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Objective:To investigate the effect of estrogen-related receptor α (ESRRA)-mediated lipophagy on the proliferation and migration abilities of nasopharyngeal carcinoma cells.Methods:A total of 16 clinical samples diagnosed by pathology in the Affiliated Hospital of Nantong University from 2021 to 2023 were selected, including 8 normal nasopharyngeal mucosa tissues and 8 nasopharyngeal carcinoma tissues. Immortalized normal nasopharyngeal epithelial cell line NP69 and nasopharyngeal carcinoma cell lines C666-1, CNE2, TW03-EBV and TW03 were selected. The cell lines C666-1 and CNE2 were divided into the siR-NC group (transfected with small interfering RNA negative control sequence) and siR-ESRRA group (transfected with small interfering RNA against ESRRA gene). The relative expression levels of ESRRA were detected by Western blotting and immunohistochemical assay. EdU assay was used to detect the proliferation ability of C666-1 and CNE2 cells, and Transwell assay was used to detect the migration ability. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of ESRRA and perilipin 3 (PLIN3) mRNA. The formation of lipophagy in C666-1 and CNE2 cells was observed by transmission electron microscopy. The co-localization of LC3, PLIN3 and LAMP2 with lipid droplets labeling with Bodipy was detected by immunofluorescence assay. Dual-luciferase reporter gene assay was used to verify the targeting relationship between ESRRA and PLIN3.Results:The relative expression level of ESRRA in nasopharyngeal carcinoma tissues was higher than that in normal nasopharyngeal mucosa tissues(1.15±0.75 vs. 0.32±0.21, t = 3.02, P = 0.009). The relative expression level of ESRRA in nasopharyngeal carcinoma cell lines C666-1 (1.539±0.044), CNE2 (1.420±0.030), TW03-EBV (2.867±0.044), and TW03 (1.323±0.022) were higher than that in normal nasopharyngeal epithelial cell line NP69 (0.094±0.002), and the difference was statistically significant ( F = 34.08, P < 0.001).The results of EdU assay showed that the proportions of EdU labeled positive cells in CNE2 cells of siR-NC group and siR-ESRRA group were (70.44±4.06)% and (51.51±0.92)% ( t = 7.88, P = 0.001), and the proportions in C666-1 cells were (62.25±3.89)% and (54.91±0.27)% ( t = 3.26, P = 0.031). The results of Transwell assay showed that the number of migrating cells in CNE2 and C666-1 cells was less than that in siR-NC group [CNE2 cells: (181±7) cells vs. (261±21) cells; C666-1 cells: (201±16) cells vs. (256±7) cells], and the differences were statistically significant ( t = 6.30, P = 0.003; t = 5.43, P = 0.006). According to qRT-PCR results, the relative expression level of PLIN3 mRNA in the siR-ESRRA group was higher than that in the siR-NC group (CNE2 cells: 1.58±0.16 vs. 0.83±0.17, t = 5.59, P = 0.005; C666-1 cells: 1.37±0.12 vs. 1.06±0.06, t = 3.86, P = 0.018). The dual-luciferase reporter gene assay results indicated a targeted binding interaction between PLIN3 and ESRRA. Transmission electron microscopy observation showed that the lipid droplets in nasopharyngeal carcinoma cells increased and the binding to autophagosomes decreased after knockdown of ESRRA. The results of immunofluorescence assay demonstrated that, in contrast to the siR-NC group, there was a decrease in the co-localization of LC3 and LAMP2 and an increase in the co-localization of lipid droplets with PLIN3. Conclusions:ESRRA is highly expressed in nasopharyngeal carcinoma tissues and cells. As a transcription repressor, ESRRA may work to prevent PLIN3 from being transcribed, decrease lipid droplet stability, mediate lipophagy, and promote proliferation and migration of nasopharyngeal carcinoma cells.
