Cch vit cng nghin cu khoa hc (GS Nguyn Vn Tun) i vi nhng ngi nh ti, vit cng nghin cu gn nh l mt ngh.

gh. Nm no cng phi vit t nht 3 cng, c khi l n xin bt, cng c khi l n xin ti b nhim. Vit rt nhiu v tht bi cng rt nhiu. Tht bi nhiu n ni kh m ht! Tht bi gn nh l mt qui lut! Nhng cng c thnh cng, d s ln thnh cng t hn s ln tht bi. Chnh qua nhng tht bi, ti mi hc c nhng bi hc au lng, v l l do ti sao ti mun chia s cng cc bn tr hn, hay cc bn cha c kinh nghim (hay c t kinh nghim) v cch vit cng nghin cu. C nhin, ti khng dm ha nu cc bn tun theo nhng g ti hng dn l s thnh cng, nhng ti dm ha l xc sut thnh cng s cao hn l khng lm theo nhng hng dn y. Trong cuc sng hng ngy, k c cng vic chuyn mn, bt c ai trong chng ta thng gp nhng vn ng tm hiu, c khi rt l tng cho nghin cu khoa hc. C nhng vn khng hn l phc tp, nhng c khi li rt n gin. Thng, nhng vn n gin l nhng vn kh nht, v c th dn n nhng khm ph quan trng. Ti sao nam gii hay cht sm hn n gii? Ti sao ngi dn vng nng thn thng c ln da sm hn ngi dn thnh th? Ti sao ph n Vit Nam thch c ln da trng trong khi ph n u chu thch ln da bnh t? Ti sao cc nam phu thut vin hay chi th? Tn s chi th ca phu thut vin c khc nhau gia cc b mn? Ti sao bnh nhn t vong nhiu trong hai ngy cui tun? Cht lng cuc sng ca nhng bnh nhn sau khi x tr ra sao?

l nhng vn tuy n gin nhng i hi chng ta phi suy ngh, tm cu tr li, v trong vi trng hp, tm gii php. Nhng cu tr li c khi hnh thnh t cm nhn c nhn hoc lng nng bnh dn. Mt nh phu thut, qua kinh nghim lu nm, c th t tin rng phng php iu tr ca mnh l c hiu qu. Nhng cng c nhng cu tr li xut hin mt cch bt ng. Theo suy lun bnh thng, nu mt ngi hng xm mc bnh ung th v bc s cho bit s sng trong vng 3 thng nhng trong thc t sng n 3 nm sau khi dng mt loi tho dc, th ngi ta s ngh ngay n tho dc l c ch cho iu tr ung th. Nhng kinh nghim c nhn, nhng pht hin tnh
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c, tuy c th l chng c nhng cha phi l chng c khoa hc, v kh c th ng gp vo kho tng tri thc y hc, bi v cha c h thng ho. Mt cch h thng ho vn l qua nghin cu khoa hc. 1. Suy ngh nh nh khoa hc Do , ng trc mt vn , mt hin tng, chng ta phi tp cch suy ngh nh mt nh khoa hc. Nh khoa hc suy ngh c phn khc vi ngi thng, v h t khi no chu s chi phi ca cm tnh. Mt ca bnh cha thuyt phc h v mt liu php iu tr. C trng hp nhiu ca bnh cng cha thuyt phc, bi v h cn phi so snh vi nhm khng c iu tr (trong khoa hc, thng c gi l nhm chng control group). Ngoi ra, nh khoa hc cn phi pht biu gi thuyt (da trn cu hi) v kim nh gi thuyt qua th nghim. Trong trng hp ung th v dc tho, trc khi i n kt lun nh khoa hc phi tm hiu c ch sinh hc ca dc tho, t gi thuyt v hiu qu, v tin hnh nghin cu thu thp d liu xem c ph hp vi gi thuyt hay khng. Ni mt cch ngn gn, nh khoa hc suy ngh qua 3 bc: t cu hi, pht biu gi thuyt, v tin hnh th nghim. Cu hi nghin cu (research question) l mt pht biu mang tnh bt nh v mt vn . V mang tnh bt nh, nn nh khoa hc phi tm hiu nhng yu t no dn n s bt nh. Cn phi phn bit mt cu hi nghin cu tt vi mt cu hi d. Cu hi nghin cu tt phi ng ng t nht 3 trong 5 tiu chun m ti tm gi l FINER.

F l vit tt ca feasibility, tc tnh kh thi. Mt cu hi nghin cu tt phi kh thi, tc nh khoa hc c kinh nghim chuyn mn, c th tuyn dng y bnh nhn hay i tng nghin cu, c phng tin o lng v th nghim, v.v. I l interesting, tc th v. Mt cu hi nghin cu tt phi th v i vi nh khoa hc, xng ng theo ui. C nhiu nh khoa hc b ra c i ngi ch theo ui mt phn t rt nh. N l novelty, tc c ci mi. Lm nghin cu l mt vic lm sn sinh ra thng tin mi, phng php mi, hay tng mi. Mt nghin cu ch lp li y chang nhng g ngi khc lm th khng c ci g mi, v c xp vo nhm me too. E l ethics, tc o c. Mt nghin cu y khoa phi tn trng quyn con ngi, khng lm hi ngi, phi bo mt tuyt i (khng c tit l thng tin c nhn ra ngoi). Mt nghin cu y khoa cn phi p ng nhng tiu chun v y c. Nu khng p ng tiu chun o c / y c th d cu hi c th v c no cng phi b i! R l relevant, tc lin i. Tht ra, ch lin quan y c ngha l c nh hng. Mt cu hi nghin cu m nu tm c cu tr li v c th lm thay i chuyn ngnh l mt cu hi quan trng. Tc ng y phi hiu l c tc ng tch cc n thc hnh lm sng, n chnh sch y t ca Nh nc, hay c ng gp mt nh hng mi. Chng hn nh cu hi Cc chng trnh truy tm ung th v c hiu qu gim nguy c t vong? l mt cu hi tt v cu tr li s
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lm thay i nhn thc ca ph n v thay i chnh sch y t. Nhng cu hi nghin cu, m nu c cu tr li cng chng thay i g cho chuyn ngnh hay chng ng gp thm g v thng tin th khng ng theo ui. Gi thuyt i vi nh khoa hc rt khc vi cu hi nghin cu. Gi thuyt xut pht t cu hi nghin cu. Cu hi nghin cu xut pht t quan st thc t. Mt cu hi nghin cu tt s dn n gi thuyt khoa hc hay. Gi thuyt khoa hc l mt pht biu mang tnh tin lng gia hai hay nhiu bin. Hai ch tin lng y rt quan trng! Mt pht biu nh C mi lin h gia ung th v v long xng khng phi l gi thuyt v khng c tnh tin lng. Nhng mt pht biu nh Bnh nhn ung th v c nguy c long xng thp hn bnh khng b ung th v th c xem l mt gi thuyt khoa hc (v c tnh tin lng). Cng nn phn bit gi thuyt khoa hc vi gi thuyt thng k. Gi thuyt thng k ch l mt cch pht biu gi thuyt khoa hc tin cho vic kim nh thng k. Nu gi thuyt khoa hc l bnh nhn tiu ng c t trng m cao hn ngi khng b tiu ng, th gi thuyt thng k l bnh nhn tiu ng c t trng m bng ngi khng b tiu ng. Gi thuyt thng k va cp cn c khi gi l gi thuyt v hiu (null hypothesis). Bn cnh gi thuyt v hiu, nh khoa hc cn pht biu mt gi thuyt o (alternative hypothesis): bnh nhn tiu ng c t trng m khc vi ngi khng b tiu ng. Mt cch nghim tc, khng c gi thuyt v hiu th tt c phn tch thng k gn nh v ngha (bi v phn ln cc phng php thng k c pht trin ly gi thuyt v hiu lm chun). Mt trong nhng nhim v ca nh nghin cu l dng d liu bc b gi thuyt v hiu (v bc b gi thuyt v hiu l gin tip chp nhn gi thuyt o), bi v theo nh trit hc khoa hc Karl Popper, chng ta khng th no chng minh c mt gi thuyt khoa hc. Sau khi c gi thuyt, bc k tip l lm th nghim (experiment). Th nghim y nn hiu mt cch rng hn, ch khng n thun c ngha th nghim trong phng th nghim. Mt nghin cu lm sng i chng ngu nhin (randomized controlled trial RCT) m theo mt nhm bnh nhn c chia thnh hai nhm nh (nhm c iu tr v nhm chng khng c iu tr) c xem l mt th nghim th nghim lm sng. Mt nghin cu bnh chng cng l mt th nghim. Th nghim l mt bc cc k quan trng thu thp v phn tch d liu, i n kt lun lin quan n cu hi nghin cu. Do , c th ni rng nghin cu l mt chu trnh khp kn. Bt u t cu hi nghin cu, nh khoa hc pht biu gi thuyt, ri tin hnh th nghim kim nh gi thuyt . Mt cng trnh nghin cu tt thng m ra mt nh hng mi, v nhiu cu hi nghin cu mi. Chng hn nh cu hi Ti sao ngi nng thn c ln da sm hn ngi thnh th, sau khi c cu tr li (v d) l do vitamin D, th n s m ra hng lot nghin cu mi: ti sao ngi thnh th hay thiu vitamin D; yu t no quyt nh nng vitamin D trong mu; vitamin D nh hng n bnh no; nng vitamin D ti u l bao nhiu; v.v. Mt cng trnh nghin cu khng m ra mt hng mi l mt cng trnh khng xng ng theo ui.

