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Micrium OS 开发者指南说明书

Micrium Tools Training: µC/Probe and SystemViewLabsMicrium’s µC/Probe and SEGGER’s SystemView can be invaluable for developers whose projects are based on the Micrium OS. The lab exercises described in this document will familiarize you with these two tools. The instructions for the lab assume that you have a Mighty Gecko Wireless Starter Kit (SLWSTK6000B), and that you’ve already insta lled µC/Probe and SystemView, as well as the Simplicity Studio IDE, on your PC. Download links for all of these tools are provided below.Download LinksSimplicity Studio: /products/development-tools/software/simplicity-studioThe Micrium OS example project prepared for these labs was developed using SimplicityStudio. You’ll need the IDE in ord er to build and run the project.µC/ProbeA visit to Micrium’s Web site to download µC/Probe is not necessary, since the tool was recently integrated into Simplicity Studio. However, you may need to use the IDE’s Package Manager to update your install and gain access to µC/Probe. Within the Package Manager, which you can find by clicking the button with the green, downward-facing arrow appearing near the upper-left corner of Simplicity Studio in the Launcher perspective, the link for installing µC/Probe is located on the Tools page, as indicated in the below screenshot. The Educational Edition of µC/Probe referenced by the link can be used without a license. If you’d like access to the full-featured Professional Edition of the tool, you can contact Micrium to get a one-year FAE license at no cost.SystemView: https:///downloads/free_tools/Setup_SystemView_V242.exe SystemView is a free tool from SEGGER. In order for the tool t o work properly, you’ll need the latest J-Link software, which is also available from SEGGER: https:///downloads/jlink/JLink_Windows_V614c.exe.Lab 1: Building and Running the Example ProjectIn this lab, you’ll build and run a Simplicity Studio example proj ect incorporating the Micrium OS. You’ll use this project in Lab 2 and Lab 3 to explore the features of µC/Probe and SystemView.Procedure:1.Your hardware platform for the labs is the Mighty Gecko Wireless Starter Kit. Youshould now connect one of your k it’s main boards (BRD4001A) to a radio board(BRD4161A). You should then establish a USB connection between your PC and thecombined boards. Your main board features a built-in J-Link debugger, and the USB connection will allow you to leverage this debugger within Simplicity Studio.2.If you’re not al ready running Simplicity Studio, you should start it now. You shouldmake sure that you’re in the Simplicity IDE perspective, which you can open by selecting Window>Perspective>Simplicity IDE.3.The example project for the labs is delivered in a zip file named Micrium-Tools-Lab.zip.You should now extract the contents of this zip file to a folder of your choice. To avoid any possible issues with long path names, it is recommended to keep the zip file contents relatively close to your root folder.4.You’ll now need to import the example project that was delivered in Micrium-Tools-Lab.zip. You should start the import process in Simplicity Studio by selecting File>Import…. On the dialog that subsequently appears, you should expand General and select Existing Projects into Workspace, as indicated in the below screenshot. You should then click the Next button.5.Within the second Import dialog, you should ensure that Select root directory is chosenand you should then click the corresponding Browse button. You must then navigate to the location of the project folder (named Tools-Lab-1) that was provided in Micrium-Tools-Lab.zip. Before clicking the Finish button, you should ensure that the Tools-Lab-1 is checked in the Projects field and that Copy projects into workspace is likewise checked, as indicated in the below screenshot.6.The example project should now appear in your Project Explorer, as indicated below.To clean any build artifacts that might exist for the project, you should right-click its name, Tools-Lab-1, and select Clean Project from the menu that appears. If the clean operation was successful, you should then attempt to build the project by again right-clicking the project name and this time selecting Build Project.7.The build operation for your project should have completed without any errors. (Thereshould have been one warning associated with deprecated interrupt code in the BSP.) In the case of a successful build, you should now attempt to download your project’s code to your board by right-clicking Tools-Lab-1in the Project Explorer and then selecting Debug As>1 Silicon Labs ARM Program.8.Simplicity Studio should automatically switch to the Debug perspective as a result ofyour actions in the previous step. Within this pers pective’s editor area, the tool should indicate that execution of your project is halted on the first line of the main() function. You should now start execution of the project’s code by clicking the Resume button shown below. LED0and LED1should then begin alternately blinking on yourboard.9.After confirming that your code is running and your LEDs are blinking, you can terminateyour debug session by right-clicking the Silicon Labs ARM MCU item that, as indicated inthe below screenshot, should appear in the Debug window. You should selectTerminate from the subsequent pop-up menu. Your LEDs should continue to blink afterthe debug session has ended, since your project’s code now resides in Flash memory.Lab 2: µC/ProbeIn this lab, you’ll use µC/Probe to visualize the internals of the project that you built in Lab 1. You’ll first monitor one of the project’s C variables, and you’ll then use µC/Probe’s kernel-awareness capabilities to view statistics output by the µC/OS 5 kernel on which the project is based.Procedure:1.You should now run µC/Probe. You can actually start the tool within Simplicity Studio byclicking the button shown in the below screenshot.2.µC/Probe makes it possible to view the values of an application’s variables as codeactually runs. In order to offer this capability, it needs information on the variables’ locations. It typically gets this information from an ELF file. When you built the Micrium OS example project following the procedure given in the previous section of this document, an ELF file, Tools-Lab-1.axf, should have been generated by Simplicity Studio and placed at the path listed below. In this path, <project_location> is the folder where your project resides. If you’re uncertain of this location, you should right-click the project’s name in Project Explorer and select Properties. If you then select Resource from the categories listed on the left side of the Properties dialog, you’ll be able to read your project folder from the Location field in the upper half of the dialog.<project_location>\GNU ARM v4.9.3 - Debug3.You can pass an ELF file to µC/Probe using the Symbol Browser located at the bottom ofthe tool’s main window. You should now click the Symbol Broswer’s ELF button, which is shown in the below screen shot. In the ensuing dialog, you should simply browse to the ELF file described above and then click the Open button.4.µC/Probe will be able to use your board’s built-in J-Link debugger to access variables inyour running example code. In order to establish J-Link as your preferred communication interface, you’ll need to select it in the Settings dialog. You should now open this dialog by clicking the Settings button shown below.5.As indicated in the below screenshot, the lower left corner of the Settings dialogcontains a list of Debug Interfaces containing just one item: J-Link. You should confirm that J-Link is selected, and you should then ensure that Don’t Change has been specified for the Interface Mode. Before clicking OK, you should choose Silicon Labs as the Manufacturer, and, using the provided table, select EFR32MG12PxxxF1024as your Device.6.µC/Probe provides a variety of graphical components that can be used for displaying thevalues of variables. You can access these components via the Toolbox located on the left side of the main program window. You should now instantiate a new component—a gauge—by first clicking Angular Gauges in the lower half of the Toolbox and thendragging and dropping the Semicircle 3gauge onto the data screen in the middle of µC/Probe’s program window. The data screen should then appear as shown in thebelow screenshot.7. A graphical component is not of much use until it has been associated with a variable, orsymbol. The Symbol Browser will be your means of associating a variable with your new gauge. If the Symbol Browser is not already displaying a list of C files corresponding to the code that you built in the first part of the lab, you can make such a list appear by expanding the entry for Tools-Lab-1.axf that appeared when you specified the path of this file in Step 2. Once the list is visible, you should expand the entry for the C file ex_main.c in order to make the variables contained in that file appear. You should then drag and drop the variable Ex_MainSpeed from the Symbol Browser to your gauge. As indicated below, the variable name will appear below thegauge to indicate that the two are associated.8.You should now click µC/Probe's Run button, which, as indicated below, is located in theupper left-hand corner of the tool's main program window. If you’re using the Educational Edition of the tool, you’ll be presented with a dialog indicating that you won’t have access to µC/Probe’s full set of features unless you upg rade to the Professional Edition. You can simply click this dialog’s Close button. You will then enter µC/Probe’s Run-Time mode. The dial on your gauge should begin moving from 0 to 100 and then back to 0. It should repeat this pattern indefinitely. Keep in mind that, with the Educational Edition, you will only be able to remain in Run-Time mode for one minute before a time-out occurs and another dialog prompting you to upgrade appears.9.µC/Probe can be used with nearly any embedded system—including those not based ona real-time kernel. However, the tool is especially helpful when paired with projectsthat incorporate Micrium’s embedded software modules, in part because of its kernel-awareness capabilities. These capabilities are implemented via a number of pre-populated data screens that show the values of different kernel variables. You should now stop µC/Probe (by clicking the Stop button located in the upper left-hand corner of the main program window) and add the kernel-awareness screens for µC/OS 5, the kernel featured in the example project, to your workspace. You can add the screens by first clicking Project1in µC/Probe's Workspace Explorer window and then clicking Screens>Micriµm OS Kernel (µC/OS-5), as indicated in the below screenshot. After making this change, you can again click Run to prompt µC/Probe to begin updating the new screens with kernel statistics.10. Within the main µC/OS 5 kernel-awareness screen, there are a number of subordinatescreens, including one for Task(s). As shown below, this screen displays the status of each task running in your project. You should now take a moment to look over the screen and familiarize yourself with the various tasks that the example incorporates.The percentage of CPU cycles consumed by the tasks is provided in the CPU Usage column, and you should note that, after the kernel’s Idle Task—which runs when no application tasks are ready—it is the Tick Task—the kernel task responsible for processing periodic tick interrupts—that consumes the most CPU cycles, with a usage of approximately 5.5%.11.After you’ve looked over Task(s), you should stop µC/Probe (via the Stop buttonmentioned in Step 9). You should then return to Simplicity Studio to make a simple change to the example project’s code. Within the IDE, you should open the file ex_main.c. You’ll be able to find this file by expanding the Micrium folder within the Project Explorer, and then similarly expanding Micrium_OS_V5.00 and examples. You should add the below two lines to ex_main.c, placing the new code at line 156, just below the variable declaration in the main() function. The result of your addition will be to change the frequency of tick interrupts received by the kernel, from 1 kHz to 100 Hz.OS_TASK_CFG Ex_MainCfgTick = {DEF_NULL, 256u, 4u, 100u};OS_ConfigureTickTask(&Ex_MainCfgTick);12.You should now build and run your modified project, following the same procedure thatyou used in Lab 1. Afterward, you should return to µC/Probe and again run your workspace. With tick interrupts now occurring at 1/10 of their original frequency, you should confirm that the overhead of the kernel’s Tick Task has experienced a similar decrease, meaning that the task’s CPU usage should be around 0.6%.Lab 3: SystemViewThis lab w ill walk you through the steps needed to use SEGGER’s SystemView with the example project featured in the previous two labs. With SystemView, you’ll be able to see the cont ext switches, interrupts, and kernel function calls that occur as the project runs.Procedure:1.Because the latest Micrium OS is a relatively new software module, it is not yetsupported by the version of SystemView available from the SEGGER Web site.Fortunately, though, it is fairly easy to update SystemView so that it will recognize applications based on MicriumOS. To make the update, you should now copy SYSVIEW_Micrium OS Kernel.txt, which was provided in Micrium-Tools-Lab.zip, and paste this file into the Description folder in the SystemView install path—Program Files (x86)\SEGGER\SystemView_V242.2.You should now run SystemView by clicking the tool’s entry in the Windows Start menu.3.When you run SystemView for the first time, you’ll be presented with the below dialog,asking whether you’d like to load a sample recording. You can simply click this dialog’s No button.4.In order to begin analyzing the behavior of your example project’s code, you’ll need torecord the code’s activity. To initiate recording, you should select Target>Start Recording within SystemView. You will then be presented with a Configuration dialog.Once you’ve verified that the contents of this dialog match the screenshot shown below, you should click the OK button to initiate recording.5.Shortly after you’ve started to record, you may be presented with the dia log shown inthe below screenshot. This dialog indicates that some of the data that would otherwise be displayed by SystemView was lost, because the buffer used to temporarily store that data on your target experienced overflows. You can simply click OK to close the dialog.For information on how to limit the potential for overflows, you can consult the last section in this document.6.Once you’ve gathered a few second’s worth of data, you should click the Stop Recordingbutton shown in the first of the two screenshots shown below. As the second screenshot indicates, the Timeline located near the center of the SystemView main program window should subsequently display all of the task and ISR activity that occurred during the period the recording was active. You now can begin investigating the execution of the example project’s code in detail.7.If you adjust the zoom on the timeline, you should be able to see, every 40 ms, thepattern of events depicted in the below screenshot. The example project incorporates three application tasks, in addition to a number of kernel, or system, tasks. One of the application’s tasks, labeled Post Task, peridocially performs a post operation on a semaphore, and that is what is shown in the screenshot. In the next couple of steps, you’ll adjust the period of the Post Task and then make an additional SystemViewrecording to confirm the results.8.The period of the Post Task is established by a variable in the same file that youmanipulated in Step 11 of the previous lab, ex_main.c. The variable is named Ex_MainDelayPostms and you have two different options for changing its value.One is to simply replace the variable’s initialization on line 371 of ex_main.c with thebelow code. You’l l then need to rebuild the example project and again download its code to your board. The second option involves µC/Probe. In addition to allowing you to read variables, µC/Probe offers a number of Writable Controls for changing variable values. You can drag and drop one of these controls—a Horizontal Slider, for example—into your workspace from Lab 2, and use the new control to adjust the value of ExMainDelayPostms.Ex_MainDelayPostms = 20;9.With the value of the variable adjusted, you should return to SystemView and make anew recording. Now the post operation described in Step 6 should be visible every 20 ms, as opposed to 40 ms. If time permits, you can make further adjustments to the code, changing, for example, the priority of the Post Task (established by the #define EX_MAIN_POST_TASK_PRIO), and you can use SystemView to observe the results of the changes.Limiting OverflowsThere are a few steps that you can take to limit overflows and maximize the amount of data recorded by SystemView. One of these is to increase the size of the buffer used to temporarily store SystemView data on your target before it is passed to the PC application. The buffer size is established by the #define SEGGER_SYSVIEW_RTT_BUFFER_SIZE in the file SEGGER_SYSVIEW_Conf.h. This file is contained in Micrium/Tools/SystemView/Config within the example project. The provided copy of SEGGER_SYSVIEW_Conf.h uses a buffer size of 4096, but you’re free to increase to any size that the hardware can accomodate.An additional measure, recommended by SEGGER for reducing the number of overflows, is to ensure that SystemView is the only tool using your J-Link. In other words, if you were previously running µC/Probe or the Simplicity Studio debugger while recording data, you should stop the other tools and try a new recording. With exclusive access to the J-Link, SystemView should have fewer impediments to capturing all of the example project’s events.。

