We evaluated the histologic and histochemical (succinate dehydrogenase, SDH) changes in cochleas of guinea pigs after non-focused ultrasound (NFU) irradiation. For this purpose, the cochleas of 50 guinea pigs (both ears in Groups 1-10) were irradiated at 2.5-8.0 MHz NFU for 6 h and the histologic/histochemical changes were determined. Our data show that after NFU irradiation for 6 h, no histological changes were observed in the cochleas by silver staining and scanning electron microscopic examination; however, the SDH activity of the basilar membranes and stria vascularises at the corresponding position decreased significantly. The SDH values of basilar membranes and stria vascularis were higher at 8 h than at 30 min after the NFU irradiation. SDH activity of the outer hair cells presented more declination than that of the inner hair cells. It was, therefore, concluded that a certain dosage of NFU irradiation at various frequencies could lead to metabolic changes in the basilar membrane and stria vascularis at different areas of cochlea. Moreover, these changes were found to be reversible or partially reversible. These changes also suggest that the cochlear hair cells located at different areas might be related to ultrasonic perception.

目的探讨内耳免疫反应过程TNF-α/TNFR1的表达及作用。方法选用雌性白色豚鼠45只,随机分为实验组29只,对照组16只,以钥孔虫戚血蓝蛋白（keyhole limpet hemocyanin,KLH）全身免疫后,实验组以相同抗原进行内耳免疫,对照组内耳注射等量的磷酸盐缓冲生理盐水（phosphate buffered saline,PBS）,分别在内耳免疫术后1、3、5、7和14 d处死动物,取内耳免疫侧耳蜗做石蜡切片。通过免疫组化检测内耳TNF-α的表达,原位杂交检测内耳TNFR1mRNA的表达。结果对照组TNF-α蛋白表达阴性。实验组除内耳免疫第1天Corti器、侧壁和螺旋神经节与内耳免疫第3天侧壁外,其余切片都有TNF-α蛋白阳性表达,免疫组织化学染色平均吸光度值均显著高于对照组（均P＜0.01）。对照组TNFR1 mRNA表达阴性。实验组除内耳免疫第1、3、5天Corti器外,其余切片均有TNFR1 mRNA阳性表达,TNFR1 mRNA原位杂交染色平均吸光度值均显著高于对照组（均P＜0.01）。结论 TNF-α/TNFR1是调节内耳免疫反应的重要细胞因子和信号转导途径之一。

OBJECTIVE To analyses the clinical characteristics and electrophysiological finding of 106 patients with auditory neuropathy (AN). Investigate the differential curve type of pure tone audiogram and the abnormal ABR. METHODS Review the history of patients, pure tone audiometry, middle ear acoustic reflexes, auditory brainstem response, distortion product otoacoustic emission and radiologic imaging studies of the brain of 106 patients with AN during December 2001 to May 2007 in retrospect. RESULTS The 106 patients with AN were of both genders. The age were between 11-37 years old, and the average age was 17.5 years old. The most patients were adolescence (70.8%). Twelve cases of the 106 patients had evidence of other peripheral neuropathy in addition to hearing loss. Another 94 patients were idiopathic origins. The pure tone audiogram showed a minimal to moderate sensorineural hearing loss at low frequencies 0.5 kHz and 0.25 kHz in 209 ears (98.6%). The average hearing threshold (WHO 1997) in 23.2% of disordered ears at less than 25 dB in the "normal" range. Auditory brainstem response could not be recorded in 124 ears (58.5%) at maximum stimulus. The other 88 ears showed 1 or 2 wave, but the wave were small. There were 23 patients which one side ear ABR was 1 or 2 small waves presented, but the contralateral side were all waves absent. In 3 cases of AN with other peripheral neuropathy which ABR were both ears 1 or 2 small wave ear recorded. However, 1 patient in our sample could not be detected distortion product otoacoustic emission at 3-6 kHz (left ear) and 5-6 kHz (right ear). CONCLUSIONS AN was not rare in adolescence. The average hearing threshold for AN should be discussed.

