“Yang Wei Kang Liu” improves the survival of late stage gastric cancer patients
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第1课人体的内环境与稳态高效演练·创新预测预测点1.内环境的组成成分1.医院里给病人输液时使用的生理盐水与血浆、组织液的渗透压相同,称为等渗溶液,但是过多注射会影响细胞的正常功能,下列叙述错误的是( )A.生理盐水与细胞外液虽为等渗溶液,但是成分差别非常大B.生理盐水成分与血浆成分明显不同,过量补充会影响pH的稳态C.过量补充生理盐水,血浆中的其他成分,如钾离子的浓度会偏低D.过量注射生理盐水会导致内环境渗透压发生改变【解析】选D。
生理盐水的主要成分是0.9%的NaCl溶液,不含蛋白质、糖类等物质,A正确;过量补充生理盐水会使内环境中的H+、K+浓度降低,B、C正确;生理盐水与血浆、组织液是等渗溶液,不改变内环境的渗透压,D错误。
2.机体通过内环境与外界进行物质交换,维持正常生命活动的进行,据图分析下列有关人体内环境的叙述,)A.乳酸与碳酸氢钠作用生成乳酸钠和碳酸的过程发生在①中B.图中④是细胞生存的液体环境,又称细胞外液C.图中③与①最主要的区别是③中的蛋白质含量较少D.图中②是血浆,内含葡萄糖、血浆蛋白和激素等成分【解析】选D。
根据题图判断①为血浆,②为组织液,③为淋巴。
乳酸与碳酸氢钠作用生成乳酸钠和碳酸,有利于维持血浆pH的稳定,A正确;内环境包括血浆、组织液和淋巴,B正确;内环境中,血浆中的蛋白质含量最高,C正确;②是组织液,且组织液中不含血浆蛋白,D错误。
预测点2.内环境的理化性质3.九寨沟地震后,多名受困、受伤人员需要救助,下列关于被救助人员内环境及稳态的说法中,正确的是( )导学号14120159A.受困人员被解救后,由于饥饿而摄入过多、过咸食物后,会引起细胞内液的量增加B.由于缺氧,受困人员的骨骼肌纤维内乳酸积累过多,会引起细胞体积增大C.给受困、受伤人员静脉滴注0.9%的NaCl溶液(生理盐水)后,会使细胞内液的渗透压降低D.受伤人员血浆蛋白减少使血浆渗透压升高,可出现组织水肿【解析】选B。
英语医疗保健英语30题1. The doctor asked the patient to lie _____.A. downB. upC. inD. on答案:A。
本题考查短语“lie down”,意思是“躺下”。
选项B“lie up”不存在此短语;选项C“lie in”意思是“在于”;选项D“lie on”意思是“位于,压迫”。
在医疗场景中,医生让病人躺下常用“lie down”。
2. You should take the medicine _____ a day.A. two timesB. twiceC. secondD. two答案:B。
“twice a day”表示“一天两次”,是固定用法。
选项A“two times”表述不太准确;选项C“second”是序数词“第二”;选项D“two”是基数词“二”,都不符合表达“一天两次”的意思。
3. The nurse is very kind. She always takes good care _____ the patients.A. ofB. forC. atD. to答案:A。
“take care of”是固定短语,意思是“照顾”。
选项B“take care for”不存在此搭配;选项C“take care at”和选项D“take care to”也不是正确的表达。
4. After the operation, the patient needs to have a _____ rest.A. goodB. badC. wellD. ill答案:A。
“have a good rest”表示“好好休息”,是常用表达。
选项B“bad”是“坏的”,不符合语境;选项C“well”作形容词时表示“身体好”,不能用于“have a well rest”;选项D“ill”是“生病的”,也不符合。
5. The doctor told him to give up _____.A. smokeB. to smokeC. smokingD. smokes答案:C。
八年级英语医疗救护单选题50题1. In the hospital, the doctor said that the patient had a pain in his ____. It might be a sign of appendicitis.A. stomachB. heartC. lungD. liver答案:A。
解析:本题考查身体器官的词汇。
题干提到可能是阑尾炎的症状,阑尾炎通常是腹部疼痛,stomach是胃、腹部的意思。
heart是心脏,lung是肺,liver是肝脏,这三个器官与阑尾炎的疼痛部位无关,所以A正确。
2. The nurse told the patient to take some ____ to reduce the fever.A. vitaminsB. antibioticsC. painkillersD. sleeping pills答案:C。
解析:本题考查治疗手段相关的词汇。
题干说为了退烧,vitamins是维生素,主要用于补充营养,不是用于退烧;antibiotics 是抗生素,用于抗感染,不是直接退烧;painkillers是止痛药,有退烧的作用;sleeping pills是安眠药,与退烧无关,所以C正确。
3. The doctor examined the patient's ____ carefully to see if there was something wrong with it.B. earC. noseD. eye答案:A。
解析:本题考查身体器官词汇。
根据题意是医生仔细检查某个器官看是否有问题,在这四个选项中,检查喉咙是否有问题比较常见,ear耳朵、nose鼻子、eye眼睛虽然也可检查,但从常见医疗场景来看,喉咙更符合题意,所以A正确。
4. A man was sent to the hospital because he had a serious ____. He could hardly breathe.A. coldB. headacheC. asthmaD. toothache答案:C。
中考英语医疗技术进步影响单选题40题1. The development of medical technology has greatly improved the treatment of cancer. Now, more and more patients can ______ the disease.A. survive fromB. surviveC. liveD. keep alive答案解析:B。
本题考查动词的用法。
选项A“survive from”这种用法错误,“survive”本身就可表示“从……中幸存,幸免于”。
选项B“survive”表示“幸存,存活”,在这里表示更多病人能在癌症这种疾病下存活,是正确的。
选项C“live”表示“生活,居住”,侧重于生活的状态,而不是在疾病下存活,所以不合适。
选项D“keep alive”表示“使……活着”,这里主语是病人,不是使别人活着,所以不符合语境。
2. With the progress of medical technology, the treatment of heart disease is no longer as difficult as before. Doctors can ______ many effective methods to save patients.A. useB. makeC. createD. find答案解析:A。
本题考查动词词义辨析。
选项A“use”使用,医生可以使用很多有效的方法来挽救病人,符合语境。
选项B“make”制作,这里不是制作方法,而是使用已有的方法,所以不正确。
选项C“create”创造,侧重于创造出原来没有的东西,这里强调使用现有的方法,所以不合适。
选项D“find”找到,找到方法并不意味着就能用来治病,重点是使用这些方法,所以该选项错误。
3. In recent years, medical technology has made great strides in treating diabetes. New drugs can help patients ______ their blood sugar levels better.A. controlB. keepC. manageD. all of the above答案解析:D。
2014年中考英语主谓一致复习题(有答案含13年中考题)涓昏皳涓€鑷?( )1.(2013骞撮粦榫欐睙榛戞渤)Summer holiday is coming, Li Lei with his father________to go to Shanghai. A.want B.will want C.wants ( )2.(2013骞撮粦榫欐睙缁ュ寲)Not only my father but also my grandpa ________flying to Singapore this weekend. A.is B.are C.was ( )3.(2013骞撮粦榫欐睙缁ュ寲)鈥昗hich would you like, tea or coffee? 鈥昣_______is OK, I don't mind. A.Either B.Both C.Any ( )4.(2013骞撮粦榫欐睙缁ュ寲)A number of students ________ playing on the playground, and the number of students in our school ________ increasing. A.are; are B.is; are C.are; is ( )5.(2013骞存箹鍖楅粍鐭?________ that pair of ________ a little ch eaper? A.Is; glass B.Are; glass C.Is; glasses D.Are; glasses ( )6.(2013骞存箹鍗楀父寰?鈥昗hat's on the desk? 鈥昑here ________ some books. A.be B.are C.is ( )7.(2013骞存睙鑻忓崡閫?Each of the club members ________ready to help those who were in trouble. A.is B.are C.was D.were ( )8.(2013骞存睙鑻忔嘲宸?鈥昗hy are you late, Jim? 鈥旴ecause there ________ a lot of traffic when I came here. A.is B.are C.was D.were ( )9.(2012骞翠笂娴?There ________ still some milk in the fridge.It's not necessary to go to the store today. A.am B.is C.are D.be ( )10.(2012骞村北涓?鈥昗hat can you see in the picture? 鈥旾can see a farm.And there________ a lot of animals on it. A.is B.are C.will be D.be ( )11.(2012骞撮粦榫欐睙榻愰綈鍝堝皵)This is my twin sister, Lucy.Not only she but also I ________ good at drawing. A.is B.am C.a re ( )12.(2012骞村洓宸濆箍瀹?鈥旽ow much is the pair of shoes? 鈥昑wenty dollars ________ enough. A.is B.are C.am ( )13.(2012骞村洓宸濆痉闃?鈥昉hysics ________ more difficult than Chinese, do you think so? 鈥昚es, I think so. A.is B.are C.has ( )14.(2012骞村洓宸濊祫闃?Each of the girls here________ to the West Lake twice. A.have gone B.have been C.has gone D.has been ( )15.(2012骞磝鐤嗕箤椴佹湪榻?There ________ going to be a basketball matchthis afternoon. A.are B.is C.have D.will ( )16.(2012骞存箹鍖楅粍鐭?The head teacher with his students ________ Tuanchengshan Park if it ________ tomorrow. A.is going to; isn't rainy B.are going to; isn't rainy C.is going to; won't rain D.are going to; doesn't rain ( )17.(2012骞村洓宸濈坏闃?The number of books in the bookshop is about 10,000 and a number of them ________ about science. A.is B.was C.areD.were ( )18.(2012骞磋吹宸為摐浠?Neither my father nor my mother ________ rock music.They think that it's too ________. A.like; noise B.likes; noise C.like; noisy D.likes; noisy ( )19.(2012骞存睙鑻忚嫃宸?There are millions of websites on the Internet and there________a lot of useful________on the websites. A.are; informations B.are; information C.is; information D.is; informations ( )20.(2012骞存箹鍗楄闃?鈥旸o you like your new T卢shirt? 鈥昚es.Not only I but only my mother ________ it. A.likes B.like C.doesn't like ?( )1.Every student and every teacher ________ looking forward to the culture festival. A.be B.are C.to be D.is ( )2.To swim with you ________ very enjoyable. A.is B.are C.become D.to be ( )3. (2014鍘熷垱)Nothing ________ impossible. A.are B.is C.were D.to be ( )4.This pair of glasses ________ to Tom. A.belong B.is C.belongs D.are ( )5.The kind卢hearted________ welcome everywhere. A.is B.are C.was D.is to ( )6.Generally, common people ________ to see light卢hearted movies. A.like B.likes C.goes D.going ( )7.Over 50% of the fresh water in the world ________ in the polar areas. A.are B.is C.were D.was ( )8.鈥旾have two shirts for you.Do you like them? 鈥昇either________ suitable for me, I think. A.are B.has C.is D.have ( )9.Neither Steve nor his brothers ________ red shoes. A.is B.are C.have D.has ( )10.Our Chinese teache r as well as all of us ________ going to the park tomorrow. A.are B.will C.was D.is( )11.There ________ a pen and some books on the table. A.are B.is C.has D.have ( ) 12.Jane with her parents ________ at home. A.stay B.leave C.stays D.leaves ( )13.The police ________ that the criminal ________ on the run. A.report; is B.report; are C.reports; is D.reports; are ( )14.Mathematics ________ my favorite.And I likegeography too.Bot h ________ my strong subjects. A.is; is B.are; are C.are; is D.is; are ( )15.Ou r class ________ made up of 25 boys and 30 girls. A.is B.are C.has D.have ( )16.Six months ________ too long for a child living alone. A.are B.is C.wait D.waits ( )17.The singer and actor ________ a wonderful performance. A.give B.gives C.is D.are ( )18.There ________ some water and some seeds in the bottle.The seeds________ very small. A.are; is B.are; are C.is; are D.is; is ( )19.No one who can play well ________ here now.Either of you ________ the chance to win. A.is; has B.is; have C.are; has D.are; have ( )20.Both of you ________ good at swimming, but we just need one more person.Either he or you ________ with me. A.is; come B.is; comes C.are; come D.are; comes ?