Atg8, a ubiquitin-like protein required for autophagosome formation

Atg8,a Ubiquitin-like Protein Required for Autophagosome Formation,Mediates Membrane Tethering and HemifusionHitoshi Nakatogawa,1,2Yoshinobu Ichimura,1,3and Yoshinori Ohsumi1,*1Department of Cell Biology,National Institute for Basic Biology,Okazaki444-8585,Japan2PRESTO,Japan Science and Technology Agency,Saitama332-0012,Japan3Present address:Department of Biochemistry,Juntendo University School of Medicine,Bunkyo-ku,Tokyo113-8421,Japan. *Correspondence:yohsumi@nibb.ac.jpDOI10.1016/j.cell.2007.05.021SUMMARYAutophagy involves de novo formation of double membrane-bound structures called autophagosomes,which engulf material to be degraded in lytic compartments.Atg8is a ubiq-uitin-like protein required for this process in Saccharomyces cerevisiae that can be conju-gated to the lipid phosphatidylethanolamine by a ubiquitin-like system.Here,we show using an in vitro system that Atg8mediates the teth-ering and hemifusion of membranes,which are evoked by the lipidation of the protein and reversibly modulated by the deconjugation enzyme Atg4.Mutational analyses suggest that membrane tethering and hemifusion ob-served in vitro represent an authentic function of Atg8in autophagosome formation in vivo.In addition,electron microscopic analyses indicate that these functions of Atg8are in-volved in the expansion of autophagosomal membranes.Our results provide further insights into the mechanisms underlying the unique membrane dynamics of autophagy and also in-dicate the functional versatility of ubiquitin-like proteins.INTRODUCTIONAutophagy is an evolutionally conserved protein degrada-tion pathway in eukaryotes that is essential for cell survival under nutrient-limiting conditions(Levine and Klionsky, 2004).In addition,recent studies have revealed a wide variety of physiological roles for autophagy(Mizushima, 2005)as well as its relevance to diseases(Cuervo,2004). During autophagy,cup-shaped,single membrane-bound structures called isolation membranes appear and expand,which results in the sequestration of a portion of the cytosol and often organelles.Eventually,spherical, double membrane-bound structures called autophago-somes are formed(Baba et al.,1994),and then delivered to and fused with lysosomes or vacuoles to allow their contents to be degraded.Studies in S.cerevisiae have identiﬁed18ATG genes required for autophagosome formation,most of which are also found in higher eukary-otes(Levine and Klionsky,2004).Recent studies have shown that Atg proteins constituteﬁve functional groups: (i)the Atg1protein kinase complex,(ii)the Atg14-contain-ing phosphatidylinositol-3kinase complex,(iii)the Atg12-Atg5protein conjugation system,(iv)the Atg8lipid con-jugation system,and(v)the Atg9membrane protein recycling system(Yorimitsu and Klionsky,2005).The mechanisms by which these units act collaboratively with lipid molecules to form the autophagosomes,how-ever,are still poorly understood.Atg8is one of two ubiquitin-like proteins required for autophagosome formation(Mizushima et al.,1998;Ichi-mura et al.,2000).Because it has been shown that Atg8 and its homologs(LC3in mammals)localize on the isola-tion membranes and the autophagosomes,these proteins have been used in various studies as reliable markers for the induction and progression of autophagy(Kirisako et al.,1999;Kabeya et al.,2000;Yoshimoto et al.,2004). In S.cerevisiae,Atg8is synthesized with an arginine resi-due at the C terminus,which is immediately removed by the cysteine protease Atg4(Kirisako et al.,2000).The resulting Atg8G116protein has a glycine residue at the new C terminus and can serve as substrate in a ubiqui-tin-like conjugation reaction catalyzed by Atg7and Atg3, which correspond to the E1and E2enzymes of the ubiq-uitination system,respectively(Ichimura et al.,2000). Remarkably,unlike other ubiquitin-like conjugation sys-tems,Atg8is conjugated to the lipid phosphatidylethanol-amine(PE),thereby Atg8is anchored to membranes (Ichimura et al.,2000;Kirisako et al.,2000).Immunoelec-tron microscopy revealed that Atg8,probably as a PE-conjugated form(Atg8-PE),is predominantly localized on the isolation membranes rather than on the complete autophagosomes(Kirisako et al.,1999),suggesting that Atg8-PE plays a pivotal role in the process of autophago-some formation.The precise function of Atg8-PE,how-ever,has remained unknown.The conjugation of Atg8to PE is reversible;Atg4also functions as a deconjugation enzyme,resulting in the Cell130,165–178,July13,2007ª2007Elsevier Inc.165release of Atg8from the membrane(Kirisako et al.,2000). This reaction is thought to be important for the regulation of the function of Atg8and/or the recycling of Atg8after it has fulﬁlled its role in autophagosome formation.We reconstituted the Atg8-PE conjugation reaction in vitro with puriﬁed components(Ichimura et al.,2004). Here,we show using this system that Atg8mediates the tethering and hemifusion of liposomes in response to the conjugation with PE.These phenomena observed in vitro are suggested to reﬂect a bonaﬁde in vivo function of Atg8 in the expansion of the isolation membrane.Based on mutational analyses and structural information,the mech-anisms of Atg8-mediated membrane tethering and hemi-fusion as well as its regulation are discussed.This study sheds light on the molecular basis of unconventional membrane dynamics during autophagy,which is gov-erned by the Atg proteins.RESULTSLipidation of Atg8Causes Clustering of LiposomesIn VitroAs reported previously(Ichimura et al.,2004),when puri-ﬁed Atg8G116(hereafter,referred to as Atg8),Atg7,and Atg3were incubated with liposomes containing PE in the presence of ATP,Atg8-PE was efﬁciently formed (Figure1A,lanes1–6).Intriguingly,the reaction mixture became turbid during the incubation(Figure1B),which under a light microscope,was found to be a result of grad-ually forming aggregates(Figure1C).Both the degree of turbidity and the size of the aggregates appeared to corre-late with the amount of Atg8-PE produced in the mixture. Size-distribution analyses using dynamic light scattering (DLS)clearly showed that the aggregates formed in an Atg8-PE dose-dependent manner(Figure1D).These aggregates disappeared when the samples were treated with the detergent CHAPS(Figure1E,+CHAPS).In addi-tion,if a small amount of PE modiﬁed with theﬂuorescent dye7-nitro-2,1,3-benzoxadiazol-4-yl(NBD)was included in the liposome preparation,the aggregates became uniformlyﬂuorescent(Figure1E,NBD-PE).These results suggest that the aggregates generated during the produc-tion of Atg8-PE were clusters of liposomes.When the proteins were denatured with urea,the clus-ters of liposomes dissociated,although Atg8remained conjugated to PE(Figure1E,+urea and Figure1F,lane 2),indicating that the liposomes aggregated due to some function of the Atg8protein rather than an artifact caused by Atg8-PE as the lipid with the extraordinarily large head group.When the aggregates were sedimented by centrifugation,Atg8-PE co-precipitated with the lipo-somes(Figure1G,lane2),whereas Atg7,Atg3,and unconjugated Atg8did not(Figure1G,lane3).The sedimented liposomes containing Atg8-PE remained clustered even if they were brieﬂy sonicated(Figure1H, ppt.).These results suggested that Atg8-PE molecules function to tether together membranes to which they are anchored.Atg8-PE Also Mediates Liposome FusionWe also examined if membrane fusion occurred between the liposomes connected by Atg8-PE.To this end,we took advantage of a well-characterized lipid mixing assay (Struck et al.,1981).This method is based on energy transfer from NBD to lissamine rhodamine B(Rho),each of which is conjugated to PE.Because the amino group of the ethanolamine moiety is modiﬁed with the dyes, these lipids cannot be conjugated with Atg8.If both of the conjugated dyes are present at appropriate concen-trations in the same liposome,theﬂuorescence of NBD is effectively quenched by Rho(Figure2A,compare col-umns1and4).If a‘‘NBD+Rho’’liposome is fused with a‘‘nonlabeled’’liposome,which results in an increase of the average distance between the two dyes on the membrane,the NBDﬂuorescence will be dequenched.A mixture of the nonlabeled and NBD+Rho liposomes were subjected to the conjugation reaction.The resulting liposome clusters were dissociated by proteinase K treat-ment,followed byﬂuorescence measurements.Remark-ably,a signiﬁcant ATP-dependent increase of theﬂuores-cence was observed(ATP is required for the production of Atg8-PE;Figure2B,column6).This increasedﬂuores-cence was not observed with samples of nonlabeled lipo-somes alone,NBD+Rho liposomes alone,or a mixture of nonlabeled liposomes and liposomes containing NBD-PE but not Rho-PE(Figure2B,columns1-3).These results suggest that membrane fusion occurred between the lipo-somes tethered together by Atg8-PE.The increasedﬂuo-rescence was only observed if the reaction mixture was treated with proteinase K(Figure2B,columns4and6). This appeared to be due to the presence of Atg7and/or Atg3rather than Atg8or some effect of the clustering, because the NBDﬂuorescence was not increased by the addition of Atg4(Figure2B,column5),which detached Atg8from the membranes and dissociated the clusters of liposomes(see below).Instead,decreasing the con-centrations