Smell and taste dysfunction in patients infected with the Omicron variant of severe acute respiratory syndrome coronavirus-2.

BACKGROUND: Smell and taste dysfunctions (STD) are frequently observed in patients with coronavirus disease (COVID-19). OBJECTIVES: To investigate the clinical characteristics of STD in COVID-19 patients. MATERIAL AND METHODS: One-hundred six COVID-19 adult patients with the Omicron variant were enrolled. The clinical features of patients with and without STD were compared using questionnaires, laboratory tests, and imaging examinations. RESULTS: Of the 76 patients with smell and/or taste dysfunction, age (p = .002), vaccination time (p = .024), history of systemic diseases (p = .032), and smoking status (p = .044) were significantly different from those of the controls (n = 34). Fatigue (p = .001), headache (p = .004), myalgia (p = .047), and gastrointestinal discomfort (p = .001) were observed more frequently in these patients than in controls. The Hospital Anxiety and Depression Scale score of these patients was significantly higher than that of controls (p < .001). The taste visual assessment scale score of the STD group was significantly lower than that of the taste dysfunction group (p = .001), and perceptions of sour, sweet, and salty tastes were worse in the STD group than in the taste dysfunction group (p < .001). CONCLUSIONS AND SIGNIFICANCE: COVID-19 patients had similar changes in smell and/or taste dysfunctions and worse emotional states, possibly correlated with some factors, including age and vaccination time.

Clinical Features and Influencing Factors for the Prognosis of Patients With Sudden Deafness.

Backgrounds: Studies on risk factors influencing the prognosis of patients with sudden onset deafness are lacking. Methods: From March 2018 to March 2021, 500 patients, from the Tongde Hospital in Zhejiang Province, with sudden onset deafness were enrolled. We collected clinical information from the hospital medical records, including certain demographic characteristics, information related to sudden-onset deafness, and laboratory parameters. Univariate and multivariate analyses were performed to determine independent prognostic risk factors for patients with sudden deafness. Additionally, we also employed orthogonal partial least squares discriminant analysis (OPLS-DA) to analyze the data of these enrolled patients. Results: The baseline clinical characteristics of the enrolled patients were analyzed. Based on their prognoses, the included patients were divided into the overall effective and ineffective groups. Between these two groups, the univariate and multivariate analyses were performed. Age, type of hearing curve at the initial diagnosis, acute phase, and sudden deafness site were found to be independently associated with the prognoses of patients with sudden deafness (all P < 0.05). Through the OPLS-DA, the sudden deafness site was found to be an indicator with the highest predictive power. Conclusions: Age, type of hearing curve at the initial diagnosis, acute phase, and sudden deafness site were all independently correlated with the prognoses of patients with sudden deafness and, therefore, need to be emphasized.

Role and Function of Regulatory T Cell in Chronic Rhinosinusitis with Nasal Polyposis.

Chronic rhinosinusitis with nasal polyps (CRSwNP) is a subtype of chronic rhinosinusitis characterized by high edema in the stroma, albumin deposition, and formation of pseudocysts. The pathogenesis of CRSwNP is not yet fully understood. Regulatory T (Treg) cells are a subset of CD4+ T cells that play a suppressive immunoregulatory role in the process of CRSwNP. Recent studies have found that there was a significant reduction in Treg cells in polyp tissues, which leads to the onset of CRSwNP. An imbalance between Th17 and Treg cells can also aggravate inflammation toward the Th2 type. This review focuses on our understanding of the function and role of Treg cells and their regulatory factors and clinical significance in CRSwNP. We also summarize the current drug treatments for CRSwNP with Tregs as the potential therapeutic target, which will provide new ideas for the treatment of CRSwNP in the future.

SLC3A2 inhibits ferroptosis in laryngeal carcinoma via mTOR pathway.