( )1.A library with five thousandbooks________to the nation as a gift. A.is offered B.has offered C.are offered D.have offered ( )2.When and where to build the new factory________ yet. A.is not decided B.are not decided C.has not decided D.have not decided ( )3.The number of people invited________fifty, but a number of them________absent for different reasons. A.were; was B.was; was C.was; were D.were; were ( )4.Between the twobuildings________a monument. A.is standing B.standing C.stands D.stand ( )5.Many a student ________that mistake before. A.had made B.has been made C.have made D.has made ( )6.None of the money________his. A.is B.are C.belongs D.were ( )7.About three卢fifths of the work________done yesterday. A.had B.was C.were D.have ( )8.Neither your sister nor mine________the good news.Let's tell them.A.knowB.knowsC.knewD.have known ( )rge quantities ofwater________cooling purposes. A.are needed for B.is needed to C.are needed to D.is needed for ( )10.Every boy and every girl________to attend the evening party.A.wishB.wishesC.hopeD.are hopping ( )11.Everything ________ in order. A.is B.are C.has D.have ( )12.Mary as well as her sisters________Chinese in China. A.are studying B.have studied C.studies D.study ( )13.Not only I but also Jane andMary________tired of having one examination after another. A.is B.are C.am D.be( )14.I don't think the poor________poor. A.are always B.is always C.always beD.always ( )15.How time flies锛乀hree years ________ really a short time. A.was B.is C.were D.are ( )16.Both of us ________to the party last week. A.was invited B.were invited C.invited D.is invited ( )17.Interest, as well as prospects, ________important when one looks for a job. A.are B.were C.is D.was ( )18.A number of cars________in front of the park. A.is par ked B.was parked C.are parked D.has parked ( )19.The number of articles published on smoking ________ amazing. 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小学上册英语第三单元真题试卷英语试题一、综合题(本题有100小题,每小题1分,共100分.每小题不选、错误,均不给分)1.Hydrogen is the lightest ______.2.He is a fast ___. (runner)3.Chlorine is commonly used as a _____ to disinfect water.4.The _______ of a plant can be very strong.5.What is the color of a ripe banana?A. GreenB. YellowC. RedD. Brown6. A __________ is a place that is protected by law.7.What do you call a person who helps people find their way?A. GuideB. NavigatorC. LeaderD. PathfinderA8. A ______ (生态平衡) is vital for sustaining life.9. A __________ is a small area of land surrounded by water.10.The first country to grant women the right to vote was _______.11.What is the name of the event where you give thanks for what you have?A. ChristmasB. HalloweenC. ThanksgivingD. New YearC12.The Nile River flows through __________.13.What do we call a person who studies rocks?A. BiologistB. ChemistC. GeologistD. Physicist14.The chemical symbol for lead is ______.15.The _______ (兔子) has a very soft coat.16.I like to eat _____ (apples).17.I help my sister with her __________. (作业)18.We have a ______ (快乐的) reunion every year.19.What is the main function of the heart?A. BreathingB. Pumping bloodC. DigestionD. Thinking20.The ________ was a crucial chapter in the narrative of national unity.21.What do we call the study of viruses and bacteria?A. VirologyB. MicrobiologyC. PathologyD. Immunology22.The ________ is very wise and watches over us.23.We have a ______ (丰富的) library collection.24.In _____ (摩洛哥), you can visit the Atlas Mountains.25.The symbol for dysprosium is _____.26.The koala is native to ________________ (澳大利亚).27.What do we call a young adult?A. ChildB. TeenagerC. SeniorD. BabyB28.The ______ is the center of an atom.29.The jellyfish's tentacles can capture ________________ (猎物).30.An exothermic reaction ______ energy to the surroundings.31. A __________ is a large body of freshwater.32.The ______ (花园) is full of ______ (昆虫) in summer.33.My favorite toy can ________ (动词) in many ways, which makes it special.34. A ________ (湿润地区) supports diverse wildlife.35.I can ___ (spell) my name.36.Sodium chloride is commonly known as _____.37. A _______ can measure the energy output of a solar panel.38.What do you call the time it takes for the Earth to rotate once?A. YearB. MonthC. DayD. HourC39.What is the name of the famous tower in Paris?A. Big BenB. Leaning Tower of PisaC. Eiffel TowerD. Tower of London40.The dog is ________ (跟我走).41.What do we call the lines that run north to south on a map?A. LatitudeB. LongitudeC. EquatorD. Meridian42.The __________ (气象) often changes quickly.43.I enjoy _____ (参加) local events.44.I can _____ (游泳) well.45.My favorite character in a TV show is _______ (名字). 他/她很 _______ (形容词).46.My dad loves to cook __________. (晚餐)47.The kitten likes to chase a _________. (线)48.The __________ (历史的反思) leads to personal growth and development.49.The chemical formula for ammonium nitrate is _______.50.I can ______ (为) my family meals.51. A dwarf planet is similar to a planet but does not clear its orbit of ______.52.I often tell my friends, "You can call me ______ anytime!" (我经常告诉我的朋友:“你随时可以叫我!”)53.Baking soda is a common _____ used in cooking.54.The _____ (纸) is white.55.Chemical reactions are often represented by ________ equations.56.Which of these is a mode of transportation?A. TrainB. ChairC. TableD. Lamp57.Plants absorb sunlight to perform _______. (光合作用)58.What do we call the study of living things?A. BiologyB. ChemistryC. PhysicsD. AstronomyA59.I enjoy playing ______ (棋盘游戏) with my family during the weekends.60.My favorite _____ is a red fire truck.61.What do we call the process of a caterpillar turning into a butterfly?A. MetamorphosisB. EvolutionC. TransformationD. DevelopmentA62.What do we call the process by which plants absorb water through their roots?A. TranspirationB. AbsorptionC. EvaporationD. Filtration63.The __________ (历史的记忆保留) ensures continuity.64.The ________ is green and leafy.65.The tarantula is often feared for its ______ (外表).66.We will have a ______ tomorrow at school. (presentation)67.The _____ (clover) is a symbol of luck.68.I enjoy _______ (看书) at the library.69. A _____ (小猫) loves to chase after strings.70. A ______ is a type of insect that can be very colorful.71.She likes to ________ books in the library.72.The tree is _______ (swaying) gently.73.The __________ is a region known for its fishing.74.What is the name of the famous superhero who is also a scientist?A. The HulkB. Iron ManC. ThorD. Captain AmericaB75.Plants need __________ (阳光) to grow well.76.What is the name of the famous American president who abolished slavery?A. George WashingtonB. Abraham LincolnC. FranklinD. RooseveltD. Thomas Jefferson77.I have _____ of friends in my class. (lots)78.What is the primary color that, when mixed with red, creates pink?A. BlueB. YellowC. WhiteD. GreenC79.ts have ______ (刺). Some pla80.They are going to the _____ (公园).81.an Revolution led to the establishment of the ________ (苏维埃政权). The Russ82.What do we call the area of land that is home to a specific species?A. HabitatB. EcosystemC. BiomeD. NicheA Habitat83.What is the main purpose of a museum?A. To entertainB. To educateC. To sell goodsD. To provide shelterB84.The __________ (历史的回响) resonates through time.85. A _______ can be used to measure the density of a solid object.86. A __________ is a measurement of how much matter is in an object.87.I have _______ (两只) cats at home.88.My favorite animal is a ________ that loves to eat.89. A seal's blubber keeps it warm in ________________ (寒冷) waters.90.What is the capital of Russia?A. MoscowB. St. PetersburgC. KazanD. VladivostokA91.Bees help ______ plants by pollinating.92.How many continents are there in the world?A. 5B. 6C. 7D. 893.The atomic structure of an element determines its ______.94. A ________ (北极熊) lives in the cold Arctic and is very large.95.The shark is very ___ (fearsome).96.I like to ______ (学习) about animals.97.What do bees collect from flowers?A. NectarB. HoneyC. PollenD. Water98.What is the largest planet in our solar system?A. EarthB. MarsC. JupiterD. Saturn99.I enjoy telling stories with my toy ________ (玩具名称). 100.My ________ (玩具名称) is a delightful companion.。
