市场营销专业英语论文[优秀范文5篇]

第一篇：市场营销专业英语论文

《市场营销专业英语》

（三）Ping An Insurance(Group)

Ping An Insurance(Group)Company of China, Ltd(“Ping An” or “the Company” or “the Group”)was established in 1988 in Shekou, Shenzhen.The Group is the first insurance company in China to have a shareholding structure.Today it has developed into an integrated financial services conglomerate with insurance, banking, and investment businesses at its core.The Group’s stocks are listed on the Hong Kong Stock Exchange under stock code “2318” and on the Shanghai Stock Exchange under”601318“.Ping An's mission is to fulfill its commitments to its investors, customers, employees and society.Ping An's goal and commitment to investors are to produce stable returns and boost asset values.The commitment to customers is to provide the best services with integrity.The commitment to employees centers on being a responsible employer and providing the conditions for better living and career development.Last but not least, Ping An's commitment to Society is based on Ping An's desire to give back to the society and contribute to the development of the country.Ping An focuses on value creation and the pursuit of excellence.The Group advocates that each employee be a person of integrity and value by highlighting individual virtues such as ”honesty, trustworthiness, entrepreneurship, and achievement,“ and team qualities such as ”unity, vitality, learning and innovation“.The mechanisms of ”competition, motivation, elimination“ are implemented, together with a distinctive, professional, advanced and long-term business philosophy.Ping An's vision is to become an internationally leading integrated financial services group by leveraging its insurance, banking and investment businesses to achieve long-term, stable and healthy growth while creating sustainable and greater value for its stakeholders.Through its subsidiaries such as Ping An Life Insurance Company of China, Ltd.(”Ping An Life“);Ping An Property & Casualty Insurance Company of China, Ltd.(”Ping An Property & Casualty“);Ping An Annuity Insurance Company of

China, Ltd.(“Ping An Annuity”);Ping An Health Insurance Company of China, Ltd.(“Ping An Health”);Ping An Bank Co., Ltd.(“Ping An Bank”);Credit Guarantee Insurance Business Unit of Ping An Property & Casualty Insurance Company of China, Ltd.(Ping An Consumer Finance), or PA P&C CGI Business Unit(PACF);China Ping An Trust & Investment Co., Ltd.(“Ping An Trust”);Ping An Securities Co., Ltd.(“Ping An Securities”);Ping An of China Securities(Hong Kong)Co., Ltd;Ping An Asset Management Co., Ltd.(“Ping An Asset Management”);Ping An of China Asset Management(Hong Kong)Co., Ltd.(“Ping An Asset Management, Hong Kong”);and Ping An Futures Co.Ltd.(“Ping An Futures”), the Group provides comprehensive products and services in insurance, banking and investment to more than 51 million retail customers and two million corporate clients.Ping An has about 417,000 life insurance sales agents, over 83,000 full-time employees, and a network of more than 3,800 branch offices.As at 31st December 2009, consolidated total assets and total equity of the Group were RMB 935.7 billion and RMB 91.7 billion, respectively.The Group posted total income of RMB 152.8 billion and net profit of RMB 14.5 billion in 2009.Ping An Life is the second largest life insurer in China by insurance income, while Ping An Property & Casualty is the second largest property and casualty insurer in China.In 2008, Ping An was included in Fortune Magazine’s “Global 500” list, ranking No.1 among China’s non-State owned enterprises.In June 2009, Ping An was included in The Financial Times Global 500 companies list, ranking second among insurers globally, eighth among Chinese enterprises and first among China’s non-State owned firms.In Forbes Magazine’s Global 2000 list of companies, Ping An was ranked No.141.As a professionally-managed and world-class organization, Ping An is the first Chinese insurer to have introduced foreign investors into its shareholding structure.Operating along the principle of “Group Holdings, Segment Operation, Segment Supervision, Group Listing”, Ping An has been able to ensure the Group as a whole is moving towards its goal, upon the foundation that unites strategy, branding and

corporate culture.Its financial platform is the most integrated among China’s financial services sector.Its national back office support operation in Zhangjiang, Shanghai is a leading financial backend support center in Asia.Through this center, Ping An has established a systematic backend operation.By having in place a service infrastructure that includes telephony, the Internet and customer service staff, a full range of financial services can be delivered to customers in a professional, standardized and comprehensive manner.Value-added services are also being delivered to customers through Customer Service Day and other innovative services such as Wanlitong and One Account.According to authoritative surveys in 2009, customer services at Ping An Life and Ping An Property & Casualty were ranked number one in their respective areas by customers.As a founding member of the Chinese Federation of Corporate Social Responsibility, Ping An takes it a mission to serve as a leader in the area.Ping An abides by the law in its operations and declares its tax in an honest and transparent manner.Through offering charitable assistance in areas such as education, Red Cross and disaster relief, Ping An continuously fulfills its moral and social obligations to the society.By the end of 2009, Ping An has participated in the building and planning of 100 Hope Primary Schools in China, of which 84 schools have been built(including 32 that are being built).In 2009, Ping An also donated RMB 10 million to set up “Ping An Hope Scholarship” for Ping An Hope Primary Schools across China.For six consecutive years Ping An has held its education programs giving away study grants worth a total of RMB 7.74 million to 2,285 students.In 2009, Ping An launched “Start a Business” competition and donated RMB 5 million to the China Youth Development Foundation for the establishment of Ping An Foundation.Ping An has donated over RMB 1 million each year over the last six years in support of ”China Children Protection and Education Program“.Ping An has mobilized blood donation among its staff for eight consecutive years with the blood donated reaching 8.36 million cc.Ping An provides accident and critical illness insurance coverage for those who donated hemopoietic stem cells(HSC)starting in 2003 and

until 2010.The premiums of the donated policies are expected to exceed RMB 700 million.For these charitable acts, Ping An has been widely recognized in a number of major awards and titles, including ”China Most Admired Companies“ for eight consecutive years, ”China Best Corporate Citizens“ for five consecutive years, and ”Most Responsible Companies" for three consecutive years.

第二篇：专业英语论文

课程论文

院系 物理与电子工程学院专业 电子信息工程班级 电工1111班学生姓名 简东阳学号2011140131任课教师 罗倩倩

2013 年12月26日

Switching technology present situation and development trend

Jiandongyang

Hubei university of Arts and Sciences

420683a0196.cdb@sina.cn

Abstract：Direction in the future, more extensible features will become the important characteristics of a new generation of Internet, the exchange of technology, will be as a key technology to host this extensible features.Switching technology in scale, function, performance, safety and service and other aspects have better extensibility features, will be a new generation of Internet architecture in the future.Keywords:Network technology;The Internet;Switching technologyThe present situation of the exchange of technologyThe rapid development of network technology, Ethernet is always dominant.However, with the deepening of the network technology application, the network development scale and speed are speeding.Not only the speed of the LAN got improved, gigabit Ethernet also gradually became popular.attention to network of high reliability, high performance, easy maintenance and reduced cost, and these factors is dependent on the using kind of networking technology;Equipment vendors would on the basis of not harm the interests of the user network functions and pursue the considerable profit.About the network, there have been a mature traditional layer 2 switching technology, and the function of the routing layer 3, and a high service level of network layer 7.switching technology of A single traditional layer 2.More traditional layer 2 switching technology, is a special single, it contains only the data link layer forward.Layer 2 switching technology is mostly used for small local area network(LAN), the machine Numbers keep in 20 or 30 sets of the following is the security of the network environment, fast switching function, multiple access port, low price which the three layer 2 switching technology has has solve the problem that the small network users.2.1 The third layer switching technology with routing function

In order to solve the problems of the broadcast domain, for the first time in 1997 the third layer switching technology with routing function appeared.Layer 3 switching technology has now become a main force of the construction of multiple business fusion network.Using a large local area network(LAN), can increase the dangers of broadcast storm, in order to reduce the harm, we must divide a large local area network(LAN)according to the function or the region into multiple small local area network(LAN), so that it can reduce the harm of broadcast storm.If only use layer 2 switching technology, a large number of visits between subnets cannot be achieved.Modified by the layer 3 switching technology(layer 3 switches)is very good to solve the problem.Today, layer 3 switching technology is more and more mature, and also in constant development and innovation, it occupies a more and more important status in the network.2.2 layer 7 switching technology with the function of network service

