Confocal laser scanning microscopy (CLSM) was used to investigate the interactions between CPC-quatsomes and S. aureus and P. aeruginosa biofilms. A lactate dehydrogenase (LDH) assay was used to determine the toxicity of CPC-quatsomes on NuLi-1 cells. CPC-quatsome and CPC micelle solutions had significant inhibition effects at all tested concentrations on planktonic S. aureus and P. aeruginosa and their biofilms after 5-min exposure ( P < 0.05). In the CLSM study, different interactions between CPC-quatsomes and S. aureus or P. aeruginosa biofilms were observed. After 2-h treatment, the size of S. aureus biofilms decreased, while the number of dead bacteria increased in P. aeruginosa biofilms. Neither CPC-quatsomes nor CPC micelle solutions showed significant toxicity on NuLi-1 cell at all tested CPC concentrations ( P < 0.05). CPC-quatsomes at low concentrations inhibited S. aureus and P. aeruginosa in both planktonic form and biofilms. No adverse effects on NuLi-1 cells were observed, indicating their promising potential in the treatment of CRS. Impact statement Staphylococcus aureus and Pseudomonas aeruginosa biofilms are significant contributors to chronic rhinosinusitis (CRS), and are associated with poor prognosis. The killing effect of CPC-quatsomes on S. aureus biofilm at or above the CPC concentration of 0.5% (5 mg/mL) has been reported previously. This is the first study that showed the significant inhibition effect of CPC-quatsomes at low concentrations on both S. aureus and P. aeruginosa biofilms in vitro, and no adverse effects towards cultured human airway epithelial (NuLi-1) cells. In our study, CPC-quatsomes at concentrations of 0.01%, 0.025%, and 0.05% had significant inhibition effects on both planktonic and biofilms of S. aureus and P. aeruginosa. The result of this study indicates the promising potential of CPC-quatsome in the treatment of CRS.