Fusarium crown rot,mainly caused by Fusarium pseudograminearum,is a destructive disease in wheat production.To establish a rapid and reliable detection method for F.peasudeograminearum,the specific PCR primer pair(Fpg...Fusarium crown rot,mainly caused by Fusarium pseudograminearum,is a destructive disease in wheat production.To establish a rapid and reliable detection method for F.peasudeograminearum,the specific PCR primer pair(Fpg-F1;R2)was designed based on the RPB sequence,and real-time fluorescence quantitative PCR(qPCR)was used to validate the efficiency of the primer.The results showed that the primer pair had high specificity and sensitivity of 100 pg of DNA.Furthermore,the qPCR system for early and rapid detection of F.peasudeograminearum had an amplification efficiency of 87.5%and correlation coefficient of 0.99,and the pathologic threshold of F.pseudograminearum in soil was determined by using this detection system.It was found that F.pseudograminearum could cause Fusarium crown rot when the DNA concentration of F.pseudograminearum in field soil exceeded 213 pg·g^(-1).Hence,the qPCR-based method we developed for F.pseudograminearum detection has the advantages of high specificity and sensitivity,and can be used for rapid and early detection of F.pseudograminearum even in field soils.展开更多
VEGF （Vascular endothelial growth factor） signaling is critical for endothelial cell survival and maintenance of the vasculature. Deregulation of VEGF signaling contributes to the physiopathology of many diseases. H...VEGF （Vascular endothelial growth factor） signaling is critical for endothelial cell survival and maintenance of the vasculature. Deregulation of VEGF signaling contributes to the physiopathology of many diseases. However, the ways in which infection with Paracoccidioides brasiliensis affects VEGF signaling and the influence of immunization with rPb27 （recombinant protein Pb27） on VEGF signaling have not yet been studied. Animals were immunized with rPb27 and subsequently infected with a virulent strain of P. brasiliensis. The fungal load was evaluated by measuring CFU （colony-forming unit） and histology was performed to evaluate the inflammatory reaction. At the two time points analyzed, the PC （positive control） and TREAT （treated） animals had decreased levels of pulmonary VEGF compared to basal levels. However, in the immunized （Pb27） and treated mice （Pb27 ＋ TREAT）, VEGF expression remained unchanged after infection. In the case of VEGFR-2, the Pb27 and Pb27 ＋ TREAT groups showed increased levels of expression. Regarding the levels of the eNOS enzyme, only the Pb27 group did not reduce the expression levels relative to baseline. The immunization with rPb27 kept VEGF signaling, NO production and increased VEGFR-2 expression, after infection with P. brasiliensis. Thus, the authors infer that immunization with rPb27 protects mice from the disruption of VEGF signaling in Paracoccidioides brasiliensis infection.展开更多
文摘目的构建含rPB- DR启动子和TK自杀基因的重组复制缺陷型腺病毒并进行鉴定,为前列腺癌基因治疗的研究奠定基础。方法应用分子克隆技术,将rPB DR启动子连接至pBluescriptSK -TK上构建出pBluescriptSK -rPB- DR TK,再酶切出rPB -DR -TK目的片断,并将其连接至穿梭质粒pShuttle上。将构建好的pShuttle- rPB- DR -TK经PmeⅠ单切线性化后再以电穿孔法转化至E.coli.BJ5183AdEasy1内进行同源重组,抽提质粒经卡那抗性筛选及酶切鉴定正确的即为pAd- rPB- DR -TK,再经PacⅠ酶切回收后以脂质体法转染人胚肾293细胞,包装出完整腺病毒,然后大量扩增测定滴度,并行PCR鉴定。结果经酶切、PCR及测序鉴定证实成功构建了包含rPB -DR启动子和TK自杀基因的重组复制缺陷型腺病毒pAd rPB DR TK,包装、扩增后测病毒滴度为1.1×108TCID50/ml。结论成功构建出包含rPB DR启动子和TK自杀基因的重组复制缺陷型腺病毒并鉴定正确,可用于下一步前列腺癌基因治疗的研究中。
基金Yong Science and Technology Talent of AAAS(QNYC-201911)。
文摘Fusarium crown rot,mainly caused by Fusarium pseudograminearum,is a destructive disease in wheat production.To establish a rapid and reliable detection method for F.peasudeograminearum,the specific PCR primer pair(Fpg-F1;R2)was designed based on the RPB sequence,and real-time fluorescence quantitative PCR(qPCR)was used to validate the efficiency of the primer.The results showed that the primer pair had high specificity and sensitivity of 100 pg of DNA.Furthermore,the qPCR system for early and rapid detection of F.peasudeograminearum had an amplification efficiency of 87.5%and correlation coefficient of 0.99,and the pathologic threshold of F.pseudograminearum in soil was determined by using this detection system.It was found that F.pseudograminearum could cause Fusarium crown rot when the DNA concentration of F.pseudograminearum in field soil exceeded 213 pg·g^(-1).Hence,the qPCR-based method we developed for F.pseudograminearum detection has the advantages of high specificity and sensitivity,and can be used for rapid and early detection of F.pseudograminearum even in field soils.
文摘VEGF （Vascular endothelial growth factor） signaling is critical for endothelial cell survival and maintenance of the vasculature. Deregulation of VEGF signaling contributes to the physiopathology of many diseases. However, the ways in which infection with Paracoccidioides brasiliensis affects VEGF signaling and the influence of immunization with rPb27 （recombinant protein Pb27） on VEGF signaling have not yet been studied. Animals were immunized with rPb27 and subsequently infected with a virulent strain of P. brasiliensis. The fungal load was evaluated by measuring CFU （colony-forming unit） and histology was performed to evaluate the inflammatory reaction. At the two time points analyzed, the PC （positive control） and TREAT （treated） animals had decreased levels of pulmonary VEGF compared to basal levels. However, in the immunized （Pb27） and treated mice （Pb27 ＋ TREAT）, VEGF expression remained unchanged after infection. In the case of VEGFR-2, the Pb27 and Pb27 ＋ TREAT groups showed increased levels of expression. Regarding the levels of the eNOS enzyme, only the Pb27 group did not reduce the expression levels relative to baseline. The immunization with rPb27 kept VEGF signaling, NO production and increased VEGFR-2 expression, after infection with P. brasiliensis. Thus, the authors infer that immunization with rPb27 protects mice from the disruption of VEGF signaling in Paracoccidioides brasiliensis infection.
