Effects and clinical significance of NLRP3 inflammasome activated by IL-17A in CRSwNP / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the effects and clinical significance of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activated by interleukin (IL)-17A in chronic rhinosinusitis with nasal polyps (CRSwNP). Methods: Patients underwent nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-sen University from January 2020 to December 2021 were collected, including 28 CRSwNP (including 19 males and 9 females, aged 19 to 67 years), 22 chronic rhinosinusitis without nasal polyps (CRSsNP) and 22 controls. qRT-PCR was used to detect the expressions of IL-17A, NLRP3, IL-1β and IL-18 in the three groups, and their correlations were analyzed. The positions of IL-17A, NLRP3 and IL-18 in nasal polys were analyzed by immunofluorescence. Western Blotting and ELISA were employed to detect the expression of NLRP3, IL-1β and IL-18 in the human nasal epithelial cells after using IL-17A stimulation or IL-17A receptor inhibitor. Immunofluorescence was used to observe the NLRP3, IL-1β, and IL-18 protein expression after IL-17A stimulating human nasal epithelial cells, and after the use of IL-17A receptor inhibitor and NLRP3 inhibitor MCC950. The correlations between NLRP3, IL-1β, IL-18 and CT scores, nasal endoscopic scores, visual analogue scale (VAS) scores, and sino-nasal outcome test (SNOT) 22 scores of CRSwNP patients were analyzed. SPSS 20.0 software was used for statistical analysis. Results: The expressions of IL-17A, NLRP3, IL-1β and IL-18 in the tissues of CRSwNP patients were significantly higher than those in CRSsNP group(P=0.018,P<0.001,P=0.005, P=0.016) and the control group(all P<0.001). IL-17A was positively correlated with the expression of NLRP3, IL-1β, and IL-18(r ralue was 0.643,0.650,0.629,respectively, all P<0.05). IL-17A, NLRP3, and IL-18 were co-localized in the epithelial propria of polyp tissue. IL-17A stimulated the expressions of NLRP3, IL-1β, and IL-18 in human nasal epithelial cells. After the use of IL-17A receptor inhibitor, the expressions of NLRP3, IL-1β, and IL-18 were significantly down-regulated. After the use of NLRP3 inhibitor MCC950, IL-17A was significantly down-regulated to promote the expression of NLRP3, IL-1β, and IL-18. The expressions of NLRP3, IL-1β and IL-18 were positively correlated with CT, nasal endoscopy, VAS, and SNOT22 scores in patients with CRSwNP. Conclusions: IL-17A promotes the release of IL-1β and IL-18 by activating the NLRP3 inflammasome and aggravates the severity of the disease in CRSwNP.

Expression of CD8+Treg cells in chronic rhinosinusitis and its correlation with eosinophilic infiltration / 中华耳鼻咽喉头颈外科杂志

Objective: To detect the percentages of CD8+Treg cells in the nasal mucosa and peripheral blood of chronic rhinosinusitis (CRS) and to explore their correlation with eosinophilic infiltration. Methods: Thirty-three chronic rhinosinusitis with polyp (CRSwNP), 26 chronic rhinosinusitis without polyp (CRSsNP) and 27 control patients who were collected with the nose mucosal tissue and peripheral blood in the Third Affiliated Hospital of Sun Yat-sen University from March 2017 to October 2018 were selected, including 59 males and 27 females, aging from 18 to 72 years. Hematoxylin and eosin (HE) staining was used to observe the number of eosinophils in the nasal tissues and to classify the CRS into eosinophilic CRS (ECRS) and non-eosinophilic CRS (Non-ECRS). Flow cytometry was used to detect the percentages of CD4+ and CD8+T cells in lymphocytes of nasal mucosa and peripheral blood. The percentages of CD8+Foxp3+Treg cells, CD8+Foxp3-IL-10+Treg cells, CD8+IFN-γ+T cells (Tc1), CD8+IL-4+T cells (Tc2) and CD8+IL-17A+T cells (Tc17) in lymphocytes of nasal mucosa and peripheral blood were also tested. Besides, the percentages of Foxp3+TGF-β+Treg cells and Foxp3+IL-10+Treg cells in CD8+T cells were determined. All data were represented by M (IQR). GraphPad 7.0 and SPSS 16.0 were used for illustration and statistical analysis. Results: The percentage of CD8+T cells (37.75%(17.35%)) was higher than that of CD4+T cells (4.72%(4.29%)) in nasal mucosa (Z=-5.70, P<0.001), while lower (23.60%(9.33%)) than that of CD4+T cells (44.05% (10.93%)) in peripheral blood (t=9.72, P<0.001). CRSwNP patients possessed the highest Tc2 (1.82% (1.22%)) and Tc17 (1.93% (2.32%)) percentages than CRSsNP (Tc2: 0.84% (0.79%); Tc17: 0.54% (1.04%)) and control (Tc2: 1.09% (0.92%); Tc17: 0.47% (0.51%), both P<0.05) patients. While, CRSwNP patients possessed the lowest CD8+Foxp3+Treg cells percentage (0.10% (0.32%)) than CRSsNP (0.43% (1.45%)) and control (0.48% (0.83%), Z value was -2.24, -2.22, respectively, P value was 0.025, 0.027, respectively). The percentages of Foxp3+TGF-β+Treg cells and Foxp3+IL-10+Treg cells of CD8+T cells in nasal mucosa in CRSwNP were also lower than controls (Z value was 1.46, 0.49, respectively, both P=0.001). Moreover, the percentage of CD8+Foxp3-IL-10+Treg cells of CD8+T cells was decreased in nasal mucosa of CRSwNP patients (0.14% (0.28%)) when compared with that of CRSsNP (0.89% (0.81%), Z=0.61, P=0.03). ECRS patients had the lower percentages of CD8+Foxp3+Treg cells (0.07% (0.44%)) and CD8+Foxp3-IL-10+Treg cells (0.13% (0.21%)) than Non-ECRS patients (CD8+Foxp3+Treg cells: 0.53% (0.75%); CD8+Foxp3-IL-10+Treg cells: 0.29% (0.76%), t value was 2.14, 2.78, respectively, both P<0.05). The percentage of CD8+Foxp3+Treg cells and the ratio of CD8+Foxp3-IL-10+T per CD8+T cells were negatively correlated with the percentage of eosinophils in CRS patients(R2 value was 0.56, 0.78, respectively, both P<0.001). There was no significant difference in the distribution of CD8+Fxop3+Treg cells and CD8+Fxop3-IL-10+Treg cells in peripheral blood among different groups. Conclusion: The percentages of CD8+Treg cells decrease in CRSwNP patients, especially in ECRS patients, which are opposite to that of Tc2 and Tc17, and negatively correlate with the eosinophils percentage. This indicates that the decrease in the ratio of CD8+Treg cell may be associated with the immune-imbalance and eosinophilic infiltration in nasal mucosa of CRS patients.