OBJECTIVE: This study aimed to explore the mRNA and protein expression of SLC3A2 in laryngeal carcinoma cells and tissues, and functional regulatory mechanism of SLC3A2 in cell ferroptosis of laryngeal carcinoma. METHODS: We chose the key gene-SLC3A2 of DEGs from TCGA by bioinformatics analysis, and then we constructed stable knockdown of SLC3A2 in laryngeal carcinoma cells. MTT assay and clonogenic assay were used to determine cell viability and cell growth, respectively. The mRNA and protein expression were determined by RT-qPCR and western blotting, respectively. Xenograft tumor model was used to determine the role of SLC3A2 in tumor growth. RESULTS: The results of limma analysis recovered that 92 genes were involved in both upregulated DEGs and high risk of poor prognosis, whereas 36 genes were involved in both downregulated DEGs and low risk of poor prognosis. Pathway enrichment analysis indicated that mTOR signaling pathway and ferroptosis exerted a role in regulating these intersection genes. Moreover, SLC3A2 is a key gene in ferroptosis in laryngeal carcinoma. SLC3A2 is highly expressed in laryngeal carcinoma tissues and cells. Patients with high SLC3A2 expression exerted poor survival. SLC3A2 deficiency inhibited cell proliferation and foci formation. Furthermore, knockdown of SLC3A2 expression induced the efficacy of ferroptosis and suppressed ferroptosis related proteins expression. Mechanically, SLC3A2 deficiency facilitated ferroptosis through upregulating the expression of mTOR and P70S6K, whereas inhibited p-mTOR and p-P70S6K expression in laryngeal carcinoma cells. SLC3A2 deficiency inhibited tumorigenesis in nude mice. CONCLUSION: Our study suggests that SLC3A2 negatively regulates ferroptosis through mTOR pathway in laryngeal carcinoma.

MicroRNA-222 alleviates radiation-induced apoptosis by targeting BCL2L11 in cochlea hair cells.

Radiation-induced hair cell injury is detrimental for human health but the underlying mechanism is not clear. MicroRNAs (miRNAs) have critical roles in various types of cellular biological processes. The present study investigated the role of miR-222 in the regulation of ionizing radiation (IR)-induced cell injury in auditory cells and its underlying mechanism. Real-time PCR was performed to identify the expression profile of miR-222 in the cochlea hair cell line HEI-OC1 after IR exposure. miRNA mimics or inhibitor-mediated up- or down-regulation of indicated miRNA was applied to characterize the biological effects of miR-222 using MTT, apoptosis and DNA damage assay. Bioinformatics analyses and luciferase reporter assays were applied to identify an miRNA target gene. Our study confirmed that IR treatment significantly suppressed miR-222 levels in a dose-dependent manner. Up-regulation of miR-222 enhances cell viability and alleviated IR-induced apoptosis and DNA damage in HEI-OC1 cells. In addition, BCL-2-like protein 11 (BCL2L11) was validated as a direct target of miR-222. Overexpression of BCL2L11 abolished the protective effects of miR-222 in IR-treated HEI-OC1 cells. Moreover, miR-222 alleviated IR-induced apoptosis and DNA damage by directly targeting BCL2L11. The present study demonstrates that miR-222 exhibits protective effects against irradiation-induced cell injury by directly targeting BCL2L11 in cochlear cells.

Immune cell infiltration and related core genes expression characteristics in eosinophilic and non-eosinophilic chronic rhinosinusitis with nasal polyps.

Chronic rhinosinusitis with nasal polyps (CRSwNP) refers to chronic inflammation of the sinonasal mucosa. It can either be eosinophilic (ECRSwNP) or non-eosinophilic (non-ECRSwNP). However, immune cell infiltration in the microenvironment and pathogenesis of ECRSwNP and non-ECRSwNP are still unclear. The aim of the present study was to assess the immune cell infiltration and molecular mechanisms of ECRSwNP and non-ECRSwNP. In the present study, 22 immune cell types in ECRSwNP and non-ECRSwNP were investigated by CIBERSORT based on transcriptome data. The core gene related pathophysiology of CRSwNP was analyzed using Weighted Gene Correlation Network Analysis according to the phenotype of the infiltrated eosinophils and nasal polyps (NP). A total of four types of immune cells (mast cells, activated dendritic cells, M2 macrophages and activated natural killer cells) were demonstrated to have a direct and indirect correlation with eosinophilic infiltration in ECRSwNP. M1 macrophages and activated CD4+ memory T cells were correlated with major immune cell types in non-ECRSwNP. NP could affect the expression of 'olfactory receptor activity' and 'channel activity' genes to impair the olfactory signaling pathway and neuroactive ligand receptor pathway. 'Cell adhesion molecule binding', 'cytokine receptor binding' and 'glucocorticoid receptor binding' were significantly enriched in ECRSwNP, whereas epithelial cell injury, autophagy and the mTOR pathway (hsa04140 and hsa04150) may serve an important role in the pathogenesis of non-ECRSwNP. There were significantly different immune cell infiltration and related core genes expression characteristics between ECRSwNP and non-ECRSwNP. The results of the present study provide an improved basis for elucidation of the mechanism and treatment of CRSwNP.