2. cng nghin cu v vn phong Th nghim cn phi c qui trnh v k hoch. Nhng qui trnh v k hoch ny phi c hoch nh (trc khi tin hnh nghin cu) trong mt ti liu m ting Anh gi l Research Proposal, v dch sang ting Vit l cng nghin cu. Ti mun hiu hai ch cng nh sau: l ngh, xut; v cng l cng lnh. cng nghin cu, do , l mt ti liu m trong nh khoa hc ngh cng lnh hay chng trnh lm vic. y l ti liu quan trng nht trong mt cng trnh nghin cu, v qua m c quan ti tr c th xt duyt cp kinh ph. Ti ngh c th xem cng nghin cu nh mt bn v ca kin trc s. Tht vy, c th xem nh khoa hc nh l mt kin trc s. Kin trc s phc ho chi tit xy dng mt cng trnh trong bn v. Nh khoa hc phc ho nhng chi tit v qui trnh lm nghin cu thu thp v phn tch d liu. Nu bn v l mt tc phm ca kin trc s, th chng ta cng c th xem cng nghin cu nh l mt tc phm ch ngha khoa hc nhm i tm cu tr li cho cu hi nghin cu. l tc phm ch ngha, th cch vit cng nghin cu ng vai tr rt quan trng. Quan trng l v ngi c (c th l nh ti tr, hay hi ng xt duyt) da vo m quyt nh cung cp ti tr hay khng. Do , ngi vit (tc nh khoa hc) cn phi suy ngh rt cn thn, c h thng, v s dng ngn ng cho chnh xc. Ni th c l d, nhng thc hnh th khng d cht no. Ti liu ny c mc ch khim tn l chia s cng cc bn nhng kinh nghim v cch vit cng nghin cu y khoa sao cho thuyt phc v nng cao xc sut c ti tr. Vit cng nghin cu khc vi vit bi bo khoa hc. Trc y, ti vit mt lot bi v cch vit mt bi bo khoa hc (c th xem trang nguyenvantuan.net trong phn K nng khc). Nhng phng php vit bi bo khoa hc khng th p dng cho vit cng nghin cu. Nhng khc bit chnh l mc tiu v thi gian tnh. Mc tiu ca vit cng nghin cu l thuyt phc nh ti tr hay hi ng xt duyt rng chng ta c tng tt, c cch tip cn hay, c phng tin thc hin nghin cu. Mc tiu ca vit bi bo khoa hc l bo co nhng pht hin rt c th trong mt nghin cu v nhng pht hin ny c ngha g. V thi gian tnh, vit cng l phn nh v tng lai, cn vit bi bo khoa hc l bo co nhng g lm trong qu kh. Vit cng mang tnh ha hn, nhng ha hn mt cch khoa hc (tc c bng chng), do rt khc vi vit bo co khoa hc thng mang tnh bin minh v bin lun. Theo kinh nghim c nhn, ti thy vit cng nghin cu cung cp cho mnh nhiu c hi rt hay. Th nht l c hi sp xp tng mnh mt cch c h thng, c trc, c sau. Nhng tng hn n, khi c m t trn trang giy s lm cho chng ta suy ngh logic hn. Th hai, vit cng cng l mt c hi cp nht ho thng tin, v chng ta cn phi tm hiu trong y vn xem cc ng nghip khc lm g. Th ba, t , vit cng nghin cu cng c ngha l tm ng nghip mi. Nhn nhn nh th, vit cng c nhiu li ch, ch khng phi ch l v vi theo cch ni ma mai ca ngi ngoi cuc. Vit cng (hay vit vn ni chung) l mt cch suy ngh. C l chng ta u ng rng vit vn l mt phng tin chia s thng tin vi nhng ngi quan tm. Ngi
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quan tm c th l ng nghip ca mnh, nhng cng c th l ngi khng cng chuyn ngnh. Nhng ti mun ngh rng vit vn cng l mt cch thc hon thin tng, v hiu theo cch , vit chnh l mt phng tin hay mt cng c suy ngh. Suy ngh cng cn phi c phng tin, v theo ti vit xung nhng cu ch do mnh la chn chnh l mt phng tin. C l chnh v th m Nh vn [v cng l bc s] Verghese tng ni mt cu bt h, I write to understand what I think (ti vit hiu nhng g ti suy ngh). Ti rt ng vi cu ny. Th th cu hi t ra l tiu chun g nh gi cch vit cng nghin cu l tt hay d. Kinh nghim c nhn ti cho thy c 5 tiu chun: trong sng, n gin, chnh xc, khch quan, v cu trc logic. Trong sng c ngha l trnh nhng cu vn rm r, nhng t kh hiu. Nu vit Nng insulin nhm iu tr cao hn th s khng r rng, v ngi c khng bit cao hn nhm no. Ngay c cch vit Nng insulin nhm iu tr cao hn nhm chng cng c th ni l cha t, bi v cu vn hm ni rng tt c bnh nhn trong nhm iu tr u c nng insulin cao hn nhm chng mt tnh hung rt kh xy ra. Trong thc t th Tnh trung bnh, nng insulin nhm iu tr cao hn nhm chng c l r rng hn. Vn phong khoa hc rt khc vi th. Ti thng ni a [v hay ly lm v d] v cu th ca mt tc gi ti rt mn m (L t): Ti tha thn gia cha Qun Ng / Li chuc tui mnh ni tht khai sinh. Cu trch trong tp thng Ch. l mt cu vn rt kh hiu! Mt thi s khc ti cng rt ngng m l Hong Cm, ngi sng tc th mt cch siu thot. ng c sng tc bi Tnh Cm, trong c hai cu rt du dng: Nu anh cn tr nh nm c / Quyt n em v sng vi anh. C th ni l nhng cu th c o, gieo vn tuyt vi, m iu rt nhc tnh, nhng nh khoa hc khng th vit nh th c. Nh khoa hc phi vit vn trong sng. Chng hn nh nu ni cn tr th phi nh lng r rng bao nhiu tui l tr; nu ni nm c th phi ni nm no; nu ni v sng vi anh th phi ni sng u. Vn phong khoa hc khng th chung chung c. Nh khoa hc khng phi nh th. Nhng rt tic, nhiu khi ti bt gp nhng cu rt th trong nhng bi bo khoa hc t ng nghip Vit Nam. Mt trit gia c tng ph bnh ngi ng rng chng ta [ngi ng] hay ln ln gia th v khoa hc! Trong vn phong khoa hc, mt cu vn phi c thng tin. Cu vn khng c thng tin l cu vn tha. V d tiu biu cho cu vn khng c thng tin l [hay thy trong gii bo ch] Cng trnh nghin cu c 2 mc tiu. v c xong cu vn ngi c khng c bt c mt ni dung no c; ngi c k vng 2 mc tiu l g v t hi ti sao tc gi khng vit ra. Nhng nu vit Cng trnh nghin cu c 2 mc tiu: xc nh nh hng ca can thip, v xc nh yu t nguy c th l mt cu vn c thng tin. Th c 2 cu: Ngoi ba kha cnh trn, chng ti cn phn tch vn da trn l thuyt vn ho x hi. y, li c nhiu kha cnh khc.

