                      ˿µϣ20050710գ

05.07.09ûи
05.07.10, hbchendlԲ԰桷
05.07.10, ˮԲ԰ļ㿴
05.07.10, Ϊ˽һЩǵݺҲı䣿ǻȺʿ
05.07.10, ϾҵѧһʦϴŽڡܡ
05.07.10, guayfzɲʿԾ֮絽סɱ˭
05.07.10, իǳо
05.07.10, TonyҲ̸̸ҵġϷá
05.07.10, ¸աигڵ⡷
05.07.10, ˫ڡйĿоѶȥˣ
05.07.10, ͡йҽϵͳĸܡ
05.07.10, Ƿݵߵֱˡĳо
05.07.10, ϷĩƷ籩У

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Բ԰

hbchendl

    廪ѧĻóѽóôĹ
棬Щط˺Ϳ

1.      ǰеķ̵׶Ի˶ƻ?

    ԭгô(ڶµĽ)

 210 Ӱʶ

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̬

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رˮ

ˮ         -S1                  +L2              -L2
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ˮ

ˮ          0                    0                  0
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ַ     -S1                  +L1             +L1

עС+ʾӰ죬-ʾӰ죬SʾӰ죬L
ʾӰ죬 2ʾӰ1ʾӰһ㣬0ʾӰ졣

￴ʲô? һʩڶԻиӰ졣ҪǶˮ
̬׵Ӱ졣Ӱԭʵܼ򵥣˺ˮ
ڳ˺׵࣬ӰŹ֡Ҫע⣬ʩڵӰ죬Ƕڵģɻ
ġĿֻһԵӰ졣ʵǵ˵ʩڼԻӰ
ƻ 

ʵҪʩڼЧӱпԿ
ֻҪˮ֣˵ˮеˮһ⣬ĿӰ졣
ˮֵĻŻԻɲӰ졣ֻصˮ⣬
Ϳٲˮ͵öಹˮˮô˵Ҳ
ƻĴñѽ

ҾͿ˵̡ƻӰ컷ĸ
?

2.      Բ԰ĺ׸òýз?

ƺѾǾӦýз˸βͣ˵
ͬҪԴһҪһһ
ͬʱҪˮశһЩߵֲ

ҵȻûʸȥ˵ﲻǱΪĿĲ
Ϊ˽ʡʽ𣬶Ϊúˮһ㡣һˮǶ? ȫ
Ԥƣȫ겹ˮ350򷽡ķԤƣ400򷽡
50򷽡50ˮԲ˵һʿ֧ˮ˵
ǶӵĸԺͱеĵˮλӰ? 
˵ʵʩǰˮλĳ½Բ԰ˮ
©Ϊ363.4 m3/aˮ©Ǳˮλ̧0.1mϣӰ췶
ΧԼΪ14km2©ֱռкͺĵˮԴ0.14
1.5%ˮˮλӰ췶Χֱռкͺ0.06%2.35%
ʵʩ󣬾һˮҲ©ˣԱкͺĵˮˮλӰ
̶ȲҲǧ֮һͰٷ֮ҡ

ֿˣΪٷ֮ĵˮλӰ죬ֵøôĴȥ
𲿷ַĤҪԺÿ50ˮ?

УˮĲԺֲôҪ? ݵķеǣ
ˮַǲ˷ˮķ֮һ?

3.      ΪʲôҪĤճ?

дƪ֤Ĥܣҿܶģһ
ǷĤ޶޺ܴﵽʳƷװİȫ׼ǷĤ30굽50
ϡΪʲôҪأҴӱֻҵһ: Ŀǰûʹø
ĤϲԵıۺϻѺԵȻϡ
ǵԲ԰ǹصﱣλַ԰ԣӦȲ
ķϺʹʩ

ۺϻѺǵһμʲôۺϻѺһǲ

ճҲǷҷЧ̷ĤͬʱճʩҲһҪ
ɺˮڳ࣬ƻһ»һ: ճӺζ? Ƿֵô
𴦵ȡ? ֪שҵԭճÿ
ΪԲ԰ĻҲƻһ±ﻷǲеڶǽ
ǽ˼?

