lncRNA FR215775 Regulates Th2 Differentiation in Murine Allergic Rhinitis.

To identify the effect of long noncoding RNA (lncRNA) FR215775 in regulating CD4+ T cells on murine models of allergic rhinitis (AR), the expression of lncRNA FR215775 in primary Th2 cells was detected through qRT-PCR. After knocking down the expression of lncRNA FR215775 via Sh-FR215775-Ads, Cell Counting Kit-8, cytometric bead array, and fluorescence-activated cell sorting were performed to determine its functions in vitro. Moreover, lncRNA FR215775-silencing or nonsilencing cells were injected intravenously into AR mice. Then, hematoxylin and eosin, Alcian blue-periodic acid Schiff, and toluidine blue staining were performed, and the levels of IL-2, IL-4, IL-5, IL-6, IL-10, IL-17A, IFN-γ, and TNF in the AR mice were also determined. We found that the expression of lncRNA FR215775 was specifically higher in the murine primary Th2 cells. After the knockdown of lncRNA FR215775, the proliferation of CD4+ T cells was inhibited, and the expressions of IL-4 and IL-5 in the cell culture supernatant were significantly decreased (P < 0.001), along with the percentage of Th2 cells (P < 0.05). The lncRNA FR215775-silencing AR group showed less serious allergic symptoms and a low level of ovalbumin-specific immunoglobulin E (P < 0.01). Meanwhile, the eosinophilia inflammation, goblet cell hyperplasia, and mast cell inflammation in the nasal mucosa all decreased, which indicated attenuated allergic inflammation in the lncRNA FR215775-silencing AR group. In addition, the Th2-related cytokines IL-4 and IL-5 were downregulated in the serum and nasal lavage fluid of this group (P < 0.01). In conclusion, lncRNA FR215775 may play a vital role in the function and differentiation of Th2 cells, which may encourage allergic inflammation. These results may provide significant insights into AR pathogenesis and offer new treatment targets for alleviating AR.

The Role of graRS in Regulating Virulence and Antimicrobial Resistance in Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a common cause of both community- and hospital-associated infections. The antibiotic resistance and virulence characteristics of MRSA are largely regulated by two-component signal transduction systems (TCS) including the graRS TCS. To make a relatively comprehensive insight into graRS TCS in MRSA, the bioinformatics analysis of dataset GSE26016 (a S. aureus HG001 WT strain vs. the ΔgraRS mutant) from Gene Expression Omnibus (GEO) database was performed, and a total of 563 differentially expressed genes (DEGs) were identified. GO analysis revealed that the DEGs were mainly enriched in the "de novo" IMP biosynthetic process, lysine biosynthetic process via diaminopimelate, and pathogenesis; and they were mainly enriched in purine metabolism, lysine biosynthesis, and monobactam biosynthesis in KEGG analysis. WGCNA suggested that the turquoise module was related to the blue module, and the genes in these two modules were associated with S. aureus virulence and infection. To investigate the role of graRS in bacterial virulence, a graRS knockout mutant (ΔgraRS) was constructed using MRSA USA500 2,395 strain as a parent strain. Compared to the wild-type strain, the USA500ΔgraRS showed reduced staphyloxanthin production, retarded coagulation, weaker hemolysis on blood agar plates, and a decreased biofilm formation. These altered phenotypes were restored by the complementation of a plasmid-expressed graRS. Meanwhile, an expression of the virulence-associated genes (coa, hla, hlb, agrA, and mgrA) was downregulated in the ΔgraRS mutant. Consistently, the A549 epithelial cells invasion of the ΔgraRS mutant was 4-fold lower than that of the USA500 wild-type strain. Moreover, on the Galleria mellonella infection model, the survival rate at day 5 post infection in the USA500ΔgraRS group (55%) was obviously higher than that in the USA500 group (20%), indicating graRS knockout leads to a decreased virulence in vivo. In addition, the deletion of the graRS in the MRSA USA500 strain resulted in its increased susceptibilities to ampicillin, oxacillin, vancomycin, and gentamicin. Our work suggests that the graRS TCS plays an important role in regulating S. aureus virulence in vitro and in vivo and modulate bacterial resistance to various antibiotics.