Hai cu vn ny c th thch hp cho bo ch, nhng hon ton tht bi trong khoa hc! Ni cch khc, trong vn cnh khoa hc, mt cu vn phi tn ti mt cch ring l (self-contained), c thng tin v c ; c cu vn nh th ngi c khng hn cn phi c cu trc. n gin c ngha l dng t ng d hiu, chnh xc, v cu vn ngn. Trong ting Vit c nhng cu ch rt di m t mt , nhng nu c k c th vit ngn gn hn. Thay v dng nhng danh t gc Hn, chng ta nn c gng dng nhng danh t gc Vit. Chnh xc l nh lng ho ni dung thng tin. Trnh nhng t ng m m. Ting Vit ta (v ting Anh cng th) c nhng ch nh khong, xp x, , gn, a s, phn ln, ni chung, v.v. khng mang tnh nh lng cao. Khoa hc l cn o ong m, nn c gng vit mt cch nh lng. Kh bit bao nhiu l a s, 80% hay 90% l a s? Thay v vit a s bnh nhn , chng ta nn vit (nu c s liu) 80% bnh nhn th s r rng hn. Trong khoa hc khng c chuyn ni chung. Vit n y ti nh c ln ng Nguyn B Thanh cht vn mt gim c s giao thng vn ti thuc Thnh ph Nng, v ng gim c tr li Tha anh, ni chung l ., ng Thanh ngt li ngay: Ni ring, ch khng ni chung. Hi trng ci xo. ng Nguyn B Thanh p dng tiu chun khoa hc vy. Khch quan l cch vit phi cm tnh, v nht l khng nht ch vo ming ngi c. Thay v vit S khc bit rtc ngha lm sng, th nn vit S khc bit c ngha lm sng ri trch dn con s hay d liu ngi c phn xt. Khch quan cng c ngha l trnh gi nh (kiu nh Ai cng bit rng ). Tun theo nguyn tc khch quan cn c ngha l trnh nhng cu vn khng c chng c. Cu trc logic l phi c gng sp xp tng mt cch c trc c sau, khc chit. C mt thng k [ti khng cn gi ngun] cho rng 85% nhng hiu lm l do cu trc on vn, ch c 15% hiu lm l do ni dung. Do , c l y l tiu chun quan trng nht trong cch vit cng. Bt c khi nim g mi cn phi c gii thch trc . Nu mt on vn xut hin cm t cht lng cuc sng m khng c cp n trong cc on vn trc l mt cch vit rt d (thiu tnh khc chit). Cu trc cu vn v on vn nhn mnh mt ch . Chng ta th c 4 cu vn sau y: a. Mc d thuc c hiu qu cao, nhng cng gy tc hi ng k. b. Mc d thuc gy tc hi ng k, nhng c hiu qu cao. c. Thuc c tc hi ng k, nhng rt hiu qu. d. Thuc c hiu qu cao, nhng gy tc hi ng k Ci khc bit chnh gia cc cu vn c l khng phi ni dung, m l v tr nhn mnh tng. Cu hi t ra l trong mt cu vn, cn nhn mnh phn no: phn u, phn gia, hay phn cui? Cc chuyn gia v vit vn khuyn co rng nn dng phn u nhn mnh im chnh yu. (Cng c chuyn gia ngh nn dng phn cui cu vn nhn mnh). Nn m u cu vn vi mt t kho, nu chng ta mun ngi c ch n ch . Trong cu long xng l mt vn y t quan trng, ti nhn mnh n long xng. Tuy nhin, nu ti vit Mt trong nhng vn y t quan trng
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nht hin nay l long xng, th cng ni ln ci , nhng c l ngi c s hiu rng ti ang nhn mnh n kha cnh y t hn l long xng. V tr nhn mnh v cch dng c khi lm cho ngi c hiu khc. Trong mt cun sch v cch son cu hi trong nghin cu khoa hc, tc gi k chuyn [vui] v 2 thy tu, mt ngi tu dng a Minh (Dominican) v mt ngi tu dng Tn (Jesuit), tho lun xung quanh vn ti v ht thuc l trong khi cu nguyn. Sau mt lc tho lun hai ngi bt ng kin, v mi ngi i hi b trn ca mnh. Tun th hai, h gp li, v c cuc i thoi nh sau: V tu s dng a Minh hi v tu s dng Tn: Th b trn ca anh ni g? V tu s dng Tn: Ngi ni ok. Tu s dng a Minh: Vui nh! B trn ca ti ni rng l mt ti. Tu s dng Tn: Th anh hi b trn ca anh nh th no? Tu s dng a Minh: Ti hi ngi rng ht thuc l trong khi cu nguyn c ok khng? Tu s dng Tn ln ri ni: Ah, cn ti hi ngi rng cu nguyn trong khi ht thuc l c ok khng. l mt bi hc ng ch v cch t ch vo v tr thch hp. Ch cn thay i th t ca ch l c th thay i ngha ca cu vn, v lm cho ngi c c th hiu khc. Trong vit vn khoa hc, tc gi khng nn cho ngi c hiu hai ngha! Ngoi ra, nn trnh cch vit nh trng b di. l cch vit tuyn b mt ch , nhng nhng cu sau li ni v ch khc. Th c on vn: Gy c xng i l hu qu hay gp ng hng th 3 ca long xng, bnh c t l t vong v tn ph cao, cht lng sng ca ngi bnh gim ng k ngay c khi c iu tr ng n. Bnh thng gp ngi cao tui vi t l n:nam l 3:1. Cu u ca on vn trch dn trn c 3 chnh: s ph bin, h qu, v iu tr. c xong cu vn , ngi c k vng tc gi s chng minh ba bng d liu ra sao. Nhng cu th hai th tc gi khng cung cp thm thng tin b ngha cho cu u, m chuyn sang mt khc. i vi tc gi c kinh nghim, on vn trn cha t. l cha ni n cch dng ch v ngha cn kh m m (v d: ng hng th 3 ca long xng c ngha l g). Trong vn phong khoa hc, mi cu vn ch ni ln 1 m thi; tham vng qu s khng chuyn ti ht ngha.

