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asbryobvrz · Wysłany: Pon 7:26, 14 Mar 2011

Experimental acute otitis media ear mucosa expression of heat shock protein

A methane hydrochloride (Tris-HCI) buffer (pH7.6), the temporary use of this buffer when added at 100:1 1O0 # g / ml phenylmethylsulfonyl fluoride (PMSF). Homogenate added to another plastic tube 10SDS5l to a final concentration of 0.1 (above operations carried out in an ice bath), a freezing and thawing 5 times 70 ℃, 15min / time, 11000r/min4 ℃ centrifuge 30min, take on serum a 70U refrigerator spare. Normal group with the law. See the documentation immune transfer method. Be. 2 Results 2.1 The performance of vaccinated animals, the spirit of the day a bit sluggish in guinea pigs, reduced activity, too, showed no abnormal urine and diet, nor was slow, swaying walk and so on. 1 day after inoculation to 7 days,[link widoczny dla zalogowanych], the guinea pig spirit, diet significantly decreased compared with the previous, not the individual activities of guinea pig, appears black ear,[link widoczny dla zalogowanych], ear canal discharging phenomenon. During the observation period, no treatment of guinea pigs, no guinea pigs died. 2.2 drawn on day 1 after inoculation seen listening to bubble slightly rough bone, bone open after dark, see the middle ear mucosa was congestion, edema, thickening of the middle ear cavity with purulent exudate, exudation As time increasingly thick liquid; tympanic membrane congestion, swelling, ulceration and even perforation. Thus, the guinea pig middle ear pneumoniae Infection Craig afraid, unloading of acute otitis media model was successfully JC] inOtorhinolaryn ~ o 【(China), AprU2000, Vol14, No42.3 general pathology from the middle ear 1 day after inoculation, the Ear infection manifested as severe submucosal edema. At the same time, the entire mucosal surface and have a lot of submucosa from polymorphonuclear cell infiltration, the thickness of the submucosa were significantly increased than normal. Accompanied by a large number of middle ear infection ear purulent exudate formation. 2.4 The heat shock response of middle ear mucosa under normal non-stress, guinea pig middle ear mucosa with only a lighter color, HSP70 expression (mean gray value of 368.) Middle ear infection suffered after stress, acute ear mucosa period significantly increased the expression of HSP70, was the strongest in the first 3 days (average gray value of 10230) while on day 3, 31kD occurring at the two bands, the first 5 days most (average gray value 15892),[link widoczny dla zalogowanych], to 7 days away. 17kD 1 to 5 days at both two bands, the highest on day 3 (average gray value of 25,426), reduced on day 7. 3 discuss the main observed experimental acute middle ear infection middle ear mucosa on the HSP70 response after vaccination and animal clinical manifestations and pathology of ordinary light microscope tips,[link widoczny dla zalogowanych], we have made an ideal animal model of otitis media, provides the basis for the study male guinea pigs were used to reduce confounding factors, results become more reliable. This data shows that in normal non-stress state, the guinea pig middle ear mucosa has a low level expression of HSP70, which is consistent with other reports Egusa. Middle ear infection stress, the guinea pigs in the expression of HSP70 at the same time, more strongly expressed HSP70 some common epitope of small molecular weight HSP. Egusa and other expression levels were significantly higher than normal non-stressed guinea pigs, but no low molecular weight HSP expression. suggest the same host of different products of bacterial heat shock response is different. These need to be further studied. middle ear infection by the bacteria occurred after the heat shock response . the one hand, middle ear mucosa has a protective effect on the {other hand, in certain circumstances, after Klebsiella pneumoniae infection of the host may have a common antigen or anti-HSP70 antibody,[link widoczny dla zalogowanych], the antibody may be caused by molecular simulation mechanism heat shock response of the middle ear mucosa appears immune injury. (This article is trapped 1 see inset on page 4)