Hub Genes Identification in a Murine Model of Allergic Rhinitis Based on Bioinformatics Analysis.

This study aimed to identify allergic rhinitis (AR)-related hub genes and functionally enriched pathways in a murine model. Dataset GSE52804 (including three normal controls and three AR mice) was downloaded from Gene Expression Omnibus (GEO). Differentially expressed genes (DEGs) were identified. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and protein-protein interaction (PPI) analyses of DEGs were performed to identify the hub genes in AR. The DEGs were classified into different modules by using the weighted gene co-expression network analysis (WGCNA). Moreover, to verify the potential hub genes, nasal mucosa tissues were obtained from murine AR models (n = 5) and controls (n = 5), and qRT-PCR and Western blot were performed. In this study, a total of 634 DEGs were identified. They were significantly enriched in 14 GO terms, such as integral component of membrane, plasma membrane, and G-protein-coupled receptor signaling pathway. Meanwhile, there were eight terms of KEGG pathways significantly enriched, such as Olfactory transduction, Cytokine-cytokine receptor interaction, and TNF signaling pathway. The top 10 hub genes (Rtp1, Rps27a, Penk, Cxcl2, Gng8, Gng3, Cxcl1, Cxcr2, Ccl9, and Anxa1) were identified by the PPI network. DEGs were classified into seven modules by WGCNA. According to qRT-PCR validation of the five genes of interest (Rtp1, Rps27a, Penk, Cxcl2, and Anxa1), the expression level of Rtp1 mRNA was significantly decreased in the AR group compared with the control group, while there are enhanced Rps27a, Penk, Cxcl2, and Anxa1 mRNA expressions in the AR mice group compared with the control group. Western blot was also performed to further explore the expression of Anxa1 in the protein level, and the results showed a similar expression trend.

Diffusion Kurtosis Imaging and Intravoxel Incoherent Motion in Differentiating Nasal Malignancies.

OBJECTIVES/HYPOTHESIS: To evaluate the usefulness of diffusion kurtosis imaging (DKI) and intravoxel incoherent motion (IVIM) in the differentiation of sinonasal malignant tumors (SNMTs) with different histological types. STUDY DESIGN: Retrospective observational and diagnostic study. METHODS: Sixty-five patients with SNMTs who underwent DKI and IVIM were enrolled in this retrospective study, including 27 squamous cell carcinomas (SCCs), 13 olfactory neuroblastomas (ONBs), 14 malignant melanomas (MMs) and 11 lymphomas. The kurtosis (K) and diffusion coefficient (Dk) from DKI and the pure diffusion coefficient (D), pseudodiffusion coefficient (D*), perfusion fraction (f), and the product of D* and f (f∙D*) from IVIM were measured. Kruskal-Wallis and Dunn multiple comparison tests with Bonferroni correction, receiver operating characteristic curve, and logistic regression analyses were used for statistical analysis. RESULTS: Lymphomas demonstrated the highest K values but lowest Dk, D, D*, f, and f∙D* values among these four malignant tumors. ONBs exhibited high K values and MMs had highest D*, f, and f∙D* values. The cutoff value of ≤0.887 × 10-3 mm2 /sec for f∙D* provided a sensitivity, specificity, and an accuracy of 100%, 98.1%, and 98.5%, respectively, for differentiating lymphomas from the other three entities. The combination of f∙D* and D values showed a sensitivity of 92.9% and a specificity of 92.5% for the discrimination of MMs from ONBs and SCCs. The K value was useful for differentiating ONBs from SCCs, with a threshold value of 0.942 (sensitivity, 84.6%; specificity, 63.0%). CONCLUSIONS: The combined use of DKI and IVIM is helpful for differentiating among four histological types of SNMTs. LEVEL OF EVIDENCE: 3 Laryngoscope, 2019.

Effect of nicotine on Staphylococcus aureus biofilm formation and virulence factors.

