To explore the mechanism of therapeutic effect of basic fibroblast growth factor (bFGF) in the treatment of blast-induced deafness, and to define its optimal clinical use, bFGF was infused into the guinea pig's cochlea, combined with intramuscular injection of bFGF after being exposed to explosion. The compound action potential (CAP) and auditory brainstem response (ABR) were measured in these animals. 125I labeled basic fibroblast growth factor ( 125I -bFGF) was injected intraperitoneal to the guinea pigs to observe whether it could pass through the blood-labyrinthine barrier. The results showed that bFGF infused to the cochlea might facilitate recovery of hearing loss following acoustic trauma. Basic fibroblast growth factor ( 125I -bFGF) intraperitoneally injected, could not pass through the blood-labyrinthine barrier. However intramuscular bFGF promoted the recovery of hearing, probably indirectly through the neuro-immunity network.

To explore the spatiotemporal patterns of the neuronal excitatory propagation in vestibular nucleus, brainstem sections were prepared from postnatal 1～5 day mice, and stained with RH155, which was an light absorbent voltage sensitive dye, for 20 minutes. A multiple site optical recording system was used for optical imaging of the evoked responses after electrical stimulation of the vestibular nerve. After stimulation of the vestibular nerve, optical responses were revealed in the vestibular nucleus. There was propagation of excitation in both ipsilateral vestibular nucleus and contralateral vestibular nucleus after ipsilateral vestibular nerve stimulation. These optical signals were wave length dependent. The optical signals consisted of two components: the spike like fast signal and long duration slow signal. All the responses were abolished by 20?mol/L tetrodotoxin (TTX). The effect of TTX was irreversible. The slow signals were entirely eliminated after the application of Ca 2+ free solution. The effect of Ca 2+ free solution was reversible. These results suggested that the slow signal might be postsynaptic excitation potential. The present study indicated that the use of optical recording to reveal visually the synaptic transmission of afferent input in vestibular nucleus in the brainstem was feasible.

Objective To explore the spatiotemporal patterns of the glutamatergic transmission in vestibular nucleus. Methods The brainstem slices were prepared from postnatal 1-5-day mice. The slices were stained with RH155, which was a light-absorbent voltage-sensitive dye, for 20 min. A multiple-site optical recording system was used for optical imaging of the evoked responses after electrical stimulation to the vestibular nerve. Results The spatiotemporal patterns of excitatory propagation in VN were illustrated. The effects of glutamate antagonist in VN after being bathed in APV (100mol/L, NMDA receptor antagonist) or CNQX (10mol/L, non-NMDA receptor antagonist) were observed in the postnatal 1 to 3 day-mouse brainstem slices. Our data showed that the percentage of APV sensitivity ranged from 50% to 84.2%, with a mean of 64.9%?9.06% (n=18). The percentage of CNQX sensitivity ranged from 15.8% to 50%, with a mean of 35.1%?9.06% (n=18). Conclusion The study indicated that the use of optical recording for revealing visually the synaptic transmission of afferent input in VN in brainstem was feasible. Both NMDA and non-NMDA receptor were sensitive to the neuronal transmission of EPSP in VN, but the NMDA sensibility to EPSP was higher than that of non-NMDA in newborn mice.

Objective To investigate the structure and function of cross-links on stereocilia of guinea pig outer hair cell (OHC). Method The ultrastructures of cross-links on stereocilia of guinea pig OHC were observed by scanning electron microscopy and tannic acid procedures. Results The side-links ran horizontally between OHC stereocilia. The stereocilia of the same and different row on each hair cell were joined by horizontally-running links. The side-links were observed in the first turn as well as the fourth turn. There were more side-links on the intact stereocilia than that on the disrupted arrangement. The side-links in the first turn and the fourth turn were more abundant than that in the second turn and the third turn. The side-links between stereocilia were spare if the stereocilia were separated. The number of side-links on stereocilia was proportional to the number of bulb-like structures on stereocilia. Conclusion The side-link is a kind of morphological structure on hair cells stereocilia of cochlea. The results suggest that cross-links play an important role in maintaining the structure and function of the hair cell stereocilia.