֮⣬ҪһѾ̺õĤڳ˷ѵǰڵĸһ
ΪһۺϻѺԡ ˵ɡǲ̫? 
ָܷĤĿǰδ֤жԣȻȻ
Ĥ̬Ҫ󡱡 ôȻȻ? Ȼ
ǲ롰ĪС֮ͬ? ο˵ǡӦȡá
ǡ롱áδĤֱճҲ˵ùȥĤѾ̺ˣ
ʲôҪǵò˷һʽ𲻿?

֮档ǰһֶԷ̵Ļ̳
ʩǰδ֮⣬ûʲô⣬ҲԻʲô
棬ȴðƻҪġĵֶκĿ
ĲҪѷĤܵĻճȥƻĵط̬
Ҫÿ50ˮⲻڻ? 

(XYS20050710)

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Բ԰ļ㿴

/ˮ

зԲ԰˴𰸡Ȼû뵽ǣ
ĤҥԣĿѧ۷֮󣬹һȴһ
뻷ȫ෴ķ

ݱһֳָܾŽܾͨ75֯
רҶ廪ѧĻ飬ͬñۣҪԲ
԰׷̱ȫġ

˵ָܷĤĿǰδ֤жԣȻ
ȻĤ̬ҪںߴĤȻܹγ
άֽϴˮ򾰹ۣڶʹˮԻָ谭ˮ彻
ͲԺ׺ͺߵֲӰ졣ͬʱڹ
̽ȱЧıʩˮ̬ϵͳƻˣ
̴ĤǲѧģҲǲӦᳫģȫġ

ΪһֵĴʩһЩ֮ʵʩĴʩ
Υ˻ĽҽҽԲ԰ɾ޴ʧ
㣺

1˾ֳ˵谭ˮ彻ͲԺ׺ͺߵ
ֲӰ졣

ٵˮΧֲǱȻģǣԲ԰Բ
ûʲô⡣Բ԰ƽһ׵ĸ㣬ֲ
ҪԲ԰ûзĤõʯͷɵĺ©٣
Χֲﹹвһڱκһˮ嶼Եõ֤˵
©Ƿ븶ĴۣһͲܸˣ˾
˵ǿЧӦȫķ˻϶ҪԵĽ
ۡ

2ֳ˵ڹ̽ȱЧıʩˮ
̬ϵͳƻ

ע⻷СԭƺͽС仰ǳ
ָʩɵνƻǷ̬ƻ򣬻
ڷԼĽۣܶαӶǲûǷǵİ취
Ľ󵼹ڡȻ˾ֳԻҲ⡣
뻷Ľȫ෴ġ⣬νʩɵġƻ
ҲԵģ廪ѧҲ᲻ûκ̬ƻʩ취
ڣȻ󲿷ʩѾɣΪ˱ʩƻӦþٵĸı
ԭеʩֻµġƻˣ˾ֳġù̴
ĤǲѧģҲǲӦᳫģȫġǶԻ
⣬һʵʩ⻷Ľ飬Ҫʩ
̬ɵĲɱƻ

3ֳǿĴʩҪϵصĵˮҪͻ
Ӱȶأʵǵؿǹ̵ĶӰͳЧͳ价
úɱ
һԲ԰δʵʩ׷̵򣬲Ĥȫ
ȡȻճ
ڶ粴԰ˮ⣬̵ķĤӦȫճԭ׵
ࡣ߲Ĥ
԰׸40.7׵ҪĤճ
ҲĤ
ģԸѾķĤȫ졣ɰʯΪĻҪ
ȥɰȻճԭ׵Ҫȫͷܱ
10⣬ĲӦĤԱֵ֤ĲͬʱΪ
άԲ԰ˮ̬ϵͳҪͳ滮԰ˮˮ
ˮ֤ˮˮʣ԰ڵˮҲҪϼӱֹȾ