Staphylococcus aureus is a common pathogen in chronic rhinosinusitis (CRS) patients, the pathogenesis of which involves the ability to form biofilms and produce various virulence factors. Tobacco smoke, another risk factor of CRS, facilitates S. aureus biofilm formation; however, the mechanisms involved are unclear. Here, we studied the effect of nicotine on S. aureus biofilm formation and the expression of virulence-related genes. S. aureus strains isolated from CRS patients and a USA300 strain were treated with nicotine or were untreated (control). Nicotine-treated S. aureus strains showed dose-dependent increases in biofilm formation, lower virulence, enhanced initial attachment, increased extracellular DNA release, and a higher autolysis rate, involving dysregulation of the accessory gene regulator (Agr) quorum-sensing system. Consequently, the expression of autolysis-related genes lytN and atlA, and the percentage of dead cells in biofilms was increased. However, the expression of virulence-related genes, including hla, hlb, pvl, nuc, ssp, spa, sigB, coa, and crtN was downregulated and there was reduced bacterial invasion of A549 human alveolar epithelial cells. The results of this study indicate that nicotine treatment enhances S. aureus biofilm formation by promoting initial attachment and extracellular DNA release but inhibits the virulence of this bacterium.

A minimally invasive endoscopic transnasal retropterygoid approach to the upper parapharyngeal space: anatomic studies and surgical implications.

BACKGROUND: Surgery remains the mainstay of treatment for lesions in the parapharyngeal space. However, gaining access to the parapharyngeal space is often challenging. In this study we aim to describe a minimally invasive technique of approaching the upper parapharyngeal space through an endoscopic transnasal retropterygoid approach, based on anatomic studies and surgeries. METHODS: Six fresh human cadaver heads were prepared for anatomic study at the Surgical Neuroanatomy Laboratory of the Center for Cranial Base Surgery within the Department of Neurological Surgery at the University of Pittsburgh School of Medicine. Three clinical cases seen in the Department of Otolaryngology, Eye, Ear, Nose, and Throat Hospital, Shanghai Medical College of Fudan University, were used to illustrate the technique and feasibility of this approach and to assess its indications, advantages, and drawbacks. RESULTS: The medial pterygoid plate is the primary landmark of the endoscopic transnasal retropterygoid approach to the upper parapharyngeal space. Access to the upper parapharyngeal space could be obtained by removing the mucosa on the medial pterygoid plate and the mucosa below the pharyngeal orifice of the Eustachian tube. The 3 patients in our study tolerated the procedure well and had no serious complications after surgery. CONCLUSION: The anatomic data and clinical cases in this study confirm that an endoscopic transnasal retropterygoid approach is a feasible and effective surgical treatment for selected tumors in the upper parapharyngeal space.

Nicotine Enhances Staphylococcus epidermidis Biofilm Formation by Altering the Bacterial Autolysis, Extracellular DNA Releasing, and Polysaccharide Intercellular Adhesin Production.

Staphylococcus epidermidis is a common bacterial colonizer of human skin and mucous membranes, yet it has emerged as an important nosocomial pathogen largely due to its ability to form biofilms. Tobacco smoke has been demonstrated as a contributor to various infection diseases by improving the biofilm formation of multiple bacterial species; however, the association between tobacco smoke and S. epidermidis biofilm is still unclear. In this study, we tested the effect of nicotine, one of the most active components of tobacco, on S. epidermidis biofilm formation, and we studied the underlying mechanisms. Our results showed that nicotine promoted the biofilm formation of S. epidermidis 1457 strain (SE1457) and enhanced its initial attachment to a polyethylene surface as well as polysaccharide intercellular adhesin (PIA) production. In addition, an increased extracellular DNA release and a higher autolysis rate of SE1457 was detected after nicotine treatment, which was consistent with the increased ratio of dead cells in nicotine-treated SE1457 biofilm observed with confocal laser-scanning microscopy. Furthermore, the effect of nicotine on several autolysis-related and biofilm-related gene knockout mutants of SE1457 was tested. It showed that in ΔsaeRS, ΔlytSR, and ΔsceD, nicotine induced increase in biofilm formation was similar to that in SE1457; but in ΔarlRS, ΔatlE, and ΔicaC, the effect was obviously impaired. Consistently, the increase of the bacterial autolysis rate in ΔarlRS and ΔatlE induced by nicotine was not as significant as that in SE1457. Meanwhile, the growth inhibition of nicotine on SE1457 was observed, and it was much less on ΔarlRS and restored by the arlRS complementation. The arlRS transcription in SE1457 was inhibited by nicotine during cultivation as indicated by a promoter reporter assay using green fluoresent protein. Taken together, our study indicates that nicotine improves S. epidermidis biofilm formation by promoting its initial attachment and intercellular accumulation; the arlRS, atlE, and ica genes mediating bacterial autolysis and PIA production play an important role in this process.

