歌词中的高潮部分to be some plaing now的女声版的英文歌叫什么？

Here to stay
威名依旧
Even when I'm gone
即便我已离开
When I close my eyes
当我闭上双眼
Through the passage of time
穿过时间的洪流
Kings never die
王者永生
I can hear the drummers drumming
我可以听到鼓声隆隆
And the trumpets
小号齐鸣
Someone's tryna summon someone
有人在召集众人
I know something's coming
我知道审判日即将到来
But I'm running from it to be standing at the summit
但我远远逃离，只为伫立顶峰
And plummet
然后猛冲直下
How come it wasn't what I thought it was
一切并非如我所想那般发生
Was it, too good to be true
那是否太过美好，而显得并不真实？
Have nothing, get it all but too much of it
从一无所有到拥有一切，是不是得到的太多了？
Then lose it again, then I swallow hallucinogens
再次一无所有 我磕药让自己释怀
'Cause if not, where the hell did it go
因为如若不然，这将向何处发展？
'Cause here I sit in Lucifer's den by the dutch oven
我坐在堕落天使的巢穴中，坐在荷兰烤箱旁
Just choosing to sin
我选择堕入罪恶
Even if it means I'm selling my soul
即使意味着我要出卖我的灵魂 我
Just to be the undisputed again
也要再次成为不容置疑的终极斗士
Do whatever I gotta do this to win
不惜一切代价只为战无不胜
'Cause I got this motherfucking cloud over my head
因为那该死的乌云蒙蔽了心
Crown around it, thorns on it
戴上皇冠，披上荆棘
Cracks in it, bet you morons didn't
冲破云霄，我打赌，你们这些愚人做不到
Think I'd be back, did ya
你们以为我会再次归来，对吗？
How 'bout that I'm somehow
那我现在再回去变成你们这些失败者
Now back to the underdog
怎样？
But no matter how loud that I bark
但是无论我像疯狗一样吠地多响
This sport is something I never bow-wow'd at
这游戏是我从来不会屈服的
I complain about the game
我抱怨这游戏规则
I shout and I pout, it's a love-hate
我大声呼喊，气愤不已，那是一场爱恨情仇的游戏
But I found out that
但是我发现
I can move a mountain of doubt
可以打消我心头那像山一样沉重的疑惑和痛苦
Even when you bitches are counting me out
即便你们这些混蛋将我驱逐
And I appear to be down for the count
让我陷入情绪低潮
Only time I ever been out
只有时间能将我治愈
And about is driving around town
常常会选择在城里飙车
With my fucking whereabouts, in a doubt 'cause
无人知晓我的踪迹
I been lost tryna think of what
因为我已迷失，想要弄明白
I did to get here but I'm not a quitter
如何能找回自我 但我不会轻言放弃
Gotta get up, give it all I got or give up
绝不会让所有努力付诸东流，就此放弃
Spit or shit or stepped on but kept going
练习说唱 短暂小息，或者进步，但是绝不停下
I'm tryna be headstrong but it feels like
我努力勇往直前 但感觉
I slept on my neck wrong
似乎坚持错了方向
'Cause you're moving onto the next
你开始往下一个目标前进
But is the respect gone
是否曾经所有崇敬已消逝？
'Cause someone told me that
但是有人曾经告诉我
(Kings never die)
王者永生
Don't give me that sob story liar
不要给我那些讲虚假的煽情故事，
Don't preach to the choir
不要找借口 更不要不要白费唇舌
You ain't ever have to reach in the fire to dig deep
你都从没直面那火焰，坚持挖掘真相
Nobody ever handed me shit in life, not even a flyer
这糟糕的生活中，无人对我施予援手
Wouldn't even take shit into consideration
我才不在乎其他一切与我无关的事
Obliterate anyone in the way
这条路上抹去所有人
I think I see why a lot of rappers get on these features
我想我知道为什么那么多rappers想要在一首歌里各种炫技
And try to show out on a track with me
想跟我合作来给他们的歌添彩
But it'd actually have to be
但是 真的很差劲的
A fucking blowout to get me to retire
表演就想让我退休
Tell these new artists that kings never die
我要告诉你们这些新起之秀，王者永生
I know shit has changed in this age
我知道，在这时代，一切都在改变
Fuck a Twitter page
去他的推特首页
Did just say I've been upstaged
你是说我大势已去了吗？
Why am I online It's driving me crazy
我为什么要在网上？这让我抓狂
I'm riding shotgun tryna get a gauge
我拿着q想要搞清楚
Almost hot, but I'm not running kaboom
我在网上挺火的，但是并没有引起大轰动，
With a stage pass and this shit
只是得到上台舞台表演的机会
And opinions sway, I can hear them say
各种言论纷纷扬扬 我可以听到他们在说
If I stay passionate maybe
如果我保持热情
I can stay Jay miraculous
那么我可以创造奇迹
Comeback as if I went away, but detractors
要我卷土重来，
Just say some bullshit for the Renegade
看起来好像我曾经离开过
Someone's gonna make me blow my composure
有些人让我无法冷静下来
Here I go again, to the stage
现在我重新开始，走上舞台
And I feel like I'm in a cage
我感觉好似身陷笼中
They so want a champion to fall
他们渴望看到冠军惨败
I still wonder why I laugh at 'em
难怪我会取笑他们，
'Cause why carry when I'm all
因为他们从来都赢不了我
So fuck what these cynics say
去他的愤世嫉俗之言
Just goes to show
即便我深陷困境
That when my back's against the wall
我也会让他们看看我的不屈
And I'm under attack again
而我现在又再次成为攻击的目标
I'll act as if I'm posted up
我也会像是众所周知般的勇敢面对
With this pented rage
这幽闭的笼子里
'Cause saw all these plaques in my office
看着录音室里白金唱片
On the floor stacked against the door
一直堆放到到门边
All be just metaphors for the odds of
难道它们也单单只是象征
When you comin' back again
为了叫我回到从前的水平
'Cause all the accomplishments, accolades, awards
那些所有的成就，荣耀，奖项
And trophies just don't mean jack anymore
其实根本就什么意义都没有
'Cause I'm here today and gone tomorrow
因为即使我今天活着，我也可能明天就会逝去
And I'm not gonna be
我不会永恒
Here to stay
威名依旧
Even when I'm gone
即便我已离开
When I close my eyes
当我闭上双眼
Through the passage of time
穿过时间的洪流
I want it, I'm coming to get it
我想拥有一切，我来此争取将其拥有
So you son of a bitches don't duck
所以，你们这些混蛋不要
Your gun and get rid of those
再叫嚣
Critics all end up in critical, think your shit
我会摆脱掉那些最终躺在急症室的你们
Is dope all you're gonna get is smoked then
你们自以为你自己很厉害 你们只能被我摧毁地尸骨无存
And I'm not stopping until I'm top again
我不会停下脚步，直到我再次登顶
All alone and on a throne
独拥宝座
Like a token of respect, or a homage poem
向我表示尊重，荣耀的诗歌
Or an ode, I've been on
或者一首颂歌，我就会永生
Tossed in the air by my own arm
将我自己投掷至空中
