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Objectives@#. Reactive oxygen species in the stria vascularis (SV) of the cochlea may be involved in the pathogenesis of sensorineural hearing loss. However, the effects of oxidative stress on SV endothelial cells (SV-ECs) remain largely unknown, and no feasible in vitro cell culture model exists for the functional study of SV-ECs. @*Methods@#. We isolated primary SV-ECs from the SV of neonatal mice. The apoptosis-reducing effects of fibronectin in SV-ECs cultured with serum-free medium were determined using β-galactosidase staining and flow cytometry. SV-ECs incubated in serum-free medium were treated with various H2O2 concentrations to evaluate the effects of H2O2 on their viability. The secretome of SV-ECs treated with or without H2O2 (100 μM or 500 μM) was analyzed using high-resolution mass spectrometry. The function of the SV-EC secretome was evaluated by a macrophage assay. @*Results@#. We successfully isolated and characterized the SV-ECs. Treatment with H2O2 at concentrations up to 500 μM for 2 hours and further incubation with serum-free medium in plates precoated with fibronectin showed no significant effect on apoptosis. Compared to the control SV-ECs, the amount of differential proteins in the secretome of SV-ECs stimulated with 500 μM H2O2 was much higher than in those treated with 100 μM H2O2. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses suggested that the proteins differentially expressed in SV-ECs treated with 500 μM H2O2 were involved in the regulation of multiple signaling pathways and cellular processes. The secretome of H2O2-stimulated SV-ECs exhibited significant pro-inflammatory effects on macrophages. @*Conclusion@#. We successfully established an in vitro serum-free culture method, identified the differential proteins released by oxidative stress-induced ECs and their functions, and revealed the pro-inflammatory effects of the secretome of H2O2-stimulated SV-ECs. Therefore, SV-ECs might elicit immunoregulatory effects on bystander cells in the microenvironment of oxidative stress-induced cochlea, especially cochlear macrophages.
ABSTRACT
[Objective] To evaluate the efficacy of sound therapy and provide a kind of treatment for tinnitus patients with normal audiogram.[Methods] From the database of the Audiometric Center of the First Affiliated Hospital,Sun Yat-sen University,we identified 46 outpatients with normal hearing thresholds in the conventional pure-tone audiometry suffering subjective tinnitus over 3 months.The clinical information were collected with respect of age,gender,tinnitus severity,pitch,laterality and duration,comorbid symptoms.All the patients received the tinnitus test,including loudness and frequency of the tinnitus and residual inhibition,Tinnitus Handicap Inventory (THI) and Visual Analogue Scale (VAS) scores pre-and post-complex sound therapy.The complex sound were remixed with natural sounds and music materials,while the masking frequency of natural sounds is consistent with the tinnitus frequency of patients and the main frequency of music materials covers the tinnitus frequency.[Results] 32 of the 46 patients showed residual inhibition (RI) positive.The average THI score before treatment was 33.38±16.23 and the VAS score was 4.38±1.62.The THI score decreased to 21.75±11.67 and the VAS score was 2.97±1.06 after sound therapy.The rest 14 patients were residual inhibition (RI) negative.The THI score were 29.86±20.15 and 23.43±16.29 pre-and post-complex sound therapy,while the VAS score were 3.93±1.69 and 3.36±1.2.The THI and VAS scores were significantly decreased after the treatment in both RI positive and negative patients.The RI positive group showed better results than RI negative group.[Conclusions] Both RI positive and negative tinnitus with normal audiometry shown adaptation with complex sound therapy,RI positive patients acquired more benefits from tailor-made complex sound therapy.
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<p><b>OBJECTIVE</b>To detect the expression of DJ-1 in laryngeal squamous cell carcinoma (LSCC) and to study the relationship between DJ-1 expression and clinical indexes of LSCC.</p><p><b>METHODS</b>The expressions of DJ-1 protein in 71 LSCC samples and 9 cases control samples from laryngeal mucosa tissues of non-LSCC patients were detected using streptavidin peroxidase immunohistochemistry staining and the relationships between DJ-1 protein expression and clinicopathologic characteristics were analyzed.</p><p><b>RESULTS</b>(1) The positive expression rate of DJ-1 protein in LSCC was 85.9%(61/71), which was significantly higher than the rate (55.5%, 5/9) in control laryngeal mucosa tissues (P < 0.05). (2) DJ-1 expression was related to tumor recurrence (P < 0.05), but not to sex, age, primary cancer position, T stage, clinical stage, lymph node metastasis and tumor differentiation. Tumor recurrence rate (53.3%) in the patients with higher expression of DJ-1 protein was higher than the rate (26.8%) in the patients with lower expression of DJ-1 protein (χ(2) = 5.164, P < 0.05). (3) With Kaplan-Meier curves and Cox regression analysis, the cumulative 5-year survival rates were correlated with DJ-1 expression levels in laryngeal cancer tissues or cervical lymph node metastasis (all P < 0.05), but not to sex, age, primary cancer position, T stage, clinical stage and tumor differentiation.</p><p><b>CONCLUSIONS</b>The expression of DJ-1 protein in LSCC is higher than that in control laryngeal mucous tissues. Overexpression of DJ-1 is associated with poor overall survival in LSCC patients.</p>