The expression of Notch pathway in nasal polyps and its correlation with the decreased Treg percentage and eosinophil infiltration / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the correlation between Notch pathway expression in nasal polyps and Treg percentage and Eos infiltration. Methods: Patients with chronic sinusitis and simple nasal septum deviation who received nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-Sen University between November 2012 and August 2018 were selected and enrolled in CRS group and control group respectively. Nasal mucosa tissues were collected from 30 CRSsNP patients (14 males and 16 females aged from 18 to 63), 58 CRSwNP patients (38 males and 20 females aged from 18 to 65) and 29 patients (19 males and 10 females aged from 20 to 57), who underwent nasal endoscopic surgery for correction of simple nasal septum deviation. Hematoxylin-eosin(HE) staining was used to observe the infiltration of eosinophilic granulocytes in the tissues and to classify chronic sinusitis with polyps (CRSwNP) into eosinophilic chronic rhinosinusitis with nasal polyps (Eos-CRSwNP)and non-eosinophilic chronic rhinosinusitis with nasal polyps (Eos-CRSwNP). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of Notch pathway receptors (Notch-l, 2, 3, 4) and their ligands (Jagded-l, Jagded-2, Delta-l, Delta-3and Delta-4) in the nasal mucosa of each group, as well as the expression of Th2 cytokines (IL-4, IL-5, IL-13), eosinophilic cationic protein (ECP)and the key transcription factor Foxp3 in Treg cells. Finally, flow cytometry was used to detect CD4+CD25+Foxp3+ Treg cells in nasal mucosa of each group. Results: Compared with controls, the expression of Th2 cytokines (IL-4, IL-5, IL-13) in CRSsNP and non-Eos-CRSwNP patients was the highest in Eos-CRSwNP (F=16.930,9.197,9.116, all P<0.05). Foxp3 had the lowest expression in Eos-CRSwNP patients and was lower than non-Eos-CRSwNP patients (F=2.780,P<0.05), and was negatively correlated with ECP (r=-0.326,P<0.05). Compared with controls, Eos-CRSwNP patients in CRSsNP patients and non-Eos-CRSwNP patients exhibited a significantly lower frequency of CD4+CD25+Foxp3+Treg cells (F=13.140, all P<0.01). The expression of Notch-l and Jagged-l in Eos-CRSwNP was significantly higher than that of the controls, CRSsNP patients and non-Eos-CRSwNP patients (F=5.953/F=6.380, P<0.05). In the nasal polyp group, the expression of Notch-l and Jagged-l showed significantly negative correlation with Foxp3 (r=-0.611/-0.346, all P<0.05), and positive correlation with Th2 cytokines (IL-4, IL-5, IL-13) and ECP, respectively (r=0.781/0.459,0.621/0.601,0.605/0.490,0.464/0.668, all P<0.05). There was no significant difference in the expression of receptor and ligand of the other Notch pathway among the groups. Conclusion: Abnormal activation of Notch-l/Jagged-l pathway may be involved in decreasing Treg ratio in Eos-CRSwNP, thereby promoting Th2 inflammatory response and Eosinophil infiltration.

miR-18a enhances the radiosensitivity of nasopharyngeal carcinoma cells through inducing autophagy / 中华耳鼻咽喉头颈外科杂志