And launched so hard I broke my collarbone
向上发射，用力过猛我折断了我的锁骨
And it's my time to go
你们认为是我应该离开的时候
But I'm still not leaving
而我不会就此离开
Stop for no one, I don't know
没人可以阻止我，我也不清楚
But I've been told that I should go
但是我被告知
Launch a Rover, knock it over
如果有障碍挡在你面前，摧毁它
Time to go for that pot of gold, 'cause
是时候准备那个绝杀了
They say kings never die
他们说，王者永生
Just ask Jam Master Jay
尽管看看Jam Master Jay
They just grow wings and they fly
他们长出了羽翼，遨游飞翔
So, hands up, reach for the sky
所以，高高举起你们的手
Try to hold up and provoke these moments
努力铭记这辉煌时刻
'Cause in a blink of an eye
因为眨眼间
They'll be over
一切便会结束
Tryna secure your legacy like Shakur
像2PAC那般，书写自己的传奇
And ensure that nobody's
确保无人
Ever gonna be what you were
能与你的辉煌战绩相匹敌
So before you're leaving this Earth
所以，在你离开这个星球前
You want people to feel the fury
你想让人们感受到
Of a pure evil cerebral berzerk
那前所未有狂怒
Deacon of words, syllable genius at work
执事祝辞，音节天才
Plus I'm think that they're mistaking
另外，我觉得他们误解了
My kindness for weakness, distill it with meanness
我的仁慈和怯弱只是一味谈论我的刻薄、卑劣
With some kind of back flipping burgers
他们吃着烤架上一些反面
On the grill for some peanuts
带有花生汉堡包说道
Secure us to arenas, call me Gilbert Arenas
确保我们能上圆形斗兽场，叫我Gillbert Arenas
Still appeal to the dreamers
我仍旧着迷于那些追梦者
I made it to the silverscreen
我大获成功
Where Rocky stepped with the demons
跻身于Rocky曾与恶魔周旋的地方
Khalil on the beat 'cause making the beat
Khalil正在战场上，因为我从没觉得我们是在制作节奏
Ain't the same feeling to me is killing the beat
我们是在毁灭中产生节奏
It's fulfilling to me, what filling a seat is
这让我感觉到我真的是实实在在坐在王位上的感觉
That sound, vomit, thirst
旋律，嘶吼与渴望
And how common underground commons eat
以及生活在贫民区时那些救济粮
Outsider stomach growl, throbbing hunger
到处寻找机会
Out-rhyming everyone, God just give me one shot
但上帝只给我这一次成功的机会
I swear that I won't let you down
我发誓，我不会让你失望
I'mma be around forever
即便长眠于土
Entertain even in the ground
我也会永垂不朽
You ain't ever gonna hear me say I ain't
你绝对不会听到我说我并非永恒
Here to stay
威名依旧
Even when I'm gone
即便我已离开
When I close my eyes
当我闭上双眼
Through the passage of time
穿过时间的洪流
Kings never die
王者永生
Kings never die
王者永生

现在很多浏览器都提供了自动刷新标签的功能,例如傲游浏览器、360浏览器等,直接在标签页面中设置自动刷新频率即可,但是这样还需要不定时人工查看是否有更新才行。而如果目标页面有变化或达到预期要求时自动发出提醒,不妨使用一款小软件(NotiPage),通过点击按钮New aleet按钮添加一个监控任务,再设置好监控的目标页面、自动刷新的时间,以及监控类型(提供监控页面变化、特定关键字是否显示及指定网页是否启用等),软件就会自动帮助监控,一旦有变化就会d出窗口提示了。

在chrome上安装chrome插件：Page Monitor
安装好之后会在chrome的右上角出现类似心电图一样的图标。
右键该图标再点击“选项”。
任意选一个你需要监测的网页的标签，再左键点击p m,再点击“监控此页”。
此时回到p m的选项页面，页面已经出现对刚刚 *** 作的体现，那么我们继续设置下提醒的方式，检查的时间间隔。设置后会当即生效。
看到该三项设置的下面有个高级的按钮了么，点击高级。
设置更人性化的提醒方式。
设置具体项目的下方有导入和导出监测页面的选项，给我们批量设置提供了便利，点击立即检查全面会当即检查一遍所有监测的页面，如有变化将马上做提醒。

现在很多浏览器都提供了自动刷新标签的功能,例如傲游浏览器、360浏览器等,直接在标签页面中设置自动刷新频率即可,但是这样还需要不定时人工查看是否有更新才行。而如果目标页面有变化或达到预期要求时自动发出提醒,不妨使用一款小软件(notipage),通过点击按钮new
aleet按钮添加一个监控任务,再设置好监控的目标页面、自动刷新的时间,以及监控类型(提供监控页面变化、特定关键字是否显示及指定网页是否启用等),软件就会自动帮助监控,一旦有变化就会d出窗口提示了。

make （verb）
1
▶PRODUCE◀
[transitive] to produce something, for example by putting the different parts of it together
I'm going to show you how to make a box for your tools A family of mice had made their nest in the roof She made her own wedding dress The company has been making quality furniture for over 200 years They met while they were making a film for Australian TV Make a list of all the things you need
make somebody something
He made her a toy horse, using just some straw and bamboo twigs
be made from something
Paper is made from wood
be made (out) of something
a shirt made of silk
make something from/out of something
She's very good at making things from old scraps of material
Japanese-made/English-made etc
(=produced in Japan etc)
2
▶DO◀
[transitive] used with some nouns to say that someone does something
Anyone can make a mistake I can't make a decision just yet I need to make a quick phone call You could have made more effort to talk to him He made no attempt to apologize Could I make a suggestion There are a few points I'd like to make The police were called but no arrests were made I suppose we should make a start on cleaning this room Stop making such a fuss!
3
▶COOK◀
[transitive] to cook or prepare food or drink
When was the last time you made a cake John was making breakfast in the kitchen Who's going to make the tea
make somebody something
I'll make you some sandwiches
4
▶CAUSE◀
[transitive]to cause something to happen, or cause a particular state or condition
Its beautiful beaches make this a highly popular area with touristsIt was this movie which made him a starHis attitude made him very unpopular with colleaguesThe photo makes her look much older than she really is
make somebody/something do something
I like him because he makes me laugh
make something difficult/easy/possible etc The use of computers has made it possible for more people to work from home make something the best/worst/most expensive etc Over 80,000 people attended, making it the biggest sporting event in the areaThe President has made it clear that he is not going to change his mind
