Cch vit cng nghin cu khoa hc

Nguyn Vn Tun
GS H New South Wales, Sydney
i vi nhng ngi nh ti, vit cong nghin cu gn nh l mt . ngh. Nm no cng
phi vit t nht 3 cong, c khi l on xin bat, cng c khi l on xin ti b nhim. Vit rt
nhiu v tht bai cng rt nhiu. Tht bai nhiu n ni kh m ht! Tht bai gn nh l mt .
qui lut! Nhng cng c thnh cng, d s ln thnh cng t hon s ln tht bai. Chnh qua
nhng tht bai, ti mi hoc oc nhng bi hoc au lng, v l l do tai sao ti mun chia s
cng cc ban tr hon, hay cc ban cha c kinh nghim (hay c t kinh nghim) v cch vit
cong nghin cu. C nhin, ti khng dm ha nu cc ban tun theo nhng g ti hng dn
l s thnh cng, nhng ti dm ha l xc sut thnh cng s cao hon 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 hoc. C nhng vn
khng hn l phc tap, nhng c khi lai rt on gin. Thng, nhng vn on gin l nhng
vn kh nht, v c th dn n nhng khm ph quan trong. Tai sao nam gii hay cht sm
hon n gii? Tai sao ngi dn vng nng thn thng c ln da sm hon ngi dn thnh thj?
Tai sao phu n Vit Nam thch c ln da trng trong khi phu n u chu thch ln da bnh t? Tai
sao cc nam phu thut vin hay chi th? Tn s chi th ca phu thut vin c khc nhau
gia cc b mn? Tai sao bnh nhn t vong nhiu trong hai ngy cui tun? Cht long cuc
sng ca nhng bnh nhn sau khi xa trj ra sao?
l nhng vn tuy on gin nhng i hi chng ta phi suy ngh, tm cu tr li, v trong
vi trng hop, tm gii php. Nhng cu tr li c khi hnh thnh t cm nhn c nhn hoc
long nng bnh dn. Mt nh phu thut, qua kinh nghim lu nm, c th tu tin rng phong
php iu trj 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 thuc t sng n 3 nm sau khi dng mt loai tho
doc, th ngi ta s ngh ngay n tho doc l c ch cho iu trj ung th. Nhng kinh
nghim c nhn, nhng pht hin tnh 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 hoc, bi v cha oc h thng ho.
Mt cch h thng ho vn l qua nghin cu khoa hoc.
1. Suy ngh nh nh khoa hc
Do , ng trc mt vn , mt hin tong, chng ta phi tp cch suy ngh nh mt nh
khoa hoc. Nh khoa hoc suy ngh c phn khc vi ngi thng, v ho t khi no chju su chi
phi ca cm tnh. Mt ca bnh cha thuyt phuc ho v mt liu php iu trj. C trng hop
nhiu ca bnh cng cha thuyt phuc, bi v ho cn phi so snh vi nhm khng oc
iu trj (trong khoa hoc, thng oc goi l "nhm chng control group). Ngoi ra, nh khoa
hoc cn phi pht biu gi thuyt (dua trn cu hi) v kim jnh gi thuyt qua th nghim.
Trong trng hop ung th v doc tho, trc khi i n kt lun nh khoa hoc phi tm hiu co
ch sinh hoc ca doc tho, t gi thuyt v hiu qu, v tin hnh nghin cu thu thp d
liu xem c ph hop vi gi thuyt hay khng. Ni mt cch ngn gon, nh khoa hoc 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 jnh v mt vn . V
mang tnh bt jnh, nn nh khoa hoc phi tm hiu nhng yu t no dn n su bt jnh. 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 tam goi l FNER.
F l vit tt ca feasibility, tc tnh kh thi. Mt cu hi nghin cu tt phi kh thi, tc
nh khoa hoc c kinh nghim chuyn mn, c th tuyn dung y bnh nhn hay i
tong nghin cu, c phong tin o lng v th nghim, v.v.
I l interesting, tc th v. Mt cu hi nghin cu tt phi th vj i vi nh khoa hoc,
xng ng theo ui. C nhiu nh khoa hoc b ra c i ngi chi 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,
phong php mi, hay tng mi. Mt nghin cu chi lp lai y chang nhng g ngi
khc lm th khng c ci g mi, v oc xp vo nhm "me too.
