超声弹性成像技术、P53基因与犬肿块性疾病的相关性研究

发布时间：2018-05-13 17:07

本文选题：超声 + 弹性成像　；参考：《河南农业大学》2015年硕士论文

【摘要】：为了提升宠物临床肿块性疾病的鉴别诊断水平,分别在超声影像学与分子生物学水平上对犬肿块性疾病进行应用研究。重点开展超声弹性成像技术、P53基因对宠物犬肿块性疾病鉴别诊断上的研究,并进一步探索上述两种新兴诊断技术在宠物临床的应用价值。本文以宠物犬为研究对象,以期为宠物临床肿块性疾病的鉴别诊断提供一定的依据和理论指导。试验选取宠物临床符合要求的犬肿块病例18例进行超声影像学研究,对获得的病例进行统计学分析,同时根据超声声像图采集方法和分析原则对18例肿块病例进行超声技术鉴别诊断,利用组织病理切片方法对18例肿块病例进行分析。实验基地饲养的雌性健康标准比格犬6只作为阴性对照,收集的宠物临床8例雌性犬恶性乳腺肿瘤样本作为阳性组,利用普通PCR对乳腺组织中P53基因进行半定量测定分析,利用实时荧光定量(real time Q-PCR)方法从RNA水平上定量测定P53基因的表达水平。宠物临床上犬肿块性疾病的发生在年龄、性别、品种以及发病部位上有着一定的规律可循。超声弹性成像系统中,应用超声弹性成像技术诊断肿块良恶性的准确率为94.4%(17/18)。单一根据弹性评分、应变比值、弹性面积比值三者中的一项诊断肿块良恶性与三者联合应用诊断肿块良恶性,结果比较应变比值(88.9%)与综合结果比较差异不显著(P=0.06,P0.05),弹性评分(66.7%)、弹性面积比值(72.2%)与综合结果比较差异极显著(P0.01)。应用传统超声技术(即二维灰阶超声、彩色血流多普勒超声)判定肿块良恶性的准确率分别为33.3%、55.5%,传统超声技术与超声弹性成像技术结果比较差异极显著(P0.01)。利用PCR测定结果表明,实验组与阴性对照组相比,实验组凝胶电泳成像基因条带的亮度与宽度明显较高;实时荧光定量PCR结果显示,阴性对照组中,P53基因与GAPDH基因的CT值结果均大于12.5,实验组中,P53基因与GAPDH基因的CT值结果均小于10;结果分析表明,恶性乳腺肿瘤组织中P53基因表达量,与正常乳腺组织中的P53基因表达量差异均显著,其中3号差异显著(P0.05),其余差异均极显著(P0.01)。结果表明:超声弹性成像技术在鉴别犬肿块良恶性方面与传统超声技术相比有着明显的特异性和准确性,该技术可作为今后宠物临床犬肿块良恶性的一种重要鉴别方法。P53基因表达水平可作为未来宠物临床监测犬乳腺肿块性疾病的一项重要分子水平指标。
[Abstract]:In order to improve the differential diagnosis of clinical mass diseases in pets, the application of ultrasound imaging and molecular biology were studied. The study on the differential diagnosis of pet dog mass disease with p53 gene was carried out and the application value of the above two new diagnostic techniques in pet clinic was further explored. The purpose of this study is to provide theoretical guidance for the differential diagnosis of pet clinical mass diseases. In this study, 18 cases of canine mass were selected for ultrasonic imaging study, and the obtained cases were statistically analyzed. At the same time, according to the ultrasonic image collection method and the principle of analysis, 18 cases of mass were diagnosed by ultrasonic technique, and 18 cases of mass were analyzed by histopathological section. Six female health standard Beagle dogs were used as negative control, and 8 female dog malignant breast tumor samples were collected as positive group. The p53 gene in mammary gland tissue was detected and analyzed by PCR. The expression of p53 gene was quantitatively measured at RNA level by real time Q-PCR method. The clinical occurrence of mass disease in pet has certain regularity in age, sex, variety and location. In the ultrasonic elastic imaging system, the accuracy of ultrasonic elastography in the diagnosis of benign and malignant tumors is 94. 4% or 17 / 18. A single diagnosis of benign and malignant masses was performed according to one of the three factors: elastic score, strain ratio and elastic area ratio. Results there was no significant difference between the strain ratio (88. 9%) and the comprehensive results (P 0. 06% P 0. 06% P 0. 05%, the elasticity score was 66. 7%, and the elastic area ratio was 72. 2%) compared with the comprehensive results, the difference was very significant (P 0. 01%). The accuracy of traditional ultrasound (i.e. two-dimensional gray-scale ultrasound, color flow Doppler ultrasound) in the diagnosis of benign and malignant tumors was 33.3and 55.5, respectively. The difference between traditional ultrasound technique and ultrasonic elastic imaging technique was very significant (P 0.01). The results of PCR analysis showed that the brightness and width of the gene bands in the experimental group were significantly higher than those in the negative control group, and the results of real-time fluorescence quantitative PCR showed that the gene bands in the experimental group were significantly higher than those in the negative control group. The CT value of p53 gene and GAPDH gene were higher than 12.5 in negative control group, and the CT value of p53 gene and GAPDH gene in experimental group were all less than 10. The results showed that the expression of p53 gene in malignant breast tumor was higher than that in control group. The expression of p53 gene was significantly different from that of normal mammary gland tissues, among which the difference of p53 gene expression was significant (P0.05) and the other differences were extremely significant (P0.01A). The results showed that the ultrasonic elastography was more specific and accurate than the traditional ultrasound in differentiating benign and malignant tumors in dogs. This technique can be used as an important method for the differential diagnosis of benign and malignant lesions in dogs. The expression level of p53 gene can be used as an important molecular level index for clinical monitoring of breast tumorous diseases in dogs in the future.
【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.292

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