of the conjugation enzymes allowed the dequenching of the NBDﬂuorescence to be detected without proteinase K digestion(Figure2B,column7). The fusion of the liposomes was examined with various amounts of Atg8(Figure2C).The level of fusion increased Atg8dose-dependently and reached maximum at2m M (Figure2C).In contrast,a larger amount of Atg8produced an inhibitory effect(data not shown).This suggested that formation of the large aggregates resulted from excessive tethering by Atg8-PE,which no longer lead to fusion. We also carried out time-course experiments to roughly estimate the fusion rate using the lower concentrations of the conjugation enzymes(Figure2D),which eliminated the need for the proteinase K treatment(Figure2B).It should be noted that the incubation time includes the times re-quired for the formation of Atg8-PE and the subsequent tethering and fusion reactions.Under these conditions, the band of Atg8-PE could be seen on an SDS-PAGE gel after a10min incubation,and the reaction was completed within30min(Figure S1in the Supplemental Data available with this article online).It appeared that166Cell130,165–178,July13,2007ª2007Elsevier Inc.Figure1.Membrane Tethering Function of Atg8-PE In Vitro(A–C)Puriﬁed Atg8(10m M),Atg7(1m M),and Atg3(1m M)were incubated with liposomes(350m M lipids)composed of55mol%DOPE,30mol% POPC,and15mol%blPI in the presence(lanes1–6)or absence(lanes7–12)of1mM ATP at30 C for the indicated time periods,followed by urea-SDS-PAGE and CBB-staining(A),measurement of the absorbance at600nm(B),or observation under a light microscope(Nomarski images)(C).(D)Conjugation reactions with the various amounts of Atg8were performed as described in(A).After incubation for60min,the size distribution of the aggregates was examined using DLS measurements.d.nm,apparent diameter(nm).(E and F)The conjugation reactions were carried out as described in(A).They were further incubated at30 C for30min in the presence of either 6M urea or1%CHAPS and were then subjected to microscopy(E)or urea-SDS-PAGE and CBB-staining(F).The reaction was also performed with liposomes containing1mol%NBD-labeled DOPE(thus containing54mol%unlabeled DOPE),followed byﬂuorescence microscopy.Aﬂuo-rescence image with aﬁlter for YFP(NBD-PE,FL)and a Nomarski image(NBD-PE,DIC)are shown.(G and H)Atg8(30m M),Atg7(2m M),and Atg3(2m M)were incubated with liposomes(350m M lipids)consisting of70mol%DOPE and30mol% POPC in the presence of1mM ATP at30 C for45min(total).The mixture was microcentrifuged at15,000rpm for10min to generate the pellet (ppt.)and the supernatant(sup.)fractions.The fractions were brieﬂy sonicated and were analyzed by urea-SDS-PAGE(G)or observed under a light microscope(H).In this experiment,blPI was omitted to prevent Atg7and Atg3from tightly binding to the liposome.We showed that Atg8could also cause hemifusion of liposomes with this lipid composition.Cell130,165–178,July13,2007ª2007Elsevier Inc.167the liposomes began to fuse shortly after the formation of Atg8-PE.The fusion reaction proceeded concurrently with the conjugation reaction and continued for 30min after the completion of the Atg8-PE production (Figure 2D,ﬁlled circles).Small liposomes <100nm in diameter tend to sponta-neously fuse (Chen et al.,2006),and the liposomes we used in the above experiments were 70nm in diameter (Figure 1D).However,we also showed that Atg8-PEcaused a signiﬁcant level of fusion between larger lipo-somes in spite of their stability against spontaneous fusion (Figure S1).Taken together,these results suggest that not only tethering but also fusion of the liposomes is mediated by Atg8-PE.The Atg8-Mediated Membrane Fusion Is Hemifusion Recent in vitro studies on membrane fusion mediated by SNARE proteins and a class of viral proteinsrevealedFigure 2.Membrane Hemifusion Occurs between Liposomes Tethered by Atg8-PE(A and B)Nonlabeled (55mol%DOPE,30mol%POPC,and 15mol%blPI),NBD-labeled (55mol%DOPE,29mol%POPC,15mol%blPI,and 1mol%NBD-DOPE),and NBD+Rho-labeled (55mol%DOPE,27.5mol%POPC,15mol%blPI,1mol%NBD-DOPE,and 1.5mol%Rho-DOPE)liposomes were mixed in the differ-ent combinations and ratios indicated.Their relative intensities of the NBD ﬂuorescence ob-served are shown (the value obtained with a 4:1mixture of the nonlabeled and NBD+Rho lipo-somes was deﬁned as 1)(A).These mixtures of liposomes were incubated with Atg8(4m M),Atg7(0.5or 1.0m M),and Atg3(0.5or 1.0m M)in the presence (ﬁlled columns)or absence (open columns)of 1mM ATP for 60min,and were then treated with 1unit/ml apyrase.The mixtures were further incubated for 30min with the buffer (columns 4and 7),1m M Atg4(columns 5and 8),or 0.2mg/ml proteinase K (columns 1-3,6and 9),followed by measure-ment of the NBD ﬂuorescence.The experi-ments were repeated three times and the average ﬂuorescence values divided by those obtained from the original liposome samples (F/F 0)are presented with error bars for the stan-dard deviations (B).(C)A 4:1mixture of the nonlabeled and NBD+Rho liposomes was incubated with various amounts of Atg8,1.0m M Atg7,and 1.0m M Atg3in the presence (open circles)or absence (ﬁlled circles)of ATP,and the samples were then treated with proteinase K,followed by measuring the NBD ﬂuorescence.(D)The conjugation reactions were performed with the mixed liposomes used for the lipid mixing assay,0.5m M Atg7,and 0.5m M Atg3in the presence or absence of Atg8(4m M)and ATP.After incubation for the indicated time periods,an aliquot of the samples was immediately subjected to the ﬂuorescence measurements.The values that were obtained by subtracting the signals observed in the absence of ATP from those observed in the presence of ATP are presented.(E)The lipid mixing assay was performed with 4m M Atg8,1m M Atg7,and 1m M Atg3in the presence or absence of ATP (white bars in columns 3and 2,respectively)as described in(C).For PEG-induced fusion reactions,the mixed liposomes were incubated at 37C for 30min in the presence or absence of 12.5%PEG 3350(white bars in columns 5and 4,respectively).These samples as well as the original liposomes (column 1)were then incubated with 20mM sodium dithionite on ice for 20min in the presence (black bars)or absence (gray bars)of 0.5%Triton X-100,followed by the NBD ﬂuorescence measurement.168Cell 130,165–178,July 13,2007ª2007Elsevier Inc.that fusion proceeds through an intermediate state called hemifusion,in which outer(contacting)leaﬂets of two apposed lipid bilayers merge,while inner(distal)leaﬂets remain intact(Chernomordik and Kozlov,2005).It was also reported that fusion can be arrested or delayed at the hemifusion state under some conditions.Therefore, we investigated whether the liposome fusion caused by Atg8in vitro was complete fusion(the merger of both inner and outer leaﬂets)or hemifusion(Figure2E).This can be examined using the membrane impermeable reductant sodium dithionite that selectively abolishes theﬂuores-cence of NBD conjugated to the lipid head group in the outer leaﬂet(Meers et al.,2000).Accordingly,when so-dium dithionite was added to the original liposomes,the background level of the NBDﬂuorescence was decreased by about50%,whereas it was hardly detected in the pres-ence of the detergent(Figure2E,column1).Strikingly,the NBDﬂuorescence increased by the Atg8-mediated fusion was totally eliminated by addition of sodium dithionite to the same level as those observed in the original lipo-somes and the reaction mixture incubated without ATP (Figure2F,columns1-3).Whereas,we conﬁrmed that in liposome fusion induced by polyethylene glycol(PEG), which causes complete fusion(Akiyama and Ito,2003), about half of the increasedﬂuorescence was retained af-ter sodium dithionite treatment(Figure2F,columns4and 5).Taken together,membrane fusion mediated by Atg8 in vitro was suggested to be hemifusion.To obtain direct evidence of hemifusion,we analyzed the morphology of liposomes by electron microscopy (Figures3A–3E),in which liposomal membranes were observed as double white lines that correspond to the outer and inner leaﬂets.When the clusters of liposomes formed by Atg8-PE were analyzed,tight junctions between the liposomes were observed(Figures3B and 3C,arrowheads).Consistent with the biochemical results suggesting that complete fusion does not occur,the size of the individual liposomes did not appear to signiﬁcantly increase(compare Figures3A and3B).Instead,hallmarks of hemifusion,trifurcated structures formed by one contin-uous outer leaﬂet and two separate inner leaﬂets,could be observed at the junction between the liposomes(Figures 3C–3E,arrows).These results strongly support our con-clusion that Atg8-PE causes hemifusion of liposomes. Atg8Forms a Multimer in Responseto the Conjugation with PEWe also performed immunoelectron microscopy of the liposomes clustered by Atg8-PE(Figures3F–3I).Intrigu-ingly,Atg8-PE tended to be enriched at the junction be-tween the liposomes(Figures3G–3J).While,if the mixture incubated without ATP was similarly analyzed,the signal was rarely observed on the liposome(Figure3F;the gold particles observed should represent unconjugated Atg8 adsorbed onto the grid).These results indicate that Atg8-PE is directly involved in the tethering and hemifu-sion of liposomes.We observed that‘‘naked’’liposomes do not associate with liposomes carrying Atg8-PE(data