Objectives To investigate glucocorticoid receptor (GR) and histone deacetylase 2 (HDAC2) gene expression and protein levels in peripheral blood mononuclear cells (PBMCs) of patients with severe or profound sudden sensorineural hearing loss (SSNHL) and to explore the roles of GRs and HDAC2 in glucocorticoid (GC) insensitivity. Methods Fifty-five severe or profound SSNHL patients were enrolled in the study. According to hearing improvement after GC treatment, patients were assigned into two groups: GC-sensitive and GC-resistant. A normal reference group included 20 healthy volunteers without hearing loss. Quantitative real-time polymerase chain reaction and Western blot analyses were used to detect the relative expression of GRα, GRβ, and HDAC2 in PBMCs at the mRNA and protein levels. Results The protein levels of GRs and HDAC2 in PBMCs of SSNHL patients were lower than the normal reference values before GC treatment. Compared with the GC-resistant group, both the mRNA and protein levels of GRα and HDAC2 were significantly increased in the GC-sensitive group after GC treatment. Conclusion A lack of GRα and HDAC2 induction following steroid treatment in GC-resistant SSNHL patients may play a fundamental mechanistic role in GC insensitivity. Response of GRα and HDAC2 to steroid treatment may, thus, predict the prognosis of hearing improvement in SSNHL patients.

目的 探讨绝经后女性良性阵发性位置性眩晕(benign paroxysmal positional vertigo,BPPV)患者BPPV的发生和复发与骨密度、血钙及血清维生素D浓度的相关性.方法 选取2014年1月至2016年12月就诊于东南大学附属江阴医院耳鼻喉科门诊且明确诊断为原发性BPPV的78例(78耳)绝经后女性患者为试验组,根据随访结果 分为BPPV未复发组(63例)和BPPV复发组(15例),选取同期来院体检的自然绝经1年的正常女性30例为对照组.采用双能射线吸收法检测所有受试者腰椎骨密度,抽取清晨空腹外周静脉血4 ml,分别运用半自动分析仪和免疫化学发光法检测血钙浓度和血清维生素D水平.比较各组间骨密度、血钙浓度和血清维生素D水平.结果试验组骨密度下降者占82.15％(64/78),高于对照组(23.33％,7/30),差异有统计学意义(P<0.001).BPPV复发组骨密度T值最低(-1.77±0.63),其次为BPPV未复发组(-1.29±0.82),对照组为-0.63±0.92,组间差异有统计学意义(P<0.001).三组间的血钙浓度差异无统计学意义(P=0.71).对照组的血清维生素D水平(23.98±2.12 ng/ml)高于BPPV未复发组(18.45±4.28 ng/ml)和BPPV复发组(17.15±6.04)ng/ml,差异有统计学意义(P<0.001),BPPV未复发组与复发组之间差异无统计学意义(P>0.05).结论 绝经后女性BP-PV患者BPPV的发生可能与骨密度及血清维生素D水平下降有关,而与血钙浓度可能无关;绝经后女性患者BP-PV的复发则仅与骨密度下降有关.

OBJECTIVE:The relationship between vitamin D and allergic rhinitis (AR) remains unclear. The present study investigated their association by examining serum 25-hydroxyvitamin D (25(OH)D) levels, blood eosinophils, and the expression of vitamin D receptors (VDR) on nasal mucosa in patients with AR.METHODS:A total of 32 patients with persistent AR and 25 controls were enrolled in this study. Serum 25(OH)D levels were detected by enzyme-linked immunosorbent assay, and eosinophils in the peripheral blood were examined by an automated hematology system, while VDR expression on inferior turbinate mucosa was assessed by immunohistochemistry. Furthermore, the correlation of serum 25(OH)D levels with blood eosinophils in persistent AR was analyzed.RESULTS:No significant difference in serum 25(OH)D levels was detected between the AR and control groups (p = 0.371). Interestingly, the serum 25(OH)D levels of the AR group were negatively correlated with blood eosinophil count and its proportion (p = 0.019 and p = 0.010, respectively) even when adjusting confounding factors including age, sex, body mass index, and the season of blood sampling. On the other hand, no significant difference in the expression levels of VDR on nasal mucosa was found between the AR group and the control group (p = 0.231).CONCLUSION:These results suggest that the serum 25(OH)D might be inversely associated with blood eosinophils in patients with persistent AR. However, the relationship between vitamin D and AR still requires further clarification.

