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Wysłany: Sob 11:28, 12 Mar 2011
Temat postu: Inner ear ischemia on the expression of nitric oxi
Inner ear ischemia on nitric oxide synthase expression in the cochlea
School routine staining, light microscope photograph well. Cochlear N (~ nP, NA in situ hybridization: N0s oligo nucleotide primers from the edge of Microbiology, Chinese Academy of Sciences synthesis. Constructed in our lab NOSeDNA-PGEM-3ZF (+) plasmid, respectively, and touched by the E line nd Ⅲ digested nature of the DNA template, SP6RNA and T1RNA transcription polymerase synthesized complementary with the target mRNA sequence or the same anti, sense strand RNA, used to detect the target mRNA or control. whetstone prepared using random primers A probe just 0. cochlear frozen sections were pretreated with the D tag NO ~., P, NA of the antisense strand and sense strand probes miscellaneous change,
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, color, microscopy, photography. Bong Writing for Scientific Research Foundation of the Ministry of Health Author: 100083 Beijing Third Clinical Hospital,
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, Imperial College Medicine Department of Otolaryngology. basic research presentation. R) '. z. Northern blot: the same group of guinea pigs and 10 cochlear units to extract the total cochlear RNA. Each group were total RNA30, ug, by formaldehyde denaturing gel electrophoresis and capillary condensation chamber transfer film, with the P mark of cNOScDNA probe Noe, lzm blot hybridization, the scanned image points off quantitatively. Second, the results 1. Cochlea and the cerebellum NADPH-diaphorase staining: experimental group and control group 1,2 Go back inside the cochlea, outer hair cells (mc, OHC), spiral ganglion cells (sc.c) and marginal cells of stria vascularis, legs pulp shows positive blue reaction (Figure 1). Cytoplasm and processes of small neurons also positive blue reaction (Figure 2). 2. Cochlear NOSmO, NA in situ hybridization: e2, 1OS-eRNA antisense probe hybridization in the experimental group and control group 1,2 cochlea of the back IHC, OHC and the SC. C are seen in cytoplasm staining blue-purple granules,
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, staining was no significant difference in each group, each group was no positive staining of vascular pattern particles (circles 3-5). eNOSoRNA sense strand control hybridization probe, the experimental and control groups showed no positive staining of various parts of particles. 3. Northern blot hybridization: image analysis scan results. Experimental group and control group 1.2, the average absorbance () were 0.2433, O. 246O, and O. 2722, while the integral optical density (IOD), there were differences, 1993.0,2189.0 and 812.2, respectively. IOD asked the two experimental groups closer. Quite different with the control group. Third, the discussion of cerebral ischemia, stored in the presynaptic release of the Department of a large number of the acid, causing postsynaptic N-methyl-D-membrane aspartic acid over-excited by the piece, but excessive activation of NOS. Produce excessive NO. To its free radical activity while Emirates neurons poisoned J. Experimental study found that, NO involvement of glutamate in primary culture of cerebral cortex neurotoxicity, NOS inhibitors can prevent the N-methyl D-aspartate and related excitatory amino acid induced neurotoxicity, this effect can L arginine is reversed. NO not only to guide the boat to pull the Emirates nervous system excitatory amino acid caused by pieces of over-excited species by poisoning, in acute ischemic brains similar to cochlear ischemia can also occur when the excitatory amino acid receptors in neurotoxicity caused by over-excited. In which NO plays an important mediating role. N0 as a vascular relaxing factor in the inner ear shake cycle regulation plays an important role, has confirmed the existence of cochlear shake cycle 0s,
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, by vascular endothelial Taiwan into a release,
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, through activation of guanylate of the fermentation increased vascular smooth muscle cells cG (ring cGMP) levels, causing smooth muscle relaxation, resulting in vasodilatory effect. Involved in maintaining vascular bed resting tension and blood flow regulation. In addition, N0 can activate platelets through guanylate cyclase, increasing cGIVIP level, inhibition of platelet aggregation. Iheclddsh ~ e ~, etc. 【(] 994) were found to be given intravenously or 0s NOS competitive inhibitor L arginine natural substrate, it can reduce or increase the cochlear blood flow. (See this article from 1 to 5 loop insert page 31)
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