Radiation-induced Injuries Of Middle Ear And It's Related Immunity Status Disorder In Relation With Different Micro Biota

PARTⅠ:Pathological observation of middle ear and immunity status difference in relation with different micro biotaObjective:To observe the difference of the immunity status and middle ear pathology between SPF rats and CV rats which were in different microbe levels.Methods:SPF and CV male SD rats,each were 8.To confirm oropharyngeal bacteria cultivation and determination;To observe the ultra structure of Eustachian tube mucosa with light microscope and scanning electron microscope;To detect peripheral blood T lymphocyte and its subtypes which were stained with fluorescence-labeled monoclonal antibodies by flow cytometry(FCM);to compare the spleen organ coefficients.Results:The bacteria cultivation ratio of CV rats is 100%and those of SPF rats is only 25%;The microbes differentiated from oropharynx of CV rats are Pseudomonas aerugnosa,Staphylococcus aureus,et al while the bacteria of SPF rats is only Proteus m irabilis.Light microscopy and electronic scanning microscopy show the ultra structure of Eustachian tube are normal and same,but light microscopy demonstrates the goblet cells on the surface of Eustachian tube of SPF rats are lesser than those of CV ones(P＜0.05);FCM showed the CD4+ T lymphocyte ratio of SPF rats are lesser than those of CV ones(P＜0.05);There is no statistical difference of spleen organ coefficients between two kinds of rats,but those of SPF rats are relatively small.Conclusions:There exist differences of oropharyngeal bacteria cultivation and determination between two kinds of rats.The goblet cell of Eustachian tube of CV rats suggests previous middle ear infection or chronic stimuli;FCM and spleen organ coefficients suggest that SPF rats are in the lower status to the stimulus of micro biota in the body. PARTⅡ:Radiation-induced injuries of otitis media and immunity status in relation with different micro biotaObjective:To observe differences of the radiation-induced injuries of Eustachian tube of middle ear and immune system between SPF rats and CV rats which were irradiated with gradient doses at nasopharyngeal region.To study the relationship of pathologically local and general immunity status of radiation-induced otitis media.Methods:SPF and CV male SD rats,each were 15.The nasopharyngeal fields of two kinds of rats were irradiated with 6MX linear accelerator at dose of 10Gy,20Gy,30Gy,5 each group,All were sacrificed 24-36hs after irradiation;To confirm oropharyngeal bacteria cultivation and determination;To observe the ultra-structural changes of Eustachian tube mucosa with scanning electron microscope;To detect the expression of intercellular adhesion molecule-1 of the Eustachian tube mucosa by SP immuno-histochemistry;To detect peripheral blood lymphocytes' surface markers(CD3+,CD4+,CD8+,CD54+) which were stained with fluorescence-labeled monoclonal antibodies by FCM.Results:The bacteria distinguished from CV rats are pathogen mostly and the one from SPF rats is still Proteus m irabilis uniquely.The scanning electronic microscopy shows under same irradiation dose the pathologically morphological injuries of CV Eustachian tubes are worse than those of SPF rats.ICAM-1 of Eustachian tube epithelium of SPF rats is lower than CV ones after radiation.The CD3+,CD4+ lymphocyte and the ratio of CD4+/CD8+ of CV rats decreased dramatically after radiation and in close relation with radiation doses while The CD8+ lymphocyte increased a bit.The CD3+,CD4+,CD8+ lymphocyte and the ratio of CD4+/CD8+ of SPF rats remain in a stable level.Statistic analysis shows there is correlation between ICAM-1 and T lymphocyte subtype changes after radiation.Conclusions:There exists the difference of radiation-induced injuries of otitis media and immune system in relation with different micro biota status.Micro-flora plays important roles in the radiation-induced normal tissue injuries and in the relationship between local and genenral immunity.