外淋巴瘘(perilymphatic fistula)是指充满液体的内耳与含气的中耳和乳突腔之间的异常交通,其中圆窗膜和(或)环状韧带撕裂又称迷路(内耳)窗膜破裂.

OBJECTIVETo investigate the character of 80 Hz auditory steady-state evoked potentials in auditory neuropathy (AN) patients and the correlation between it and ABR.METHODThe 80 Hz auditory steady-state evoked potentials and ABR to 95 dBnHL (1000 Hz) tone-pip stimuli were recorded from the scalp in patients with AN and normal subjects. The difference of the waveforms between AN ears and normal ears was compared.RESULTIn 85.4% AN ears the 80 Hz auditory steady-state evoked potentials and ABR were simultaneously present. Their amplitudes were low or extremely low. The I and III waves of ABR were absent. The 80 Hz auditory steady-state evoked potentials and ABR were simultaneously absent, in 14.6% AN ears, but were present in all normal ears.CONCLUSIONThe 80 Hz auditory steady-state evoked potentials and the V wave of ABR may come from the same brainstem sources.

OBJECTIVE:To investigate the auditory characteristics of guinea pigs immunized with purified P0 protein from inner ear of guinea pigs. METHOD:Purified inner ear P0 protein was made by Preparative SDS-PAGE. To evaluate the results of Auditory Brainstem Response (ABR), Compound Action Potention (CAP), and Distortion Product Oto-acoustic Emissions (DPOAE) in guinea pigs immunized with purified guinea pigs inner ear myelin protein P0. RESULT:Seven ears of the guinea pigs immunized with purified inner ear P0 protein developed hearing loss. In the ABR study, peak latencies of wave I, III and the interpeak latency of I -III, I-IV were elevated in this group of guinea pigs compared with the control group (P < 0.01). But the interpeak latency of III-IV did not change. In the CAP study, the threshold elevated and latency prolonged (P < 0.01). No significant change of DPOAE was found in the P0-sensitized guinea pigs when compared to controls (P > 0.05). Even though the contra-lateral suppressive effect had the depressed tendency, but there was no significant different when compared with the controls (P > 0.05). CONCLUSION:Purified inner ear P0 protein is an important autoimmune inner ear antigen and can develop autoimmune disease of the auditory nerve.

近年来,随着耳外科技术的发展及生物医学工程领域的进步,围绕着感音神经性耳聋、耳发育畸形、复杂中耳病变等的替代治疗,取得了很大的进步,植入式助听装置为重要代表之一.

OBJECTIVE To study the enzyme changes of stria vascularis of the cochlea in guinea pigs after ultrasound irradiation. METHODS After 30 minutes or 8 hours of 2.5 mHz,8.0mHz ultrasound irradiating the cochlea of guinea pigs(45 ears)for 6 hours,the SDH(succinate dehydrogenase) activity of the stria vascularis was observed. RESULTS The ultrasound irradiation for 6 hours had led to enervation of the activity of SDH of the stria vascularis in the (0.91±0.22)mm ranges of the second turn or (0.97±0.26)mm ranges of the first turn,respectively. The A value of SDH of the stria vascularis was higher in the group of 8 hours than the group of 30 minutes after ultrasound irradiation. CONCLUSION Ultrasound(2.5mHz、 8.0mHz )irradiation the cochlea for 6 hours can cause pathological changes in the stria vascularis of different areas. Moreover,the pathological changes were restorable or part restorable in some amount after ultrasound irradiation.

OBJECTIVE:To investigate the long-term outcome and clinical value of modified radical mastoidectomy with mastoid obliteration using pedicled combined flap of postauricular musculo-periosteal and ear canal skin flap in conjunction with bone paté.METHODS:During 2 years from April 2003 to March 2005, 71 otitis media patients (71 ears) with cholesteatoma were subjected to this kind of operation.RESULTS:The follow-up period was more than 2 years. The period of complete reepithelialization ranged from 3 weeks to 1.5 months, with the mean period of 29 days. All of the patients, treated by the described method of operation had a dry, disease-free mastoid of ear.CONCLUSIONS:Mastoid obliteration with pedicled combined flap of postauricular musculo-periosteal and ear canal skin flap in conjunction with bone paté, had the advantages as follows: (1) Healing of the mastoid cavity in a short time. (2) Better reepithelialization of the obliterated mastoid cavity. (3) No need of skin grafting in the mastoid cavity. (4) High rate of the dry ear in postoperation. (5) nearly no need of the mastoid cavity cleaning postoperatively.