小学四年级英语下册全册教学设计(北师大版)Unit 12 ReviewLesson 1 Preparation You will need: lashcards for the vocabulary being reviewed in Units 7 through 11. 1.Structure review: it鈥檚o (red) train; his, her, my old up the flashcard for train. Ask, 鈥淲hat鈥檚this?鈥?Elicit, 鈥淚t鈥檚a train.鈥?Ask, 鈥淚s it a big train?鈥?The children should answer, 鈥淵es, it is.鈥?sk what other words they can use to describe it. Elicit words like old, new, small, long, red, and yellow, in a sentence, for example, 鈥淚t鈥檚an (old) train.鈥? ollect some classroom objects from the children (books, erasers, pencils, pens, rulers). Hold up one object and have the children make a sentence to describe it. For example, 鈥淚t鈥檚a (new) (book).鈥?Repeat the procedure with the other items.hen returning the items to the children, ask, 鈥淲hose pen is it?鈥?Elicit, 鈥淚t鈥檚(her) pen.鈥?Repeat the procedure with other items., asking children by name to ensure the use of their. For example, 鈥淭hey鈥檙e (their) books.鈥? 2.Set the scene ay, 鈥淲hat do we say to someone if we don鈥檛want them to do something?鈥?Elicit the term in English, Don鈥檛! resent the flashcard for ticket. Model the word and write it on the board. Have the children repeat the word after you. sk the children what they say when they are at the train station and want to buy some tickets. Try to elicit, 鈥淚want some tickets, please.鈥?or鈥淗ow much are the tickets?/How much is the fare?鈥?All of these are correct. 3.Talk about the story Student Book pages 62 and 63Have the children open their books at pages 62 and 63. Ask these questionsabout the pictures: Picture 1: 鈥淲hat do you think Danny is asking Bobby?鈥?Picture 2: 鈥淲hat are the friends waiting for?鈥?銆€銆€銆€銆€銆€銆€銆€銆€鈥淲hat color is the train?鈥?Picture 3: 鈥淲hat color is this train?鈥?鈥淚s this the train the friend are waiting for?鈥?Pictures 4/5: 鈥淲hat is Mocky looking at?鈥?Pictures 6/7/8: 鈥淲hat is happening now?鈥?Picture 9: 鈥淲hat is Mocky doing?鈥?Picture 10: 銆€鈥淚s Mocky better now?鈥?4.Story Student Book pages 62 and 63 ay, 鈥淣ow we will hear what the characters said.鈥?Play the tape and have the children look at the pictures as they listen. lay the tape again, pausing at each picture. Have the children repeat the words each time. xplain that in English people say Ouch! when they are hurt. 5.Flashcard game This game reviews the core vocabulary introduced in Units 7 through 12. You will need the flashcards for the vocabulary being reviewed. Do not use the flashcards for numbers or action verbs. ivide the class into two teams. Put all the flashcards face down on a table between the teams. ave a child in Team A pick up a card and show it to Team B.(The children should take turns picking up the cards.) The child should ask, 鈥淎re these (dolls)?/Is this a (train)?鈥漮r 鈥淲hat鈥檚this?/What are these?鈥?ogether, Team B must answer, 鈥淭hey鈥檙e (doffs).鈥?or 鈥淚t鈥檚a (train).鈥?oints are scored for every correct question and correct answer. Lesson 2 Preparation You will need: pposites Master 1.Review Ensure that each child has his/her copy of the Opposites Master. 鈥how your own copy and point to the picture of the young turtle. Ask, 鈥淚s this a young turtle?鈥?The children should answer, 鈥淵es, it is.鈥?Now ask, 鈥淚s this an old turtle?鈥?Elicit, 鈥淣o, it isn鈥檛.鈥?ontinue for the pictures of the birds (big/small) and boy/girl(tall/short). If necessary, repeat the procedure until all the children answer each question correctly.2.Listen for words Student Book page 64 ave the children open their books and look at the picture of Ken and Mocky at the top of the page. Ask, 鈥淲here are Ken and Mocky?鈥?Elicit the answer, 鈥淭hey are in a store.鈥?Then ask, 鈥淲ho is the storekeeper?鈥?The answer is Mocky. ave the children look at the speech bubbles in the picture, and read the words to them. Explain that Ken is asking ifMocky has the things listed on the right銆€hand side. lay the first dialog and stop the tape. Ask, 鈥淒oes Mocky have a small train?鈥?The answer is Yes. 3.Listen for sounds Student Book page 64 irect the children鈥檚attention to the bottom of the page. Show your copy of the page and point to the first picture. Say the word goat and have the children repeat it after you. Continue for the other five pictures. lay the first word on the tape and have the children point to the matching picture. Repeat for the other words. lay the tape again. This time have the children write the number of the word in the box next to the matching picture. 4.Read, trace and match Student Book page 65 how your copy of the page. Point to the picture of the hamburger and ask, 鈥淲hat鈥檚this?鈥?The children should answer, 鈥淚t鈥檚a hamburger.鈥?Then have the children find the matching phrase. epeat the procedure for all the items on the table.ead each phrase once more. This time have the children draw a 1ine connecting each phrase to its correct picture. 5.Set homework Student Book page 65Explain that you want the children to color the pictures. Lesson3Preparation You will need: hildren's cards for Unit 7 through 12. 1.Talk together Student Book page 66 ave the children turn to page 66. Draw their attention to the top of the page. Show your copy of the page. Point to the first picture and ask,鈥淲ho bought the tickets in the story?鈥滶licit,鈥淒anny.鈥漈hen ask,鈥淲ho sold the tickets?鈥滶licit,鈥淏obby.鈥?odel the sentences in the first dialog and have the children repeat the sentences after you. ivide the class into two groups and have the children repeat the dial a roleplay. (One group is Danny and the other group is Bobby.) ut the children in pairs and have them practice the dialog. Encourage them to substitute different numbers of tickets. or the second dialog, point to the picture and ask, 鈥淲hat animials did Mocky see from the train?鈥?Then repeat the procedure used for the first dialog. (Three groups are needed this time.) Encourage the children to substitute different animals and animalnoises. For example, substitute small cats for big cows, and meow, meow for mooooo, mooooo or the third dialog, point to the picture and ask, 鈥淲hat happened to Mocky?鈥?Then repeat the procedure used for the second dialog. (Divide the class into three groups.) Have the children substitute differert body parts. The child who plays Mocky should point to the part of the body that is hurt. For example, have the child point to an ear when he or she says, 鈥淢y ear hurts.鈥?2.Review words ave a child come to the front of the class. Point to his or her ear and ask, 鈥淲hat鈥檚this?鈥?The class should answer,鈥淚t鈥檚(her) ear.鈥?epeat the procedure for the other parts of the body listed in the margin of this page. 3.Write words Student Book page 66 irect the children鈥檚attention to the bottom of page 66. Show your copy of the page and point to the eye. Say, 鈥淲hat鈥檚this?鈥?Try to elicit, 鈥淚t鈥檚her eye.鈥?Write the word on the board. Have the children copy the word on the correct line in their books. epeat the procedure for the remaining words. 4.Listen and circle Sudent Book page 67 ave the children open their books at page 67. Show your copy of the page and point to the first row of pictures. Tell them that only one word has a different/c/sound in the first three words. lay the tape for the first three words. Have the children repeat. Say,鈥淲hich word has the different/c/sound?鈥?Have them touch the word on their books. lay the tape again this time have the children circle out the word that has the dieffrent /c/ sound. 5.Snap! Use this game to review the vocabulary introduced in this book. It is best played in pairs. ave the children take out their cards for Units 7锝?2. ee Unit 8, page 50, for the rules and procedure. Lesson 4 1.Structure review:a/some ivide the class into two teams. Put all the food and drink flashcards for Unit 9 face down on a table between the teams. ave a child in Team A pick up a card and show it to Team B. The child should ask, 鈥淲hat鈥檚this?鈥漈ogether, Team B must answer, 鈥淎(hamburger).鈥?or 鈥淪ome (rice).鈥?Then the child in Team A asks, 鈥淚s this (hamburger) hot?鈥?The children inTeam B should answer, 鈥淵es, it is.鈥?or鈥淣o, it isn`t.鈥?ave the teams take turns at asking and answering the questions until most children have asked a question. Points are scored for each correct question and answer.2.Structure review: Is he/she (eating) (some soup)? eep the class in the two teams. Explain that this time, Team A will choose an eating and drinking activity, for example, eating hot soup. One child in the team mimes eating hot soup, and the other members in the team answer questions from Team B. he members of Team B must try to guess the activity. They could ask, 鈥淚s (she) drinking milk?鈥漈eam A should answer, 鈥淣o, (she) isn`t.鈥?hen Team B has guessed the activity, they must find out if the food is hot or cold. They should ask,鈥淚s the soup hot?鈥漈eam A should answer, 鈥淵es, it is.鈥?ave the teams change roles after an activity has been correctly guessed. emind the children of other food words they have learned. oints are scored for each correct question and answer, and for correctly guessing the activity. 4.Let鈥檚sing Student Book page 68ave the Student open their books at page 68. Draw their attention to the picture. Ask them to talk about the picture. You can ask questions like 鈥淲hat are the children doing?鈥? 鈥淎re they happy?鈥? etc. epeat the words, aloud to the children. Have them repeat after you. lay the tape straight through. Have the children clap their hands with the tape. lay the tape again. This time encourage them to join in the song. lay the tape. Have the children stand in a circle and walk up and down in the circle with the music. 5. Set homework Student Book page 68 Tell the children to sing the new song to their families.。
