Objective:To investigate the generative or regenerative hair cell and studes of molecular and genetic in the inner ear,the long-term culture systems of utricular sensory epithelial cell of the rats were established.Method:Utricular sensory epithelial of postnatal day 1 rats was isolated by mechanical dissociation.The explants were digested by thermolysin,then transfrred to an aliquot containing trypsin and collagenase for incubation to harvest the pure utricular sensory epithelial cell. USEC wes cultured in Dulbecco modified Eagle medium(DMEM) and passaged.USEC of the 25 passages observed by inverted microscope and ultrastrctural examination with transmission electron microscope.Immunocytochemical method with cytokeratin 18 ,vimentin,Brn3.a and Calretinin; reverse transcription PCR with mRNA of AchRa9 and Myosin Ⅶa.The markers of hair cells were used identify the characteriztion of USEC of the 25 passages.Result:USEC have been cultured for more than 6 months and passaged 25 passages.USEC of the 25 passages showed a large,flat,polygonal epichlial morphotype with big,round neuclei.The cells grew into monolayer,cobblestone-likeappearance and showed Dome formation. The cells expressed cytokeratin 18 , not expressed vimentin,has rich microvilli and complex tight junction,which indicated the epithlial origination of USEC.Coexpressed of the hair cell characteristic markers Brn3.a,Calretinin and mRNA of AchRa9,Myosin Ⅶa was identified the culture cell that may represent rat progenitor hair cell.Conclusion:The long-term culture systems of utricular sensory epithelial cell of the rats were successfully established. The long-term culture USEC coexpressed the characteristic markers of the hair cell ,which identified the culture cell may represent progenitor cell of rats hair cell .It may provide valuable cell sources for in-depth investigation the mechanisms of hair cell generation or regeneration and studes of molecular and genetic in the inner ear.

Objective To observe the expression of gamma-amino butyric acid A receptor(GABAA receptor)?3、?1、? subunits in rat flocculus following unilateral labyrinthectomy(UL).Methods 24 wistar rats were randomly divided into two groups.18 animals received unilateral labyrinthectomy while the others maintained labyrinthine well.After removing left labyrinthine,the change of GABAA receptor ?3、?1、? subunits was detected by immunohistochemistry.Results GABAA receptor ?1、? subunits in flocculus was induced on the operated side after unilateral labyrinthectomy.The most expression was on the 1st day flocculus of following UL.The expression is descending from the 3rd day to 7th day flocculus of following UL.The GABAA receptor ?3 subunit immunostaining in flocculus was weak and did not change in the process of vestibular compensation.Conclusion GABAA receptor ?1、? subunits were induced increase in the flocculus after unilateral labyrinthectomy.The alteration in the resting discharge of the central vestibular neurons may be caused by the increase of GABAA receptor ?1、? subunits in the flocculus.But the significance of the change of GABAA receptor ?1、? subunits in the vestibular compensation is still unknown.Our study suggests that GABAA receptor ?3 subunits may not participate in constructing GABAA receptor in the flocculus.

OBJECTIVE To investigate the effect of canalith repositioning procedure(CRP)on the postural stability in patients with benign paroxysmal positional vertigo of the posterior semicircular canal(BPPV-PSC).METHODS 25 patients with BPPV-PSC were taken the posturography before and after the CRP.The posturography test has four conditions including①firm surface with eyes open(EO);②firm surface with eyes closed(EC);③foam surface with EO;④foam surface with EO.The body sway velocity(SV)recorded by the posturography was analyzed as parameter of postural stability.The postural stability of the patients with BPPV-PSC was compared with that of the matched control group.RESULTS ①The SVs of the BPPV-PSC patients before the CRP were greater than those of control group(P

Objective To investigate the possible association of mtDNA deletion in spiral ganglion of cochlear and cochlear nucleus of brain in young and elder rat with presbyacusis.Methods The mtDNA 4834 deletion in spiral ganglion of cochlear and cochlear nucleus of brain in rat was analyed by PCR.Results The deletion rate of mtDNA 4834 in rat with presbyacusis was 66.7% in spiral ganglion, 100% in cochlear nucleus of brain.While no deletion was detected in young rat.Conclusion Deletion in mtDNA of the spiral ganglion of cochlear and cochlear nucleus of brain was found in rat with presbyacusis, and it may play an important role in the pathogens of presbyacusis.