E l ethics, tc o c. Mt nghin cu y khoa phi tn trong quyn con ngi, khng
lm hai ngi, phi bo mt tuyt i (khng oc 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 ao c / y c th d cu hi c th vj 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 oc cu tr li v c th lm thay i chuyn ngnh l
mt cu hi quan trong. "Tc ng y phi hiu l c tc ng tch cuc n thuc
hnh lm sng, n chnh sch y t ca Nh nc, hay c ng gp mt jnh hng
mi. Chng han nh cu hi "Cc chong trnh truy tm ung th v c hiu qu gim
nguy co t vong? l mt cu hi tt v cu tr li s lm thay i nhn thc ca phu 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 hoc 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 thuc t. Mt cu hi nghin cu tt s dn
n gi thuyt khoa hoc hay. Gi thuyt khoa hoc l mt pht biu mang tnh tin l!ng gia
hai hay nhiu bin. Hai ch tin long y rt quan trong! Mt pht biu nh "C mi lin h
gia ung th v v long xong khng phi l gi thuyt v khng c tnh tin long. Nhng
mt pht biu nh "Bnh nhn ung th v c nguy co long xong thp hon bnh khng bj ung
th v th oc xem l mt gi thuyt khoa hoc (v c tnh tin long).
Cng nn phn bit gi thuyt khoa hoc vi gi thuyt thng k. Gi thuyt thng k chi l mt
cch pht biu gi thuyt khoa hoc tin cho vic kim jnh thng k. Nu gi thuyt khoa hoc
l ""nh nhn ti#u $ng c% t& t'ng m( cao h)n ng$i kh*ng " ti#u $ng, th gi thuyt thng
k l ""nh nhn ti#u $ng c% t& t'ng m( "+ng ng$i kh*ng " ti#u $ng. Gi thuyt thng k
va cp cn c khi goi l gi thuyt v* hiu (null hypothesis). Bn canh gi thuyt v hiu,
nh khoa hoc cn pht biu mt gi thuyt o (alternative hypothesis): "nh nhn ti#u $ng c%
t& t'ng m( khc vi ng$i kh*ng " ti#u $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 phong php thng k
oc pht trin ly gi thuyt v hiu lm chun). Mt trong nhng nhim vu 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 hoc khoa hoc Karl Popper, chng ta khng th no chng
minh oc mt gi thuyt khoa hoc.
Sau khi c gi thuyt, bc k tip l lm th nghim (experiment). Th nghim y nn
hiu mt cch rng hon, ch khng on 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 oc chia thnh hai nhm nh (nhm oc iu trj v nhm chng khng
oc iu trj) oc xem l mt th nghim th nghim lm sng. Mt nghin cu bnh chng
cng l mt th nghim. Th nghim l mt bc cuc k quan trong 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 hoc pht biu gi thuyt, ri tin hnh th nghim kim jnh gi thuyt . Mt cng trnh
nghin cu tt thng m ra mt jnh hng mi, v nhiu cu hi nghin cu mi. Chng han
nh cu hi "Tai sao ngi nng thn c ln da sm hon ngi thnh thj, sau khi c cu tr li
(v du) l do vitamin D, th n s m ra hng loat nghin cu mi: tai sao ngi thnh thj hay
thiu vitamin D; yu t no quyt jnh 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 hoach. Nhng qui trnh v k hoach ny phi oc hoach
jnh (trc khi tin hnh nghin cu) trong mt ti liu m ting Anh goi l -./.a'ch 0'o1o/al,
v djch sang ting Vit l " cong nghin cu. Ti mun hiu hai ch cong nh sau:
" l nghj, xut; v "cong l cong lnh. cong nghin cu, do , l mt ti liu
m trong nh khoa hoc nghj cong lnh hay chong trnh lm vic. y l ti liu quan
trong nht trong mt cng trnh nghin cu, v qua m co quan ti tro c th xt duyt cp
kinh ph.
Ti ngh c th xem cong nghin cu nh mt bn v ca kin trc s. Tht vy, c th xem
nh khoa hoc nh l mt kin trc s. Kin trc s phc hoa chi tit xy dung mt cng trnh
trong bn v. Nh khoa hoc phc hoa 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
cong nghin cu nh l mt tc 1h2m ch3 ngh4a khoa hc nhm i tm cu tr li cho cu hi
nghin cu.
l tc phm ch ngha, th cch vit cong nghin cu ng vai tr rt quan trong. Quan
trong l v ngi oc (c th l nh ti tro, hay hi ng xt duyt) dua vo m quyt jnh
cung cp ti tro hay khng. Do , ngi vit (tc nh khoa hoc) cn phi suy ngh rt cn thn,
c h thng, v s dung ngn ng cho chnh xc. Ni th c l d, nhng thuc hnh th khng d
cht no. Ti liu ny c muc ch khim tn l chia s cng cc ban nhng kinh nghim v cch
vit cong nghin cu y khoa sao cho thuyt phuc v nng cao xc sut oc ti tro.