近年来,越来越多的研究聚焦于鼓室内灌注激素治疗突发性聋(sudden deafness,SD).2010年1月～2013年3月我科对99例重度以上SD患者分别给予全身或鼓室内注射激素治疗,现将治疗结果报道如下. 1资料与方法 1.1 一般资料.2010年1月～2013年3月我科收治的SD患者99例;男52例,女47例;年龄32～68岁,平均47岁;均为单耳发病;重度聋73例,极度聋26例;声导抗和头颅CT均显示正常;发病到治疗时间间隔10d以内.用0.5、1.0、2.0 kHz的平均听阈(pure tone average,PTA)评估听力损失程度.

Objective The goal of the present study was to determine the effects of different location perfora-tions on the middle ear transfer functions by measuring the velocity and pattern of the tympanic membrane vibration in guinea pigs with intact and perforated TM .Methods A laser Doppler vibrometer was used to measure the sound transmission properties on the movement of the tympanic membrane on 6 fresh temporal bones of guinea pigs in the frequency range of 0 .5~8 kHz at 90 dB SPL .The velocity was measured at three points on the manubrium before and after a different location perforation was made with a 20-gauge needle(1 .1 mm diameter) .The locations of the perforations were in anterior -inferior quadrants of left ears and in posterior -inferior quadrants of right ears .And the velocity was measured from 6 poins with the intact tympanic membrane .Results The highest velocities were re-corded at the site of inferior umbo throughout the frequency range .The manubrium vibration velocity losses were noted in the perforated ears below 1 .5 kHz ,the velocity losses were smaller above 1 .5 kHz ,and the maximum ve-locity loss was about 7 dB at 500 Hz with the posterior -inferior quadrant perforation .Although there was a trend for anterior-inferior perforations to show a slighter loss than posterior -inferior perforations in velocity below 1 .5 kHz ,no statistical differences in velocity loss were found between different perforations .The average ratio of short process velocity to umbo velocity at all frequencies was approximately 0 .5 ,and the ratios were found no systematic differences before and after perforation at almost all frequencies from different perforations .Conclusion The manu-brium vibration velocity losses from TM perforation are frequency -dependent and the largest losses occur at the lowest sound frequencies .Different sites of small perforation have not any important effects on middle ear sound transmission .

Background: Glucocorticoids (GCs) are widely used to treat sudden sensorineural hearing loss (SSNHL) and significantly improve hearing. However, GC insensitivity has been observed in some patients of SSNHL.Objective: To study the correlation between GR expression in peripheral blood mononuclear cells (PBMCs) and in the cochlea of guinea pigs at mRNA and protein levels.Methods: One group of guinea pigs received dexamethasone (10 mg/kg/day) intraperitoneally for 7 consecutive days (dexamethasone group), and another group of guinea pigs received normal saline (control group). Real time PCR and Western blotting were used to detect the expression of GR mRNA and GR protein in PBMCs and the cochleae.Results: The GR mRNA and GR protein were detected in both PBMCs and the cochlear tissue of guinea pigs. GR mRNA and GR protein levels in PBMCs were positively correlated with those in the cochlea. The expression of GR mRNA and GR protein was significantly increased in the dexamethasone group compared to the control group.Conclusions: Levels of GR mRNA and GR protein in the PBMCs were positively correlated with those in the cochlea of guinea pigs. Systemic dexamethasone treatment can significantly up-regulate GR expression in PBMCs and in the cochlea. Measurement of the GR level in PBMCs could be used as an indicator of GR level in the cochlea.