5
▶FORCE◀
[transitive] to force someone to do something
make somebody do something
My parents always make me do my homework before I go out
be made to do something
I was made to wait four hours before I was examined by a doctor
6
▶MARK/HOLE ETC◀
[transitive]to cause a mark, hole etc to appear
make a hole/dent/mark etc Make a hole in the paperThe cup has made a mark on the table
7
make it
a) to succeed in getting somewhere in time for something or when this is difficult
If we run, we should make it
make it to
With blood pouring from his leg, he made it to a nearby house
b) to be successful at something, for example in your job
He came to the US and not only made it but made it big (=was extremely successful) So far, relatively few women have made it to the top in the business world
make it as
He was told he had no talent and would never make it as a professional singer
make it to
England look less likely to make it to the finals
make it to manager/director etc How did anyone so stupid make it to manager
c) spoken to be able to go to an event, meeting etc that has been arranged
I'm really sorry, but I won't be able to make it on Sunday after all Nice to see you I'm glad you could make it
d) informal to continue to live after you have been seriously ill or badly injured
Frank was very ill, and the doctors didn't think he'd make it
e) to manage to deal with a difficult experience
make it through
I couldn't have made it through those times without the support of my boyfriend
f) used to say or ask what time it is according to your own or someone else's watch
What time do you make it I make it ten past three
8
make the meeting/the party/Tuesday etc
spoken to be able to go to something that has been arranged for a particular date or time
I'm sorry, I can't make Friday after all Will you be able to make the next meeting
9
▶ACHIEVE SOMETHING◀
[transitive] to succeed in achieving a particular position, rate etc
He was never good enough to make the team I don't think we'll make the deadline
10
▶GET MONEY◀
[transitive] to earn or get money
The plan could cost you more than you would make The business made a profit of £140 million last year His one aim in life was to make money She hopes to make a living (=earn the money she needs to live) from writing children's books He's made a fortune (=earned a lot of money) selling computers on the Internet
make something out of something
How easy is it to make money out of gardening
11
▶HAVE A QUALITY◀
[linking verb] to have the qualities that are necessary for a particular job, use, or purpose
+ noun
I'm sure you will make a very good teacher The hall would make an ideal venue for a wedding reception An old cardboard box makes a comfortable bed for a kitten
12
make it/that something
spoken used to correct what you have just said
Can we have two cups of coffee, please No, make that three
13
make do
to manage with the things that you have, even though this is not really enough
I hardly had any food in the house so I just had to make do
make do with/without
I usually make do with a cup of coffee for breakfast
For many people, make do and mend (=when someone manages with the things they have and does not buy anything new) was a harsh reality
14
make yourself heard/understood/known etc
to succeed in getting someone to hear you, understand you, or know that you are there
I had to shout to make myself heard above the music
15
▶BE A TOTAL◀
[linking verb] to be a particular amount when added together
Two and two make four There are nine people coming, plus me, which makes ten
16
▶CALCULATE◀
[transitive] used to say what you have calculated a number to be
I make that $150 altogether
17
▶SPORTS SCORE◀
[transitive] to achieve a particular score in a sports game
Surrey had made 92 by lunchtime
18
make somebody captain/leader etc
to give someone a new job or position in a group, organization etc
She's now been made a full partner He was made mayor in 1998
19
make believe
to pretend or imagine that something is true when it is not
I tried to make believe she was happy, but knew deep down it wasn't true
ￚsee also make-believe
20
make like
informal to behave as if something is true when it is not
He makes like he never met me before
21
make as if to do something
literary to seem as if you are going to do something but then not do it
She made as if to speak but then stopped
22
▶ARRIVE◀
[transitive] old-fashioned to arrive at or get to a particular place, especially when it is difficult
I don't think we're going to make the town before nightfall
23
make the papers/headlines/front page etc
to be interesting or important enough to be printed in a newspaper, reported on television etc
News of their divorce made the headlines
24
make or break
to cause something or someone either to be very successful or to fail completely