FOXD3 inhibits cell proliferation, migration, and invasion in nasopharyngeal carcinoma through regulation of the PI3K-Akt pathway.

FOXD3 has been found previously to positively regulate miR-26b, a tumor inhibitor of nasopharyngeal carcinoma (NPC). However, FOXD3's precise function and associated mechanism of action in NPC have not yet been investigated. In this study, the expression of FOXD3 mRNA and protein was evaluated using RT-qPCR, western blotting, and immunohistochemistry. Protein levels involved in the phosphoinositide 3-kinase - protein kinase B (PI3K-Akt) pathway were assessed by western blot, and cell proliferation was determined by MTT and colony forming assays. Additionally, cell apoptosis was assessed by flow cytometric assay. Finally, the migration and invasion capabilities of the NPC cells were determined using wound healing and Transwell assays. We found that FOXD3 levels were relatively low in NPC tissue and cells, while an increase caused the inhibition of the PI3K-Akt pathway. Functional experiments found that overexpression of FOXD3 suppressed cell proliferation, migration, and invasion and enhanced cell apoptosis in NPC C6661 cells. IGF-1, an activator of the PI3K-Akt pathway, reversed the inhibitory effect of FOXD3. Furthermore, we found upregulation of the PI3K-Akt pathway and upregulation of the inhibitory effects of FOXD3 on C6661 cellular activities. In conclusion, FOXD3 negatively affected the PI3K-Akt pathway to restrain the processes involved in C6661 cell pathology. These findings further exposed the function and downstream axis of FOXD3 in NPC and displayed a promising new target for NPC therapy.

Long non-coding RNA LINC00460 promotes head and neck squamous cell carcinoma cell progression by sponging miR-612 to up-regulate AKT2.

LncRNAs (long noncoding RNAs) have been shown to be potentially critical regulators in head and neck squamous cell carcinoma (HNSCC). LncRNA LINC00460 (long intergenic non-protein coding RNA 460), an "oncogene", regulates progression of various tumors. However, the tumorigenic mechanism of LINC00460 on HNSCC is yet to be investigated. In the current study, we discovered that LINC00460 was relatively up-regulated in both HNSCC cancer tissues and cell lines, and predicted a poor prognosis in HNSCC patients. Gain- and loss-of functional studies established that over-expression of LINC00460 promoted cell proliferation, invasion and migration of HNSCC cells in vitro, while the promotion abilities were suppressed via knockdown of LINC00460. Our results identified miR-612 as a novel target of LINC00460, whose expression suggested a negative correlation with LINC00460 in HNSCC tissues and cell lines. LINC00460 increased the expression of serine/threonine kinase AKT2 via sponging miR-612. Rescue experiments indicated that LINC00460 could promote HNSCC progression partially through inhibition of miR-612. Subcutaneous xenotransplanted tumor model confirmed that interference of LINC00460 suppressed in vivo tumorigenic ability of HNSCC via down-regulation of AKT2. In conclusion, our findings clarified the biologic significance of LINC00460/miR-612/AKT2 axis in HNSCC progression and provided novel evidence that LINC00460 may be a new potential therapeutic target for HNSCC.

Amyloid ß precursor protein silencing attenuates epithelial­mesenchymal transition of nasopharyngeal carcinoma cells via inhibition of the MAPK pathway.