Ϊ˾ֳļ鶼Ǹݣһ䡰Ŀǰûʹø
ĤϲԵıۺϻѺԵȻϡ
ġʵǶԻһ⣬ѺԵ֤͡
ʩһҪʹȻϣԲ԰̣˹Ĥ
Եơ

AĤķЧҪճõö࣬ͨ켼Ϊ
ЧκȻ϶޷ʵֵģҲΪʲôȫ綼ձ
ĤҪԭ

B˹Ĥɱ͡κηĲöܲǳɱء
Բ԰˵ճĿԳЧĤ֮⣬ճĳɱҲ
һҪءԲ԰ûճôԶȡʲôճ
ֱǡԽˮĴʩ

CճЧϲһҪȽϺճ𵽽Ϻõķ
ãԣճĹʩýңΪʵ
ЧҪѹճ㡣ˣճһʺϱ
Ĥȱǲڱ븲һ㡣ǣڷĤ
ϿԸһ׺㣬ǡǡǳԲ԰ˮҪֳˮҪ
ԣϿԲ԰÷ĤǺ̬ЧҪճЧǿ
ࡣ治֪֤Ķʲôרҡרҵô
һⶼ㲻

DûвģȻֵļٵˮزĸ
ã÷Ч̫õճΪָճǷǳ
޴ѡԲ԰Ҫֲ÷Ч̫ǿ뱣һ
ˮ͸Ĥ¸ʲôճֻҪʣµĺ
ײֲĤɴﵽЧòʣֻԭеķĤ
ϴ򼸸վͿԴﵽ©ĿġΪȻ϶Ҫ
ӦòȡӦĴʩҪȫ𱸵ʵǲܰ쵽ġ

4һֲԲ԰Ĺţֻ档
֯רҶԻĿѧԽ֤飬Ƿ׼棬
ԻĽûȨԲ԰ʲôĴʩΪ
ĴʩĿԸ磻˭֪ӱͭҪãʹ
ʲôǿͻҪԼĲе顣ĴʩͺñȻ
һԲ԰ýƵĻ⣬ֲͭǮⲻ
ǷΪ˼ң˳𣿻ִ˾پˡʵϻ־Ӧ
黷ϲϿѧȷԲ԰Ի߲ԸĤǷ
̬ƻڸ㲻ʲôϷӦԲ԰
ܲŵ飬ûȨʲôĴʩΪĴʩӦ
߱йزţټרڻĻȷžʵ
ʿɲԡ

5⣬ʵһֱΪ˵Բ԰Υûзݵġؼ
άĿڽĿòŻ񣬳Բ԰ȫ
άĿʹҲڴޣû̳ҲϾҪ
ޣҲûȥĿûңںͽȻ
ҪΪʲôһֻҪԲ԰⣿ǲǻΪ˰α
ҥΪͣѰҽڣ˻÷ΧҸ
Ϊǵ͵Υһƪڻɵ£ڼȻѾ֪
ַ֧֮ȻܽĻҿΪ˽⣬
ҵʧԲ԰ӦôӺĿĳ߹һ֣ͨ
⡣֮ҪԺִУȻָԲ԰ʩ
ˮԲ԰Ϊ̬ͣƻ״

(XYS20050710)

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Ϊ˽һЩǵݺҲı䣿ǻȺʿ

nanyaoda

꽭ʡйҩ﷽б֪꣬˻ȺʿĴļ
νݺмƣǲ֪Ǹʲôģˣǲѯй
ıȻ

ѧӦóš

Ͼѧҳ־Ϸרҵ40ƪ
Ҳ֪δܲ鵽ĴϾѧҳṩ1987
ѧʿѧλҮ³ѧоں˾
BayerҩоΣ߼оԱͨISI Web 
of KnowledgeѯõĽΪPaiva AA, Tilton RF, Crooks GP, 
Huang LQ, Anderson KS. Detection and identification of transient 
enzyme intermediates using rapid mixing, pulsed-flow electrospray mass 
spectrometry. BIOCHEMISTRY 36 (49):15472-15476,1997.Author Addresses: 
BAYER CORP, DIV PHARMACEUT, W HAVEN, CT 06516 USA, YALE UNIV, SCH MED, 
DEPT PHARMACOL, NEW HAVEN, CT 06520 USAһƪġںδҲ
 ISI Web of Knowledgeվĸ̫С

