IRF4 Knockdown Inhibits the Chronic Rhinosinusitis Without Nasal Polyps Development by Regulating NLRP3/Caspase-1/GSDMD-Mediated Pyroptosis.

Chronic rhinosinusitis without nasal polyps (CRSsNP) is a CRS phenotype. However, the mechanisms of CRSsNP remains unclear. Differentially expressed genes (DEGs) were obtained from the GSE36830 and GSE198950 datasets through the GEO2R tool. The six hub genes were screened by the protein-protein interaction (PPI) network analysis and Cytoscape software. Then we constructed the mouse models of CRS and verified the expression levels of hub genes by reverse transcription quantitative PCR (RT-qPCR). Hematoxylin-eosin (HE) staining was employed to observe pathological alterations in mouse tissues. Casepase-3 expression was detected by immunohistochemistry (IHC). The levels of TNF-α, IL-12, IL-6, IL-1ß, LDH, and IL-18 were evaluated using enzyme-linked immunosorbent assay (ELISA). Pyroptosis-related protein expressions were measured by western blotting. Cell counting kit-8 (CCK-8) and flow cytometry were performed to assess the proliferation and apoptosis of lipopolysaccharide (LPS)-induced NP69 cells. Six hub DEGs were identified. The expression levels of IRF4, IKZF1, and CD79A were obviously increased in CRSsNP, while those of ADH6, ADH1A, and LDHC were significantly decreased. IRF4 knockdown attenuated the pathologic features of CRSsNP. IRF4 knockdown reduced levels of the TNF-α, IL-12, IL-6 IL-1ß, LDH, and IL-18 as well as the proteins expression of Casepase-1, GSDMD, and NLRP3 both in vivo and in vitro, implying that inflammation and pyroptosis were inhibited. IRF4 knockdown hinders the development of CRSsNP by inhibiting the inflammatory response and NLRP3/Caspase-1/GSDMD-mediated pyroptosis, which offers novel promising treatment strategies for clinical intervention.

Toxicity of micro(nano)plastics with different size and surface charge on human nasal epithelial cells and rats via intranasal exposure.

Micro (nano)plastics (MNPs) have become emerging environmental contaminants, yet their toxicity and systemic effects via intranasal exposure remain unclear. This study investigated the in vitro toxicity of thirteen polystyrene MNPs with different surface functionalization (carboxylic (C-PS), amino (A-PS), and bare (PS)) and sizes (20-2000 nm) on human nasal epithelial cells (HNEpCs) at 10-1250 µg/mL as well as their in vivo toxicity to rats via intranasal administration at 125 µg/mL. The in vitro study showed that PS20, PS50, A-PS50, PS500, and A-PS500 significantly inhibited cell viability, which was dependent on particle concentration. A-PS induced higher cytotoxicity than C-PS and PS, and most MNPs inhibited cell proliferation after 24-h. Flow cytometry analysis suggested that PS induced cell apoptosis, while A-PS caused cell necrosis. MNPs were phagocytosed by HNEpCs and entered nucleus. The in vivo study showed that MNPs inhibited dietary behaviors of rats. Histological analysis indicated that PS20, PS200, and A-PS50 thinned out nasal mucosa. Immunohistochemical analysis revealed that exposure to PS20, PS200, and A-PS50 enhanced expression of transient receptor potential cation channel subfamily M (melastatin) member 8 (TRPM8). Systemic effects including hepatocyte cytoplasmic vacuolation and renal tubule dilatation were observed. The results suggested that nasal inhalation of MNPs may disturb energy metabolism and damage upper respiratory tract, liver, and kidneys.

Characteristics of pharyngeal paraesthesia symptoms in patients with obstructive sleep apnoea.