4. Ni dung mt cng nghin cu Mt cng nghin cu bao gm nhng g? Tr li cu hi ny cn tu thuc vo vn ho v qui nh a phng. Vit Nam, nhng cng nghin cu thng c nhng mc sau y:

Mc tiu ca ti; Tng quan tnh hnh nghin cu v tnh cp thit ca ti ; Ni dung nghin cu; Phng php nghin cu, cch tip cn vn ; Hp tc quc t (nu c) ; Tin thc hin;

Dng kt qu d kin ca ti; Yu cu khoa hc i vi nhng sn phm d kin to ra; K hoch trin khai; Li ch mang li v tc ng ca kt qu nghin cu; Cc t chc tham gia nghin cu; v Kinh ph. Nhng mc ca mt cng nghin cu trn y khng c vn g, nhng cch t chc v sp xp thng tin th hi khc so vi nc ngoi. Nhng mc nh cc t chc tham gia nghin cu, kinh ph l vn hnh chnh, ng l phi trnh by trong mt ti liu ring. nc ngoi (c th l c, Anh, c, Thi Lan, M, v.v., nhng ni m ti c kinh nghim) th ni dung mt cng nghin cu thng n gin hn Vit Nam. nhng nc , mt cng nghin cu bao gm nhng phn sau y:

Tm lc (synopsis): ging nh mt tm tt cho mt d n nghin cu. Thng thng ch 1-2 trang giy; Mc tiu c th (specific aims); Bi cnh v tm quan trng (background & significance); Kt qu nghin cu s khi (preliminary results);
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K hoch nghin cu v phng php (research design and methods); v Ti liu tham kho.

Tuy nhng mc ca cng Vit Nam c v hi nhiu, nhng s trang thng ch dao ng trong khong 10 n 20 trang. nc ngoi, nh NIH (Vin Y t ca M chuyn cp ti tr cho nghin cu y sinh hc v cng l mt trung tm nghin cu) th c qui nh s trang rt r rng. Mt cng ca NIH ti a l 30 trang (hin nay th c thay i v gim xung cn 20 trang?) Ti c lm mt so snh gia mt cng tiu biu ca VN v ca NIH th thy nh sau: So snh s trang ca mt cng VN v NIH mc Tng liu quan Vit Nam M (NIH) ti 10 1 2 1 6-8 10-20

Mc tiu

Kt qu s khi 0 Phng php 5

Ni cch khc, cng nghin cu ca VN vit di v bi cnh v tng quan ti liu, nhng rt ngn v phng php. Ngc li, nc ngoi (tiu biu l NIH ca M), mt cng nghin cu ch yu l phn phng php (rt di) nhng phn tng quan ti liu th ngn. Rt nhiu cng nghin cu t VN m ti c qua, trong phn tng quan ti liu, tc gi vit di nhng chng lin quan g n ch v mc tiu nghin cu! Hu nh cng nghin cu y khoa no cng vit theo mt cng thc, trong c c phc iu tr, nhng nh ngha rt cn bn v bnh, nhng yu t nguy c (m c l ai trong ngnh cng bit). C khi cng nghin cu tm hiu t l mc bnh, nhng tc gi phi im qua mt cch kh di dng v phng php iu tr! Phn ln nhng thng tin ny tht ra l tc gi dch t sch gio khoa, hoc dch t nhng bi tng quan trn cc tp san nc ngoi (v khng bun sa biu ting Anh hay ghi ngun!) ch tc gi cng cha hn am hiu. Mt im khc bit ng ch l cng nghin cu nc ngoi phi c phn Kt qu s khi, cn Vit Nam th khng c hoc khng yu cu. c v M, mt cng m khng c kt qu s khi (hay nhng nghin cu trc y cng ch m nh khoa hc lm) th khng bao gi c qua vng u xt duyt, rt rt kh c kh nng c ti tr. Trong cc phn sau y, ti s trnh by cch vit 4 phn quan trng nht: mc tiu, bi cnh, nghin cu s khi, v phng php nghin cu. Ti s c gng a vi v d c th t nhng cng trc y ca ti v ca cc ng nghip khc. V bi ny cng nhm n cc bn nghin cu sinh ang hay sp i hc nc ngoi, nn ti cng trnh by vi v d bng ting Anh cc bn c th tham kho. 4.1 Phn mc tiu nghin cu

Mc tiu nghin cu l phn quan trng ca mt cng nghin cu, v l b mt m ngi c s nhn qua. Khi ngi c thy mc tiu nghin cu c ci g mi hay th v th h s c tip; nu khng, h c th xp li cng v th l tc gi tht bi. Do , c th xem phn mc tiu nghin cu nh l mt ci test cho ngi c. Ch khi no ci test ny c qua th tc gi mi thnh cng mt phn trong vic thuyt phc ngi c. Phn mc tiu nghin cu nn c cu trc 3 phn nh sau: Mt on vn tng qut v vn chung; Pht biu v mc tiu chung (ting Anh gi l goal); Pht biu v mc tiu chuyn bit (specific aims), v mi mc tiu nn km theo mt gi thuyt. on vn tng qut phi ni c vn ln m nh nghin cu quan tm. l mt on vn mang tnh holistic, hay bi cnh nghin cu m cng ny s ng gp mt phn tri thc.

Mc tiu chung hay mc tiu tng qut l mc tiu lu di m nh nghin cu mun t c. C th nghin cu hin nay cha t c, nhng s ng gp mt phn trong nh hng gii quyt vn v lu v di. Trong bng , mc tiu ti hu v tng qut l ph li; tng t, trong nghin cu khoa hc, mc tiu tng qut l nhm n gii quyt mt vn chung cho chuyn ngnh. Mc tiu chuyn bit l nhng tm im c th. C th hiu theo ngha c th o lng c, c th kim nh c. Ngi vit cng kinh nghim thng vit mi mc tiu chuyn bit km theo mt gi thuyt. Thnh thong, nu c t, nh nghin cu nn km theo nhng kt qu d kin cho tng mc tiu. Thng thng ch cn 2-3 mc tiu chuyn bit l . Nhiu mc tiu qu s kh thc hin; t qu s khng p ng tiu chun ca mt d n nghin cu khoa hc. Trong thc t, ti tng thy nhiu cng tht bi (k c ca chnh ti) v cch pht biu mc tiu cha t. Nhng sai lm ph bin trong cch vit mc tiu chuyn bit bao gm:

Mc tiu phi thc t, tc thiu tnh kh thi; Mc tiu khng c l do chnh ng (ging nh t trn tri ri xung); Mc tiu mang tnh qu m t (nh tm hiu bao nhiu nam v n c hi chng chuyn ho); Mc tiu qu phc tp, m c ln ngi ta khng hiu tc gi mun lm g; Mc tiu qu ph thuc ln nhau (s bn di y).

V d 1: y l trch dn cng nghin cu v long xng ca mt ng nghip v c s ng ca tc gi. [on vn tng qut du dt c gi rng y l vn quan trng]: Long xng l mt bnh m lc ca xng b suy gim, cht lng xng xung cp, v h qu l
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tng nguy c gy xng. H qu ca long xng l gy xng. Chn on long xng da vo o lng mt xng. Tuy nhin, Vit Nam cha c gi tr tham chiu cho vic chn on long xng. [Mc tiu tng qut p ng vn chung]: Mc tiu lu di ca nghin cu ny l pht trin tiu chun chn on long xng cho ngi Vit (nam v n) v xy dng m hnh tin lng gy xng. Hon thnh mc tiu ny s gip cho vic chn on long xng chnh xc hn, v nhn dng nhng c nhn c nguy c gy xng cao can thip sm v gim qui m long xng trong cng ng. [Mc tiu chuyn bit] t c mc tiu , chng ti ngh theo ui cc mc tiu c th nh sau:

Mc tiu 1: Xy dng gi tr tham chiu mt xng i v xng ct sng nam v n; v qua , xc nh ch s T cho chn on long xng ngi Vit. Gi thuyt 1: Mt xng ngi Vit thp hn mt xng ngi u M; do , ch s T ca ngi Vit cng khc vi ch s T ca T chc Y t Th gii. Kt qu k vng: Gi tr tham chiu v mt xng (trung bnh, lch chun, ch s T) c th s dng cho vic chn on long xng nam v n Vit Nam. Mc tiu 2: Xc nh t l lu hnh long xng v gy xng nam v n; Gi thuyt 2: N gii c nguy c long xng v gy xng cao hn nam gii, nhng khc bit s gim dn trong tui trn 70. Kt qu k vng: Kt qu l tn s long xng v gy xng theo gii v tui. Nhng kt qu ny s cung cp chng c v qui m long xng v gy xng ngi cao tui v sau mn kinh (n).

