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Toll-like receptor 5 (TLR5) is responsible for the recognition of bacterial flagellin in vertebrates. In the present study, the first TLR5 gene in duck was cloned. The open reading frame (ORF) of duck TLR5 (dTLR5) cDNA is 2580 bp and encodes a polypeptide of 859 amino acids. We also cloned partial sequences of myeloid differentiation factor 88, 2'-5'-oligoadenylate synthetase (OAS), and myxovirus resistance (Mx) genes from duck. dTLR5 mRNA was highly expressed in the bursa of Fabricius, spleen, trachea, lung, jejunum, rectum, and skin; moderately expressed in the muscular and glandular tissues, duodenum, ileum, caecum, and pancreas; and minimally expressed in the heart, liver, kidney, and muscle. DF-1 or HeLa cells transfected with DNA constructs encoding dTLR5 can activate NF-kappaB leading to the activation of interleukin-6 (IL-6) promoter. When we challenged ducks with a Herts33 Newcastle disease virus (NDV), mRNA transcription of the antiviral molecules Mx, Double stranded RNA activated protein kinase (PKR), and OAS was up-regulated in the liver, lung, and spleen 1 and 2 days post-inoculation.
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Animals , Humans , 2',5'-Oligoadenylate Synthetase/genetics , Cell Line , Cloning, Molecular , Ducks , Gene Expression Regulation/physiology , Immunity, Innate , Myeloid Differentiation Factor 88/genetics , Myxovirus Resistance Proteins/genetics , Newcastle Disease/metabolism , Newcastle disease virus/classification , RNA, Messenger/genetics , Species Specificity , Toll-Like Receptor 5/geneticsABSTRACT
OBJECTIVES: Apoptosis may play an important role in the mechanism underlying the GJB2 gene conditional knockout (cCx26) mice cochlear cell death. The objective of this study was to explore the the damage mode of the outer hair cells (OHCs) and its real time point of apoptosis and provide information to further explore the role of apoptosis in the happening of hearing loss in cCx26 mice. METHODS: Cochleae from mice at various developmental stages (P8, P12, and P21) were dissected out and first used to be observed under the scanning electron microscope (SEM). Basilar membranes from mice at P8, P14, P18, and P21 were stained by fluorescein isothiocyanate-conjugated phalloidin and propidium iodide (PI) and examined under confocal microscope. RESULTS: The loss of OHCs of cCx26 knockout mice was first set between P12 and P21 under SEM. Whole mount phalloidin and PI staining revealed that obvious apoptotic appearance of the OHCs surface morphology was observed at P18. CONCLUSION: Typical apoptotic morphology was found in the OHCs in the organ of Corti of the cCx26 mice at P18. This may provide information to further study the role of apoptosis in the occurrence of hearing loss of cCx26 mice.
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Animals , Mice , Apoptosis , Basilar Membrane , Cell Death , Cochlea , Connexins , Electrons , Fluorescein , Hair , Hair Cells, Auditory, Outer , Hearing Loss , Hearing Loss, Sensorineural , Mice, Knockout , Organ of Corti , Phalloidine , PropidiumABSTRACT
Objective To study and summarize the tumor-free technique points for coordinating the operations aiming at rejecting aggressive spinal tumors by microwave and hyperthermia induced method. Methods 12 cases of patients with malignant bone tumors were performed with operations for rejecting tumor, using tumor-free technique to avoid cross infection, protect normal tissue nearby and prevent complications effectively. Results 12 cases of operations were successfully performed, no metastasis Was seen, no adjacent tissue injury and peri-operative complications occurod. Conclusions Microwave-induced hyperthermia in management of aggressive spinal bone tumors can increase the post-operative living-quality of patients, make both medical staff and patients satisfied and reach anticipated effect.
