Expression and Correlation Research of MicroRNA10a-5p and PIK3CA in Middle Ear Cholesteatoma.

BACKGROUND: This study aimed to examine the roles of miR-10a-5p and phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α in the pathogenesis of middle ear cholesteatoma. METHODS: We enrolled 27 patients with middle ear cholesteatoma and collected samples of intraoperative cholesteatoma and normal posterior ear skin tissues. The mRNA expression levels of miR-10a-5p and PIK3CA were detected using real-time quantitative polymerase chain reaction. PIK3CA protein expression was measured by immunohistochemistry and western blotting. RESULTS: Middle ear cholesteatoma tissues showed significantly lower miR-10a-5p expression levels and significantly higher PIK3CA expression levels than normal posterior ear skin tissues (both P < .05). Furthermore, the miR-10a-5p and PIK3CA expression levels were significantly negatively correlated in middle ear cholesteatoma tissues (r = -0.926, P < .001). CONCLUSION: Low miR-10a-5p expression levels in middle ear cholesteatoma tissues may inhibit the growth and proliferation of cholesteatoma, whereas high PIK3CA expression level may promote its growth and proliferation. In addition, miR-10a-5p may affect the proliferation and differentiation of cholesteatoma by negatively regulating its target gene, PIK3CA.

Dynamic Changes, Correlation of Basophils, and the Therapeutic Effect in Patients With Allergic Rhinitis During Allergen-specific Immunotherapy.

BACKGROUND: The role of basophils in allergic rhinitis (AR) has been studied extensively; however, there are very few reports on changes in basophils after allergen-specific immunotherapy (SIT). OBJECTIVE: To examine the changes and correlation of peripheral blood basophils and the therapeutic effect in patients with AR during allergen-SIT. METHODS: A total of 77 patients with AR who were allergic only to house dust mites received allergen-SIT. At 3 time points, patients underwent testing for the percentage and activation rate of basophils in peripheral blood, skin index (SI) measurement, visual analog scale (VAS) assessment, and rhinoconjunctivitis quality of life questionnaire (RQLQ) evaluation. The results were compared to a control group with congenital preauricular fistula. RESULTS: (1) Before treatment, the percentage and activation rate of basophils in patients with AR were significantly higher than those in controls. There was no significant difference in the percentages and activation rates of basophils at the 3 time points. (2) The SIs, VAS, and RQLQ scores of the patients immediately after treatment and 2 years posttreatment decreased significantly compared to those before treatment; the SI, VAS, and RQLQ scores of the patients 2 years posttreatment increased significantly compared with those immediately after treatment. (3) There was no correlation between the patients' basophil activation rate and percentage and the SI, VAS, and RQLQ scores at all time points. CONCLUSION: The percentage and activation rate of basophils were higher in patients with AR than in controls. The values did not change significantly after allergen-SIT and showed no correlation with treatment effectiveness. Therefore, the frequency and activation rate of basophils cannot be used as criteria for assessing the effectiveness of allergen-SIT for house dust mites. Allergen-SIT is effective for the management of AR, but the effect declines after the completion of therapy.

Migratory fishbones in the pharynx: A report of two cases.

The possibility of fishbone migration into the surrounding tissues, especially in cases where it cannot be identified on routine inspection. Early diagnosis of migratory fishbone and therapeutic management are essential for optimal patient survival.

[Analysis of related factors between sudden sensorineural hearing loss and serum indices base on artificial intelligence and big data].