ҲԸı䡣

Ͼѧ 2005420աϴ󺣹߼רһ
Ⱥڡд1977꣬22ĻȺӹͨϾΪָ߿
ϴһѧеһԱôƲȺʿӦó1955ꡣǣ
200412¹ġʡ˲Ÿ߷塱ڶĿУ
Ϊ45ꡣǺιʣҲɱ仯ʡ˲Ÿ߷塱Ƿ涨
䳬45˲룿ѧĿߵĲΪδΪ3
ԪĿֵα

ݲϱ仯

Ͼѧҳܣҩҩṹоֿ
ʮҼҩƳһ࿹ҩǲ֪ںδˡʮ
ҼҩϾѧ 2005420աϴ󺣹
רһȺڡд20035£ȺƸΪϴְڲϴ
ҩﻯѧ˺ҩﻯѧԱʮֹҩ
ʵڲ֪һҩҪʱ䣿Ҫ룬
༸оǽյҩ￪⡣˽⵽
оָϾѧѧѧԺҩﻯѧоϾѧѧѧԺҩ
ﻯѧоӦϴҩﻯѧоͬɣ

Ͼѧҳܣ1987ѧʿѧλҮ³ѧ
оں˾ݶBayerҩоΣ߼
оԱ֧GLPʵҩ,ҩЧ,ҩоҩṹⶨȡ
2003ƸΪϾѧʱںͲʿʦϾѧҩﻯѧо
2004ȫʱعϾѧҽѧԺԺҩﻯѧо
ڣҮ³ѧоڣݶBayer˾߼оԱ
ͬʱƸΪй廪ѧѧԺʿʦйҽѧѧԺڡ
ϾѧƸдšϾѧҽѧԺԺΪαˡԺ
ѵϾѧƸдˣ

2004 ͨݾÿվ2004-05-26Ͷ1200עʱ500
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ҩҵ޹˾Ӣعɹ˾˰족ù˾滮õأĶ
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Ӣعɹ˾³Ⱥʿ˵"֮ѡͨͶʣΪ
Ǹܵͨίֳļ⡣"Ͼ
ѧҳΪβΡӢعɹ˾³һ£
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Ͼѧ 2005420աϴ󺣹߼רһ
ȺڡдһͶԵǻ⣬йĻĻȺ
ݶ˾߲ͬǾȵľعȫְϴڣϴڵ
Ү³оһְ200531գϲѯȺʿѱƸΪϾ
ѧҽѧԺԺǣϴƼ 200577ձڡѧʿվ
׳ᡱϣĴҽҩѧ˾ݶ˾Bayer
ҩоҩṹѧоΡ߼оԱ1982ҵϾѧ
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ѧӽĽйŴʺѧ
ѧ̳2005425-26ڹֳɹСڻ
ĵĻȺʿҩоеĵѧоΪЩ
һֲϾѧϾѧȫְڣΪλ衰Ӧϴѧʿ
վ룬پҵѡĹѧӽĽô
˵ϾΪֱ˰ݶ˾Bayerҩоҩ
ṹѧоΣݶBayerҩоݶ˾
BayerҩоҩṹѧоҲӦòͬɣ죬
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ڡϴ󺣹߼רһȺڡһжα
ԼĸУᡣǣΪζദƣ氮ǹ