Objective: To explore the impacts of miR-18a overexpression or depression on the radiosensitivities of nasopharyngeal carcinoma cell line CNE1 and CNE2 and underlying mechanisms. Methods: CNE1 and CNE2 were transfected with miR-18a mimics, inhibitor and the corresponding control vectors. qRT-PCR and western blot were used to determine the ataxia telangiectasia mutated (ATM) expressions in CNE1 and CNE2. CNE1 and CNE2 with stably expressing miR-18a and miR-18a siRNA were constructed. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the impacts of the miR-18a overexpression or depression combined with irradiation on the cell growth. Flow cytometry was used to detect the cell apoptosis and cell cycle. Colony formation assay was used to evaluate the raodiosensitivities of cells. Acridine orange (AO) staining and western blot were used respectively to test the autophagy and the expressions of related proteins. Independent samples t test was used to compare the mean value between groups by using SPSS 16.0. Results: ATM mRNA was decreased significantly in CNE1 and CNE2 cells transfected with 100 or 200 nmol/L miR-18a mimics for 48 hours (CNE1: RQ=0.174±0.139 and 0.003±0.001, t=9.939 and 19 470.783;CNE2: RQ=0.024±0.008 and 0.019±0.012, t=270.230 and 137.746, respectively, all P<0.001). ATM proteins were also decreased after transfected with 100 or 200 nmol/L miR-18a mimics for 72 hours. While in the cells transfected with 100 and 200 nmol/L miR-18a inhibitor for 48 hours, the expressions of ATM mRNA were upregulated significantly (CNE1: RQ=9.419±2.495 and 2.500±1.063, t=-4.427 and -41.241; CNE2: RQ=7.210±0.171 and 115.875±15.805, t=-62.789 and -12.589, all P<0.05), and the expressions of ATM proteins increased after transfected for 72 hours. The growth of cells with miR-18a overexpression plus 4 Gy irradiation were obviously inhibited compared to that of cells with the 4Gy irradiation alone; while the growth of miR-18a-inhibited cells increased compared to that of cells with 4 Gy irradiation alone (all P<0.05). CNE1 transfected with 100 nmol/L miR-18a mimics plus 4 Gy irradiation showed the higher apoptosis rate than the cells with 4 Gy irradiation alone ((22.9±2.1)% vs. (16.3±1.0)%, t=-4.870, P<0.01). Compared to the cells with 4 Gy irradiation alone, miR-18a-overexpressed cells plus 4 Gy irradiation decreased their percentages in G1 phases ((20.2±3.0)% vs. (29.8±4.4)%, t=3.119) and G2/M phases ((21.5±0.9)% vs. (33.4±3.1)%, t=6.410, P<0.05 for both), and increased their percentages in S phases ((56.7±4.9)% vs. (36.8±6.4)%, t=-4.246, P<0.05), and these cells possessed less colony number after exposure to different doses of irradiation, more autophagy-lysosome number, and more expressions of LC3 proteins (all P<0.05). There were no significant differences in the expressions of p62 expressions between different groups of cells. Conclusion: Overexpression of miR-18a can enhance the radiosensitivities of NPC cells by targeting ATM to abrogate G1/S, G2/M arrest and to induce autophagy and apoptosis.

Construction of Human Nasophayngeal Cancer Cells Stablely Expressing miR-18a and miR-18a siRNA / 中山大学学报(医学科学版)

[Objective]To construct miR-18a overexpression and inhibition lentivirus vectors and to determine their effects on human nasopharyngeal cancer(NPC)cell line CNE1 and CNE2.[Methods]Designed the primers for Real-time polymerase chain(PCR)reaction to obtain the miR-18a premature gene.The premature gene and the siRNA oligo-nucleutides of miR-18a were connected to the lentivirus vector GV369 and GV280,respectively.The construction vectors were confirmed by DNA sequencing.Then,293T cell was infected with the vectors plus Helper 1.0 and pHelper 2.0 vec-tors to obtain recombinant lentivirus vector for miR-18a overexpression and inhibition. The NPC cell line CNE1 and CNE2 were infected with the successful recombinant lentivirus vectors.Puromycin was added to select the positive infect-ed cells. PCR method was used to detect the miR-18a expression level after infecting the recombinant lentivirus vector into the NPC cell line.[Results]A recombinant lentivirus vector expressing miR-18a interference oligonucleutides was obtained and confirmed by DNA sequencing.The virus titer was 3×108TU/mL,and the expression of its target gene ATM was downregulated in CNE1 and CNE2.A recombinant lentivirus vector expressing miR-18a premature gene was obtained and confirmed by DNA sequencing. The virus titer was 3×108TU/mL,and the miR-18a was overexpressed in CNE1 (20.3 fold upregulation,P<0.01)and CNE2(122.5 fold upregulation,P<0.01),and its target gene ATM was downregu-lated.[Conclusions]The miR-18a overexpression and suppression lentivirus vectors are successfully constructed.These vec-tors could alter the expression level of miR-18a in NPC cell line significantly,and provide a stable cell line for functional studies in the future.

Vγ1+γδT Cells Are Correlated With Increasing Expression of Eosinophil Cationic Protein and Metalloproteinase-7 in Chronic Rhinosinusitis With Nasal Polyps Inducing the Formation of Edema

PURPOSE: We have found that expression of γδT cells is increased in pathological mucosa of chronic rhinosinusitis with nasal polyps (CRSwNP) compared with normal nasal mucosa. This increase is correlated with the infiltration of eosinophils in CRSwNP. Here, we investigated the expression of γδT cells, inflammation and tissue remodeling factors as well as their probable relationships in different types of chronic rhinosinusitis (CRS) in China. METHODS: A total of 76 surgical tissue samples that included 43 CRSwNP samples (15 eosinophilic and 28 non-eosinophilic), 17 CRS samples without nasal polyps (CRSsNP), and 16 controls were obtained. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the mRNA expression levels of Vγ1⁺γδT cells, Vγ4⁺γδT cells, eosinophil cationic protein (ECP), interleukin (IL)-8, transforming growth factor (TGF)-β2, metalloproteinase (MMP)-7, tissue inhibitor of metalloproteinase (TIMP)-4 and hypoxia-inducible factor (HIF)-1α. Enzyme linked immunosorbent assay (ELISA) was used to measure the protein level of ECP and MMP-7 in CRSwNP. The eosinophils were counted and the level of edema was analyzed with HE staining. RESULTS: The mRNA expression levels of the Vγ1 subset, ECP and MMP-7 were significantly increased in CRSwNP with histological characteristics of eosinophilic infiltration and edema. The expression of the Vγ1 gene in CRSwNP correlated positively with the expression of both ECP and MMP-7. No significant decreases in the mRNA expression levels of TGF-β2, TIMP-4 or HIF-1α were observed in the CRSwNP samples. The expression levels of Vγ1 gene, ECP and MMP-7 were significantly increased in eosinophilic CRSwNP compared to non-eosinophilic CRSwNP. CONCLUSIONS: Our results suggest the associations between Vγ1⁺γδT cells, ECP and MMP-7 in CRSwNP, indicating that Vγ1⁺γδT cells can induce the eosinophilic inflammation, which has a further effect on the formation of edema.