Critics can make or break a young performer
ￚsee also make-or-break
25
that makes two of us
spoken used to say that you agree with someone or that something that is true of them is true of you too
'I haven't a clue what's going on' 'That makes two of us'
26
▶MAKE SOMETHING PERFECT◀
[transitive] informal to make something complete or successful
The hat makes the outfit
27
make it with somebody
old-fashioned informal to have sex with someone
ￚsee also made
ￚsee also make somebody's day atday(19)
ￚsee also make friends atfriend(3)
ￚsee also make good atgood1 (35)
ￚsee also make sense atsense1 (5)
make away with somebody/something phrasal verb
1
informal to steal something and take it away with you
Thieves made away with the contents of the safe
2
old-fashioned to kill someone
make for something phrasal verb
1
to go in the direction of a particular place
ￚsynonym head forI think it's time we made for home
2
[not in progressive] to cause a particular result or situation
Both teams are on good form, which should make for a great game
ￚsee also made for each other atmade(6)
make somebody/something into something phrasal verb
1
to change something so that it has a different form or purpose
We can make your room into a study
2
to change someone's character, job, position in society etc
The movie made her into a star overnight
make something of somebody/something phrasal verb
1
to have a particular opinion about or understanding of something or someone
I didn't know what to make of her What do you make of the idea
2
to use the opportunities that you have in order to become successful
I want to make something of my life
make something of yourself
She has the ambition and talent to make something of herself
3
make the most of something
to get as much advantage as you can from a situation while you are able to
We've only got one day in Paris, so we'd better make the most of it
4
make too much of something
to treat something as if it is more important than it really is
It would be a mistake to make too much of these findings
ￚsee also make much of somebody/something atmuch2 (17)
5
make a day/night/evening of it
informal to spend a whole day, night etc doing something, because you have chosen to
We decided to take a picnic and make a day of it
ￚsee also make a go of something atgo2 (3)
ￚsee also make the best of something atbest3 (9)
ￚsee also see what somebody is made of atmade(4)
make off phrasal verb
to leave quickly, especially in order to escape
The men made off as the police arrived
make off along/across/through etc
The getaway car made off towards Horrocks Avenue
make off with something phrasal verb
to steal something and take it away with you
Thieves broke into the school and made off with computer equipment worth £40,000
make out phrasal verb
1
▶SEE/HEAR◀
make something ↔ out
to be just able to see or hear something
He could just make out a dark shape moving towards him
make out who/what etc
I couldn't make out what he was saying
2
▶UNDERSTAND SOMETHING◀
make something ↔ out
to understand something, especially the reason why something has happened
make out what/how/why etc
I couldn't make out what I had done to annoy her
As far as I can make out , he has never been married
3
▶UNDERSTAND SOMEBODY◀
make somebody ↔ out [usually in questions and negatives] to understand someone's character and the way they behave
Stuart's a strange guy - I can't make him out at all
4
▶WRITE CHEQUE ETC◀
make something ↔ out
to write something such as a bill or cheque
She was making out a list of people to invite The book gives advice on making out a will
make something ↔ out to
Make the cheque out to 'Spencer Cross Ltd'
5
▶SAY/PRETEND◀
make somebody/something ↔ out
to say that something is true when it is not
The situation was never as bad as the media made out
make out (that)
She always tried to make out that I was wrong and she was right
make somebody/something out to be something
He makes me out to be some sort of idiot
6
make out a case (for something)
to find good reasons that prove something or show why you need something
I'm sure we can make out a case for hiring another assistant
7
▶SUCCEED◀
especially American English to succeed or progress in a particular way
How did you make out this morning
8
▶SEX◀
informal especially American English to kiss and touch someone in a sexual way
9
make out like a bandit
American English informal to get or win a lot of money
The lawyers made out like bandits
make something out of somebody/something phrasal verb
to change a person or thing into something else
The Olympics can make sporting heroes out of previously little-known athletes
make something/somebody ↔ over phrasal verb
1
especially British English to officially and legally give money or property to someone else
make something/somebody ↔ over to
He made over the whole estate to his son
2
