Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located...Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located in ZIKV 5'UTR and virus production.Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining,Subsequently,liquid chromatographytandem mass spectrometry(LC-MS/MS),bioinformatics analysis,and Western blot were used to identify the candidate proteins binding to ZIKV 5'UTR.Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production,respecitvely.Results tRSA RNA pull-down assay,LC-MS/MS,and Western blot analysis showed that polypyrimidine tractbinding protein(PTB)bound to the ZIKV 5'UTR.Furthermore,dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV(t=10.220,P<0.001),while PTB knockdown had the opposite effect(t=4.897,P<0.01).Additionally,virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer(t=6.400,P<0.01),whereas reducing PTB expression level weakened virus infectivity(t=5.055,P<0.01).Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.展开更多
The Cyclonic-Static Microbubble Flotation Column (FCSMC) is currently a widely used, novel type of flotation device. The self-absorbing microbubble generator is the core component of this device. The structure of the ...The Cyclonic-Static Microbubble Flotation Column (FCSMC) is currently a widely used, novel type of flotation device. The self-absorbing microbubble generator is the core component of this device. The structure of the microbubble generator directly influences flotation column performance by affecting bubble size and distribution as well as gas holdup in the column. However, the complicated flow inside the generator results in high R&D costs and difficulty in testing. Thus, the CFD software, FLUENT, was used to simulate the gas-liquid two-phase flow inside a self-absorbing microbubble generator. The effect of area ratio, a key structural parameter, was studied in detail. Critical flow-field parameters including velocity, turbulent kinetic energy, minimum static pressure and gas holdup were obtained. The simulation results demonstrate that the optimum area ratio is 3.展开更多
Nowadays, machine learning is widely used in malware detection system as a core component. The machine learning algorithm is designed under the assumption that all datasets follow the same underlying data distribution...Nowadays, machine learning is widely used in malware detection system as a core component. The machine learning algorithm is designed under the assumption that all datasets follow the same underlying data distribution. But the real-world malware data distribution is not stable and changes with time. By exploiting the knowledge of the machine learning algorithm and malware data concept drift problem, we show a novel learning evasive botnet architecture and a stealthy and secure C&C mechanism. Based on the email communication channel, we construct a stealthy email-based P2 P-like botnet that exploit the excellent reputation of email servers and a huge amount of benign email communication in the same channel. The experiment results show horizontal correlation learning algorithm is difficult to separate malicious email traffic from normal email traffic based on the volume features and time-related features with enough confidence. We discuss the malware data concept drift and possible defense strategies.展开更多
文摘Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located in ZIKV 5'UTR and virus production.Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining,Subsequently,liquid chromatographytandem mass spectrometry(LC-MS/MS),bioinformatics analysis,and Western blot were used to identify the candidate proteins binding to ZIKV 5'UTR.Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production,respecitvely.Results tRSA RNA pull-down assay,LC-MS/MS,and Western blot analysis showed that polypyrimidine tractbinding protein(PTB)bound to the ZIKV 5'UTR.Furthermore,dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV(t=10.220,P<0.001),while PTB knockdown had the opposite effect(t=4.897,P<0.01).Additionally,virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer(t=6.400,P<0.01),whereas reducing PTB expression level weakened virus infectivity(t=5.055,P<0.01).Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.
基金Financial supports for this work provided by the National High Technology Research and Development Program of China (No.2008BAB31B02) is gratefully acknowledged
文摘The Cyclonic-Static Microbubble Flotation Column (FCSMC) is currently a widely used, novel type of flotation device. The self-absorbing microbubble generator is the core component of this device. The structure of the microbubble generator directly influences flotation column performance by affecting bubble size and distribution as well as gas holdup in the column. However, the complicated flow inside the generator results in high R&D costs and difficulty in testing. Thus, the CFD software, FLUENT, was used to simulate the gas-liquid two-phase flow inside a self-absorbing microbubble generator. The effect of area ratio, a key structural parameter, was studied in detail. Critical flow-field parameters including velocity, turbulent kinetic energy, minimum static pressure and gas holdup were obtained. The simulation results demonstrate that the optimum area ratio is 3.
基金the National Key Basic Research Program of China (Grant: 2013CB834204)the National Natural Science Foundation of China (Grant: 61300242, 61772291)+1 种基金the Tianjin Research Program of Application Foundation and Advanced Technology (Grant: 15JCQNJC41500, 17JCZDJC30500)the Open Project Foundation of Information Security Evaluation Center of Civil Aviation, Civil Aviation University of China (Grant: CAAC-ISECCA- 201701, CAAC-ISECCA-201702)
文摘Nowadays, machine learning is widely used in malware detection system as a core component. The machine learning algorithm is designed under the assumption that all datasets follow the same underlying data distribution. But the real-world malware data distribution is not stable and changes with time. By exploiting the knowledge of the machine learning algorithm and malware data concept drift problem, we show a novel learning evasive botnet architecture and a stealthy and secure C&C mechanism. Based on the email communication channel, we construct a stealthy email-based P2 P-like botnet that exploit the excellent reputation of email servers and a huge amount of benign email communication in the same channel. The experiment results show horizontal correlation learning algorithm is difficult to separate malicious email traffic from normal email traffic based on the volume features and time-related features with enough confidence. We discuss the malware data concept drift and possible defense strategies.