Ͼѧ 2005420աϴ󺣹߼רһ
Ⱥڡд1987꣬ȡòʿѧλоҮ³
ѧҪоɫıֵõ˵Ͽɣڸ󲿳Ƹ
ǩȺ䷢ˡJ-1ܳ˲̿ڸ󲿳ǩ
J-1ܳ˲̿ѵҪ˴ǩ
ȵȡźڣ

ϾѧͽʡƼʵʹ

һϾѧҳܣ

ҩҩṹоֿʮҼҩƳ
һ࿹ҩĿǰҩѽٴо״ִ׷
СӼĵҩ·ִۣΪļ
ѧоƽ̨־Ϸרҵ40ƪ2000ðݶ˾
ƼһȽ199611ܹȰ룬Ϊѧߣܵ˹
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ձҪý屨ĿǰѳеҼʡĿ6ش
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1982ɼҵϾѧѧϵͬ기ѧ1987
ѧʿѧλҮ³ѧоں˾ݶBayer
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ҩоҩṹⶨȡ
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2004ȫʱعϾѧҽѧԺԺҩﻯѧо
ڣҮ³ѧоڣݶBayer˾߼
ԱͬʱƸΪй廪ѧѧԺʿʦйҽѧѧԺ
ڡ


ִ챨2005꣩߼רһȺϴְط
ÿ˾
챨ͨѶԱ޾
գһϴѧӵ˾߹ܵĴҵľݽϾ
ѧٿݽǰݶ˾Bayerҩоĸ߼оԱϾ
ѧҽѧԺԺҩоȺʿ
ȺϾѧ77Уѡ1982기ѧعǰĴҽҩ
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רҵרҵҪǿרҵسԽԽǿ
Ҫ׷Ҫġ;ֵףԶչѧϰǴ
ҵ۴ںֽ׶ΣֻнڵĿ꣬ҪóԶһЩҪ
Сڵļƻ룻ֹۣԴۣʱҪ
һ棻ǽʼɣڴ˾£ܶ鲻ܼ򵥵رΪ
븺˼ʹϵҪķʽҪɵֶμЭ
ԣƸԱԱǷʱȽϿǷдǷ
нȡĺԡ
ɹıر
׺רҵĻȺидһʮ
˲ܺܺõദȱŶӾ񣬼ʹҲᱻ˾á
߱׺֮רҵڶľӱ
ԷҲ绢Ⱥ19825ʱ
òʿѧλ1987꣬ȡòʿѧλоҮ³
ѧҪоɫıֵõ˵Ͽɣڸ󲿳Ƹ
ǩȺ䷢ˡJ-1ܳ˲̿ 



ϴ󺣹߼רһȺ
ԴϾѧ 2005420յ7  ܵ885 
	ϾѧƼ¥19һ칫Ⱥоر̹
ԶϤĶаεضĸ¥㳤һƬⷽϤУ԰
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Ժҩо
	һع飬Ǽ򵥵űЧĺԸǴݰѰҵ
ζƮ֮滺½̹
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	1977꣬22ĻȺӹͨϾΪָ߿ϴ
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йٰ"и"ԣѧѧΪĸ￪źһо
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оȺɫıֵõ˵ϿɺӣΪסƲʿ
ڸ󲿳Ƹǩ𣬰䷢й½ĻȺJ-1ܳ˲̿
	İؿٳͷƤй˶ɧأǻ
ȺԸ
	ϣûгԹ"˾ƥ"һ࣬Է˾ƥ
Ϳҵݶ˾ÿһ˶йٵĹ1990꣬
Ү³ѧİݶоƸ߼оԱֻ1
ˡϸѡȺڶྺӱ״
һλ֡