to change someone or something so that they look different or have a different use
Redgrave has made herself over completely for her movie role
ￚsee also makeover
make towards something phrasal verb
to start moving towards something
She made towards the door
make up phrasal verb
1
▶FORM/BE◀
make up something[not in progressive]to combine together to form something
ￚsynonym constituteWomen make up only a small proportion of the prison population
be made up of something
The committee is made up of representatives from every state
2
▶PRETEND SOMETHING IS TRUE◀
make something ↔ up
to pretend that something is true in order to deceive someone
I think they're making the whole thing up
ￚsee also made-up (1)
3
▶INVENT◀
make something ↔ up
to produce a new story, song, game etc by thinking
Nick made up a song about them When you're the boss you can make up your own rules I've given talks so many times that now I just make them up as I go along (=think of things to say as I am speaking)
4
▶PREPARE◀
make something ↔ up
to prepare something by mixing things or putting things together
I could make up a bed for you on the sofa Can you make up a bottle of milk for the baby
5
▶SOMEBODY'S FACE◀
make somebody ↔ up
to put make-up (=special coloured substances) on someone's face in order to make them look better or different
They made him up as an old man for the last act of the play One lucky winner will have the chance to be made up and photographed
ￚsee also made-up (2)
6
▶NUMBER/AMOUNT◀
make something ↔ up
especially British English to add to an amount in order to bring it up to the level that is needed
I saved as much as I could, and my parents made up the rest The company will be forced to pay $6 million to make up the difference
7
▶TIME/WORK◀
make something ↔ up
to work at times when you do not usually work, because you have not done as much work as you should
I'm trying to make up the time I lost while I was sick Is it OK if I make the work up next week
8
▶FRIENDS◀
also make it up
informal to become friendly with someone again after you have had an argument
make up with
Have you made up with Patty yet
Oh come on! Why don't you just kiss and make up
9
▶FROM CLOTH◀
make something ↔ up
to produce something from cloth by cutting and sewing
The dress had been made up to her exact requirements
make something ↔ up into
I plan on making that material up into a dress
ￚsee also make up your mind atmind1 (3)
make up for something phrasal verb
1
to make a bad situation better, or replace something that has been lost
The team will be anxious to make up for a disappointing start to the season I don't eat breakfast but I make up for it at lunch The good days more than make up for the bad ones
see usage note include
2
to have so much of one quality that it is not important that you do not have much of another one
make up for something in/with
What Jay lacked in experience, he made up for in enthusiasm She doesn't have a natural talent but she makes up for it with hard work
3
to do something to show that you are sorry for doing something that upset or annoyed someone
I'm sorry I was late To make up for it, let me treat you to a meal
4
make up for lost time
a) to work more quickly, or at times when you do not usually work, because something has prevented you from doing the work before
We rehearsed all day Saturday, to make up for lost time
b) to do a lot of something in an eager way because you have not had a chance to do it before
Palin didn't travel much as a young man but he's certainly made up for lost time now
make up to somebody phrasal verb
1
make (it) up to somebody
to do something to show that you are sorry about the problems you have caused someone
I'll make it up to you somehow He was looking for a way to make up to her for what he had done
2
British English informal to say nice things to someone or be very friendly to them in order to get an advantage for yourself - used in order to show disapproval
3
be made up to captain/manager etc
to be given a higher position in an organization
He was a security guard before he was made up to reception manager
make用法好多……不知道你是指最基本的，还是包括短语之类……这个解释大多数都挺好懂……不知有没有帮助~~
stop(verb)
1
▶NOT MOVE◀
a) [intransitive and transitive]to not walk, move, or travel any more, or to make someone or something do this
He stopped suddenly when he saw RuthStop, come back!He stopped the car and got outI was worried that the security guards would stop us at the gate
stop at/outside/in etc
She stopped outside the post officeA car stopped behind us
stop to do something
Sam stopped to give me a lift
stop and do something
He stopped and looked into her face
stop for
I need to stop for a rest
stop dead/short/in your tracks (=stop walking suddenly) Sally saw the ambulance and stopped short stop on a dime American English (=stop very quickly - used about cars) This truck can stop on a dime!
