本文選題：新型隱球菌　切入點：白念菌　出處：《第二軍醫(yī)大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:建立適合影像學(xué)研究使用的新西蘭大白兔的新型隱球菌、白念菌、煙曲霉菌肺炎動物模型,為進一步的臨床、病理、影像學(xué)診斷對照研究打下基礎(chǔ)。 材料與方法:選擇健康雄性新西蘭大白兔72只,分為新型隱球菌組、白念菌組、煙曲霉組,每組中設(shè)立實驗組18只,對照組6只,新型隱球菌對照組為未注射免疫抑制劑,而接種了新型隱球菌懸浮液,白念菌組和曲霉菌組對照組為注射了免疫抑制劑而注射了等量的生理鹽水。新型隱球菌實驗組和白念菌組使用相同的方法使兔達(dá)到免疫抑制狀態(tài):于實驗第1-5天,每日耳緣靜脈注射阿糖胞苷440mg/m~2,6天后隔日一次維持免疫抑制狀態(tài)。第4日起,所有兔給予萬古霉素15mg/kg、頭孢他定150mg/kg靜脈注射,每日1次;慶大霉素5mg/kg靜脈注射,隔日1次預(yù)防細(xì)菌感染。于第6日行經(jīng)皮氣管內(nèi)穿刺法分別接種配置好新型隱球菌0.4ml(濃度2.5×10~8個/ml)懸浮液和白念菌懸浮液0.2ml(濃度5×10~8個/ml)的懸浮液,白念菌對照注射等量的生理鹽水。煙曲霉組于實驗第1-5天、第8、9天,每日耳緣靜脈注射阿糖胞苷525mg/m~2、第1-2天每日耳緣靜脈注射甲基強地松龍5mg/kg,第4天起給予萬古霉素15mg/kg、頭孢他定150mg/kg靜脈注射,每日1次;慶大霉素5mg/kg靜脈注射,隔日1次預(yù)防細(xì)菌感染,于第2日行經(jīng)皮氣管內(nèi)穿刺法接種配置好的0.4ml煙曲霉懸浮液(濃度為1×10~8個/ml),對照組注射等量的生理鹽水。每組分別于實驗前和接種前抽耳緣靜脈血做血常規(guī)檢查了解粒細(xì)胞值的變化。每組所用兔在實驗開始時進行一次胸部CT檢查,接種后每隔一天行胸部CT檢查了解肺部的影像學(xué)表現(xiàn),并于接種后每2-3天隨機處死3-4只肺部有陽性表現(xiàn)兔(如有兔死亡量酌減),無菌操作下剖胸取出肺組織標(biāo)本行真菌培養(yǎng)和病理檢查,了解上述真菌在肺內(nèi)的生長和致病情況。 結(jié)果:1、三種組兔耳緣靜脈血粒細(xì)胞計數(shù)在注射免疫抑制劑后明顯降低,行配對t檢驗P＜0.05,有統(tǒng)計學(xué)意義。 2、新型隱球菌組的實驗組接種成功率為100%(18/18),模型建立成功率為83.33%(15/18)。對照組模型建立成功率為33.33%(2/6),與實驗組統(tǒng)計學(xué)有明顯差異。 3、白念菌組的實驗組接種成功率為100%(18/18),模型建立成功率為88.89%(16/18),對照組模型建立成功率為16.67%(1/6),與實驗組統(tǒng)計學(xué)有明顯差異。 4、煙曲霉組的實驗組接種成功率為100%(18/18),模型建立成功率88.89%(16/18),對照組接種和模型成功率均為0%。 5、胸部CT檢查主要異常表現(xiàn)為GGO、實變,可伴有支氣管血管束增粗、結(jié)節(jié)和網(wǎng)格線影。 結(jié)論:新西蘭大白兔免疫抑制后使用經(jīng)皮穿刺氣管內(nèi)接種法將新型隱球菌、白念菌和煙曲霉菌注入兔肺內(nèi)并進行定期CT掃描的方法了解肺內(nèi)病變情況可以建立適合影像學(xué)研究使用的上述三種真菌肺部感染的動物模型,此方法成功率高、具有可操作性、可重復(fù)性好,其中兔隱球菌肺炎模型的建立國內(nèi)外尚無文獻報道,為以后的影像病理對照研究打下了良好的基礎(chǔ)。
[Abstract]:Objective: to establish a new animal model of Cryptococcus neoformans, white fungus and Aspergillus fumigatus, which is suitable for imaging study, and lay a foundation for further clinical, pathological and imaging diagnosis.
