OBJECTIVE To explore expression of IL-13 in eosinophilic chronic rhinosinusitis with nasal polyps(EOS-CRSwNP) and further investigate the correlation between IL-13 and MUC5AC in EOS-CRSwNP. METHODS MUC5AC was detected in tissues from normal nasal mucosa and EOS-CRSwNP by immunohistochemistry and ELISA. We also used ELISA to detect the expression of IL-13 in normal nasal mucosa and EOS-CRSwNP. Correlation between IL-13 and MUC5AC in EOS-CRSwNP was investigated by correlation analysis. Secretion of MUC5AC in IL-13 incubated primary air-liquid interface(ALI)-cultured nasal polyp epithelial cells was examined by ELISA. RESULTS MUC5AC is mainly expressed in the epithelium of nasal mucosa by immunohistochemistry. By ELISA, expressions of MUC5AC and IL-13 are higher in EOS-CRSwNP than those in normal nasal mucosa. Correlation analysis shows that there exists a high correlation between MUC5AC and IL-13 in EOS-CRSwNP. IL-13 upregulates expression of MUC5AC in IL-13 incubated primary ALI-cultured nasal polyp epithelial cells for 7 or 14 days. CONCLUSION Expressions of MUC5AC and IL-13 are higher in EOS-CRSwNP than those in normal nasal mucosa respectively, and MUC5AC correlates with IL-13 highly in EOS-CRSwNP.

PURPOSE: Group 2 innate lymphoid cells (ILC2s) are a novel population of lineage-negative cells that induce innate type 2 responses by producing the critical Th2-type cytokines IL-5 and IL-13 in response to IL-25 and IL-33 stimulation. ILC2s accumulation in the peripheral blood of patients with allergic rhinitis (AR) is controversial; the precise role of ILC2s in the immunopathogenesis of AR is still not clear. We investigated the role of ILC2s in phenotypic AR sensitized to distinct allergens. METHODS: Flow cytometric analysis of the peripheral blood of 7 healthy controls (HCs), 9 patients monosensitized to house dust mite (HDM), and 8 patients monosensitized to mugwort was performed to quantify ILC2s frequency. Peripheral blood mononuclear cells (PBMCs) were isolated from HDM-AR and mugwort-AR patients, and Lineage- and Lineage+ cells were separated using a fluorescence-activated cell sorter (FACS). IL-5 and IL-13 levels in the supernatants of PBMCs, and Lineage- and Lineage+ cells stimulated with IL-25 and/or IL-33 combined with IL-2 in vitro were assessed using the Milliplex magnetic bead kit. RESULTS: The percentage of ILC2s was significantly elevated in HDM-AR patients compared to mugwort-AR patients and HCs, while no significant difference was found between mugwort-AR patients and HCs. IL-33+/-IL-25 plus IL-2 induced a significantly greater release of IL-5 and IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. IL-25 plus IL-2 also induced a significantly greater release of IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. Stimulation with IL-33 and/or IL-25 combined with IL-2 also induced a significantly greater IL-5 and IL-13 release from Lineage- cells compared to Lineage+ cells. CONCLUSIONS: AR patients sensitized to HDM or mugwort allergen have distinct phenotypic and functional profiles in ILC2s frequencies. ILC2s mediate major type 2 immunity in the development of HDM-AR and may be a potential therapeutic target.
Allergens ; Artemisia ; Cytokines ; Humans ; Interleukin-13 ; Interleukin-2 ; Interleukin-5 ; Lymphocytes ; Pyroglyphidae ; Rhinitis*