Expression of peripheral blood gammadelta T cells, treg cells and cytokines IL-17 and TGF-beta1 in patients with allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of peripheral blood gammadelta T cells/CD4 CD25+ regulatory T cells(Treg) and cytokines interleukin 17 (IL-17) and transforming growth factor beta1 (TGF-beta1) in patients with allergic rhinitis.</p><p><b>METHODS</b>From March 2012 to July 2012, 32 patients with allergic rhinitis (AR group) and 20 healthy control subjects (control group) were collected. The expression of peripheral blood gammadelta T cells/Treg cells were measured by flow cytometry and the levels of IL-17 and TGF-beta1 were evaluated by ELISA. SPSS 16.0 software was used to analyze the data.</p><p><b>RESULTS</b>The percentages of gammadeltaT cells in AR group were (13.30 +/- 8.62)%, which was significantly higher (t = 5.18, P < 0.01) than those in control group (5.18 +/- 1.86)%. The percentages of Treg cells in AR group were (1.75 +/- 0.56)%, which were significantly lower (t = 7.46, P < 0. 01) than those in control group (4.76 +/- 1.74)%. The IL-17 levels in AR group were (668.55 +/- 45.15) pg/ml, which were also significantly higher (t = 8.97, P < 0.01) than those in control group (573.53 +/- 17.42) pg/ml. The TGF-beta1 levels in AR group were (0.34 +/- 0.04) pg/ml, which were also significantly lower (t = 9.51, P < 0.01) than those in control group (0.49 +/- 0.06) pg/ml. There was a negative correlation between the percentages of gammadelta T cells and Treg cells (r = -0.561, P < 0.01). There was a negative correlation between the percentages of gammadelta T cells and TGF-beta1 levels (r = -0.622, P < 0.01). A positive correlation was shown between the percentages of gammadelta T cells and IL-17 levels in AR (r = 0.469, P < 0.01). A positive correlation was shown between the percentages of Treg cells and TGF-beta1 levels in AR (r = 0.738, P < 0.01). There was no correlation between IL-17 levels and the percentages of Treg cells or TGF-beta1 levels (r value was -0.111, -0.196, all P > 0.05).</p><p><b>CONCLUSION</b>There are imbalances of gammadelta T and Treg cells in peripheral blood of patients with allergic rhinitis. gammadelta T cells may be the main cell to produce IL-17, which may play an important role in allergic rhinitis.</p>

Radiobiological characteristics and MRN complex expression of human nasopharyngeal carcinoma cell lines / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the radiobiological characteristic of human nasopharyngeal carcinoma cell lines CNE1 and CNE2 and the changes in expression MRN (Mre11-Rad50-Nbs1) complex in the cell lines exposed to irradiation.</p><p><b>METHODS</b>CNE1 and CNE2 were irradiated by a linear accelerator. Radiobiological characteristics were detected by colony assay and MTT assay. MRN complex expression were examined by Western blot.</p><p><b>RESULTS</b>Surviving fraction at 2 Gy (SF2), quasi-threshold Dose (Dq), and mean lethal dose (Do) of CNE1 were 0.56, 1.449 Gy and 1.480 Gy; SF2, Dq, and Do of CNE2 were 0.44, 0.776 Gy and 1.685 Gy, respectively. Survival fraction of CNE1 at the day 6 after 4 Gy irradiation was 0.59 and that of CNE2 was 0.79 when compared with control, with the up-regulated expressions of Rad50 in CNE1 and Mre11, Rad50 and Nbs1 in CNE2 (P < 0.05).</p><p><b>CONCLUSIONS</b>CNE1 and CNE2 were sensitive to radiation, but there were radioresistance cells in CNE2. The expressions of some components of MRN complex were up-regulated to repair DNA lesions induced by radiation.</p>

γδ T cell expression and significance in chronic rhinosinusitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the expression of γδ T cells in chronic rhinosinusitis (CRS) and its potential significance in pathogenesis.</p><p><b>METHODS</b>γδ T cell expression was detected by immunohistochemistry (Envision method). From polyps (25 CRS patients with nasal polyps, CRSwNP), inferior turbinate mucosa (13 CRS patients without nasal polyps, CRSsNP), and 16 inferior turbinate mucosa from patients with deviation of nasal septum served as control. The infiltration of eosinophils in eosinophilic CRSwNP was observed by HE staining. The differences of expression of γδ T cells between each groups were compared, meanwhile the relationship between γδ T cells and eosinophils were analyzed. SPSS 16.0 software was used to analyze the data.</p><p><b>RESULTS</b>The positive range of γδ T cells in CRSwNP group and CRSsNP group was 88.0% and 84.6%, respectively, both higher than 37.5% in control group (χ(2) = 13.413, P < 0.01, χ(2) = 6.564, P < 0.05, respectively), CRSwNP group had no statistical significance compared with CRSsNP group (χ(2) = 0.086, P > 0.05). The expression of γδ T cells in CRSwNP group was stronger than CRSsNP group and control group (U = 596, P < 0.01, U = 296, P < 0.01, respectively); CRSsNP group was stronger than control group (U = 216, P < 0.05). There was a positive correlation between γδ T cells and eosinophils (r = 0.579, P < 0.05).</p><p><b>CONCLUSIONS</b>The expression of γδ T cells was increased in nasal mucosa of CRS. γδ T cells may be involved in the pathogenesis of CRS.</p>