not shown), suggesting that tethering should be achieved due to interactions between Atg8-PE molecules on different membranes.We therefore examined the intermolecular interaction of Atg8-PE by crosslinking experiments(Fig-ure4).The reaction mixture containing Atg8-PE or unconjugated Atg8was incubated with the lysine-to-lysine reactive crosslinker DSS.We found that a crosslink adduct with a molecular weight of 24kDa on a SDS-PAGE gel speciﬁcally appeared in the sample containing Atg8-PE(Figure4A,lane5).Considering the molecular weights of the proteins included,this adduct should represent an Atg8-PE homodimer.Immunoblotting anal-yses with anti-Atg8revealed that two additional crosslink adducts of about37and100kDa were also speciﬁcally produced in the Atg8-PE-containing sample(Figure4B, lanes2–4and Figure S2).These products were immuno-stained neither with anti-Atg7nor anti-Atg3(data not shown),suggesting that they represent a trimer and a larger multimer of Atg8-PE,respectively,and thus that Atg8multimerizes in response to PE conjugation. We also showed that this multimerization correlates with the membrane tethering ability of Atg8(see below), indicating that interactions between Atg8-PE molecules on different membranes are responsible for the tethering of the membranes.The Membrane-Tethering and HemifusionFunctions of Atg8Are Modulatedby the Deconjugation Enzyme Atg4Our results suggest that the membrane-tethering and hemifusion functions of Atg8are evoked by the conjuga-tion with PE,whereas Atg4functions as a deconjugase that cleaves the linkage between Atg8and PE(Kirisako et al.,2000).We reconstituted this reaction in vitro.After producing Atg8-PE using the conjugation reaction,the re-action was terminated by adding apyrase to deplete the remaining ATP.When puriﬁed Atg4was then added, Atg8-PE was rapidly and almost completely deconjugated (Figure4C,lanes1-6).In contrast,when the Atg4was pretreated with the cysteine protease inhibitor N-ethylma-leimide(NEM),the deconjugation reaction did not occur (Figure4C,lanes7–12).These results clearly show that Atg4is sufﬁcient for the deconjugation of Atg8-PE.Upon deconjugation,the liposome aggregates immediately dissociated(Figure4D).In addition,we found that multi-merization of Atg8is also reversible;the crosslink adducts corresponding to the Atg8-PE dimer(Figure4A,lane6)as well as the trimer and the multimer(data not shown)were hardly formed when DSS was added after the deconjuga-tion reaction.We also showed that the presence of Atg4in the conjugation reaction retarded the accumulation of Atg8-PE and accordingly interfered with the tethering and hemifusion of the liposomes(data not shown).It was indicated that membrane tethering and hemifusion by Atg8can be regulated by the balance between the conjugation and deconjugation reactions.Cell130,165–178,July13,2007ª2007Elsevier Inc.169Identiﬁcation of Mutations that Impair the Postconjugational Function of Atg8In VivoIf the function of Atg8-PE we observed in vitro was involved in autophagosome formation in vivo,Atg8mutants deﬁ-cient for this function should result in defective autophagy.To examine this idea,we performed structure-based and systematic mutational analyses of Atg8(Figure 5).The structures of mammalian homologs revealed that Atg8family proteins consist of two domains:an N-terminal heli-cal domain (NHD)and a C-terminal ubiquitin-like domain (ULD)(Paz et al.,2000;Coyle et al.,2002;Sugawara et al.,2004;Figures 5E–5H).Among the highly conserved residues in the ULD,we selected those with side chains that were exposed on the domain surface (Figure 5A),and individually replaced them with alanine,except that serine was substituted for Ala75.Consequently,we didnot mutate residues suggested to be important for interac-tions with the conjugation enzymes,because these resi-dues are conserved only for their hydrophobic nature (Sugawara et al.,2004).The Atg8variants were expressed from centromeric plasmids in D atg8yeast cells,and their autophagic activities were biochemically assessed (see Supplemental Experimental Procedures ).In nutrient-rich media,the autophagic activity was low in all of the mutant cells as well as in the wild-type cells (data not shown).In contrast,in nitrogen starvation conditions,which strongly induced autophagy,a number of mutants were found to have defective autophagic phenotypes (Figure 5B).Ala-nine replacement of seven residues,Ile32,Lys48,Leu50,Arg65,Asp102,Phe104,and Tyr106,signiﬁcantly impaired the autophagic activity to 30%–60%of that of the wild-type (Figure 5B).Immunoblotting analyses showedthatFigure 3.Electron Microscopic Analyses of the Liposomes Tethered and Hemi-fused by Atg8-PEConjugation reactions were performed with 4m M Atg8,1m M Atg7,and 1m M Atg3in the presence (B–E and G–I)or absence (A and F)of ATP for 60min and subjected to phospho-tungstic acid-staining and electron micros-copy (A–E).The junctions between the lipo-somes and the structures suggested to represent hemifusion are indicated with arrow-heads and arrows,respectively.The same samples were also subjected to immunostain-ing using puriﬁed anti-Atg8-IN-13and anti-rab-bit IgG conjugated with 5nm gold particles,fol-lowed by phosphotungstic acid-staining and electron microscopic observation (F–I).To as-sess the enrichment of Atg8-PE at the junction of the liposomes (J),images of two contacting liposomes as shown in G and H were randomly picked up (n =41).The lengths of contacting (CR)and noncontacting regions (non-CR)of the liposomal membranes were measured (white bars),thereby the number of gold parti-cles on each region (gray bars)was divided,in which the length of the contacting region was doubled,to calculate the linear density (black bars).The average values are presented with error bars for the standard deviations.170Cell 130,165–178,July 13,2007ª2007Elsevier Inc.a substantial amount of each of the Atg8mutant proteins accumulated in the cells (Figure 5D),although there were some differences in their mobilities in SDS-PAGE analysis;for instance the PE-conjugated and unconjugated forms of the D102A mutant exhibited almost the same mobility.None of the mutations signiﬁcantly affected the formation of Atg8-PE (Figure 5D),suggesting that the mutations impaired a function of Atg8that was exerted after the conjugation with PE.Notably,these mutants accumulated different levels of unconjugated Atg8under the starvation conditions (Figure 5D,starvation),which allowed us to classify them into three groups.For the class I mutants K48A and L50A,the levels of the unconjugated forms were similar to that of the wild-type (Figure 5D,denoted in purple).On the other hand,compared to the wild-type,lower levels of the unconjugated forms were detected in the class II mutants I32A,D102A,F104A and Y106A (Figure 5D,denoted in red),whereas a larger amount of the unconju-gated class III mutant R65A accumulated (Figure 5D,denoted in orange).We then mapped the mutated resi-dues onto the three-dimensional structure of LC3(Suga-wara et al.,2004),which revealed that class of the mutant corresponded to the location of the mutation.All the class II residues were clustered in a speciﬁc region on the ULD (hereafter,referred to as the class II region),and the two neighboring class I residues were located close to the class II region (Figure 5E).In contrast,the class III residue was located away from the other mutated residues (Fig-ures 5G and 5H).The NHD of Atg8contains two helices:a 1and a 2(Figure 5A).We constructed two mutants,one with a dele-tion of a 1(D N8)and a second bearing deletions of both helices (D N24).It was shown that the NHD is involved in autophagy partially but signiﬁcantly;the D N8and D N24mutations decreased the autophagic activity by about 30and 40%,respectively (Figure 5C).We also showed that the deletions did not affect the stability of the proteins or the formation of the PE conjugates (Figure S3).Effects of the Atg8Mutations on the Membrane-Tethering FunctionWe next examined whether the mutations affected the liposome-clustering ability of Atg8in vitro (Figure 6).Figure 4.The Membrane-Tethering Function and Multimerization of Atg8Are Reversibly Regulated in Response to Conjugation with PE(A)Conjugation reactions were performed as described in Figure 3in the presence (lanes 2,3,5,and 6)or absence (lanes 1and 4)of ATP.They were mixed with 1unit/ml apyrase,and then incubated with (lanes 3and 6)or with-out (lanes 1,2,4,and 5)puriﬁed Atg4(0.5m M)at 30 C for 30min.These samples were further incubated with (lanes 4–6)or without (lanes 1–3)100m M DSS for 30min,and then analyzed by urea-SDS-PAGE and CBB-staining.(B)The reaction mixture including ATP was in-cubated with different concentrations of DSS as indicated,followed by urea-SDS-PAGE and immunoblotting with anti-Atg8-IN13.We also identiﬁed a crosslink product that reacted with anti-Atg3(Atg8xAtg3).(C and D)The conjugation reactions performed as described in Figure 1A were mixed with 1unit/ml apyrase.Atg4(0.5m M)pretreated with (lanes 7–12)or without (lanes 1–6)10mM NEM was then added,and the samples were incubated for the indicated time periods and subjected to urea-SDS-PAGE and CBB-stain-ing (C).The same samples were also observed under a light microscope (D).Cell 130,165–178,July 13,2007ª2007Elsevier Inc.171。