TheMidnightVisitorThe Midnight VisitorAusable did not fit the description of any secret agent. Fowler had ever read about. Following him down the corridor of the old French hotel where Ausable had a room, Fowler felt disappointed. It was a small room, on the sixth and top floor and hardly a setting for a romantic figure.銆?銆? Ausable was, for one thing, fat. Very fat. Very fat. And then there was his accent. Though he spoke French and German fairly well, he had never altogether lost the New England accent he had brought to Pairs from Boston twenty years ago.銆?銆? 鈥淵ou are disappointed,鈥?Ausable said over his shoulder. 鈥淵ou were told that I was a secret agent, a spy. You wished to meet me because you are a writer, young and romantic. You expected mysterious figures in the right, the crack of pistols, drugs in the wine.鈥?銆?銆? 鈥淚nstead, you have spent a whole evening in a French music hall with a dirty顎憀ooking fat man who, instead of having messages slipped into his hand by dark -- eyed beauties, gets only an ordinary telephone call making an appointment in his room. You have been bored!鈥?The fat man laughted quietly as he unlocked the door of his room and stood aside to let his guest enter.銆?銆? 鈥淏ut take cheer, my young friend,鈥?Ausable told him.銆?銆? 鈥淪oon you will see a paper, a quite important paper for which several men and women have risked their lives, come to me in the next顎憈o顎憀ast step of its journey into official hands. Some day that paper may well affect the course of history.In that thought is drama, is there not?鈥?As he spoke, Ausable closed the door behind him. Then he turned on the light.銆?銆? And as the light came on, Fowler had his first shock of the day. For halfway across the room, a small pistol in his hand, stood a man. Ausable blinked a few times.銆?銆? 鈥淢ax,鈥?he said, 鈥測ou gave me quite a start. I thought you were in Berlin. What are you doing in my room?鈥?銆?銆? Max was thin, not tall, and with a face that suggested the look of a fox. Except for the gun, he did not look very dangerous.銆?銆? 鈥淭he report,鈥?he said in a quiet voice. 鈥淭he report that is being brought to you tonight about some new missiles. I thought I would take it from you. It will be safer in my hands than in yours.鈥?銆?銆? Ausable moved to an armchair and sat down heavily. 鈥淚'm going to raise the devil with the management this time; I am angry,鈥?he said firmly. 鈥淭his is the second time in a month that somebody has gotten into my room from that balcony!鈥?Fowler's eyes went to the single window of the room. It was an ordinary window, with the black night outside.銆?銆? 鈥淏alcony?鈥?Max asked. 鈥淣o, I had a passkey. I did not know about the balcony. It could have saved me some trouble had I known about it.鈥?銆?銆? 鈥淚t's not my balcony,鈥?explained Ausable angrily. 鈥淚t belongs to the next apartment. You see, this room used to be part of a large unit, and the next room had the balcony, which extends under my window now. You can get onto it from the empty room next door, and somebody did, last month. The management promised to block it off. But they haven't.鈥?銆?銆? Fowler was standing stiffly near Ausable. 鈥淧lease sitdown,鈥?said Max to Fowler, waving his gun with a commanding gesture. 鈥淲e have a wait of about half an hour.鈥?銆?銆? 鈥淚 wish I knew how you Germans learned about the report, Max,鈥?said Ausable.銆?銆? The little spy smiled. 鈥淎nd we wish we knew how people got the report. But, no harm has been done. I will get it back tonight. What is that? Who is at the door?鈥?銆?銆? Fowler jumped at the sudden knocking at the door.銆?銆? usable just smiled. 鈥淭hat will be the police,鈥?he said, 鈥淚 thought that such an important paper as the one we were waiting for should have a little extra protection. I told them to check on me to make sure everything was all right.鈥?銆?銆? Max bit his lip.The knocking was repeated.銆?銆? 鈥淲hat will you do now, Max?鈥?Ausable asked. 鈥淚f I do not answer the door, they will enter anyway. The door is unlocked. And they will not hesitate to shoot.鈥?銆?銆? Max's face was black with anger as he backed swiftly toward the window; with his hand behind him, he opened the window and put his leg out into the night. 鈥淚 will wait on the balcony. Send them away or I'll shoot and take my chances!鈥?銆?銆? The knocking at the door became louder and a voice was raised. 鈥淢r. Ausable! Mr. Ausable!鈥?銆?銆? Keeping his body twisted so that his gun still covered the fat man and his guest, the man at the window seized the frame with his free hand to support himself as he rested his weight on one thigh. Then he swung his other leg up and over the window sill.銆?銆? The doorknob turned. Swiftly Max pushed with his left hand to free himself, and dropped to the balcony. And then as he dropped, he screamed once shrilly.銆?銆? The door opened and a waiter stood there with a tray, a bottle and two glasses. 鈥淗ere is the drink you ordered, sir.鈥?He set the tray on the table and left the room.銆?銆? White faced and shaking, Fowler stared after him. 鈥淏ut... but... what about... the police?鈥?he stammered.銆?銆? 鈥淭here never were any police,鈥?Ausable said. 鈥淥nly Henry, whom I was expecting.鈥?銆?銆? 鈥淏ut what about the man on the balcony...?鈥?Fowler began.。
2014年八年级下册英语8B期中测试题(有答案)2013-2014瀛﹀勾搴?B鏈熶腑娴嬭瘯鍗?涓€銆佸崟椤归€夋嫨锛?)1. 鈥旾have _______finished my homework. What about you? 鈥旾finished it_____. A. just ; just now B. just now ; just now C. just ; just D. just now ; just 锛?)2. 鈥昣_________ have you lived here? 鈥昐ince last year. A. How long B. How often C. How soon D. How far 锛?)3. He has been away______________. A. since a month B. for a month C. for a month ago D. in a month 锛?)4. When did your father_________________ your mother? A. marry B. marry to C. marry with D. get married 锛?)5 It _____ four years since we ______to learn English. A. Is , begin B. has been , began C. has, been D. had been ,began 锛?)6. My brother鈥檚never been late for work, __________________? A. is he B. isn鈥檛he C. has he D. hasn鈥檛he 锛?)7. 鈥旹xcuse me, when ______you ______ the man in blue? 鈥昄ast year. And I _____ him for one year. A. have ; known; have known B. did; know; knew C. did; know; have known D. have; known; knew 锛?)8. We spend as much time as wecan_____________English. A. read B. to read C. reading D. have read 锛?)9. There_______great changes in our hometown in the past fifteen years A. has been B. have been C. was D. were 锛?)10. 鈥?Hurry up. The bus is coming. 鈥?Oh, no. We mustn 鈥檛cross the street ________ the traffic light are green. A. after B. while C. since D. until 锛?)11. Italy is famous for______________ _. A. the Eiffel Tower B. the Pyramids C .the Leaning Tower of Pisa D.t he Thames River 锛?)12. 鈥?Can I see the headmaster at the moment, please?鈥旾鈥檓afraid not. He ______________out. He ___________in 10 minutes. A. goes; comes B. gone; came C. will go; will come D. has gone; will come back ( )13. He __________be at home锛宐ecause the light in his room is on. A. must B. can鈥檛C. can D. might ( )14. I think skiing is ______________great fun. A. a B. / C. an D. the( )15. 鈥?What was your brother like when he was young? 鈥?He __________ be quiet. But now he鈥檚very outgoing. A. uses B. used C. uses to D. used to 浜屻€佸畬褰㈠~绌?In the doctor鈥檚waiting room, men and women were sitting 1 on the chair, 2 for their turn. Bob, a schoolboy, was among them. They all looked very 3 except(闄や簡) Bob. He 4 an interesting story in a magazine. Just then the doctor came in to say he was ready for the next person. Bob jumped up and ran into the 5 room. 鈥淲hat鈥檚your 6 young man?鈥?said the doctor. 7 Bob said a word锛宼he doctor made him_____8_____down on a bed. 鈥淣ow锛宭et me listen to your heart.鈥?Bob tried to speak, but the doctor told him not to say anything. Bob tried to sit up, but the doctor told him 9 anything. 鈥淚鈥檒l take your temperature.鈥?Bob tried to sit up, but the doctor stopped him. 鈥淣ow open your mouth m, good.鈥?10 a moment, the doctor said, 鈥淲ell, my boy, you haven鈥檛got a bad cold. It's nothing. Mm, In fact, there鈥檚nothing wrong with you.鈥?I know there isn鈥檛锛屸€?said Bob锛屸€淚just came here to fetch(鍙栧緱锛塵edicine for my father.鈥?