ObjectiveTo investigate the prevalence and distribution of 16S rRNA methylase gene and research the relationship with drug resistant spectrum.And preliminary explore its role in molecular epidemiology analysis.MethodsCollected 69 clinical isolates of non repetitive ESBL-producing Klebsiella pneumoniae in our hospital from Mar to Sep 2010.Detection 16S rRNA methylation enzyme gene by PCR,and analyze ESBL genetype and integron gene of the positive strains.All PCR products were sequenced for determination.Plasmid conjugation test and plasmid elimination method to determine dissemination of 16S rRNA methylase gene.Then we used ERIC-PCR genotyping technology for the establishment of DNA fingerprinting.ResultsIn sixty-nine strains,twenty isolates were rmtB positive (28.9％),two isolates were armA positive,and two strains coproduce rmtB and armA.All positive isolates carried the CTX-M gene,detemined by sequencing,14 strains of CTX-M-14 gene,6 strains of CTX-M-15 gene,14 strains carried TEM1 gene,8 strains carried SHY gene,sequencing showed that 5 strains of SHV-12 gene,3 strains of SHV-11 gene,3 strains carried OXA-10 gene,3 strains carried VBE-1 gene.In addition,the intl gene was found in 12 isolates of 20 rmtB positive strains.All the intl gene positive strains were divided into five kinds gene cassettes,which contained drfA25,drfA1,drfA12,aadA1,aadA2,sat and blaVEB-1 genes.Respectivily,16S rRNA methylase gene positive strains were divided into five genetypes using ERIC-PCR technology.A genetype was the advantage popular clones.Conjugative plasmid and elimination test found that rmtB gene was located in a plasmid in KP5 and KP16 isolates with A genetype,and can disseminate by conjugation.ConclusionA high prevalence of 16S rRNA methylase gene-rmtB was found among clinical ESBL-producing K.pneumoniae isolates in our hospital,which could lead to resistant to almost all aminoglycoside at a high level.Both horizontal gene transfer and clonal spread were responsible for the dissemination of the rmtB gene.In addition,K.pneumoniae co-producing ESBLs,16S rRNA methylation enzymes and class Ⅰ integron existed and were spreading.

Objective To investigate molecular epidemiology and antimicrobial susceptibility of carbapenem-resistant strains of Klebsiella pneumoniae isolated from children.Methods From July 2010 to June 2011,twelve non-replicate clinical isolates of carbapenem-resistant Klebsiella pneumoniae were consecutively collected from children inpatients in the Second Hospital of Wenzhou Medical Colloge.All of the isolates were identified by the automated microbiology systems.Modified Hodge test was used to screen strains producing carbapenemases.Pulsed field gel electrophoresis(PFGE) was performed to analyze the homogeneity of genomic DNA of Klebsiella pneumoniae.KPC,IMP,GIM,SPM,SME,OXA-10,bla(s),VIM gene and integrase gene were amplified by PCR and then sequenced to cofirm the genotypes;Plasmid conjugation experiment was used to study the transfer method of bacterial resistance.Plasmid-curing test were used to initally locate the resistant genes.Results One(8.3％),5(41.7％),7(58.3％),1(8.3％),1(8.3％) and4(33.3％) of12isolates were susceptible to gentamicin,tobramycin,amikacin,ciprofloxacin,levofloxacin and trimoxazole,respectively.All isolates carried KPC-2,TEM-1 and SHV genes(six for SHV-11-like,six for SHV-12-like).Eleven of twelve isolates with KPC-2 gene carried CTX-M genes(4 for CTX-M-14-like,6 for CTX-M-15-like).Two isolates carried OXA-10 genes,and one isolates carried PER-1 gene.None of NDM-1,GIM,SPM,SIM and VIM carbapenemase genes was detected in 12 isolates.All of 12 isolates carried Int 1 genes.The plasmids of 2 isolates were transgerred into the recipients E.coli EC600.PCR and sequence analysis revealed that blaTEM-1 and blaCTX-M-15-like were co-transferred with the KPC-2 gene to the recipients.Elimination of KPC-2-encoding plasmid from Kp7 and Kp12 resulted in imipenem susceptibility in the two isolates.Amplification revealed that KPC-2 gene was lost by the plasmid-curing test.Of the 12 isolates,5 patterns were obtained by PFGE.Pattern B and C were the main drug resistant clones.Conclusion KPC-2 gene are the major carbapenemase genes in Klebsiella pneumoniae isolated from children,including ESBLs and integrase.Some resistance genes can be disseminated by plasmids.

