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Yan W.,Shandong University | Yan W.,Shandong Institute of Otolaryngology | Li J.,Shandong University | Li J.,Shandong Institute of Otolaryngology | And 11 more authors.
PLoS ONE | Year: 2014

Objectives: In this study, using an Streptococcus pneumoniae-induced tympanosclerosis (TS) model, we explored the effects of captopril and losartan in the treatment of TS and the possible mechanisms. Study Design: A prospective experimental animal study. Methods: We set up the TS models in both guinea pig and wistar rat by inoculation of type-3 Streptococcus pneumoniae microorganisms and then treated the animals with the combining use of captopril and losartan. Otomicroscopy was employed to observe the development of TS. Auditory brainstem response was used to test the hearing function of animals. Hematoxylin-eosin and von Kossa staining were performed to determine the morphological changes and calcium depositions. The protein expressions of transforming growth factor β1 (TGF-β1) were assessed by western blot and immunohistochemistry staining, and the mRNA level of TGF-β1 was measured by quantitative reverse transcription-polymerase chain reaction. Results: The combining use of captopril and losartan attenuated TS responses in terms of a decrease in the TS incidence and the ABR threshold, a reduction of hyalinization and calcification in the middle ear mucosa and the thickness of the mucosa. In addition, the TGF-β1 expression was decreased at both protein and mRNA levels. Conclusion: Our data indicate, for the first time, that the combining use of captopril and losartan obviously attenuates TS progress through inhibiting the overexpressing of TGF-β1. © 2014 Yan et al. Source


Guo W.,Shandong University | Bai X.,Shandong University | Bai X.,Shandong Provincial Key Laboratory of Otology | Han Y.,Shandong University | And 8 more authors.
Laryngoscope | Year: 2012

Objectives/Hypothesis: The present study was performed to investigate the expressions of transforming growth factor β1 (TGF-β1) and matrix metalloproteinase-9 (MMP-9) in an experimental model of tympanosclerosis and their possible roles in the formation of this disorder. Study Design: A prospective experimental animal study. Methods: Seventy guinea pigs were used in this study, of which 10 were chosen to serve as controls, and the other 60 were used in the tympanosclerosis group by inoculation of type-3 Streptococcus pneumoniae microorganisms. The experimental animals were further divided into six subgroups on the basis of six time points. Otomicroscopy was employed to observe the development of myringosclerosis. Hematoxylin-eosin and von Kossa staining were performed to determine the morphological changes and calcium depositions. The expressions of TGF-β1 and MMP-9 were assessed by Western blot and immunohistochemistry. Results: Slight sclerotic changes in tympanic membrane were found at week 2, and extensive myringosclerosis was observed at week 6. Hyalinization and calcification in the tympanic membrane and middle ear mucous membrane were clearly visible at week 6. Expression of TGF-β1 was significantly increased with the development of tympanosclerosis. Expression of MMP-9 was increased from week 1 to week 4, and then declined at week 6. These two cytokines were both distributed in the cytoplasm of fibroblast cells and inflammatory cells, which were widely distributed in the tympanic membrane and middle ear mucosa at week 6. Conclusions: Our data indicate that, for the first time, the alteration in expressions of TGF-β1 and MMP-9 were involved in the formation of tympanosclerosis, which may represent an important mechanism underlying the pathogenesis of tympanosclerosis. Copyright © 2012 The American Laryngological, Rhinological, and Otological Society, Inc. Source


Wang J.,Shandong University | Li X.,Shandong University | Li X.,Shandong Provincial Key Laboratory of Otology | Zhang Z.,University of Pennsylvania | And 4 more authors.
Neuroscience Letters | Year: 2015

Sperm-associated antigen 6 (Spag6) gene, which encodes an axonemal protein (SPAG6), ubiquitously expresses in tissue and organs containing ciliated cells. The present work was to investigate whether SPAG6 expressed in cochlear hair cells and, if so, to explore the presumable correlations between prestin and SPAG6. The distribution of SPAG6 in organ of Corti and the morphological features of hair cells in basilar membrane were investigated by immunofluorescent staining. The amount of prestin in Spag6 mutant mice was measured by Western blotting and real-time PCR, respectively. Additionally, co-immunoprecipitation tests were performed to confirm the presumed interaction between prestin and SPAG6. We observed that SPAG6 expressed in the cuticular plate in outer hair cells (OHCs) and prestin in the lateral wall of OHCs that located along with SPAG6 at this site. In comparison to Spag6 +/+ mice, Spag6 -/- mice showed apparent morphological abnormity of OHCs and lower intensity of prestin fluorescence. The expression of prestin in Spag6 -/- mice reduced significantly at both protein and mRNA levels. Moreover, co-immunoprecipitation tests demonstrated the interaction between prestin and SPAG6. Taken together, these data indicate that SPAG6 is indispensible for the stability of OHCs by maintaining the normal expression of prestin, which implies that Spag6 gene is essential for mechanosensory function of OHCs. © 2015 Elsevier Ireland Ltd. Source


Bai X.,Shandong University | Bai X.,Shandong Provincial Key Laboratory of Otology | Lv H.,Shandong University | Zhang F.,Shandong University | And 13 more authors.
American Journal of Medical Genetics, Part A | Year: 2014

Hearing loss is the most common sensory disorder affecting 278 million people in the world, and more than 60% of hearing loss patients can be attributed to genetic causes. Although many loci have been linked to hereditary hearing impairment, most of the causative genes have not been identified as yet. The goal of this study was to investigate the cause of dominantly inherited sensorineural all-frequency hearing loss in a six-generation Chinese family. We performed exome sequencing to screen responsible candidate genes in three family members with all-frequency hearing loss and one member with normal hearing. Sanger sequencing was employed to examine the variant mutations in the members of this family and 200 healthy persons. PCR-RFLP was performed to further confirm the nucleotide mutation. A novel missense mutation c.2389G>A (GAC→AAC) in WFS1 gene was identified, which was co-segregated with the hearing loss phenotype. No mutation was found in 200 controls and the family members with normal hearing in this site. The present study identifies, for the first time, a novel mutation in WFS1 gene that causes non-syndromic hearing loss in all, rather than in low or high, frequencies. © 2014 Wiley Periodicals, Inc. Source


Bai X.,Shandong University | Bai X.,Shandong Provincial Key Laboratory of Otology | Wang D.,Shandong University | Fan Z.,Shandong University | And 11 more authors.
B-ENT | Year: 2012

Objectives: To evaluate the presence of Helicobacter pylori (HP) in middle ear effusion from Chinese adult patients with otitis media with effusion (OME). Methodology: A total of 60 aspiration samples collected from adult patients (ranging from 19 to 73 years old) were included in the study. The effusion samples aspirated from the patients' middle ears were analyzed with polymerase chain reaction (PCR) assay and cultured for 7 days under microaerobic conditions. Subsequently, Gram staining and urease testing were performed to identify HP. Results: Of the 60 middle ear effusion samples, 24 (40%) were shown to be HP-positive by PCR assay. Of these, seven specimens were confirmed HP-positive by both culture and urease test. Conclusions: The present study clearly demonstrates the presence of HP in the middle ear effusion of some Chinese adult patients with OME. This finding suggests that HP may be involved in the etiology of OME. Additional studies are needed to investigate the exact role of HP in the pathogenesis of OME. Source

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