A Strategy for Precise and Large-Scale Identification of Core Fucosylated Glycoproteins
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大水灌溉的英文作文英文:I think that irrigation is a very important practicefor agriculture, and large-scale irrigation can be very beneficial for farmers. With large-scale irrigation, farmers can water their crops more efficiently and effectively, which can lead to higher crop yields and more successful harvests.There are a few different methods that can be used for large-scale irrigation, including flood irrigation, sprinkler irrigation, and drip irrigation. Flood irrigation involves flooding a field with water, while sprinkler irrigation involves spraying water onto the crops from above. Drip irrigation, on the other hand, involves slowly dripping water directly onto the roots of the crops.Each of these methods has its own advantages and disadvantages. Flood irrigation can be very effective atwatering large areas quickly, but it can also lead to soil erosion and water waste. Sprinkler irrigation can be more precise and efficient, but it can also be more expensiveand require more maintenance. Drip irrigation is very efficient and precise, but it can also be more expensiveand require more installation time.Overall, I think that large-scale irrigation can be a very beneficial practice for farmers, as long as the right method is chosen and implemented properly. By using large-scale irrigation, farmers can ensure that their crops receive the water they need to grow and thrive, which can ultimately lead to more successful harvests and a more sustainable agriculture industry.中文:我认为灌溉是农业中非常重要的一项实践,大规模灌溉可以为农民带来很多好处。
空间计算英语Space calculation is an essential aspect of various fields, including architecture, interior design, engineering, and construction. It involves determining the area, volume, or dimensions of a given space, which is crucial for planning and designing purposes. In this article, we will discuss the importance of space calculation, its applications, and the different methods used for accurate measurements.Space calculation is important because it allows for efficient use of space and resources. In architecture and interior design, accurate measurements are crucial for creating functional and aesthetically pleasing spaces. Engineers and construction professionals also rely on space calculation to ensure that structures and buildings are safe, stable, and meet regulatory requirements.One of the most common applications of space calculation is in floor area measurement. This is particularly important in real estate, where the accurate measurement of floor area directly impacts property valuation and sales. In addition, space calculation is used in determining thecapacity of a room or building, which is essential for event planning, occupancy load calculations, and safety considerations.There are several methods used for space calculation, depending on the type of space and the level of accuracy required. One of the most basic methods is manual measurement using a tape measure, ruler, or laser distance meter. This method is suitable for small-scale projects or quick estimations but may not be precise enough for large or complex spaces.For more accurate measurements, advanced tools and techniques are used, such as computer-aided design (CAD) software, 3D scanning, and laser scanning. These technologies allow for precise and detailed measurements of spaces, including irregular shapes and contours. They are often used in architectural and engineering projects, where accuracy is critical for the success of the design and construction process.In addition to traditional methods, space calculation can also be done using mathematical formulas and equations. For example, the area of a rectangular space can becalculated using the formula A = l × w, where A is the area, l is the length, and w is the width. Similarly, the volume of a three-dimensional space can be calculated using the formula V = l × w × h, where V is the volume and h is the height.In conclusion, space calculation is a fundamental aspect of various industries and plays a crucial role in planning, designing, and constructing spaces. Accurate measurements are essential for ensuring safety, functionality, and efficiency in the use of space. With the advancement of technology and the availability of advanced tools, space calculation has become more precise and reliable,benefiting professionals in architecture, engineering, construction, and real estate.空间计算是各个领域的一个重要方面,包括建筑、室内设计、工程和建筑。
高中生物教材英语读音High School Biology Textbook English PronunciationBiology is a fascinating and complex field of study that explores the intricacies of living organisms. As a high school student, the biology textbook serves as a crucial resource for understanding the fundamental concepts and principles that govern the natural world. One aspect of the biology textbook that can be particularly challenging for many students is the English pronunciation of the scientific terminology and concepts.The English pronunciation of biological terms can be a daunting task for high school students as many of these words have their origins in Latin or Greek. The unfamiliar syllables and combinations of letters can make it difficult for students to accurately pronounce and remember the terms. This can lead to confusion and difficulties in comprehending the material presented in the textbook.One of the primary reasons for the complexity of biological terminology is the need for precise and unambiguous language. Biologists and scientists must use specific terms to describe the various structures, functions, and processes that occur in livingorganisms. This precision is essential for effective communication within the scientific community and for the accurate documentation of research findings.However, this precision can also create a barrier for high school students who are just beginning to explore the world of biology. The sheer volume of new vocabulary can be overwhelming, and the unique pronunciations can make it challenging to effectively communicate and participate in classroom discussions.To overcome this challenge, high school students must develop strategies for mastering the English pronunciation of biological terminology. One effective approach is to familiarize themselves with the common roots, prefixes, and suffixes that are used in the formation of these terms. By understanding the etymological origins of the words, students can begin to recognize patterns and make educated guesses about the pronunciation of unfamiliar terms.Another useful strategy is to practice the pronunciation of key terms repeatedly, both individually and in the context of the textbook material. This repetition helps to cement the correct pronunciation in the student's memory and can also aid in the retention of the conceptual knowledge associated with the terms.Additionally, high school biology teachers can play a crucial role insupporting students in their efforts to master the English pronunciation of biological terminology. Educators can incorporate pronunciation practice into their lesson plans, provide resources for self-study, and encourage students to ask questions and seek clarification whenever they encounter a term they are unsure how to pronounce.By developing effective strategies for mastering the English pronunciation of biological terms, high school students can gain a deeper understanding of the concepts presented in their biology textbooks. This, in turn, can lead to greater academic success and a stronger foundation for future studies in the field of biology.In conclusion, the English pronunciation of biological terminology is a significant challenge for many high school students, but it is one that can be overcome through dedicated practice, strategic learning techniques, and the support of knowledgeable educators. By embracing this challenge, students can unlock a deeper appreciation for the wonders of the natural world and pave the way for a successful and rewarding journey in the field of biology.。
Software testing life cycle The trend of testing is changing and updating day by day. Testers are now learning and using more techniques. Testing is not just finding a few bugs and error in the software it has a more extensive range and scope. Testing is a process which we follow after the development of software to get ensure whether the software is bug-free or not.Software testing is the most critical process in the creation of the software life cycle. Testing also possesses its testing life cycle, so the need for the testing starts from the starting of theprojects. Software testing life cycle follows the same process steps involved in the software development life cycle.As we know before the development of any software, it follows the software development life cycle. In the same manner, the testing phase of software development also supports the software testing life cycle. In this article, we will study the software testing life cycle in- depth and you will get to know about every phase of software development life cycle accurately.Lifecycle: let’s understandSequences of changes from one to another known as a life cycle. These changes can happen at any phase of development. The software testing life cycle is almost similar to the software development life cycle. As we all know software development life cycle involves some steps in sequences. Testing also involves some steps that must be executed in a proper way. The purpose of software testing is the execution of testing activities in such a way so that it could make the software bug free. Hence we can say life cycle is a process of change that can occur at any instance of point in the process of anydevelopment. It is a planned way of executing the entities involved in the life cycleWhat is Software Testing Life Cycle and Why is Testing important?Before talking about the software testing life cycle now, we will talk about why testing is essential?Testing is quite important because with the help of testing we check the whole process and try to find out those potential which can break down the process of any activity.1-It detects the issues and errors that can arise during the software development life cycle. And helps to find out the mistake made by the programmer during the implementation of the software2-It helps to maintain the Quality of the product. If we provide a quality product, it will help us to gain customer confidence.3-Help to find out the bug that can stop the process of development. And we can fix them easily4-It’s essential to ens ure that there should be no failure after the application development Because it can increase the cost, in the future or the later stages of the event.Now we know about the testing and its benefits, let’s talk about the software testing life cycle.Software testing life cycle is a process that involves the execution of specific steps in such a systematic way so that it could give us the surety that after the complete implementation of the process it will provide us desired result. In the software testing life cycle, we make proper planning and precise execution for every activityso that it could perform its desired task. Software testing life cycle has many phases, and every phase has different goals and objective are set.Here are some phases involved in the software testing life cycleRequirement PhaseIn this phase of the software testing life cycle, we examine and study the need for the process. And we arrange a brainstorming session with the other team member and talk about the requirements involved in the whole process,whether they are examinable or not. The primary purpose of this phase is to find the scope of the testing, and if any entity can’t be tested, we take a backup for this situation and make some strategy so that we could overcome such situations.Planning PhasePractically, we can consider the planning phase as the first phase of software testing life cycle. In this phase, we find out the resources and activities, which help us to reach the desired goal. The planning phase is a way of identifying, gathering and tracking the metrics. Wheneverwe do the planning, we should care about all the resources involved in it and. With the help of this, we make a proper strategy for the rest of the process.The planning phase involves two essential factors.Test strategy Risk analysis and mitigationWhat is Test Strategy?Test Strategy means using available resources at one’s disposal and uses them in such a manner so that it could achieve the desired goaleffectively. Test strategy is part of the planning, and it ensures that how the testing objectives would be met adequately to the desired expectations.Here are some advantages of a practical test strategy are;It reduces the possibility of risks which may arise during the STLC.we can focus and analyze the different aspect of the system under the testwe can clear the clarity of the requirements