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<p><b>OBJECTIVE</b>The study was designed to analyze the efficacy of standardized house dust mite allergen vaccine to allergic rhinitis by subcutaneous injection and investigate the possible mechanism of specific immunotherapy (SIT).</p><p><b>METHODS</b>From January 2011 to December 2011 a prospective study was performed in the Department of Otorhinolaryngology Head and Neck Surgery, the Affiliated Hospital of Nantong University, involving 90 patients with perennial AR, of whom 60 patients received Der p - SIT + pharmacotherapy after their approval and 30 received only pharmacotherapy. All patients were allergic to house dust mites. Symptom and medication scores were recorded three times: before the treatment, at the middle of treatment and at the end of treatment. Over a period of 1 yr. prior to and at the end of treatment, CD4⁺ CD25⁺ Foxp3⁺ Treg cells and Th17 cells were measured by flow cytometry. SPSS 21.0 software was used to analyze the data.</p><p><b>RESULTS</b>The symptom scores using VAS and medication scores in AR patients treated with SIT and medication were reduced, the differences were significant (14.25 ± 6.40, 1.00 ± 0.84 vs. 32.18 ± 7.78, 3.12 ± 1.54, t value was 19.65, 10.71, both P < 0.05). The symptom of VAS score in medication group was reduced after treatment (30.30 ± 5.97 vs. 20.30 ± 5.79, t = 10.09, P < 0.05), but the medication score had not significant difference (P > 0.05). The frequency of Th17 cells in peripheral blood mononuclear cells were decreased in patients treated with SIT, whereas the frequency of Treg cells were increased (χ² value was 2.81, 2.80, both P < 0.05), but not in medication group.</p><p><b>CONCLUSIONS</b>Both SIT and pharmacotherapy can improve symptoms of allergic rhinitis, but SIT can also reduce medication use. The effect of immunotherapy is better than drug treatment alone. The frequency of blood Th17 cells in peripheral blood mononuclear cells were decreased in patients treated with SIT, whereas the frequency of Treg cells were increased.</p>
Subject(s)
Animals , Humans , Antigens, Dermatophagoides , Immunotherapy , Leukocytes, Mononuclear , Prospective Studies , Pyroglyphidae , Rhinitis, Allergic , Therapeutics , T-Lymphocytes, Regulatory , Cell Biology , Th17 Cells , Cell Biology , Vaccines , Therapeutic UsesABSTRACT
Objective To explore the health-relate quality of life(QOL) status of adult persistent allergic rhinitis (PAR);the change of QOL of pro-post specific immunotherapy (SIT) and pharmacotherapy .Methods Skin prick tests(SPT) were performed on PAR patients .According to the results ,80 adult cases that were allergic to dermatophagoides were enrolled in ENT outpatient clinic of affiliated hospital of Nantong University from April to August 2011 .The patients were randomly allocated to receive either specific immunotherapy(n=40) or pharmacotherapy (n=40) ,all of them were given RQLQ before and after half-year treatment ;40 cases without any allergic diseases were chosen from ENT in-patient department ,and were given RQLQ .The scores of previous treatment of the PAR group were compared with health control group ,then compared with the scores of post-treatment ,and also compared the scores of post-treatment between the immunotherapy group and pharmacotherapy group .Results The scores of the PAR patients were higher than that of health control patients in all dimensions of RQLQ (P< 0 .05) ,and the most troublesome problems were nasal symptoms .The scores of the patients who received SIT were evidently lower than that of pro-treatment in all dimensions of RQLQ(P<0 .05) ,the scores of the patients who received medical treatment were also lower than before (P<0 .05) , and the scores of the SIT group were lower than the pharmacotherapy group (P<0 .05) .Conclusion The QOL of adult patients with PAR was improved after SIT or drug treatment ,and QOL improvement is more obvious treat by SIT .
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Objective To investigate the impact of different anesthesia methods on intraoperative and postoperative patients un-dergoing laparoscopic rectal cancer surgery and to explore the ideal anesthetic method for laparoscopic rectal cancer surgery .Meth-ods 40 cases of laparoscopic resection for rectal cancer ,ASAⅠ-Ⅱ grade ,were selected and randomly divided into two groups .The group A (20 cases) was performed general anesthesia combined with epidural anesthesia and the B group (20 cases) was performed systemic anesthesia .The intraoperative hemodynamics ,respiratory function ,awaking time and awaking quality evaluation were ob-served .Results The airway pressure(Paw ) and PETCO2 in the two groups were increased .The intraoperative hemodynamics in the group A was more stable than those in the group B .The awaking time and awaking quality in the group A were superior to those in the B group .Conclusion Compared with simple general anesthesia ,general anesthesia combined with epidural anesthesia for laparo-scopic rectal cancer surgery has more stable vital signs and better awaking quality ,whoich is an ideal anesthetic method for laparo-scopic rectal cancer surgery .
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AIM:To examine the expression profiles of both genes and proteins in hippocampus of rats with temporal lobe epilepsy(TLE)for revealing the molecular mechanisms of TLE and looking for the candidate targets and new therapeutic approaches in clinical practice.METHODS:Rat temporal lobe epilepsy was induced by administration of lithium chloride and pilocarpine(LiCl-PILO).The expression spectra of genes and proteins were constructed through the techniques of cDNA microarray,two-dimensional(2D)electrophoresis and Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS).Subsequently,the differentially expressed genes and proteins were identified and analyzed.RESULTS:There were 192 genes of differential expression observed in hippocampal tissues of LiCl-PILO-induced temporal lobe epilepsy,and 159 genes have been registered in Genbank database,in which 84 genes were up-regulated while 75 genes were down-regulated.78 protein spots of differential display were screened out,in which 31 proteins were detected to be down-regulated and 47 were up-regulated.Finally,5 proteins were identified.CONCLUSION:These genes and proteins found in our study may play pivotal roles in the pathogenic mechanisms of epilepsy and may promise new therapeutic targets for refractory epilepsy in the future.