Diagnosis related groups (DRGs)have been widely used in many countries and regions.The versions of DRGs in different regions are modified according to the local circumstances and conditions.Beijing version DRGs (BJ-DRGs)is the first DRGs system in Chinese,which was developed locally based on the conditions of medical information system and health relative policies in Beijing.This paper introduced the grouping principles,logic and methodology of BJ-DRGs.Taking“ear,nose,mouth and throat disorders”for example,this paper demonstrated the grouping process of BJ-DRGs.

Objective To show the capability of NF-κB expression in cochlea. Methods Kanamycin (KA) was subcutaneously injected twice daily for 3 and 7 days with an eight hours interval between two injections, and lipopolysaccharide (LPS) was injected into the tympanic cavity of mice. Equal amount of saline was injected for 7 days as control. Frozen sections of all mice cochleae were examined immmunohistochemichally with rabbit polyclonal NF-κB p50. Expression of NF-κB p50 immunoreactivity of mouse cochleae is identified as showed as brown reaction products characteristic of DAB immunohistochemistry. Results Immnoreactivity NF-κB p50 in mouse cochlea was localized in the organ of Corti, spiral limbus, tectorial membrane, the vascular stria, spiral ligament, spiral ganglion and nerve fibers. The immunoreaction could be observed in all spirals throughout the cochlea. Stronger staining was visible in spiral ligament, tectorial membrane, spiral prominence, spiral ganglion and nerve fibers, and the organ of Corti. The immunoreaction in the vascular stria was weaker than that in the structures mentioned above. The immunoreaction in the organ of Corti was observed in inner hair cells (IHC) and outer hair cells (OHC), inner and outer pillar cells, Deiter's cells, and Boettcher's cells. The immunoreaction was weaker in inner sulcus cells, Hensen's cells and Claudius'cells. The stronger immunoreaction was observed in nucleus of spiral ganglion cells. The nucleus of IHC and OHC remained unstained. Conclusion The injection of LPS/KA can promote NF-κB p50 expression to induce an acute reaction in mouse cochlea.

Objective:To probe the method of isolating outer hair cells (OHC) from each of four turns of the guinea-pig cochlea. Method:From eight guinea pigs the organ of Corti from each of four turns of the cochlea were dissected, and then treated using enzyme. Result:A fair amount of living OHCs from each of four turns were obtained. The length of OHCs from each of four turns were 23.81,34.50,60.48 and 71.37 μm. Conclusion:The key to success in isolating OHCs from each of four turns of the cochlea is to know very well the anatomical characteristics of each of four turns of the cochlea and be operated in accordance with normal rules.

Objective:To investigate the relationship between herpesviridae and malignant or benign laryngeal diseases.Method:128 paraffin-embedded laryngeal squamous cell carcinoma and laryngeal epithelium hyperplastic lesions were detected by polymerase chain reaction (PCR) and PCR-ISH for herpesviridae. Result:HSV-1 was detected in 10 cases by PCR,among them 3 were laryngeal squamous cell carcinoma (LSCC),1 was carcinoma in situ(CIS),4 were laryngeal polyps and 2 were laryngeal keratosis. Except 1 LSCC and 1 CIS, 8 of 10 cases were positive while detected by PCR-ISH. In benign diseases, signals were shown from basal layer to superficial cell; in malignant lesions, the signals were scattered in the diseases.Conclusion:Most of laryngeal diseases were not related to herpesviridae, but HSV-1 may acts as initiator in the development of a few cases.