	Ϊݶ˾ĸ߲ܣǻȺɹҲһ
˵ĳɹһ壬뵱󲻾ãӵ20
ԪнĻȺڿҿһ߼סլһ׸߼Ͼʱ⣬
Сɽϵ⴦̨ңԶ͵֣ľס
ĶڲƺϾ(תİ棩Ͻӵһ棩ˣ㱼䣬
婺Ӱߣŵ婺ˮϢ婺ĽͶʱĸ׵ĺ
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	ڻȺйĸ￪ߵֱߣҲǹͬ
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	ݶоһйģȺҪԱθ
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Title: Detection and identification of transient enzyme 
intermediates using rapid mixing, pulsed-flow electrospray mass 
spectrometry 
Author(s): Paiva AA, Tilton RF, Crooks GP, Huang LQ, Anderson KS 
Source: BIOCHEMISTRY 36 (49): 15472-15476 DEC 9 1997 
Document Type: Article 
Language: English 
Cited References: 14      Times Cited: 21          
Abstract: Rapid chemical quench methods coupled with off-line 
detection have proven to be very useful in identifying enzyme reaction 
intermediates, However: a limitation to this approach involves enzyme 
intermediates which are too labile under the chemical quenching 
conditions to allow detection and characterization, In this report, we 
describe the development of a novel approach for the detection and 
characterization of enzyme intermediates on the subsecond time scale 
using a ''pulsed flow'' method which employs a direct Interface 
between a rapid-mixing device and electrospray ionization mass 
spectrometry. The application of this technique with the enzyme 
5-enolpyruvoyl-shikimate-3-phosphate (EPSP) synthase is demonstrated, 
This enzyme converts shikimate-3-phosphate (S3P) and phosphoenol 
pyruvate (PEP) to EPSP and inorganic phosphate. Previous rapid 
chemical quench studies have shown that this reaction proceeds through 
a tetrahedral intermediate [Anderson, K. S., rt al. (1988) J. Am. Chem. 
Soc. 110, 6577-6579] formed transiently at the enzyme active site, We 
have shown that this tetrahedral intermediate can be directly detected 
on a subsecond time scale 1 without chemical quenching by interfacing 
a rapid mixing apparatus directly with an on-line electrospray 
ionization ion trap mass spectrometer. Negative ion mass spectra 
collected by electrospray ionization indicate peaks for S3P (m/z 253), 
PEP(m/z 167), EPSP (m/z 323), and the tetrahedral intermediate (m/z 
421). Further confirmation was provided by performing the same 
experiment-with [C-13-1]-labeled PEP, These spectra confirmed the 
anticipated shift of 1 atomic mass unit for PEP (m/z 168), EPSP (m/z 
324), and the tetrahedral intermediate (m/z 422) with no change in S3P 
(m/z 253). Thr collision-induced dissociation of the unlabeled 
tetrahedral intermediate peak (m/z 421) produced a daughter ion at m/z 
323, which is most likely EPSP resulting from the loss of phosphate 
and is consistent with previous studies which have examined the 
chemical breakdown of the tetrahedral intermediate In solution 
[Anderson, K. S., et al. (1990) J. Biol. Chem. 265, 5567-6672], This 
technique is under development and should be a useful method to study 
the transient formation of enzyme intermediates. 
KeyWords Plus: TETRAHEDRAL INTERMEDIATE; EPSP SYNTHASE; IRON 
BLEOMYCIN 
Addresses: BAYER CORP, DIV PHARMACEUT, W HAVEN, CT 06516 USA
YALE UNIV, SCH MED, DEPT PHARMACOL, NEW HAVEN, CT 06520 USA 
Publisher: AMER CHEMICAL SOC, 1155 16TH ST, NW, WASHINGTON, DC 
20036 
Subject Category: BIOCHEMISTRY & MOLECULAR BIOLOGY 
IDS Number: YL105 
ISSN: 0006-2960 

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ܶòö⣬һ޳ܲ޳ܵ⡣

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ڶԽڵ, Ӧð/ѧҵĳ·. ˵ѧ
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Ҳ̸̸ҵġϷá

Tony

źĺ,ԼⱲûᱻܹȺ,û
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Ƽ䣺yahoo.com, hotmail.com, gmail.com

ġ˿ʼбĵ䷢һհż

xys-subscribe@yahoogroups.com 

塢ͣ˿ʼбͣ䷢һհż

xys-unsubscribe@yahoogroups.com 

Ͷxinyusi@yahoo.comôıļͶ塣

˿(www.xys.org)(xys.dxiong.com)(xys.3322.org)(xys.dyndns.info) 