Mc tiu 3: Xc nh yu t nguy c lin quan n long xng v gy xng, k c cc markers chu chuyn xng, v qua pht trin m hnh tin lng long xng v gy xng. Gi thuyt 3: Marker chu chuyn xng tng theo tui, v chu chuyn xng cao tng quan vi s suy gim mt xng nam v n. Kt qu k vng: Gi tr tham chiu v marker chu chuyn xng, v m hnh tin lng gy xng cho ngi Vit. Vi m hnh ny, cc bc s c th c tnh nguy c gy xng ngay c trong iu kin khng c my DXA. V d 2: Trong v d ny, tc gi vit phn mc tiu rt bi bn.

[M u l vn chung]. Exposure to pathogenic microbial lipids, like lipopolysaccharide (LPS), triggers a complex and coordinated protective response by the immune system. A growing body of evidence indicates that triglyceride-rich lipoproteins and apolipoprotein E (apoE) play an integral role in host defense against bacterial infection. Yet, how these non- traditional elements of the immune system contribute to host immunocompetence is unclear. Published data indicate that apoE is protective against bacterial infection and injury. Accordingly, infusion of apoE has been shown to decrease LPS-induced morbidity and mortality in rodents. [2, 3] Also, apoE-deficient mice have an increased susceptibility to lethal infection when injected with live bacteria.
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[4, 5] But, unexpectedly, our laboratory has recently discovered that infusion of apoE increased rather than decreased mortality after cecal ligation and puncture, an in vivomodel of polymicrobial sepsis. [1] We believe that this discordant observation highlights a novel activity for apoE in regulating the host response to pathogenic microbial lipid antigens through activation of thymus-derived lymphocytes (T cells). Consequently, uncovering the role that triglyceride-rich lipoproteins, apoE and T cells play in the mammalian response to infection simultaneously assigns important new biological functions to plasma lipoproteins, further blurs the boundary separating innate and adaptive immunity, and provides unique insights into the host response to infection that could yield innovative therapies for sepsis. [dng gch dng nhn mnh mc tiu chung] This proposal will investigate how apoE and natural killer T (NKT) cells, a sub-population of T lymphocytes, contribute to the host response to severe sepsis following cecal ligation and puncture in mice. Furthermore, we will test the hypothesis that modifying the expression or activity of apoE can protect against sepsis. [Pht biu gi thuyt chung] Our working hypothesis that triglyceride-rich lipoproteins are integral components of the immune system is supported by the following observations. First, the synthesis and secretion of triglyceride-rich lipoproteins is dramatically increased during clinically significant infections, an observation termed lipemia of sepsis. Second, triglyceride-rich lipoproteins bind various microbial lipids and thus protect against shock and death in rodent models of sepsis. [6-9] Third, triglyceride-rich lipoproteins clear LPS from the circulation and deliver it to the liver [7, 10], where lipoprotein-LPS complexes subsequently modulate the hepatic immune response to infection. [11-14] And, fourth, apoE has recently been shown to bind and deliver microbial lipids to antigen-presenting cells, a critical step in activating NKT cells and the immune system. [Pht biu gi thuyt chuyn bit] The specific hypothesis driving the proposed research is that apoE, a key constituent of triglyceride- rich lipoproteins, regulates the host response to severe infection through its effects on NKT cell activation and cytokine production. By examining the effect of apoE on an in vivo model of polymicrobial sepsis in mice, we aim to uncover the regulatory impact of apoE on the immune response to infection.Our long term goal is to identify how plasma lipoproteins contribute to host immunocompetence and apply this knowledge to the development of novel and effective treatments for severe bacterial infections. [Mc tiu chuyn bit, dng t m nhn mnh v gy ch . Trong mi mc tiu, tc gi ch ra h s c kt qu g] The specific aims of the proposal are therefore: 1. To demonstrate that serum apolipoprotein E (apoE) concentrations correlate with morbidity and mortality in a murine model of polymicrobial sepsis. A. show that apoE increases mortality following cecal ligation and puncture (CLP) sepsis in mice in a dose-dependent manner; B. show that apoE increases CLP-induced morbidity via changes in Th1 cytokine secretion, liver injury and bacterial clearance;

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2. To demonstrate that apoE promotes the activation of natural killer T (NKT) cells during CLP-induced sepsis. A. delineate the effect of apoE on NKT cell frequency, proliferation, cytokine expression and cytotoxic effector functions in the liver, spleen and thymus following CLP in mice; B. show that apoE-mediated immune regulation during sepsis is dependent on NKT cell activation using immunodeficient mice; 3. To test the hypothesis that inhibition of apoE activity protects against the morbidity and mortality of sepsis. A. show that the biochemical, immunologic or genetic inhibition of apoE activity protects against sepsis; B. examine the effect of modifying apoE activity during the early versus late phase of sepsis. V d 3: y l mt cng nghin cu lm sng M. Tc gi c cch vit c ng hn. M u, tc gi m t mt cch ngn gn v ni dung ca nghin cu. Sau , tc gi pht biu mc tiu chnh (primary aim) v mc tiu ph (secondary aim). [M u, m t ngn gn v cng trnh IRIS v gi thuyt] The Insulin Resistance Intervention after Stroke Trial (IRIS) is a randomized, double-blind, placebo-controlled trial that will test the hypothesis that reducing insulin resistance and its sequelae with thiazolidinedione therapy will prevent stroke and myocardial infarctionamong patients with a recent ischemic stroke. Eligible subjects are men and women over 44 years of age without diabetes mellitis who have insulin resistance and a non- disabling ischemic stroke. During 3 years of recruitment, 3136 patients will be randomly assigned to pioglitazone, a thiazolidinectone, or placebo. [Mc tiu chuyn bit] The specific aims are as follows. 1. Primary Aim. To determine if pioglitazone, compared to placebo, will reduce the overall risk for fatal or non-fatal stroke or fatal or non-fatal MI among non-diabetic men and women over age 44 years with insulin resistance and a recent ischemic stroke. Among diabetics with insulin resistance, we hypothesize that pioglitazone will reduce the occurrence of any primary endpoint (fatal or non-fatal stroke or MI) within four years from 27% to 22%. The basis of this hypothesis is research showing... (details about studies associating insulin resistance with increased risk for stroke, MI, etc.)...By these and other mechanisms, we hypothesize that pioglitazone will protect patients with ischemic stroke and insulin resistance against recurrent vascular events.

Trong cc v d trn y, d dng thy rng cc mc tiu chuyn bit tuy nhm n mt mc tiu tng qut, nhng khng l thuc vo nhau. y l mt chin lc trong vic son cng nghin cu. Nu mc tiu 2 tu thuc vo s thnh bi ca mc tiu 1, v mc tiu 3 ph thuc vo kt qu ca mc tiu 2, th n s rt nguy him. Nguy
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him l v nu mc tiu trc tht bi, th tt c cc mc tiu khc u khng th thnh hin thc, v do , cng trnh nghin cu s khng kh thi. Bi hc y l cn phi thit k nghin cu sau cho cc mc tiu chuyn bit t ph thuc vo nhau.