PURPOSE: Chronic rhinosinusitis with nasal polyps (CRSwNP), a mainly Th2 cytokine-mediated disease, often involves mucus secretion. Recent evidence suggests that transmembrane protein 16A (TMEM16A), a calcium-activated Cl- channel (CaCC), can regulate mucus secretion from airway epithelium by transepithelial electrolyte transport and hydration. However, the role of TMEM16A in mucin production/secretion in the airway epithelium is not clear. This study was conducted to determine the role of TMEM16A in mediating mucin secretion in human nasal polyp epithelial cells (HNPECs) induced by IL-13. METHODS: Human sinonasal mucosa tissue and dissociated sinonasal epithelium from control subjects and patients with CRSwNP were assessed for the expression of TMEM16A and the secretion of human mucin 5AC (MUC5AC) by immunohistochemistry, Western blot analysis, and enzyme-linked immuno-sorbent assay (ELISA). A model of the Th2 inflammatory environment was created by exposure of primary air-liquid interface (ALI)-cultured HNPECs to interleukin-13 (IL-13) for 14 days, with subsequent assessment of TMEM16A expression in cell lysates by Western blotting and MUC5AC secretion in apical washings of cells by ELISA. RESULTS: The expressions of TMEM16A and MUC5AC were increased in human nasal polyp tissue and dissociated nasal polyp epithelium. TMEM16A was detected in IL-13-treated HNPECs, specifically in MUC5AC-positive cells but not in ciliated cells. IL-13 treatment increased percentages of TMEM16A-positive cells, MUC5AC-positive cells, and cells coexpressing TMEM16A/MUC5AC, the expression of TMEM16A protein, and the secretion of MUC5AC. T16Ainh-A01, a TMEM16A inhibitor, attenuated these IL-13-induced effects. CONCLUSIONS: The expression of TMEM16A and MUC5AC are increased in CRSwNP, which might be a direct effect of Th2 cytokines present in the sinonasal mucosa in CRSwNP. Down-regulation of TMEM16A expression and MUC5AC secretion in HNPECs by T16Ainh-A01 indicates that TMEM16A might play an important role in mucin secretion in upper airway inflammatory diseases.
Blotting, Western ; Cytokines ; Down-Regulation ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells* ; Epithelium ; Humans ; Immunohistochemistry ; Interleukin-13 ; Mucin 5AC ; Mucins* ; Mucous Membrane ; Mucus ; Nasal Polyps ; Negotiating

Objective To investigated the effects of IL -18 on middle ear allergic inflammation in rat model of OME .Methods Eighteen SD rats were randomly divided into three group :group A(control group ,n=12 ears) , group B (OME model group ,n=12 ears) ,group C (IL -18 injectiion group ,n=12 ears) .The rat model of OME was established by sensitizing with ovalbumin (OVA) and later challenging in tympanic bullae .Recombinant rat IL-18 (1 μg ,+0 .2 ml saline ,) were injected in group C at 1 ,2 ,7 ,8 ,15 ,16 day .At the same time sacle ,0 .2 ml sa-line ,instead of IL -18 ,were intraperitoneal injection in group A and B .The morphologic changes of the middle ear epithelial cells and inflammatory cells infiltration were observed under light microscope .The level of IFN -γand IL-4 in tympanic lavage fluid(TLF) were determined by ELISA .Results Pathological examination showed that middle ear mucosa inflammation and eosinophil infiltration in group C were no less severe than group B .The numbers of neutrophils in group C increased significantly compraring with group B (P<0 .05) .Numbers of eosinophils in group C were slightly increased comparing with group B (P>0 .05) ,while significantly greater than that in group A (P<0 .05) .ELISA showed that the level of IFN -γ in group C was stronger than that in group B and A (both P<0 .05) .As compared to the group A ,the expression of IL -4 in group B and group C were remarkably stronger (both P<0 .05) ,no significant difference was found between group B and group C (P>0 .05) .Addtionally ,there was no significant difference in the ratio of Th1 /Th2(IFN -γ/IL -4)between group B and group C (P>0 .05) . Conclusion IL -18 acts as an immune regulatory factors ,significantly increases Th1 cytokine IFN -γ.Although to some extent alleviate the OME rat middle ear Th1 /Th2 imbalance ,there is still excessive activation of Th cells . Th1 and Th2 cells factor are excessive for the secretion disorder of the immune response status .The OME rat mid-dle ear allergic inflammation has not been fundamentally alleviated ,the underlying mechamism should be further studied .