such as people, procedures, tools and infrastructure.Risk analysisRisk analysis is a process of identifying those issues which can make a negative impact on the whole process of development. It could be related to any process either it is software testing life cycle or any business life cycle.With the help of risk analysis, we minimise or avoid the possibility of those events which can affect the whole process and cause an error. It is a process of preparing ourselves for thoseevents which can occur while the development or testing the software.organizations use risk analysis to prevent those events which can be harmful to the process. It helps to minimise the probability of failureBenefits of risk analysis1-Help to identify the overall impact of the risk on the whole process and provide exact details of issues that can arise at any instance of time during the process.2-Improve the overall process.3-Steps in the risk analysis processThere are few steps involved in the risk analysis are given belowConduct a risk management surveyFirst of all, we conduct the risk management survey, and we work on those potentials which can affect the whole body of the process. We talk about those potentials and make a backup plan for them.Identify the risks:After conducting the risk management survey now the second step is to identify the threat and its potential. Once the risk is identified, we move towards the next stepAnalyze the risksOnce the threat is identified we analyse what is kind of risk it is? And what issues it can arise in the whole process.Develop a risk management planafter examining the entire process we develop the risk management plan and work on it in sucha way so that it could be implemented during process failure.Implement the risk management planif we face any issues while the software testing life cycle we implement the backup plan which we made while the risk management processMonitor the risks:<p style="text-align:left"> After implementation, we keep tracking the potential which caused an error in the whole process and makes sure that issue or risk should not arise again.Analysis Phase:In this phase, we identify what could be tested in the software testing life cycle. And behalf of this we perform the analysis process.There are many things which affect the test conditions on behalf of its identification.Product/project risksThe complexity of the productLevels and depth of testingAvailability of the stakeholders.Software development life cycle involved.Test management Skills and knowledge of the team.Design Phase:This phase involves many tasks and defines how to test, the test conditions, and in this phase, we divide the test conditions into the sub-conditions by which we increase the coverage.Identify test data.Identify and create a test environment.Make the requirement traceability metrics.make the test coverage metrics.Implementation Phase:The implementation phase involves the creation of individual test cases. In this phase, we assign the priority level to the test cases and identify which test case will take part in the reversion suite. It is quite necessary to review the test case before finalizing it. And take the accomplishment of the test case before the actual implementation takes place. In case your project consist of automation, identify thecandidate test case for automation and then we should move further for proper scripting of the test case. But the thing is you must not forget to review them.Execution Phase:The execution of all entities in the process takes part in this phase. But the most important thing is before starting the performance make sure that every object is met. Implement the test cases, and simultaneously fill your traceability by which you could trace the whole process.Conclusion Phase:Finally, in this phase of the software testing life cycle, we perform the final process by reporting and making the database. It is necessary because its a requirement of your project and also it depends on stakeholders choice we create a report and ask the stakeholders whether they want monthly or daily reports. Making a report for every single phase and requirements is quite an important thing. In the conclusion phase, we estimate our expenses and review the whole process.Closure Phase:It is the ending phase of software testing life cycle by which ensure whether the expectations are met or not. As we can see software testing life cycle is similar to the software development life cycle, but it is a lit bit more complicated. It involves all the phases involved in any software development life cycle and provides the essential detail of the development.ConclusionNow with the help of this article, we can understand the process of the software testing life cycle and know about the details of every single phase which get involved in the process.The method of software testing life cycle is almost the same as the software development life cycle but with the cycles are essential for the development of any software. Hence we can say we can get the initial idea of the life cycle and its functionality by learning this topic.。
模具相关英语词汇IntroductionIn the field of mold making and manufacturing, it is important to have a good understanding of the specialized terminology used in the industry. This document ms to provide a comprehensive list of mold-related vocabulary in English.Mold Types and ComponentsInjection MoldAn injection mold is a tool used in the manufacturing process of plastic parts. It consists of two primary components - the mold cavity and the mold core. The mold cavity is the space where the plastic material is injected and takes shape, while the mold core provides the shape and structure to the part.Blow MoldBlow molding is a manufacturing process used to produce hollow plastic parts, such as bottles and contners. The blow mold is a specialized tool that shapes and forms the molten plastic into the desired shape.Compression MoldCompression molding is a molding process that involves placing a preheated material into an open mold cavity, then closing the mold to apply pressure and heat. The compression mold is responsible for providing the desired shape and structure to the final product.Ejector PinsEjector pins are small rods or pins that are used to push the finished parts out of the mold cavity. They are typically located at the back of the mold and are activated when the mold opens.Runner SystemThe runner system is a network of channels that allow the molten plastic material to flow from the injection machine into the mold cavity. It consists of the sprue, runners, and gates, which control the flow and distribution of the plastic material.Cooling SystemThe cooling system in a mold is responsible for regulating the temperature of the mold and the solidified parts. It usually involves the use of water channels or cooling pipes to dissipate heat and ensure efficient production.Mold Release AgentA mold release agent is a substance or compound applied to the mold surface to prevent the material from sticking to the mold. It allows for easy removal of the finished parts.Mold Design and Manufacturing ProcessesMold DesignMold design is the process of creating a blueprint or layout for the mold, specifying the dimensions, features, and detls of the mold cavity and core. It involves considering factors such as material selection, part design, and production requirements.CAD (Computer-ded Design)CAD is a software tool used for creating 2D and 3D models of mold designs. It allows for precise and accurate design representation, as well as the simulation and analysis of mold behavior.CAM (Computer-ded Manufacturing)CAM is a software tool used for generating toolpaths and instructions for the machining and manufacturing of molds. It helps streamline and automate the production process.CNC MachiningCNC (Computer Numerical Control) machining is a manufacturing process that uses computer-controlled machines to remove material from a workpiece and create the desired shape. It is often used in mold making for precision and accuracy.EDM (Electrical Discharge Machining)EDM is a machining process that uses electrical discharges to remove material from a conductive workpiece. It is commonly used for intricate and complex shapes in mold making.PolishingPolishing is the process of using abrasive materials and compounds to create a smooth and glossy surface finish on the mold cavity and core. It is important to achieve a high-quality surface finish to ensure proper part ejection and minimize defects.Mold AssemblyMold assembly involves the process of fitting together the various mold components, such as the cavity, core, ejector pins, and cooling system. It requires precision and careful alignment to ensure proper functioning of the mold.Mold TestingMold testing is conducted to evaluate the performance and functionality of the mold. It may involve producing sample parts, checking for defects, and making necessary adjustments before full-scale production.ConclusionHaving a good understanding of the mold-related terminology is essential for effective communication and collaboration in the mold making and manufacturing industry. This document has provided acomprehensive list of mold-related vocabulary in English, covering various mold types, components, design, and manufacturing processes.。
英语作文80词英语对我的影响英文回答:English has played an instrumental role in shaping my life, becoming an integral part of my identity and expanding my horizons beyond measure. As a non-native speaker, learning English opened up a world of knowledge, communication, and connection.Through English literature, I have delved into the minds of literary giants, their words etching themselves into my consciousness. Their stories have taught me valuable life lessons, broadened my empathy, and kindled a deep appreciation for the power of language. Additionally, English has empowered me to express myself creatively, writing stories, poems, and essays that reflect my inner thoughts and feelings.Moreover, English has been my gateway to global communication. It has enabled me to connect with peoplefrom diverse cultures, forging friendships and fostering cross-cultural understanding. In academic and professional settings, English has served as a lingua franca, facilitating collaboration, knowledge sharing, and innovation on a global scale.Beyond its practical benefits, English has also enriched my life aesthetically. Its rich vocabulary and nuanced grammar have expanded my capacity for precise and evocative expression. I find joy in the rhythm and flow of the language, its ability to convey complex emotions and ideas with finesse.中文回答:英语对我的人生产生了深远的影响,它成为我身份中不可或缺的一部分,并极大地拓宽了我的视野。
PreciseIntroductionPrecision is an important concept in various fields such as science, engineering, mathematics, and even language. It refers to the level of accuracy or exactness in measurements, calculations, or descriptions. Being precise means being able to provide detailed and accurate information without any ambiguity or vagueness. In this article, we will explore the significance of precision in different domains and discuss its importance in various aspects of our lives.The Importance of Precision in Science1. Accurate MeasurementsOne of the fundamental requirements in scientific research is theability to make precise measurements. Accurate measurements are crucial for validating hypotheses, conducting experiments, and drawing reliable conclusions. Precision in measurement ensures that scientists can obtain consistent and reproducible results, which are essential for scientific progress.2. Precise CalculationsIn addition to accurate measurements, precise calculations are equally important in science. Precise calculations involve using appropriate mathematical models, algorithms, and formulas to analyze data and make predictions. By ensuring the precision of calculations, scientists can minimize errors and obtain more reliable and meaningful results.3. Clear and Precise CommunicationPrecision is also crucial in scientific communication. Researchers need to convey their findings, theories, and ideas in a clear and precise manner to ensure their work is accurately understood and can bereplicated by others. Precise language allows for effective collaboration, knowledge sharing, and advancements in differentscientific fields.Precision in Engineering1. Design and ManufacturingIn engineering, precision plays a critical role in the design and manufacturing processes. Precise engineering drawings, specifications, and measurements are essential for creating products that meet the desired standards and specifications. Precision in manufacturing ensures that every component fits together perfectly, leading to efficient and reliable systems.2. Quality Control and TestingPrecision is also crucial in quality control and testing within the engineering industry. Precise measurements and inspections are conducted to ensure that products meet specific