Objective:The etiology and pathophysiologic mechanism of sudden sensorineural hearing loss are undefined. We will use artificial intelligence and big data methods to explore the correlation between sudden sensorineural hearing loss and serum indices. Method:A total of 1218 patients with sudden deafness admitted to Sun Yat-sen Memorial Hospital were selected as the experimental group, 95 861 healthy subjects were randomly selected as the control group at the same period. Serum biochemical indexes in two groups were collected and analyzed by TreeNet and CART machine learning algorithms, to screen out highly correlated indicators with sudden sensorineural hearing loss and dig out a set of clinical features for people with high risk of sudden sensorineural hearing loss. Result:It was found that high prevalence rate of sudden sensorineural hearing loss is related to eosinophils, reticulocyte and fibrinogen. The areas under the receiver operator characteristic curves（ROC-AUC） were exploited to evaluate the prediction performance of TreeNet model. Overall the TreeNet model has provided high predictive ability by ROC curve, achieving AUC of 0.99, both recall and accuracy rate of 99.90%. Conclusion:There is significant difference between sudden deadness and normal people in serum biochemical indexes. Eosinophil is the first important indicator to distinguish sudden sensorineural hearing loss. Treenet model has important referenced significance for the screening and diagnosis of sudden sensorineural hearing loss.

Missed hearing loss in tinnitus patients with normal audiograms.

The prevalence of tinnitus is positively correlated with hearing loss, although, tinnitus can also present alongside clinically normal pure-tone thresholds. As standard pure tone audiograms (PTA) only sample at octave or inter-octaves, they potentially can miss lesions between the tested frequencies. Here we investigate if tinnitus patients with normal audiograms have hearing loss missed by standard PTA testing, referred as "missed hearing loss" in the paper. Hearing thresholds in 106 tinnitus patients who have a normal PTA were tested using fine frequency resolution (1/24 octave step) audiometry, referred as precision PTA (P-PTA), at ±1/3 octave band centered at their tinnitus frequencies. Tinnitus pitch, loudness and residual inhibition were evaluated based on the result of P-PTA. DPOAEs were also tested to evaluate the function of outer hair cells (OHC). Using the P-PTA test, we found that 49% (52 out of 106) of tinnitus patients with normal audiograms showed sharply notched hearing loss and most of the notches were at their tinnitus frequencies. Using a fine frequency step (1/24 octave) to assess tinnitus pitch, the successful matching rate increased to 84%, significantly higher than the rate measured in traditional method (51%, Fisher's test, P < 0.0001, n = 106). The number of patients whose tinnitus loudness was less than 5 dB SL increased from 26% to 55% after the loudness reassessment based on the tinnitus pitch match (n = 106, Chi-Square test, P < 0.01). The percentage of patients who showed positive residual inhibition of tinnitus also increased from 31% to 54% (Chi-Square test, P < 0.05). DPOAEs revealed significantly reduced OAE amplitude in the tinnitus patients, suggesting that OHC dysfunction may contribute to their notched hearing loss. However, in 13 out of 31 patients with notched hearing loss, their DPOAE did not show any reduction which suggests that their notched hearing loss may be induced by the dysfunction of the inner hair cells or afferent synapses (synaptopathy). Our study confirmed hidden cochlear impairments in tinnitus patients with seemingly normal audiograms. We conclude that P-PTA can help identify a mild hearing impairment that may otherwise be missed by conventional PTA and that P-PTA can also improve tinnitus evaluation accuracy.

Diagnostic values of serum cathepsin B and D in patients with nasopharyngeal carcinoma.