Distribution and clonality of T cell receptor Vγ and Vδ subfamily in peripheral blood of patients with allergic rhinitis before and after immunotherapy / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the distribution and clonality of T cell receptor (TCR) Vγ and Vδ subfamily in peripheral blood of patients with allergic rhinitis before and after 1 year treatment with immunotherapy.</p><p><b>METHODS</b>The specific IgE and the complementary determinant region 3 (CDR3) of TCR V γ (I-III) and Vδ(1-8) subfamily genes in mononuclear cells were amplified from 10 effective cases of allergic rhinitis before and after 1 year treatment with immunotherapy, to observe the distribution and utilization of TCR Vγ and Vδ repertoire. The positive PCR products were further labeled with RT-PCR and analyzed by gene scan technique to determine the CDR3 size and evaluate the clonality of the detectable TCR Vγ and Vδ T cells. Peripheral blood of 10 healthy adults served as controls.</p><p><b>RESULTS</b>All symptoms were significantly improved after 1 year specific immunotherapy, but no changes were seen in specific IgE [(22.89 ± 9.60) kU/L before treatment, (19.62 ± 7.63) kU/L after treatment, Z = 1.051, P > 0.05]. No statistically significant differences of expression levels of the TCR Vγ I-III subfamily genes were found between patients with allergic rhinitis normal control group (t value were -0.679, -0.516, -0.808, all P > 0.05), but significantly decreased after 1 year treatment. There were statistically significant differences of expression levels of the TCR VγI-II subfamily genes before and after treatment (t value were -2.904, -2.217, all P < 0.05). 5.30 ± 0.82, 4.90 ± 0.57 and 5.20 ± 1.40 out of TCR Vδ (1-8) subfamilies were selectively expressed in T cells in patients with allergic rhinitis before and after 1 year treatment and normal control group, predominantly for TCR Vδ 1, 2, 3 and 6. The TCR Vδ 6 subfamily was found to have statistically significant differences in these groups (Fisher's Exact Test, P < 0.05). Compared with the normal control group and the allergic rhinitis group before treatment, a significant higher frequency of Vδ 6 oligoclonal was identified in T cells in patients with allergic rhinitis after 1 year treatment.</p><p><b>CONCLUSIONS</b>There was difference in the expression levels of the TCR Vγ I-III subfamily genes and distribution and clonality of TCR Vγ and Vδ subfamily T cells in peripheral blood of patients with allergic rhinitis before and after 1 year treatment. Specific immunotherapy can be effective in alleviation of the symptom in patients with allergic rhinitis during the early stage, possibly by inducing TCR γδ T cells, especially the TCR Vδ6 subfamily, and possibly no significant relativity between symptom and specific IgE.</p>

Synergistic effects of tetrandrine on the antifungal activity of topical ketoconazole cream in the treatment of dermatophytoses: a clinical trial / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the synergistic effects of tetrandrine (TET) on the antifungal activity of topical ketoconazole (KCZ) in the treatment of dermatophytoses.</p><p><b>METHODS</b>The minimum inhibitory concentrations (MICs) for KCZ and combined KCZ and TET were compared in vitro. A randomized, double-blind trial was conducted among 97 patients with dermatophytoses who were assigned to 3 groups and received: treatment with combination of 2% KZC and 2% TET cream (KCZ + TET group), or only 2% KZC cream (KCZ group), or 2% TET cream (TET group). Patients with tinea corporis and/or tinea cruris were treated for 2 weeks, separately. The patients with tinea pedis and/or tinea manuum were treated for 4 weeks.</p><p><b>RESULTS</b>Compared with KZC alone, combined use of KZC and TET showed lower MICs against clinical isolates of dermatophytes (P<0.05 for all). In the patients with tinea corporis and/or tinea cruris, the rates of overall cure (clinical cure plus mycologic clearance) were 81.25% vs. 33.33% for combined treatment and KZC monotherapy, respectively, after 4 weeks. All clinical indices were significantly different between the combination therapy and only KCZ therapy groups (P<0.05). Among the patients with tinea pedis and/or tinea manuum after 4 weeks treatment, the overall cure rates in the KCZ + TET group and KCZ group were 75.00% vs. 40.00%, respectively. In the KCZ + TET group, all the clinical indices were significantly better than those in the KCZ group and TET group (P<0.05). The rates of overall efficacy in the TET group were all zero. No local skin redness or itching was observed during TET treatment. No clinically significant changes were found in post-treatment routine blood, urine, or stool tests, ECG, or tests for liver and kidney function; no serious adverse events occurred.</p><p><b>CONCLUSION</b>TET synergistically enhanced the clinical efficacy of topical KZC cream in the treatment of dermatophytoses.</p>