PURPOSE: This study aimed to investigate pharyngeal paraesthesia symptoms in patients with obstructive sleep apnoea (OSA). MATERIAL AND METHODS: Patients with snoring and suspected OSA as well as age-matched controls were recruited. All participants underwent nocturnal polysomnography (PSG) and pharyngeal paraesthesia assessment using the Glasgow-Edinburgh throat scale (GETS). The incidence and severity of pharyngeal paraesthesia symptoms were compared between the groups. RESULTS: A total of 280 patients who snored or were suspected of having OSA and 35 healthy, age-matched controls were recruited. The total pharyngeal paraesthesia symptom score was significantly higher in the OSA group than in the healthy group (12 [5, 23] vs. 3 [0, 9]; p < 0.001). The most frequent pharyngeal paraesthesia symptoms in the snore patients were Q7 (catarrh down the throat) and Q3 (discomfort/irritation in the throat), which are related to the irritability of the throat. The incidence of Q7 (OSA, 58% vs. controls, 14%; χ2 = 23.66; p < 0.001), Q3 (OSA, 46% vs. controls, 3%; χ2 = 23.07; p < 0.001), Q1 (feeling of something stuck in the throat; OSA, 33% vs. controls, 6%; χ2 = 11.00; p = 0.001), Q6 (swelling in the throat; OSA, 31% vs. controls, 0%; χ2 = 14.53; p < 0.001), Q9 (want to swallow all the time; OSA, 20% vs. controls, 6%; χ2 = 6.28; p = 0.012), Q5 (throat closing off; OSA, 24% vs. controls, 6%; χ2 = 6.16; p = 0.013), and Q2 (pain in the throat; OSA, 23% vs. controls, 6%; χ2 = 5.32; p = 0.021) was significantly higher in the OSA group than in the controls CONCLUSIONS: Patients with obstructive sleep apnoea have higher pharyngeal paraesthesia symptoms scores and tend to have irritated throats compared to healthy controls. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT03506178.

References Values for the Current Perception Threshold in the Pharynx Based on a Study of a Healthy Population.

OBJECTIVES: The application of the current perception threshold (CPT) in the diagnosis of pharyngeal sensory abnormalities has rarely been studied, and there is a lack of reference values for this application. This study established a normal reference range for CPT in the pharynx based on a study of a healthy population. METHODS: The CPT values of the palatoglossal arch, posterior 1/3 of the lingual body and hard palate were measured in 60 healthy individuals at 2000, 250, and 5 Hz. The influencing factors were analyzed, and reference values for the CPT were established. RESULTS: There was no correlation between the CPT value and gender. Age was only correlated at 250 Hz level in the hard palate. The CPT values of the palatoglossal arch were 324.95 ± 82.422 at 2000 Hz, 66.90 ± 38.622 at 250 Hz, and 13 ± 14.93 (7.83-22.75) at 5 Hz. The CPT values of the posterior 1/3 of the lingual body were 359.17 ± 76.299 at 2000 Hz, 86.92 ± 35.151 at 250 Hz, and 19 ± 15.73 (13.03-28.75) at 5 Hz. The CPT values of the hard palate were 157.5 ± 61.75 (124-185.75) at 2000 Hz, 57.63 ± 28.785 at 250 Hz, and 22 ± 25.73 (11.03-36.75) at 5 Hz. CONCLUSIONS: The CPT values of the pharynx in healthy people were not related to gender. The CPT values of the hard palate for 250 Hz stimulation were related to age, and there were no relationships between the CPT values and age for the other frequencies and loci. We established a normal reference range of CPT values in the pharynx from measurements obtained from healthy populations.

Current Perception Threshold Testing in Pharyngeal Paresthesia Patients with Depression or Anxiety.

PURPOSE: Satisfactory quantitative diagnostic approaches to pharyngeal paresthesia patients with depression or anxiety remain to be explored. This study investigated the plausibility of current perception threshold (CPT) testing in diagnosing pharyngeal paresthesia in patients with depression or anxiety. PATIENTS AND METHODS: A total of 41 patients with pharyngeal paresthesia with depression or anxiety were recruited as the study group. Additionally, 60 healthy volunteers constituted the control group. The CPT values associated with 5-, 250-, and 2000-Hz electrical stimulation frequencies were measured at the palatal lingual arch and posterior third of the lingual body (two sensory nerve distribution sites in the pharynx). The normal range of CPT values of the above three frequencies was analyzed. The differences in the CPT values for sensory nerves were compared. RESULTS: There were no significant differences in age and sex between the study and control groups. The CPT values of the pharynx at the two tested sites were not significantly correlated with age and gender. The CPT value of the study group was significantly lower than that of the control group in the palatal lingual arch and posterior third of the lingual body at an electrical stimulation of 5 Hz (p<0.05). No significant differences in the CPT values at other frequencies were found between the two groups. CONCLUSION: CPT testing is effective in determining pharyngeal paresthesia in patients with depression and anxiety. Paresthesia of the pharyngeal sensory nerve region is caused by damaged C fibers.

