To develop a new high-throughput screening model for human high-density lipoprotein （HDL） receptor （CD36 and LIMPⅡ analogous-1, CLA-1） agonists using CLA-1-expressing insect cells. Methods With the total RNA of h...To develop a new high-throughput screening model for human high-density lipoprotein （HDL） receptor （CD36 and LIMPⅡ analogous-1, CLA-1） agonists using CLA-1-expressing insect cells. Methods With the total RNA of human hepatoma cells BEL-7402 as template, the complementary DNA （cDNA） of CLA-1 was amplified by reverse transcription-polymerase chain reaction （RT-PCR）. Bac-to-Bac baculovirus expression system was used to express CLA-1 in insect cells. CLA-1 cDNA was cloned downstream of polyhedrin promoter of Autographa californica nuclear polyhedrosis virus （AcNPV） into donor vector pFastBacl and recombinant pFastBacl-CLA-1 was transformed into E. coli DH10Bac to transpose CLA-1 cDNA to bacrnid DNA. Recombinant bacrnid-CLA-1 was transfected into Spodopterafrugiperda Sf9 insect cells to produce recombinant baculovirus particles. Recombinant CLA- 1 was expressed on the membrane of Sf9 cells infected with the recombinant baculoviruses. A series of parameters of DiI-lipoprotein binding assays of CLA-1-expressing Sf9 cells in 96-well plates were optimized. Results Western blot analysis and DiI-lipoprotein binding assays confirmed that CLA-1 expressed in insect cells had similar immunoreactivity and ligand binding activity as its native counterpart. A reliable and sensitive in vitro cell-based assay was established to assess the activity of CLA-1 and used to screen agonists from different sample libraries. Conclusion Human HDL receptor CLA-1 was successfully expressed in Sf9 insect cells and a novel high-throughput screening model for CLA-1 agonists was developed. Utilization of this model allows us to identify potent and selective CLA-1 agonists which might possibly be used as therapeutics for atherosclerosis.展开更多
Traffic classification research has been suffering from a trouble of collecting accurate samples with ground truth.A model named Traffic Labeller(TL) is proposed to solve this problem.TL system captures all user socke...Traffic classification research has been suffering from a trouble of collecting accurate samples with ground truth.A model named Traffic Labeller(TL) is proposed to solve this problem.TL system captures all user socket calls and their corresponding application process information in the user mode on a Windows host.Once a sending data call has been captured,its 5-tuple {source IP,destination IP,source port,destination port and transport layer protocol},associated with its application information,is sent to an intermediate NDIS driver in the kernel mode.Then the intermediate driver writes application type information on TOS field of the IP packets which match the 5-tuple.In this way,each IP packet sent from the Windows host carries their application information.Therefore,traffic samples collected on the network have been labelled with the accurate application information and can be used for training effective traffic classification models.展开更多
Uniqueness of our political systems and tremendous changes brought about by reform and opening up are reflected in language use,so contem porary Chinese political discourse has its own distinctiveness,and is especiall...Uniqueness of our political systems and tremendous changes brought about by reform and opening up are reflected in language use,so contem porary Chinese political discourse has its own distinctiveness,and is especially featured by increasing political neologisms,posing difficulties for interpreters and for China’s international publicity.Quality of political neologisms interpreting has a direct bearing on whether or not success of interpreting the whole discourse can be achieved.The authors believe that interpreting of contemporary Chinese political discourse,also for the purpose of com m unication can gain guidance from Bachman’s CLA Model.So the paper elaborates through exem plification on strategies for interpreting political neologisms in light of Bachm an’s Model,with a view to realizing discourse power of Chinese.展开更多
文摘To develop a new high-throughput screening model for human high-density lipoprotein （HDL） receptor （CD36 and LIMPⅡ analogous-1, CLA-1） agonists using CLA-1-expressing insect cells. Methods With the total RNA of human hepatoma cells BEL-7402 as template, the complementary DNA （cDNA） of CLA-1 was amplified by reverse transcription-polymerase chain reaction （RT-PCR）. Bac-to-Bac baculovirus expression system was used to express CLA-1 in insect cells. CLA-1 cDNA was cloned downstream of polyhedrin promoter of Autographa californica nuclear polyhedrosis virus （AcNPV） into donor vector pFastBacl and recombinant pFastBacl-CLA-1 was transformed into E. coli DH10Bac to transpose CLA-1 cDNA to bacrnid DNA. Recombinant bacrnid-CLA-1 was transfected into Spodopterafrugiperda Sf9 insect cells to produce recombinant baculovirus particles. Recombinant CLA- 1 was expressed on the membrane of Sf9 cells infected with the recombinant baculoviruses. A series of parameters of DiI-lipoprotein binding assays of CLA-1-expressing Sf9 cells in 96-well plates were optimized. Results Western blot analysis and DiI-lipoprotein binding assays confirmed that CLA-1 expressed in insect cells had similar immunoreactivity and ligand binding activity as its native counterpart. A reliable and sensitive in vitro cell-based assay was established to assess the activity of CLA-1 and used to screen agonists from different sample libraries. Conclusion Human HDL receptor CLA-1 was successfully expressed in Sf9 insect cells and a novel high-throughput screening model for CLA-1 agonists was developed. Utilization of this model allows us to identify potent and selective CLA-1 agonists which might possibly be used as therapeutics for atherosclerosis.
基金ACKNOWLEDGEMENT This research was partially supported by the National Basic Research Program of China （973 Program） under Grant No. 2011CB30- 2605 the National High Technology Research and Development Program of China （863 Pro- gram） under Grant No. 2012AA012502+3 种基金 the National Key Technology Research and Dev- elopment Program of China under Grant No. 2012BAH37B00 the Program for New Cen- tury Excellent Talents in University under Gr- ant No. NCET-10-0863 the National Natural Science Foundation of China under Grants No 61173078, No. 61203105, No. 61173079, No. 61070130, No. 60903176 and the Provincial Natural Science Foundation of Shandong under Grants No. ZR2012FM010, No. ZR2011FZ001, No. ZR2010FM047, No. ZR2010FQ028, No. ZR2012FQ016.
文摘Traffic classification research has been suffering from a trouble of collecting accurate samples with ground truth.A model named Traffic Labeller(TL) is proposed to solve this problem.TL system captures all user socket calls and their corresponding application process information in the user mode on a Windows host.Once a sending data call has been captured,its 5-tuple {source IP,destination IP,source port,destination port and transport layer protocol},associated with its application information,is sent to an intermediate NDIS driver in the kernel mode.Then the intermediate driver writes application type information on TOS field of the IP packets which match the 5-tuple.In this way,each IP packet sent from the Windows host carries their application information.Therefore,traffic samples collected on the network have been labelled with the accurate application information and can be used for training effective traffic classification models.
文摘Uniqueness of our political systems and tremendous changes brought about by reform and opening up are reflected in language use,so contem porary Chinese political discourse has its own distinctiveness,and is especially featured by increasing political neologisms,posing difficulties for interpreters and for China’s international publicity.Quality of political neologisms interpreting has a direct bearing on whether or not success of interpreting the whole discourse can be achieved.The authors believe that interpreting of contemporary Chinese political discourse,also for the purpose of com m unication can gain guidance from Bachman’s CLA Model.So the paper elaborates through exem plification on strategies for interpreting political neologisms in light of Bachm an’s Model,with a view to realizing discourse power of Chinese.