英语b级考试题

英语b级考试题英语B级考试题是许多英语学习者为提高英语水平而重点准备的考试之一。

这个考试对于英语学习者来说有着重要的意义，它不仅可以检测学生的英语水平，还可以帮助他们提高英语听、说、读、写各方面的能力。

下面是一个关于英语B级考试的综合练习题，请认真完成：Part I Listening Comprehension （听力理解）Section A （第一部分听力理解）1. What time will the lecture start?A. 9:00.B. 9:15.C. 9:30.2. Where is the man going to be next Monday?A. At the office.B. At the airport.C. At the hotel.3. What is the woman’s job?A. She is a teacher.B. She is a doctor.C. She is a nurse.4. What does the man suggest the woman do?A. Take up swimming.B. Read more books.C. Listen to music.5. Why did the man visit the woman?A. To pick up a book.B. To return a book.C. To buy a book.Section B （第二部分听力理解）听下面一段短对话，回答第6和第7两个小题。

现在你有16秒钟的时间阅读这两题。

6. What did the man think of the meal?A. Delicious.B. Terrible.C. Mediocre.7. Which dish did the woman like best?A. Fish and chips.B. Roast beef.C. Apple pie.听下面一段独白，回答第8和第9两个小题。

Project Management Plan Template-项目管理计划模板

Xavor Corporation<Project Name>Project Management PlanVersion <x.x> [Note: The following template is provided for use in Xavor projects. Text enclosed in square brackets and displayed in blue italics (style=InfoBlue) is included to provide guidance to the author and should be deleted before publishing the document.]Organization Level Template CMM – Xavor Corporation Document #:XVR.0002/PMPVer.:Title:1. Project Management PlanPage #:2 of 17 Revision History[For every revision of this document, provide the revision history that should include the date of revision, version number, description of the changes in the document, and author of the document for that particular version.]Date Version Description Author<dd/mm/yyyy> <x.x> <Details> <Name>Distribution List[State the persons/teams/groups to whom this document should be distributed whenever the document isrevised. Also state the name of their parent organization.]Document Recipient Organization<Role Name> <Organization Name>CMM – Xavor CorporationTitle:1. Project Management PlanPage #:3 of 17 Table of Contents1.Introduction 51.1Purpose 51.2Scope 51.3Definitions, Acronyms and Abbreviations 51.4References 51.5Overview 62.Project Overview 72.1Project Name, Code and Leader 72.2Project Purpose, Scope and Objectives 72.3Assumptions and Constraints 72.3.1Critical Assumptions and Constraints 72.3.2Non-Critical Assumptions and Constraints 72.4Project Milestones 72.5Project Deliverables 72.6Tailoring Guidelines 82.7Software Development Life Cycle 83.Project Organization 93.1Organizational Structure 93.2External Interfaces 113.3Roles and Responsibilities 113.3.1<Organizational Unit Name> 114.Management Process 124.1Work Breakdown Structure (WBS) 124.2Project Estimates 124.2.1Estimation Technique 124.2.2Size 124.2.3Effort 124.3Project Schedule 124.3.1Pre-Development Schedule 124.3.2Development Schedule 124.4Project Phases, Iterations and Releases 124.4.1Project Phases 124.4.2Project Iterations 124.4.3Releases 124.5Project Resourcing 134.5.1Staffing 134.5.2Resource Acquisition 134.5.3Training 134.6Project Budget 13CMM – Xavor CorporationTitle:1. Project Management Plan Page #: 4 of 174.7Project Monitoring and Control 134.7.1Schedule Control 134.7.2Budget Control 134.7.3Measurements 134.8Risk Management Plan 134.9Project Closure 135.Technical Process Plans 145.1User Experience Design 145.2Requirements Management 145.3Analysis and Design 145.4Development Plan 145.5Peer Review Plan 145.6Project Maintenance 145.7Test Plan 145.8Tools, Techniques and Standards 145.8.1Tools 145.8.2Techniques and Standards 155.9Infrastructure 165.10Facilities 165.11Security Plan 166.Supporting Process Plans 166.1Configuration Management Plan 166.2Documentation 166.3Software Quality Assurance Plan 166.4Intergroup Coordination 166.5Communication 166.6Problem Resolution 166.7Subcontractor Management 167.Additional plans 178.Appendices 17CMM – Xavor CorporationTitle:1. Project Management Plan Page #: 5 of 17Project Management Plan1. Introduction[The introduction of the Project Management Plan should provide an overview of the entire document. It should include the purpose, scope, definitions, acronyms, abbreviations, references and overview of this Project Management Plan.]1.1 Purpose[Specify the purpose of this Project Management Plan.]1.2 Scope[A brief description of the scope of this Project Management Plan; what Project(s) it is associated with,and anything else that is affected or influenced by this document.]1.3 Definitions, Acronyms and Abbreviations[This subsection should provide the definitions of all terms, acronyms, and abbreviations required tointerpret properly the Project Management Plan. This information may be provided by reference to the project Glossary.]1.4 References[This subsection should provide a complete list of all documents referenced elsewhere in the ProjectManagement Plan. Each document should be identified by title, report number (if applicable), date, and publishing organization. Specify the sources from which the references can be obtained. This information may be provided by reference to an appendix or to another document. For the Project Management Plan, the list of referenced artifacts may include:•Risk Management Plan•User Interfaces Guidelines•Configuration Management Plan•Software Quality Assurance Plan, etc.]Document Title Publishing Organization<Title> <Organization Name>CMM – Xavor CorporationTitle:1. Project Management Plan Page #: 6 of 171.5 Overview[This subsection should describe what the rest of the Project Management Plan contains and explain how the document is organized.]CMM – Xavor CorporationTitle:1. Project Management Plan Page #: 7 of 172. Project Overview2.1 Project Name, Code and Leader[Specify the project name, project code and project leader (project manager).]Project Name: <Project Name>Project Code: <xxx-xxx>Project Leader: <Name>2.2 Project Purpose, Scope and Objectives[A brief description of the purpose and objectives of this project, and a brief description of whatdeliverables the project is expected to deliver.]2.3 Assumptions and Constraints[A list of assumptions that this plan is based on, and any constraints (e.g. budget, staff, equipment,schedule, etc.) that apply to the project. Make a distinction between critical and non-critical factors.]2.3.1 Critical Assumptions and Constraints[State the critical assumptions and constraints affecting the project.]2.3.2 Non-Critical Assumptions and Constraints[State the non-critical assumptions and constraints affecting the project.]2.4 Project Milestones[Tabular list of major milestones to be achieved during the project, with target dates.]Milestone Target Achievement Date <Milestone Name> <dd/mm/yyyy>2.5 Project Deliverables[Tabular list of the artifacts to be created during the project, with target delivery dates.]Deliverables Target Delivery Date<Deliverable Name> <dd/mm/yyyy>CMM – Xavor CorporationTitle:1. Project Management Plan Page #: 8 of 172.6 Tailoring Guidelines[Specify the tailoring guidelines for the project.]2.7 Software Development Life Cycle[Specify the Software Development Life Cycle that is to be followed in the project.]CMM – Xavor CorporationTitle:1. Project Management Plan Page #: 9 of 173. Project Organization3.1 Organizational Structure[Describe the organizational structure of the project team, including management and other reviewauthorities. This should include identification of all project organizational units and a description of their function and responsibility. A diagram of the organizational structure should also be attached for further illustration.Examples of project organizational units are:•Project Implementation Committee•Project Steering Committee•Project Management Team•Architecture Group•User Experience Design Team•Requirements Team•Analysis and Design Team•Implementation Group•Development Team•Database Management Team•Testing Team•Infrastructure Team•Configuration Management Team•Software Quality Assurance Team, etc.]Organizational Unit Description<Organizational Unit Name> <Description>CMM – Xavor CorporationTitle :1.Project Management Plan Page #:10 of 17Project Team <Project Name>Version: <x.x>Date: <dd/mm/yyyy>Project ManagerProject ManagementTeamProject ArchitectureGroupProjectImplementationGroupUser ExperienceTeamRequirementsTeamSystem Analysis & Design Team System ArchitectureTeam Software DevelopmentTeamTesting TeamDatabase ManagementTeamInfrastructureTeamSoftware ConfigurationTeam Software Quality Assurance TeamCMM – Xavor CorporationTitle:1. Project Management Plan Page #:11 of 173.2 External Interfaces[Describe how the project interfaces with external groups. For each external group, identify the internal/external contact names.]External Organization External Role External RoleHolderResponsibility ofExternal Role HolderInternal ContactRole andPerson<Organization Name> <Role Name> <Role HolderName><Responsibility Details> <Role Name>,<Person Name>3.3 Roles and Responsibilities[Specify the roles, responsibilities and role holders within each organizational unit of the project.]3.3.1 <Organizational Unit Name>Role Responsibility Role Holder<Role Name> <Responsibility Detail> <Role Holder Name>CMM – Xavor CorporationTitle:1. Project Management Plan Page #:12 of 174. Management Process4.1 Work Breakdown Structure (WBS)[List the activities necessary for completing the project.]4.2 Project Estimates4.2.1 Estimation Technique[Specify the estimation method and the reason for its choice. Provide the estimated cost as well as thebasis for those estimates, and the points/circumstances in the project when re-estimation will occur.]4.2.2 Size[State the size of each activity as calculated according to the estimation technique. Units of size may be in LOC, FP, etc.]4.2.3 Effort[Specify the amount of effort required to perform each activity on the basis of the size estimation. Unitsmay be man-hours, man-days, etc.]4.3 Project Schedule[Diagrams/tables showing target dates for completion of iterations and phases, release points, demos, and other milestones. Critical path must be specified. Usually enclosed by reference to MS Project file.]4.3.1 Pre-Development Schedule[This schedule will cater for project planning, requirements, analysis and design activities.]4.3.2 Development Schedule[This schedule will cater coding, testing and deployment activities.]4.4 Project Phases, Iterations and Releases4.4.1 Project Phases[Identify phases and major milestones with their achievement criteria.]4.4.2 Project Iterations[Specify the number of iterations and list the objectives to be accomplished for each of the iterations.]4.4.3 Releases[Brief descriptions of each software release, whether demo, beta, etc.]CMM – Xavor CorporationTitle:1. Project Management Plan Page #:13 of 174.5 Project Resourcing4.5.1 Staffing[Identify here the numbers and type of staff required (including and special skills or experience), scheduled by project phase or iteration. State what resources are critical.]4.5.2 Resource Acquisition[Describe how you will approach finding and acquiring the staff needed for the project.]4.5.3 Training[List any special training project team members will require, with target dates for when this trainingshould be completed.]4.6 Project Budget[Allocation of costs against the WBS and the project phases.]4.7 Project Monitoring and Control4.7.1 Schedule Control[Describes the approach to be taken to monitor progress against the planned schedule and how to takecorrective action when required.]4.7.2 Budget Control[Describes the approach to be taken to monitor spending against the project budget and how to takecorrective action when required.]4.7.3 Measurements[Describe the types of measurements to be taken, their frequency, and responsible workers/entities for this purpose.]4.8 Risk Management Plan[Enclosed by reference]4.9 Project Closure[Describe the activities for the orderly completion of the project, including staff reassignment, archiving of project materials, post-mortem debriefings and reports etc.]CMM – Xavor CorporationTitle:1. Project Management Plan Page #:14 of 175. Technical Process Plans5.1 User Experience Design[Describe the approach that will be adopted with details of processes, procedures, and guidelines to befollowed.]5.2 Requirements[Describe the approach that will be adopted with details of processes, procedures, and guidelines to befollowed.]5.3 Analysis and Design[Describe the approach that will be adopted with details of processes, procedures, and guidelines to befollowed.]5.4 Development Plan[Enclosed by reference]5.5 Peer Review Plan[Specify the work products to be peer reviewed, type of peer review, their frequency, etc.]5.6 Maintenance[Describe details of any software maintenance for the warranty period of the project.]5.7 Test Plan[Enclosed by reference]5.8 Tools, Techniques and Standards5.8.1 Tools5.8.1.1 Project Management Tools[Specify the project management tools that are to be used in the project and the reasons for their selection.Examples of areas to be covered are project planning, project scheduling, project monitoring, statusreporting, measurements, etc. Examples of these tools are MS Project, etc.]5.8.1.2 Requirements Management Tools[Specify the requirements management tools that are to be used in the project and the reasons for theirselection. Examples of areas to be covered are requirements gathering, requirement issue resolution,requirement change management, measurements, etc. Examples of these tools are Rational Requisite Pro, EINS, etc.]5.8.1.3 System Analysis & Design Tools[Specify the system analysis and design tools that are to be used in the project and the reasons for theirCMM – Xavor CorporationTitle:1. Project Management Plan Page #:15 of 17selection. Examples of tools in this area are Visio, Rational Rose, Power Designer etc.]5.8.1.4 Languages[Specify the languages that are to be used for software development in the project and the reasons for their selection. Examples of languages are HTML, Java, etc.]5.8.1.5 User-Interface Development Tools[Specify the tools that are to be used for UI development in the project and the reasons for their selection.Examples of these tools can be Dreamweaver, Flash, etc.]5.8.1.6 Database Management System Software[Specify the database management system software that is to be used in the project and the reasons fortheir selection. Examples of these tools are Oracle, SQL Server, etc.]5.8.1.7 Third Party Software[Specify any third party software that is to be used in the project and the reasons for their selection.Examples are Inktomi, Infranet, etc.]5.8.1.8 Software Testing Tools[Specify the software testing tools that are to be used in the project and the reasons for their selection.Examples of these tools are WinRunner, LoadRunner, etc.]5.8.1.9 Defect and Change Management Tools[Specify the defect and change management tools that are to be used in the project and the reasons fortheir selection. Examples of these tools are ClearQuest, etc.]5.8.1.10 Configuration Management Tools[Specify the configuration management tools that are to be used in the project and the reasons for theirselection. Examples of these tools are ClearCase, etc.]5.8.1.11 Integrated Development Environment[Specify the operating systems (platforms), web servers, application servers, development servers that are to be used in the project and the reasons for their selection. Examples of these tools are Sun Solaris,iPlanet, JBuilder, WebSphere, etc.]5.8.2 Techniques and Standards[Lists the documented project technical standards etc by reference. Examples may be:User-Interface GuidelinesProgramming GuidelinesTest Guidelines, etc.]CMM – Xavor CorporationTitle:1. Project Management Plan Page #:16 of 175.9 Infrastructure[Specify hardware, network connectivity, bandwidth, etc., required in this project. Make a clear distinction about what factors are critical.]5.10 Facilities[Describe the facilities required for the execution of the project. This will cover physical workspace,buildings, etc.]5.11 Security Plan[List down the security consideration e.g. of security can be operating system, access controls tosite/product, physical security considerations.]6. Supporting Process Plans6.1 Configuration Management Plan[Enclosed by reference]6.2 Documentation[Specify the documents that will be produced in the project, what document templates will be used, and any other information pertaining to documentation.]6.3 Software Quality Assurance Plan[Enclosed by reference]6.4 Intergroup Coordination[Describe how different project groups will communicate with one another; specify dependencies, andcommitments.]6.5 Communication[Specify how various workers/units/entities, both within and outside the project team, will communicatewith each other.]6.6 Problem Resolution[Describe the approach for resolving issues in the project, escalation procedures, etc.]6.7 Subcontractor Management[If subcontractors are involved in the project give details of what kind of contractors are required forvarious tasks, the duration for which they are required and how they will be managed.]Organization Level Template CMM – Xavor Corporation Document #:XVR.0002/PMPVer.:Title:1. Project Management Plan Page #:17 of 177. Additional plans[Additional plans if required by contract or regulations.]8. Appendices[Attach any supplementary information.]。