An In Vitro Model to Study the Effect of 5-Aminolevulinic Acid-mediated Photodynamic Therapy on Staphylococcus aureus Biofilm.

Staphylococcus aureus (S. aureus) is a common human pathogen, which causes pyogenic and systemic infections. S. aureus infections are difficult to eradicate not only due to the emergence of antibiotic-resistant strains but also its ability to form biofilms. Recently, photodynamic therapy (PDT) has been indicated as one of the potential treatments for controlling biofilm infections. However, further studies are required to improve our knowledge of its effect on bacterial biofilms, as well as the underlying mechanisms. This manuscript describes an in vitro model of PDT with 5-aminolevulinic acid (5-ALA), a precursor of the actual photosensitizer, protoporphyrin IX (PpIX). Briefly, mature S. aureus biofilms were incubated with ALA and then exposed to light. Subsequently, the antibacterial effect of ALA-PDT on S. aureus biofilm was quantified by calculating the colony forming units (CFUs) and visualized by viability fluorescent staining via confocal laser scanning microscopy (CLSM). Representative results demonstrated a strong antibacterial effect of ALA-PDT on S. aureus biofilms. This protocol is simple and can be used to develop an in vitro model to study the treatment of S. aureus biofilms with ALA-PDT. In the future, it could also be referenced in PDT studies utilizing other photosensitizers for different bacterial strains with minimal adjustments.

Conditioned medium from umbilical cord mesenchymal stem cells improves nasal mucosa damage by radiation.

OBJECTIVES: To explore therapeutic effects of conditioned medium from human umbilical cord mesenchymal stem cells (hUC-MSCs) on nasal mucosa radiation damage both in vivo and in vitro. RESULTS: The mucus cilia clearance time (7 and 30 days), degree of mucosal edema (7, 30, 90 and 180 days), cilia coverage (180 days) of concentrated conditioned medium group improved compared with radiotherapy control group. The proliferation and migration abilities of irradiated and non-irradiated nasal epithelial cells significantly increased after culture in bronchial epithelial cell growth medium (BEGM) containing 10% conditioned medium of hUC-MSCs compared to cells cultured in BEGM alone. CONCLUSIONS: Soluble factors secreted by hUC-MSCs may promote nasal epithelial cell proliferation and migration. Intranasal administration of hUC-MSC conditioned medium effectively repairs nasal mucosa radiation damage.

The kinase LRRK2 is differently expressed in chronic rhinosinusitis with and without nasal polyps.

BACKGROUND: Chronic rhinosinusitis (CRS), commonly divided into CRS with nasal polyps (CRSwNP) and without nasal polyps (CRSsNP) is an inflammatory disease which mechanism remain unclear. Leucine-rich repeat kinase 2 (LRRK2) has been proved to be a negative regulator of inflammation response while its role in pathogenesis of CRS has yet to be revealed. This research study was designed to investigate the relationship between the expression level and biologic role of LRRK2 in CRS. METHODS: Expression of LRRK2 mRNA and noncoding repressor of NFAT (NRON) were examined by qRT-PCR. Protein levels of LRRK2 were performed by western blot and immunohistochemistry. Nuclear factor of activated T cells (NFAT) nuclear translocation was analyzed by immunohistochemistry. Additionally, LRRK2 mRNA and NRON expression in response to specific inflammatory stimulation was measured in human nasal epithelia cells (HNECs). RESULTS: The expression of LRRK2 was increased in CRSsNP patients (p  <  0.05) and positively correlated with the expression levels of CD3 and Charot-Leyden crystal. Meanwhile, the NRON expression level is much lower in CRSsNP patients compared to both the control group and CRSwNP group (p  <  0.05). Marked enhanced NFAT nuclear localization was observed in CRSwNP groups compared with the CRSsNP and control group (p  <  0.0001). And the over-expression of LRRK2 was significantly regulated by lipopolysaccharide (LPS) in HNECs (p  <  0.05). Moreover, IL-17A can increase LRRK2 expression and suppress NRON expression in vitro and dexamethasone can rescue the NRON inhibition. CONCLUSION: LRRK2 and NRON may play different role in CRSsNP and CRSwNP. The molecular mechanisms identified here may aid in the design of novel therapeutic strategies to improve clinical outcomes.