criteria and standards. Precise testing procedures are essential to identify any defects or deviations from the desired specifications, allowing for necessary adjustments to be made before products are released to the market.3. Safety and ReliabilityPrecision engineering is essential when it comes to ensuring safety and reliability in various engineering applications. For example, in the aerospace industry, precise calculations and meticulous design are necessary to guarantee the integrity of aircraft structures and systems. In the automotive industry, precision in manufacturing processes ensures that vehicles are safe and reliable for consumers.Precision in Mathematics1. Accuracy in CalculationsMathematics is a discipline that heavily relies on precision. From basic arithmetic to complex calculus, precision is crucial to obtain accurate results. A slight deviation in calculations can lead to significant errors, especially when dealing with large-scale computations or solving intricate mathematical problems. Precision in mathematics ensures the reliability and validity of mathematical solutions.2. Rigorous Proofs and TheoremsIn the realm of mathematics, precision is vital in constructing rigorous proofs and theorems. Every step of a mathematical proof must be precise and logical, leaving no room for ambiguity or uncertainty. Precision allows mathematicians to build upon existing knowledge and create new theories and concepts that withstand rigorous scrutiny.3. Mathematical ModelingAnother area where precision is essential in mathematics is mathematical modeling. Mathematical models are used to describe and predict real-world phenomena. By using precise mathematical equations and parameters, mathematicians and scientists can develop models that accurately represent complex systems and events, leading to more reliable predictions and insights.Precision in Language and Communication1. Clarity in Writing and SpeakingPrecision is crucial in language and communication to ensure clarity and understanding. Precise language allows writers and speakers to express their thoughts and ideas with accuracy and specificity. It reduces the chances of misinterpretation or confusion, enabling effective communication between individuals or within a larger audience.2. Technical and Scientific WritingIn technical and scientific writing, precision is of utmost importance. Precise terminology and explanations are necessary to convey complex concepts or procedures accurately. Precision in scientific writing helps researchers and professionals communicate their findings and knowledge effectively, facilitating progress within their respective fields.3. Legal and Contractual MattersPrecision is also crucial in legal and contractual matters. Legal documents, contracts, and agreements need to be precise and unambiguous to avoid misunderstandings or disputes. Precise language ensures thatall parties involved have a clear understanding of their rights, obligations, and responsibilities.ConclusionPrecision plays a vital role in various aspects of our lives, ranging from scientific research and engineering to mathematics and language. It enables accurate measurements, calculations, and communication, leading to advancements in different fields and ensuring reliability, safety, and clarity. Emphasizing precision in our endeavors enhances the quality and effectiveness of our efforts, ultimately contributing to progress and success.。
鲁班造锯的英语,作文Luban's Invention of the SawLuban, also known as Lu Ban, was a renowned ancient Chinese inventor, carpenter, and engineer who lived during the Spring and Autumn period, approximately 500 BCE. He is widely recognized as one of the most influential figures in the history of Chinese technology and is credited with the invention of numerous tools and machines that revolutionized various industries, including construction, woodworking, and even warfare.One of Luban's most significant inventions was the saw, a tool that has become an indispensable part of human civilization. Prior to Luban's innovation, woodworking was a laborious and time-consuming task, often relying on the use of primitive tools such as axes and adzes. These tools, while effective in some applications, were limited in their ability to produce precise and intricate cuts, making the construction of complex structures and detailed woodwork a challenging endeavor.Luban's invention of the saw, however, revolutionized the way people worked with wood. By creating a tool with a serrated blade that could effortlessly slice through wood, Luban opened up new possibilities in the field of woodworking. The saw allowed for more accurate and efficient cutting, enabling carpenters and builders to create intricate designs, complex structures, and a wide range of wooden products with greater ease and precision.The impact of Luban's saw invention cannot be overstated. It not only transformed the construction industry but also paved the way for the development of numerous other tools and machines that relied on the saw as a fundamental component. From the creation of furniture and musical instruments to the construction of ships, bridges, and buildings, the saw became an essential tool in the hands of skilled artisans and craftspeople.Luban's genius, however, extended beyond the invention of the saw. He was also credited with the creation of numerous other tools and machines that left a lasting impact on Chinese civilization. One such invention was the block and tackle, a system of pulleys and ropes that allowed for the efficient lifting and movement of heavy objects. This innovation was particularly valuable in the construction of large-scale structures, as it enabled workers to move and position heavy materials with greater ease and efficiency.Another remarkable invention of Luban's was the drilling machine, a device that revolutionized the way holes were created in wood, stone, and other materials. Prior to Luban's innovation, drilling was a laborious and time-consuming process, often requiring the use of hand-held tools and a significant amount of physical effort. Luban's drilling machine, however, automated the process, allowing for faster and more accurate hole-making, which had far-reaching implications in various industries, from construction to the production of weapons and tools.Luban's contributions to Chinese technology and engineering were not limited to these inventions alone. He is also credited with the development of a variety of military technologies, including the siege ladder, a device used to scale the walls of fortified structures, and the ox-pulled war cart, a mobile platform that could be used to transport soldiers and equipment into battle.The legacy of Luban's inventions and innovations extends far beyond his own lifetime. His creations have continued to shape the course of human history, influencing the development of technology and the way we interact with the world around us. The saw, in particular, has become an indispensable tool in countless industries, and its impact can be seen in the countless products and structures that we rely on in our daily lives.Despite the passage of time, Luban's contributions to Chinese technology and engineering continue to be celebrated and studied by scholars and enthusiasts alike. His innovative spirit and problem-solving abilities have inspired countless generations of inventors and engineers, who have built upon his work to create even more remarkable and transformative technologies.In conclusion, Luban's invention of the saw, as well as his numerous other groundbreaking innovations, stands as a testament to the ingenuity and creativity of the human mind. His legacy continues to inspire and shape the world we live in, and his contributions to the field of technology and engineering will undoubtedly be remembered and celebrated for generations to come.。
看一场台球英语作文Last night, I went to watch a game of pool at the local bar. The atmosphere was lively, with people cheering and clapping as the players made impressive shots. The sound of balls clacking against each other filled the room, creating a sense of excitement and tension.The players were focused and determined, their eyesfixed on the table as they carefully lined up their shots. Each movement was precise and deliberate, with the occasional burst of speed and power as they aimed to sinkthe ball into the pocket.As the game progressed, the tension in the room grew palpable. Spectators held their breath as the players took their shots, their faces a mix of concentration and anticipation. Every successful shot was met with cheers and applause, while missed shots were met with groans and sighs.The players' skill and technique were truly impressive,with each one showcasing their own unique style and strategy. Some favored long, calculated shots, while others preferred quick, aggressive plays. It was fascinating to see how each player approached the game in their own way, adapting to the table and the situation at hand.In the end, there could only be one winner. The final shot was made with a resounding thud, the ball sinking into the pocket with a satisfying clink. The room erupted into cheers and applause as the victorious player raised their cue in triumph, a smile of satisfaction on their face.Overall, it was an exhilarating experience to watch the game of pool unfold before my eyes. The skill, strategy, and intensity of the players made for an unforgettable evening, and I can't wait to go back and watch another game soon.。
网球竞赛英语作文Title: A Thrilling Tennis Competition。
Tennis is not just a sport; it's a blend of strategy, skill, and sheer determination. Recently, I had the exhilarating opportunity to participate in a tennis competition, an experience that left an indelible mark on my sporting journey.The competition was held at a prestigious venue, bustling with excitement and anticipation. As I stepped onto the court, I could feel the energy pulsating through the air. The sun was shining brightly overhead, casting a golden glow on the meticulously manicured courts.My first match was against a formidable opponent, renowned for their powerful serves and agile footwork. It was a nerve-wracking start, with each point fiercely contested. However, I managed to maintain my composure, relying on my training and instinct to guide me through theintense rallies.As the match progressed, I found my rhythm, executing precise shots and anticipating my opponent's moves with greater accuracy. Every stroke felt like a symphony of motion, as I seamlessly transitioned from offense to defense and back again. The crowd was captivated, their applause echoing across the court with each remarkable rally.Despite facing fierce resistance, I refused to back down, drawing upon reserves of determination I never knew I possessed. Every game was a testament to my unwavering resolve, as I pushed myself to the limit in pursuit of victory.In the end, it all came down to the final set, with both players unwilling to concede an inch. The tension was palpable, as each point became increasingly crucial. But in the end, it was my perseverance that prevailed, as I clinched the decisive victory with a well-placed forehand winner.The sense of triumph that washed over me was unparalleled, as I basked in the glow of my hard-earned victory. But more than the thrill of winning, it was the journey itself that left a lasting impression on me. The camaraderie shared with fellow competitors, the adrenaline-fueled battles on the court, and the invaluable lessons learned along the way – all of these are what make tennis such a remarkable sport.In conclusion, my experience in the tennis competition was nothing short of extraordinary. It tested my skills, pushed my limits, and ultimately reaffirmed my passion for the game. And as I look ahead to future competitions, I do so with renewed determination and a deep appreciation for the sport that has given me so much.。