4.2 Phn bi cnh v tm quan trng Phn bi cnh v tm quan trng, nh tn gi, c chc nng dn dng sn khu cho cng trnh nghin cu. Khng ch dn dng, m cn phi ni ln c tm quan trng ca cng trnh nghin cu. Khng ai mun u t vo nhng cng trnh nghin cu khng gy nh hng hay tc ng tch cc. C 2 phn chnh l bi cnh (background) v tm quan trng (significance): Trong phn bi cnh, tc gi cn phi vit theo phong cch k chuyn. Nhng k chuyn mt cch khoa hc, sao cho ng nghip khng cng chuyn ngnh vn c th hiu c vn . Mi mt mc tiu chuyn bit trong phn u cn phi c l gii trong phn bi cnh. Phi vit v thuyt phc ngi c s c tip, v mun t mc ch , tc gi cn phi vit theo cng thc chung: Ci g bit --> ci g cha bit --> cu hi Known --> unknown --> question

Ni cch khc, tc gi phi im qua nhng nghin cu trc ( bit), v ch ra cho c khong trng tri thc (cha bit), t thnh cu hi cho nghin cu. V d chng ta bit tnh trng thiu vitamin D cc tnh min Nam, nhng v min Bc c thi tit kh hu khc v cha ai bit tn s thiu vitamin D ngoi Bc, nn cu hi l t l thiu ht vitamin D c dn pha Bc l bao nhiu. V cch vit, khng ch n gin lit k nhng nghin cu trc, m phi dng phng php 4C. Phng php ny vit tt t 4 ng t:

Compare so snh: tc gi cn phi so snh nhng thng tin t nhng nghin cu trc; Contrast i chiu: sau i chiu v gii thch ti sao c s khc bit; Cite trch dn: iu bt buc l nu dng d liu ca ng nghip th phi trch dn; v Critique ph bnh: Ph bnh y c ngha l ph bnh mt cch knh trng, ch khng mang tnh nh ng nghip. Cch vit hay nht l thay v ph phn, tc gi c th trnh by mt cch hiu, cch din gii khc xem nh l mt cch ng gp vo y vn.

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Trong phn tm quan trng (significance), tc gi cn phi gii trnh rng cng trnh nghin cu s c tc ng n: Chuyn ngnh; Chnh sch y t hay thc hnh lm sng; Phng php mi; v Tri thc s rt ra c t cng trnh nghin cu. Phn tm quan trng c khi phi dng n k thut pitch (ln ging). Ni cch khc, cn vit sao cho ngi khc c th trch dn mt cu t cng. Chng hn nh nu l nghin cu v di truyn, ti c th vit

The studies in this proposal will provide a basis for understanding allelic heterogeneity influencing clinical endpoints, ultimately impacting on disease development (nhng nghin cu m t trong cng ny s cung cp nn tng hiu bit v s a dng alen c nh hng n kt cc lm sng, v sau cng l tc ng n s tin trin ca bnh). Mt cu nh th i vi vi ng nghip Vit Nam s gi l n, nhng nc ngoi th hon ton c th chp nhn c. Vn cn tu thuc vo v tr ca tc gi v uy tn trong chuyn ngnh. Nghin cu sinh c l khng nn vit khim tn hn, nhng vi ngi c tn tui th mt cu pitch nh th hon ton bnh thng. Nn nh rng vit cng l mt cch bn tng, nn tc gi cn phi thuyt phc tm quan trng ca nghin cu. V d 4: Trong cng cu di y, tc gi mun thuyt phc ngi c v tm quan trng ca cng trnh nghin cu: Long xng v gy xng l mt vn y t cng ng ln nc ta, v hng nm c khong 200.000 ngi gy xng, dn n gim tui th v hn ch lao ng. Mt xng l mt ch s lm sng quan trng v MX c th tin lng nguy c gy xng cho mt c nhn. V th, MX cn c s dng chn on long xng. Cng trnh nghin cu ny c mc tiu xy dng gi tr tham chiu MX cho ph n v n ng Vit Nam. Vi gi tr tham chiu ny, vic chn on long xng ngi Vit s chnh xc hn v qua chng ta c th bit c qui m long xng nc ta. Do , cng trnh nghin cu mang tnh cp thit, v s hin din ca my DXA nhiu nc ta nhng cha c gi tr tham chiu cho ngi Vit. V th, kt qu nghin cu c gi tr thc tin, c th p dng ngay cho vic chn on long xng. ngha l lun ca cng trnh nghin cu l cung cp nhng thng tin khoa hc cho vic hoch nh cc chin lc phng chng bnh long xng qui m cng ng.

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V d 5: Trong cng di y, tc gi vit ngn gn v bi cnh v tm quan trng nghin cu, bng cch dng cc tiu nh nhn mnh tng im: Why Study Square Cell Disease in the Kidney? Renal dysfunction commonly complicates square cell disease and is a major cause of morbidity and mortality. Acute Renal Syndrome is the leading cause of death in square cell disease and commonly leads to acute renal failure (58), while chronic uremia, filtration insufficiency, and renal vascular disease occur in 20-60% of adults with square cell disease (46, 54). Despite its clinical importance, the kidney has rarely been the focus of basic research in square cell disease. Current understanding of square cell pathophysiology derives from studies performed in other organs or in vitro. Because mechanisms of vaso-occlusion and inflammation in the kidney are likely to be different from those in other organs, there is a critical need for basic research on square cell disease that focuses on the kidney. Physiological Determinants of Vaso-occlusion in the Kidney Research on renal vaso-occlusion is limited to two studies that suggest severe medullary hypertonia causes sequestration of SQ RBCs in the kidney (3, 17). These studies did not adequately assess effects of modest tubular hypertonia and no study has evaluated the importance of mixed arterial hypertonia or inflammation to renal vaso-occlusion. In Specific Aim 1, we adapt the isolated rat kidney model used in these original studies (3, 17) to determine effects of tubular hypertonia, and mixed arteriolar hypertonia and renal inflammation on kidney micro vaso-occlusion...etc. Summary and Clinical Significance Studies performed in vitro and in other organs have given important insights into the pathophysiology of square cell disease, but have not yet defined the important pathophysiology in the kidney. The studies we propose will attack the problem directly using sensitive and specific techniques. These studies will lay the experimental foundation for understanding square cell disease crises in the kidney. The importance of these studies to the affected population cannot be exaggerated.