The role of YKL40 in the pathogenesis of CRS with nasal polyps.

BACKGROUND: Many Chinese patients who experience chronic rhinosinusitis with nasal polyps (CRSwNP) have been shown to exhibit specifically enhanced TH1/TH17 responses and excessive neutrophil accumulation without demonstrating significant eosinophilia. These patients may be subject to different pathologies and therapies compared to Western patients. YKL40 can be produced by neutrophil and is associated with many inflammatory diseases, while its role in the pathogenesis of chronic rhinosinusitis (CRS) has yet to be determined. OBJECTIVE: The aim of this study was to investigate the relationship between the expression level and biologic role of YKL40 in CRS. METHODS: YKL40 expression was examined via quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), immunohistochemistry, and Western blot. Human nasal epithelia cells (HNECs) were isolated to detect YKL40 expression in response to specific inflammatory stimulation. RESULTS: YKL40 expression levels were significantly higher in NP patients compared to the turbinates of CRSsNP/CRSwNP and the control group and can be strongly activated by stimulation with IL-4 in vitro and suppressed by the other pro-inflammatory cytokines; lipopolysaccharide (LPS) and dexamethasone also caused significant decreases in YKL40 expression in HNECs. CONCLUSIONS: YKL40 may play a significant role in Chinese patients with CRSwNP. The molecular mechanisms identified here may aid in the design of new therapeutic strategies for improving the clinical outcomes of Chinese patients.

Microarray analysis of lncRNA and mRNA expression profiles in mice with allergic rhinitis.

OBJECTIVES: We aimed to identify the effect of lncRNAs in CD4+ T cells on Allergic rhinitis (AR). METHODS: The present study conducted a microarray to identify the expression profiles of lncRNA and mRNA in CD4+ T cells in both AR murine models and normal controls. And qRT-PCR was used to confirm the results. GO and KEGG enrichment analysis were used to show all related pathways and a co-expression network was conducted to find lncRNAs which have high correlation with these pathways. RESULTS: The results showed that the two groups contained a total of 158 deregulated lncRNAs, of which 110 were upregulated and 48 were downregulated. And positive regulation of calcium ion transport, B cell activation, chemokine-signaling pathways and calcium-signaling pathways may be involved in the development of T cells in AR pathology. Finally, we can find the differentially expressed mRNA in the pathways related to T cell differentiation correlated with many deregulated lncRNAs. CONCLUSIONS: The present study was the first to show the differential expression profiles of lncRNAs in the CD4+ T cells of an AR murine model, which may provide significant insights into AR pathogenesis and offer new treatment targets to alleviate it.

Effect of Zhu-yuan decoction in patients with chronic rhinosinusitis after functional endoscopic sinus surgery.

OBJECTIVE: To investigate the effectiveness and safety of Zhu-yuan decoction (ZYD) in patients after functional endoscopic sinus surgery (FESS). METHODS: A total of 85 patients were randomized into two groups: 44 were treated with intranasal corticosteroids (INC), and 41 were given Chinese herbal medicine (CHM). Patients with chronic rhinosinusitis (CRS) who underwent FESS were prospectively enrolled in the study. Before surgery, they were evaluated by visual analog scale (VAS), nasal endoscopy, computed tomography (CT), and routine blood test. After surgery, they were randomized to take ZYD or INC for 12 weeks and revaluated by VAS; nasal endoscopy at 4, 8, and 12 weeks; and CT at 12 weeks after surgery. RESULTS: In the both groups, VAS and endoscopy scores decreased significantly at 4, 8, and 12 weeks, and CT scores after treatment declined at 12 weeks compared with baseline scores. No significant differences were observed with regard to postoperative VAS, endoscopy, or CT scores between groups. ZYD, combined with surgery, can reduce VAS, nasal endoscopy, and CT scores and has the same efficacy and safety profile as INC in post-FESS management. No fatalities or major adverse events occurred in either group. CONCLUSION: Our findings suggest that ZYD has similar effects and safety profiles in patients after FESS compared with INC.