( )1. A. quickly B. quietly C. luckily D. noisily ( )2. A. waiting B. waited C. to wait D. wait ( )3. A. happy B. exciting C. interested D. sad ( )4. A. saw B. was seeing C. read D. was reading ( )5. A. patients B. waiting C. sitting D. doctor鈥檚( )6. A. story B. trouble C. matter D. wrong ( )7. A. Before B. After C. Until D. While ( )8. A. to lie B. lay C. lying D. lie ( )9. A. say B. not to say C. not say D. to say ( )10 A. After B. In C. On D. At 涓夈€侀槄璇荤悊瑙?A Dear Mum, Today I鈥檓going to tell you something really special. I know you work very hard and feel tired every day. Every time you come home and see the messy (鏉備贡鐨? house, I know you are very angry with me. But you always say nothing and clean up the house. I always want to help you, but I have so much homework to do. After I finish my homework, I feel like dying. I really want to help you, but I鈥檓so tired too. I also want to tell you one thing鈥旾love you so much. I usually talk back to you when you say that I don鈥檛study hard enough. But in fact I just want you to praise (璧炴壃) me more. I will try my best toget good grades and get praise from my teacher. So don鈥檛feel frustrated (澶辨湜) about my grades, OK? I have made up my mind that I will study harder and help you to clean the house. I am serious this time. I will have my homework done faster so that I can have more energy to clean the house. I will watch TV less so that I can have more time to study and get good grades. All these things are for one reason鈥旾want to make you a happy mum. I love you! Yours, Sally ( )1. Why is Sally鈥檚mother angry with Sally when she comes back from work? A. Because Sally doesn鈥檛study hard. B. Because Sally isn鈥檛good at English. C. Because Sally doesn鈥檛clean the house. D.Because Sally doesn鈥檛finish her homework. ( )2. What does Sally鈥檚mother think about her? A. She thinks Sally is very lazy. B. She feels frustrated about her. C. She thinks Sally studies very hard. D. She thinks Sally is a bad girl. ( )3. What does Sally want? A. She wants enough money. B. She wants enough free time. C. She wants good grades and praise. D. She wants enough good food. ( )4. Why does Sally want to do her homework faster? A. Because she will have enough time to watch TV. B. Because she is interested in doing homework. C. Because she has many important things to do. D. Because she can have more energy to do some cleaning in her house. ( )5. Which of the following is WRONG? A. Sally feels very tired after she finishes her homework. B. Sally loves her mother deeply. C. Sally鈥檚teacher feels frustrated about her. D. Sally wants to make her mother happy. B Diaoyu Dao and its affiliated (闄勫睘) islands, which consist of(鐢扁€︾粍鎴?Diaoyu Dao, Huangwei Yu, Chiwei Yu, Nanxiao Dao, Beixiao Dao, Nan Yu, Bei Yu, Fei Yu and other islands, are in the northeast of China鈥檚Taiwan Island, in the waters between 123掳20鈥?124掳40鈥橢(涓滅粡) and 25掳40鈥?26掳00鈥橬(鍖楃含). They belong to the Taiwan Island. The total area of these islands is about 5.69 square kilometers. Diaoyu Dao, in the western tip of the area, covers an area of about 3.91 square kilometers and is the largest island in the area. The highest part on the island stands 362 meters above the sea level. Huangwei Yu, which isabout 27 kilometers to the northeast of Diaoyu Dao, is the second largest island in the area, with a total area of about 0.91 square kilometers and a highest height of 117 meters. Chiwei Yu, about 110 kilometers to the northeast of Diaoyu Dao, is the easternmost island in the area. It covers an area of about 0.065 square kilometers and stands 75 meters above the sea level. Ancient ancestors (杩滃彜绁栧厛) in China first discovered and named Diaoyu Dao through their fishing activities on the sea. In China鈥檚historical books, Diaoyu Dao is also called Diaoyu Yu or Diaoyu Tai. The earliest historical record of the names of Diaoyu Dao, Chiwei Yu and other places can be found in the book Voyage with a Tail Wind published in 1403. It shows that China had already disc overed and named Diaoyu Dao by the 14th and 15th centuries. These historical reports clearly show that Diaoyu Dao and Chiwei Yu belong to China. The sea waters around Diaoyu Dao is traditionally锛堜紶缁燂級Chinese fishing ground. Chinese fishermen have been involved in fishing activities in these waters for generations(涓栦唬). ( ) 6. Where is Diaoyu Dao? A. In the northeast of China. B. 25掳40鈥?26掳00鈥橢. C. In the northwest of Taiwan Island. D. 123掳20鈥?124掳40鈥橢. ( ) 7. The largest island of Diaoyu Dao is ___________ square kilometers. A.5.69 B. 3.91 C. 0.91 D. 0.065 ( ) 8. The largest island of Diaoyu Dao is _______ meters taller than the second largest one. A. 75 meters B. 117 meters C. 245 meters D. 362 meters ( ) 9. How did ancient Chinese people find Diaoyu Island? A. They grew up there. B. They wrote historical books there. C. They worked and fished on the sea there. D. They drank water there. ( ) 10. Which of the following is NOT true? A. Diaoyu Dao has more than 8 islands. B. Huangwei Yu is about 110 kilometers to the northeast of Diaoyu Dao. C. China had already named Diaoyu Dao by the 14th and15th centuries. D. The sea waters around Diaoyu Dao is traditionally Chinese fishing ground. 1. The little girl will travel____________ (鍦ㄥ浗澶?to learn more about art. 2. ---Do you know Hong Kong? ---Yes锛宨t ___________ (鍥炲綊)to China in 1997. 3. There will be a__________ (榄旀湳) show at the gate of the shopping mall. 4. How to protect the________________锛堢幆澧冿級has been under discussion. 5. We should encourage ______________ not to dump waste again. Let鈥檚write a letter to them.锛堝伐鍘傦級6. Everyone was attracted by the natural ______________锛坆eautiful) . 7. Our English teacher can make her class _____________(excite) 8. These handbags are their ____________(wife) 9. He has gone to Shanghai on___________(busy) 10. The government has __________(take) action to improve the environment. 浜斻€佸畬鎴愬彞瀛?1.. I ___________________________when Mum left. 2. 鍘诲勾浠栦滑璁″垝. They ___________________________________last year. 3. 鐢靛奖?The film ___________________________ for more than an hour. 4. ? I鈥檓sure _________________________________here. 5. 褰撲粬鏀跺埌鎴戠殑淇℃椂锛岃偗瀹氫細寰堝紑蹇冪殑. He _________________________when he hears from me 鍏 1. He likes playing tricks on others, but__________, he is a good boy. He is willing to help others. 2. What's ___________the cage? 3. 卢鈥昗ill you please show me how _____________ the clock?鈥昇o problem. 4. Lots of travelling dads and moms are now using e-mail________________. 5. If you want to improve your English, practice speaking English _________. 6. 鈥?Our city is becoming more and more beautiful. 鈥?Yes, great changes ___________________ since 2006. 7. Our sitting room is____________________ Uncle Li鈥檚.8. 鈥昅um, can I have some candies? 鈥?Ok锛宐ut only one _______________. 9. There was little time left, but they ___________________ arrive here in time. 10. Tom ________________ after the whole day鈥檚hard work. 涓冦€佷换鍔″瀷闃?0鍒嗭級If you think you are too shy and want to be a little bit braver, justtry the following things: Be open to others. Tell people you are shy. There is no needto hide it. When they get to know you are a shy kid, they will understand you better. This also helps you feel more comfortable in talks. Try to smile more. When you smile, people think you are friendly and easy to talk with. Remember that other people have feelings too and most people will stay away from an angry-looking face. Learnto be a good talker. If you find it hard to start a conversation, say something nice about people around you. Think about how great you feel when someone says something nice to you. Doesn鈥檛it make you want to keep talking to that person? Get your attention elsewhere. Think more about ways to enjoy parties or games. Don 鈥檛waste time worrying about your look or whether people like you or not. You will become relaxed and find it鈥檚not so hard to talk with others. Take one small step at a time . Each time when you say 鈥淗i鈥?or smile at someone, say to yourself 鈥?You can make it.鈥?Keeping trying and one day you鈥檒l never feel shy when you talk to others.Title: Advice on 1 to be a little braverAdvice Reasons Don鈥檛2 the fact . If you tell people you are shy, they will understand you 3 Smile more. People don鈥檛4 the one with an angry-looking face. Start a conversation with 5 words. People will feel 6 and want to talk to you. Pay more attention to ways to enjoy 7 Or games. You will become 8 . Encourage yourself to say 鈥淗i鈥?or 9 at someone. Keep doing this and you鈥檒l be braver to talk to others 10 the future.?涔犺繎骞虫€讳功璁板湪鍗佷簩灞婂叏鍥戒汉澶т竴娆′?Tom 鍋ュ悍鐨勯ギ椋?紝鏈夊己鍋ョ殑浣撻瓌鏉モ€?Kate?Mary?Millie 鏈夎冻澶熺殑鏃堕棿鐜╄鈥?浣犺嚜宸?鈥?瑕佹眰锛?. 瑕佺偣榻愬叏锛屽彲閫傚綋澧炲姞缁嗚妭浣垮彞瀛?2.浜?鐐广€?3. 璇嶆暟80昏瘝鏁般€?My dream Everyone may have his own dream. Someone may want to be rich, someone may want to be beautiful, and someone may want to have power. Last Monday, we had a warm discussion about this topic at the class meeting. Here are different opinions._____________________________________________________________________ _____________________________________________________________________ _____________________________________________________________________ ________ When thinking of these things, we realize that we are full of life and hope.。