Unlock every layer encapsulates of each data packet, identify the application layer information at the same time, is the functions of layer 7.Now, many service providers focus gradually shifted to how to make full use of broadband resources and how to manage the Internet application, content.layer 7 switching technology can handle network application layer data forwarding well, the network service level can be improved and it also can make the network administrators and make the users have a lower cost to optimize the network resources.Whether it is business to consumer e-commerce, market sales of unmanned automation, on-line customer support, personnel construction and planning, customer service, firewall load balancing broadband and intelligent filter content and so on, can be dabble in by layer 7 switching technology.The development trend of switching technology

Research in recent years the development of the exchange network architecture and related algorithms, is mainly to study of switching technology.Switching network is roughly by single-stage exchange structure and multiple exchange structure of two kinds of structure.Single-stage exchange in the performance and scale structure is hard to expand and develop, so the new exchange structure instead of a single-stage exchange structure is imperative.Compared with single-stage exchange structure, multi-level exchange structure has a very big advantage is the extensions is good, if you want to achieve more mass exchange network, multistage switching structure will be the first choice, and it is also considered very promising.The system structure of open-ended standardization and data packet based will gradually become the next generation network of the new definition, this is also to develop more and more effective applications and business functions, at the same time can quickly adapt to the market.achieve higher performance and greater scale, is the main goal of the studying exchange technology.In order to achieve this goal,it is bound to reinforce the research of the project, the developing trends in the future may have the following:

(1)Multistage switching structure instead of the single stage exchange structure;

(2)Reduce the internal blocking of multistage switching structure;

(3)Build more mass exchange network.The application of new technology requires a lot of time to continue to improve and perfect.According to the report, the development of next generation network technology is gradually mature and stable.Believe that the exchange technology will be developed in network technology and business development of the evolving process.And in the business operation and competition in the future, differential operators will also play an increasingly important role.Conclusion

High technology development change rapidly, more and more quickly.Switching technology has become more intelligent,order to reduce the cost, and make the network of high reliability, high performance, easy to maintain and extend the guarantee,Switching technology from the initial layer 2 s developed to the layer 3 and now 7 layer,it is still in the continuous development and expansion.Believe that the network intelligent management is not far from us.Although there is a problem but it will not hinder the exchange of technology application and development, operators and equipment suppliers will make these problems get very good settlement,constantly positive test, unceasingly perfect, eventually there will be a good result.References:

[1] Feng Huibin WengYingJing.Based on the application of modern switching technology course teaching reform research [J].Journal of education teaching BBS, 2012(S4).[2] Wu Wenjing.The next generation network of the soft exchange technology [J].Journal of railway signal engineering, 2005(01).

第三篇：专业英语论文翻译

A Parallelization Cost Model for GPU

GPU并行成本模型 2009137127 周幼兰

Abstract-Using GPU for general computing has become an important research direction in high performance computing technology.However, this is not a lossless optimization method.Due to the impact of device initialization cost, data transmission delay, specific characteristics of programs, and other factors, the general computing on GPU may not always achieve the desired speedup, and sometimes results in program execution performance degradation.On the basis of in-depth analysis of GPU internal processing mechanisms, the main factors affecting GPU implementation performance are pointed out, and a parallel cost model for GPU based on static program analysis is proposed to provide judgement basis for using GPU in general computing.摘要：在高性能计算技术领域，使用GPU执行通用计算已成为一个重要的研究方向。但它并不是一种无损优化方法。由于受设备初始化成本、数据传输延迟、程序本身特征和一些其他因素的影响，基于GUP的通用计算不可能总是达到预期的加速，有时还会导致程序执行性能下降。在对GPU内部处理机制进行深度分析的基础上，得到影响GPU执行性能的主要因素，且得出基于静态程序分析的GPU并行成本模型为GPU在通用计算中的使用提供了判断依据的结论。

I.INTRODUCTION Graphics processing Unit(GPU)has developed at a speed much faster than the Moore’s Law in recent years, not only improving image processing, virtual reality, computer simulation, and the development of related applications, but also providing a good running platform for general-purpose computing using GPU beyond graphic processing.The application of GPU in general-purpose computing makes a series of new challenges faced by the development of the high performance computing technology.Currently, in order to reduce the programming complexity of GPU in general-purpose computing, many GPU manufacturers and research institutions proposed a number of programming languages and programming models close to traditional programming methods, but different styles, such as Brook+[1], CUDA[2] and OpenCL[3], etc.However, as GPU has its own specific characteristic of hardware architecture and development, programmers must have a high level of expertise.Using GPU for general-purpose computing, the increased performance achieved in large part depends on the hardware knowledge and programming skills of programmers.At present, the most studies for the GPU’s parallelism both at home and abroad directly rewrite and transplant programs on the basis of original serial programs.Because software programmers often lack a deep understanding of the hardware platform and have no corresponding capability of programming hardware programs, program transplantation lead to the increased effects achieved of all kinds of applications accelerating general-purpose computing using GPU have obvious difference [4-6].Program performance analysis technology as a basic method of understanding program behavior, plays an important role for comparing the performance difference between different program implementation, identifying performance bottlenecks of programs, and understanding the hardware resource utilization, and is the important part of development and optimization of high

performance computing programs [7].How to take advantage of program performance analysis technology and combine the architecture characteristics of CPU and GPU to guide the planning and optimization of parallel programs so that a variety of computing resources of CPU and GPU are fully utilized is a problem worthy of study using GPU in general-purpose computing at present.简介：

近几年来，计算机图形处理器（GPU）比摩尔定律发展得更迅猛，这种发展不仅体现在改善图形处理、虚拟现实、计算机模拟以及相关运用方面，还体现在为使用GPU作图形以外处理的通用计算提供了良好的运行平台。

GPU在通用计算方面的运用面临着高性能计算技术发展的一系列新挑战。目前，为了减少通用计算中GPU编程复杂性，许多GPU制造商和研究机构提出了一系列编程语言和编程模式，这些编程模式类似于传统德编程方法，但是具有不同的编程风格，例如Brook++、CUDA以及OpenCL等等。然而，当GPU拥有自己独特的硬件结构和发展特征时，与此同时程序员必须拥有较高水平的专业技能。通用计算中使用GPU来提高性能很大程度上取决于程序员的硬件知识和编程技术。迄今为止，国内外对于GPU的平行性研究大多数是直接写入和将程序直接植入到原始串行程序基础上。由于软件程序员往往对硬件平台缺乏深层次的了解，并且没有相应的编写硬件程序的能力，所以程序植入导致了各种各样的运用程序的增加效果有着明显的差异，这些运用程序加速了使用GPU的通用计算。作为一种理解程序行为的基本方法，程序性能分析技术在比较不同程序执行的性能差异、找出程序的性能瓶颈和了解硬件资源的利用率方面扮演着重要角色，而且它还是高性能计算程序的重要组成部分。如何利用程序性能分析技术以及如何结合CPU和GPU的体系特征来引导平行程序的规划和优化以至CPU和GPU的大量计算资源得到充分利用，是目前GPU在通用计算方面一个值得探讨的问题。

V.CONCLUSION There are a number of studies on the applications related to using GPU in general-purpose computing.The most researches focus on using GPU to improve the execution performance of applications.However, how to measure the costs of GPU at runtime is less discussed.From the perspective of GPU’s internal operation mechanism, analyze the key factors affecting the GPU implementation performance, and propose a cost test algorithm based on static program analysis.The results obtained by experiments show that the algorithm proposed can estimate relatively accurate GPU execution performance, and thus provide a useful reference for transplantation of traditional high performance computing to GPU.结论：

与使用GPU作通用计算相关运用的研究实例比比皆是，且大多数研究将使用GPU来提高运用程序的执行性能为研究中心，但是至于怎样衡量GPU运行时的成本却很少有人提及。本论文从GPU内部运行机制出发，分析影响GPU执行性能的主要因素，提出基于静态程序分析的成本测试算法。实验所得结果说明所提算法能够相对精确地评估GPU执行性能从而为传统的高性能计算的在GPU中的移植提供了一个实用性的参考。

Comment: Cost model has been widely used in the computer field as a way to evaluate whether a program is excellent or not.And quite a number of researchers in computer field has proposed various kinds of cost models for GPU, of which most are under certain conditions.But this paper give us a new perception of cost model for GPU in general-purpose computing.The parallelization cost model for GPU can be generally applied for many application areas.The authors attempt to estimate the cost comprehensively.Actually they take the GPU initialization cost, transmission of data cost as well as the program execution cost into consideration.Each aspect use a special algorithm to calculate the cost.And the cost is measured by the time of each aspect.In this paper ,we can get through the authors’ idea very well ,because they are well organized in form and shown clearly in graphs, charts as well as in equations.The parallelization cost model is more accurate, flexible and portable than models of the past.So we should learn the method they study a topic.That is to consider a question in a broad perspective.And if we keep thinking in this way ,our ability of doing scientific research will be greatly improved.At the same time, we should keep close watch on the field of GPU.As it is such an important processor that it is used widely used on modern computers.It is even considered the core graphics processor of computers.And it develops faster nowadays as the need rises.

