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目的评价诊断超声照射下微泡造影剂对绿荧光蛋白(green fluorescent protein,GFP)质粒在Wistar大鼠胫前肌中表达的作用。方法选用200 g左右成年Wistar大鼠40只,随机分为4组,每组10只,第1组只对大鼠胫前肌注射GFP质粒;第2组对大鼠胫前肌注射GFP质粒和造影剂的混合物;第3组对大鼠胫前肌注射GFP质粒和造影剂的混合物并加超声照射;第4组为阴性对照。12 d后处死动物,取胫前肌标本,制作冰冻切片后立即在荧光显微镜下观察。计数GFP阳性表达细胞数。部分组织经甲醛固定、石蜡包埋,光镜下观察病理改变。结果第3组GFP阳性表达细胞数最多[(345±19)个],与第1组[(109±15)个]、第2组[(111±14)个]差异有统计学意义(P<0.01)。第1组、第2组间差异无统计学意义(P=0.784)。所有受检组织光镜下检查未发现出血、坏死、炎症等病理改变。结论微泡造影剂在诊断超声照射下可以促进局部注射GFP质粒在大鼠胫前肌的转染。
Objective To evaluate the effect of microbubble contrast medium on the expression of green fluorescent protein (GFP) in the anterior tibial muscle of Wistar rats under ultrasound irradiation. Methods Forty adult Wistar rats weighing 200 g were randomly divided into 4 groups with 10 rats in each group. GFP plasmid was injected into the anterior tibial muscle of rats in group 1 only. GFP plasmid was injected into the anterior tibial muscle of rats in group 2 Group 3 was injected into the anterior tibial muscle of rats with a mixture of GFP plasmid and contrast medium and ultrasound irradiation; Group 4 was a negative control. After 12 days, the animals were sacrificed and the anterior tibial muscle samples were taken. Frozen sections were observed immediately under a fluorescence microscope. Count the number of GFP-positive cells. Some tissues were fixed by formaldehyde and embedded in paraffin, and pathological changes were observed under light microscope. Results The highest number of GFP positive cells in the third group [(345 ± 19)] was statistically significant compared with the first group [(109 ± 15)] and the second group [(111 ± 14) <0.01). There was no significant difference between group 1 and group 2 (P = 0.784). All subjects under light microscope examination found no bleeding, necrosis, inflammation and other pathological changes. Conclusion The microbubble contrast agent can promote the transfection of GFP plasmid injected into the anterior tibialis muscle in rats under the diagnostic ultrasound irradiation.