The herb medicine formula “Yang Wei Kang Liu ”improves the survival of late stage gastric cancer patients and induces the apoptosis of human gastric cancer cell line through Fas/Fas ligand and Bax/Bcl-2pathways ☆Jie Li ⁎,Gui-Zhi Sun,Hong-Sheng Lin,Ying-Xia Pei,Xin Qi,Cheng An,Jun Yu,Bao-Jin HuaDepartment of Oncology,Guang An Men Hospital,China Academy of Chinese Medical Sciences,Beijing,100053,PR ChinaReceived 19February 2008;received in revised form 8April 2008;accepted 14April 2008AbstractThe herb medicine formula “Yang Wei Kang Liu ”(YWKLF)has been used to inhibit the metastasis of human gastric cancer to prolong patient survival.In this study,we evaluated the effect of combination of chemotherapy with YWKLF on the survival of stage IV gastric cancer patients and the potential mechanisms of YWKLF by focusing on its capacity to activate apoptotic pathways in human gastric cancer cell line MGC-803.We found that combination of chemotherapy with oral administration of YWKLF significantly increased the survival of stage IV gastric cancer patients.In an approach of “serum pharmacology ”in which sera were collected from rabbits orally administered with YWKLF and examined for their anti-tumor cell activity in vitro,we observed that sera from rabbits administered with YWKLF induced the apoptosis of MGC-803cells by causing the loss of mitochondrial membrane potential,increasing the expression of Fas protein and Bax mRNA,as well as down-regulating Fas-L mRNA.Our results suggest that activation of major pro-apoptotic pathways may account for the anti-gastric cancer activity of YWKLF ,which may provide a basis for isolation and identification of more highly effective anti-cancer components.©2008Elsevier B.V .All rights reserved.KEYWORDSHerb medicine;Serum pharmacology;Apoptosis;Gastric carcinoma;Bax/Bcl-2;Fas/Fas ligandAbbreviations :YWKLF ,formula “Yang Wei Kang Liu ”;Sub-F1,Sub-Formula 1;Sub-F2,Sub-Formula 2;Sub-F3,Sub-Formula 3;AgNORs,argyrophilic nucleolar organizer regions;AO,acridine orange;Δψm ,mitochondrial membrane potential.☆This project was supported by grants No.30600839and No.30672764from National Natural Science Foundation of China.⁎Corresponding author .Tel./fax:+861088001340.E-mail addresses:drjieli2007@ ,drjieli@ (J.Li).1567-5769/$-see front matter ©2008Elsevier B.V .All rights reserved.doi:10.1016/j.intimp.2008.04.007w w w.e l s ev i e r.c o m /l o c a t e /i n t i m pInternational Immunopharmacology (2008)8,1196–12061.IntroductionGastric cancer is one of the most common malignant tumors in China [1].Facing the problems of lower rate of early diagnosis,higher possibility of relapse and metastasis after surgery,and poor sensitivity to radiotherapy or/and che-motherapy,herb medicine plays a role in preventing gastric cancer from relapse and metastasis [2].A herb medicine formula “Yang Wei Kang Liu ”(YWKLF),consisting of six herbs,has been used for a number of years in traditional medicine to promote circulation and human body resistance to pathogens and cancer .Preliminary clinical studies have shown that YWKLF modulates immune function,improves the quality of life,and prolongs survival time of cancer patients [3].However ,the molecular mechanisms underlying the anti-cancer effect of YWKLF are poorly understood.Apoptosis is an important component of various biological processes,such as development,maintenance of tissue homeostasis and elimination of transformed cells.The aberrant regulation of apoptosis contributes to many types of human diseases including cancer.Apoptosis uses two main signaling mechanisms:the “extrinsic ”and the “intrinsic ”pathways,to trigger programmed death of cells.The Fas –Fas ligand (FasL)pathway is the most studied in the cells through extrinsic pathway [4–6].The Bcl-2family of proteinsconstitutes a critical intrinsic checkpoint coupled to cell apoptosis.The ratio of anti-to pro-apoptotic molecules,such as Bax/Bcl-2,sets the threshold of cell susceptibility to apoptosis of the Bax/Bcl-2pathway,which utilizes organelles such as mitochondria to amplify death signal [7].Our previous study found YWKLF regulated Fas/FasL expression on T lymphocytes of gastric cancer patients [8].Thus,it is possible that YWKLF may inhibit human cancer by inducing apoptosis.Due to the difficulties in standardizing the studies of formulated herb medicine,“serum pharmacology ”has been proposed by Japanese scholars in the 1980s [9],in which a single herb or herb mixture is administered orally to animals,followed by collecting animal sera for testing on the biological behavior of tumor cells.The methodology has been believed to better reflect the maintenance of herb components and metabolism through a host,therefore may be more informative than drug extracts when tested in vitro [10].In the present study,we report the effect of combination of YWKLF with chemotherapy on prolonging survival of stage IV gastric cancer patients.In addition,we report that sera from rabbits administered with YWKLF promote the apopto-sis of human MGC-803gastric cancer cells through Fas/Fas ligand and Bax/Bcl-2pathways.2.Materials and methods2.1.Patients and the treatmentA total of 123patients with stage IV gastric cancer were enrolled during the period of 1990to 1997.The study was permitted by Institutional Review Board (IRB),and informed consents (IC)were obtained from the patients before they were recruited into this study.All patients had Karnofsky performance status N 60and showed adequate hematological,liver and renal functions.Patients were recruited to YMKLF +chemotherapy and chemotherapy alone cohorts by random number tables with informed consent.Patients assigned to chemotherapy group received as MFP chemotherapy (cisplatin 60mg/m 2on day 1,5-FU 500mg/m 2/day,on day 1–5and MMC 6mg/m 2/day,on day 1,8)every 4weeks,for 4cycles.In chemotherapy +YWKLF group:patients received MFP chemotherapy in combination with oral administration of lyophilized YWKLF granules,12g twice daily.Changes in tumor size and Karnofsky performance status were analysed analyzed every 2months.2.2.Cells and reagentsHuman gastric cancer cell line MGC-803was purchased from Cell Culture Centre,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences and cultured in RPMI 1640(GIBCO/BRL)containing 10%fetal bovine serum (FBS).Rabbit anti-human Fas and FasL polyclonal antibodies and FITC-labeled goat anti-rabbit IgG antibody were obtained from Santa Cruz Corporation (Beijing,China).Annexin V-FITC Apoptosis Detection Kit was from BD Corporation (Shenzhen,China).TRIzol Reagent was purchased from Invitrogen/GIBCO (Beijing,China).Reverse Transcriptase,Taq DNA polymerase and other reagents used in reverse transcription PCR (RT-PCR)were obtained from Promega Corporation (Beijing,China).2.3.AnimalsJapanese flap-eared rabbits were purchased from the Institute of Laboratory Animal Science,Chinese Academy of Medical Sciences (CAMS)(Beijing,China)and maintained in the animal facility approved by institutional policies.2.4.Formula preparationYWKLF is composed of Astragalus mongholicus Bge (450g),Panax notoginseng (Burk.)F .H.Chen (40g),Panax ginseng C.A.Mey (100g),Paris polyphylla Smith var .chinensis (Franch.)Hare (150g),Caesalpinia sappan L (60g)and Oldenlandia diffusa (Willd.)Roxb (180g).Based on the principles of herb action,YWKLF was sub-formulated into Sub-Formula1(Sub-F1):Astragalus mongholicus Bge (450g),Panax ginseng C.A.Mey (100g),that improve the general body conditions [11];Sub-Formula 2(Sub-F2):Panax notoginseng (Burk.)F .H.Chen (40g)and Caesalpinia sappan L (60g),that improve circulation [12–14];and Sub-Formula 3(Sub-F3):Paris polyphylla Smith var .chinensis (Franch.)Hare (150g)and Oldenlandia diffusa (Willd.)Roxb (180g),that contain anti-oxidants [15].All medicinal plants used to prepare formulae were provided by Guang An Pharmacy (Beijing,China),the species,plant parts,and origin used in the formula as Table 1.The plant were homogenized with a Waring blender ,then soaked in 12Liter double distilled water (DDW)for 24h.The mixture was heated to 100°C for 1h,and the decoction was filtrated.The filtrates obtained from 3cycles of the procedures were mixed,concentrated by heating and granulated by lyophilization.Total1197YWKLF prolonged the overall survival time of stage late gastric cancer patientsyield of the YWKLF extract is 125g lyophilized powder from water extract of 1kg raw mixed herb.Aqueous solution was prepared by dissolving the granulated formulae in sterile water for animal administration at 1.63g raw mixed herb/ml for YWKLF ,0.92g raw mixed herb/ml for Sub-F1,0.17g raw mixed herb/ml for Sub-F2and 0.55g raw mixed herb/ml for Sub-F3.The quality control on YWKLF preparation,including definition of the correct plants,origin of production,implantation,harvesting,and processing.Good manufacturing practice has been used to produce lyophilized particles.Contamination of hazardous agents such as pesticide residues,arsenic and heavy metals,microorganisms including aflatoxins,and sulfur dioxide residue were excluded.Consequently,YWKLF and its preparations were standardized,regulated and quality controlled according to the guidelines defined by Chinese State Food and Drug Administration (SFDA).2.5.Method of “serum pharmacology ”“Serum pharmacology ”has been used to study the in vitro pharmacological activity of herb medicine.The preparation of “serum pharmacology ”was based on the described procedures [16].Briefly,18rabbits were randomly divided into six groups and administered by gavage of YWKLF ,Sub-F1,Sub-F2and Sub-F3.The rabbits of saline and chemotherapy groups were fed with 10ml saline twice daily for 3days per rabbit.YWKLF and its sub-formula groups were treated with 10ml YWKLF or a sub-formula twice daily for 3days per rabbit.The formula dose was estimated as 10times of doses given to patients in the clinic.T wo hours after the fifth