第四篇：专业英语论文翻译

嵌入式系统研究

专业英语期末考查

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嵌入式系统研究

1前言

智能软件Agent是能够为用户执行特定的任务、具有一定程度的智能、能够自主的执行部分任务并以一种合适的方式和环境相互作用的软件程序。Agent 有自主性、响应性、学习能力和社会性等特性。这使得它适合在高度动态的环境下做出及时的响应。

嵌入式技术和新一代移动通讯网络的发展使得嵌入式智能设备大量的涌现。这些设备的大都具有嵌入式操作系统的支持, 并运行着越来越丰富的应用程序。如何对这些应用程序进行测试，就成为一个需要研究的课题。本文将Agent技术引入嵌入式智能设备的测试中，使用目标设备Agent, 测试控制Agent, 网络环境Agent分别模拟和处理测试设备，测试工程师和测试环境的复杂性，利用Agent自身具有的特点，提出了一种有效的自动化测试的方法。

2相关的研究及本文的思路

Agent 所具有的自主性、响应性、学习能力和社会性等特性，使得它很适合处理复杂测试系统中的问题。将Agent技术应用于测试领域已经有一些相关的研究，下面是具体的介绍。

Jeongeun Choi 和 Byoungju Choi [1]提出了一个基于Agent技术的测试工具，它通过使用用户接口Agent来处理和测试者的交互，使用测试用例选择Agent来进行测试用例的选择，并使用回归测试Agent进行回归测试，从而很好的实现了自动化的软件测试。但是他们提出的测试用例选择技术仅能在大量已有的测试用例中选择最佳的用例，不能减少编写测试用例本身的复杂性。

另外一些研究将Agent 技术应用到了某一领域的测试中。Yu Qi、David Hung 和 Eric Wong [3] 提出了一个基于Agent 技术的Web 应用程序测试方法。专业英语期末考查

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他们使用Agent技术降低了网络应用测试的复杂性。他们的方法不仅仅适用于Web应用程序的测试，也适合于嵌入式智能设备的测试。

在实际测试过程中，测试人员很大的一项任务仍然是编写各种测试用例，要真正的提高测试效率，就要提高测试脚本的通用性，减少测试脚本的变化[2] [4]。常见的嵌入式智能设备测试工具(比如TestQuest)使用图像比对来判断目标设备的状态, 这种方法虽然实现了非侵入性的测试,但是存在两个问题,图片的抓取和传送消耗了大量测试资源,不同手机的用户界面风格变化很大,这使得测试脚本在用来进行新设备的测试时需要对脚本进行维护.然而,实际的设备中,当重要的事件发生时,系统会产生敏感事件来激励相关模块进行处理.如果能够在测试过程中捕获这些敏感事件,就能更加高效的进行测试.并且,只要操作系统相同,敏感事件的捕获方式也是一样,测试用例不会因为界面的变化而变化。本文用目标设备Agent抽象和捕获测试过程中的敏感事件，并将敏感事件发送给测试控制Agent。而测试控制Agent收到敏感事件后，根据自己的知识，采取相应的动作的机制（如异常处理，重新调度测试等）加以处理。这正好可以作为Agent的推理规则，用Agent的智能性来屏蔽测试过程的复杂性。此外，考虑到目标设备处于复杂的网络环境中，我们利用网络环境Agent来控制目标设备所处的网络信号，从而实现对设备所处网络环境的控制。基于Agent的测试系统

3.1 总体结构

测试系统的最终目标是实现一个移动数字终端的自动化测试平台，能够对移动数字终端上的系统软件和应用层软件建立模块化、可重用的测试脚本库，提高各种黑盒测试的效率，自动化测试结果的生成，并实现测试结果的回放。

如图1所示：测试平台分为四层，包括用户接口层，测试控制层，通讯层和设备Agent层。其中，过程监控模块，测试环境控制模块和设备Agent层采用Agent的思想进行设计，以更好的支持嵌入式设备的测试。

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用户接口层测试管理与配置脚本编辑器虚拟手机测试控制层测试结果验证测试资源库测试脚步执行测试环境控制测试过程监控Agent通讯层设 备agent层Synblan Agent蓝牙 GPRS 3G AT视窗的移动Agent定制AgentAT 接口

图1 测试系统的整体架构

3.2 系统基本执行流程

测试过程是对真实用户使用手机时“输入－反馈”模型的一个模拟。系统的基本的使用用例如下，首先用户通过脚本编辑器或者虚拟手机创建测试脚本并进行基本的配置，然后过程监控模块调度脚本开始执行，当脚本解释器解释执行脚本语句时，脚本解释器通常使用通讯模块向手机发送模拟按键，然脚本解释器进入等待状态，目标设备Agent从通讯模块得到按键，执行按键动作，截取屏幕并返回给测试控制Agent，或者如果手机有敏感世事件的发生（如来电，短信，异常等）也会反馈给测试控制Agent，测试控制Agent接到数据，再次激励脚本运行，并同时将测试结果输入存入测试资源库。脚本继续运行下面的语句，直到运行结束。3.3 目标设备Agent

目标设备Agent驻留在被测试的智能设备中，实现对目标设备的建模。本系统中它接收从PC机中接收到的控制命令，然后在智能设备中进行相应的操作，包括模拟键盘事件，截取屏幕并返回给PC，以及根据知识库中的配置的测试目标，通过推理，有所选择的将必需的状态信息主动通知测试宿主机。3.3.1设备Agent 依赖的决策规则

系统事件报告决策规则（SERD-RULE）这个决策规则决定设备Agent在获取到状态信息时，是否将状态信息反馈给上层系统。

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异常处理规则（EM-RULE）

这个决策规则决定了当Agent发现测试中出现异常，要采取的处理方式。3.3.2 Agent 属性

目标设备Agent具有下面的属性： ⑴ 自主性

目标设备Agent使得测试用户无需关注设备的细节，它是设备功能和状态的一个很好的抽象，通过目标代理Agent，上层测试框架只需要执行相应的虚拟操作，则目标设备Agent会自动将这些虚拟操作映射为对手机的实际操作，并且手机的响应和其它状态也能够自动的反馈给客户，这使得上层测试和底层的设备具体操作无关。

⑵ 智能性

当目标设备Agent获取到一个变化的被测系统状态或信息时，它会根据推理规则，仅仅把和本次目标相关的信息发送给测试控制Agent。目标设备Agent还能够根据异常处理规则，合理的处理测试中出现的异常。3.4

测试控制 Agent

测试控制Agent 实现了对测试过程的建模。它主要实现三个目标： ⑴ 在测试过程中，对测试人员感兴趣的事件进行捕获，当这些敏感事件发生后，系统立刻执行预定义的动作，并为测试提供准确和详尽的日志。

⑵ 通过和脚本解释器交互，实现异步的事件通知和交互测试用例的执行。⑶ 处理测试过程中的异常，使得测试过程可以自我修复，进而多个脚本能够按照预先的调度正常运行.3.4.1测试控制Agent的决策规则