Objective To analyze the clinical feature and treatment methods of Artemisia pollinosis.Methods Skin prick test results of 14 426 cases from Beijing Tongren hospital and pollen concentration of Beijing observatory from 2007 to 2011 were analyzed to identify the clinical feature of Artemisia pollinosis patients and its correlation with the pollen concentration.Patients were given leukotriene receptor antagonists (Montelukast) for 2 weeks,followed by 4 weeks of mometasone furoate nasal spray (EIT group:n =21),or only 4 weeks of mometasone furoate nasal spray (POT group:n =16).The nasal symptom score was compared between 2 groups.SPSS 16.0 software was used to analyze the data.Results Artemisia pollinosis accounted for 30.8％ (4 442/14 426) of all SPT positive allergic rhinitis patients,and most Artemisia SPT positive results were strong positive (3 793/4 442,85.4％) ; onset age peak of Artemisia pollinosis patients was at the age of 19 to 30,onset time concentrated in August to September,was consistent with the peak period of Artemisia pollen concentration; EIT treatment using leukotriene receptor antagonists two weeks before pollen season significantly improved sneeze,sniveling and rhinocnesmus symptoms (t value was 3.28,3.92,3.09,respectively,all P ＜ 0.01) compared with post-onset treatment(POT).But nasal obstruction and cough symptoms had no significant difference between two groups (t value was 0.85,1.52,respectively,all P ＞ 0.05).Conclusion Artemisia pollen is the main pollen allergen in Beijing,EIT treatment was effective to pollinosis.

OBJECTIVE: To determine the inflammatory cells in the mucosa at the medial aspect of the normal uncinate process compared with that on the protected lateral aspect of the normal uncinate process. METHOD: The mucosa of 20 uncinate process from the nasal cavity of 17 patients with no evidence of sinus disease undergoing functional endoscopic sinus surgery were recruited for the study. The material was stained with HE, Chromotrope 2R, Alcian blue-periodic acid-schiff, Toluidine blue. Specimens were observed using an Olympus microscope. RESULT: The number of mast cells and goblet cells were found to be higher on the lateral aspect of the normal uncinate process than on the medial aspect. The number of plasma cells was obviously different from that of lymphocytes. We did not found any eosinophils on either sides of uncinate process. CONCLUSION There are differences in the number of mast cells and goblet cells between the mucosa at the medial aspect of the normal uncinate process and the mucosa at the protected lateral aspect of the normal uncinate process.
Adolescent ; Adult ; Ethmoid Sinus ; pathology ; Female ; Goblet Cells ; cytology ; pathology ; Humans ; Male ; Mast Cells ; cytology ; pathology ; Middle Aged ; Nasal Mucosa ; pathology ; Paranasal Sinuses ; pathology ; Young Adult

OBJECTIVE: To investigate the electrophysiological characteristics and pathological changes in different recurrent laryngeal nerve injury models. METHOD: Twenty dogs were divided into different models of recurrent laryngeal nerve injury. Electrophysiological changes of animal models and histopathological changes in laryngeal muscles were investigated at different time. RESULT: Amplitude of MUP in TA decreased at first and then increased a little, while duration of MUP keeps decreased. The latent periods of the evoked potential in the incomplete injury group increased at first and then went down. In animal histopathological investigation, the lighter of the muscle injuries and the bigger of the diameter of muscle fibers and fascicles, and the smaller of the number of cellular nucleolus per square inch. In the early periods after injury, the correlation among the standard amplitude, threshold amplitude, maximum amplitude of EP and the diameter of fascicles were positive; while in the late period, the correlation among the amplitude, duration, area of MUP and nucleolus number was positive. CONCLUSION The physiological characteristics of laryngeal nerves and muscles could be known. LEMG was an important method in diagnosis of laryngeal nerve and muscle diseases.
Animals ; Disease Models, Animal ; Dogs ; Electromyography ; Evoked Potentials ; Laryngeal Muscles ; innervation ; physiopathology ; Recurrent Laryngeal Nerve Injuries