Nipro 产品数据册：单用途翼型肾脏细胞注射器说明书

H O S P I T A L P R O D U C T WING CATHSTERILE, SINGLE-USE WINGED CATHETER WITH INJECTION PORT FOR IV INFUSIONPRODUCT DATA SHEET2PRODUCT COMPLIANCE• CE-marked, Class IIa Medical Devices, Rule 7, MDD 93/42/EEC, UMDNS 10727• Complies with the following norms, directives, and regulations: ―ISO 594-1: 1986 ―EN 980:2008 ―EN 1041:2008 ―EN 1707:1996 ―ISO 7864:2016―ISO 9626:2016 ―EN ISO 10555-1:2013 ―EN ISO 10993-1/4/5/7/11 ―ISO 10993-10:2010 ―EN ISO 11135-1:2014―EN ISO 11607-1:2009 /AMD 2014 ―EN ISO 13485:2016 ―EN ISO 14971: 2012• Labels contain 12 languagesMANUFACTURING DETAILSLegal manufacturer: Nipro Corporation Country of origin: ThailandSTERILIZATION AND SHELF LIFESterilizationSingle-use only Indicator stickerShelf lifeEtO (Ethylene oxide)Each outer box (or shipping carton) contains a chemical indicator that indicates sterility. This indicator comes in the form of a [blue] sticker that turns[red] when sterilized.5 yearsWING CATHSTERILE, SINGLE-USE WINGED CATHETER WITH INJECTION PORT FOR IV INFUSION• Ultra sharp three-beveled needle to minimize discomfort • Radiopaque• Injection port with universal luer fitting • Flexible wings for easy grip• Color-coded wings and hub to indicate gauge• Siliconization of needle minimizes penetration and gliding force• T ransparent ETFE catheter tube for quick visualization of blood in the flashback chamber to facilitate insertion • Flashback in hub means the needle is correctly in the vein • Flashback in catheter means the catheter is correctly in the vein • Latex-free, DEHP-free, PVC-free • Available in 16-18-20-22-24 G• IV infusion duration: hours to several days • For use by healthcare professionals onlyClass IIa Medical DevicesRule 7MDD 93/42/EEC UMDNS 1072701233MATERIALS USEDDEHP-free Latex-free PVC-free PRODUCT RANGE OVERVIEWPACKAGING DETAILSLanguages12 languages on inner and outer box:English (EN), French (FR), Dutch (NL), German (DE), Spanish (ES), Italian (IT), Portuguese (PT), Greek (EL), Swedish (SV), Danish (DA), Norwegian (NO), and Finnish (FI)GlueMaterial: Acrylic resinNeedle (cannula)Material:Stainless steel SUS-304Needle hub Material:Polycarbonate (PC)Catheter hubMaterial: Polypropylene (PP)Port capMaterial: Polyethylene (PE)Catheter tubeMaterial: Ethyltetrafluoroethylene (ETFE)Cutting angleThree-beveledLuer capMaterial: Polypropylene (PP)Cap connectorMaterial: Polypropylene (PP)Caulking pinMaterial: Stainless steel SUS-304One-way valveMaterial: Silicone tubeLubricantMaterial: Silicone PolydimethylsiloxaneMembrane filterMaterial: Random micro glass filterVent fittingMaterial: Polypropylene (PP)Catheter wingMaterial: Polypropylene (PP)Nipro Medical Europe : European Headquarters, Blokhuisstraat 42, 2800 Mechelen, Belgium T: +32 (0)15 263 500 | F: +32 (0)15 263 510 |***********************| D a t a s h e e t - W i n g _C a t h - T H A - 27.N o v 2019Label detailsTransport conditionsClosed and dryStorage conditions Open the packaging only immediately before use to guarantee sterility.。

国开(中央电大)本科《商务英语3》形考任务(单元自测1至8)试题及答案-国开商务英语试题

国开(中央电大)本科《商务英语3》形考任务(单元自测1至8)试题及答案:国开商务英语试题国开(中央电大)本科《商务英语3》形考任务(单元自测1至8)试题及答案 (电大网上形考必备资料) 说明：课程代码：04011。

适用专业及层次：会计学、金融学、市场营销和物流管理本科。

平台考试网址：http：//。

单元自测1 试题及答案题目顺序是随机的，使用查找功能（Ctrl＋F）进行搜索 [题目]— Why did you choose our company？— __________________________ [答案] Sorry, I didn＇t catch that. Would you say it again? [题目]— Can you show me the schedule？—__________________________. [答案] Sure, a tight schedule as usual [题目]— But what do you want me to do there？— ______________________________ [答案] You have to give a presentation on what our products are. [题目]— Morning, Maggie. Could you talk briefly about your career development？— ___________________________ [答案] Thanks for giving me this opportunity. I started as a clerk in the… [题目]— You need to pay a visit to our important customers from _Chamberlain Automation Systems Company in Washington. —_____________________________ [答案] Sorry, can you repeat the name of the company? [题目]______ a vacation is always a good way to get relaxation and refreshment. [答案] Taking [题目]Before I came to America, I __________as an executive secretary with a trading company for one year in Britain. [答案] had worked [题目]Good manners may be varied ________cultural backgrounds. [答案] due to [题目]I led and _________a sales team of 20 people. [答案] motivated [题目]I was __________ devising sales strategies and targets for the West Region. [答案] responsible for [题目]Is ________ possible to take a 10-minute break during the meeting tomorrow morning？[答案] it [题目]Sometimes your body language, gestures and expressions may tell people ______________ about you than the words you use. [答案] more [题目]The following hints may help you stay energized, or at least ___________ until you can make the time for rest. [答案] get you going [题目]When talking to people within your company _________don’t speak your language, you may have to use English. [答案] who [题目]You are the right person for the job that_______ your application ________the advertisement. [答案] relates…to…二、阅读理解阅读下面的文章，根据文章内容给题目选择正确的答案。

碧云天生物技术碘化丙啶产品说明书

碧云天生物技术/Beyotime Biotechnology订货热线：400-168-3301或800-8283301订货e-mail：******************技术咨询：*****************网址：碧云天网站微信公众号Propidium Iodide/碘化丙啶产品编号产品名称包装ST512 Propidium Iodide/碘化丙啶20mg产品简介：Propidium Iodide简称PI，中文名为碘化丙啶。

分子式为C27H34I2N4，分子量为668.40，纯度>95%。

进口分装，常用于细胞凋亡(apoptosis)或细胞坏死(necrosis)的检测，常用于流式细胞仪分析。

包装清单：产品编号产品名称包装ST512 Propidium Iodide/碘化丙啶20mg—说明书1份保存条件：4ºC避光保存。

注意事项：本产品对人体有刺激性，操作时请小心，并注意适当防护以避免直接接触人体或吸入体内。

本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。

为了您的安全和健康，请穿实验服并戴一次性手套操作。

使用本产品的文献：1.Wu G, Liu ZS, Qian Q, Jiang CQ. Effects of Berberine on the Growth ofHepatocellular Carcinoma Cell lines. Medical Journal of Wuhan University. 2008 Jan;29(1):102-105.2.Liu Y, Sheng Z, Liu H, Wen D, He Q, Wang S, Shao W, Jiang RJ, An S,Sun Y, Bendena WG, Wang J, Gilbert LI, Wilson TG, Song Q, Li S.Juvenile hormone counteracts the bHLH-PAS transcription factors MET and GCE to prevent caspase-dependent programmed cell death in Drosophila. Development. 2009 Jun;136(12):2015-25.3.Li DL, Liu JJ, Liu BH, Hu H, Sun L, Miao Y, Xu HF, Yu XJ, Ma X, RenJ, Zang WJ. Acetylcholine inhibits hypoxia-induced tumor necrosis factor-α production via regulation of MAPKsphosphorylation in cardiomyocytes. J Cell Physiol. 2011 Apr;226(4):1052-9.4.Cao X, Deng W, Wei Y, Su W, Yang Y, Wei Y, Yu J, Xu X. Encapsulationof plasmid DNA in calcium phosphate nanoparticles: stem cell uptake and genetransfer efficiency. Int J Nanomedicine. 2011;6:3335-49.5.Meng LY, Liu HR, Shen Y, Yu YQ, Tao X. Cochinchina momordica seedextract induces G2/M arrest and apoptosis in human breast cancerMDA-MB-231 cells by modulating the PI3K/Akt pathway. Asian Pac J Cancer Prev. 2011;12(12):3483-8.6.Zhao Q, Xue Y, Wang JF, Li H, Long TT, Li Z, Wang YM, Dong P, XueCH. In vitro and in vivo anti-tumour activities of echinoside A and ds-echinoside A from Pearsonothuriagraeffei. J Sci Food Agric. 2012 Mar 15;92(4):965-74.7.Tu Z, Ma Y, Tian J, Li H, Akers W, Achilefu S, Gu Y. Estrogen receptor βpotentiates the antiproliferative effect of raloxifene and affects the cellmigration and invasion in HCT-116 colon cancer cells. J Cancer Res Clin Oncol. 2012 Jul;138(7):1091-103.8.Zhou Z, Wan Y, Zhang Y, Wang Z, Jia R, Fan Y, Nie H, Ying S, Huang P,Wang F. Follicular development and expression of nuclear respiratory factor-1 and peroxisome proliferator-activated receptor γ coactivator-1 alpha in ovaries of fetal and neonatal doelings. J Anim Sci. 2012 Nov;90(11):3752-61.9.Jun Fang, Meihu Ma, Yongguo Jin, Ning Qiu, Chan Wang, Guodong Renand Xin Huang. Assessment of Salmonella enteritidis Viability in Egg White during Early Incubation Stages by Fluorescent Staining Method.Asian Journal of Animal and Veterinary Advances. 7: 556-67.10.Zhen-Jun S, Yuan-Yuan Z, Ying-Ying F, Shao-Ju J, Jiao Y, Xiao-Wei Z,Jian C, Yao X, Li-Ming Z.β,β-Dimethylacrylshikonin exerts antitumoractivity via Notch-1 signaling pathway in vitro and invivo. Biochem Pharmacol. 2012 Aug 15;84(4):507-12.11.Tu Z, Li H, Ma Y, Tang B, Tian J, Akers W, Achilefu S, Gu Y. Theenhanced ant iproliferative response to combined treatment of trichostatin A with raloxifene in MCF-7 breast cancer cells and its relevance to estro gen receptor β expression. Mol Cell Biochem. 2012 Jul;366(1-2):111-22.12.Feng C, Xu Z, Li Z, Zhang D, Liu Q, Lu L. Down-regulation of Wnt10aby RNA interference inhibits proliferation and promotes apoptosis in mouse embryonic palatal mesenchymal cells throu gh Wnt/β-catenin signaling pathway. J Physiol Biochem. 2013 Dec;69(4):855-63.13.Hou Y, Chu M, Du FF, Lei JY, Chen Y, Zhu RY, Gong XH, Ma X, Jin J.Recombinant disintegrin domain of ADAM15 inhibits the proliferation and migration of Bel-7402 cells. Biochem Biophys Res Commun. 2013 Jun 14;435(4):640-5.14.Li Q, Zhou X, Shi Y, Li J, Zheng L, Cui L, Zhang J, Wang L, Han Z, HanY, Fan D. In vivo tracking and comparison of the therapeutic effects of MSCs and HSCs for liver injury. PLoS One. 2013 Apr 30;8(4):e62363. 15.Zhou R, Huang W, Yao Y, Wang Y, Li Z, Shao B, Zhong J, Tang M,Liang S, Zhao X, Tong A, Yang J.CA II, a potential biomarker by proteomic analysis, exerts significant inhibitory effect on the growth of colorectal cancer cells. Int J Oncol. 2013 Aug;43(2):611-21.16.Wang Y, Jiang XL, Peng SW, Guo XY, Shang GG, Chen JC, Wu Q, ChenGQ. Induced apoptosis of osteoblasts proliferating on polyhydroxyal kanoates. Biomaterials. 2013 May;34(15):3737-46.17.Yang F, Huang W, Li Y, Liu S, Jin M, Wang Y, Jia L, Gao Z. Anti-tumoreffects in mice induced by survivin-targeted siRNA delivered through polysaccharide nanoparticles. Biomaterials. 2013 Jul;34(22):5689-99.18.Zhou S, Wu H, Zeng C, Xiong X, Tang S, Tang Z, Sun X. ApolipoproteinE protects astrocytes from hypoxia and glutamate-induced apoptosis.FEBS Lett. 2013 Jan 16;587(2):254-8.19.Li R, Luo X, Li L, Peng Q, Yang Y, Zhao L, Ma M, Hou Z. TheProtective Effects of Melatonin Against Oxidative Stress and Inflammation Induced by AcuteCadmium Exposure in Mice Testis. Biol Trace Elem Res. 2016 Mar;170(1):152-64.Version 2016.12.08。