对待猎物英语作文In the wild, the relationship between predator and prey is a fundamental aspect of the ecosystem. The predator's approach to hunting its prey is a complex interplay of instinct, strategy, and survival. Here's an essay that explores this theme:The Art of the Hunt: A Predator's PerspectiveIn the vast expanse of the wilderness, the dance of life and death plays out with an elegance that is both brutal and necessary. The predator, a creature honed by nature to be a master of stealth and cunning, approaches its prey with a single-minded focus. This essay delves into the intricate dynamics of the hunt, as seen through the eyes of the hunter.The Call of InstinctThe predator's instinct is the first to awaken. It is a primal urge, a deep-seated need to feed and survive. This instinct is not born of malice but of the unyielding laws of nature. The predator's senses are heightened; its vision, keen and focused on the slightest movement; its hearing, tuned to the softest rustle of leaves; its sense of smell, acute and able to detect the faintest traces of its prey.The Strategy of the ChaseHunting is not a random act of violence; it is a calculated strategy. The predator must assess its prey, considering factors such as size, speed, and the potential risk of injury. It must also take into account the environment, using the terrain to its advantage. The predator may stalk its prey for hours, patiently waiting for the perfect moment to strike.The Pursuit of SurvivalThe chase is a test of endurance, a battle of wills between predator and prey. The prey, sensing the imminent danger,will use every trick in its arsenal to escape: sudden changes in direction, bursts of speed, and evasive maneuvers. The predator, in turn, must adapt, outmaneuvering its prey with agility and intelligence.The Moment of TruthThe climax of the hunt is the moment of capture. It is aswift and decisive act, the culmination of hours ofpreparation and strategy. The predator's strike must beprecise and efficient, for it is not just about the kill but also about minimizing the risk of injury to itself.The Cycle of LifeAfter the hunt, there is a moment of calm. The predator feeds, fulfilling its need for sustenance. This act is not one of cruelty but of necessity. The predator's role in theecosystem is vital, keeping populations in check and maintaining balance.Reflections on the HuntThe predator's treatment of its prey is a reflection of the natural order. It is a cycle that has been in place for eons, a testament to the resilience and adaptability of life on Earth. As we observe this cycle, we are reminded of the interconnectedness of all living things and the delicate balance that sustains the world.In conclusion, the predator's approach to its prey is a complex narrative of survival, strategy, and instinct. It is a dance as old as time, a reminder of the raw power and beauty of nature's laws. As we learn from the predator, we gain a deeper understanding of our own place within the intricate web of life.。
火药英文作文介绍Title: The Fascinating History and Impact of Gunpowder。
Introduction:Gunpowder, often referred to as black powder, holds a significant place in human history due to itstransformative impact on warfare, technology, and even cultural exchanges. This combustible substance, composed of saltpeter, charcoal, and sulfur, has shaped the course of civilizations and altered the balance of power across the globe.Origins and Development:The origins of gunpowder can be traced back to ancient China, where it was initially developed during the Tang dynasty in the 9th century. Its invention is attributed to Chinese alchemists who sought the elixir of immortality but stumbled upon a mixture with explosive properties instead.Initially utilized for fireworks and later for military purposes, gunpowder gradually spread along trade routes to the Middle East, Europe, and beyond.Impact on Warfare:The introduction of gunpowder revolutionized warfare on a global scale. Prior to its invention, battles relied heavily on traditional weaponry such as swords, spears, and bows. However, the advent of gunpowder weapons such as cannons, muskets, and eventually rifles, transformed the nature of combat. This shift led to the decline of medieval fortifications and the rise of large-scale siege warfare. Empires rose and fell based on their mastery of gunpowder technology, making it a cornerstone of military strategyfor centuries to come.Technological Advancements:The development of gunpowder spurred innovations in metallurgy, chemistry, and engineering. Cannons and firearms required precise craftsmanship and scientificunderstanding to manufacture efficiently. Moreover, theneed for reliable ignition mechanisms led to the inventionof flintlock and percussion cap mechanisms, further advancing firearm technology. These advancements laid the groundwork for modern firearms, paving the way for the industrial revolution and subsequent technological advancements.Cultural Exchanges and Globalization:The spread of gunpowder facilitated cultural exchanges and globalization by connecting distant regions through trade and conquest. European explorers, armed with firearms, ventured across oceans in search of new trade routes and territories. This expansion had profound consequences, leading to the colonization of vast territories and the exchange of goods, ideas, and diseases between continents. Gunpowder not only reshaped military strategies but also influenced art, literature, and societal norms as cultures collided and merged.Ethical and Moral Implications:While gunpowder revolutionized warfare and technological progress, it also raised ethical and moral concerns. The destructive power of firearms and explosives resulted in unprecedented casualties and devastation on the battlefield. Additionally, the proliferation of firearms led to increased violence and conflict in societies worldwide. As a result, debates surrounding gun control, disarmament, and the ethics of warfare persist to this day, highlighting the complex relationship between technology and morality.Conclusion:In conclusion, gunpowder stands as one of the most influential inventions in human history, shaping the course of warfare, technology, and cultural exchange. From its humble origins in ancient China to its global impact on civilization, gunpowder has left an indelible mark on the world. While its legacy is fraught with both triumphs and tragedies, the story of gunpowder serves as a testament tohumanity's ingenuity and its capacity for both creation and destruction.。
全产业链价值创造英文说明书1The concept of full industrial chain value creation refers to the comprehensive and coordinated optimization and integration of all links within an industry chain, from the initial stage of research and development to the final stage of sales and after-sales service. This approach aims to maximize the overall value and competitive advantage of the entire chain.Take a well-known automotive brand as an example. They have achieved value maximization by integrating various aspects such as research and development, production, and sales. In the R&D stage, they invest heavily in technological innovation and design to create unique and appealing vehicle models. During the production process, they adopt advanced manufacturing techniques and strict quality control to ensure high-quality output. In the sales phase, they establish an extensive distribution network and provide excellent customer service to enhance brand image and customer satisfaction.Another case is an electronic enterprise that optimizes its full industrial chain layout to enhance competitiveness. They focus on enhancing the efficiency and flexibility of the supply chain to respond quickly to market changes. They also continuously improve the R&D capabilities to launch new products that meet the diverse needs ofconsumers. At the same time, they build a strong marketing and sales team to expand market share.The significance of full industrial chain value creation is profound. It helps enterprises reduce costs, improve product quality and service levels, and enhance their ability to respond to market fluctuations. Moreover, it promotes the efficient allocation of resources and the upgrading of the entire industry, leading to sustainable development and greater economic benefits.In conclusion, full industrial chain value creation is not only an important strategy for enterprises to succeed in the fierce market competition but also a driving force for the healthy development of the entire industry.2The whole industrial chain value creation is a complex and significant topic that involves multiple elements and challenges. To understand it thoroughly, let's take the example of an agricultural enterprise. In its pursuit of full industrial chain development, it often encounters the risk of market fluctuations. For instance, sudden changes in the demand and supply of agricultural products can lead to price instability. This not only affects the income of farmers but also poses challenges to the processing and sales links. To cope with this, the enterprise needs to establish a precise market monitoring mechanism and a flexible production adjustment strategy.Another example could be a clothing brand. Supply chain issues can have a significant impact on its value creation. Delays in raw material supply or problems in logistics can cause production delays and customer dissatisfaction. To address these problems, the brand should build a stable and efficient supply chain system, strengthen cooperation with suppliers, and improve inventory management.In conclusion, the key elements of whole industrial chain value creation include seamless coordination among various links, effective risk management, and continuous innovation. Only by paying attention to these aspects and taking corresponding measures can enterprises truly achieve sustainable value creation and development in the fierce market competition.3The entire industrial chain value creation represents a revolutionary concept that has reshaped the business landscape in the contemporary era. It involves integrating all stages of production, distribution, and consumption to maximize value and achieve sustainable growth. Take, for instance, a leading internet enterprise that harnessed the power of big data to drive an upgrade across the entire industrial chain. By collecting and analyzing vast amounts of data from various sources, this company was able to identify market trends, customer preferences, and potential operational bottlenecks with unprecedented accuracy. This enabled themto optimize their product offerings, streamline their supply chain, and enhance their marketing strategies, resulting in a significant increase in market share and customer satisfaction.Another compelling example is a traditional manufacturing firm that underwent an intelligent transformation to achieve a breakthrough in value creation. Through the adoption of advanced technologies such as robotics, artificial intelligence, and the Internet of Things, this company automated its production processes, improved product quality, reduced production costs, and shortened delivery times. Simultaneously, it leveraged digital platforms to establish closer connections with customers, providing personalized products and services, and thereby enhancing brand loyalty and competitiveness.In conclusion, the success of the entire industrial chain value creation lies in the seamless integration of resources, the application of innovative technologies, and a customer-centric approach. It requires businesses to have a forward-looking vision, a willingness to embrace change, and the ability to collaborate effectively across different sectors. Only by doing so can enterprises truly unlock the potential of the entire industrial chain and create long-term value in an increasingly competitive marketplace.4The concept of full industrial chain value creation has emerged as a driving force for businesses and society. Let's take a food enterprise as anexample. By implementing full industrial chain management, this enterprise can closely monitor every step from raw material sourcing to production, processing, and distribution. This not only ensures the safety and quality of food but also boosts consumers' trust. For instance, when it comes to the selection of agricultural products, strict standards are imposed to guarantee the freshness and non-pollution of the ingredients. During the production process, advanced technologies and strict quality control measures are adopted to eliminate any potential risks. As a result, consumers are more willing to purchase products from this enterprise, which leads to increased sales and a better reputation.Another example can be found in the energy sector. A certain energy enterprise has made remarkable contributions to promoting the popularization of green energy through the development of a full industrial chain. It starts from the research and development of new energy technologies, followed by the establishment of large-scale production facilities to reduce costs and improve efficiency. Moreover, efforts are made in the construction of energy storage and transmission systems to ensure a stable supply of green energy. This comprehensive approach not only helps reduce reliance on traditional energy sources but also plays a crucial role in protecting the environment and achieving sustainable development.In conclusion, full industrial chain value creation brings numerousbenefits to both enterprises and society. It enhances the competitiveness of enterprises, meets the demands of consumers for high-quality products and services, and contributes to the sustainable development of society as a whole.5The concept of full industrial chain value creation has emerged as a powerful force shaping the dynamics of various industries in today's highly competitive business landscape. It involves the seamless integration and optimization of all stages of a product or service's lifecycle, from raw materials sourcing to end-user consumption.In the financial sector, for instance, the construction of a full industrial chain financial service system has become increasingly crucial. This encompasses providing a comprehensive range of financial products and services, including financing for startups, supply chain finance for enterprises, and wealth management for individuals. By integrating these elements, financial institutions can better meet the diverse needs of clients and enhance their overall competitiveness.The healthcare industry has also witnessed significant improvements through full industrial chain integration. By integrating various components such as medical research and development, production of medical devices and drugs, hospital operations, and post-treatment rehabilitation, the allocation of medical resources can be optimized. Thisresults in improved accessibility and quality of healthcare services for patients.Looking forward, the trend of full industrial chain value creation is set to continue and intensify. Industries will need to focus on technological innovation, data analytics, and strategic partnerships to further enhance the efficiency and effectiveness of their value creation processes. Only by embracing this holistic approach can businesses thrive and contribute to sustainable economic growth and social development.。