4.3 Phn nghin cu s khi

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 cng nghin cu Vit Nam thng khng c phn ny! Nhng i vi cc c quan ti tr nghin cu khoa hc nc ngoi th y l phn khng th thiu c. Tuy ni l nghin cu s khi, nhng trong thc t th nhng nghin cu nh th cng b trn cc tp san quc t. y l nhng nghin cu lm nn tng tc gi lm c s cho cu hi nghin cu v pht biu gi thuyt. Phn kt qu s khi cn c mc ch quan trng khc l thuyt phc ngi c rng tc gi c kinh nghim. Qua phn nghin cu s khi, ngi c c th nh gi tc gi hay nhm nghin cu c sn k thut, phng php, hay cng ngh cn thit thc hin cng trnh nghin cu. C l quan trng hn l c quan ti tr cm thy thuyt phc rng tc gi c th thc hin nghin cu (h an tm chn mt gi vng!) Thng thng, phn ny chim khong 6-8 trang giy, v nh ni trn, tc gi c th a vo nhng cng trnh nghin cu lin quan cng b trc y. V d 5: Mt cch trnh by hu hiu phn kt qu s khi l trnh by theo tng mc tiu chuyn bit. Trong v d di y, tc gi trnh by Preliminary Data This proposal is a collaboration between the PI and Dr. Barry Hurlburt in the Department of Biochemistry and Molecular Biology. The collaboration takes advantage of the expertise of the PI in the molecular genetics of S. aureus and the biochemical expertise of Dr. Hurlburt in transcription factor structure and function (14,15). The overall goals are 1) correlation of the expression of the sarA, sarB and sarC transcripts with the production and activity of SarA, 2) characterization of the mechanism by which sar regulates expression of the S. aureus collagen adhesin gene (cna) and 3) identification and characterization of additional S. aureus genes under the direct regulatory control of SarA. We have assembled all of the experimental tools required to accomplish these objectives. Specifically, we have (i) purified SarA in a form capable of binding an appropriate DNA target, (ii) generated an affinity-purified antibody against purified SarA, (iii) constructed a xylE reporter plasmid that can be used to assess the functional activity of SarA (Specific Aim #1) and define the sequence characteristics required for the regulation of cna transcription (Specific Aim #2), (iv) cloned the regions encoding the sarA, sarB and sarC transcripts for use in complementation experiments, (v) demonstrated that SarA binds a DNA target upstream of cna and begun the process of localizing the SarA binding site and (vi) obtained or generated sar and agr mutants in both cna-positive and cna-negative S. aureus strains. The experiments done to accomplish each of these tasks are described in detail below. Preliminary data for Aim #1. Cloning and expression of sarA. The polymerase chain reaction (PCR) was used to amplify the sarA coding region from S. aureus strain RN6390. Utilizing NdeI and BamHI restriction sites incorporated into the oligonucleotide primers, the fragment containing the sarA coding region was cloned into the E. coli expression vector pET9A. Because the NdeI site (CATATG) in the vector overlaps an ATG start codon, cloning of the sarA coding region into the NdeI site places the sarA structural gene in perfect register with the vector-derived ribosome binding site. Recombinant proteins are therefore expressed as full-length, wild type proteins without fusions to exogenous peptide or protein tags. After cloning the sarA PCR fragment into
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pET9A and confirming the identity of the cloned fragment by DNA sequencing (data not shown), the recombinant plasmid (pETSarA) was used to transform E. coli strain BL21(DE3)pLysS. Transformants were grown to mid-log phase before inducing SarA expression by adding IPTG to a final concentration of 0.4 mM. After two hours, cells were harvested and lysed by sonication. The presence of SarA in the crude lysate was confirmed by SDS-PAGE followed by Coomassie Brilliant Blue staining (Fig. 7). Preliminary data for Aim #2. Purification of SarA. A 500 ml culture of the BL21(DE3)pLysS E. coli strain containing pETSarA was induced and lysed as described above. After removing the insoluble material in the crude lysate by centrifugation, the soluble fraction was subjected to a series of ammonium sulfate precipitations culminating at 70% saturation. The pellet from each precipitation was resuspended in SDS-PAGE buffer and examined along with an aliquot of the supernatant (Fig. 8, left). The supernatant remaining after the final precipitation was found to contain ~70% SarA.

4.4 Phn phng php (reseach approach) Sau phn Mc tiu, Bi cnh & tm quan trng, Nghin cu s khi, l phn Phng php. y l phn di nht v chi tit nht so vi 2 phn trc. Mc ch ca phn Phng php l thuyt phc ngi c rng nh nghin cu: C k hoch tt kim nh gi thuyt t ra trong phn Mc tiu; C kin thc, k nng, v phng tin thc hin cng trnh nghin cu; ngh n nhng tnh hung xu s gp phi v c k hoch i ph; v Din gii kt qu d kin mt cch khch quan. Nn nh rng trong phn phng php, tc gi phi d kin tnh hung bt li s xy ra trong khi thc hin nghin cu. Khng mt nghin cu no u c tin hnh mt cch thun bm xui gi c. Bt c nghin cu no cng c vi trc trc, khng ln th nh, v c th nh hng n vic thc hin cc mc tiu. Chng hn nh trong nghin cu v long xng, c th nh nghin cu s gp kh khn nu my DXA h

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hng, hoc bnh vin thay i k thut vin, hoc cc mu sinh phm b nhim gy kh khn cho phn tch sinh ho, hoc cc bnh nhn t chi tham gia, v.v. Tt c nhng tnh hung ny phi c ch n trc khi tin hnh nghin cu. Do , nh nghin cu c kinh nghim phi suy ngh n tnh hung xu v c k hoch i ph. Cch vit hiu qu nht cho phn phng php l vit cho tng mc tiu. Nu cng c 3 mc tiu chuyn bit, phn phng php phi c 3 phng php tng thch. Cu trc phn phng php c th l: D. K hoch th nghim D.1 K hoch cho mc tiu 1 D.1.1 Thit k, l do, tm quan trng D.1.2 Phng php cho mc tiu 1 D.1.2.1 Ci mi D.1.2.2 Hn ch D.1.2.3 Kh khn c th tin on trc D.1.2.4 K hoch khc phc D.1.2.5 H qu D.1.3 Phn tch d liu D.1.4 Din gii kt qu tin on trc D.2 K hoch cho mc tiu 2 [] D.3 K hoch cho mc tiu 3 [] V d 6: Trong cng di y, tc gi m t phn phng php c th ni l rt y . Ngi c gn nh bit chnh xc tng bc tc gi sp lm g, ti sao lm, lm nh th no, v kt qu d kin ra sao.

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Research Design and Methods: The first specific aim of this grant application is to use the HFIM system to show that monotherapy with amantadine or oseltamivir carboxylate will lead to the emergence of resistance in influenza virusinfected cells and to demonstrate that the resistant viruses produced in the HFIM system under these conditions have the same mutations as those that emerge when people are treated with these drugs. In the second specific aim, we will use the HFIM system to optimize the dose and schedule of administration of current antiviral compounds effective against influenza viruses, delivered as monotherapy, to minimize the emergence of resistance. Finally, in the third specific aim we will determine the optimal dose and administration schedule of these anti-influenza virus drugs administered in combination therapy to prevent virus infection and the emergence of resistance. Specific Aim #1. Validate the HFIM as a model experimental system for influenza virus infection and the generation of drug resistant mutants. A. Introduction. Treatment of patients infected with type A influenza viruses with amantadine/rimantadine is known to lead to the rapid emergence of resistant viruses in the treated population (1-3). Treatment of patients with influenza with the neuraminidase inhibitors, oseltamivir carboxylate or zanamivir, usually does not lead to the emergence of resistant viruses (48). However, recent data have shown that treatment of children with influenza with oseltamivir carboxylate has led to the emergence of neuraminidase inhibitor-resistant influenza viruses (4-6). Data presented in the preliminary results section of this grant application showed that treatment of MDCK cells infected with a clinical isolate of influenza A virus in the HFIM system with amantadine can lead to the emergence of resistant viruses within two to three days of initiation of treatment. Phenotypic, but not genotypic, resistance was demonstrated when influenza virus-infected MDCK cells were treated with the D-tartrate salt of oseltamivir carboxylate in the HFIM system. The purpose of this portion of the grant application is to confirm these observations with A/Albany/1/98 influenza virus and to expand that observation for amantadine to additional influenza A viruses and for oseltamivir carboxylate to additional influenza A and B viruses. B. Experimental Design. We will examine the effect of amantadine and oseltamivir carboxylate on the replication of wild type rgA/Vietnam/1203/2004xA/PR/8/34 (a surrogate for avian H5N1 influenza virus), A/Texas/36/91(H1N1), A/Sydney/5/97(H3N2), and A/Victoria/3/75(H3N2) in the HFIM system. For comparison, we will also include our original clinical isolate, A/Albany/1/98(H3N2), to be certain that our original observations are reproducible for amantadine and oseltamivir carboxylate. Oseltamivir carboxylate will be tested against B/Lee/40 and B/Memphis/20/96 viruses. [] C. Expected results. Resistance will emerge under monotherapy. Amantadine resistant strains will have mutations in the M2 gene (residues 26, 27, 30, 31); neuraminidase inhibitor resistant strains will have mutations in the NA gene (residues 274 and 292) and/or HA genes (multiple residues). D. Potential problems. It is often difficult to generate mutations in vitro in the
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neuraminidase genes in the presence of neuraminidase inhibitors that resemble the mutations identified in the clinic. This may be due to the use of MDCK cells which have inappropriate cell surface receptors for influenza viruses. To address this potential problem, we will use a variety of other cell lines which more closely reflect the surface characteristic of lung epithelial cells such as A549 pulmonary alveolar epithelial cells (82), St Jude porcine lung (SJPL) cells (83), ST6Gal I cells (84) or SIATI cells (85) which express cell surface receptors with more terminal sialic acid, and Mink lung cells (86) to perform these dose ranging studies aimed at producing resistant viruses in the HFIM system. It is expected that by using the appropriate cell lines, resistant strains will be produced that more accurately reflect the neuraminidase inhibitor-resistant strains that have been identified in the clinic. E. Time frame. If this grant application is funded we will be able to purchase 4 additional duet pumps for the hollow fiber experiments thus doubling our capacity to perform these experiments. We plan to perform dose ranging experiments for amantadine and oseltamivir carboxylate on A/Victoria/3/75, A/Texas/36/91, rgA/Vietnam/1203/2004xA/PR/8/34, and A/Albany/1/98 and oseltamivir carboxylate for B/Lee/40 and B/Memphis/20/96 in the HFIM system. Each experiment will be repeated at least 1 time. One hollow fiber experiment takes approximately two weeks to perform from setup to take down. Analysis of virus yield (plaque assay, TCID50 assay and real time quantitative PCR) will take an additional two weeks. Therefore, each experiment, including a repeat, will take approximately 2 months. We plan to study at least the four type A and two type B viruses listed above for two drugs for a total of 24 hollow fiber experiments. Since we can study two viruses at a time for one drug or one virus for two drugs, Specific Aim 1 will take at least one year to complete.