Objective To explore the correlations between the GR isoforms exprossion level in PBMCs and the prognosis in patients with SSNHL and to study the possible regulating effects of GC treatments on the GR isoforms expression level. Methods Fasting blood samples were harvested from 27 patients with SSNHL.PBMCs were isolated by Ficoll density gradient,and the amount of GRα mRNA,GRβ mRNA and SRp30c mRNA were determined by the quantitative real time PCR.The patients were followed up for more than 3 months to evaluate their hearing improvements.The patients were classified into GC-sensitive group(15 cases,PTA improvements ≥15 dB) and GC-resistant group(12 cases,PTA improvements <15 dB).And 6 volunteers with deviation of nasal septum were enrolled as the control group.The statistical difference of the differential mRNA expression of GRα mRNA,GR β mRNA,SRp30c mRNA in PBMCs and the ratio of GR α mRNA/ GR β mRNA were compared among the three groups,before and after intratympanic GC perfusion by SPSS 18.0 software. Results A total of 27 patients with SSNHL were enrolled in this study.20 patients received intratympanic GC perfusion and 7 patients received GC treatments in vein.After treatments,15 patients were GC sensitive and 12 patients were GC resistant according to their hearing improvements.The expression of GRα mRNA was significantly higher than GRβ mRNA,which was significantly associated with the expression of SRp30c mRNA in all the three groups.The expression of GRα mRNA,GRβ mRNA,SRp30c mRNA and the ratio of GRα mRNA/GRβ mRNA before treatments had no statistical differences among the three groups.The level of GRα mRNA and GRβ mRNA was down-regulated by intratympanic GC perfusion in GC-sensitive group,but not so in GC-resistant group. Conclusion The different expression level of GRα mRNA,GRβ mRNA,SRp30c mRNA and the ratio of GRα mRNA/GRβ mRNA in PBMCs before treatment were not associate with the hearing outcome in this study,and they shouled not be used to predict the hearing recovery in SSNHL patients.But the down-regulated expression of GRα mRNA and GRβ mRNA after intratymanic GC perfusion may indicate a better hearing recovery.Further studies are needed.

Objective To study correlation of glucocorticoid receptor (GR) contents between peripheral blood mononuclear cells (PBMC) and the cochlea in guinea pigs by measuring the expression of GR mRNA and GR protein and the effects of systemic injection of dexamethasone on the expression level. Methods Healthy guinea pigs (n=32) were randomly divided into two groups (dexamethasone treatment and control groups). Dexamethasone (5 mg/ml, 10 mg/kg/day) was injected intraperitoneally in the 15 guinea pigs of the dexamethasone group for 7 continuous days, while same volume physiological saline was injected intraperitoneally in the 17 guinea pigs of the control group. PBMCs and bilateral cochlear tissue were collected. Real time PCR and Western blot were used to detect the expression level of GR mRNA and GR protein in PBMCs and cochlear tissues.The data were analyzed with the SPSS 16.0 software. Results GR mRNA and GR protein were expressed in both PBMCs and cochlear tissues. The expression levels of GR mRNA and GR protein increased significantly in the dexamethasone treatment group and showed close correlation between PBMCs and the cochlea . Conclusions The levels of GR mRNA and GR protein in the PBMC indirectly reflect those in the cochlea in guinea pigs.

Background: It has been believed that location of the perforation has a significant impact on hearing loss. However, recent studies have demonstrated that the perforation sites had no impact on hearing loss. We measured the velocity and pattern of the manubrium vibration in guinea pigs with intact and perforated eardrum using a laser Doppler vibrometer in order to determine the effects of different location perforations on the middle ear transfer functions.Methods: Two bullas from 2 guinea pigs were used to determine stability of the umbo velocities, and 12 bullas from six guinea pigs to determine the effects of different location perforations on sound transmission. The manubrium velocity was measured at three points on the manubrium in the frequencies of 0.5-8 kHz before and after a perforation was made. The sites of perforations were in anterior-inferior (AI) quadrants of left ears and posterior-inferior (PI) quadrants of right ears.Results: The manubrium vibration velocity losses were noticed in the perforated ears only below 1.5 kHz. The maximum velocity loss was about 7 dB at 500 Hz with the PI perforation. No significant difference in the velocity loss was found between AI and PI perforations. The average ratio of short process velocity to the umbo velocity was approximately 0.5 at all frequencies. No significant differences were found before and after perforation at all frequencies (p>0.05) except 7 kHz (p = 0.004) for both AI and PI perforations.Conclusions: The manubrium vibration velocity losses from eardrum perforation were frequency-dependent and the largest losses occur at low frequencies. Manubrium velocity losses caused by small acute inferior perforations in guinea pigs have no significant impact on middle ear sound transmission at any frequency tested. The manubrium vibration axis may be perpendicular to the manubrium below 8 kHz in guinea pigs.

