Correlation of IL-19 and its receptors with tissue remodeling in chronic rhinosinusitis / 中国病理生理杂志

AIM:To explore the differential expression of interleukin-19 (IL-19) and its receptors (IL-20R1/IL-20R2) in the samples of chronic rhinosinusitis (CRS) with or without nasal polyps (CRSwNP and CRSsNP), and to investigate the correlation of IL-19 and its receptors with tissue remodeling factors, including matrix metalloproteinase (MMP)-2, MMP-9 and tissue inhibitor of metalloproteinase (TIMP)-1 in CRS.METHODS:The polyps from CRSwNP patients (n=30), the sinus mucosa from CRSsNP patients (n=15) and the inferior turbinal mucosa of nasal septum de-viation patients (n=15) were collected.The expression of IL-19 and its receptors (IL-20R1/IL-20R2) in each group was detected by real-time RT-PCR and immunohistochemistry.The expression of tissue remodeling factors MMP-2, MMP-9 and TIMP-1 in different groups was detected by real-time RT-PCR and ELISA.RESULTS:Increased mRNA and protein expression levels of IL-19, IL-20R1, IL-20R2 and MMP-9 were observed in CRSwNP group compared with CRSsNP and control group (P<0.05), while elevated expression level of TIMP-1 was observed in CRSsNP group compared with CRSwNP and control group (P<0.05).The relative expression of MMP-2 among the 3 groups showed no obvious difference.The expression of IL-19 and its receptors showed significantly positive correlations with MMP-9 in CRSwNP (P<0.05).No significant correlation between IL-19 and/or its receptors with TIMP-1 in CRSwNP group was observed.CONCLUSION:The high expression levels of IL-19, IL-20R1 and IL-20R2 and their positive correlations with MMP-9 in CRSwNP indica-te that IL-19 and its receptors may be involved in the tissue remodeling of chronic rhinosinusitis.

Glucocorticoid receptors mRNA expression in peripheral blood mononuclear cell in patients with obstructive sleep apnea hypopnea syndrome / 中国耳鼻咽喉头颈外科

[ABSTRACT]OBJECTIVETo study the mRNA expression of glucocorticoid receptor(GR) in peripheral blood mononuclear cells in patients with obstructive sleep apnea hypopnea syndrome(OSAHS) and its clinical significance. METHODSReal-time fluorescent quantitative PCR was used to detect GRα mRNA and GRβ mRNA exprssion in PBMC of 30 male patients with moderate to severe OSAHS and 27 healthy male subjects. The relationships between the expression of glucocorticoid receptor mRNA and the apnea hypopnea index, body mass index, neck circumference, waist circumference, the lowest oxygen saturation, the average oxygen saturation, the Epworth sleepiness scale(ESS) score, fasting blood glucose, heart rate and blood pressure were analyzed.RESULTSCompared with the control group, the expression level of GRα mRNA was lower in the OSAHS group(t=2.25,P0.05). We had not found the significant correlation between the expression of GRα mRNA and the clinical parameters above in OSAHS patients.CONCLUSIONThe expression of GRα mRNA in PBMC in moderate to severe OSAHS male patients have a downward trend compared with healthy group, but the mechanism remains unclear.

The preparation of recombinant adenovirus Ad-Rad50-GFP and detection of the optimal multiplicity of infection in CNE1 transfected hv Ad-Rad50-GFP / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied.@*METHOD@#The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection.@*RESULT@#The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours.@*CONCLUSION@#Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.

Effects of tetrandrine on enhancing radiosensitivity of human nasopharyngeal carcinoma cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the effect and mechanism of tetrandrine (Tet) on enhancing radiosensitivity of human nasopharyngeal carcinoma cell lines in vitro.</p><p><b>METHODS</b>CNE1 and CNE2 were exposed to radiation with or without Tet, the DNA damage of the cells were evaluated by neutral comet electrophoresis, and cell cycle and apoptosis were analyzed by flow cytometry.</p><p><b>RESULTS</b>The mean tail movements (TM) of CNE1 treated with radiation or radiation plus Tet were (7.13 ± 3.70) (X(-) ± s) and (13.61 ± 5.45), respectively (t = 2.784, P < 0.05), and TM of CNE2 treated with radiation or radiation plus Tet were (11.52 ± 4.04) and (18.85 ± 6.18), respectively (t = 3.089, P < 0.05). With the exposure to radiation or radiation plus Tet, the percentages of CNE1 in G2 phases were (42.62 ± 2.07)% and (17.02 ± 1.87)%, respectively (t = 23.173, P < 0.01), and the percentages of CNE2 in G2 phases were (34.82 ± 2.74)% and (19.64 ± 4.82)%, respectively(t = 16.500, P < 0.01). There was no significant difference in the apoptosis rates between the cells treated with radiation or radiation plus Tet regardless of CNE1 (17.24 ± 0.99)% vs (19.11 ± 1.24)%, and CNE2 (16.68 ± 0.27)% vs (18.51 ± 2.41)% (P > 0.05).</p><p><b>CONCLUSIONS</b>Tet can enhance radiosensitivity of human nasopharyngeal carcinoma cell lines. The mechanism could be related to abrogation of radiation-induced G2/M arrest and reduction of double-strand break repair capacity.</p>

Effects of lentivirus-mediated epidermal growth factor-like domain 7 silencing on proliferation and invasion of human laryngeal carcinoma Hep-2 cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the effects of epidermal growth factor-like domain 7 (EGFL7) gene silencing on the proliferation and invasion ablity of laryngeal carcinoma cells.</p><p><b>METHODS</b>A lentiviral vector expressing EGFL7 shRNA was constructed and transfected into human laryngeal carcinoma Hep-2 cells. The expressions of EGFL7 mRNA and protein were detected by Real-time PCR and Western blot, respectively. Cell proliferation was evaluated by CCK-8 assay, cell cycle and apoptosis were tested by flow cytometry, and cell invasion was detected by transwell invasion assay.</p><p><b>RESULTS</b>The relative expression level s of EGFL7 mRNA and protein in EGFL7-SuRNA group were svgnificantly lower than control group (P < 0.001). Western blot analysis proved that the relative expression of EGFL7 protein in NC group, Lenti-NC group and Lenti-EGFL7 group was (0.39 ± 0.12),(0.36 ± 0.14) and (0.07 ± 0.04), respectively. EGFL7 expression in Lenri-EGFL7 group was significantly inhibited than NC group (P < 0.001), which confirmed that the recombinant lentivirus was successfully transfected into Hep-2 cells. The proliferation of Hep-2 cells was significantly inhibited after transfection (P < 0.01). Compared with the NC group and Lenti-NC group, the proportion of cells in S phase was significantly increased in Lenti-EGFL7 group (P < 0.01), and the proportion in G1 phase was significantly decreased (P < 0.05). Cell apoptosis assay showed that the apoptotic rate in Lenti-EGFL7 group (66.2 ± 1.28) % was significantly increased in NC group (6.09 ± 3.28) % and Lenti-NC group (9.86 ± 2.13) %. In Transwell invision assay, the mean number of cells coming through the Metrigel in Lenti-EGFL7 group was significantly decreased than in the NC group and Lenti-NC group (P < 0.01).</p><p><b>CONCLUSION</b>The proliferation and invasion ablity of laryngeal carcinoma Hep-2 cells can be inhibited by siRNA mediated EGFL7 gene silencing.</p>