Materials and methods: 72 male healthy New Zealand rabbits, divided into groups of Cryptococcus neoformans, Candida albicans, Aspergillus fumigatus group, set up the experimental group with 18 rats in each group, 6 rats in the control group, the control group is not Cryptococcus neoformans immune inhibitor, and inoculated with Cryptococcus albicans suspension. A group of bacteria and Aspergillus group control group injected immunosuppressant and injected with normal saline. Cryptococcus neoformans and Candida albicans were the experimental group using the same method to make rabbit immune suppression: on the 1-5 day, the daily auricular intravenous cytarabine 440mg/m~2,6 days after every other day for maintenance immunosuppression state. On the fourth day, all rabbits treated with vancomycin 15mg/kg, ceftazidime 150mg/kg intravenous injection, 1 times a day; gentamicin 5mg/kg intravenous injection, 1 every other day to prevent the bacterial infection. On the sixth day after percutaneous tracheal puncture inoculation respectively configured Cryptococcus 0.4ml (concentration of 2.5 * 10~8 /ml) suspension and Candida albicans suspension 0.2ml (concentration of 5 * 10~8 /ml) suspension of Candida bacteria injected with saline control group. Aspergillus fumigatus in the 1-5 days, 8,9 days, daily auricular intravenous cytarabine 525mg/m~2, the 1-2 day of ear vein injection of methylprednisolone 5mg/kg, fourth days for vancomycin 15mg/kg, ceftazidime 150mg/kg intravenous injection, 1 times a day; gentamicin 5mg/kg intravenous injection, 1 times a day to prevent bacterial infection, second underwent percutaneous tracheal puncture with a configured 0.4ml of Aspergillus fumigatus suspension (concentration of 1 * 10~8 /ml), the control group was injected with normal saline. Each group before experiment and inoculation before pumping ear vein blood for blood routine examination to understand the changes of neutrophils. The value of each rabbit at the start of a chest CT examination after inoculation Every other day to understand the chest CT examination of lung imaging performance, and in every 2-3 days after inoculation were sacrificed positive rabbits 3-4 lungs (such as rabbit mortality reduction), aseptic operation thoracotomy lung tissue specimens for fungal culture and pathological examination, understand the growth and pathogenicity of fungi in the lungs.
Results: 1, three groups of rabbit ear vein blood granulocyte count decreased significantly after injection of immunosuppressive agents, and paired t test was P < 0.05, with statistical significance.
2, the success rate of inoculation in Cryptococcus neoformans group was 100% (18/18), and the success rate of model establishment was 83.33% (15/18). The success rate of the control group was 33.33% (2/6), which was significantly different from that of the experimental group.
3, the inoculation success rate of the experimental group was 100% (18/18), the success rate of the model establishment was 88.89% (16/18), and the success rate of the control group was 16.67% (1/6), which was significantly different from that of the experimental group.
4, the successful rate of inoculation in the experimental group of Aspergillus fumigatus was 100% (18/18), the success rate of the model was 88.89% (16/18), and the success rate of inoculation and model in the control group was 0%.
5, the main abnormalities of the chest CT examination were GGO, real changes, accompanied by bronchovascular bundle thickening, nodules and grid lines.
Conclusion: New Zealand white rabbits were immunized after inhibition of using percutaneous intratracheal inoculation method of Cryptococcus neoformans, method of Candida albicans and Aspergillus fumigatus injected into rabbit lung and regular CT scan can be established for the understanding of pulmonary lesions imaging study using the above three kinds of pulmonary fungal infection animal model, the success rate of this method high maneuverability, good repeatability, has not been reported at home and abroad to establish rabbit model of cryptococcal pneumonia, imaging and pathologic for later control study to lay a good foundation.

【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R519;R-332

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