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.

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.

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 the relationship between the mechanism of gynecomastia and serum hormone levels ,as well as liver function in male patients with liver cirrhosis .Methods Forty‐six male patients with liver cirrhosis and gynecomastia were collected as gynecomastia group from March 2013 to March 2014 ,and at the same period seventy male patients with liver cirrhosis but without gynecomastia were studied as non‐gynecomastia group . The condition of mammogenesis and maximum of breast thickness were measured by bilateral breast ultrasound .Hormones including luteinizing hormone (LH) ,follicle‐stimulating hormone (FSH) , prolactin (PRL) ,estradiol (E2) ,progesterone (PRGE) ,and testosterone (T);liver function including alanine aminotransferase (ALT) ,aspartate aminotransferase (AST) ,total bilirubin (TBil) and serum albumin (Alb);blood coagulation function including prothrombin time (PT) ,platelet count (PLT) were examined and the Child‐Pugh scores were calculated .t‐test was performed for results comparison between gynecomastia group and non‐gynecomastia groups .Chi‐square test was used to compare the difference in drinking rate between two groups . The patients of gynecomastia group and non‐gynecomastia group were further divided into Child‐Pugh Grade A ,B and C subgroups according to Child‐Pugh scores and the patients of gynecomastia group were divided into subgroups according etiology such as posthepatitic cirrhosis ,alcoholic liver cirrhosis and posthepatitic cirrhosis combined with alcoholc cirrhosis .Single factor analysis of variance was applied to compare the laboratory findings between subgroups ,and least‐significant difference mothod was used to further compared the differences between two subgroups .Results Among forty‐six male patients with liver cirrhosis and gynecomastia ,the mean thickness of breast was (7 .56 ± 2 .84) mm .All the differences of TBil ,Alb ,PT and Child‐Pugh score of Child‐Pugh grade patient were statistically significant between gynecomastia group and no gynecomastia group ((96 .72 ± 75 .86)μmol /L vs (60 .57 ± 54 .00)μmol /L ,(29 .12 ± 4 .90) g/L vs (33 .86 ± 6 .86) g/L ,(19 .06 ± 4 .76) s vs (15 .54 ± 2 .57) s ,11 .54 ± 0 .91 vs 10 .33 ± 0 .57 ,respectively ,t=2 .79 ,-4 .33 ,4 .58 ,2 .22 ,all P<0 .05) . The alcoholic drinking rate of gynecomastia group (74% (34/46) vs 53% (37/70)) increased ,and the difference was statistically significant compared with that of non‐gynecomastia group (χ2 =5 .183 , P<0 .05) .There was no statistically significant difference in E2 levels between gynecomastia group and non‐gynecomastia group (P>0 .05) .PRL and E2/T ratio ((404 .49 ± 297 .26) mU/L and 68 .74 ± 46 .37) were higher than those of non‐gynecomastia group ((279 .77 ± 111 .57) mU/L and 13 .60 ± 11 .55) ,and T was lower than that of non‐gynecomastia group ((7 .15 ± 5 .74) nmol/L vs (15 .46 ± 8 .53) nmol/L) ,and the differences were statistically significant (t=2 .72 ,7 .90、-6 .27;all P<0 .05) .Among patients with gynecomastia ,breast of patients with alcoholic liver cirrhosis was significantly thicker than that of patients with posthepatitic liver cirrhosis ((9 .25 ± 3 .59) mm vs (6 .67 ± 2 .48) mm) ,while the level of PRGE was lower than that of patients with posthepatitic liver cirrhosis ((0 .61 ± 0 .51 ) nmol/L vs (1 .49 ± 1 .47 ) nmol/L ) , and the differences were statistically significant (F= 3 .634 and 2 .674 ,both P< 0 .05) .Along with the severity of liver injury ,E2 level of gynecomastia group gradually increased ,however there was non‐statistically significant difference compared with non‐gynecomastia group (P>0 .05) .T level of gynecomastia group gradually decreased ,and those of Child‐Pugh B ,C subgroup ((8 .20 ± 7 .58) nmol/L and (4 .18 ± 3 .76) nmol/L) were siginificantly lower than that of Child‐Pugh A subgroup of non‐gynecomastia group ((17 .64 ± 9 .04) nmol/L ,F=9 .37 ,P<0 .05) .The E2/T levels of gynecomastic group gradually increased .There was significant difference in E2/T level between Child‐Pugh C subgroup of gynecomastia group (105 .49 ± 94 .42) and Child‐Pugh A grade subgroup of non‐gynecomastia group (11 .38 ± 9 .60 ,F=12 .57 ,P<0 .05) .Conclusions There are different degrees of sex hormone disorder in the serum of male patients with liver cirrhosis and gynecomastia which is more significant in PRL ,T and E2/T .T and E2/T level are correlated with the degree of liver functional impairment .Gynecomastia in alcoholic liver cirrhosis is more severe than that of posthepatitic liver cirrhosis .