cng nghin cu c vit cho i tng l ng nghip, nhng l ngi ng vai tr bnh duyt. Cu hi t ra l ngi bnh duyt k vng g khi c mt cng nghin cu. Bit c k vng ca h cng l bit cch p ng. Ti t mnh vo vai tr ngi duyt cng, v ti s k vng nhng iu qua nhng cu hi sau y m ti mun tm cu hi:

tng th v, cch tn, c th ng gp cho chuyn ngnh hay khng? D liu s khi c mnh hay thuyt phc tc gi tin hnh nghin cu ny? Cch tip cn vn ca tc gi c kh thi khng?

Chng c v kh nng v thnh tu ca tc gi ra sao? Trong yu t ny, ti mun xem qua thnh tch trong thi gian 5 nm gn y; cng c son mt cch r rng, logic, v chi tit hay khng? Cch vit trong sng v gn (v iu ny phn nh t duy ca tc gi). Nghin cu khoa hc i hi suy ngh v tnh t m. Khoa hc khng chp nhn suy ngh hi ht. Nhng suy ngh m m (muddle thinking) l yu t cho s tht bi. Do , trc khi dn thn vo nghin cu, cc bn nn t vn quyt nh. Sau y l 15 cu hi m cc bn nn t tr li v quyt nh:
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1. Lnh vc nghin cu ny c quan trng tiu th gi? 2. tng nghin cu c th m rng v ng gp vo s nghip? 3. B mn ti ang lm vic c ph hp vi tng ca ti? 4. tng c phn nh suy ngh ca ng nghip hin nay? 5. Ti am hiu v y vn trong lnh vc ny, v lnh vc no cn khai thc hay tm hiu thm? 6. Nghin cu ca ti c th lp vo khong trng tri thc? 7. c nhiu nghin cu v ti cha, v ti c ng gp g thm? 8. Thi im thch hp cho nghin cu? 9. Nghin cu ca ti c gy tc ng trong chuyn ngnh? 10. Trnh chuyn mn ca ti c ph hp vi mc tiu? 11. Ti c k nng cn thit cho nghin cu? 12. Ti c th gi theo ui d n? 13. Ti quyt tm vo d n? 14. Ti c phng tin trong tay thc hin? 15. Ti c ng nghip c chuyn mn hp tc? Tm li, vit cng nghin cu l mt k nng rt quan trng ca mt nh khoa hc. Xin nhn mnh l rt quan trng (ch khng phi quan trng). cc nc phng Ty, khi mt nh khoa hc c kh nng vit mt cng nghin cu, th l mt chng c v s trng thnh ca nh khoa hc, v l mt nc thang nh khoa hc tr nn c lp. Trc khi kt thc, ti mun k cc bn mt cu chuyn vui nhng hon ton c tht. Hm n, gp anh bn [gi] ng nghip Melbourne, ti hi anh do ny ra sao, anh th di ni Th vn chin u vi cng nghin cu. Anh hi ti, v ti cng ni cng s phn. Anh y ci ni sau khi xong ci bull fighting ny chng ta s hp tc trong mt d n rt ho hng. Anh bn ti xem vic vit vit cng nghin cu c ti tr nh l mt cuc u b; ti v anh y cng ln u trng. C th c hai u tht bi, cng c th c hai u thnh cng, hoc ch mt trong hai thnh cng. Xin ti tr qu tht l mt cuc u tranh. Khng ch u tranh trn "mt trn" tng, m cn mt trn ch ngha! C ch l mt chuyn, nhng dng ch sao cho thuyt phc l mt k nng c ngha sng cn trong cuc u tranh xin ti tr. Xin nhc li rng cng nghin cu khng phi l khoa hc, m l mt cch tip th khoa hc. Tip th bng ch ngha. Ghi nh im quan trng ny vit cng sao cho thuyt phc (ch khng phi khoe) v tng xc sut c ti tr. Chc cc bn may mn! N.V.T Ch thch: 1. y l bi ti vit li t workshop v vit cng nghin cu c thc hin i hc Quc gia TPHCM (VNU) vo nm 2011. Trong workshop th ch c nhng slides, ch cha c bi vit chi tit, v bi ny l mt bi ti liu c thm cho cc bn. Ti mun
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nhn c hi ny by t lng bit n n cc ng nghip VNU, c bit l Trung tm o to v pht trin ngun nhn lc (nay l Vin o to quc t), to iu kin thc hin workshop, m theo cc hc vin l gip ch cho h nhiu. Mt s cn khoe xin c ti tr v hc bng nc ngoi. Xin cm n cc bn hc vin. 2. Cc bn c th copy v phn phi bi ny cho cc bn khc (nu cn). Nhng ti ch mong mun mt iu: ghi ngun. C nhiu khi ti thy mt s khng t bi vit v d liu ca mnh c s dng trong cc hi ngh (v c trn nhng trang bo) m tc gi khng ghi ngun, v iu ny lm ti thy hi bun. Cng sc b ra rt nhiu, thu thp d liu, su tm nhng ca th v, ri rt rut v lao tm son, m ngi ta s dng mt cch v t, khng c n mt ch ghi nhn ch cha ni g n cm n. Ni th thi, ch nu cc bn s dng v c kt qu tch cc l ti vui ri (khng cn cm n u). 3. Ngy 1/10 v 2/10 ny, nhn mt chuyn cng tc H Ni, ti s ni v cch vit cng nghin cu Bnh vin Nng. Ti s c 6 bi ging lin quan n nhng vn bn trong bi ny. Nu cc bn c dp tham d th chc vui lm, v c nhiu c hi tho lun thm. Bi ny tht s c son cho cc hc vin Nng, v son trong mt thi gian rt ngn (ch 4 ting ng h). V qu vi, nn chc cn nhiu thiu st. Nu cc bn pht hin sai st, xin gi th cho ti bit v chnh sa sau ny. Ti hi vng s c dp quay li b sung nhiu chi tit hn, c bit l v d t VN (t cc bn).

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