OBJECTIVETo purify P0 protein from guinea pig's inner ear by preparative SDS-PAGE and to study the possible role it may play in the etiology of autoimmune inner ear disease.METHODA mixture of membraneous proteins of inner ear was separated by preparative SDS-PAGE. The corresponding band at 30,000 was cut and electrically eluted. The protein collected was identified by analytical SDS-PAGE and Western blot assay. A group of 20 guinea pigs were immunized with P0 protein emulsified in complete Freunds adjuvant, another 10 guinea pigs were immunized with complete Freunds adjuvant only as control. The guinea pigs' hearing thresholds, serum IgG level and morphological changes in the inner ear were investigated. The distribution of P0 protein in the cochlear was detected by immunohistochemical technique.RESULTThe purity of the protein was demonstrated by a single band at the 30000 site in SDS-PAGE, which was identified as P0 protein by western blot analysis assay. About 17.5% P0-immunized guinea pigs showed increased hearing thresholds, elevated IgG level (F = 6.48, P<0.01), as well as a decreased number of spiral ganglion cells and inflammatory cell infiltration in the cochlear nerve region. The P0 protein is distributed in the cochlear nerve and spiral ganglion only.CONCLUSIONP0 protein from guinea pigs in ner ear can be successfully purified by preparative SDS-PAGE and an animal model of experimental autoimmune inner ear disease induced by P0 protein was successfully established.

Objective To investigate the clinical characters of congentital aural atresia and the surgical treatment of the disease. Methods In this series, 64 ears with congentital aural atresia operated on in our hospital during the period from January 1999 to December 2003 were restrospectively analysed. Results Forty-one of these ears were surgically corrected by anterior approach, the remaining 23 were corrected by transmastoid approach. Of all cases, the tympanic cavity was encountered without difficulty in 61 ears except for 3 ears. Ossicular chain anomalies occurred in all these 61 ears, and varying degrees of abnormal facial nerve were found in 40 ears ( 65.57% ). Postoperative follow-up ranged from 1 to 5 years. The hearing improvement was observed in 49 ears( 80.33% ), of which 32 ears had hearing improvement greater than 30 dB and the air-bone conduction gap was within 15 dB( 52.46% ). The external auditory stenosis occurred in 6 of these ears during the priod from postoperative 3 months to 2 years, of which 4 ears obtained goood results after the second operation. Lateralization of the tympanic membrane presented in 2 ears and infections of the cavity in other 2 ears. Facial paralysis occurred postoperatively in 3 ears, which recovered 2 to 6 months later. Conclusions The surgical procedures had no regular pattern, depending upon what one sees during the operation. The sclerotic and diploetic atretic bone could be indications for superoanterior surgical approach, whereas it would be wiser for the mixed and pneumatic mastoid to be corrected by transmastoid approach.

OBJECTIVE:To further evaluate the therapeutic effect of hyperbaric oxygen (HBO) to idiopathic sudden deafness. METHOD:A total number of 236 medical records of inpatients with idiopathic sudden deafness during 1997-2004 were collected from the Wuhan Union Hospital. Single factor analysis was done to evaluate the therapeutic effect with ten factors with SPSS analysis software. We divided the cases into HBO group and non-HBO group. The logistic regression was used in multivariate analysis. RESULT:Single factor analysis indicated that sex, ear side, presence of tinnitus, inducement and period of treatment had no effect on the prognosis, while age, course, presence of vertigo, a sensorineural hearing loss history with the other ear, degree of deafness had some effect on the prognosis. Multivariate analysis indicated: the effective (hearing threshold has elevated above 15 dB) ratio of HBO group comparing to the control group (non-HBO group), the difference was significant (P<0.01). CONCLUSION:The first visit after 14 days, the presence of vertigo, a sensorineural hearing loss history with the other ear and complete deafness may imply poor prognosis. HBO is effective for sudden deafness.