不同酶消化法提取猪原代肝细胞的效果比较

532024.4·试验研究0 引言猪圆环病毒（PCV ）是Circoviridae 科Circovirus 属的一种无囊膜的单链环状DNA 病毒。

在已知的4个血清型中，PCV2为猪易感的致病性病毒[1]。

PCV2感染会诱导宿主免疫抑制引起猪圆环病毒病（PCVD ），包括断奶仔猪多系统衰竭综合征、新生仔猪先天性脑震颤、皮炎与肾病综合征、猪呼吸道病综合征、母猪繁殖障碍等，给全世界养猪业带来较大的经济损失，是世界各国的兽医与养猪业者公认的造成重大影响的猪传染病[2]。

PCV2的感染在猪生长发育的不同阶段有不同的组织嗜性。

但无论是胎儿阶段还是出生后，肝细胞都是PCV2感染和复制的靶细胞。

因此，PCV2也被视为一种能够诱导猪肝炎的病毒[3]。

且PCV2诱导的肝细胞凋亡在PCV2引发的相关病变和疾病的发病机制中具有关键性作用[4]。

因此，方便、快捷地获取大量有活性的猪肝细胞对于研究PCVD 的致病机制具有重大意义。

目前获取肝细胞常用的方法主要包括机械分离细胞法、非酶分离细胞法、离体酶消化法和酶灌流法等[5]。

因此，本试验采用简便、经济、无需特殊设备、仅需部分肝组织的离体酶消化法，比较不同酶消化分离猪原代肝细胞的效果，为一般实验室提取分离大量有活性的猪肝细胞提供参考。

1 材料与方法1.1 材料1.1.1 主要试剂新鲜猪肝组织，Hank's 平衡盐溶液（HBSS ），磷酸盐缓冲液（无菌PBS ），4%多聚甲醛（PFA ），收稿日期：2024-01-27基金项目：国家自然科学基金项目：复杂器官与组织在脾脏内的功能性再生（32230056）作者简介：周徐倩（1999-），女，汉族，浙江温州人，硕士在读，研究方向：组织工程与再生医学。

*通信作者简介：董磊（1978-），男，汉族，安徽阜阳人，博士，教授，研究方向：组织工程与再生医学、生物材料。

周徐倩，董磊．不同酶消化法提取猪原代肝细胞的效果比较[J]．现代畜牧科技，2024，107（4）：53-55．　doi ：10.19369/ki.2095-9737.2024.04.014．　ZHOU Xuqian ，DONG Lei ．Comparison of the Eﬀect of Diﬀerent Enzyme Digestion Methods on Extraction of Porcine Primary Hepatocytes[J]．Modern Animal Husbandry Science & Technology ，2024，107（4）：53-55．不同酶消化法提取猪原代肝细胞的效果比较周徐倩，董磊*（南京大学，江苏南京 210023）摘要：猪肝细胞是猪圆环病毒的靶细胞，简单快速地提取猪原代肝细胞对于研究猪圆环病毒病的致病机制具有重要意义。

ch11_CriticalChainProjectScheduling关键链项目进度计划

a. Other tasks 其它任务 b. Overestimation penalty 过高的惩处 c. Perfectionism 完美主义
3. Multitasking 多任务交叉 4. Path Merging 路经汇聚
Copyright © 2013 Pearson Education
11-08
Goldratt rule of thumb (50%) 高德拉特法则（50%） Newbold formula 纽博尔德方程
➢ Feeder buffers for non-critical paths
汇入缓冲用于非关键路径
Copyright © 2013 Pearson Education
11-12
11-02
Chapter 11 Learning Objectives 学习目标
Distinguish between critical path and critical chain project scheduling techniques.
区分关键路径和关键链项目进度计划技术；
Understand how critical chain methodology resolves project resource conflicts.
“无责备”文化
❖ Subcontractor deliveries & work scheduled ES
将分包商交付工作安排为最早开始时间
❖ Non critical activities scheduled LS
非关键活动安排为最晚开始时间
❖ Factor the effects of resource contention
Managers should

公司加装的电梯特种设备验收流程

公司加装的电梯特种设备验收流程1.电梯加装完成后，需立即通知相关部门进行验收。

After the installation of the elevator, relevant departments should be notified for inspection promptly.2.验收前需要准备好相关资料和文件。

Prepare the relevant documents and files before inspection.3.验收人员应当具备相关技术资格和专业知识。

Inspectors should have the relevant technical qualifications and expertise.4.验收人员应当提前预约验收时间。

Inspectors should make an appointment for the inspection in advance.5.准备好相关图纸、设计方案和施工合同。

Prepare the relevant drawings, design schemes, and construction contracts.6.跟踪电梯加装施工过程，确保符合相关规范和标准。

Track the process of elevator installation to ensure compliance with relevant regulations and standards.7.检查电梯加装工程是否符合国家特种设备安全管理规定。

Check if the elevator installation project complies with the national regulations for the safety management of special equipment.8.清查电梯加装工程过程中的施工记录和验收记录。

Review the construction records and inspection records during the process of elevator installation.9.检查电梯加装是否在规定的时间内完成。

火电项目开工手续办理流程

火电项目开工手续办理流程1.火电项目开工手续办理流程包括哪些步骤？What are the steps of the procedure for handling the start-up procedures of the thermal power project?2.需要先进行土地手续办理。

It is necessary to handle the land procedures first.3.提交土地使用证明文件和申请表。

Submit land use certificates and application forms.4.等待相关部门审批。

Wait for approval from the relevant departments.5.完成土地手续后，进行环保手续办理。

After completing the land procedures, proceed with the environmental protection procedures.6.进行环境影响评估申请。

Submit an application for environmental impact assessment.7.等待环保部门的审批。

Wait for approval from the environmental protection department.8.同时进行用电手续办理。

Handle the electricity use procedures at the same time.9.提交用电申请和相关电力设计文件。

Submit electricity use applications and related power design documents.10.等待电力部门的审批。

Wait for approval from the power department.11.完成环保和用电手续后，进行安全生产手续办理。

湖南省风电风电项目建设主要流程

湖南省风电风电项目建设主要流程1.风电项目规划阶段，需要进行可行性研究和选址调查。

During the planning phase of the wind power project, feasibility studies and site surveys need to be conducted.2.确定选址后，需要进行环境影响评价和项目核准。

After determining the site, environmental impact assessments and project approvals need to be carried out.3.完成项目核准后，立项建议书需要提交相关部门审批。

After the project approval is completed, the project proposal needs to be submitted to relevant departments for approval.4.同时，必须开展土地使用、用地预审和用地批准手续。

At the same time, land use, pre-approval, and land approval procedures must be carried out.5.获取土地使用权后，需进行项目总包、设备采购等招标工作。

After obtaining the land use rights, bidding work for project general contracting, equipment procurement, etc. needs to be carried out.6.招标工作完成后，需要签署合同并组织项目实施。

After the bidding work is completed, contracts need to be signed and the project needs to be implemented.7.项目实施阶段需要进行工程建设和设备安装。

英语作文 The Project

英语作文 The ProjectThe Project。

Introduction:The Project is a crucial part of our academic curriculum, designed to enhance our knowledge, skills, and understanding of a particular subject or topic. It requires thorough research, critical thinking, and creativity to produce a high-quality output. The Project serves as a platform for us to apply what we have learned in the classroom to real-life situations, fostering a deeper understanding of the subject matter.Purpose:The main purpose of The Project is to provide students with an opportunity to delve deeper into a specific area of study and develop a comprehensive understanding of the topic. It also aims to improve our research and analyticalskills, as well as our ability to work independently and manage our time effectively. Additionally, The Project encourages us to think critically, solve problems, and communicate our findings effectively.Process:The process of completing The Project involves several key steps, including selecting a topic, conducting research, analyzing data, and presenting our findings. We begin by choosing a topic that interests us and aligns with our academic goals. Once the topic is selected, we conduct thorough research using a variety of sources, such as books, journals, and online databases. We then analyze the information gathered and draw conclusions based on our findings. Finally, we present our work through a written report, a presentation, or a creative project.Challenges:Completing The Project can be challenging, as itrequires a significant amount of time and effort.Researching and analyzing data can be time-consuming, and presenting our findings in a clear and concise manner can be difficult. Additionally, managing our time effectively and staying organized throughout the process can be a challenge. However, overcoming these challenges allows us to develop important skills that will benefit us in our academic and professional careers.Benefits:Despite the challenges, The Project offers numerous benefits. It allows us to develop a deeper understanding of a specific topic, as well as improve our research and analytical skills. It also provides an opportunity for us to showcase our creativity and critical thinking abilities. Additionally, completing The Project fosters a sense of accomplishment and pride in our work, boosting our confidence and motivation.Conclusion:In conclusion, The Project is an important component ofour academic curriculum that offers numerous benefits. It provides us with an opportunity to delve deeper into a specific area of study, develop important skills, and showcase our knowledge and creativity. While completing The Project may be challenging, the rewards far outweigh the difficulties, making it a valuable learning experience for all students.。

Galactosyltransferase

专利名称：Galactosyltransferase发明人：Strasser, Richard,Steinkellner, Herta,Mach, Lukas,Glössl, Josef,Altmann,Friedrich,Bondili, Jayakumar Singh申请号：EP06450140.6申请日：20060929公开号：EP1905826A1公开日：20080402专利内容由知识产权出版社提供摘要：The present invention provides a method of expressing a plant Lewis-type β1,3-galactosyltransferase in an organism comprising the step of providing the organism with a nucleic acid molecule comprising a sequence A which is defined as being selected from a) a sequence according to SEQ ID NO: 1 with an open reading frame from base pair 1-1932, j) a sequence which is at least 50% identical with SEQ ID NO: 1, k) a sequence which hybridizes with SEQ ID NO: 1 under stringent conditions, or l) a sequence which has degenerated to SEQ ID NO: 1 due to the genetic code, wherein the sequences a) to d) encode a plant protein having Lewis-type β1,3-galactosyltransferase activity, or m) a sequence which is complementary to one of the sequences a) to d), and expressing a protein encoded by sequence A or if the organism comprises a sequence A in its wildtype form overexpressing a protein encoded by sequence A.申请人：Universität für Bodenkultur Wien地址：Gregor Mendel Strasse 33 1180 Wien AT国籍：AT代理机构：Sonn & Partner Patentanwälte 更多信息请下载全文后查看。