OBJECTIVE: To investigate the mechanism of hearing pathway NMDAR regulating the changes of phosphorylated c-Jun expression in spiral ganglion after exposed Kunming mice to neural injury noise stimulation to induce permanent or temporary threshold shift. METHOD: To compare the different expressions of the key components (phosphorylated C-Jun) of NMDAR signal pathway during neural injury stimulation by Immunohistochemistry in CG. RESULT: The levels of phosphorylated c-Jun remarkably, increased in the spiral ganglion after 8 h, 48 h, 7 d and 14 d following noise trauma induced permanent threshold shift (PTS), and the numbers of positive cells reduced gradually. The similar changes occur in mice treated with MK-801 30 minutes before and after 3 h trauma induced PTS. After 48 h of noise induced TTS, the expression of Phosphorylated c-Jun return the level of normal control. CONCLUSION The expressions of phosphorylated c-Jun are time-related and uniform in the time and position in CG after noise trauma. MK-801 can alleviate the damage of noise trauma by altered the NMDA receptor-mediated calcium influx. Therefore, the NMDA receptors may involved in the damage of inner ear in common.
Acoustic Stimulation ; Animals ; Auditory Threshold ; Cochlea ; metabolism ; Hearing Loss, Noise-Induced ; metabolism ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Mice ; Mice, Inbred Strains ; Spiral Ganglion ; metabolism

OBJECTIVE: To study the changes of N-methyl-D-aspartate (NMDA) receptor subunit 2A (NR2A) expression at local synapses in auditory cortices after early postnatal sound insulation and tone exposure. METHOD: We prepared highly purified synaptosomes from primary auditory cortex by Optiprep flotation gradient centrifugations, and compared the differences of NR2A expression in sound insulation PND14, PND28, PND42 and Tone exposure after sound insulation for 7 days by Western blotting. RESULT: The results showed that the NR2A protein expression of PND14 and PND28 decreased significantly (P<0.05). Tone exposure after sound insulation for 7 days, mSIe NR2A protein level increased significantly (P<0.05). It showed bidirectional regulation of NR2A protein. No significant effects of sound insulation and lone exposure were found on the relative expression level of NR2A of PND42 (P>0.05). CONCLUSION The results indicate that sound insulation and experience can modify the protein expression level of NR2A during the critical period of rat postnatal development. These findings provide important data for the study on the mechanisms of the developmental plasticity of sensory functions.
Acoustic Stimulation ; Animals ; Auditory Cortex ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Synapses ; metabolism

OBJECTIVE: To investigate the T helper cells (Th) predominant differentiation and the modulation of nuclear transcription factors kappa B (NF-kappaB) in middle ear of rat model of otitis media with effusion (OME). METHOD: Sixteen SD rats were randomly divided into OME (Exp group) and control group (Con group). The expression of NF-kappaB were observed by immunohistochemistry. The level of IFN-gamma and IL-4 in tympanic lavage fluid (TLF) were determined by ELISA. RESULT: As compared to the Con group , the level of IL-4 and the ratio of Th2/Th1 (IL-4/IFN-gamma) in TLF of Exp group significantly increased (P<0.05), when no significant difference in IFN-gamma levels in TLF was found. The ratio of NF-kappaB p65 positive cells to white cells in temporal bone marrow smears and middle ear mucosa of Exp group was significantly higher than that of Con group (P<0.05). The expression of NF-kappaB p65 in temporal bone marrow smears and middle ear mucosa was signficantly positively correlated with the concentration of IL-4 in TLF of Exp group (P<0.05). CONCLUSION The middle ear is capable of mounting an allergic response and subsequent formation of effusion. There is Thl/Th2 immune response imbalance, which polarizes toward Th2 response in the middle ear microenvironment of allergic OME rat model. Moreover , NF-kappaB may participate in regulating Th2 predominant reaction.
Animals ; Interleukin-4 ; metabolism ; Male ; NF-kappa B ; metabolism ; Otitis Media with Effusion ; chemically induced ; immunology ; metabolism ; Ovalbumin ; adverse effects ; Rats ; Rats, Sprague-Dawley ; Th1 Cells ; immunology ; Th2 Cells ; immunology