Advances in the treatment of nasopharyngeal carcinoma (NPC) have significantly improved the local control rate; however, distant metastasis remains a principal cause of mortality. Previous studies have demonstrated that the expression levels of amyloid ß precursor protein (APP) are increased in NPC. The present study aimed to investigate the association between APP and the development of NPC. In order to knockdown APP expression, an APP­small interfering RNA vector was synthesized and transfected into SUNE­1 cells. Cell Counting Kit­8 assay was performed to assess cell viability. The migratory and invasive abilities of SUNE­1 cells were examined by wound healing and Transwell assays, respectively. Reverse transcription­quantitative polymerase chain reaction and western blotting were performed to measure the mRNA and protein expression levels of APP, and additional factors involved in epithelial­mesenchymal transition (EMT) and in the mitogen­activated protein kinase (MAPK) signaling pathway. APP silencing significantly suppressed cell viability, migration and invasion. In addition, APP interference downregulated the expression levels of metastasis­associated 1, matrix metalloproteinase (MMP)­2 and MMP­9; however, knockdown of APP led to upregulation of tissue inhibitor of metalloproteinases 2 and inhibited EMT. The phosphorylation levels of p38, extracellular signal­-regulated kinases 1/2 and c­Jun N­terminal kinases 1/2 were decreased following downregulation of APP. The present results suggested that APP knockdown may significantly inhibit the development of NPC by suppressing cell viability, migration and invasion, and by inhibiting the EMT process via downregulation of the MAPK signaling pathway. Therefore, APP may facilitate the development of a novel gene therapy for the treatment of NPC.

Knockdown of serpin peptidase inhibitor clade C member 1 inhibits the growth of nasopharyngeal carcinoma cells.

Nasopharyngeal carcinoma (NPC) is a type of cancer originating in the nasopharynx. There are no NPC­specific treatments available at present. Serpin peptidase inhibitor clade C member 1 (SERPINC1) serves roles in anticoagulation and anti­inflammation. The aim of the present study was to investigate the role of SERPINC1 in the proliferation and apoptosis of NPC cells. Tumor and adjacent healthy tissue samples were collected from patients with NPC. Additionally, the SERPINC1 gene was silenced in the HNE3 cell line using short interfering RNA targeted against SERPINC1 (SERPINC1­siRNA). Cell viability was determined via a Cell Counting Kit­8 assay; furthermore, proliferation and apoptosis were investigated via flow cytometry. Western blotting and reverse transcription­quantitative polymerase chain reaction analysis were performed to determine the expression levels of protein and mRNA. It was revealed that the expression levels of SERPINC1 mRNA and protein were increased in NPC tumor tissues compared with in adjacent healthy tissues. The expression of SERPINC1 mRNA and protein in HNE3 cells decreased following SERPINC1­siRNA transfection. Furthermore, knockdown of SERPINC1 promoted apoptosis and inhibited proliferation. It was also demonstrated that silencing SERPINC1 upregulated the expression of B­cell lymphoma-2 (Bcl­2)­associated X protein and p53 mRNA and protein, and downregulated that of Bcl­2, survivin and cyclin D1. Downregulation of SERPINC1 reduced the phosphorylation of phosphatidylinositol 3­kinase (PI3K), protein kinase B (Akt) and mammalian target of rapamycin (mTOR). Thus, SERPINC1 knockdown may promote the apoptosis of HNE3 cells and inhibit proliferation via the suppression of the PI3K/Akt/mTOR signaling pathway.

Increased expression of MUC5AC and MUC5B promoting bacterial biofilm formation in chronic rhinosinusitis patients.

OBJECTIVE: Binding of bacteria to mucin and then colonizing is the first step of bacterial biofilm (BBF) formation in chronic rhinosinusitis (CRS) patients. Yet, information is sparse on how mucins effects on BBF formation. In this study, we aimed to investigate the relationship between mucin expression and the formation of BBF in CRS patients. METHODS: Sinus mucosa were harvested from 24 patients undergoing endoscopic surgery for CRS. The positive of BBF formation were detected by scanning electronic microscopy (SEM) and the expression of MUC1, MUC2, MUC5AC, and MUC5B were determined by immunohistochemistry (IHC). The difference expression of mucins were analyzed between the BBF positive and negative cohorts in CRS patients. RESULTS: MUC5AC and MUC5B are two major mucins in CRS mucosa with the former mainly restricted to the goblet cells of epithelium and the latter mainly restricted to the submucosal glands. Expression of MUC5AC and MUC5B in the sinus mucosa of BBF(+) CRS group was significantly higher than those in BBF(-) CRS group (p<0.05). For MUC1 and MUC2, no significant difference was found between the two groups (p>0.05). CONCLUSION: Our study indicated that increased expression of MUC5AC and MUC5B may play an important role in the pathogenesis of BBF formation.