初三英语同义词连线超级高级单选题20题

初三英语同义词连线超级高级单选题20题1. She managed to finish the project on time. / He succeeded in completing the project on time.A. startedB. continuedC. finishedD. stopped答案：C。

“managed to finish”和“succeeded in completing”都表达了成功完成某事的意思，其中“finish”和“complete”是同义词。

A 选项“started”是开始；B 选项“continued”是继续；D 选项“stopped”是停止，都不符合题意。

2. The company plans to expand its business. / The firm intends to enlarge its business.A. reduceB. changeC. expandD. keep答案：C。

“plans to expand”和“intends to enlarge”都表示计划扩大业务，“expand”和“enlarge”是同义词。

A 选项“reduce”是减少；B 选项“change”是改变；D 选项“keep”是保持，都不符合。

3. She hurried to catch the bus. / She rushed to catch the bus.A. walkedB. ranC. stoodD. sat答案：B。

“hurried to”和“rushed to”都表示匆忙去做某事，“hurry”和“rush”意思相近，都有赶紧、匆忙的意思。

A 选项“walked”是走路，速度较慢；C 选项“stood”是站立；D 选项“sat”是坐下，都不符合题意。

4. He started to learn English last year. / He began to learn English last year.A. endedB. continuedC. startedD. stopped答案：C。

四川省高标准农田建设项目申报流程

四川省高标准农田建设项目申报流程1.项目立项前，需对土地资源、水资源等进行前期调研和评估。

Before the project approval, it is necessary to conduct preliminary research and assessment on land resources, water resources, etc.2.编制项目申报书，包括项目概况、可行性分析、技术方案等内容。

Prepare the project application, including project overview, feasibility analysis, technical solutions, etc.3.申报书需经过相关部门审批后方可提交。

The application must be approved by relevant departments before submission.4.确定项目建设地点，开展土地使用权手续申请。

Determine the project construction location and apply for land use rights procedures.5.获取土地证和相关手续，并提交相关部门备案。

Obtain land certificates and related procedures, and submit them to the relevant departments for record.6.开展环境影响评价，确定项目对环境的影响情况。

Conduct environmental impact assessment to determine the project's impact on the environment.7.需开展水资源调查评估，明确项目对水资源的利用情况。

Water resources investigation and evaluation are required to clarify the project's use of water resources.8.完善项目的农田规划设计，确保符合高标准农田建设要求。

credit sb. with doing sth.造句 -回复

credit sb. with doing sth.造句-回复1. I want to credit Maria with completing the project ahead of schedule.2. Let's credit John with finding a solution to the complex math problem.3. The team deserves to be credited with achieving record sales this quarter.4. We should credit Sarah with organizing a successful fundraising event for the charity.5. The teacher credited Paul with improving his grades significantly.6. The company should be credited with introducing innovative technology into the market.7. I credit my parents with instilling in me a strong work ethic.8. We should credit the doctor with saving the patient's life througha complex surgery.9. The architect should be credited with designing the stunning new building in the city.10. The team was credited with winning the championship through their exceptional teamwork.11. Let's credit the chef with creating a delicious and unique dish.12. The pioneers are credited with settling and developing the untamed wilderness.13. The scientist should be credited with discovering a groundbreaking cure for cancer.14. We must credit the author with writing a thought-provoking and engaging novel.15. I credit my friend with introducing me to my favorite band, enhancing my musical taste.。

不定式to-do-专项习题

To do 练习题1._______ the project in time, the staff were working at weekends.A. CompletingB. Having completedC. To have completedD. To complete2. Nowadays people sometimes separate their waste to make it easier for it ______A. reusingB. reusedC. reusesD. to be reused3. The play ________ next month aims mainly to reflect the local culture.A. producedB. being producedC. to be producedD. having been produced4. He told us whether _______ a picnic was still under discussion.A. to haveB. havingC. haveD. had5. David threatened _________ his neighbor to the police if the damages were not paid.A. to be reportedB. reportingC. to reportD. having reported6. ------The last one _________ pays the meal.------ Agreed!A. arrivedB. arrivesC. to arriveD. arriving7. ________, you need to give all you have and try your best.A. Being a winnerB. To be a winnerC. Be a winnerD. Having been a winner8. We are invited to a party ________ in our club next Friday.A. to be heldB. heldC. being heldD. holding9. ------Did the book give the information you needed?------Yes. But ________ it, I had to read the entire book.A. to findB. findC. to be findingD. finding10. I feel greatly honored ________ into their society.A. to welcomeB. welcomingC. to be welcomedD. welcomed11. If there is a lot of work ________, I am happy to just keep on until it is finished.A. to doB. to be doingC. doneD. doing12. When I went to the Smiths`, they happened ________ dinner.A. to haveB. to be havingC. to having been havingD. having13. When asked why he went there, he said he was sent there _______ for a space flight.A. trainingB. being trainedC. to have trainedD. to be trained14. ________ the project as planned, we will have to work two more hours a day.A. CompletingB. CompleteC. CompletedD. To complete15. _________ into the college, he had to work hard at all the subjects, some of which he didn’t like.A. In order to admitB. In order to be admittedC. So as to admitD. So as to be admitted16. If the work ________ by the end of the month is delayed, the construction company will be fined.A. being completedB. completedC. to be completedD. is completed17. Sandy could do nothing but _______ to his teacher that he was wrong.A. admitB. admittedC. admittingD. to admit18. ------Is Bob still performing?------I am afraid not. He is said _________ the stage already as he has become an official.A. to have leftB. to leaveC. to have been leftD. to be left19. All these gifts must be mailed immediately _______ in time for Christmas.A. in order to have receivedB. in order to receiveC. so as to be receivedD. so as to be receiving20. He arrived at the station, only _______ the train had left.A. findingB. findC. to findD. found1 / 1。

施工方案英文

Construction PlanIntroductionThe construction plan provides a detled overview of the proposed construction project. It outlines the necessary steps, materials, and resources required to successfully execute the project. This document serves as a guide for the construction team to follow throughout the construction process.ObjectiveThe primary objective of this construction project is to [insert objective here]. The plan ms to achieve this objective by [insert approach or strategy here]. This construction plan will ensure that all activities are carried out efficiently, within the allocated budget, and in compliance with safety regulations.ScopeThe scope of this construction plan includes the following:1.Site preparation: Clearing and leveling the construction site, ensuring proper drnage, and removing any obstacles or debris.2.Foundation construction: Excavating and laying the foundation, including footings and slabs.3.Structural framework: Erecting the structural framework, including columns, beams, and trusses.4.Building envelope: Installing walls, roofs, windows, and doors to enclose the structure.5.Mechanical, electrical, and plumbing (MEP) systems: Installing all necessary MEP systems, including HVAC, plumbing, and electrical wiring.6.Interior finishing: Completing the interior finishes, such as flooring, pnting, and installation of fixtures.7.Exterior finishing: Applying exterior finishes, such as landscaping, pnting, and signage.Project PlanPre-construction Phase1.Project initiation: Identifying the project stakeholders, objectives, and constrnts.2.Permits and approvals: Obtning all necessary permits and approvals before commencing construction.3.Site assessment: Assessing the site conditions, including soil testing and surveying the area.4.Design review: Reviewing the construction drawings and specifications.Construction Phase1.Mobilization: Setting up the construction site, including temporary office spaces, storage facilities, and safety equipment.2.Site preparation: Clearing the site and preparing it for construction activities.3.Foundation construction: Excavating the site and pouring the concrete foundation.4.Structural framework: Erecting the structural components, such as columns, beams, and trusses.5.Building envelope: Installing walls, windows, and doors to enclose the structure.6.MEP systems: Installing all necessary MEP systems, including HVAC, plumbing, and electrical wiring.7.Interior finishing: Completing the interior finishes, such as flooring, pnting, and installation of fixtures.8.Exterior finishing: Applying exterior finishes, such as landscaping and pnting.Post-construction Phase1.Testing and commissioning: Testing all systems andensuring they are operating as intended.2.Quality assurance: Conducting inspections and qualitychecks to ensure compliance with specifications.3.Handover: Transferring the completed project to the clientand obtning final acceptance.4.Project closure: Completing all administrative and financialtasks associated with the project.Resources and TimelineThe following resources will be required to successfully execute this construction project:•Project manager•Architects and engineers•Construction workers and laborers•Construction equipment and machinery•Building materials and supplies•Safety equipment and gearThe estimated timeline for this project is as follows:•Pre-construction phase: [insert duration]•Construction phase: [insert duration]•Post-construction phase: [insert duration]Safety MeasuresSafety is of utmost importance throughout the construction process. The following safety measures will be implemented:•All workers will be required to wear appropriate personal protective equipment (PPE) at all times.•Regular safety meetings and trning sessions will be conducted to ensure awareness of safety practices.•Safety inspections will be carried out regularly to identify and address potential hazards.•Emergency response procedures will be established and communicated to all personnel on-site.•Safety barriers and signage will be installed to prevent unauthorized access to hazardous areas.ConclusionThe construction plan provides a comprehensive roadmap for the successful execution of the construction project. By following this plan, the construction team can ensure that the project is completed on time, within budget, and in compliance with safety regulations. Regular monitoring and updates to the plan may be necessary as the project progresses to address any unforeseen challenges or changes in requirements.。

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The Implementation Process
Termination can be orderly or a “hatchet job” Planning for an implementing an orderly shut down yields better results Who leads the shut down project A special termination manager may be used

That structure absorbs the assets of the project
Termination by Starvation
Termination by starvation involves greatly reducing the budget of a project Used when it is politically dangerous to cancel a project

When to Terminate a Project
Projects take on a life of their own It may be easy to terminate a project that is finished But it can be very difficult to terminate a project prior to its completion
1.
2. 3. 4. 5.
Successful Unsuccessful Changes in environment Take too long Murder

When work on a project stops, some organizational work continues
Termination by Addition
Applies to an in-house project When the project is successful, it is institutionalized While the project goes away, project personnel and assets are transferred to the new business
The Final Report—A Project History
1. 2. 3. 4.
5.
Project performance Administrative performance Organizational structure Project and administrative teams Techniques of project management

Non-Technical Reasons for Termination Political Cross-cultural Senescence

The Termination Process
1. 2.
Must first decide to terminate Next, must carry out the decision to terminate
1. 2. 3. 4.
Termination by extinction Termination by addition Termination by integration Termination by starvation
Termination by Extinction

Extinction occurs in any scenario where the project goes away

Things to Do
1. 2. 3. 4. 5. 6. 7. 8. 9.
Insure tasks are completed Notify the client Finish the paperwork Send out final invoices to the client Redistribute resources Clear with legal Determine what records to keep Assign support Close the project books
The Decision Process
Sunk costs are not relevant to the decision about terminating a project The main issue is if the cost to finish is more/less than the value of the project when finished

Termination Integration
The most common way to terminate a project The project comes into the business

–
It is absorbed into the existing structure
Chapter 13
Completing the Project
Copyright 2009 John Wiley & Sons, Inc.
Introduction

All projects end
–
–
The objectives have been completed It no longer makes sense to finish

Some teams move on to other projects Other times, members goes their own way The client may be happy, mad, or anywhere in between
The Variables of Project Termination