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OBJECTIVE@#To construct an adenoviral vector that codes for both human NT3 and EGFP, to confirm the transduction efficiency in rat cochlear cultures and to assess the protection of NT3 on SGNs survival.@*METHOD@#PAdeasy-1 and pAdTrack CMV were used to constructed Ad/NT3 adenovirus and then to transfer postnatal day 3 rat cochlear cultures. The transduction efficiency was determined by microscope observation. The amounts of SGNs were counted to evaluated protection of Ad/NT3 on SGNs survival.@*RESULT@#EGFP positive cells were observed in all cochlear turns. There was approximately 49% in outer sulcus cells and 27% in the interdental cells; less than 2% of the hair cells and SGN. The amounts of SGN of treated Ad/NT3 adenovirus are more than cochlea SGN only Ad/EGFP adenovirus after cultured for 15 days.@*CONCLUSION@#Ad/NT3 adenovirus could transduce EGFP and NT3 in large number of supporting cells, but few hair cells or SGNs. The putative release of NT3 from these supporting cells could enhance cell survival and promote neurite outgrowth from SGNs.
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Animals , Humans , Rats , Adenoviridae , Genetics , Basilar Membrane , Cell Biology , Cell Survival , Genetics , Cells, Cultured , Cochlea , Cell Biology , Genetic Vectors , Hair Cells, Auditory , Cell Biology , Neurotrophin 3 , Genetics , Rats, Inbred F344 , TransfectionABSTRACT
OBJECTIVE@#To establish a practical model for Wistar rat cochlea organ culturing in vitro, and to observe the growing status in hypoxia of the spiral ganglion cell and nerve fiber.@*METHOD@#We used an in vitro hypoxia model and dissociated cultures of the basal membrane from the cochlea of 3-day-old Wistar rats. And put them in incubator (37 degrees C, 90% N2, 5% CO2, 5% O2) to hypoxia culture for different times. The culture were Immunofluorescence dyed and count the number of the spiral ganglion cell and the cell density in unit area (24 mm x 36 mm), and observe the morph of nerve fiber under the confocal microscope, the results were compared with controls.@*RESULT@#Hypoxia early (6 h) nerve fiber appear edema, spiral ganglion cell didn't change compared with controls; nerve fiber appear break and disintegration and the spiral ganglion cell decrease in 12 hours culturing, and the cell density in unit area had remarkable difference compared with control (P < 0.01). Hypoxia leads to the cell density decrease in a time-dependent manner, the longer of cultures times in hypoxia, the heavier of damage in spiral ganglion cell and nerve fiber. Twelve hours culturing, and the cell density in unit area had remarkable difference compared with control (P < 0.01). Hypoxia leads to the cell density decrease in a time-dependent manner, the longer of cultures times in hypoxia, the heavier of damage in spiral ganglion cell and nerve fiber.@*CONCLUSION@#The study findings suggest that hypoxia makes the spiral ganglion cell and nerve fiber damage of culturing in vitro, and nerve fiber more susceptible than spiral ganglion cell for hypoxia.
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Animals , Female , Male , Rats , Cell Count , Cell Survival , Hair Cells, Auditory, Inner , Cell Biology , Pathology , Hypoxia , Pathology , In Vitro Techniques , Nerve Fibers , Pathology , Rats, Wistar , Spiral Ganglion , Cell Biology , PathologyABSTRACT
Objective To observe the effect of Ixeris sonchifoila,a kind of Chinese traditional medicine on noise-induced hearing loss.Methods Fourty-two guinea pigs were divided into two groups.The animals were exposed under 110 dB(n=21) and 120 dB(n=21) noise,separately.Each group was divided into medicine group(n=7),saline group(n=7) and control group(n=7).Ixeris sonchifoila was used in the medicine group and saline in the saline group immediately after the noise exposure by abdomen injection.The threshold was recorded 7 and 14 d later by ABR.Cochlea patch was performed to find hair cell loss.Results By using Ixeris sonchifoila,the threshold shift was significantly reduced(P
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OBJECTIVE To investigate the role of TLR4 signaling pathway in secretory otitis media of rats.METHODS Rats SOM models were made by infusing FS and LPS to the middle otocysts. The rats were killed separately at the 3rd, 5th, 7th, 9th,11th and13th day(5 rats each time). And then the expressions of TLR4, iNOS, IL-8mRNA of middle ear mucosae were detected. NF-?B was also detected through the method of immunohistochemistry. RESULTS The expressions of TLR4mRNA, iNOS and IL-8mRNA in middle ear mucosae of SOM group were significantly higher than that of control group. Immunohistochemistry method showed that the activation of NF-?B in middle ear mucosae was obviously increased in SOM. CONCLUSION The expression of TLR4 was obviously increased in SOM and it activates the NF-?B, promotes the expression of downstream IL-8. That maybe the mechanism which cause SOM.