treatment on the third day,blood was obtained from aorta of the rabbits under sterile conditions,and was allowed to coagulate at 25°C for 4h.The sera were separated by centrifugation at 2500rpm for 20min.After filtration through 0.45μm cellulose acetate membrane twice,the sera were treated in 56°C water bath for 30min then were stored at −20°C until use.2.6.Cell viability assay by MTTCell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)dye reduction assay (Sigma,USA).MGC-803cells were seeded at a density of 1×104cells per well into 96-well plates in RPMI 1640containing 10%FBS.After 24h,the cells were washed and further cultured in the following media:the saline groups:RPMI 1640containing 10%,20%and 30%sera from rabbits treated with saline;the YWKLF groups:RPMI 1640containing 10%,20%and 30%sera from rabbits treated with herb formulae;the chemotherapy groups:10%,20%and 30%sera from saline saline-treated rabbits in RPMI 1640plus cisplatin at a final concentration of 2μg/ml.The cells were incubated in 5%CO 2at 37°C for 48h,then with 5mg/ml MTTat 10μl/well for 4h.The culture media were discarded and substituted with DMSO at 100μl/well.The plates were mounted on a micro-mixer for 5min to thoroughly dissolve the blue MTT/DNA granules.The plates were then placed in a microplate reader .The OD was measured at 570nm.The inhibition rate was calculated according to the following formula:Inhibition rate k ðÞ¼100k Â1Àmean absorbance of cells in sera from drug treated rabbits =mean absorbance for cells in sera from rabbits treated with saline ðÞf g :2.7.Morphometrical study of argyrophilic protein associated with the nucleolar organizer regions (AgNORs)in MGC-803cellsAfter 24h treatment with rabbit sera,200μl cell suspensions was loaded into a cytospin chamber and spun for 10min at 300rpm.Slides were fixed by a mixture of methanol and glacial acetic acid at 3:1vol/vol.The silver colloid solution for AgNORs staining was prepared by dissolving gelatin in 1%aqueous formic acid to a concentration of 2%,and this solution was mixed at 1:2vol/vol with 50%aqueous silver nitrate in which slides were immersed for 45min at 20°C in dark [17,18].After rinsing in H 2O,the slides were immersed in 5%sodium thiosulfate for 45min at room temperature.Counterstaining was carried out using methyl green.The parameters of nucleolin and AgNOR were assessed by an image analysis system (KL-2,Beijing).The stained dots of only one focal plane per nucleus,scanned from light microscopy image using a ×100oil-immersion lens,were converted via a TV camera to appear as dots on a high-resolution monitor .The mean number and area of stained dots were measured using an image analyzer .Measurements were made for 150cells in random areas of each specimen.The morphometric parameters were means of areas and numbers of AgNORs per nucleus and the ratio of AgNORs area to nucleolin/nucleus area per cell.2.8.Detection of cell apoptosis by flow cytometryMGC-803cells seeded in 6-well plates at 106/well were cultured in the presence of sera from rabbits received saline or formulae for 24h.The cells were then washed and fixed in 70%ethanol at 4°C overnight.The cells were then re-suspended in 10μl propidium iodide (PI)solutionTable 1The component of YWKLFSpecies (family)Plant part OriginSub-Formula 1Astragalus mongholicus Bge Root Neimenggu,China Panax ginseng C.A.Mey Root JiLin,China Sub-Formula 2Panax notoginseng (Burk.)F .H.Chen RootYunnan,China Caesalpinia sappan L Heartwood Yunnan,China Sub-Formula 3Oldenlandia diffusa (Willd.)RoxbLeaves Henan,China Paris polyphylla Smith var .chinensis (Franch.)HareRhizomeYunnan,China1198J.Li et al.(2mg/ml)with 50μl RNAase (10mg/ml)and incubated in the dark at 37°C for 30min,followed by DNA content analysis using a FACSort flow cytometer (Becton Dickinson).The CellQuest program was used to analyze the results.Different phases of cell cycle were assessed by collecting the signal at channel FL2-A.The percentage of the cell population at a particular phase was estimated by ModFit LT .Annexin V-FITC/PI staining was performed after the cells were treated with rabbit sera for 24h in vitro.Rabbit sera treated cells (2×106/group)were rinsed with ice-cold PBS and then re-suspended in 200μl of reaction buffer .Annexin V stock solution (10μl)was added to the cells and incubated for 30min at 4°C.The cells were further mixed with 5μl PI and immediately analyzed on a FACSort equipped with CellQuest software.2.9.Assessment of morphological changes by acridine orange (AO)stainingCell slides were fixed in 95%ethanol for 10min on ice and air dried for 10min.The slides were stained with AO (6μg/ml;sigma),then examined by laser scanning confocal microscopy (Radiance2000,BIO-RAD)using a 488nm argon laser excitation beam,a 515nm beam filter for DNA,and a 630nm beam filter for RNA.Apoptotic cells were characterized by morphological alterations such as condensed nuclei and cell shrinkage.AO is an intercalating,nucleic acid specific fluorochrome,which emits green fluorescence when bound to DNA.Under fluorescence microscopy,viable cells show bright green nuclei with intact structure while apoptotic cells exhibit bright green nuclei with condensed chromatin in dense green areas.Three to five randomly chosen fields were viewed in each slide with a minimum of 300cells for each group.2.10.Analysis of mitochondrial membrane potentialMitochondrial membrane potential (Δψm )was assessed by rhodamine 123,a mitochondrial potential sensor (Molecular Probes).Cells were seeded in six-well plates for 6h.The medium of each well was discarded and treated with 1ml of medium containing rhodamine 123at 25μM for 30min at 37°C and 5%CO 2in the dark.The cells were washed three times in PBS,then were analysed analyzed by flow cytometry.2.11.Fas and FasL detection by flow cytometrySera-treated cells washed twice in PBS buffer were incubated with a rabbit anti-human Fas antibody or a rabbit anti-human FasL antibody at 4°C for 30min.FITC-labeled goat anti-rabbit IgG antibody was then added to cells for 30min at 4°C.Flow cytometric analysis was performed using a Becton Dickenson FACSort and CellQuest software.A total of 10,000events were acquired for each analysis.2.12.Detection of FasL,Bax,Bcl-2mRNA in MGC-803cells by RT-PCRAfter rabbit sera treatment for 24h,MGC-803cell RNA was extracted with TRIzol Reagent,and used for reverse transcription RT-PCR.For PCR amplification,specific primers used were as follows:FasL (299bp):5′-GGAGCTGGCAGAACTCCGAGAG-3′and 5′GGCTCAGGGGCAGGTTGTTG-3′;Bax (312bp):5′-GCGTCCACCAAGAAGCTGA-3′and 5′-ACCACCCTGGTCTTGGATCC-3′;Bcl-2(231bp):5′-CAGCTGCACCTGACGCCCTT-3′and 5′-GCCTCCGTTATCCTGGATCC-3′;β-actin primers:5′-CGTCTGGACCTGGCTGGCCGGGACC-3′and 5′-CTAGA AGCATTTGCGGTGGACGATG-3′(600bp).The PCR conditions for FasL were:denaturation at 94°C for 50s,annealing at 63°C for 50s,and extension at 72°C for 1min,40cycles.For Bcl-2and Bax:denaturation at 94°C for 50s,annealing at 62°C for 50s,and extension at 72°C for 1min,35cycles.The PCR products were separated by electrophoresis on 2%agarose gels which were stained with ethidium bromide.2.13.Statistical analysisAll in vitro experiments were performed for at least 3times and the mean ±SD of representative results are presented.Significant differences among the groups were determined using two way analysis of parison of the rate of the patient survival was analyzed by Kaplan –Meier survival analysis method.P b 0.05was considered as an indication of statistical significance.Table 2Clinical characteristics of patients with stage IV gastric cancerChemotherapyChemotherapy +YWKLF Total no.of patients 4677Sex:male/female 35/1159/18Histological type Adenocarcinoma3253Mucinous adenocarcinoma 1220Adenosquamous carcinoma 24Disease statusRelapse after surgery 820Bypass surgery 1421Laparotomy 1217No surgery12191199YWKLF prolonged the overall survival time of stage late gastric cancer patients3.Results3.1.The effect of combination therapy of YWKLF with chemotherapy on the survival of stage IV gastric cancer patients77patients were assigned to treatment with chemotherapy plus YWKLF and 46patients were assigned to chemotherapy alone.The background factors of patients as Table 2.There is no statistically significant difference of the background in the patients between chemotherapy alone and chemotherapy with YWKLF cohorts in terms of age,sex and tumor histology.The rate of response (RR)as reduction by at least 50%in the size of primary tumor in 46patients receiving chemotherapy alone was 13.04%(6/46),and 33.77%(26/77)of patients receiving chemotherapy and YWKLF showed responses.The median survival time was 14months for chemotherapy alone and 21.5months for YWKLF +chemotherapy patients (P b 0.001)(Fig.1).These results indicate the advantage of combination of chemotherapy with YWKLF in prolonging the survival of stage IV gastric cancer patients over