Objective To investigate the cell origination of progenitor cells of the mammalian regenerative hair cell.Methods Utricular sensory epithelial of postnatal day 1 wister rats was isolated by mechanical dissociation.The explants were digested by thermolysin,then were transfrred to an aliquot containing 0.125% trypsin and 0.125% collagenase for incubation to harvest the pure utricular sensory epithelial cell(USEC).USEC were cultured in Dulbecco Modified Eagle Medium(DMEM),and observed daily by inverted microscope.Immunocytochemical staining with cytokeratin 18,vimentin,Brn3a and Calretinin,ultrastrctural examination with transmission electron microscope,and reverse transcription PCR with mRNA of p27kipll,AchRa9 and Myosin Ⅶa.The markers of hair cells and supporting cells were used to identify the origination and characteriztion of USEC.Results USEC showed a large,flat,polygonal epithelial morphotype with big,round neuclei.USEC monolayer showed "cobblestone-like" appearance.Some cells showed "Dome" formation,probably due to fluid collection underneath the cell monolayer.USEC expressed cytokeratin 18 but did not express vimentin,and had rich microvilli and complex tight junction,which indicated the epithelial origination of USEC.USEC expressed Brn 3a,calnevimin and mRNA of p27kipl,AchRa9,myosin Ⅶ.Conclusion USEC coexpressed the characteristic markers of the hair cell and supporting cells.This indicates that the culture cells may represent progenitor cell of hair cell and the progenitor cell may come from supporting cells.

Objective To study the influence factors affecting the blood sodium level of patients with respiratory system diseases for the first time. Methods A retrospective analysis of 557 cases in our hospital from January 1,2013 to April 31, 2013 were analyzed.According to the serum sodium levels, patients were divided into normal blood sodium group and hyponatremia group.The basic data, clinical indexes, prognosis were collected and the relationships between the epidemiology study of hyponatremia and the diseases of respiratory system were analysed.Results 557 cases of respiratory patients, 121 cases were complicated with hyponatremia, the incidence rate was about 21.72%; hyponatremia incidence of the top three diseases of respiratory system:57 cases of pneumonia ( 47.1%) , 26 cases of chronic bronchitis ( 21.5%) , 11 cases of lung cancer ( 9.1%); temperature, diastolic blood pressure, heart rate, WBC, Hb, Alb, BUN, Cl, might had an impact on the level of serum sodium (P<0.05); hyponatremia group prognosis was significantly worse than normal blood sodium group (P<0.05).Conclusion Patients in the department of respiratory internal medicine are easy suffered with hyponatremia, the factors are more, prognosis is poor, requiring clinical attention.

Objective To observe the opening movement of eustachian tube pharyngeal orifice in patients with secretory otitis media(SOM) by cine CT to mearsure the thickness of the lateral ET wall ,and to evaluate the roles played by those peri-tube structure in the pathogenesis of ET obstruction .Methods Twenty -seven SOM patients as experiment group underwent low -radiation dose cine CT scans of the ET ,the image were reconstructed into a cine image to see the opening movement of the eustachian tube pharyngeal orifice .The CT scan of sixty-four non-SOM patient as control group was performed .The images were reconstructed to show the relationship among the in‐ferior turbinate ,the nasopharyngeal soft tissue and the eustachian tube .These serial images were analyzed to meas‐uring the thickness of the lateral wall of ET cartilage segment and to find out whether the gas exist in the pharyngeal recess .Results The opening movement of the pharyngeal orifice was observed in 23 SOM patients ,the mean thick‐ness of the lateral wall of ET cartilage segment in ithe ears studied and healthy ears of SOM patients are 8 .93 ± 1 .6 mm and 8 .89 ± 1 .2 mm ,respectively ,much larger than those of in non-SOM patients ,but with no static signifi‐cance .The pharyngeal recess in 69 .0% (29/42) affected ears of som patients and 58 .3% (7/12) in healthy ears were gas free .This rate in som patients was significantly higher than that in the non-SOM patient group .Conclusion The etiologies of dysfunction of eustachian tube may include hypertrophic inferior turbinate ,chronic inflammation of eustachian tube mucosa ,compression of the nasal pharyngeal soft tissue .The compression of the nasal pharyngeal soft tissue may play an important role in the obstruction of the specific cartilage segment of the ET .