Laminated images of microstruetures of biological tissues have been obtained by the scanning electron-acoustic thermal wave microscopy by suitably adjusting the phase,because the acoustic signals produced by the thermal sources at different depths are with different phases. On the other hand, in order to theoretically simulate the laminated imaging by the electron-acoustic microscopy,thermal conduction and thermo-elastic equations are introduced to obtain the distributions of the temperature field and then the acoustic field in the samples. Based on the solutions of the equations and related boundary conditions,the depths of laminated images can be theoretically estimated quantitatively as the thermal diffusivities of the samples are determined.Meanwhile,the thermal diffusivity of the samples can also be measured by the electron-acoustic detection technique. For the biological tissues(heart and liver of rats)the thermal diffusivities are measured,which are in the range of 0.45-0.65 mm2/s,and the measured depths are about several tens micrometers.Therefore,the depth profiling of the biological tissues can be evaluated quantitatively by the theoretical fitting to the experimental laminated images.

Objective To characterize the mechanism of the middle ear sound transmission by measuring the velocity of the umbo in guinea pigs.Methods A laser Doppler vibrometer was used to measure the sound transmission properties on the movement of the umbo on 18 fresh temporal bones of guinea pigs at the frequencies range of 0.5～8 kHz at 90 dB SPL.Results Repeated measurements taken at the umbo showed that the frequency response maintained a similar shape.The velocities did not exhibit large variations with the changes within 1 dB below 3 kHz and the changes in the range of 7 dB above 3 kHz.The frequency responses measured at the normal umbo in guinea pigs exhibited two peaks at the range of 0.5～8 kHz.Conclusion Results indicated that the frequency responses measured at the normal umbo in guinea pigs exhibited a peak at 2kHz and a second peak at 4.5 kHz.The measured velocity and vibration responses in the left and right ears of the normal umbo in guinea pigs were correlated.

Objective To explore the role of transforming growth factor β1(TGF-β1) and transforming growth factor-β2(TGF-β2) in the onset and development of otitis media with effusion (OME) in the guinea pig model.Methods The expression of transforming growth factor-β1 and transforming growth factor-β2 in the tympanum mucosa of middle ear and venous serum in guinea pigs with OME were examined by means of ELISA and Western blot.Results Compared with control group,expressions of TGF-β1 in the tympanum mucosa of middle ear and venous serum in test group were indistinctive at 1 d and 3 d,more intensive at 7 d,and reached maximum at 30 d.There was significant statistic difference among groups(P<0.001).Compared with control group,expressions of TGF-β2 in the tympanum mucosa of middle ear and venous serum in test group increased from 1 d,reached maximum at 14 d,then descended.There was significant difference among groups (P<0.001).Conclusion TGF-β1,TGF-β2 were expressed differently in the tympanum mucosa of middle ear and venous serum in the middle ear mucosa of the guinea pigs with OME at different stages.This indicates that TGF-β1,TGF-β2 may play a diverse and key role in the onset and development of OME.

糖皮质激素(glucocorticoid,GC)已广泛应用于自身免疫性内耳病、突发性聋、梅尼埃病等内耳疾病的治疗~([1,2]).在细胞水平GC与其受体结合,激活的受体转移至细胞核内调节基因的表达从而在靶器官发挥生物学效应.组织对GC的反应主要取决于组织中糖皮质激素受体(glucocorticoid receptor,GR)的含量~([3]).目前GR在内耳的作用机制及患者对GC敏感性不同的原因尚不十分明确,本文对GR的分子生物学特征以及GR在内耳作用的研究进展进行综述.