人民日报农民增收模式英文翻译在我国,农民增收一直是一个备受关注的话题。
人民日报对于农民增收模式的英文翻译提供了一种新的视角和思路。
在本文中,我们将从多个维度探讨人民日报对农民增收模式的英文翻译,深入剖析其内涵和实践意义。
1. What is "人民日报农民增收模式"?人民日报农民增收模式指的是一种旨在帮助农民提高收入的体系或方法。
这种模式涵盖了诸多领域,包括农业技术、市场开拓、产销对接、金融扶持等。
它的出现旨在解决农民收入增长缓慢、发展不平衡的问题,促进农村经济的健康发展。
2. The translation of "人民日报农民增收模式"The translation of "人民日报农民增收模式" can be "Peoples' Daily Farmers' Income Increase Model". This translation accurately conveys the original meaning of the term and maintains the coherence and conciseness of the language. Through this translation, the essence of the model is effectively presented to the international community.3. The significance of "人民日报农民增收模式"The significance of "人民日报农民增收模式" lies in its potential topromote rural revitalization and achieve common prosperity. By improving farmers' income, it not only enhances their living standards but also stimulates rural consumption and boosts the overall economic growth. It represents a fundamental strategy for building a moderately prosperous society in all respects.4. Personal insightsFrom my perspective, "人民日报农民增收模式" embodies the Chinese government's commitment to inclusive development and poverty alleviation. It reflects the holistic approach of targeted poverty reduction and the implementation of precise measures to benefit the rural population. As an essential part of China's rural revitalization strategy, it demonstrates the government's determination to narrow the urban-rural income gap and promote balanced development.In conclusion, the translation of "人民日报农民增收模式" as "Peoples' Daily Farmers' Income Increase Model" has accurately conveyed the original meaning and significance of the concept. This model represents a comprehensive and innovative approach to rural development and poverty alleviation, and its successful implementation will contribute to the realization of common prosperity in China.以上就是我根据你提供的主题撰写的一篇文章,希望能为你提供帮助。
模具设计与制造外文翻译参考文献(文档含中英文对照即英文原文和中文翻译)译文:模具设计与制造模具是制造业的重要工艺基础,在我国模具制造属于专用设备制造业。
中国虽然很早就开始制造模具和使用模具,但长期未形成产业。
直到20世纪80年代后期,中国模具工业才驶入发展的快车道。
近年,不仅国有模具企业有了很大发展,三资企业、乡镇(个体)模具企业的发展也相当迅速。
虽然中国模具工业发展迅速,但与需求相比,显然供不应求,其主要缺口集中于精密、大型、复杂、长寿命模具领域。
由于在模具精度、寿命、制造周期及生产能力等方面,中国与国际平均水平和发达国家仍有较大差距,因此,每年需要大量进口模具。
中国模具产业除了要继续提高生产能力,今后更要着重于行业内部结构的调整和技术发展水平的提高。
结构调整方面,主要是企业结构向专业化调整,产品结构向着中高档模具发展,向进出口结构的改进,中高档汽车覆盖件模具成形分析及结构改进、多功能复合模具和复合加工及激光技术在模具设计制造上的应用、高速切削、超精加工及抛光技术、信息化方向发展。
近年,模具行业结构调整和体制改革步伐加大,主要表现在,大型、精密、复杂、长寿命、中高档模具及模具标准件发展速度高于一般模具产品;塑料模和压铸模比例增大;专业模具厂数量及其生产能力增加;“三资”及私营企业发展迅速;股份制改造步伐加快等。
从地区分布来看,以珠江三角洲和长江三角洲为中心的东南沿海地区发展快于中西部地区,南方的发展快于北方。
目前发展最快、模具生产最为集中的省份是广东和浙江,江苏、上海、安徽和山东等地近几年也有较大发展。
虽然我国模具总量目前已达到相当规模,模具水平也有很大提高,但设计制造水平总体上落后于德、美、日、法、意等工业发达国家许多。
当前存在的问题和差距主要表现在以下几方面:(1)总量供不应求,国内模具自配率只有70%左右。
其中低档模具供过于求,中高档模具自配率只有50%左右。
(2)企业组织结构、产品结构、技术结构和进出口结构均不合理。
Targeted Poverty Relief China’s New Anti-Poverty StrategySince the late 1970s,China has made great progress in large-scale poverty reduction in the process of its reform and opening up,making considerable contribution to the realization of the UN Millennium Development Goals(MDGs). However,the nation is still facing problems such as income inequality,comparatively lower poverty line,huge poor population,and lack of targeted poverty alleviation measures. In this context,the strategy of “targeted poverty relief”needs to be taken further for hastening large-scale poverty reduction. Progress in Poverty AlleviationOver nearly four decades since it began the reform and openingup policy,China has greatly reduced poverty while maintaining rapid economic growth. According to the World Bank’s poverty line of US$1 a day,the country cut the number of the poor by more than 700 million in the past 40 years. In 2000,the UN Millennium Summit passed the MDGs,setting a goal to halve the number of the poor from the 1990 figure. China was the first country to reach the goal. By 2002,it had reduced the percentage of the poor in rural areas to 30percent,which was 60 percent in 1990. During the period,China’s contribution rate to global poverty reduction exceeded 70 percent. China has made further efforts to reduce poverty since the 18th National Congress of the Communist Party of China in 2012. By the end of 2017,the population of the poor in the country’s rural areas had decreased to 30.26 million from 98.99 million at the end of 2012,and the poverty headcount ratio had dropped from 10.2 percent to 3.1 percent.Rapid economic growth over decades has substantially increased incomes and consumption,being decisive inlarge-scale poverty reduction. This large-scale poverty reduction is attributed to a combination of many factors. Firstly,growth in agriculture is particularly important for poverty alleviation. According to a research report released by the World Bank,growth in China’s agricultural sector is four times as effective in reducing poverty as growth in the secondary and tertiary industries. Secondly,targeted poverty alleviation effort has helped narrow the gap between different regions and accelerate economic and social development ofpoverty-stricken areas. Thirdly,China began to establish a social security system for rural areas in 2007. Measures such as guaranteed minimum income,the new-type rural cooperativemedical care system and the rural pension insurance system have ensured provision of basic living and public services. Fourthly,inclusive rural policies have benefited vast sections of the impoverished in rural areas. In 2003,the country launched the “grain for green”campaign in poverty-stricken areas,and farmers who returned their farmland to forests and grasslands were paid with allowances. In 2006,agricultural tax was abolished,and a policy to provide general subsidies for agricultural development was implemented. From 2008,nine-year free compulsory education began to be available nationwide for all children. And finally,the basic land system and land operation pattern ensure that the poor benefit from agricultural growth. The household contract responsibility system was adopted as the basic land system in rural areas,according to which rural lands are collectively owned,but farmers enjoy long-term use and management rights of the land contracted. In the early 1980s,farmlands were distributed to farmers in a basically equal manner,so that impoverished households could also benefit from their farmland and agricultural development.Challenges in Poverty ReductionDespite great progress in economic development andpoverty reduction,it is undeniable that the income gap between the rich and the poor continues to expand. China’s Gini coefficient grew from 0.288 in 1981 to 0.4 in 2017. As relief measures accelerated economic growth of poverty-stricken regions,income inequality in those regions continued to increase. During the implementation of the Outline for Development-oriented Poverty Reduction for China’s Rural Areas(2001-2010),the ratio of the average income of the poorest households to that of the richest households in key counties for poverty alleviation dropped from 21.59 percent in 2002 to 17.38 percent in 2010. The more income a rural household earns,the faster its net income grows. From 2002 to 2010,the per-head net income of farmers with the lowest incomes increased at an annual rate of 11.1 percent,while that of farmers with the highest incomes increased at an annual rate of 14.1 percent,resulting in a further expansion of the income gap. From 2002 to 2009,the annual income growth rates for poor rural households and average rural households in key counties for poverty alleviation were 2.75 percent and 11.76 percent,respectively―the former is 9 percent lower than the latter,while the national average stayed at 11.04 percent. The ratio of the income of poor households to that ofthe average households nationwide continued to fall―from one third in 2002 to one fifth in 2009.The increase in income inequality is attributed to various reasons. First,the mainstay of China’s economic structure has shifted from agriculture that is labor-intensive to manufacturing and service industries,resulting in further expansion of the income gap. Due to the fact that the country adopts a land system featuring equal distribution,the income distribution in its agricultural sector is comparatively equal. However,income inequality grows in the secondary and tertiary industries that are capital- and technologyintensive. Second,the population and labor migration between rural areas and urban areas also causes income inequality. Due to their comparatively lower educational level and comprehensive capacity and the shortage of capital and information,migrant workers from povertystricken rural areas are less likely to find jobs in cities than those from comparatively richer rural areas. This further widens the income gap. Third,although development-oriented relief effort has greatly improved infrastructure,production,and living conditions in poverty-stricken areas,it remains hard for poor households to substantially increase their incomes by utilizing improved infrastructure as rich households did. Finally,it is difficult for the poor to enjoy effective financial services,which impedes a rise in their incomes. Against the backdrop of growing income inequality,it has become harder to reduce the number of the poor through economic development and regional poverty alleviation. Therefore,it is increasingly important to directly help the poor through more targeted relief policies.Targeted Poverty Alleviation StrategyIn November 2013,Chinese President Xi Jinping first put forward the strategy of “targeted poverty alleviation”during his visit to Xiangxi Tujia and Miao Autonomous Prefecture in Hunan Province. So far,the strategy has remained a significant part of China’s fight against poverty.The strategy aims to enhance the relevance and efficacy of relief effort,so as to offset the drop in the effect of economic growth on poverty reduction. The key content of targeted poverty alleviation is elimination of all the factors and obstacles that cause poverty through targeted assistance for the poor and enabling their selfdevelopment towards the goal of sustainable poverty reduction. Targeted poverty alleviation includes precise identification,assistance,management,and assessment.According to a survey conducted by the National Bureau ofStatistics of China,the country had 70.17 million below the poverty line at the end of 2014. (It was estimated that the figure would be reduced to 60 million in early 2016). The government has taken a series of measures to further innovate its poverty relief mechanism,so as to facilitate the implementation of its targeted poverty alleviation strategy and ensure eradication of poverty by 2020. In terms of precise identification,China has identified 29.48 million poor households with a total population of 89.62 million since 2013. (Currently,the second round of poor identification is underway,and although the findings are yet to be revealed,it is believed that the identified poor would be less in numbers). Moreover,the country has registered all poor households and population and details about the families,as well as their available resources,income sources,and reasons of poverty,in the national poverty alleviation information system.The government has taken a series of measures to push forward targeted poverty alleviation. First,supporting a batch of poor households through industrial development and employment and solving their difficulties in relation to technology,capital,and marketing. Second,relocating 10 million of the poor in remote areas with harsh naturalconditions to comparatively more hospitable