⑴ 事件－条件－行动规则（ECA－RULE）

这是测试控制Agent所依据的核心规则，它定义了某个测试所关心的事件，并根据比对条件，采取相应的动作。

⑵ 异常处理规则（EM-RULE）

测试控制Agent也遵守异常处理规则，它能够检测测试过程中发生的异常事件，并执行相应的解决方案，使得测试能够顺利的进行。

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3.4.2测试控制Agent 的属性

⑴自主性

自主性使得测试人员无需过分关注测试的细节，测试人员仅仅需要选择测试目标，测试控制Agent就能根据知识库中的信息，自动生成ECA规则表，并在脚本运行时，按照ECA规则，捕捉敏感的事件并执行相应的操作。

⑵智能性

测试控制Agent能够根据用户选择的测试目标，自动生成ECA规则表，并根据ECA规则，进行推理并采取相应的动作。测试控制Agent还能根据异常处理规则来处理测试过程中发身的异常，保障测试过程的自动，平稳进行。3.5 网络环境Agent 网络环境Agent 实现了对网络环境的模拟和控制。为了测试手机的射频性能，通常使用基站模拟系统来提供设备所处的无线网络环境，并且利用编程接口，通过软件对设备的环境进行控制，使得复杂网络环境下对目标设备的测试成为可能。系统评估

为了评价本系统的有效性，我们设计了一个比较实验。试验在信息产业部泰尔实验室真实的测试项目中进行。我们将测试工程师分为两组，第一组使用本文的系统MobileTest进行测试，第二组使用业界著名的TestQuest Pro 进行测试。测试的依据是中国通讯行业标准的数字移动台测试部分。测试的内容是根据这两个系统各自已经建立好的回归测试用例对新的智能手机进行回归测试，从而比较这两个测试工具的测试效率和脚本的可维护性。

实验度量了两个工具在测试不同种类的测试用例时，自动化执行的用例所占的百分比。从表中可见，虽然TestQuest 在功能测试上有更高的自动化率，在压力测试，多状态测试，多任务测试，临界测试和总计中，MobileTest有更好的测试覆盖率。这是因为在后四种测试中，测试用例需要不断追踪系统的状态，使用图片比对机制的TestQuest 很难完成这些工作，而MobileTest 却能使用Agent返回的敏感事件来进行处理。此外，MobileTest 完成测试任务的时间时15天，比TestQuest 的测试效率要高。

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5总结

本文提出了一个基于Agent技术的嵌入式智能设备的测试方法，它用Agent技术对嵌入式智能设备测试过程中的被测试设备，测试者和网络环境进行模拟和控制，很好的屏蔽了嵌入式智能设备的复杂性，提高了测试的效率和脚本的可重用性。将来的研究工作中，我们会进一步拓展整个测试系统，使之能够支持整个测试的生命周期。

本文作者的创新点：本文将软件Agent技术引入到嵌入式智能设备的自动化测试中，利用Agent的特性，很好的屏蔽了嵌入式智能设备测试中，测试者，待测设备，网络环境的复杂性，提高了自动化测试的效率。

来源于：《 Journal of Electronic Science and Technology》

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附：英文原文

Embedded system research 1 Introduction

Intelligent Software Agent is the ability to perform specific tasks for the user, with a certain degree of intelligence, able to perform some tasks and autonomy in a proper manner and environment interactions software programs.Agent has autonomy, responsiveness, learning and social and other characteristics.This makes it suitable for highly dynamic environment to make a timely response.Embedded technology and the development of new generation mobile communication networks makes the emergence of a large number of embedded intelligent devices.Most of these devices with embedded operating system support, and run an increasingly rich applications.How to test for these applications to be a need to study.This article Intelligent Agent technology into embedded devices test, using the target device Agent, test control Agent, Network Agent and treatment were simulated test equipment, test engineers and test complexity of the environment, the use of Agent has its own characteristics, proposed An effective automated testing.2 Related research and ideas of this article

Agent possess autonomy, responsiveness, learning and social and other features, makes it very suitable for handling complex problems in the test system.Agent technology in the test area will already have some relevant research, the following is a specific description.Jeongeun Choi and Byoungju Choi [1] proposed a testing tool based on Agent technology, which through the use of Agent to handle user interface and test those interactions, the use of Agent to carry out the test case test case selection choices, and use the Agent for regression testing regression testing, so a good automated software testing.But they can only be made in a large number of test selection techniques select test cases has been the best use cases, write test cases can not reduce its complexity.专业英语期末考查

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Other studies will Agent technology applied to a field test.Yu Qi, David Hung and Eric Wong [3] proposed a Web-based Agent technology application testing methods.Agent technology to reduce their use of the network application testing complexity.Their method not only for Web application testing, but also for the testing of embedded intelligent devices.In the actual testing process, testers great task is still the preparation of various test cases, to really improve test efficiency, we must improve the general test scripts, reduce test script changes [2] [4].Common test tools for embedded smart devices(such as TestQuest)using image comparison to determine the status of the target device, although this method to achieve a non-invasive test, but there are two problems, the image capture and transmission consumes a lot of test resources, the different style of phone's user interface has changed dramatically, which makes the test script used for testing new equipment required for maintenance of the script.However, the actual device, when an important event occurs, the system will generate sensitive events to encourage the relevant module for processing.If during the test capture these sensitive matters, will be able to test more efficient.And, as long as the operating system the same sensitive way to capture the same event, the interface test cases will not change.In this paper, the abstract and the Agent target device during the test capture of sensitive events and control of sensitive events are sent to test Agent.Agent receives the test control sensitive incident, according to their knowledge, take appropriate action mechanisms(such as exception handling, re-scheduling the test, etc.)to be addressed.This is precisely the inference rules as Agent, with the intelligence of Agent to shield the complexity of the testing process.In addition, given the target device is a complex network environment, we use the Internet Agent to control the target device environment in which the network signal, enabling the network environment in which the control device.3 Agent-Based Test System 3.1 The overall structure

Test system is to achieve the ultimate goal of a mobile digital terminal's automated testing platforms to the mobile digital terminal on the system software and 专业英语期末考查

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application layer software build modular, reusable test script library, to improve the efficiency of various black box testing, automation the generation of test results and test results to achieve playback.Figure 1: Test platform divided into four layers, including user interface layer, the test control layer, communication layer and the equipment Agent layer.Among them, the process monitoring module, test equipment, environmental control module and the use of Agent Agent layer design ideas to better support embedded devices testing.The user interface layerTesting management and configurationScript editorVirtual cell phone

TestControl layer Test results show Testing steps executiontestrepository Test environment control Testing process monitoring AgentCommunications layer Set for Agent layerBluetooth GPRS 3G ATSynblan Agent Windows mobile AgentCustomization AgentAT 接口Figure 1 The overall architecture test system 3.2 The basic system implementation process

Testing process is a real cell phone users "inputcondition-action rules(ECA-RULE)

Control Agent This is a test based on the core rules, which defines a test case of interest, and according to comparison conditions, take appropriate action.⑵ exception handling rules(EM-RULE)

Test Control Agent also comply with exception handling rules, which occurred during the test can detect abnormal events and to implement appropriate solutions to enable testing to proceed smoothly.3.4.2 Test Control Agent's Properties ⑴ autonomy

Autonomy to make the tester without too much attention to the details of the test, testers just need to select the test objectives, test control Agent information can be based on the knowledge base, automatically generate ECA rules table, and run the script, according to ECA rules to capture sensitive event and perform the appropriate action.⑵ intelligence

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Test Control Agent to the test objectives based on user selection, automatically generates ECA rules table, and according to ECA rules, reasoning and take appropriate action.Test Control Agent can exception handling rules based on the testing process to deal with abnormal body fat to protect the testing process automatically and smoothly.3.5 Agent Network

Network Agent implements the network environment simulation and control.In order to test the phone's RF performance, usually the base station simulator to provide a device in which the wireless network environment, and use programming interface, the software environment to control the device, making the complex network environment, the target device under test as possible.4 System Evaluation

In order to evaluate the effectiveness of the system, we designed a comparative experiment.Tell testing laboratory in the Ministry of Information Industry of the test project for real.We will test engineers divided into two groups, the first group to use this system MobileTest test, the second group uses the industry's leading TestQuest Pro test.Test is based on the number of China's telecommunications industry standard part of mobile station testing.Test content is based on both systems return to their already established on the new test cases for regression testing of smart phones to compare these two test tools, test efficiency and maintainability of the script.Table 1 regression test results:

