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目的在新西兰兔腰5椎体下终板种植结核分枝杆菌H37Rv标准菌株,构建兔脊柱结核模型。方法65只新西兰兔随机分为3组,实验组30只,对照组30只,空白对照组5只。弗氏完全佐剂致敏1月后,在腰5椎体下终板钻孔,实验组注入1×107CFU·m L~(-1)结核杆菌混悬液0.1m L,对照组注入等量生理盐水,空白对照组不作任何处理。术后观察兔一般情况,术后1个月、2个月对造模兔行X线、CT检查,术后2个月处死动物,行大体解剖观察,病椎取材作石蜡切片,对CT为阳性表现的兔椎旁肉芽组织进行结核杆菌培养。结果实验组25只、对照组28只,空白对照组5只兔完成实验。X线检查:术后1个月,实验组36.0%(9/25)的兔腰5、6椎体出现轻度骨质破坏;术后2个月,实验组52.0%(13/25)的兔腰5、6椎体骨质破坏较前加重,1只出现脊柱后凸畸形;对照组未见椎体骨质破坏,椎间隙无变化。CT扫描:术后1个月,实验组48.0%(12/25)的兔腰5、6椎体骨质破坏,病椎内细小死骨形成;术后2个月,实验组72.0%(18/25)的兔腰5、6椎体骨质破坏较前加重,5只椎旁脓肿形成;对照组未见椎体骨质破坏,无椎旁脓肿形成。术后2个月,实验组18只病椎石蜡切片苏木素-伊红(HE)染色显示有类上皮样细胞、结核坏死灶形成。结合影像学与组织病理学,术后2个月,兔脊柱结核造模成功率为72.0%(18/25)。18只兔的椎旁肉芽组织(CT阳性表现)结核杆菌培养阳性率为38.9%(7/18)。结论弗氏完全佐剂致敏的新西兰兔腰5椎体局部种植H37Rv标准菌株,可成功构建兔脊柱结核模型。
Objective To construct a standard model of Mycobacterium tuberculosis (H37Rv) in rabbits under the lumbar vertebrae 5 of New Zealand to construct a rabbit spinal tuberculosis model. Methods Sixty-five New Zealand white rabbits were randomly divided into three groups: experimental group (30), control group (30) and blank control group (5). Freund’s complete adjuvant sensitized 1 month after the end of lumbar vertebrae 5 vertebral drilling, the experimental group was injected with 1 × 107CFU · m L -1 Mycobacterium tuberculosis suspension 0.1m L, the control group was injected with the same amount Saline, blank control group without any treatment. Rabbits were observed postoperative general situation, 1 month after surgery, 2 months on the modeling of rabbit X-ray, CT examination, 2 months after the animals were sacrificed, the general anatomical observation, diseased vertebrae taken for paraffin section, CT was Positive rabbit paravertebral granulation tissue culture of Mycobacterium tuberculosis. Results Experimental group 25, control group 28, blank control group 5 rabbits to complete the experiment. X-ray examination: At 1 month after operation, 36.0% (9/25) of the rabbits in the experimental group had mild osteolysis on the lumbar and the 6th vertebra. At 2 months after operation, 52.0% (13/25) of the experimental group Rabbit lumbar 5,6 vertebral bone destruction worse than the previous, a kyphosis appeared; control group no vertebral bone destruction, no change in the intervertebral space. At 1 month after operation, 48.0% (12/25) rabbits in the experimental group suffered from osteolysis of the rabbit lumbar and 5,6 vertebrae, / 25) of rabbit lumbar 5,6 vertebral bone destruction worse than before, 5 paravertebral abscess formation; control group no vertebral bone destruction, no paravertebral abscess formation. At 2 months after operation, hematoxylin-eosin (HE) staining of 18 diseased paraffin sections of the experimental group showed epithelioid cells and tuberculous necrosis. Combined with imaging and histopathology, the successful rate of modeling of spinal tuberculosis in rabbits was 72.0% (18/25) 2 months after operation. The positive rate of mycobacterium tuberculosis culture in 18 rabbit paravertebral granulation tissue (CT positive) was 38.9% (7/18). Conclusions Freund’s complete adjuvant-sensitized New Zealand rabbit lumbar vertebrae 5 vertebrae are planted with H37Rv standard strains to successfully construct a rabbit spinal tuberculosis model.