The effect of blocking Notch signaling by γ-secretase inhibitor on allergic rhinitis.

OBJECTIVE: This study aimed to investigate the effect of blocking Notch signaling by γ-secretase inhibitor (GSI) on allergic rhinitis. METHOD: GSI, N-[N-(3,5-difluorophenacetyl-L-alanyl)]-S-phenylglycine t-butylester (DAPT) was administered to ovalbumin-induced AR mice models intranasally. We observed symptoms of sneezing and nose rubbing. To detect the inflammatory state, the serum OVA-specific-IgE, IFN-γ, IL-4, and IL-5 were analyzed by ELISA, and Th cell cytokines in nasal mucosa were analyzed by RT-PCR, including T-bet, IFN-γ, GATA-3, IL-4, and IL-5. In addition, hematoxylin-eosin (HE) and periodic acid-Schiff (PAS) were applied for histopathological examination. As for the evaluation of Notch signaling, we analyzed the Notch-1, Notch signaling target Hes-1, and Hes-5 in mucosa by RT-PCR, besides, used western blotting and immunohistochemistry to assess NICD (Notch intracellular domain). RESULTS: The results showed that the DAPT ameliorated the development of AR and suppressed Th2 cytokine levels significantly, alleviating eosinophils infiltration and goblet cells metaplasia, suggesting that the GSI can regulate Th2 response and weaken airway inflammation in AR. CONCLUSION: Our findings provide evidence that blocking Notch signaling by GSI offers high value in treating AR.

5-aminolevulinic acid-mediated photodynamic therapy and its strain-dependent combined effect with antibiotics on Staphylococcus aureus biofilm.

Staphylococcus aureus (S. aureus) is hard to be eradicated, not only due to the emergence of antibiotic resistant strains but also because of its ability to form biofilm. Antibiotics are the major approach to treating biofilm infections, but their effects are unsatisfactory. One of the potential alternative treatments for controlling biofilm infections is photodynamic therapy (PDT), which requires the administration of photosensitizer, followed by light activation. 5-aminolevulinic acid (ALA), a natural photosensitizer prodrug, presents favorable characteristics, such as easy penetration and rapid clearance. These advantages enable ALA-based PDT (ALA-PDT) to be well-tolerated by patients and it can be repeatedly applied without cumulative toxicity or serious side effects. ALA-PDT has been proven to be an effective treatment for multidrug resistant pathogens; however, the study of its effect on S. aureus biofilm is limited. Here, we established our PDT system based on the utilization of ALA and a light-emitting diode, and we tested the effect of ALA-PDT on S. aureus biofilm as well as the combined effect of ALA-PDT and antibiotics on S. aureus biofilm. Our results showed that ALA-PDT has a strong antibacterial effect on S. aureus biofilm, which was confirmed by the confocal laser scanning microscope. We also found that lethal photosensitization occurred predominantly in the upper layer of the biofilm, while the residual live bacteria were located in the lower layer of the biofilm. In addition, the improved bactericidal effect was observed in the combined treatment group but in a strain-dependent manner. Our results suggest that ALA-PDT is a potential alternative approach for future clinical use to treat S. aureus biofilm-associated infections, and some patients may benefit from the combined treatment of ALA-PDT and antibiotics, but drug sensitivity testing should be performed in advance.

Role of NLRP3 Inflammasome in Eosinophilic and Non-eosinophilic Chronic Rhinosinusitis with Nasal Polyps.