Two experimental tools to measure different types of test cases in the test, the automated implementation of the percentage of use cases.Can be seen from the table, although the functional testing TestQuest higher automation rates, the stress test, multi-state testing, multi-tasking tests, critical test, and the total, MobileTest better test coverage.This is because after the four tests, the test case need to constantly track the system state, the use of pictures than on the mechanism of TestQuest difficult to accomplish these tasks, and MobileTest able to use the Agent to return to the treatment of sensitive events.In addition, MobileTest time to complete the test task 15 days, TestQuest's test efficiency than higher.专业英语期末考查

第13页 Conclusions

This paper presents a technique based on embedded intelligent devices Agent test method, which uses embedded intelligent Agent technology equipment is tested in test equipment, test and simulate and control the network environment, a good shielding of the embedded the complexity of smart devices to improve the test efficiency and reusability of the script.Future research work, we will further develop the test system so that it can support the whole testing life cycle.The innovation of the author: This article introduces the software Agent technology to automate testing of embedded intelligent devices, the use of Agent's features, nice screen test for embedded smart devices, testers, test device, the complexity of the network environment and increase the efficiency of automated testing.From:《 Journal of Electronic Science and Technology》

第五篇：专业英语论文翻译

MET基因复制数量的增加赋予单克隆抵抗体抗MET的能力并且建立药物依赖性

关键词：MET，MV-NV30单克隆抗体，酪氨酸激酶抑制剂，抗性，药物依赖性 【摘要】：被MEI原癌基因编码的酪氨酸激酶受体领导了具体抑制剂的发展并在癌症中起很重要的作用，其中现在一些正处于前进的临床试验阶段就以前的经验表明对大多数靶向治疗最主要的限制是抗性的出现。在对MET单克隆抗体抗性和抗体对化学抑制剂旁路抗性（反之亦然）一无所知时，酪氨酸激酶抑制剂对MET的抗性机制就已经被提出。EBC1型肺癌细胞是MET基因扩增的结果，并且这种细胞对MET抑制剂非常敏感，包括MET单克隆抗体的单机形式在内。我们培养生成抵抗抗体的细胞发现这种抗性是由于MET基因大量复制扩增和它的受体显著表达而来。这种过度表达可以使单克隆抗体的“脱落”活动达到饱和，并且能够防止表面的MET受体的有效的下调和抑制剂的活化作用。值得注意的是MV-DN30抗体的抗性细胞是MET耐受细胞对MET酪氨酸激酶抑制剂也很敏感。除此之外，抗体抗性细胞还具有药物依赖性,MV-DN30的去处导致它们死亡是由于它们的过度信号表达。在实验中，对MET酪氨酸激酶抑制剂存在抗性的细胞仍然对MV-DN30抗体的作用敏感。结果表明一种不连续的通过抗体和化学激酶抑制剂联合治疗可能会使靶向治疗的临床反应和对MET抗体治疗旁路的抗性增加。

缩略语：MV-DN30--单价DN30，TKEs—酪氨酸激酶抑制剂，HGF—肝细胞生长因子，MAPK—有丝分裂原活化蛋白激酶 1.简介

可以抑制一个特定的目标分子化合物的靶向治疗法开辟了治疗癌症的新道路。与主要杀死扩散细胞为主的传统化疗不同的是靶向药物对肿瘤细胞采取一种更具体的治疗方式。靶向治疗依赖于“癌基因沉瘾”的概念。这就意味着单个基因的抑制或死亡是由于它们的沉瘾，或者至少抑制它们的生长（温斯坦,2002）。临床试验中的特定抑制剂的发展给肿瘤细胞的“Achille’s heel”的识别提供支持（温斯坦和乔，2006）。

尽管靶向治疗在一部分癌症患者中取得了较优异的效果，还有重要的一点就是部分癌症病人对药物的选择表达没有起到治疗作用（原发性），除此之外，几乎总是一开始患者反应变成后来对治疗的抵抗和复发（继发性）。因此，最关键的就是要发现对治疗抵抗的机制并且找到绕过它们的方法。

癌基因和人类癌症密切相关，酪氨酸激酶起着决定性作用。这个观察发现许多肿瘤沉溺其中使得蛋白激酶成为了治疗癌症的理想目标（巴塞尔加，2006;Gschwind等人，2004）。在临床诊断中主要使用一种较小的激酶抑制剂和单克隆抗体来抑制酪氨酸激酶。酪氨酸激酶抑制剂是一种可以抑制靶蛋白酶活性的小分子物质。它们能有效地瞄准膜结合位点和细胞内的激酶并且很容易在体内扩散。单克隆抗体已被广泛用于临床并取得了可观的成果。这些分子的优点在于它们具有很高的特异性。在癌症治疗中的可以抗癌的RTKs单克隆抗体已经被批准在乳腺癌和结肠癌中使用(分别针对HER2和表皮生长因子受体)并可作为抗血管增生的药物（针对血管内皮生长因子受体）。除此之外，很多针对于其他目标的单克隆抗体正处与发展和试验阶段。最近，一种作为癌症治疗目标的RTK受到关注，这种RTK是由致癌基因编码的在肝细胞生长因子上的酪氨酸激酶受体。在和肝细胞生长因子结合后，MET活化并启动一个复杂的生化程序，这个过程被称作“浸润性上长”。在肿瘤组织中，浸润性生长的增进可以迫使肿瘤细胞从肿瘤组织中分解下来侵蚀基底膜，渗入基质中，甚至定居于新的组织中来实现转移。很多研究结果表明MET在人类的许多肿瘤中具有活性，并且它与对直接激酶疗法的持续抗性密切相关。除此之外，还表明细胞显示大量复制（超过8张）和随之而来的过度表达和独立配体的激活都是沉溺于这种致癌基因和抗MET药物的应答中。在前期设置得到的基本结果，几种特定的多目标的酪氨酸激酶抑制剂和直接针对于MET或者HGF的抗体已经进入了临床试验阶段。在活体和动物模型进行的研究已经表明用TKIs长期治疗会导致机体的治疗耐受性。对MET TKIs的抗性可能是由于一些机制，比如MET基因扩增，过度表达，MET点突变，MET平行路径的激活和KRAS基因的扩增机制。然而，关于对MET的单克隆抗体的再次具有抗性一无所知。

我们以前报道过主要针对细胞外的部分MET的抑制性单克隆抗体的研究进展。它的诱导、再结合、达到MET脱落阈值的能力使其有抑制活性，剩余的跨膜片段通过蛋白酶体降解途径处理掉。因此，DN-30结合到MET后的结果是使其变成可溶性的诱饵MET并且蛋白水解酶会讲解MET激酶。这促进了MET介导的生物活性的抑制作用。设计了这样一个过程就是因为DN-30的结合使得MET激酶部分活化并且导致抗体介导的受体同源二聚体化和单价Fab片段失去竞争活性的一个过程。

在这个研究中我们表明了不断用MV-DN30来治疗沉瘾癌细胞会使其具有抗性的原因是MET基因的大量复制和MET的过度表达超过了MV-DN30对其有效下调并使其失去活性的的能力。值得注意的是，MV-DN30抗性细胞还会一直对MET TKIs产生耐受性和敏感性。有趣的是，它们获得了药物耐受性，当受到MV-DN30的驱除致死它们的是过多的信号表达。我们还表明对MET TKIs 有抗性的细胞也对MV-DN30敏感，所以，MV-DN30和MET TKIs 对肿瘤细胞的作用是相互促进的。2.材料与方法 2.1.细胞和试剂