Quantitative Evaluation of the Function of the Sensory Nerve Fibers of the Palate in Patients With Obstructive Sleep Apnea.

STUDY OBJECTIVES: To quantitatively evaluate the functional integrity of sensory nerve fibers of the palate in patients with obstructive sleep apnea (OSA) using the Neurometer system. METHODS: A total of 32 patients with OSA and 18 healthy control patients were included in the study. All participants were selected based on medical history, physical examination, and nocturnal polysomnography (PSG) and divided into two groups. The palatal sensory status of participants was examined with a Neurometer current perception threshold (CPT) system. The system was used to deliver an electrical stimulus at three different frequencies (2,000 Hz, 250 Hz, and 5 Hz) by an investigator blinded to the PSG results. RESULTS: There were no significant differences in the CPT values of the hard palate between the patients with OSA and control patients at any of the three stimulation frequencies. The differences in the CPT values of the soft palate between these groups failed to show any statistical significance at 250 Hz and 5 Hz. However, the patients with OSA showed significantly higher CPT values of the soft palate at 2,000 Hz than the age-matched healthy control patients (256.56 ± 129.34 versus 372.13 ± 152.06; P = .009). CONCLUSIONS: Our study revealed an impairment of 2,000 Hz-related sensory nerve function of the soft palate among patients with OSA. The CPT test could be a useful tool for the quantitative and selective assessment of the sensory nerve function in patients with OSA. Additional research is required to evaluate the different types of sensory nerve dysfunctions among such patients. CLINICAL TRIAL REGISTRATION: Registry: ClinicalTrials.gov; Title: The Effects of Nasal Airflow on Upper Airway Dilator Muscles During Sleep; Identifier: NCT03506178; URL: https://clinicaltrials.gov/ct2/show/NCT03506178. CITATION: An Y, Li Y, Chang W, Gao F, Ding X, Xu W, Han D. Quantitative evaluation of the function of the sensory nerve fibers of the palate in patients with obstructive sleep apnea. J Clin Sleep Med. 2019;15(9):1347-1353.

The effects of nasal decongestion on obstructive sleep apnoea.

BACKGROUND: Many studies have indicated associations between impaired nasal breathing and sleep disorders. However, the precise nature of the relationship between nasal patency and sleep remains unclear. PURPOSE: We analysed the effects of nasal patency on sleep architecture and breath in nasal obstruction-predominant obstructive sleep apnoea (NO-OSA) patients by applying nasal decongestant. MATERIAL AND METHODS: A randomized, placebo-controlled double-blind crossover study was performed in OSA patients with chronic nasal obstruction and without obvious pharyngeal narrowing. All OSA patients (confirmed by polysomnography) were recruited and completed 2 overnight studies (randomly applying oxymetazoline or placebo). Data collected after oxymetazoline or placebo treatments were compared. The ClinicalTrials.gov identifier is NCT03506178. RESULTS: Compared with placebo, oxymetazoline resulted in significant increase in rapid eye movement sleep (p = 0.027) and reduction of stage 1 sleep (p = 0.004), as well as arousal index (p = 0.002). Moreover, great improvements in apnoea/hypopnea index (AHI) were observed (p < 0.001); AHI in the supine position was significantly reduced (p = 0.001). Oxygen saturation during sleep was increased significantly [mean oxygen saturation (p = 0.005) and lowest oxygen saturation (p = 0.024)]. Oxygen desaturation index was significantly reduced (p < 0.001). CONCLUSIONS: Improving nasal patency by decongestant could improve sleep quality, AHI, and oxygen saturation level during sleep.

Analysis of epidermal growth factor signaling in nasal mucosa epithelial cell proliferation involved in chronic rhinosinusitis.