2
▶NOT CONTINUE◀
a) [intransitive and transitive] to not continue, or to make someone or something not continue
By midday the rain had stopped This is where the path stops The referee stopped the fight The doctor advised me to stop the medication People are fighting to stop the destruction of the rainforests
stop somebody doing something
I couldn't stop her crying
b) [intransitive and transitive]if you stop doing something, you do not continue to do it
stop doing something
I stopped digging and looked at him
What time do you stop workI've been smoking for over ten years, and I can't stop stop it/that (=stop doing something annoying) Come on, you two, stop it!Right, stop what you're doing and come over here
3
▶PAUSE◀
[intransitive]to pause in an activity, journey etc in order to do something before you continue
stop for
We stopped for a drink on the way home
stop to do something
I stopped to tie my shoe
stop to think/consider etc It's time we stopped to think about our next move
4
▶PREVENT◀
[transitive]to prevent someone from doing something or something from happening
The government tried to stop publication of the bookI'm leaving now, and you can't stop me
stop somebody/something (from) doing something
Lay the carpet on paper to stop it sticking to the floorThe rain didn't stop us from enjoying the trip
stop yourself (from) doing something
I couldn't stop myself laughingShe grabbed the rail to stop herself from falling
there's nothing to stop somebody (from) doing something There's nothing to stop you applying for the job yourself
5
▶STAY◀
[intransitive] British English informal to stay somewhere for a short time, especially at someone's house
I won't sit down - I'm not stopping
stop for
Will you stop for a cup of tea
6
will/would stop at nothing (to do something)
to be ready to do anything to achieve something that you want to achieve
We will stop at nothing to save our child
7
stop short of (doing) something
to decide that you are not willing to do something wrong or dangerous, though you will do something similar tha

材料和方法
11 材料
111 细胞及菌株 HAb18为分泌抗人肝癌mAb的鼠杂交瘤细胞株，由陈志南建株〔4〕；正常肝细胞株QZG引自中国科学院上海细胞库。pCANTAB 5E克隆及表达载体，为Pharmacia公司产品。
112 试剂 反转录系统为Promega公司产品；PCR引物： heavy chain primer 1和2(Pharmacia公司产品)，分别为重链5′端和3′端引物；light chain primer mix： 为10种轻链5′端和3′端引物的混合物，用来扩增鼠Ig VL基因；linker primer mix，为带有Linker序列(Gly4Ser)3的重链3′端和轻链5′端的引物，用来将VH，VL拼接成ScFv基因；RS primer mix，为带有SfiI位点的重链5′端引物和带有NotI位点的轻链3′端引物的混合物，用来扩增ScFv基因片段并引入酶切位点。引物A，D分别为重链5′端引物(含EcoRI酶切位点)及轻链3′端引物(含SalI酶切位点)〔5〕。A，D引物的序列如下:
引物A VH-1（正向）:
5′ GTGAATTCATGCAGGTGCAGCTGTTGGAGTCTGG 3′
EcoRⅠ
引物D VL-2（反向）:
5′ CAGTCGACTTACGTTTGATCTCCAGCTTGGTCCC 3′
SalⅠ
12 方法
121 总RNA提取、cDNA第1链合成及VH和VL基因的扩增 取对数生长期的HAb18细胞，用异硫氰酸胍变性法，提取细胞总RNA。参照Promega公司反转录系统的产品说明书，反转录合成cDNA第1链。将cDNA第1链合成反应物分为两份，分别加入VH和VL引物各2 μl进行PCR。
122 ScFv基因的组装 回收的VH和VL基因片段与linker primer mix等摩尔混合，加入dNTP至终浓度为25 mmol/L，Taq DNA聚合酶5u，进行7次PCR循环(94℃ 1 min，63℃ 4 min)。将VH和VL基因拼接为ScFv基因。在上述反应体系中，加入RS primer mix，再进行30次PCR循环，可在ScFv基因的两端分别加入SfiI和NotI酶切位点。
123 ScFv基因的克隆和筛选 上述加端PCR产物经18% 琼脂糖凝胶电泳，玻璃乳回收后，用SfiI和NotI消化，并与以此两种酶双酶切的pCANTAB 5E噬菌粒DNA连接，转化Ecoli HB2151，在SOBAG（含100 mg/L氨苄青霉素和2%葡萄糖的SOB培养基）琼脂板上筛选转化菌。从转化板上挑取单个菌落，用2×YTAG（含100 mg/L氨苄青霉素和2% 葡萄糖的2×YT培养液）于30℃培养过夜，以碱裂解法小量提取噬菌粒DNA，以引物A，D进行PCR扩增出ScFv基因片段，并克隆入pUC19载体。
124 ScFv核苷酸序列的测定 以美国PE317-A型自动序列分析仪进行序列测定。
125 可溶性ScFv基因的分泌型表达 将含ScFv基因的重组噬菌粒菌种接种于5 ml LB 培养基，过夜培养。加至50 ml SBAG(含100 mg/L氨苄青霉素和2%葡萄糖的SB培养液)中，30℃振荡培养1 h，5 000 r/min离心15 min，弃上清。将沉淀重悬于50 ml SBAI(含100 mg/L氨苄青霉素和1 mmol/L IPTG的SB培养液)中，37℃诱导3 h，离心同上。取沉淀悬于5 ml新配制的溶菌酶（1 g/L），20% (w/v)蔗糖，30 mmol/L Tris-Cl(pH80)及 1 mmol/L EDTA (pH80) 溶液中，冰浴10 min，4℃以12000r/min离心5 min，上清即为外周质部分。将其冷冻干燥后，贮于-20℃。临用时以001 mol/L PBS溶解至500 μl。
126 ScFv基因可溶性表达产物的鉴定 采用SDS-PAGE及Western blot(二抗为鼠抗-E-tag抗体)进行鉴定。
128 流式细胞仪分析ScFv的活性 收集培养的肝癌细胞SMMC 7721，正常肝细胞QZG及胃癌细胞SCG 7901，用10%FCS RPMI1640调整细胞浓度至5×106/L~1×107/L。每管加入40 μl细胞悬液，100 μl ScFv表达产物，再加入50 μl 1∶20灭活正常兔血清（用DPBS稀释），于4℃作用30 min，洗涤2次，以800 r/min离心5 min，弃上清。加入抗E-tag抗体8 μl(25 g/L)，4℃作用30 min，洗涤2次，弃上清。加入50 μl FITC标记的羊抗鼠抗体，充分振摇，4℃作用30 min，洗涤2次，加入500 μl固定液，以流式细胞仪进行分析。同时设正常肝细胞、胃癌细胞为阴性对照组，肝癌细胞加亲本抗体Hab18为阳性对照组。
2 结 果
21 VH和VL基因的PCR扩增及ScFv基因的拼接 VH和VL基因测序结果显示，VH为363 bp，VL为342 bp，与预期的VH和VL基因片段大小相符。将VH和VL基因用linker连接成ScFv基因（其中linker为45 bp）。以此为模板加入RS primer mix进行PCR扩增，所获扩增产物的大小约为730 bp，表明VH，VL及linker primer mix的浓度配合准确，并引入了SfiI和NotI酶切位点（图1，见第Ⅳ页）。
22 ScFv基因的克隆 将ScFv基因克隆入pCANTAB 5E中，连接物转化EColi HB2151。在SOBAG固体培养基上筛选转化的菌落。然后随机挑选8个菌落，小量制备噬菌粒DNA，以Sfi I 和NotI双酶切鉴定。若含有外源插段者，应切出约730 bp大小的片段。结果表明，8个克隆均含插段。
23 ScFv基因的序列测定 如图2所示，ScFv基因全长为726 bp，包括预期的VH，VL基因和linker序列。VH基因序列处于linker的上游，VL基因序列处于linker的下游。
24 可溶性ScFv蛋白的鉴定 可溶性ScFv-E-tag融合蛋白，预期的Mr为29 000。E-tag可作为蛋白标签，通过鼠抗E-tag抗体来检测ScFv基因的表达。将阳性克隆以1 mmol/L IPTG诱导3 h，对IPTG诱导和未诱导的菌体进行裂解，经SDS-PAGE后，在(Mr为29 000处多出1条明显的新生蛋白带，与预计的ScFv-E-tag融合蛋白的大小Mr为29 000)相符（图3，见第Ⅳ页）。同时做与图3相同的SDS-PAGE并进行Western blot，在Mr为29 000处有显色条带，即为能与抗E-tag抗体特异性结合的可溶性ScFv蛋白（图4，见第Ⅳ页）。
25 可溶性ScFv蛋白与肝癌细胞的结合活性 以可溶性 ScFv蛋白为一抗，抗E-tag抗体为二抗，FITC-抗鼠IgG为三抗，用流式细胞仪测定荧光标记细胞的阳性率(图5，见第Ⅳ页)。肝癌细胞组： 无关抗体(抗-CD3)为16%，亲本抗体为975%，ScFv 为309%；正常肝细胞+ScFv蛋白为26%，胃癌细胞+ScFv蛋白 为41%。表明可溶性ScFv蛋白能与肝癌细胞特异地结合，而不与正常肝细胞和胃癌细胞结合。荧光显微镜观察，可溶性ScFv蛋白与肝癌细胞的膜抗原相结合，在细胞膜上可见颗粒状的荧光(结果未显示)。