Endoscopic endonasal anatomy of pterygopalatine fossa and infratemporal fossa: comparison of endoscopic and radiological landmarks / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility and reliability of the measurement of critical anatomic landmarks of endoscopic endonasal anatomy of pterygopalatine fossa and infratemporal fossa using multislice spiral computed tomography (MSCT), and to illustrate the spatial relationship of the surgical landmarks in pterygopalatine fossa and infratemporal fossa through an endoscopic endonasal view and radiological images.</p><p><b>METHODS</b>Included in this study were eleven fixed cadaver heads (22 pterygopalatine fossa and infratemporal fossa), which were prepared from MSCT scans for establishing a spatial coordinates system to calculate the radiological anatomic data and attaining 3D reconstruction image, and also were anatomically dissected to get anatomic data. The anatomic data in two groups were compared, the endoscopic and radiological images of the same regions acquired during the endoscopic endonasal approaches observed.</p><p><b>RESULTS</b>The distance (x(-) ± s) from nasal columella to sphenopalatine foramen, pterygoid canal, foramen rotundum, foramen ovale, foramen spinosum, carotid canal, foramen lacerum in radiological group were: (68.83 ± 3.00), (72.49 ± 2.88), (75.26 ± 3.14), (88.55 ± 5.00), (95.19 ± 4.31), (106.76 ± 3.77), (88.16 ± 2.87) mm and in anatomic group were: (68.90 ± 3.04), (72.73 ± 3.08), (75.44 ± 3.07), (89.75 ± 4.13), (96.22 ± 3.37), (106.68 ± 3.75), (88.47 ± 2.64) mm. There was no statistical difference between two groups (t value were -0.856, -1.134, -0.920, -1.923, -1.903, 2.820 and 1.209, respectively, all P > 0.05). Sphenopalatine foramen, pterygoid canal, foramen rotundum, foramen ovale, foramen spinosum, foramen lacerum, carotid canal were the corresponding anatomic structures in endoscope and radiology, which provided the surgeons with anatomic landmarks to identify the spatial relationship of the surgical structures in pterygopalatine fossa and infratemporal fossa.</p><p><b>CONCLUSIONS</b>MSCT measurements of anatomic landmarks are feasible and reliable, can be used in clinical individualized surgery. The corresponding anatomic structures of endoscopic and radiological landmarks provide useful reference to surgeons when operating in these areas through an endoscopic endonasal approach.</p>

Regulation of NF-κB signal transduction pathway on cytokines in cultured nasal epithelium / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the activation and regulation of nuclear factor-kappa B (NF-κB) on transcription of cytokines in cultured lipopolysaccharide (LPS)-induced nasal epithelial cells.</p><p><b>METHODS</b>Normal sphenoid mucosa epithelium from 11 patients who accepted pituitary tumor surgery via trans-sphenoid approach was separated and cultured without serum. The epithelium of the third or the forth passage was induced with LPS. Wedelolactone, a blocking agent of NF-κB was used at the same time. An electrophoretic mobility shift assay was used to detect DNA-binding activity of NF-κB. The reverse transcription-polymerase chain reaction was used to evaluate mRNA of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-5, IL-6, IL-8, granulocyte-macrophage colony stimulating factor (GM-CSF), regulated on activation, normal T expressed and secreted (RANTES), eotaxin, eotaxin-2, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Statistical analysis was performed using SPSS17.0 software.</p><p><b>RESULTS</b>DNA binding activity of NF-κB and mRNA of IL-1β, IL-8 and COX-2 increased in cultured LPS-induced nasal epithelial cells (relative values were 1.013 ± 0.144, 0, 0, 0 respectively in control group and relative ones of LPS-induced were 2.050 ± 0.305, 1.057 ± 0.041, 0.950 ± 0.042, 0.117 ± 0.012 respectively). There was significant difference between the control group and LPS-induced nasal epithermal cells group (P values were 0.004, 0.000, 0.000, 0.000 respectively). Corresponding expression of NF-κB, IL-1β, IL-8 and COX-2 decreased after the addition of Wedelolactone (relative values were 0.917 ± 0.188, 0.180 ± 0.008, 0, 0 respectively). There was significant difference between the LPS-induced nasal epithelial cells group and the Wedelolactone-addition group (P values were 0.002, 0.000, 0.000, 0.000 respectively). But the expression of mRNA of other factors were 0 in all groups.</p><p><b>CONCLUSIONS</b>The NF-κB signal transduction pathway was involved in the transcriptional regulation of IL-1β, IL-8 and COX-2 in cultured LPS-induced nasal epithelial cells.</p>

Clinical research on the quality of life in patients with allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the quality of life (QOL) outcome in patients with allergic rhinitis (AR).</p><p><b>METHODS</b>A prospective trial was conducted to survey the QOL status of 101 AR patients, in contrast to that of 121 healthy individuals and 97 chronic pharyngitis (CP) patients by generic questionnaire medical outcomes study short-form 36-items health survey (SF-36), and to survey the most troublesome problems of AR patients by disease-specific questionnaire rhinoconjunctivitis quality of life questionnaire (RQLQ). The correlation between SF-36 and RQLQ had also been analyzed. All the results were analyzed statistically.</p><p><b>RESULTS</b>By the assessment of SF-36, the scores of 3 domains (x ± s, the same as follow, the scores were 78.02 ± 18.37, 56.13 ± 17.49, 78.81 ± 16.47, respectively) of AR patients were less than those (84.00 ± 18.36, 74.69 ± 14.13, 83.78 ± 14.31) of healthy individuals (P < 0.05), and the scores of 7 domains (the scores were: 91.78 ± 11.78, 79.16 ± 30.23, 78.02 ± 18.37, 56.13 ± 17.49, 78.81 ± 16.47, 67.66 ± 39.57, 68.78 ± 13.65, respectively) of AR patients were similar with those (94.12 ± 6.88, 80.67 ± 32.38, 73.57 ± 17.96, 59.73 ± 16.58, 80.41 ± 17.01, 63.58 ± 39.99, 66.43 ± 13.71) of CP patients (P > 0.05). By the assessment of RQLQ, in AR patients, both the nasal symptoms and the practical problems got the highest scores (the scores were 2.70 ± 1.29, 2.53 ± 1.37 respectively). According to the assessment of the correlation between SF-36 and RQLQ, the correlation was weak (r = -0.199 ∼ -0.526, P < 0.05).</p><p><b>CONCLUSIONS</b>The QOL of AR patients decreased compared with that of healthy individuals, but similar with that of CP patients. The most troublesome problems in AR patients were nasal symptoms and the practical problems. Both SF-36 and RQLQ were suitable for assessing the health status of AR patients. SF-36 and RQLQ each covered a different part of the QOL of AR patients, and the combination of the two questionnaires could improve the QOL measurement.</p>

