Oral lichen planus （OLP） is a chronic inflammatory disorder and premalignantlesion, of which the mechanisms are still obscure. In the present study, the expression levels of miR-96/182/183 cluster, miR-203, miR-375,...Oral lichen planus （OLP） is a chronic inflammatory disorder and premalignantlesion, of which the mechanisms are still obscure. In the present study, the expression levels of miR-96/182/183 cluster, miR-203, miR-375, and miR-769-5p in both tissues and exfoliative cells of OLP patients as well as healthy volunteers were detected, differentially expressed miRNAs were identified and their correlation with OLP was evaluated by a biplot method. Experimental results show that miR-203 is significantly up-regulated in patient lesion tissues in comparison to volunteer mucosa tissues. Moreover, the contra- dictory insignificant expression changes of miR-203 as well as miR-96/182/183 cluster in comparisons of exfoliative cell samples suggest that different cell compositions in OLP lesion have distinct miRNA regulation, which accords with the histological heterogeneity of OLP. Finally, biplot analyses indicate the expression of miR-203 and miR-96/182/183 cluster are positively correlated in patient lesions. These results provide miR-203 as a molecular indicator of heterogeneity of OLP, and also a potential diagnostic biomarker or therapeutic target that deserves further studies.展开更多
为了建立一种可以快捷、灵敏、安全地检测D型流感病毒(Influenza D virus,IDV)的实时荧光定量PCR(quantitative real time PCR,RT-qPCR)方法,试验首先将IDV NP基因作为检测目标,根据其序列的保守区设计了一对RT-qPCR引物,然后采用单一...为了建立一种可以快捷、灵敏、安全地检测D型流感病毒(Influenza D virus,IDV)的实时荧光定量PCR(quantitative real time PCR,RT-qPCR)方法,试验首先将IDV NP基因作为检测目标,根据其序列的保守区设计了一对RT-qPCR引物,然后采用单一变量法对RT-qPCR反应条件中的引物浓度和退火温度进行优化,建立了一种可检测IDV的RT-qPCR方法,并对该方法的灵敏度、重复性和特异性进行检测,最后应用该方法进行临床样本检测。结果表明:所建立的RT-qPCR方法的最佳引物浓度和退火温度分别为400 nmol/L和61℃,标准曲线为y=-3.572x+42.02(R^(2)=0.9993);该方法能检测到的最低拷贝数为8.30×10^(1)copies,灵敏度是常规PCR的100倍左右;组内重复和组间重复变异系数(CV)值均小于3%;同时检测牛呼吸道合胞体病毒(BRSV)、牛传染性鼻气管炎病毒(IBRV)、牛病毒性腹泻病毒(BVDV)、牛冠状病毒(BCoV)、牛副流感病毒3型(BPIV-3)、牛疱疹病毒4型(BHV-4)和牛流行热病毒(BEFV)的结果均为阴性;应用该方法对收集到的244份有呼吸道症状的牛鼻拭子样品进行检测,阳性率为1.23%。说明成功建立了IDV RT-qPCR检测方法,该方法具有灵敏度高、重复性和特异性好的特点,为IDV的防控提供了可靠的技术支撑。展开更多
基金National Natural Science Foundation of China(Grant No.91029711)
文摘Oral lichen planus （OLP） is a chronic inflammatory disorder and premalignantlesion, of which the mechanisms are still obscure. In the present study, the expression levels of miR-96/182/183 cluster, miR-203, miR-375, and miR-769-5p in both tissues and exfoliative cells of OLP patients as well as healthy volunteers were detected, differentially expressed miRNAs were identified and their correlation with OLP was evaluated by a biplot method. Experimental results show that miR-203 is significantly up-regulated in patient lesion tissues in comparison to volunteer mucosa tissues. Moreover, the contra- dictory insignificant expression changes of miR-203 as well as miR-96/182/183 cluster in comparisons of exfoliative cell samples suggest that different cell compositions in OLP lesion have distinct miRNA regulation, which accords with the histological heterogeneity of OLP. Finally, biplot analyses indicate the expression of miR-203 and miR-96/182/183 cluster are positively correlated in patient lesions. These results provide miR-203 as a molecular indicator of heterogeneity of OLP, and also a potential diagnostic biomarker or therapeutic target that deserves further studies.
文摘为了建立一种可以快捷、灵敏、安全地检测D型流感病毒(Influenza D virus,IDV)的实时荧光定量PCR(quantitative real time PCR,RT-qPCR)方法,试验首先将IDV NP基因作为检测目标,根据其序列的保守区设计了一对RT-qPCR引物,然后采用单一变量法对RT-qPCR反应条件中的引物浓度和退火温度进行优化,建立了一种可检测IDV的RT-qPCR方法,并对该方法的灵敏度、重复性和特异性进行检测,最后应用该方法进行临床样本检测。结果表明:所建立的RT-qPCR方法的最佳引物浓度和退火温度分别为400 nmol/L和61℃,标准曲线为y=-3.572x+42.02(R^(2)=0.9993);该方法能检测到的最低拷贝数为8.30×10^(1)copies,灵敏度是常规PCR的100倍左右;组内重复和组间重复变异系数(CV)值均小于3%;同时检测牛呼吸道合胞体病毒(BRSV)、牛传染性鼻气管炎病毒(IBRV)、牛病毒性腹泻病毒(BVDV)、牛冠状病毒(BCoV)、牛副流感病毒3型(BPIV-3)、牛疱疹病毒4型(BHV-4)和牛流行热病毒(BEFV)的结果均为阴性;应用该方法对收集到的244份有呼吸道症状的牛鼻拭子样品进行检测,阳性率为1.23%。说明成功建立了IDV RT-qPCR检测方法,该方法具有灵敏度高、重复性和特异性好的特点,为IDV的防控提供了可靠的技术支撑。