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Objective To evaluate the safety of Ad-Mathl administration to inner ear and provide the base data for vestibular dysfunction gene therapy.Methods Ten mature Wistar rats were divided into normal control group(srats) and adenovirus(E1,E3-Deleted and carried mathl and enhanced green fluorescent protein report gene,Ad-Mathl-EGFP)scala vestibuli transfer group(5 rats).Right ears of the Ad-Mathl-EGFP transfering group rats were deliveried 5ul Ad-Mathl-EGFP(physieal tite 2.1 10~(11)v.P./ml)into cochleas through the way of drilling scala vestibuli of cochlear basal turn.As a control,the normal group received nothing to inner ear.In order to estimate functional condition of vestibule and cochlea,the click-evoked potentials on the surface of the cervical dura mater(CDM-CEP),auditory brain stem response(ABR)and swimming time were recorded in all rats at 7 days after treatment,and then histologic and morphologic observation were carried out after animals were sacrificed.Results All animals' morphologic observation showed that inner ear hair cells were normal after transfer.Seven days after transfer,the swimming time was 4.0±0.71 s in normal control group and 5.0±0.71 s in scala vestibuli delivery group.The threshold of CDM-CEP and ABR were 85±3.54 dB SPL and 37±4.47 dB SPL in normal control group,and 89±6.52 dB SPL and 40±3.54 dB SPL in Ad-Mathl-EGFP scala vestibuli delivery group,respectively.There was no significant difference existed between control group and Ad-Mathl-EGFP scala vestibuli delivery group.Conclusion The Ad(E1,E3-Deleted)is safe for vestibular and cochlea hair cells and can be used as an ideal vector of gene transfer.
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OBJECTIVE To standard operating room hospital infection monitoring management,to guarantee the management effect of perioperation hospital infection.METHODS Based on standardized,systematized,sequenced theory and idea.The operating room hospital infection supervisory system plan were for mulated.Including:hospital infection monitoring of operation room,test result analysis,and monthly report system.RESULTS We had grasped the key the link,during operating room scale expansion.The prompt establishment perfect special control system,monitoring the perioperation hospital infection effectively,received highly praise.CONCLUSIONS Sysmatic mornitoring and scientific control and good adaptive team.Can guarantec continualls improvement.
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Objective To assess the feasibility of adenoviral vectors mediate cochlear gene transfer by postau-ricular microinjection through the round window membrane in mouse. Methods Twelve 5-week old C57BL/6J mice were selected for the study: 8 were implanted with Ad-EGFP by postauricular microinjection through the round window membrane, and 4 with artificial perilymphatic fluid. On postoperative days 5 and 14, the animals were sac-rificed and the surface preparation of cochleae was observed. Results Two animals died after operation. Bright green fluorescence in the cochleae was observed in Ad- EGFP groups. Gene expression on day 14 after operation was higher than that on day 5. However, the control group was free of fluorescence. Oonclusion The postauricular route of the cochlear gene transfer in mice is simple to operate with little side-effect. The technique of transgenic delivery into the inner ear through RWM by mieroinjection is feasible and effective.
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OBJECTIVE To find out the status of and impact factors on bacterial concentration in clean operating room.METHODS Set a questionnaire to the staff in the operating room of 24 tertiary 3A hospitals.RESULTS The 9 basic questions got correct and accordant answers.The answers for the 5 questions which were related to the characteristic of clean operating room had biger errors and difference.CONCLUSIONS The investigatees know about the basic requirements of the detecting method for bacterial concentration,but lack for the exact cognition of special requirements in clean operating room(e.g.State standard).The sticking point of the current problem is the misunderstanding on the 5 key questions.In order to make sure the excellent air condition in the cleaning operating room,the training of correlative specialty is very important.
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OBJECTIVE To investigate the effect of Hath1 on epithelial ridge (GER) cells from postnatal rat cochlear. METHODS An experimental method was developed which allowed the isolation and culture of GER cells from P1 rat cochleae using a combinatorial approach of enzymatic digestion and mechanical separation. The dissociated GER cells were cultured in DMEM+10% FBS. The GER explants infected with ad-Hath1 for 3 hours were cultured for 10 days and observed under scanning electron microscope. RESULTS The GER cell cultures tended to attach to the substratum and grow in patches which assume a polygonal morphology similar to that of epithelial cells in medium containing 10% serum. Stereociliary bundle-like structures were observed in the boundary of GER cell patches forced to express Hath1. CONCLUSION That GER cells which are likely hair cell progenitors could be cultured in vitro and generate stereociliary bundle-like structure when forced to express Hath1 suggests that the misexpression of Hath1 probably can induce the predifferentiation of pure hair cell progenitors into hair cells in vitro.