Timing of operation and therapeutic effect of endoscopic optic nerve decompression for traumatic optic neuropathy / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To observe the optimal timing of operation and the therapeutic effect of endoscopic optic nerve decompression for traumatic optic neuropathy (TON).</p><p><b>METHODS</b>The clinical records of 90 consecutive patients with TON (93 eyes) after head and/or maxillofacial trauma from April 1998 to March 2007 were reviewed and analyzed. All patients were either unresponsive or intolerant to medication before they underwent intranasal endoscopic optic nerve decompression. The time interval between the injury and operation ranged from one day to 97 days (median 5.5 days). Among the 93 eyes, there were 71 eyes with no visual acuity before operation and 22 eyes with residue visual acuity, including light perception in 1 eye, hand movement in 5 eyes, counting fingers in 13 eyes, 0.04 in 1 eye, and 0.1 in 2 eyes. Duration of follow-up ranged from 6 days to two years (median 8 days).</p><p><b>RESULTS</b>After decompression, 35 patients (36/93 eyes, 38.7%) showed improvement of visual acuity, 53 patients (55 eyes, 59.1%) remained the same as before operation, while 2 patients (2 eyes, 2.2%) showed decreased visual acuity. Among patients with visual acuity beyond light perception before decompression, 68.2% of them (15/22 eyes) experienced visual improvement, whereas only 22.9% (8/35 eyes, 0.02 in two eyes) among patients who lost visual acuity immediately after injury, and 36.1% (13/36 eyes, 0.02 in five eyes) among those who lost visual acuity gradually after injury. There was a significant difference in visual improvement between group with visual acuity and group with no visual acuity (chi(2) = 11.864, P < 0.01). Among patients with no visual acuity, 41.2% of those (7/17 eyes) who underwent operation within 3 days of injury, experienced improvement in visual acuity, compared with 25.9% (14/54 eyes) for those who underwent the operation more than 3 days after injury. It was indicated that no significant difference in visual improvement between these two groups (chi(2) = 1.46, P > 0.05). When comparing different sites of fracture, the effect of surgery was the most desirable (55.6%, 10/18 eyes improved) if the fracture occurred simultaneously in both exterior and interior walls of optic canal, followed by the interior wall fracture (45.7%, 21/46 eyes). The operation was less effective if there was no fraction (20%, 4/20 eyes) or if the fracture occurred in exterior wall alone (11.1%, 1/9 eyes).</p><p><b>CONCLUSIONS</b>Endoscopic optic nerve decompression is a minimally invasive procedure with no adverse cosmetic effects. Early operation is recommended for saving vision, even though visual acuity is lost immediately after injury. However, the satisfactory clinical effects of endoscopic optic nerve decompression require further study.</p>

Treatment of frontal sinus diseases via combined transfrontal and intranasal endoscopic approaches / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To summarize the clinical experience of treatment of frontal sinus diseases via combined transfrontal and intranasal endoscopic approaches, and to explore its indications.</p><p><b>METHODS</b>A retrospective study of 24 patients was conducted. All patients were underwent combined transfrontal and intranasal double approaches because of different kinds of frontal sinus diseases, including 16 cases with frontal sinus cerebrospinal fluid rhinorrhea caused by traumatic comminuted fractures located in posterior wall of frontal sinus, 5 cases with osteoma, 3 cases with inverted papilloma (cancer was confirmed in one case).</p><p><b>RESULTS</b>All sixteen cases with frontal sinus cerebrospinal fluid rhinorrhea were cured after the first attempt. All cases with osteoma and inverted papilloma (including the patient with canceration) were resected completely after the first attempt. Postoperative follow-up lasted from 3 to 132 months in frontal sinus cerebrospinal fluid rhinorrhea, 8-38 months in osteoma, 7-42 months in inverted papilloma and canceration, the median follow-up period was 36 months. No recurrence was found. Operations were successful in all the patients and frontal sinus outflow tracts were unobstructed. No intraoperative or postoperative complications occurred and no disfigurement was found.</p><p><b>CONCLUSIONS</b>The potential indications for the combined transfrontal and intranasal endoscopic approaches are listed as follows, (1) frontal sinus lesions mainly located on the posterior wall, (2) propensity to recur or malignant neoplasms, (3) the lesions of frontal sinus with their lateral boundaries or operative site beyond the plane of the lamina papyracea.</p>