OBJECTIVE:To study the cell proliferation in endolymphatic sac(ES) during the secondary immune response in inner ear.METHODS:Fifteen healthy, female SD rats were employed in the experiment. Sensitized systematically with keyhole limpet hemocyanin (KLH), the animals were inoculated with the same antigen through cochlea basal turn into the labyrinth. Administrated 5-bromo-2'-deoxyuridine (BrdUrd) intraperitoneally, the rats were sacrificed and the temporal bones were harvested at 3, 7, 14 day after labyrinth vaccination respectively. The frozen sections of the decalcified samples were dealt with H-E staining and immunohistochemical methods to investigate the cellular infiltration, BrdUrd and IgG positive cells in the ES.RESULTS:While the ES lumen and perisaccular region were infiltrated with mononuclear-phagocyte at the 3rd day post-vaccination, plasmocyte and lymphatic cells become the predominant infiltrating cells in the ES at the 7th day. The KLH in the ES lumen were phagocytized at the 3-7th day post-vaccination. S-phase cells and IgG positive cells in ES increased markedly in the 3rd day and 7-14 days post-vaccination respectively with similar localization.CONCLUSION:The activity of cell proliferation in the ES enhance and local proliferated cells may differentiate in situ into immunocompetent cells during the secondary immune response against TD antigen in the inner ear. This indicates that ES plays an important role in immune response of inner ear.

OBJECTIVE:To study the genetic background, clinical features, diagnosis, management, and prenatal testing and counseling of Treacher Collins syndrome (TCS).METHOD:The clinical data of a patient and his mother suffering from TCS was reported and the relevant literatures were also reviewed.RESULT:TCS is an autosomal dominant disorder characterized by craniofacial mal-development, including antimongoloid slant of the eyes, micrognathia, microtia and other deformity of the ears. TCS results from the mutation in TCOF1 gene (Treacher Collins-Franceschetti Syndrome gene), located in 5q32-q33.1. Diagnosis of TCS is made through clinical evaluation, radiographic examination, and molecular genetic analysis. Treatment is tailored to the specific needs of each individual by a multidisciplinary craniofacial management team. The choosing of optimal operation time and method is very important. Prenatal testing and genetic counseling for risky pregnancy is necessary.CONCLUSION:The stepwise regimen of treatment is craniofacial reconstruction, orthognathic procedures, orthodontic alignment of the teeth, nasal reconstruction, external ear reconstruction and external auditory canal and middle ear reconstruction.

OBJECTIVE:To identify the clinical and anatomical presentations and to discuss the guidelines for surgical management of anomalies of the first branchial cleft.METHOD:Twenty-one patients with first branchial cleft anomalies were treated in our department between January 1994 and December 2004, their clinical data were retrospectively analysed.RESULT:Surgery was performed on all patients. Among them 13 were males and 8 females, ranging in age from 1.5 to 33 years with an average of 15 years. Anatomically, 3 types of first branchial cleft anomalies were identified: fistulas (n = 17), cysts (n = 2), and fistula combined with cyst (n = 2). Before definitive surgery, soma patients (n = 4) underwent incision and drainage for infection owing to the difficulties in diagnosing this anomaly. Methylthioninium Chloride was used in almost all cases for tracking the fistulous during operation. Wide exposure is necessary in many cases,and a standard parotidectomy incision allows adequate exposure of the anomaly and preservation of the facial nerve.CONCLUSION:Complete removal without complications depends on a good understanding of regional embryogenesis, an awareness of the different anatomical presentations, and a readiness to identify and protect the facial nerve during resection.