3 讨 论
随着Ig基因结构与功能研究的不断深入，基因工程新技术的不断涌现和成熟，基因工程抗体的研究取得了很大进展。现获得的各种基因工程抗体已基本克服了鼠源性mAb的缺点，及制备人源性抗体的困难，使之在基础医学研究和临床疾病的诊断、治疗和预防等领域，特别是肿瘤的治疗中，得到了极为广泛地应用，有的已进入Ⅲ期临床应用。运用基因工程重组技术，将鼠源性抗体恒定区和可变区的框架区以人源性相应抗体片段取代，可降低抗体的免疫原性，有效地减轻免疫排斥反应〔6〕。利用基因突变技术改变抗体的某些结构，则可提高抗体的亲和力，延长其半衰期〔7〕。另外，还可将抗体基因与其它功能性基因相连接，赋予抗体新的功能。
近年发展起来的抗体库克隆技术，可无需进行细胞融合制备杂交瘤，甚至无需进行免疫，即可直接从基因水平上，获得所需的针对相应抗原的抗体。制备基因工程抗体的关键，是获得抗体可变区基因，拼接成单链抗体基因，并表达出有生物学活性的产物。
本文从分泌抗肝癌mAb的杂交瘤细胞(HAb18)中，用RT-PCR，克隆出抗体可变区基因，其关键是PCR引物的设计。大多数研究者设计合成的引物，通常是针对V区FR1 5′端（或信号肽）的保守序列和J基因3′端序列（或恒定区基因序列）而设计的，特异性不够强，不能有效地扩增某些目的基因。Pharmacia公司噬菌体呈现系统中的混合引物，系针对鼠Ig基因不同家族的序列而设计的。从理论上讲，适用于所有类型的Ig，每种可变区基因都可获得扩增，对全套抗体库的构建非常重要。
本研究所用pCANTAB 5E是Pharmacia公司的产品，含有氨苄青霉素抗性基因，plac启动子和M13噬菌体基因间隔区片段，在多克隆位点与gⅢ编码区之间，有1个c-myc tag序列和琥珀终止码TAG，IPTG可诱导外源基因的表达。在不含有琥珀抑制基因的菌株（如HB2151）中，E-tag后的终止码被识别，多肽链的翻译终止于此，从而可生成具
有生物学活性带有13肽E-tag的可溶性ScFv蛋白，释放于细菌周质中。E-tag为源于人c-myc基因的一段序列，对ScFv基因的结构和活性均无影响，可作为可溶性ScFv蛋白的标签，用抗E-tag抗体来检测ScFv基因的表达并纯化其产物。我们用含有ScFv基因的重组噬菌粒载体转化大肠杆菌HB2151，经IPTG诱导获得可溶性表达产物。用抗E-tag抗体做Western blot证实，表达产物为ScFv-E-tag融合蛋白。流式细胞仪分析表明，此ScFv融合蛋白可与肝癌细胞结合，荧光标记细胞的阳性率为309%，而不与正常肝细胞及胃癌细胞相结合，表明此ScFv蛋白具有肝癌结合的特异性。
参考文献
1 顾方舟，陈妙兰，陆如山等 肝癌研究概况与展望 北京： 中国协和医科大学，北京医科大学联合出版社，1993: 1-3
2 Hoston JS，Levinson D，Mudgett HM et al Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Ecoli Proc Natl Acad Sci USA，1988；85: 5879-5883
3 Kroesen BJ，Wellenberg GJ，Bakker A et al The role of apoptosis in bispecific antibody-mediated T cell cytotoxicity Br J Cancer，1996 ；73(6): 721-727
4 陈志南，刘彦仿，杨继震等 抗人肝细胞癌单克隆抗体的产生及其相应抗原的免疫组化定位研究 单克隆抗体通讯，1989；5(2)： 33-36
5 杨安钢，吉昌华，韩 骅等 抗人CD8单抗κ轻链可变区基因的克隆和序列测定 生物化学与生物物理进展，1992；19（4）： 286-288
6 Man SC，Jeanne BK，Bednarik EJ Humanized anti-Lewis Y antibodies: in vitro properties and pharmacokinetics in rhesus monkeys Cancer Research，1996；56: 1118-1125
7 Mats O，Henrik O，Michael M et al Light chain shuffling of a high affinity antibody results in a drift in epitope recognition Mol Immunol，1996；33(1): 47-56
(收稿 1998-03-03 修回 1998-03-26)

首先你在後台判断用户输入，如果title为不空、notiice为空，那就用sql = "select from notice where title = 'title' '，

如果是title为空、notice不为空，那就用 sql  = "select from notice where notice = 'notice' "；

如果两个都为不空（就是用户两个关键字都输入了），那就用sql = "select from notice where title like '%title%'  and notice like '%notice' ";

如果都为空，那就查找出所有或者什麼也不查找。

=========================================================

模糊查找的方案有很多种，你可以，比如“全匹配”就是完全等於、"区分大小写"等等。看你自己设计。用like是模糊查询，用“=”就是全匹配，你可以为用户增加这些选项。多个关键字一起查询，就用"and"或者"or"来连接就好了。

还是不懂的话，建议重新学一下sql语法语句吧！

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