villages or small towns with a view to improving the environment and conditions for their development. Third,helping a batch of poor households reduce poverty through ecological compensation policies such as subsidies for those who returned their farmland to forests. Fourth,helping a batch of poor households reduce poverty through strengthening education. The measures include developing preschool education in povertystricken areas,providing free high school or occupational school education and living allowances for students from impoverished families. This will not only reduce the education expenditure of poor households,but also help end inter- generational poverty. Finally,helping a batch of poor households through social security measures such as expanding the coverage of minimum living allowances,launching rural cooperative medical care system,and providing severe-disease medical insurance and assistance,and pension insurance. By 2020,China’s minimum living security system will lift all its citizens above the poverty line. To strengthen communitylevel poverty alleviation capacity,governments at various levels have dispatched officials to act as first secretaries and poverty relief team leaders in 128,000poverty-stricken villages. Academic institutions,nongovernmental organizations(NGOs),and ordinary citizens are also encouraged to participate in poverty relief efforts by various means,such as establishing independent third-party poverty reduction appraisal mechanism.Chinese Experience in Poverty ReliefChina’s success in large-scale poverty reduction over the past four decades,as well as its ongoing effort in targeted poverty alleviation,is useful experience for developing countries. China’s successful experience can be summarized as follows:First,sustained economic growth has been combined with development-centered relief effort. For any country,economic growth is a necessary prerequisite for largescale poverty reduction. Against a backdrop of increasing income inequality,targeted poverty relief is indispensable to end poverty.Second,the country has realized an organic integration of poverty alleviation and social security. Essentially,in order to achieve sustainable poverty reduction,a country needs to enhance the selfdevelopment ability of povertystricken areas and population through development-oriented relief effort.Moreover,providing necessary social security policies will not only guarantee the poor’s basic livelihood,but also lay the foundation for development-oriented poverty alleviation. If the poor lack basic living guarantee,it is impossible to achieve sustainable poverty reduction.Third,government-led relief effort should be made alongside social mobilization. With primary liability for poverty alleviation,governments at various levels are responsible for formulating relief strategies,providing and mobilizing relevant resources,and drafting and implementing relevant plans and policies. Poverty alleviation is a comprehensive,systemic project that involves various sectors and requires specialized knowledge and expertise. Therefore,broad participation of market entities,NGOs and citizens is vital to enhancing the effectiveness of poverty relief efforts.Finally,rational institutional arrangements are helpful for poverty reduction. China’s fair land distribution system has considerably magnified agriculture’s role in poverty reduction. Moreover,the popularization of free compulsory education and cooperative medical care helps the poor increase their human capital and enhance their capacity for development.。
IntroductionThe pharmaceutical industry is an intricate and highly specialized sector that relies heavily on precise communication, particularly in terms of technical terminology. To ensure the development, production, and distribution of high-quality medications, professionals within this field must be well-versed in a vast array of English terms that encompass various aspects of drug discovery, clinical trials, regulatory affairs, manufacturing, marketing, and pharmacovigilance. This comprehensive analysis aims to delve into these essential terms, providing a multifaceted and in-depth understanding of their significance in the context of the pharmaceutical industry.1. Drug Discovery and Developmenta) **Lead compound**: The initial molecule identified during the drug discovery process with promising biological activity against a specific therapeutic target. It serves as the starting point for further optimization.b) **Target identification and validation**: The process of identifying a specific molecular entity (e.g., protein, gene) involved in a disease pathway and demonstrating its potential as a therapeutic intervention.c) **High-throughput screening (HTS)**: A rapid, automated method used to screen large libraries of chemical compounds or biological molecules for their ability to modulate a specific biological target or pathway.d) **Structure-activity relationship (SAR)**: The study of how changes in the chemical structure of a molecule affect its biological activity, providing insights for lead optimization.e) **Pharmacokinetics (PK)**: The study of how a drug is absorbed, distributed, metabolized, and excreted by the body, determining its bioavailability, half-life, and exposure.f) **Pharmacodynamics (PD)**: The study of the biochemical and physiological effects of a drug on the body, including its mechanism of action, potency, and efficacy.g) **Preclinical studies**: Non-clinical research conducted in vitro (e.g., cell cultures) and in vivo (e.g., animal models) to assess the safety, efficacy, and PK/PD properties of a drug candidate before human trials.2. Clinical Trialsa) **Investigational New Drug (IND) application**: A regulatory filing submitted to a health authority (e.g., FDA, EMA) seeking permission to conduct human clinical trials with an investigational drug.b) **Phase I, II, III, and IV clinical trials**: Sequential stages of human testing designed to evaluate the safety, tolerability, dosing, efficacy, and long-term safety of a drug candidate, respectively.c) **Randomized controlled trial (RCT)**: A rigorous study design where participants are randomly assigned to receive either the experimental treatment or a control (placebo or standard therapy), minimizing bias in assessing treatment effects.d) **Adverse event (AE)**: Any untoward medical occurrence experienced bya participant during a clinical trial, whether or not it is considered related to the investigational drug.e) **Serious adverse event (SAE)**: An AE that results in death, is life-threatening, requires hospitalization, or causes significant disability or incapacity.f) **Endpoint**: A predefined measurement or outcome used to assess the effectiveness of a treatment in a clinical trial, such as reduction in symptoms, improvement in survival rate, or change in biomarker levels.3. Regulatory Affairsa) **New Drug Application (NDA)**: A comprehensive submission to a regulatory agency containing data from preclinical and clinical studies, manufacturing information, and proposed labeling, seeking marketing approval for a new pharmaceutical product.b) **Abbreviated New Drug Application (ANDA)**: A regulatory filing for generic drugs, demonstrating bioequivalence to a reference-listed drug without repeating the full spectrum of preclinical and clinical studies.c) **Good Laboratory Practice (GLP)**, **Good Clinical Practice (GCP)**, **Good Manufacturing Practice (GMP)**: Sets of quality standards ensuring the reliability, integrity, and ethical conduct of non-clinical studies, clinical trials, and pharmaceutical production, respectively.d) **Marketing Authorization (MA)**: Official approval granted by a regulatory agency to market and distribute a pharmaceutical product within a specific jurisdiction.e) **Post-marketing surveillance (PMS)**: Ongoing monitoring of a marketed drug's safety profile, effectiveness, and risk-benefit balance in real-world settings.4. Pharmaceutical Manufacturinga) **Active pharmaceutical ingredient (API)**: The biologically active component of a drug product responsible for its therapeutic effect.b) **Excipient**: An inactive substance included in a drug formulation to aid in manufacturing, stability, bioavailability, taste, or other desired properties.c) **Formulation development**: The process of designing and optimizing the composition of a drug product, including API, excipients, and dosage form (e.g., tablet, capsule, injection).d) **Process validation**: Establishing documented evidence that a manufacturing process consistently produces a product meeting predetermined quality specifications and performance characteristics.e) **Batch release**: The authorization for a specific batch of a drug product to be released for distribution after satisfactory completion of all required quality control tests and documentation.5. Pharmaceutical Marketing and Salesa) **Branding**: The strategic process of creating and promoting a unique identity and image for a pharmaceutical product to differentiate it fromcompetitors and enhance its market position.b) **Product lifecycle management (PLM)**: A holistic approach to managing the various stages of a drug's commercial existence, from launch to patent expiration or withdrawal, encompassing pricing, promotion, and market adaptation strategies.c) **Market access**: The ability of patients to obtain and afford a pharmaceutical product, influenced by factors such as reimbursement policies, pricing negotiations, and distribution channels.d) **Key opinion leader (KOL)**: A respected healthcare professional with expertise in a specific therapeutic area who influences the opinions and prescribing behaviors of their peers and can serve as a valuable collaborator for pharmaceutical companies.6. Pharmacovigilancea) **Adverse drug reaction (ADR)**: A response to a medicinal product that is noxious and unintended, occurring at doses normally used in humans for prophylaxis, diagnosis, or therapy.b) **Spontaneous reporting system**: A system for collecting reports of suspected ADRs from healthcare professionals, patients, and others on a voluntary basis.c) **Signal detection**: The process of identifying potential new, unknown, or changing risks associated with a medicinal product based on pharmacovigilance data.d) **Risk management plan (RMP)**: A document outlining the strategy for identification, assessment, mitigation, and communication of risks associated with a medicinal product throughout its lifecycle.ConclusionThe pharmaceutical industry is characterized by a rich lexicon of specialized English terminology, reflecting the complexity and interdisciplinary nature of the work carried out in this sector. From drug discovery and development to clinical trials, regulatory affairs, manufacturing, marketing, and pharmacovigilance, each aspect is underpinned by a distinct set of terms that facilitate precise communication and ensure adherence to rigorous scientific, ethical, and quality standards. Mastery of this terminology is crucial for professionals working in the pharmaceutical industry to effectively contribute to the development and delivery of high-quality, safe, and efficacious medications for patients worldwide.。