EBC1 细胞从一个患有转移皮肤肿瘤的病人取得，这个患者还患有肺鳞状细胞癌，病例是从日本癌症资料库购买得到。GTL16 是一种实验室里的克隆胃癌细胞系。HEK-293T细胞系分离于人类胚胎时期的肾，A549细胞系来源于肺癌，都是从ATCC购买来培养的。对MV-DN30有抗性的EBC1细胞可以通过一个逐步的方法培养得到，由Sigma Tau R&D 提供的通过暴露亲代细胞方法来增加抗MET单价单克隆抗体的浓度。亲代细胞用10 mg/ml 的MV-DN30治疗约一个月，直到生成的R10细胞开始产生抗性为止，R10抗性细胞又用逐步增加浓度的MV-DN30治疗。所有的抗体耐受细胞培养在存在MV-DN30并且可以使它们产生耐受性的条件下。大约两个月可以分离到R20抗体抗性细胞，四个月可以分离到R80抗体耐受细胞。EBC1 and GTL16 细胞对MET TKIs PHA-665752(EBC1 RPHA 50 nM and GTL16 RPHA 150 nM)都有耐受性，并且向描述的那样培养可以一直保持PHA-665752的存在。该细胞系的遗传身份通过一个短的串珠状重复序列（STR）识别，这段序列在2013年7月再次重复出现。We 我们利用下面的小分子：ATP竞争行MET TKIs PHA-665752(Tocris Bioscience)and JNJ-38877605(John-son & Johnson)和p38MAP激酶抑制剂SB203580(Merck).2.2.mRNA和基因组DNA的分析

用Trizol 试剂提取得到的RNA被检测到利用多文士病毒的逆转录酶和随机引物合成，cDNA可以用实时的利用电源带动的绿色PCR混合的PCR 技术来实现扩增，根据制造商的说明下面的MET和ACTIN特异性引物要用到： hMET ex 19 Fw: 50-AGTTTACCACCAAGTCAGATGTGT-30;hMET ex 20 Rw: 50-GGGCTCCTCTTGTCATCAGC-3;hACTIN Fw: 50-GGAGGAGCTGGAAGCAGCC-30;hACTIN Rw: 50-GCTGTGCTACGTCGCCCTG-30.根据制造商的说明用实时PCR技术来分析用纯化的DNA基因组迷你试剂盒分析从细胞中提取到的基因组DNA，这种技术是用 TaqMan基因表达的主要结构和TaqMan探针MET基因和RNaseP控制基因的实时定量PCR分析。MET的mRNA的成倍增加和EBC1中MET基因的大量复制以及MV-DN30抗性细胞归一化然后被认可。2.3.蛋白印迹分析和脉冲追踪代谢标记

蛋白提取液(40 mg), 细胞上清液(20 ml)。在细胞裂解前2小时把MET TKI JNJ-38877605加入到指定的地方。免疫印迹法使用了以下的初级抗体：the anti-MET Intracellular domain(ICD)(zymed, #370100)from Invitrogen, anti-MET ECD(DL21)obtained as described(Prat et al., 1991), anti-phospho-Tyr1234-Tyr 1235MET(#3126), anti-AKT(#9272), anti-phospho-Ser473AKT(#4060),anti-p44/42MAPK(#9102),anti-phospho-Thr202-Tyr204p44/42MAPK#9101),anti-p38MAPK(#8690),anti-phospho-Thr180-Tyr182-p38 MAPK(#9215), from CellSignaling;anti-vinculin(#V9131)from Sigma and anti-b-actin(#I-19 sc-1616)from Santa Cruz Biotechnology.Secondary IgG HRP-Peroxidase antibodies were from Amersham.脉冲追踪实验的1106WT，或MV-DN30抗性的EBC1细胞(R80)都被平铺在60mm的盘里。R80 细胞保存在有或没有抗体（80 mg/ml）存在的条件下，而WT细胞要一直保存在有抗体的条件小16小时。然后，这些细胞在不含L-蛋氨酸但有500mmMCIL-甲硫氨酸S35(脉冲)（易标记）的DMEM培养基中处理20分钟。在这之后，去处放射性标记的培养基，细胞用1ml磷酸盐缓冲液盐水洗两次然后保存在有2mlISCOVE的培养基中，培养基中加入2%FBS，在MV-DN30(80 mg/ml)存在或不存在脉冲3.6和16小时。之后，用免疫沉淀法（IP）在1ml细胞裂解液中进行测定（裂解缓冲液：1% TritonX-100存在下，20 mM Tris-盐酸，5毫米EDTA，10% V / v甘油，150 mM NaCl补充蛋白磷酸酶抑制剂）通过使用抗Met ICD dq13单克隆抗体，而IP法利用抗Met ECD do24单克隆抗体对细胞培养2毫升上清液进行。细胞裂解液和上清液都在有已知抗体的培养基里培养16个小时，抗鼠IgG抗体预包被琼脂糖凝胶蛋白珠形成免疫复合物沉淀下来。免疫沉淀物中的蛋白就会被8% SDS-PAGE 分开，然后转移到3mm的纸上，80度，48小时后蛋白质的放射性就会在投影膜上留下印记。2.4.生长和可行性分析

用于细胞生长和可行性分析的这些细胞被接种在96孔的培养板上，根据制造商的说明用已知药物在不同时期进行处理然后用细胞滴度发光细胞进行可行性分析。没经过处理的细胞控制在药物载体存在的条件下生长。所有的数据进行归一化到0天的药物治疗。2.5.荧光细胞分析

对结合在EBC1WT,R20,R80细胞质膜上的MET进行免疫荧光着色，这些细胞要提前在有或没有MV-DN30存在的条件下培养24小时。荧光性的强度可以通过细胞荧光性分析检测到。用于检测的细胞要在PBS中用2%FBS洗涤并且在室温下用抗MET ECD DO24 mAb(100 ng/ml)着色20分钟。然后在室温下细胞又在PBS中用2%FBS洗涤就会逐步产生抗鼠IgG-RPE二抗然后用二脒基苯基吲哚作用20分钟。作为阴性对照，将不含初级抗MET抗体的细胞进行染色。质膜结合的蛋氨酸的荧光强度（AU为单位），通过使用GraphPad Prism软件绘制为箱形绘图图表。

2.6.慢病毒载体转导

EBC1 WT 细胞能稳定地在两种不同量的慢性病毒颗粒编码的MET cDN转导，包括1mg(METþþ)和1.6mg(METþþþ)的p24病毒抗原，其浓度按说明确定。作为对照，WT细胞用含有空载体病毒颗粒感染。MV-DN30抗性细胞(R20 and R80)只用空载体感染。在感染48小时后接种细胞用于生物化学分析。转导细胞的可行性分析如先前描述的一样进行，让细胞在存在或不存在MV-DN30的条件下上长72小时。如先前所述，MET的蛋白印迹分析和磷酸化的MET蛋白水平和对感染细胞的mRNA的表达水平实时定量PCR分析一样子细胞感染72小时后进行评估。2.7.药物协同作用分析

在抗MET抗体MV-DN30和MET TKI JNJ-38877605之间进行药物协同作用分析是对WT EBC1和接种在96孔板上的GTL16细胞在用药物治疗72小时后的细胞活力的研究。如果使用一个药品从药物剂量来说要比混合使用的IC50约高10倍左右，双重增加浓度可以用于单药使用和组合使用。评估细胞活力为前面描述的增效作用的药物效应多种分析研究，采用组合指数（CI）和Chou and Talalay 方法。使用相互排他性假设计算CI值（药物作用机理的不同，采用compusyn.exe）软件，可上线的网站：http://compusyn.software.informer.com/，和绘制功能的FA（由这两种药物的组合影响系统分数）。CI值< 1表明两种药物之间的协同作用。2.8.统计分析

同一个实验数据用GraphPad Prism软件进行两尾t检验分析至少有三种生物学具有统计学显著意义的结果。P值小于0.05被认为是有意义的。3.结果

3.1.MV-DN30耐药细胞株的建立

EBC1 肺癌细胞是MET沉瘾细胞，它能放大MET的扩增、过度表达和活化过程。MET TKIs或者MV-DN30对EBC1细胞里的MET具有强烈的破坏它们生存和上长的能力的作用（图A~C）。

我们先前培养的EBC1细胞 对不同的YKIs有抗性表明了这种抗性可能是由于MET基因的进一步扩增。为了产生抗MV-DN30的细胞，我们采用逐步暴露EBC1原代细胞不方法来增加抗体的浓度，并且获得抗不同剂量抗体的细胞系（图1b；约的分步方法详见材料和方法部分）。, 进一步的实验中，我们使用耐20细胞（ebc1 R20）和80（ebc1 R80）毫克/毫升mv-dn30，剂量是分别是前面的约10倍和40倍，然后测这些细胞的抗体IC50（图1a）。在有MV-DN30存在的抗性细胞的生长速度一直和WT细胞差不多（图1C）。