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0.05). A rise in CAP threshold and reduction in CM amplitude after perfusion were found in the other three groups(P
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Objective To observe the protective effect of ? receptor antagonist(Vasobral) against the aging process in rats otoconia morphology.Methods 15 healthy Wistar rats were subdivided into 3 groups:Normal control( n=5 ),aging group( n=5 ) and aging plus Vasobral group( n=5 ).Results The aged otlith took the form of clew-like fusion,flattening and some giant otoconia.And there was also some sphericand strange otoconia.In the region of transitional cells,there was many otoconia phagocytosis.Some scraps of saccule lost cilia,And the otoconia in the Vasobral group appeared to be spheric and some of them adhered together,but this phenomenon was not wide-spread.No giant otoconia was found in this group.The otoconia in the transitional region was less compared with the aged group.Conclusion ? receptor antagonist(Vasobral) has the effect of protection on the aging process of the otoconia.
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<p><b>OBJECTIVE</b>To investigate the prophylactic effect of low calcium concentration perilymph on noise-induced hearing loss.</p><p><b>METHODS</b>Forty guinea pigs with normal hearing weighing 250-350 g were assigned to five groups (8 in each group): (1) Ca(2+)-deficient perilymph perfusion (CDP) for 2 h; (2) white noise (120 dB SPL) exposure (WNE) only for 1 h, (3) combination of calcium-deficient perilymph perfusion and white noise (120 dB SPL) exposure (WNE + CDP); (4) normal artificial perilymph (NAP) perfusion for 2 h; and (5) white noise exposure + normal artificial perilymph perfusion (WNE + NAP) for 2 h. Compound action potentials (CAP) evoked by click was recorded from round window every 15 min. The cochleae from 5 animals in each group were examined with scanning electron microscope.</p><p><b>RESULTS</b>The CAP for group 1 experienced a threshold shift (TS) of 15-26 dB, while group 2 yielded a 46-59 dB TS and group 3 a 37-45 dB TS; no threshold shift occurred in group 4. The CAP TS in group 5 was 33-64 dB. The CAP TS of group 3 was less than that of group 2. After one hour of noise exposure, the CAP TS of group 3 were 45.92 +/- 2.90 dB and 59.30 +/- 3.95 dB in group 2. There were significant differences (P < 0.05) between groups 3 and 2. The CAP TS of group 3 was less than that of group 5 at the points of 1, 1.5 and 2 h after noise exposure. There was a significant difference between groups 3 and 5 (P < 0.01). Stereocilia of 89 OHC(3) were in disarray in five cochleae after noise exposure in group 2. The cuticular plates of 8 OHC(2),3 sank and the stereocilia became fused in only one animal cochlea after noise exposure in group 3 combined with low calcium perilymph perfusion.</p><p><b>CONCLUSIONS</b>Low calcium concentration appears to participate in preventing noise-induced hearing loss and the rising of calcium concentrations in inner hair cells after noise exposure, which may have been due to the opening of calcium channels in inner hair cells during noise exposure. The mechanism of the prophylactic effect might be caused by a lower calcium concentration in inner hair cells in the cochlea attenuating the influence of noise exposure on hearing loss; calcium deficient perilymph perfusion prevented calcium accumulation in inner hair cells of the cochlea. The motility of the OHCs might be partially inhibited by low calcium concentration that reduced noise-induced hearing loss in turn.</p>
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Animals , Action Potentials , Calcium , Physiology , Cochlea , Pathology , Physiology , Endolymph , Metabolism , Guinea Pigs , Hair Cells, Auditory , Metabolism , Hearing Loss, Noise-Induced , Perilymph , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the localization of Ca(2+)-ATPase (Ca(2+) pump) in the cochlear and its change after endolymphatic hydrops, and to study the relationship between compound action potential (CAP) threshold and the Ca(2+)-ATPase activety.</p><p><b>METHODS</b>The left endolymphatic sac was ablated to induce endolymphatic hydrops in fourteen healthy guinea pigs with normal action potential thresholds measured after a sliver ball electrode placed on the round window. Ca(2+)-ATPase activity was studied cytochemically using a lead citrate reaction in control and hydropic ears. The reaction product was lead phosphate particles as an expression of Ca(2+)-ATPase activity, observed with an eletron microscope.</p><p><b>RESULTS</b>Ca(2+)-ATPase activity is mainly found on the endolymphatic surface of Reis sner's membrane, the stereocilia and cuticular plate of inner and outer hair cells, and along the infolded plasma membrane of strial marginal cells. CAP thresholds of filtered click are increased and Ca(2+)-ATPase activity significantly decreased after endolymphatic hydrops in the mentioned locations.</p><p><b>CONCLUSIONS</b>CAP thresholds are increased and Ca(2+)-ATPase activity are significantly decreased in the cochlea after endolymphatic hydrops. These results suggest that there is a negative correlation between them.</p>