BACKGROUND: Aberrant epithelial repair has been observed in chronic rhinosinusitis (CRS) patients; however, the mechanism of epithelial cell repair regulation is unclear. Epidermal growth factor (EGF) plays an important role in regulating epithelial cell repair in lower airway and may be a critical factor in the remodeling processes of CRS. The objective of our research is to evaluate the differences between CRS and normal subjects and between chronic rhinosinusitis without nasal polys (CRSsNP) and chronic rhinosinusitis with nasal polys (CRSwNP) in the regulation of EGF pathways and the regulating proliferative position of classic Ras/Raf/MEK/ERK pathways. METHODS: We evaluated the proliferation rates of ethmoidal mucosal cells before and after stimulation with EGF, epidermal growth factor receptor (EGFR) kinase inhibitor AG1478, and extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor PD98059 using MTT assays. We also analyzed the sinonasal epithelial cells collected from control subjects and patients with CRS subtypes CRSsNP and CRSwNP for the expression of ERK1/2, phosphorylated ERK1/2, P21, P15, and P27 using western blotting analyses. RESULTS: The proliferation rates of sinonasal epithelial cells before and after EGF stimulation were lower in CRS patients than in the controls. AG1478 or PD98059 inhibitor treatment of control epithelial cells did not result in a significant difference in proliferation. Although, AG1478 and PD98059 inhibited the proliferation of CRS cells, the degree of proliferation inhibition was markedly different in CRSsNP. AG1478 suppressed the proliferation of CRSwNP epithelial cells, whereas PD98059 had no effect. The ratio of ERK1/2 phosphorylation in CRS cells was lower than that of the control cells. Cyclin-dependent kinase inhibitors were highly expressed in CRS cells compared with that of control cells. ERK1/2 and P27 showed differential expression in CRSsNP and CRSwNP. CONCLUSIONS: Differences existed in EGF pathways in CRS patients and normal subjects as well as in CRSsNP and CRSwNP. Classical Ras/Raf/MEK/ERK pathway may assume absolute superiority in control cells. Ras/Raf/MEK/ERK classical pathway and other pathways might be active at the same time to stimulate epithelial cell proliferation in CRSsNP. The function of Ras/Raf/MEK/ERK classical pathway was weaker in CRSwNP than in CRSsNP and when the classical pathway was blocked in CRSwNP, some other pathway could have completely compensated the proliferation induced by the Ras/Raf/MEK/ERK pathway.

Analysis of transforming growth factor ß signaling in chronic rhinosinusitis.

BACKGROUND: It has been reported that there is a significant difference in the local tissue concentration of transforming growth factor (TGF)-ß1 between chronic rhinosinusitis without nasal polyps (CRSsNP) and chronic rhinosinusitis without nasal polyps (CRSwNP) patients. TGF-ß has been reported to play an important role in regulating epithelial cell repair in lower airway remodeling and may be a critical factor involved in the remodeling process of chronic rhinosinusitis (CRS). METHODS: Ethmoidal mucosal samples collected from CRS and healthy control patients were analyzed for TGF-ß1, TGF-ß receptor I, TGF-ß receptor II, Smad3, phospho-Smad3, Smad7, and Smad anchor for receptor activation by Western blotting analysis. The proliferation of sinonasal epithelial cells at baseline and after TGF-ß1 and/or EGF stimulation was evaluated by the MTT assay. RESULTS: In CRSsNP, TGF-ß1, TGF-ß receptor I, TGF-ß receptor II, and Smad3 protein levels were significantly higher than controls. In CRSwNP, TGF-ß1, Smad3, and pSmad3 protein levels were significantly lower than controls. Smad7 protein was significantly higher in CRS than controls. In vitro experiments demonstrated that the baseline proliferation levels of sinonasal epithelial cells were lower in CRS than controls. CONCLUSIONS: CRSwNP is characterized by a lower level of TGF-signaling compared with the control. In CRSsNP, although the upstream signaling of TGF-ß was enhanced, the high Smad7 protein expression may restrain the downstream signaling components (e.g., pSmad3) and the TGF-ß antiproliferative effect on sinonasal epithelium. The difference in the local tissue concentration of TGF-ß1 between CRSsNP and CRSwNP patients did not result in significant differences in epithelial proliferation.