正如前面提到的，MV-DN30发挥它的抑制活性是通过诱导MET的结构域蛋白反正溶蛋白裂解，随后又通过蛋白酶体受体介导受体降解。这种反应减少了细胞表面的MET的大量表达，抑制了MET的活性和受体介导的生物学活性。如图1D所示，事实上，MV-DN30的治疗导致了再EBC1细胞中的MET通过酪氨酸酶的磷酸化消除大量减少。与抗性细胞的生化分析相反的是显示了在相同剂量抗体存在的情况下，MET也会磷酸化并且在质膜上的大量的MET蛋白明显高于保存在相同条件下的WT细胞（图1D,E）。除此，尽管激活的下游目标AKT和MAPK激酶都被保存在有抗体存在的条件下（图1D）。

所有的数据表明，抗EBC1细胞的MV-DN30并不是通过一种足以消除它的结构的活性的方法来下调MET，而是一种允许它的信号转导来调节它的结构磷酸化的方法.3.2.抗性细胞中缺乏MV-DN30抑制活性并不是因为它的活性不足

如图1D,E所示，用MV-DN30处理抗性细胞致使MET大量减少这种减少比处于同等条件下观察的WT细胞中的减少更明显。我们想知道在抗性细胞里的抗体受损是否是由于MET的结构域IPT4发生突变引起，这个结构域是它的集合位点，但是我们没有发现任何突变（数据显示）。

于是，我们评估了是否是抗性细胞中脱落的MET胞外结构域（ECD）对抗体的反应被抑制了。在相同剂量MV-DN30存在的条件下培养了24小时的抗性细胞和WT细胞的上清液中，抗性细胞上清液中含有更丰富的脱落的ECD（图2A）。当对去处抗体24小时后的抗性细胞中MET的表达和活化进行评估时，我们观察到MET总量强烈增加特别是结合在细胞膜上的形式（图2B和图1）。对抗性细胞再引入MV-DN30细胞会导致上清液中MET ECD含量更高，与在同样剂量处理的WT细胞相比，之后细胞内的MET同时减少了并返回到平时观察的抗性细胞的一半水平（图2 D）。在脉冲代谢标记实验中也观察到了相似的结果：比起WT细胞，抗性细胞合成更多的MET和MV-DN30来促进MET ECD 释放并大量增加（图2E）。最后，用更高剂量抗体20 mg/ml(R20)处理EBC1细胞来更好的证明MV-DN30在抗性细胞中是具有活性的。图2F所示在抗性细胞中随着MV-DN30剂量是增加减少了大量MET在细胞膜上的表达（图2A）和磷酸化(图2B)。

总之，所有的结果表明EBC1细胞的抗性不是因为MV-DN30活性缺乏，而是由于用它的饱和能力来促进了一种高效的MET解离。

图1-抗MV-DN30的EBC1细胞分子表征。(A)在有MV-DN30的递增浓度的条件下培养了72小时的EBC1 WT细胞的细胞活力。对未经过处理的细胞(100%)± s.d.的归一化结果。(B)在指定浓度抗体的条件下培养72小时的EBC1 WT细胞或产生MV-DN30抗性的细胞(R10, R20, R40, R80)。与未经过处理的WT细胞(100%)± s.d归一化结果。(C)在MV-DN30(20 and 80 mg/ml)存在的条件下生长的EBC1 WT细胞EBC1R20和R80细胞的生长活性。与未经过处理的WT细胞(100%)± s.d归一化结果(***P < 0.001)。(D)在有指定剂量MV-DN30抗体存在的条件下处理24小时的EBC1WT ,EBC1R20和R80细胞的蛋白印迹。(E)结合MET的在24小时在不存在或存在的血浆膜的荧光强度的箱图mv-dn30 R20和R80生长EBC1 WT细胞（AU：任意单位）

图2—细胞耐受性并不是由于失去了对MV-DN30的敏感性。(A)在存在或不存在MV-DN30的环境下培养24小时的EBC1 WTR20和R80细胞的上清液中MET ECD的蛋白印迹。(B)在有或没有（24小时去除）的EBC1WTR20和R80细胞中的蛋白（顶部）和蛋氨酸的酪氨酸磷酸化蛋白印迹。纽蛋白作为控制基础(C)在 EBC1 WT, R20 and R80 细胞中有或没有（24小时去除）MV-DN30的条件下在质膜上结合的MET的荧光强度的箱图。质膜着色如图E。(D)在用MV-DN30处理（24h）的EBC1WT R20和R80 细胞中和抗性细胞（图2 D）中观察到的MET总蛋白水平和MET ECD 的蛋白印迹.(E)在有或没有MV-DN30存在下的EBC1WT和R80细胞里的MET(来源细胞裂解液，上面)和MET ECD（来源与细胞上清液，底部）的用脉冲追踪代谢蛋白标记法的免疫沉淀分析。(F)在递增MV-DN30浓度的环境下的EBC1(左图)和R20细胞（右图）的活性检测。未经过处理的WT细胞（左图）或者用20 mg/ml(100%)处理的R20（右图）统计结果(***P < 0.001, *P < 0.05)。3.3.对MV-DN30的抗性是由于MET的扩增和过度表达

如上图所示（图2BeD），MV-DN30抗性细胞表达MET水平比原代细胞更高。我们怀疑这可能是由于增加的启动子活性或更高的mRNA的可用性，因为你微RNA的负控制微分表达了。在EBC1WT和抗性细胞的荧光素酶检测结果显示排除这些可能性（数据未显示）。

然后我们评估了基因扩增的情况是因为MET大量复制是对激酶抑制剂存在抗性的机制。如图3所示，EBC1抗性细胞显示了MET的大量复制（原代细胞的第24代到30代），这大约是二倍体细胞的15倍。当分析MET的mRNA水平时，我们观察到MET的过度表达是WT细胞的2~3倍，达到了MET正常表达细胞(A549 and HEK-293T)的60~90倍。为了证明这种增加可能是为了维持对MV-DN30的抗性，我们将EBC1WT细胞放在不同量的MET cDNA条件下转导，通过比较EBC1R20和R80细胞mRNA的表达水平来获得（图3）。我们观察到，MET的mRNA以2~3倍的速度剧增，这与在存在MV-DN30的条件下细胞的活性有关（图3D）。除此，尽管有MV-DN30的存在MET保持磷酸化，这就解释了为什么细胞能够存活增加（图3E）。

所有实验结果表明，嗜MET的EBC1细胞变得有抗性是因为MET的进一步扩增和过度表达从而来阻止有效的抗体介导的对MET活性的抑制。图3

3.4.抗MV-DN30的细胞也是嗜MET细胞并有药物依赖性

为了证明抗MV-DN30的细胞也是嗜MET细胞，我们用一种小分子的激酶抑制剂JNJ-38877605来处理细胞。如图4a,b所示，抗MV-DN30的EBC1细胞在有抗体(20 mg/ml for R20 and 80 mg/ml for R80)存在的条件下生长情况显示了当用MET激酶抑制剂JNJ-38877605(10 nM or 250 nM)处理时，细胞活性的降低和MET磷酸化作用的减弱。和亲代EBC1细胞相比，抗性细胞对JNJ-38877605的敏感剂量更低，可能是由于在抗性细胞中MET的蛋白合成和转运更高效。从抗性细胞的培养基中去除MV-DN30会导致MET的表达增加（图2B），我们研究了过度表达的生化效应。抗性细胞培养在不存在MV-DN30的条件下，显示了活性下降（图4C）。这些细胞的蛋白印迹分析表明，随着时间的推移逐渐增加了磷酸化（图4D）。然而，24~48h后，after 24e48 h, p38 MAPK的活化清晰可见证明了通常在细胞凋亡反应之前会有一个细胞应激反应。除此之外，在没有MV-DN30存在的条件下培养的抗性细胞中的p38 MAPK会受到一种小分子SB203580 的抑制使得细胞活性得以恢复（图3）。这些数据进一步表明，p38 MAPK信号对于参与药物依赖是至关重要的.重要的是要记住对于嗜MET细胞抗MET酪氨酸激酶抑制剂的药物依赖条件和黑色素瘤有获得性的药物依赖性已经被证明。为了证明当去除MV-DN30后过多的MET信号使细胞凋亡是正确的，我们用低剂量的MET YKI处理抗性细胞让其充分减少但不会完全抑制MET活性（图4）。事实上，如图4E所示，保存在不含MV-DN30并用 10 nM JNJ-38877605（低于IC50浓度）处理的抗性细胞可以恢复它们的活性，并且这种水平类似于保存在有抗体存在的培养条件下的抗性细胞的活性水平。