Objective To investigate whether apoptosis is invovled in the pathogenesis of the immune response of the inner ear,and to explore the relation between signal transportation of Fas,FasL and apoptosis.Methods Sixteen healthy,female guinea pigs were employed in the experiment.Sensitized systematically with keyhole limpet hemocyanin(KLH),the KLH-immunized animals were inoculated with the same antigen,while the control animals were injected PBS through cochlea basal turn.The animals were sacrificed 5 days after inner ear vaccination.Paraffin sections of cochleas were stained by TUNEL assay to identify inner ear cells undergoing apoptosis,and immunohistochemistry was used to detect the expression of Fas and FasL in the inner ears.Results TUNEL-positive cells were found in the KLH-immunized inner ears but not in the control inner ears except a few positive cells in the supporting cells,the stria vascularis cells and the spiral ganglion cells.The positive cells were the hair cells in the Corti's organ,the marginal cells in the stria vascularis and the neurons in the spiral ganglion.High expression of Fas and FasL could be detected in the Corti's organ,the stria vascularis,the spiral ligament and the neurons of the spiral ganglion in the KLH-immunized inner ears.Low expression of Fas could be detected in the stria vascularis and the neurons of the spiral ganglion in the control inner ears,but no positive FasL cells were found in the control inner ears.Conclusion These findings suggest that apoptosis is involved in the pathogenesis of the immune response of the inner ear and the Fas-FasL is one of the important pathways of signal transportation.

OBJECTIVE:To compare the pharmacokinetics of Methylprednisolone, Dexamethasone and Hydrocortisone in the inner era of guinea pig after administered via round window.METHOD:Guinea pig was divided into groups chronologically. After a drop of 100 g/L corticosteroid applied into the round window niche, the tympanic perilymph was sampled chronologically. Concentration of the three corticosteroid in perilymph were measured via HPLC and analyzed by 3P97 software.RESULT:The highest concentration of the three drug appeared at 30 min, 60 min and 30 min. Then the concentration declined rapidly.CONCLUSION:All three corticosteroid permeate the round window membrane easily, but the concentration declined rapidly after the peak appearing.

OBJECTIVE To investigate whether apoptosis is involved in the pathogenesis of the immune response of inner ear, and relation between signal transportation of Caspase-3 and apoptosis. METHOD Forty-five healthy, female white guinea pigs were employed in this study. Sensitized systematically with keyhole limpet hemocyanin (KLH), the KLH-immunized animals were inoculated with the same antigen, and the control animals were injected PBS through cochlea basal turn. All animals were sacrificed respectively at 1st, 3rd, 5th, 7th and 14th day after inner ear vaccination. The auditory brainstem response (ABR) was measured before the KLH injection to the inner ear and just before sacrificing the animal. Paraffin sections of cochleas from animals were stained using a TUNEL assay to identify inner ear cells undergoing apoptosis,and immunohistochemistry method was used to detect the expression of Caspase-3 in the inner ears. RESULT After immunization, the ABR thresholds of the KLH-challenged ears were significantly elevated by comparing with the preimmunization (P < 0.05), but after injection, there was no significant difference in the ABR thresholds of the PBS-injected ears compared with preinjection. TUNEL-positive cells were found in the KLH immunized inner ears but no TUNEL-positive cells were observed in the control inner ears except for a few positive cells in the supporting cells, the stria vascularis cells and the spiral ganglion cells. The positive cells are the outer hair cells in Cortis organ, and the marginal cells in the stria vascularis and the neurons in the spiral ganglion. And, under morphological analysis by light microscope, these cells have the features characteristic of apoptosis. Apparent Caspase-3 immunoreactivity expression could be detected in Corti's organ, the lateral wall and the neurons of the spiral ganglion in the KLH-immunized inner ears from 5th day after inner ear vaccination, but no cells staining positive for Caspase-3 were found in the control inner ears. CONCLUSION Apoptosis is induced by the immune response of inner ear; Caspase-3 cascade has important effect on the course of apoptosis.

OBJECTIVE: To observe the distribution of CGRP in guinea pig cochlea. METHOD: CGRP was labeled by immunostaning technique. Distribution of CGRP immunoreactivity was observed with light microscopy. RESULT: CGRP immunoreactivity was located in SGN, nerve fibers at osseous spiral lamina, stria vascularis, IHC, adjacent area of OHC and Deiters' cells. CONCLUSION: CGRP distributes extensively in guinea pig cochlea. CGRP may play an important role as neurotransmitter and/or neuromodulator to hearing function by modulating HC, supporting cells, SGN and other neurons along the process of auditory nerve. It may have effect on on cochlear blood flow, too.