Objective To investigate the optimal b value of diffusion-weighted imaging (DWI)in predicting the grade of cerebral gliomas.Methods 38 patients with pathologically-proved brain gliomas (24 high-grade gliomas and 14 low-grade ones)were studied retrospectively.All patients received conventional MRI and DWI examination with three different b values (1 000 s/mm2 ,2 000 s/mm2 and 3 000 s /mm2 ).The signal features of the tumor on DWI with three different b values were evaluated and compared.The minimum ADC values of the tumors were calculated and compared between each other.Through drawing the ROC curve of different b values, the best diagnostic threshold was found.The sensitivity and specificity in predicting the grade of brain gliomas were assessed using Chi-square test.Results On DWI with b value of 3 000 s/mm2 ,91.6% (22/24)of high-grade tumors showed hyper-intensity, while 85.7%(12/14)of low-grade tumors presented hypo-intensity signals.Regarding hyper-intensity signals as diagnostic criterion for high-grade tumors,the sensitivity and specificity were 91.6% and 100% respectively.When the b value was similar,the minimum ADC value of high-grade glioma was significantly lower than that of low-grade glioma,and statistically significant differences in differentiating high-grade glioma from low-grade glioma existed among three b values (P <0.05).When the ADC 3 000 value<0.74×10 -3 mm2/s was regarded as a standard for the identification of high-grade and low-grade glioma,the sensitivity and specificity were 100% and 87.3%respectively.Conclusion DWI with high b value is more useful than standard moderate b value in preoperative grading the gliomas.When DWI with b value of 3 000 s/mm2 is used,the minimum ADC value will provide quantitative indicators in preoperatively precisely predicting grading glioma.

In this study the relation between the extent and type of disc protrusion and the clinical severity of symptoms of low back pain was observed. Our series consisted of 300 consecutive cases of lumbar disc protrusion,with their diagnosis confirmed by intraoperative observation. Preoperative pain severity was assessed using a back pain scale that combined the symptoms of back pain and sciatica. Intraoperaative pathological findings of disc were documented and comparesd with preoperative symptoms and signs. The pathological changes of disc were classified as following four subtypes: bulging disc (85 cases), protruding disc (104 cases), extruded disc (72 cases) and sequestered disc (39 cases). Our results suggest that severity of pain in patients with herniated lumbar disc is related to position of herniation rather than magnitude and exent of herniation. Inflammed intraspinal tissues are significantly associated with low back pain and sciatica.

The relationship between hypermethylation of CpG islands in the promoter regions of O6-methylguanine DNA methyltransferase (MGMT) genes and laryngeal squamous cell carcinoma was explored. Methylation-specific PCR and semi-quantitative RT-PCR were used to study the promoter methylation and mRNA expression of the MGMT gene in laryngeal carcinoma tissues, tissues adjacent to the tumor and normal laryngeal tissues. Hypermethylation of MGMT gene was detected in 16 samples of 46 (34.8%) laryngeal squamous cell carcinoma samples. However, the MGMT hypermethylation was not detected in all tissues adjacent to the tumors and normal tissues. No significant difference in MGMT gene hypermethylation was found in samples with different histological grades (chi2 = 3.130, P = 0.077) or in samples from patients with different TNM status (chi2 = 3.957, P = 0.138). No expression of MGMT mRNA was detected in all hypermethylated laryngeal carcinoma tissues. The expression of MGMT mRNA was detected in all unmethylated laryngeal carcinoma tissues, tissues adjacent to the tumors and normal tissues. It suggests that MGMT gene promoter hypermethylation is associated with MGMT gene transcription loss in laryngeal carcinoma tissues and possibly plays an important role in carcinogenesis of laryngeal tissues.
Carcinoma, Squamous Cell/*genetics ; CpG Islands/genetics ; DNA Methylation ; DNA Repair ; Laryngeal Neoplasms/*genetics ; O(6)-Methylguanine-DNA Methyltransferase/*genetics ; Polymerase Chain Reaction/methods ; Promoter Regions (Genetics)/*genetics