Vit cong nghin cu khc vi vit bi bo khoa hoc. Trc y, ti vit mt loat bi v
cch vit mt bi bo khoa hoc (c th xem trang nguyenvantuan.net trong phn " K nng
khc). Nhng phong php vit bi bo khoa hoc khng th p dung cho vit cong nghin
cu. Nhng khc bit chnh l muc tiu v thi gian tnh. Muc tiu ca vit cong nghin cu
l thuyt 1h5c nh ti tro hay hi ng xt duyt rng chng ta c tng tt, c cch tip cn
hay, c phong tin thuc hin nghin cu. Muc tiu ca vit bi bo khoa hoc l bo co
nhng pht hin rt cu th trong mt nghin cu v nhng pht hin ny c ngha g. V thi
gian tnh, vit cong l phn nh v tong lai, cn vit bi bo khoa hoc l bo co nhng g
lm trong qu kh. Vit cong mang tnh ha h6n, nhng ha hen mt cch khoa hoc
(tc c bng chng), do rt khc vi vit bo co khoa hoc thng mang tnh bin minh v
bin lun.
Theo kinh nghim c nhn, ti thy vit cong nghin cu cung cp cho mnh nhiu co hi
rt hay. Th nht l co hi sp xp tng mnh mt cch c h thng, c trc, c sau.
Nhng tng hn n, khi oc m t trn trang giy s lm cho chng ta suy ngh logic hon.
Th hai, vit cong cng l mt co 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 cong nghin cu
cng c ngha l tm ng nghip mi. Nhn nhn nh th, vit cong c nhiu loi ch, ch
khng phi chi l "v vi theo cch ni mia mai ca ngi ngoi cuc.
Vit cong (hay vit vn ni chung) l mt cch suy ngh. C l chng ta u ng rng vit
vn l mt phong tin chia s thng tin vi nhng ngi quan tm. Ngi 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 1h)ng tin hay mt c*ng c5 suy ngh. Suy ngh cng cn phi c phong tin,
v theo ti vit xung nhng cu ch do mnh lua chon chnh l mt phong tin. C l chnh v
th m Nh vn [v cng l bc s] Verghese tng ni mt cu bt h, "7 8'it. to un9.'/tan9
8hat 7 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 cong nghin cu l "tt hay
"d. Kinh nghim c nhn ti cho thy c 5 tiu chun: t'ong /ng, )n gin, chnh :c, khch
;uan, v c<u t'c logic.
Trong sng c ngha l trnh nhng cu vn rm r, nhng t kh hiu. Nu vit " =>ng ?
in/ulin @ nh%m iAu t' cao h)n th s khng r rng, v ngi oc khng bit cao hon nhm no.
Ngay c cch vit "=>ng ? in/ulin @ nh%m iAu t' cao h)n nh%m chngB cng c th ni l cha
at, bi v cu vn hm ni rng tt c bnh nhn trong nhm iu trj u c nng insulin
cao hon nhm chng mt tnh hung rt kh xy ra. Trong thuc t th "Cnh t'ung "Dnh, n>ng
? in/ulin @ nh%m iAu t' cao h)n nh%m chngB c l r rng hon.
Vn phong khoa hoc rt khc vi tho. Ti thng ni a [v hay ly lm v du] v cu tho ca
mt tc gi ti rt mn m (L at): C*i tha th2n gi3a chEa Fun =g3 G Li chuc tui mnh ni
tht !h"i sinh, Cu trch trong tp tho H$ng Ih3. l mt cu vn rt kh hiu! Mt thi s
khc ti cng rt ngng m l Hong Cm, ngi sng tc tho mt cch siu thot. ng c
sng tc bi CDnh IJm, trong c hai cu rt du dong: =u anh cKn t'L nh nMm cN G Fuyt
%n .m vA /Ong vi anh. C th ni l nhng cu tho c o, gieo vn tuyt vi, m iu rt
nhac tnh, nhng nh khoa hoc khng th vit nh th oc. Nh khoa hoc phi vit vn trong
sng. Chng han nh nu ni cKn t'L th phi jnh long r rng bao nhiu tui l tr; nu
ni nMm cN th phi ni nm no; nu nivA /Ong vi anh th phi ni sng u. Vn phong
khoa hoc khng th chung chung oc. Nh khoa hoc khng phi nh tho. Nhng rt tic, nhiu
khi ti bt gp nhng cu rt tho trong nhng bi bo khoa hoc t ng nghip Vit Nam. Mt
trit gia c tng ph bnh ngi ng rng chng ta [ngi ng] hay ln ln gia tho v
khoa hoc!