chemotherapy alone.3.2.The effect of YWKLF rabbit serum on MGC-803cell viabilityWe next examined the effect of YWKLF on the biological behavior of human gastric cancer cells.Pervious studies of serum pharmacology suggested an eight to eighteen times of drug doses should be needed to administer into rabbits as compared to humans.Based on our clinical data and preliminary experiments,we chose a rabbit administration dose that equaled to 10times of doses given to patients.Administration with herbal mixtures did not cause the loss of body weight,decreased activity,and change in fur quality of the rabbits (data not shown).By using “serum pharmacology ”,we harvest sera from rabbits administered with YWKLF and sub-formulae andtested their effects on the growth of the MGC-803human gastric cells in vitro.As shown in Fig.2A,after treatment for 48h,with serum from rabbits received YWKLF ,MGC-803cells showed significantly reduced rate of growth.The maximal cell growth inhibition was observed when serum from YWKLF rabbits was used at 20%concentration in the cell culture with slightly decreased effect at 30%concentration reflecting “plateaued ”biological responses seen in many drug experiments.The results also suggest that serum harvested 2h after last gavage administration of YWKLF was optimal in inhibiting cell growth.When sera from rabbits administered with sub-formulae were tested,Sub-F1and Sub-F3sera showed significant inhibition of cell growth,although the efficacy was lower than the intact formula serum.In contrast,Sub-F2serum showed little inhibitory activity on cell growth.Thus,components in both Sub-F1and F3contributed to the overall activity of YWKLF in “serum pharmacology ”test.It should be noted that YWKLF and its sub-formulae did not affect the viability of normal gastric cells in primary culture or endothelial cells (data not shown).3.3.YWKLF serum inhibited AgNORs activity in MGC-803cellsThe levels of argyrophilic nucleolar organizer regions (AgNORs)represent cell cycle kinetics and the shorter the cell cycle,the greater rate of the synthesis of rRNA,therefore,the increased quantity of Ag NORs present in the nucleoli.Thus,AgNOR value was used to measure the rate of cell proliferation.PreviousFigure 2Inhibition of MGC-803cell proliferation in vitro by sera from formula-treated rabbits.MGC-803cells were treated with sera from rabbits administered with YWKLF ,Sub-F1,Sub-F2,and Sub-F3for 48h.Cell growth was determined by MTT .A.The effect of rabbit sera at different concentrations in cell culture. B.Inhibition of cell growth in the presence of sera (20%),1–2h after the last formula administration in rabbits.⁎⁎P b 0.01,compared with the effect of serum from rabbits given saline (0%inhibition).Figure 1Survival of stage IV gastric cancer patients with chemotherapy alone or in combination with YWKLF .Kaplan –Meier analysis of survival of patients treated with chemotherapy or chemotherapy+YWKLF .The median survival time was 14months for chemotherapy group and 21.5months for chemotherapy+YWKLF group,respectively .P b 0.001,significantly increased survival as compared with patients receiving chemotherapy alone.1200J.Li et al.studies showed a linear increase in both the area and the number of AgNOR stainings in each tumor cell with increasing degree of malignancy[18].We found AgNORs as distinct black–brown dots on a yellow pale background(Fig.3A),and the black–brown dots in the YWKLF serum treatment cell group were smaller than cells treated with serum from saline administered rabbits(Fig.3B). The ratio of AgNORs area to nucleolin/nucleus area per cell(I/S) is significantly decreased in cells treated by YWKLF serum and saline serum containing cisplatin(⁎P b0.05).Sub-F2and Sub-F3 sera did not affect I/S in MGC-803cells,while Sub-F1serum decreased I/S in MGC-803cells,but with the efficacy lower than YWKLF serum(Fig.3C).Thus,YWKLF rabbit serum inhibited MGC-803cell proliferation as shown by reduced AgNORs activity in the nuclei.3.4.YWKLF rabbit serum induced MGC-803cell apoptosis We evaluated sub-diploid DNA content(sub-G1)in MGC-803cells using flow cytometry.As shown in Fig.4A,YWKLF rabbit serum treatment resulted in an increase in the sub-G1phase of MGC-803 cells after24h(19.58±2.1%versus2.66±1.2%cells with saline rabbit serum).We did not observe significant effect on sub-G1 peak by Sub-F2and Sub-F3sera(Fig.4A).However the sub-G1 peak in cells treated by Sub-F1rabbit serum was12.36±2.3%, lower than YWKLF serum,but the effect was statistically significant.The number of cells in the G2/M phase was significantly decreased after treatment with YWKLF rabbit serum for24h with an increase in S-phase cell number.These results indicate MGC-803cell cycle arrest at S phase after treatment with YWKLF rabbit serum(Fig.4B).The chemotherapy agent cisplatin has been reported to increased the fraction of S-phase cells in prostate cancer cells[19]and induced S-phase arrest of human ovarian carcinoma cells[20].In this study,Fig.4A shows cisplatin(2μg/ml)in saline rabbit serum induced S-phase arrest of MGC-803cells after24h treatment,with an increase in cells in apoptotic sub-G1phase(25.86±3.1%).These results that indicate YWKLF rabbit serum was able to prevent gastric cancer cell division.We then evaluated MGC-803cell apoptosis by using annexin V-FITC and PI staining.Fig.4C showed that MGC-803cells treated with saline rabbit serum remained Annexin V−/PI−. YWKLF rabbit serum treated cells increased Annexin V+/PI−staining.In contrast,cispaltin in saline rabbit serum induced Annexin V+/PI+cells.Cisplatin caused both the early and late stage apoptosis in gastric cancer cells,while YWKLF rabbit serum preferentially induced early stage cancer cell apoptosis.Sub-F2 and Sub-F3rabbit sera did not increase Annexin V+/PI−and Annexin V+/PI+cells(Fig.4C).We further determined morphological changes in MGC-803 cells by confocal scanning laser microscopy.Fig.5A-1showsthe Figure3Argyrophilic nucleolar organizer regions(AgNORs)in MGC-803cells.A.Cells cultured in the presence of20%serum from rabbits given saline.B.Cells cultured in the presence of20%serum from rabbits given YWKLF.X axis indicates cell numbers,Y axis defines I/S%.C.The ratio of AgNORs area to nucleolin/nucleus area per cell.A total of150random cells were counted under a light microscope(×400).⁎P b0.05compared with cells cultured with serum from rabbits given saline.1201 YWKLF prolonged the overall survival time of stage late gastric cancer patientschange in gastric cancer cell morphology after exposure to saline-treated rabbit serum,which failed to induce apoptosis.Mean-while,MGC-803cells showed features of apoptosis,including cell shrinkage,nuclear condensation,DNA fragmentation,break down of the cells and formation of apoptotic bodies,after 24h exposure to YWKLF rabbit serum and Sub-F1rabbit serum (Fig.5B).After YWKLF rabbit serum treatment,the nuclear structure of MGC-803cells became incomplete (Fig.5C),supporting the notion that YWKLF rabbit serum induced cell apoptosis.3.5.YWKLF rabbit serum induced mitochondrial dysfunction in tumor cellsMitochondria play a critical role in apoptosis induced by cancer therapeutic agents [21,22],which cause mitochondrial mem-brane potential (Δψm )loss and cytochrome c release into the cytosol,inducing caspase-9-dependent activation of caspase-3and cleavage of DNA reparatory protein PARP .We examined the effect of YWKLF rabbit serum on Δψm using rhodamine 123,a mitochondrial potential sensor .A marked loss of Δψm was observed after exposure of MGC-803cells to YWKLF rabbit serum for 4to 24h.The Sub-F2and Sub-F3rabbit sera did not affect cell Δψm ,whereas,Sub-F1rabbit serum caused Δψm loss in cancer cells at the levels similar to YWKLF rabbit serum (Fig.6).3.6.Fas/FasL expression in MGC-803cellsThe Fas –FasL interaction is a key physiological regulator of programmed cell death [23].We investigated the expression of Fas/FasL in MGC-803cells by flow cytometry and foundthatFigure 4Induction of MGC-803cell apoptosis by sera from rabbits given formulae.A.Flow cytometry analysis of cell cycle distribution of cultured MGC-803cells with PI staining following 24h exposure of cells to rabbit sera.Arrows indicate the population of cells with sub-G1phase DNA content,suggestive of apoptotic cell death.B.Distribution of MGC-803cell cycle indicated by PI staining.The values shown are the mean ±SEM of 3experiments.⁎⁎P b 0.01compared with the effect of serum from rabbits given saline.C.Detection of apoptosis with Annexin V-FITC/PI binding to MGC-803cells.Viable cells (Annexin V −/PI −)are located in the lower left,apoptotic cells (Annexin V +/PI −)in the lower right,post apoptotic secondary necrotic cells (Annexin V +/PI +)in the upper right and primary necrotic cells (Annexin V −/PI +)in the upper left quadrants,respectively.Numbers in each quadrant are percentage of cells.1202J.Li et al.。