This paper presented the hereditary pattern of otosclerosis in families. Among 1500 cases there were 13 patients, belonging to 6 families, and 20 ears were surgically treated. There was often a marked similarity in clinical features of affected siblings within the same family.The clinical characteristics are as following: (1) The age at inset of hearing loss seems similar. There is a difference of 1 or 2 years in two families znc 3 to 10 years in the other four. (2) The side of first hearing loss is the same in affected individuals of 6 families. (3) The propositi are females in 5 families (83.3%). (4) Auditory curve of hearing loss is similar in 4 familiers, and different in the other two. (5) The footplate lesions are similar in macroscopic features, thin footplate in two fimalies and thick footplate in the other four.

Objective To investigate the anti-tumor effects of total saponins,extracted from stems and leaves ofParispolyphyllavar.yunnanensis. Methods Taking mouse stomach carcinoma MFC cell line,human mammary cancer cell line(MCF-7)and human cervical carcinoma cell line(Hela),and their tumor-bearing mice as models,the antitumor activity was carried out in vitro and in vivo. CCK-8 assay was used to determine the inhibitory effect in vitro. The tumor-bearing mice model was induced by tumor cell vaccination in normal mouse forelimb left axillary subcutaneous and these mice were randomized into NS group,cytoxan group(30 mg/kg),saponins high-dose,medium-dose and low-dose groups(60,30 and 15 mg/kg). They were given intraperitoneal injection once a day for consecutive 15 days. The tumor inhibition rate and survival of the tumor-bearing mice were measured. Results Saponins,extracted from both aboveground and underground ofP.polyphyllavar.yunnanensis could significantly inhibit the growth of MFC,MCF-7 and Hela cells in time- and dose-dependent manners. The tumor weight in each drug treated group was significantly lower than that in the control group. Conclusion Saponins,extracted from both aboveground and underground ofP.polyphyllavar.yunnanensis have antitumor activity.

Objective The purpose of this study was to edit and psychometrically equate a set of Mandarin bisyllabic word lists. Methods 964 bisyllabjc words were recorded by male talker of standard Mandarin,352 words were selected to compose 10 lists. Percentage of correct word recognition was measured for each word at four intensity levels using 20 normal hearing subjects. The order of the presentation of the lists was randomized for each subject. U-sing Statistica7.0 Performance-Intensity function for each word was plotted,slopes and thresholds of them were calculated. 242 words were chosen after that, The thresholds and slopes of these words were in Guassian distribution. These words were ligitally adjusted and included in six Mandarin bisyllabic word lists of 40 words each. Two of them were for practice, four of them for test. 36 subjects with normal hearing served in our equivalent test. The orders of the presentation of the lists were randomized far each subject and results were expressed as thresholds. Results Two-factor ANO-VA was used to compare the thresholds of the 4 lists, F=1.978,P = 0.209. Conclusion These lists were considered to be primarily equivalent with each other.