总之，这些结果表明，MV-DN30抗性细胞都是嗜MET细胞并且它们的繁殖和生存都有药物依赖性。图4

3.5.在嗜MET细胞中MV-DN30和MET TKIs的协同作用

MET-TKIs 和 MV-DN30 都可以有效的抑制EBC1细胞的活性，我们想知道这两种抗MET化合物是否表现增强或协同作用。单独或在有非常低剂量的MV-DN30存在的条件下（0.15~2.5毫克/毫升，从10倍以下的IC50开始），我们用逐步增加MET抑制剂JNJ-38877605的方法来处理EBC1 WT细胞。然后我们分析细胞活力和对药物治疗的两种药物的组合效应的性质，利用多药效果分析。如图5C所示，联合治疗导致的剂量尽可能低为1.25~20 nm jnj-38877605在0.15~2.5毫克/毫升mv-dn30存在活性降低。当在GTL16 WT 胃癌细胞上做这个实验室会得到相似的结果（图5D）。如图5A、B所示，多药效应分析表明了 EBC1 and GTL16 WT细胞的CI值都小于1，从而表明了JNJ-38877605和 MV-DN30之间有药物协同作用.所有的这些数据表明用MET TKI和MV-DN30联合治疗可以有效地减少嗜MET细胞的活性，剂量明显低于单独一种药物治疗的药量。图5

4.讨论

临床效应甚至是最有效的靶向治疗都被发展的耐药性所限制。显而易见，这种耐药性机制已经被广泛研究。从癌细胞和抗酪氨酸激酶抑制剂表明，获得性耐药的最常见的机制包括在药物目标本身的二次突变的患者获得的数据，激活下游信号转导或平行的信号转导通路的激活突变。即使对关于癌症是怎样产生耐药性抗体的了解很少，一些现象提示用抗EGFR 和抗HER2抗体治疗的病人体内有耐药机制的建立。在EGER中的二次突变表明了免疫调节药物与EGFR的结合受到破坏从而调节了抵抗力。除此之外，最近的研究还证明了EGFR或HER2的信号转导通路的激活可以分别绕过西妥昔单抗或曲妥单抗的抑制；这些包括EGFR配体水平增加，MET RTK的扩增或过度表达，下游或平行激活信号转导途径。

在癌症治疗中MET受体已经变成了最具关注的目标，因为很多研究表明，MET在多种人类肿瘤中具有组成性活性。它的失调可能是由于不同的机制包括过度表达，基因扩增，激活突变和受体介导的刺激自分泌或旁分泌的增加。生殖细胞的激活突变的识别和遗传型肾乳头状癌直接证明了MET和人类肿瘤发生有关的概念。MET的酪氨酸激酶的结构域中激活突变位点已经在散发肿瘤中确定。然而，在人类癌症中变化频繁的是转录表达的过程，这个过程是由原癌基因激活诱导抑癌基因失活和对特定的为mRNA或缺氧刺激的下调过程。在原发性肿瘤中由基因扩增引起的MET过度表达是少见的，但在肺癌和结肠癌中变得抗靶向其它RTKs治疗的情况更频繁。临床证据表明癌细胞是沉溺在原癌基因中的细胞，在这种细胞中的MET具有组织性活性并且它们的抑制结果就是使其致瘤性被破坏。然而，用小剂量的激酶抑制剂延长对嗜MET细胞的处理会导致二倍抗性的出现，这种机制可以被维持比如MET扩增或突变,KRAS的扩增或EGFR家族成员的活化。在我们的研究中表明,在嗜MET细胞中对MET特定抗体MV-DN30的抗性获得是由于MET复制数量的大量增加。这一现象的机理解释依赖于染色体外的MET复制的动态调节，造成了一种癌症对治疗的适应性程序。MET上调的增加是由于MET基因扩增造成的这种现象并不明显当抗性细胞培养在有抗体（这种抗体介导了MET的下调）存在的条件下，但是在没有这种抗体存在的条件却是很明显清晰的。我们观察到蛋白的增加平行下来是METmRNA增加的2~3倍。EBC1 原代细胞的METcDNA的转导可以达到一个和抗性细胞一样是表达水平，证明了事实上细胞表达更多的MET可以减少对MV-DN30抑制活性的敏感性。WT EBC1细胞和大多数嗜MET细胞一样可以表达比正常表达细胞的约30倍多的MET；因此，在抗性细胞中2~3倍的增加可以引起MET水平比那些正常细胞中的MET水平高60~90倍。在缺氧的环境下观察A549细胞中MET呈3倍增加才能满足它的生化效应，在抗MV-DN30的EBC1细胞中MET增加可以解释为什么抗体能够有效地下调MET。除此，作为抗性细胞识别增加的脱落的MET导致了MET ECD在细胞内的累积，并且脱落的ECD可以和细胞膜上的MV-DND30结合位点结合从而减少抗体的抑制效应。抗性细胞仍然依赖于MEI的信号传导来增殖和生存，MET可以完全被TKIR抑制从而导致它们的增殖受到抑制。有趣的是，当MET变得非常活跃的时候，可以通过增加在MV-DN30存在下的MET 的表达在这种适宜的有利的环境下来使其对药物的去除效果减弱。我们的结果和报告是一致的都表明了，正常细胞和肿瘤细胞不仅对信号（即该通路被激活的信号）的质量敏感而且对数量也很敏感（这种量就是怎样激活通路）。信号过量可导致细胞应激（在我们的情况下，通过p38 MAPK活化）导致细胞死亡。事实上，在抗体缺乏，低剂量的遇到了TKI治疗耐药细胞（抑制野生型细胞活力）增加了他们的活力，减少了信号的水平细胞持续强度的可能。Das Thakur做了一个类似的观察，这个人证明了人类黑色素瘤移植提供的抗药性，药物的去除使耐药肿瘤的肿瘤消退是由于依赖BRAT信号过量。有趣的是，他还表明不连续的药物剂量，运用耐药细胞在没有药物环境下的劣势来防止耐药性的发作。这在嗜MET细胞中也被证明是正确的，无论是抗MV-DN30还是抗TKIs的EBC1细胞都表现出药物依赖性并且去除药物会致死细胞。因此，可以想象，为了防止由于癌基因过量用药而产生抗性，间断性用药的机制可能会比连续用药更有效。

此外，我们表明，抗体治疗是活跃的细胞提供的抗酪氨酸激酶抑制剂，相反，TKI治疗耐mv-dn30细胞有效。这并不奇怪，因为这两种类型的抗性细胞都是嗜MET细胞，因此，依赖MET的信号减少导致了它们的死亡。这种发现在临床上受到关注，因为它表明了用抗MET的抗体治疗病人可能是没用的，病人的对TKIs的抗性是由于MET表达的增加或者是它们的毒性限制了TKI的剂量。另一个处于我们的观察的结果是MV-DN30和MET-TKIs之间的协同作用。值得注意的是，尽管是几个实验，在用MET TKI和MV-DN30治疗时我们并没能过培养出抗性细胞。这些观察的临床意义提出了一点就是两种药物剂量的减少的可能性，结果它们又不利影响当需要保持治疗效应时有可能会降低或延迟。总的来说，在事实的角度评估我们的结果就是它们表明了两种可能会克服或防止对METmAbs抗性的治疗策略:(i)一种就是用抗MET的mAb加上MET TKI的联合治疗(ii)利用抗性细胞的药物依赖性的定期间断性的节律治疗。