A case-control study of the risk factors for fungal rhinosinusitis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the risk factors of fungal rhinosinusitis.</p><p><b>METHODS</b>The preoperative clinical data of 57 patients with a diagnosis of fungal rhinosinusitis confirmed pathologically using Gomori methenamine silver staining were analyzed statistically against the data of 57 age- and gender-matched control patients with chronic rhinosinusitis.</p><p><b>RESULTS</b>Compared with chronic rhinosinusitis, fungal rhinosinusitis was characterized by a significantly shorter mean disease course (37.31 months vs 130.84 months, t = 5.59, P = 0.000). The factors related to fungal rhinosinusitis included nasal mucus, purulent nasal discharge, unilateral/bilateral sinus lesion and calcified plaque in CT scan , with odds ratios of 0.17 (0.04-0.62), 0.35 (0.15-0.80), 41 (12.50-100.00) and 91 (24.01-344.95), respectively. Conditional logistic regression identified calcified plaque in CT scan as the high-risk factor of fungal rhinosinusitis.</p><p><b>CONCLUSION</b>The presence of calcified plaque in CT scan indicates high risk of fungal rhinosinusitis and may serve as an important evidence for diagnosis of this disease.</p>

Correlation between subjective assessment and objective measurement of nasal obstruction / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the correlation between subjective assessment from clinician and patients and the objective measurement from active posterior rhinomanometry and acoustic rhinometry.</p><p><b>METHODS</b>Clinician and patients' assessment of nasal patency was achieved by visual analogue scale (VAS). Objective measurement included active posterior rhinomanometry and acoustic rhinometry. The mean of clinician's assessment and patients' VAS was compared by using paired-samples t-test. The correlation between unilateral nasal airflow resistance and unilateral nasal airway volume, unilateral minimal cross section area, and also subjective assessment and objective measurement of nasal patency were analysed by using Spearman correlation analysis in total patients.</p><p><b>RESULTS</b>In total of 316 patients, pre-decongestion and post-decongestion, unilateral nasal airflow resistance and unilateral nasal airway volume, unilateral minimal cross section area had significant negative correlation respectively (P = 0.000). The mean of clinician's assessment and patients' VAS had significant difference (P < 0.001) before and after decongestion. Clinician's assessment had significant positive correlation with patients' VAS, nasal airflow resistance, and significant negative correlation with nasal airway volume, minimal cross section area of nasal cavity before and after decongestion (P = 0.000). Patients' VAS had significant positive correlation with nasal airflow resistance, and significant negative correlation with nasal airway volume, minimal cross section area of nasal cavity before and after decongestion (P = 0.000). The correlation coefficients from clinician's assessment and objective measurements were greater than those from patients VAS and objective measurements.</p><p><b>CONCLUSIONS</b>The parameter of active posterior rhinomanometry had significant negative correlation with the parameters of acoustic rhinometry. Clinician assessment of nasal patency had significant positive correlation with patients' VAS; both of them had significant correlation with the parameters of rhinomanometry and acoustic rhinometry. Clinician's assessment was more objective and reliable to the parameters of objective measurement than patients' VAS.</p>

Transnasal endoscopic repair of nasal-skull base defect: 69 cases / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To summarize the experience of transnasal endoscopic repair of nasal-skull base defect.</p><p><b>METHODS</b>The clinical data of 69 patients with transnasal endoscopic repair of skull base defect from January 1998 to June 2006 were analyzed retrospectively. The causes, sites of defect and the effect and key technique of transnasal endoscopic repair were investigated.</p><p><b>RESULTS</b>All patients were followed-up for 6 months to 11 years. The causes were mainly traumatic fracture, occupying lesions and iatrogenic trauma. The sites were mainly in ethmoid sinus and sphenoid sinus. The success rate of one attempt was 91.3% (63/69), the ultimate success rate was 95.7% (66/69).</p><p><b>CONCLUSIONS</b>Transnasal endoscopic repair of skull base defect may be the best approach. Debriding and raw edge of defect area and "inlay-plug" method are of key technique. The liber muscle and fascia are ideal repair material.</p>

Glucocorticoid receptor subunit expression in adenoid tissue of children with obstructive sleep apnea hypopnea syndrome / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>The expression of glucocorticoid receptor (GR) expression in obstructive sleep apnea hypopnea syndrome (OSAHS) in children is currently unknown. The aim of this study was to determine the GR-alpha and GR-beta status in the adenoidal tissues in children with OSAHS.</p><p><b>METHODS</b>Thirty-four pediatric patients (aged 3-14 years, median 7.8 years) had sleep study with polysomnography before adenoidectomy. According to the criteria of apnea hypopnea index (AHI) > or = 5 /h or/and apnea index (AI) > or = 1/h, they were divided into OSAHS and non-OSAHS sub groups. The study was based on fluorescent quantitative PCR (FQ-RT-PCR) for the mRNA expression of GR-alpha and GR-beta in the adenoidal tissues in children.</p><p><b>RESULTS</b>GR isoforms mRNA encoding for expression of both GR-alpha and GR-beta were detected in the adenoids of all children. GR-alpha mRNA level [(9.40 +/- 3.06) x 10(5) cDNA copies/microg total RNA] in the adenoidal tissues in OSAHS was lower than those in the non-OSAHS [(1.60 +/- 0.26) x 10(6) cDNA copies/microg total RNA] (F = 40.285, P < 0.001), whereas no differences found for GR-beta [(1.57 +/- 0.35) x 10(4) cDNA copies/microg total RNA, (1.52 +/- 0.18) x 10(4) cDNA copies/microg total RNA]. GR-alpha/GR-beta ratio was 62.3 +/- 20. 3 in OSAHS and 107.4 +/- 24.4 in non-OSAHS. AHI or AI was not related to the mRNA levels of GR-alpha and GR-beta in OSAHS or non-OSAHS.</p><p><b>CONCLUSIONS</b>GR-alpha and GR-beta were detectable in the adenoidal tissues in children. These data indicated that the relationship between the expressions of GR and the clinical significance in OSAHS need further and profound investigation.</p>