Bacterial biofilms in chronic rhinosinusitis and their relationship with inflammation severity.

AIMS: To identify the presence of bacterial biofilms on mucosal specimens from chronic rhinosinusitis (CRS) patients, and evaluate their relationship with severity of CRS. METHODS: A prospective study of biofilms presence on 24 CRS patients compared with 12 controls was designed. The presence of biofilms was determined by scanning electron microscopy (SEM), and associations with the preoperative Lund-MacKay CT scores, Johansson endoscopic scores, and the history of ESS were assessed. RESULTS: Biofilms were found in 13/24 CRS patients (54.2%) but in only 1/12 controls (8.3%; P<0.01). CRS patients with and without biofilms had similar preoperative Lund-MacKay CT and Johansson endoscopic scores (P>0.05). Patients with revision ESS showed a tendency of higher biofilms incidence (5/7, 71.4%) than those undergoing their first procedure (8/17, 47.1%), but did not reach a significant difference (P>0.05). CONCLUSIONS: The higher incidence of biofilms in CRS patients suggests a role in the pathogenesis of CRS, but no correlation with severity of CRS.

[Relationship between bacterial biofilms and chronic rhinosinusitis by scanning electron microscopic study].

OBJECTIVE: To determine the presence and the morphological features of bacterial biofilms in surgical specimens of patients with chronic rhinosinusitis (CRS) compared with control patients without CRS by scanning electron microscopy (SEM), and to evaluate the role of biofilm on the pathogenesis of CRS. METHODS: Fifteen patients with CRS undergoing endoscopic sinus surgery and 11 control patients with fracture of nasal bone were enrolled in this study. Clinical information was recorded from each patient. All patients underwent a thorough otolaryngological examination, preoperative paranasal sinus computerized tomography (CT) scanning which were evaluated according to the Lund-Mckay CT scoring system. All the samples including uncinate process, ethmoid mucosa from CRS group and uncinate process, ethmoid bulla from control group were prepared using standard methods for SEM. The presence of bacterial biofilms on the samples of two groups was observed by SEM. Statistical analysis was performed using SPSS 13.0. Continuous data was analyzed by Student t test and dichotomous data was analyzed by chi² or Fisher exact test. P was considered to be significant at a level of 0.05. RESULTS: Nine (60.0%) of the 15 patients were found to have evidence of biofilms. In control group, only 1 (9.1%) of 11 patients had biofilm. The difference was statistical significant (chi² = 6.949, P < 0.01). All controls except one had healthy appearing cilia and goblet cells without biofilms. All the 16 CRS patients showed aberrant findings of the mucosal surface with variation in degrees of severity from disarrayed cilia to complete absence of cilia and goblet cells. Among them the typical morphologic feature such as water channels, 3-D structure, and matrix-embedding spherical or elliptical bodies were noted in 9 cases. Five samples including one case from control showed cilia aggregation. The preoperative CT scores of the CRS patients with biofilms (n = 9) were significantly higher than those without biofilms (n = 6, t = 2.14, P < 0.05). CONCLUSIONS: The typical morphologic feature of BF such as water channels, 3-D structure, and matrix-embedding spherical or elliptical bodies were noted in sinus mucosa of patients with CRS by the SEM. The positive rate of bacterial biofilms in CRS group was significantly higher compared to control group, which indicated bacterial biofilms might play an important role in the pathogenesis of CRS. Besides the typical bacterial biofilm features, cilia aggregation was found in five cases of CRS patients. We consider cilia aggregation can be regarded as one morphologic feature of bacterial biofilm in nasal mucosa, which needs further study. The presence of bacterial biofilms in CRS patients is associated with paranasal CT scores, which indicates that bacterial biofilm is correlated with the severity of CRS.