The pathophysiologic mechanisms of human chronic rhinosinusitis with nasal polyps (CRSwNP) remain unclear. We aimed to elucidate expression and biologic role of NLRP3 inflammasome in CRSwNP. Immunohistochemistry (IHC) was conducted to assess NLRP3 immunolabeling, real-time polymerase chain reaction (PCR) was used for IL-9 and NLRP3, and caspase-1 level quantitation in CRSwNP and control subjects. In addition, enzyme-linked immunosorbent assay (ELISA) was employed for analyzing concentrations of IL-1ß and IL-18 in the homogenates prepared from tissue specimens. Moreover, human nasal epithelial cells (HNECs) were used to evaluate the effects of lipopolysaccharide (LPS) and glyburide on NLRP3 inflammasome signaling pathway. Results showed that NLRP3 and caspase-1 were overexpressed in CRSwNP, especially in eosinophilic CRSwNP (ECRSwNP). Interestingly, NLRP3 expression had close correlation to that of caspase-1. Concentrations of IL-1ß and IL-18 were elevated. NLRP3 inflammasome signaling pathway was augmented by LPS but suppressed by glyburide. In conclusion, NLRP3 inflammasome signaling pathway played a pro-inflammatory role in the pathogenesis of CRSwNP, especially in ECRSwNP. NLRP3 inflammasome signaling pathway was augmented by LPS, but suppressed by glyburide.

[In-vitro study of photodynamic therapy of antibiotic-resistant staphylococcus from patients with chronic rhinosinusitis].

OBJECTIVE: To evaluate the photodynamic therapy (PDT) against multi-antibiotic-resistant Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S.epidermidis) obtained from patients with chronic rhinosinusitis (CRS). METHODS: Forty-five CRS patients who had been given medical treatment but still needed endoscopic surgery were included in this study. The mucus from middle meatus was collected from these patients during surgery, followed by separation of S. aureus and S. epidermidis and drug sensitive test. The strains which could form biofilm were selected. Light emitting diode (LED) array with a major wavelength of (633±10) nm was used as light source and 5-Aminolevulinic acid (ALA) was used as photosensitizer in this PDT experiment. The safe range of LED dose and ALA concentration which were not toxic to bacteria by themselves were confirmed, and then did PDT experiment on S. aureus and S. epidermidis. The data of bacterial colony forming unit were transformed to lgCFU before statistical analysis.The Graph Pad Prism 5 software was used to analyzed the data. RESULTS: Thirteen S. aureus and 16 S. epidermidis were included in this experiment(from 45 patients), all of them were multi-antibiotic-resistant bacteria, and four of S. aureus and five of S. epidermidis could form biofilm in each group. In planktonic S. aureus experiment, the mean lgCFU was 8.32±0.31 in control group whereas the experiment group was 6.47±0.67 (t=9.01, P<0.01), and in planktonic S. epidermidis experiment the final data was 8.34±0.20 (control group) and 6.97±0.59 (experiment group) (t=8.84, P<0.01). In biofilm S. aureus experiment, the mean lgCFU was 8.68±0.05 (control group), 6.90±0.96(experiment group) (t=3.68, P<0.05); and in biofilm S. epidermidis experiment the data was 8.67±0.05 (control group), 7.29±0.61 (experiment group, t=5.07, P<0.01). CONCLUSION: Our results demonstrated that ALA-mediated PDT on multi-antibiotic-resistant S. aureus and S. epidermidis from CRS patients was effective in vitro. Additional work defining if the PDT treatment would damage the nasal mucosa and further checking the effectiveness of PDT in vivo is still needed.

Role of Suppressor of Cytokine Signaling 3 in the Immune Modulation of Mesenchymal Stromal Cells.

The underlying mechanisms of mesenchymal stromal cells (MSCs) on immune modulation to treat allergic diseases remain unclear. Here, we showed that the suppressor of cytokine signaling 3 (SOCS3) is an important immune modulator expressed by MSCs, which is significantly increased by interferon-γ (IFN-γ). In addition, we observed that SOCS3 is a crucial mediator of the anti-proliferative and functional effects of MSCs on T cells and B cells. The immune modulation of MSCs through SOCS3 is mediated by cell-cell contacts. Moreover, SOCS3 could serve as an indicator to predict the potential immune modulatory of MSCs derived from different donors. Furthermore, treatment with anti-SOCS3 Ab significantly decreased ovalbumin-specific antibodies and neutrophil infiltration in ovalbumin-induced allergic rhinitis (AR) mice. Our results suggest that SOCS3 serves as an immune modulator interfering with T cells and B cells, and SOCS3 may act as a predictive marker for immune modulatory of MSCs.