A Strategy for Precise and Large ScaleIdentification of Core FucosylatedGlycoproteins*□SWei Jia‡§¶,Zhuang Lu‡¶ʈ,Yan Fu¶**,Hai-Peng Wang**,Le-Heng Wang**,Hao Chi**,Zuo-Fei Yuan**,Zhao-Bin Zheng‡,Li-Na Song‡,Huan-HuanHan‡,Yi-Min Liang‡,Jing-Lan Wang‡,Yun Cai‡,Yu-Kui Zhang ʈ,Yu-Lin Deng ʈ,Wan-Tao Ying‡‡‡,Si-Min He**§§,and Xiao-Hong Qian‡¶¶Core fucosylation (CF)patterns of some glycoproteins are more sensitive and specific than evaluation of their total respective protein levels for diagnosis of many diseases,such as cancers.Global profiling and quantitative charac-terization of CF glycoproteins may reveal potent biomar-kers for clinical applications.However,current tech-niques are unable to reveal CF glycoproteins precisely on a large scale.Here we developed a robust strategy that integrates molecular weight cutoff,neutral loss-depend-ent MS 3,database-independent candidate spectrum fil-tering,and optimization to effectively identify CF glyco-proteins.The rationale for spectrum treatment was innovatively based on computation of the mass distribu-tion in spectra of CF glycopeptides.The efficacy of this strategy was demonstrated by implementation for plasma from healthy subjects and subjects with hepatocellularcarcinoma.Over 100CF glycoproteins and CF sites were identified,and over 10,000mass spectra of CF glycopep-tide were found.The scale of identification results indi-cates great progress for finding biomarkers with a partic-ular and attractive prospect,and the candidate spectra will be a useful resource for the improvement of database searching methods for glycopeptides.Molecular &Cel-lular Proteomics 8:913–923,2009.Glycoproteins are implicated in a wide range of biological processes such as fertilization,development,the immune response,cell signaling,and apoptosis.Altered glycosylation patterns can affect the conformations of glycoproteins and their functions and interactions with other molecules (1,2).Abnormal glycosylation has been demonstrated in many pathological processes.Targeted glycosylation research is considered increasingly important as a way to find novel therapeutic approaches (2,3),and core fucosylation (CF)1glycoproteomics has attracted particularly great attention (4,5).Previous reports show that CF glycoproteins are involved in many important physiological processes,such as trans-forming growth factor-1(6)and epidermal growth factor signaling pathways (7).They also play key roles in many pathological processes,such as hepatocellular carcinoma (HCC)(8,9),pancreatic cancer (10,11),lung cancer (6,12),ovarian cancer (13),and prostate cancer (14).Moreover the CF patterns of several glycoproteins have been reported to serve as more sensitive and specific biomarkers than their total respective protein levels (8,9,15,16).The combination of a biomarker panel of CF glycoproteins is expected to serve as a more reliable diagnostic standard (13).Glycoproteomics research has been conducted for several years and has led to the generation of many effective evalu-ation methods.Most of these methods use lectin or the chem-ical reagent hydrazide to enrich glycopeptides.The oligosac-charide chains are then completely released by treatment ofthe glycopeptides with peptide-N -glycosidase F.Finally thedeglycosylated peptides and the deglycosylation sites areidentified by tandem mass spectrometric analysis (17,18).Although impressive results have been attained,this com-monly used strategy is not an ideal choice for CF glycopro-teins research.First,the enrichment specificity of lectin is not satisfactory (19)as hydrazide chemical reactions irreversiblydestroy glycan structures,particularly fucose tags.Second,the deglycosylation site is determined by the 0.9840-Da mass shift caused by the asparagine to aspartic acid transfer;its confidence can be compromised by deamination of the Asn.Besides that,the CF site can no longer be distinguished fromother glycosylation sites in the same glycoprotein.Thus,the ideal way to precisely identify CF glycoproteins on a large From the ‡State Key Laboratory of Proteomics-Beijing Proteome Research Center-Beijing Institute of Radiation Medicine,No.33Life Science Park Road,Changping District,Beijing 102206,China,§In-stitute of Biophysics,Chinese Academy of Sciences,No.15DatunRoad,Chaoyang District,Beijing 100101,China,ʈBeijing Institute of Technology,No.5South Zhongguancun Street,Haidian District,Bei-jing 100081,China,and **Institute of Computing Technology,Chi-nese Academy of Sciences,No.6Kexueyuan South Road,Beijing 100190,China Received,November 5,2008,and in revised form,January 7,2009Published,MCP Papers in Press,January 12,2009,DOI 10.1074/mcp.M800504-MCP2001The abbreviations used are:CF,core fucosylation;HCC,hepato-cellular carcinoma;rhEPO,recombinant human erythropoietin;RP,reversed phase;S 2,symbol ion 2;S 3,symbol ion 3;HS,Hereman-Schmid;SCX,strong cation exchange;LTQ,linear trap quadrupole.Research©2009by The American Society for Biochemistry and Molecular Biology,Inc.Molecular &Cellular Proteomics 8.5913This paper is available on line at by Xiaohong Qian on May 1, 2009 Downloaded from /DC1/cgi/content/full/M800504-MCP200Supplemental Material can be found at:scale is to provide direct evidence for the existenceof CF modification.Traditional approaches,such as lectin blots,are not sufficiently powerful to meet this requirement.Instead recent advancements in high end MS-based techniques have ignited the hope to reach this challenging goal (20,21).Our group has developed an innovative and systematic strategy for the precise and large scale identification of CF glycoproteins.Several steps were taken leading up to the development of our strategy.1)We established a novel en-richment step for CF glycopeptides,combining the use of lectin for CF glycoprotein enrichment with ultrafiltration for further enrichment of glycopeptide.Glycopeptide enrichment by ultrafiltration based on molecular weight cutoff technology has the added merit of integrating enrichment,desalting,and concentration into a one-step operation.2)We established a neutral loss-dependent MS 3scan method that specificallycaptures partially deglycosylated CF glycopeptides (with fu-cosyl-N -acetylglucosamines residue retained).In MS 3,the intensity distribution of the fragment peaks is much more homogeneous,and there are fewer theoretical fragment ions and interfering peaks than in MS 2.3)We established a novel database-independent candidate spectrum-filtering method for selecting partially deglycosylated CF glycopeptides and a spectrum optimization method.By introducing several strict and appropriate criteria into a scoring system,high quality candidate spectra can be selected before searching the da-tabase,which not only increases the database search effi-ciency but also improves the identification credibility.Further-more by statistically analyzing candidate spectra,some important glycan-related fragmentation patterns were re-vealed.Based on these observations,many kinds of interfer-ing peaks due to glycan fragmentation that are always very intensive and would decrease the accuracy of peptide scoring can be localized and removed from the spectra.This treat-ment can effectively increase the number of identifications through database searching or de novo analysis.The efficacy of this strategy was testified by implementing it on both healthy and HCC plasma.Respectively,105and 106CF sites were identified from 72and 79glycoproteins,includ-ing 19annotated potential glycosylation sites and 25novel ones.This study holds promise for the large scale determina-tion of core fucosylated biomarker panels from clinical sam-ples,either body fluids or tissue biopsies.EXPERIMENTAL PROCEDURESMaterials—The apotransferrin,fetuin,ribonuclease B,endoglyco-sidase F3,formic acid,TFA,␣-cyano-4-hydroxycinnamic acid,and Lens culinaris lectin (agarose conjugate,saline suspension)were pur-chased from Sigma,methyl-␣-D -mannopyranoside was purchased from Fluka (St.Louis,MO),and sodium-3-[(2-methyl-2-undecyl-1,3-dioxolan-4-yl)methoxy]-1-propanesulfonate (RapiGest TM SF)waspurchased from Waters.Sequencing grade porcine trypsin was pur-chased from Promega (Madison,WI);IgG was purified by use of a HiTrap Protein G HP column from GE Healthcare.The PD-10desalt-ing column was also from GE Healthcare.Deionized water was pro-duced by a Milli-Q A10system from Millipore (Bedford,MA).HPLC-grade quality ACN was purchased from J.T.Baker Inc.Iodoacetamide and DTT were obtained from ACROS.The Handee F IG .1.The efficiency of the ultrafil-tration method for enriching glyco-peptide.MS spectra from ultrafiltrationexperiments are shown with the reten-tate fraction (top ),filtrate fraction (mid-dle ),and untreated fraction (bottom ).Glycopeptide C #GLVPVLAENYN*K (A )from apotransferrin only appeared in theretentate fraction.LC #PDC #PLLA-PLN*DSR (B ),VVHAVEVALATFNAESN*-GSYLQLVEISR (F ),and RPTGEVYDIEID-TLETTC #HVLDPTPLAN*C #SVR (G )werefrom fetuin;GQALLVN*SSQPWEPLQ-LHVDK (C )and EAEN*ITTGC #AEHC-#SLNEN*ITVPDTK (E )were from rhEPO;QQQHLFGSN*VTDC #SGNFC #LFR (D )was from apotransferrin.*,annotated gly-cosite;#,carbamidomethylation.Precise and Large Scale CF Glycoprotein Identification914Molecular &Cellular Proteomics 8.5 by Xiaohong Qian on May 1, 2009 Downloaded frommini spin column kit was purchased from Pierce.TheC 18ZipTip and Microcon YM-3were purchased from Millipore.Recombinant human erythropoietin (rhEPO)was a gift from the National Institute for the Control of Pharmaceutical and Biological Products.Healthy human plasma (0.8ml for each experiment)was obtained from a healthy donor.Samples of hepatocellular carcinoma plasma were mixed from eight patients with 0.1ml from each person.IgG Extraction—Plasma was supplemented with IgG binding buffer (20m M sodium phosphate,pH 7.0),and then IgG was depleted by trapping on a column of HiTrap Protein G.The unbound samples were desalted by a PD-10column.Lectin Affinity—Samples were supplemented with 1.6ml of lectin binding buffer (20m M Tris-buffered saline,0.3M NaCl,1m M MnCl 2,1m M CaCl 2,pH 7.4).The samples were incubated for 16h at 4°C with L.culinaris lectin in a spin column (about 300l of lectin-agarose and 400l of sample in each column).After unbound proteins were re-moved by washes with binding buffer,the CF glycoproteins were eluted with elution buffer (binding buffer supplemented with 200m M ␣-D -methylmannoside),then desalted (by PD-10column),and lyophilized.Reduction,Alkylation,and Trypsin Digestion—Samples were dis-solved in 200l of solution that contained 8M urea and 5m M DTT and were reduced at 37°C for 4h.Then iodoacetamide was added to the solution (final concentration,15m M ),which was then further incu-bated for 1h in darkness at room temperature.Afterward 50m M NH 4HCO 3was added to reduce the concentration of urea below 1M ,and sequencing grade trypsin was added at a ratio of enzyme to protein of 1:50.The mixture was then vortexed and incubated at 37°C overnight.0.1%RapiGest SF was used instead of urea for protein denaturation in the repeat experiment of healthy and HCC plasma.TFA was added to the digested protein samples (final TFA concentration was 0.5%,pH Ͻ2),and the samples were incubated at 37°C for 45min.Finally the acid-treated samples were centrifuged at 13,000rpm for 10min,and the supernatants were collected.Enrichment,Desalting,and Concentration of Glycopeptides—Tryp-tic digests were pipetted into Microcon YM-3centrifugal filter de-vices.The absolute amount of glycoprotein in the digests was between 200and 300g for each filter device,and the sample volume was diluted to 500l for each filter device.The samples were centrifuged at 8000ϫg to reduce the sample volume from 500l to about 20l;this required about 3h.Then 450l of deionized water were added to the reservoir and centrifuged at 8000ϫg for 3h;this was repeated twice.After that,the retentate fraction was transferred to a vial,and the reservoir was thrice washed with 20%ACN.All of the retentate fractions and wash solutions were pooled and lyophilized.Endoglycosidase F3Digestion—Glycopeptides were resuspended in 100l of sodium acetate solution (50m M ,pH 4.5)and then incubated with endoglycosidase F3overnight at 37°C.Ammonium acetate (50m M ,pH 4.5)was used instead of the sodium acetate in the repeat experiments of healthy and HCC plasma.Strong Cation Exchange (SCX)Peptide Fractionation—10%en-riched samples were directly analyzed with RP HPLC-MS two times.Other enriched CF glycopeptides were reconstituted with 300l of 5m M ammonium chloride,pH 3.0,25%acetonitrile and fractionated by SCX chromatography on a BioBasic SCX 250ϫ4.6-mm column (Thermo Fisher).The particle size of the column was 5m and pore size was 300Å.The separations were performed at a flow rate of 0.5ml/min using the Elite HPLC system,and mobile phases consisted of 5m M ammonium chloride,pH 3.0,25%acetonitrile (A)and 500m M ammonium chloride,pH 3.0,25%acetonitrile (B).After loading 300l of sample onto the column,the gradient was maintained at 100%A for 10min.Peptides were then separated using a gradient of 0–15%B over 1min followed by a gradient of 15–50%B over 49min.Then the gradient was changed to 50–100%over 5min.The gradient was then held at 100%B for 5min.A total of 15fractions were collected,and each fraction was dried under vacuum.F IG .2.The neutral loss peaks in MS 2spectra of partially deglycosylated CFglycopeptides.The intensities of thehighest peaks are several times higherthan that of the second most intensepeak in all of these MS 2spectra in theion trap,resulting from loss of the fucoseresidue in CID.a ,b ,and c are MS 2spec-tra from the same partially deglycosy-lated CF glycopeptide,EEQYJSTYR(from human IgG).Intensities of the basepeaks were 1.86e 5,2.10e 4,and 2.53e 3,respectively.d and e are MS 2spectra ofsimplified CF glycopeptides GQA-LLVJSSQPWEPLQLHVDK (intensity,3.21e 4;from rhEPO)and QQQHLFG-SJVTDC #SGNFC #LFR (intensity,7.59e 4;from apotransferrin).The MS 2spectra inFT-ICR were collected to check theidentities of the strongest peaks:f forIgG,g for d ,and h for e .J,CF site;#,carbamidomethylation.Precise and Large Scale CF Glycoprotein IdentificationMolecular &Cellular Proteomics 8.5915 by Xiaohong Qian on May 1, 2009 Downloaded fromRP HPLC-MS n Analysis—RP HPLC-MS n experiments wereper-formed on an LTQ-FT mass spectrometer (Thermo Fisher)equipped with a nanospray source and Agilent 1100high performance liquid chromatography system (Agilent Technologies).Peptide mixes were separated on a fused silica microcapillary column with an internal diameter of 75m and an in-house prepared needle tip with an internal diameter of ϳ15m.Columns were packed to a length of 10cm with a C 18reversed phase resin (GEAgel C 18SP-300-ODS-AP;particle