BACKGROUND: The diagnostic and prognostic significance of increased cathepsin B (CTSB) and cathepsin D (CTSD) concentration in the serum of cancer patients were evaluated for some tumor types. High expression of CTSD and CTSB was detected in biopsy tissues from nasopharyngeal carcinoma (NPC). However, whether CTSD and CTSB serve as diagnostic and prognostic markers of NPC remains unclear. METHODS: Serum samples were collected from 40 healthy volunteers and 80 NPC patients enrolled in the study. CTSB and CTSD in the serum samples were detected using enzyme-linked immunosorbent assay (ELISA). Concomitantly, the relationship between CTSB and CTSD concentrations and clinicopathological prognosis was assessed. The sensitivity and specificity of the two components in the diagnosis of NPC were evaluated in 80 NPC patients. RESULTS: ELISA analysis showed that in the sera obtained from NPC patients, the CTSB concentration was 12.5 ± 3.5 mg/L (median, 12.4 mg/L), and the CTSD concentration was 15.7 ± 8.7 mg/L (median, 14.7 mg/L). CTSB and CTSD levels were significantly higher in the NPC patient population compared to the healthy control population (p = 0.001; p = 0.001, respectively). The presence of CTSB and CTSD in the serum of the patients with NPC correlated with the tumor node metastasis (TNM) scores (p = 0.001). Other parameters were not identified to be of significance. Receiver operating characteristic (ROC) analysis showed that a cut off CTSB concentration of 12.4 mg/L had 61.9% sensitivity and 63.2% specificity in the prediction of progression-free survival (Area under the curve (AUC) = 0.525; 95% CI, 39.7-65.2; p = 0.704); whereas a cut off CTSD concentration of 14.7 mg/L had 66.7% sensitivity, and 58.5% specificity (AUC = 0.552; 95% CI, 42.3-68.1; p = 0.42). CONCLUSIONS: Serum CTSB and CTSD concentrations were found to have a diagnostic value in NPC. However, the CTSB and CTSD serum levels had no prognostic role for the outcome in NPC patients.

Bone marrow-derived mesenchymal stem cells differentiate into nerve-like cells in vitro after transfection with brain-derived neurotrophic factor gene.

Bone marrow-derived mesenchymal stem cells can differentiate into a variety of adult cells. Brain-derived neurotrophic factor (BDNF) is briefly active during differentiation and induces mesenchymal stem cells to differentiate into nerve cells. In this study, we cloned human BDNF to generate a recombinant pcDNA3.1(-)-BDNF vector and transfected the vector into bone marrow-derived mesenchymal stem cells. We selected these cells with Geneticin-418 to obtain BDNF-BMSCs, which were induced with retinoic acid to obtain induced BDNF-BMSCs. The transfected cells displayed the typical morphology and surface antigen profile of fibroblasts and were observed to express clusters of differentiation 29, 44, and 90 (observed in matrix and stromal cells), but not clusters of differentiation 31, 34, and 45 (observed in red blood cells and endothelial cells), via flow cytometry. Enzyme-linked immunosorbent assays showed that transfected bone marrow-derived mesenchymal stem cells secreted more BDNF than non-transfected bone marrow-derived mesenchymal stem cells. Immunocytochemistry and real-time reverse transcription polymerase chain reaction analysis showed that non-induced BDNF-BMSCs maintained a higher proliferative capacity and expressed higher amounts of brain-derived neurotrophic factor, nestin, neuron-specific enolase, and glial fibrillary acid protein than non-transfected bone marrow-derived mesenchymal stem cells. An additional increase was observed in the induced BDNF-BMSCs compared to the non-induced BDNF-BMSCs. This expression profile is characteristic of neurocytes. Our data demonstrate that bone marrow-derived mesenchymal stem cells transfected with the BDNF gene can differentiate into nerve-like cells in vitro, which may enable the generation of sufficient quantities of nerve-like cells for treatment of neuronal diseases.

[Effect of brain-derived neurotrophic factor gene transfected bone-marrow mesenchymal stem cells on damaged cochlear spiral ganglion cells of guinea pigs].