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OBJECTIVE To evaluate the effectiveness of ~(125)I seeds brachytherapy in advanced head and neck malignant tumor.METHODS Twelve patients with post-radiotherapy recurred laryngeal cancer with bilateral cervical lymph node metastasis, tongue cancer with bilateral cervical lymph node metastasis or recurred maxillary sinus cancer and so on were operated radically,and ~(125)I seeds were implanted in wound area during operation.RESULTS With a follow-up of 24 to 32 months,no tumor recurred in 11 patients,and only one patient with tongue cancer recurred at 6 month after operation.No local infection, acute radioactive lesion,vessel broken or seeds migration happened.CONCLUSION The ~(125)I brachytherapy can be used as one of the comprehensive treatment method in advanced head and neck malignant tumors,especially in the recurred tumors after operation or radiotherapy or in the cancer which can not be radically removed.It's also an effective method to improve local control in the subset of patients with doubtful positive margins.
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Objective To investigate the correlation of cochlear injury and expressions of Bcl-2 and Bax in the cochlea of diabetic rats.Methods Thirty five Wistar rats were randomly divided into diabetic group(n=25) and control group(n=10).Diabetic deafness was induced in rats by intraperitoneal injection of streptozotocin,while the rats in normal group were injected with normal saline.Immunohistochemical staining was used to detect the protein levels of Bcl-2 and Bax in the cochlea after 3 months.Results No significantly morphological change in cochlea of diabetic rats was found.The Bcl-2 levels in stria vascularis in diabetic group and control group were 135.32?21.69 and 45.32?9.08,respectively,there was significant difference(P
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0.05).IHC loss was found under hypoxia for 2 h to 48 h in a time dependent manner,the cell density in unit area had remarkable difference compared with control(P
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BACKGROUND: The extracellular signal-related kinase(ERK) cascade is activated by its specific upstream kinase MAPK/ERK kinase(MEK) and identified as a critical regulator of cell growth, memory formation and long-term neuronal plasticity.OBJECTIVE: To investigate the role of ERK activation in establishment and maintenance of chronic neuropathic pain.DESIGN: A randomized controlled experiment based on the animals.SETTING: Department of Anesthesiology of a university-affiliated hospital.MATERIALS: The experiment was performed in the Medical Institute of Anesthesiology of Jiangsu Province from September 2003 to June 2004. Totally 115 male clean SD rats weighting 220 to 300 g were provided by the Experimental Animal Center of Xuzhou Medical Colleg e. The rats were raised in separate cages with natural lighting at 20 to 25 ℃, having free access to food and water.INTERVENTIONS: Five days after chronic constrictive injury(CCI) model was established, different doses of U0126 were intrathecally injected according to Mestre' s method. The control group received sterile intrathecal injection of 50 g/L dimethylsulfoxide.hyperalgesia by using von Frey filaments and thermal hyperalgesia stimulator activation and translocation of ERK in spinal cord dorsal horn by immunohistochemistry and Western blot analysis.RESULTS: CCI could increase the activity of EPK in spinal cord dorsal horn. Intrathecal injection of U0126 significantly attenuated CCI-induced mechanical and thermal hyperalgesia.CONCLUSION: Activation and translocation of ERK contribute to formation and maintenance of CCI-induced neuropathic pain.
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Objective:Mutations in the CX(connexin)26 gene (GJB2), encoding the gap\|junction protein. Connexin 26, have been shown to be the major cause of non\|syndromic recessive deafness. Among these mutations, the CX 26 cDNA (35delG) mutation accounts for the majority of this kind of deafness are European and America, but it does not present in China. Therefore, we intend to investigate the mutation hot spot of connexin 26 in China deafness populations. Mathods:The mutation of 233delC was detected by the technique of PCR\|RFLP and analyzed by directly sequencing protocols in 219 patients with kinds of deafness and 50 individuals with normal hearing. Results: The frequency of the 233delC mutation was 47/219(21.5%) in the Chinese population. No mutation was found in group with normal hearing and dominant nonsyndromic hearing loss. The mutation frequency for 233delC was 4 in 15 patients (26.7%) with genetic prelingual deafness and 10 in 50 patients(20.0%) with drug\|deafness. Conclusion: The 233delC mutaion had a high frequency in Chinese population with hearing loss, especially congenital deafness. Our data indicated that specific combinations of GJB2 mutation exist in different populations.
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Objective:To study the methods of early qualitative diagnosis of laryngeal displasia and earlycarcinoma. Methods:We investigated displasia (34 cases),squamous cell carcimoa (19 cases) and vocalcord polyps (17 cases) of larynx which were embeded in paraffin by PCR－SSCP analysis of p53 gene,im－munochemi stain of p53 protein and Ⅳ collagen,cytometer (determining fraction of cell cycle) and AgNORstain. Results :Eleven of 34 cases of displasia were all positive in the indexes. Seven of 11 cases were alsopositive in pathology,but others were negative. Five of 34 cases of displasia were all negative in the index-es,2 of 5 cases were positive and 1 of 5 cases were negative,other two cases did not have following-up re-sults in pathology. Conclusion :The results suggest that it is the combination of the morphological indexeswith biological indexe that is more scientific and more accurate in quatitative diagnosis of premalignantchange.