size,5m;pore size,300Å;Jinouya,Beijing,China).Sepa-ration was achieved using a mobile phase from 1.95%ACN,97.95%H 2O,0.1%FA (phase A)and 79.95%ACN,19.95%H 2O,0.1%FA (phase B),and the linear gradient was from 5to 50%buffer B for 80min at a flow rate of 300nl/min.The LTQ-FT mass spectrometer was operated in the data-dependent mode.A full-scan survey MS exper-iment (m /z range from 400to 2000;automatic gain control target,5e 5ions;resolution at 400m /z ,100,000;maximum ion accumulation time,750ms)was acquired by the FT-ICR mass spectrometer,and the five most abundant ions detected in the full scan were analyzed by MS 2scan events (automatic gain control target,1e 4ions;maximum ion accumulation time,200ms).The scan model of MS 2was set as the profile.An MS 3spectrum was automatically collected when one of the three most intense peaks from the MS 2spectrum corre-sponded to a neutral loss event of 73.0290m /z ,48.6860m /z ,or 36.5145m /z (charges of parent ions were not collected).The normal-ized collision energy was 35.On-line Two-dimensional LC-MS n —The autosampler was used to inject samples onto the SCX column (BioX-SCX,5cm)after which they were eluted onto a trap column using a stepwise gradient of 0,20,30,40,50,60,70,80,90,and 100%SCX-B.Peptides on the trap column were desalted and then eluted onto the RP column and into the mass spectrometer (the same method as RP HPLC-MS n analysis,but the linear gradient was from 5to 50%buffer B for 120min).Mobile phase buffer for SCX-A was 10m M citric ammonia buffer,pH 3.0,and mobile phase buffer for SCX-B was 50m M citric ammonia buffer,pH 8.5.Experiments of HCC samples were analyzed by this system (Eksigent NanoLC-2D)and repeated one time.Database Search and Analysis—Dta files were generated by Bio-works 3.2with default parameters and then treated by spectrum-filtering and spectrum optimization tools in pFind 2.1Studio.The candidate spectra of MS 3were searched against UniProt Knowledge-base Release 12.6(human,76,137entries;UniProt Knowledgebase Release 12.6consists of UniProtKB/Swiss-Prot Release 54.6of De-cember 4,2007and UniProtKB/TrEMBL Release 37.6of December 4,2007)using the pFind 2.1search engine.The database was modified by substituting the letter N in glycosylation sequence N X (S/T/C)with J,which was defined to have the same mass as Asn (21),and then the target and reversed decoy database were combined for the search.Carbamidomethylation was considered for all Cys residues.Variable modifications contained oxidation of Met residues,carbamidomethy-lation and carbamylation (carbamylation was only considered as a F IG .3.MS 2and MS 3spectra of fucosyl-GlcNAc-attached peptides.The peak intensity distribution of the MS 3spectrum is much more homogeneous than that of MS 2,so better peptide sequence information can be obtained;the direct assignment of CF glycosites can be deduced from the b-type and y-type ion series attached with a GlcNAc residue in MS 3.a and b are MS 2and MS 3spectra of GLC #VJASAVSR from insulin-like growth factor-binding protein 3,respectively.The peaks of b-type and y-type ions with or without GlcNAc residues appear synchronously and frequently,such as y 7ϩand b 6ϩ.c and d are MS 2and MS 3spectra of a candidate that was analyzed de novo ,respectively.The resulting de novo sequence GVEIJR (because the m /z of ion b 1is too low to detect,the sequence of the first two residues can also be “VG,”and “I”can also be “L”because of their same mass)was not found in the peptide database of tryptic digests (J located in the sequon N X (S/T/C)where X is any amino acid except proline).D 1,C 8H 14NO 5(GlcNAc);D 2,C 8H 12NO 4;D 3,C 8H 10NO 3;D 4,C6H 10NO 3;D 5,C 7H 8NO 2.The y 7G ϩidentifies the GlcNAc residue with the same sequence as y 7ϩ.J,CF site;#,carbamidomethylation.Precise and Large Scale CF Glycoprotein Identification916Molecular &Cellular Proteomics 8.5 by Xiaohong Qian on May 1, 2009 Downloaded fromvariable modification in experiments that used urea as the proteindenature reagent)of peptide N-terminal and Lys residues,and a 203.0794-Da variable addition to J residues.At most,two missed tryptic cleavage sites were allowed.Tolerance of parent ions wasϮ20 ppm,and tolerance of fragment ions wasϮ0.5m/z for the primary search.The final identified results had a1%false-positive rate(22), and the tolerance for parent ions wasϮ10ppm.MALDI-TOF MS Analysis—After desalting with the C18ZipTip,all of the samples were mixed1:9with5mg/ml␣-cyano-4-hydroxycin-namic acid in50%acetonitrile supplemented with0.1%TFA,and0.5l of sample was applied to the MALDI target plate.The mass spectra were obtained using a4800Proteomics Analyzer MALDI-TOF/TOF instrument(Applied Biosystems).Prior to analysis,the mass spec-trometer was externally calibrated with seven peptides obtained from tryptic digest of myoglobin.The m/z range of the MS scan was from 600to4000.Mass spectra were acquired in the positive reflector mode.RESULTS AND DISCUSSIONCore-fucosylated Glycopeptide Enrichment from Plasma—Robust and convenient operation procedures were estab-lished to obtain partially deglycosylated CF glycopeptides. After IgG depletion,plasma proteins were mixed with L.culi-naris lectin to enrich for the CF glycoproteins.Binding pro-teins were digested by trypsin,and the resulting glycopep-tides were enriched through a molecular weight cutoff technique.N-Linked glycopeptides usually have larger molec-ular weights than non-glycopeptides(19,23);therefore,an ultrafiltration membrane with a molecular mass limit of3000 Da was utilized to enrich for glycopeptides.This step inte-grates enrichment,desalting,and concentration into one op-eration.Glycopeptides were then treated with endoglycosi-dase F3,which specifically cleaves the glycosidic bond between the two proximal N-acetylglucosamines(GlcNAc) and leaves the fucosyl-GlcNAc residues on the peptides. Endoglycosidase F3was chosen here for treating CF glyco-protein because a large number of the glycans of plasma glycoproteins have biantennary structure,which is a moreefficient substrate for endoglycosidase F3(24).For other structures,such as tetraantennary and other bulky glycans, the reactivity of endoglycosidase F3is poor,so there may need to be additional evaluation to choose the proper glyco-sidase for other kinds of samples like tissue biopsies.A tryptic peptide mixture from four standard glycoproteins, apotransferrin,fetuin,rhEPO,and ribonuclease B,was used to illustrate the efficiency of the ultrafiltration method(Fig.1). Half of this tryptic peptide mixture was directly treated with peptide-N-glycosidase F(untreated sample);the other half was separated by ultrafiltration into a retentate fraction(high molecular weight)and a filtrate fraction(low molecular weight),and then both fractions were treated with peptide-N-glycosidase F.The deglycosylated glycopeptides were de-tected by theϩ0.984-Da mass drift on Asn to Asp.In total,eight N-glycopeptides were reported for four glycoproteins.Six of these glycopeptides were directly found in untreated samples by MALDI-TOF MS.However,in addition to these six glycopeptides,one more glycopeptide (CGLVPVLAENYN*K from apotransferrin;N*represents the annotated glycosite)was detected in the retentate fraction. The relative intensities of all deglycosylated glycopeptides were heightened compared with the untreated sample.In the untreated sample,the failure to detect CGLVPVLA-ENYN*K is ascribed to suppression by a non-glycopeptide with similar mass.In the filtrate fraction,the relative intensity of deglycosylated glycopeptides decreased to a very low level,illustrating that few glycopeptides were lost.One re-ported glycopeptide was not detected in the three fractions (N*LTK from ribonuclease B).One possible reason is that its sequence is too short to detect.Development of Neutral Loss-dependent MS3Scan Meth-od—A neutral loss-dependent MS3method specifically de-signed for partially deglycosylated CF glycopeptides was de-veloped.During CID,the glycosidic bond that links the two remaining sugars is prone to breakage compared with the other bonds(25).In our experiments on three partially degly-cosylated CF glycopeptides,the highest peaks in the MS2F IG.4.The process of the strategy for CF glycoprotein identi-fication.CF glycoprotein identification was achieved through enrich-ment of CF glycopeptides,partial deglycosylation of CF glycopep-tides,HPLC neutral loss-dependent MS3,candidate spectrumfiltering,spectrum optimization,and database searching.F1identifies the intensity ratio of the second strongest peaks(logogram:secondstrong peak(SSP),which does not contain different states for S2,such as a different charge state or states of H2O and NH3loss)to S2;F2identifies the difference between the calculated and experimentalm/z of S2;F3identifies the intensity ratio of the second strongest peak(logogram:SSPЈ)to S3within the range of the S3monoisotopic peak Ϯ3m/rmation on different charge state ions of S3is considered, and the better result is recorded.Additionally the absolute intensitiesof S2and S3are required to be higher than500and50,respectively. As shown,different scores correspond to different signal qualities. The confidence of the spectrum is sorted into five ranks by total score.Œ,fucose residue;f,GlcNAc residue.2D,two-dimensional; Endo,endoglycosidase;LCH,L.culinaris lectin.Precise and Large Scale CF Glycoprotein IdentificationMolecular&Cellular Proteomics8.5917 by Xiaohong Qian on May 1, 2009 Downloaded fromspectra all resulted from subtraction of 146Da (massof the fucose residue)from the parent ions that had the same charge state as the corresponding parent ions (Fig.2).Based on this trait,a neutral loss-dependent MS 3scan method was utilized as an automatic event in the LTQ-FT mass spectrometer:MS 3spectra were automatically collected when one of the three most intense peaks from the MS 2spectrum corresponded to a neutral loss event of the fucose residue mass.MS 3spectra were generated from fragmentation of the GlcNAc-attached pared with the MS 2spectra,which were gen-erated from fragmentation of the fucosyl-GlcNAc-attached peptides,the MS 3spectra have three remarkable advantages.1)They have better spectrum quality:the peak intensity distribution of the MS 3spectrum is much more homogene-ous.This is beneficial because there are more fragment ion signals with good signal to noise ratios.2)They have sim-pler spectrum information:the number of theoretical frag-ment ions in the MS 3spectrum is fewer.This makes the algorithm for peak matching simpler and easier.3)They have clearer spectrum signals:two parent ion selections (from MS to MS 2and from MS 2to MS 3)reduce the proba-bility of collecting interference signals adjacent to parent ions in the full scan (Fig.3).In addition,direct assignment of CF glycosites can be deduced from the b-type and y-type ions series attached with a GlcNAc residue,providing much higher confidence levels of glycosite assignment compared with the 0.984-Da mass shift method.It should be noted that the retained intact GlcNAc residues were found to be lost from the b and y ions (Fig.3);therefore,these kinds of special product ions must be considered in addition to GlcNAc attached b and y ions when searching the data-base.This observation was taken into account for peptide scoring in the pFind 2.1search engine (26–28).Compared with other popular software tools,pFind discovered more results (supplemental Data 1).Development of Candidate Spectrum-filtering and Spec-trum Optimization Methods—Due to the complexity of real samples and the massive spectra generated in these large scale glycopeptide analyses,more professional and special-ized processing methods are absolutely necessary.Here a database-independent method for discovery of spectra of partially deglycosylated CF glycopeptides was developed.Two kinds of ions in MS 2were scrutinized and used to judge whether the precursor was a CF glycopeptide:ions of a peptide attached to a GlcNAc residue (symbol ion 2,logo-gram:S 2,attained from the breakage of the glycosidic bond between the remaining two monosaccharide residues)and ions of a pure peptide (symbol ion 3,logogram:S 3,obtained from fragmentation between the GlcNAc and the Asn residue of the peptide).By introduction of the highly accurate parent ion mass from a full scan (recorded in FT-ICR),we can cal-culate the m /z of symbol ions.Next according to the quality of the symbol ions in MS 2,several criteria were established to sort out the spectra.First of all the strongest peak in MS 2F IG .5.Frequency histogram of in-tact and partial GlcNAc loss peaks incandidate MS 3spectra of charge 2.The m /z values of S 2were set as 0m /z .Offsets with high peak frequencies re-veal potential masses of neutral lossesthat frequently occur on peptide-at-tached GlcNAc residues.The possibleloss groups are shown in the table .Precise and Large Scale CF Glycoprotein Identification918Molecular &Cellular Proteomics 8.5 by Xiaohong Qian on May 1, 2009 Downloaded from。