OBJECTIVE: To investigate the protective role of brain-derived neurotrophic factor (BDNF) gene transfected bone-marrow mesenchymal stem cells (BMSC) on cochlear spiral ganglion cells (SGC) impaired by aminoglycoside antibiotics (AmAn). METHODS: The differentiation of BMSC transfected by BDNF gene (BDNF-BMSC) were detected with immunohistochemical examination of Nestin, neuron-specific enolase (NSE), and glial fibrillary acid protein (GFAP) antibody in vitro. BDNF gene transfected BMSC were transplanted into the cochleae of guinea pigs deafened by amikacin, while the control groups were designed in which artificial perilymphatic fluid (APF), BMSC or BDNF gene was injected into cochleae alone. The cochleae were obtained on the week 1, 2 and 4 after injection, respectively, paraffin-embedded, and cut in a paramodiolar plane subsequently. The histopathological changes of cochleae were observed, the density of SGC was calculated by staining with HE, and the corresponding optical density (COD) was calculated with immunohistochemical staining using NSE antibody. And the protective role of various groups on the cochlear SGC were compared. RESULTS: The positive staining rate of BDNF gene transfected BMSC with Nestin, NSE and GFAP antibody were all higher than that of BMSC in vitro (P < 0.01). After transplantation into cochleae, the differences of SGC density and COD among various groups were all significant on the same time points (P < 0.05). The SGC density and COD of the BDNF gene transfected BMSC group were the highest. The SGC density and COD of various groups on week 4 were all obviously decreased than those on week 1 and 2 (P < 0.05). CONCLUSION: AmAn-induced SGC damage could be depressed by BMSC, BDNF gene or BDNF gene transfected BMSC transplantation into cochleae, while BDNF gene transfected BMSC showed the best protective role.

[Expression of brain-derived neurotrophic factor modified bone marrow mesenchymal stem cells in the cochlea of drug deafened guinea pigs and its protection role].

OBJECTIVE: To study the expression of brain-derived neurotrophic factor (BDNF) gene modified bone marrow mesenchymal stem cells (MSC) in the cochlea of drug-deafened guinea pigs and its protection to spiral ganglion cells (SGC). METHODS: Guinea pigs deafened by subcutaneous injection of amikacin were randomly divided into two groups, BDNF gene modified bone marrow MSC were injected into the cochlea through fenestration of scala tympani in the experimental group, while artificial perilymphatic fluid were injected in the control group. Experimental animals were executed at 7 and 28 days post-operation. Expression of BDNF mRNA was examined by quantitate real time RT-PCR, histological images of cochlear sections were analyzed to calculate the cellular density of the SGC, and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) was used to identify the apoptotic neurons. RESULTS: The BDNF expressive level in experimental group was higher than in the control group at 7 d and 28 d post-operation, whose differences were both statistically significant (P < 0.01). And, It showed a higher abundance of ganglion cell numbers, as well as a decreased apoptotic index in experimental group compared with the control group at 7 d and 28 d post-operation, whose differences were all statistically significant (P < 0.01). CONCLUSION: BDNF gene modified MSC could maintain expression for at least 28 days after transplantation into cochlea of drug deafened guinea pigs, and protect SGC.

[The incidence of facial nerve dehiscence at mastoidectomy and its risk factors].

OBJECTIVE: To study the incidence and locations of facial nerve dehiscence (FND) in mastoidectomy for the patients with cholesteatoma and chronic otitis media, and to determine its relevance as pre-operative prediction. METHOD: Three hundred and fifteen ears (217 ears with cholesteatoma and 98 with chronic otitis media) undergoing mastoidectomy with or without tympanoplasties were selected for retrospective study, in which the incidence and locations of FND was studied, and the relevance for FND were analyzed by univariate test following by multivariate stepwise logistic regression. RESULT: The presence of FND was 22.9% of total surgical procedures and the locations of FND were 93.1% in the tympanic segment, which was significantly higher than in the mastoid segment. The factors as otogenic facial paralysis, pathologic style (cholesteatoma or chronic otitis media) and lateral semicircular canal (LSC) fistula were related to FND, while others factors as sex, age, revision operations, preoperative complications, dural exposure, sigmoid sinus exposure were not risk factors for FND. CONCLUSION: The incidence of FND was 22.9% in this study, the most common location for FND was in the tympanic segment, therefore, the facial nerves should be especially taken care in mastoidectomy for patients with presence of otogenic facial paralysis, cholesteatoma and LSC fistula.