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.

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 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 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 .

BACKGROUND:There is often space between implant and bone during immediate implantation.Whether biological membrane is needed to guide bone regeneration remains poorly understood.OBJECTIVE:To createdifferent sizes of space between femurand implantsindogs and to observe the effects of biological membrane on bone regeneration capacity of bone defects surrounding implants.DESIGN,TIME AND SETTING:A self-control animal experiment was performed at the Laboratory Animal Center,Norman Bethune College of Medicine,Jilin University and School of Stomatology,Jilin University between March and December 2005.MATERIALS:BLB hydroxyapatite-coated implant was provided by Beijing Leiden Biomaterial Co.,Ltd.,China;BME-10X collagen membrane was purchased from Fujian Better Biotechnology Co..Ltd.,China.METHODS:BLB implants were installed in the bilateral proximal femoral bone to create standard gradient bone defects with horizontal width 3 mm.vertical depth 5 mm,and horizontal lengths of 0,1,2,3,and 4 mm Bone defects on the left femur were sutured directly and those on the right femur were covered with biological membrane prior to suture.All animals were sacrificed at 3 months after surgery.Specimens containing implants were harvested to prepare tissue blocks for radiological observation.MAIN OUTCOME MEASURES:The quantity,color,and texture of newly formed bone surrounding implants were observed from the surface and profile levels.The implant-bone integration and new bone formation were also examined by soft X-ray photography.RESULTS:Grossobservation results revealed that when the horizontal length of bone defect was 3 mm or less,there was no significant differenee in bone density between the newly formed bone and the host bone no matter whether biological membrane existed or not;when the horizontal length of bone defect was 4 mm the bone density was better when biological membranes were used than not.Soft X-ray photography results revealed that when the horizontal length ofbone defect was 3 mm or less.no significant difference in bone density and bone trabecular morphology and orientating was found between newly formed bone and host bone no matter whether biological membrane was used or not;in the 4-mm-length bone defect areas.implants contacted with newly formed bone directly,but the calcified degree ofnewly formed bone was poor,bone trabecula was thin,and bone trabecular course was irregular,nevertheless,the calcified degree of newly formed bone was better under the condition of being with biological membrane than without biological membrane.CONCLUSION:Biological membrane exhibits strong capacity to promote the regeneration and repair of bone defect tissue with a horizontal length of 3 mm or less,and plays an important role in repatr of large sizes of bone detect