Subject(s)
Animals , Action Potentials , Calcium , Metabolism , Calcium-Transporting ATPases , Metabolism , Cochlea , Endolymphatic Hydrops , Guinea Pigs , Sodium , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of D-AP5 (D-2-amino-5-phosphonopentanoate, a specific NMDA-antagonist) on the increase of intracellular free Ca2+ concentration ([Ca2+]i) induced by glutamate in isolated cochlear inner hair cells (IHCs), and to detect the autoreceptors of the IHC membrane.</p><p><b>METHODS</b>When a laser scanning confocal microscope (LSCM) was used, the exogenous glutamate (Glu)-induced changes in [Ca2+]i of isolated IHCs and OHCs of guinea pig cochlea were observed with fluo-3, a fluorescent probe for [Ca2+]i. After D-AP5 or CNQX (6--cyano--7--nitroguinoxaline--2, 3--dione, a specific AMPA- antagonist) was administered, the exogenous glutamate (Glu)-induced changes in [Ca2+]i of isolated IHCs were recorded.</p><p><b>RESULTS</b>In the presence of a low concentration Glu (3.85 mumol/L), there was an increase of [Ca2+]i in IHCs, whereas there was no change in OHCs. When 50 mumol/L of D-AP5 was administrated in advance, Glu did not induce a corresponding increase in [Ca2+]i in IHCs, and 50 mumol/L of CNQX did not completely block the increase of [Ca2+]i in IHCs.</p><p><b>CONCLUSIONS</b>These results suggest that the autoreceptors existing in the IHC membrane are mainly of NMDA type, while there are relatively few AMPA receptors. Exogenous Glu is capable of increasing [Ca2+]i in IHCs by acting on the NMDA autoreceptor of IHCs in a positive feedback manner.</p>
Subject(s)
Animals , 2-Amino-5-phosphonovalerate , Pharmacology , 6-Cyano-7-nitroquinoxaline-2,3-dione , Pharmacology , Calcium , Metabolism , Excitatory Amino Acid Antagonists , Pharmacology , Glutamic Acid , Pharmacology , Guinea Pigs , Hair Cells, Auditory, Inner , Metabolism , In Vitro Techniques , Receptors, N-Methyl-D-AspartateABSTRACT
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.
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Objective To study the ultrastrueture of vestibular membrane of the cochlea in guineapig. Methods The vestibular membrane in guineapig's cochlea was observed under scanning electron microscope. Results The mesothelium on the perilymphatic side was formed by a single thin layer which was intermittently discontinuous. Microvilli were few on the surface of mesothelial cells. Nuclei of the cells were large and oval in shape. A single layer of polygonal epithelial cells were observed on the endolymphatical side. The irregular polygonal epithelial cells were round and oval in shape. Many microvilli were found on this side. The junction between these cells was very tight. Conclusion The vestibular membrane is a barrier between the scala media and scala vestibule and might play a role in ion and fluid homeostasis
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Objective To understand the structural characteristics and significance of the outer tunnel of corti's organ.Methods The structural characteristics of the outer tunnel were observed and analyzed with celloidin section and scanning electron microscope(SEM).Results The inner wall of outer tunnel consisted of the third row of outer hair cells(OHC) and the stalk of Deiter's cells in the basal and the first turn.Hensen's cells made up the outer wall of the outer tunnel.The stalks of Deiter's cells gradually moved to outer side to constitute the outer and top walls of outer tunnel from the basal turn to the top turn.The inner wall of outer tunnel consisted of the third OHC.The stalks of Deiter's cells moved to Hensen's cells to make the outer wall of the outer tunnel.The space of outer tunnel was gradually decreased.The outer tunnel was full of Deiter's cells in the third and fourth turn.The stalks of Deiter's cells tightly contacted with the Hensen's cells.Conclusion The structural characteristics of the outer tunnel play an important role in maintaining the stability of Corti's organ.
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Genome sequencing and bioinformatics is driving the discovery of drug targets and development of novel classes of broad spectrum antimicrobial compounds. The analysis of the complete genome sequences of microorganism available in the public databases offers the first insights into the drug targets discovery. This review discusses the use of genomic information in the finding of target genes for antimicrobial drug discovery.