Trong vn phong khoa hoc, mt cu vn phi c thng tin. Cu vn khng c thng tin l cu vn
tha. V du tiu biu cho cu vn khng c thng tin l [hay thy trong gii bo ch] "I*ng t'Dnh
nghin cu c% P m5c tiu. v oc xong cu vn ngi oc khng c bt c mt ni dung no c;
ngi oc k vong muc tiu l g v tu hi tai sao tc gi khng vit ra. Nhng nu vit
"I*ng t'Dnh nghin cu c% P m5c tiuQ :c nh nh h@ng cRa can thi1, vS :c nh yu tO
nguy c) th l mt cu vn c thng tin. Th oc cu:
=goSi "a kha cnh t'n, chng t*i cKn 1hn tch v<n A 9Ta t'n l thuyt vMn ho :U h?i,
V y, li c% nhiAu kha cnh khc,
Hai cu vn ny c th thch hop cho bo ch, nhng hon ton tht bai trong khoa hoc! Ni cch
khc, trong vn cnh khoa hoc, mt cu vn phi tn tai mt cch ring l (self-contained), c
thng tin v c ; oc cu vn nh th ngi oc khng hn cn phi oc 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 oc k c th vit ngn gon hon. Thay v dng
nhng danh t gc Hn, chng ta nn c gng dng nhng danh t gc Vit.
Chnh %c l jnh long ho ni dung thng tin. Trnh nhng t ng m m. Ting Vit ta (v
ting Anh cng th) c nhng ch nh khong, :<1 :&, ?, gJn, a /O, 1hJn ln, n%i chung,
v,v, khng mang tnh jnh long cao. Khoa hoc l cn o ong m, nn c gng vit mt cch
jnh long. Kh bit bao nhiu l a /O, hay l a s? Thay v vit "a s bnh nhn
., chng ta nn vit (nu c s liu) " bnh nhn . th s r rng hon. Trong khoa hoc
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 "Cha anh,
n%i chung lS .., ng Thanh ngt li ngay: =%i 'ing, ch kh*ng n%i chung, Hi trng ci xo.
ng Nguyn B Thanh p dung tiu chun khoa hoc vy.
&hch 'u"n l cch vit phi cm tnh, v nht l khng "nht ch vo ming ngi oc. Thay v
vit "Su khc bit'<t c ngha lm sng, th nn vit "Su khc bit c ngha lm sng ri
trch dn con s hay d liu ngi oc phn xt. Khch quan cng c ngha l trnh gi jnh
(kiu nh Wi cNng "it '+ng .). Tun theo nguyn tc khch quan cn c ngha l trnh nhng
cu vn khng c chng c.
C(u tr)c *ogic 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 5 nhng hiu lm l do cu trc oan vn, chi c
5 hiu lm l do ni dung. Do , c l y l tiu chun quan trong nht trong cch vit
cong. Bt c khi nim g mi cn phi c gii thch trc . Nu mt oan vn xut hin cum
t "cht long cuc sng m khng oc cp n trong cc oan vn trc l mt cch vit
rt . d (thiu tnh khc chit).
Cu trc cu vn v oan vn nhn manh mt ch . Chng ta th oc cu vn sau y:
a, XYc 9E thuOc c% hiu ;u cao, nhng cNng gy tc hi ng k#,
", XYc 9E thuOc gy tc hi ng k#, nhng c% hiu ;u cao,
c, ChuOc c% tc hi ng k#, nhng '<t hiu ;u,
d. ChuOc c% hiu ;u cao, nhng gy tc hi ng k#
Ci khc bit chnh gia cc cu vn c l khng phi ni dung, m l vj tr nhn manh tng.
Cu hi t ra l trong mt cu vn, cn nhn manh phn no: phn u, phn gia, hay phn
cui? Cc chuyn gia v vit vn khuyn co rng nn dng phn u nhn manh im chnh
yu. (Cng c chuyn gia nghj nn dng phn cui cu vn nhn manh). Nn m u cu
vn vi mt t kho, nu chng ta mun ngi oc ch n ch . Trong cu loUng :)ng
lS m?t v<n A y t ;uan t'ng, ti nhn manh n long xong. Tuy nhin, nu ti vitX?t t'ong
nh3ng v<n A y t ;uan t'ng nh<t hin nay lS loUng :)ng, th cng ni ln ci , nhng c l
ngi oc s hiu rng ti ang nhn manh n kha canh y t hon l long xong.
Vj tr nhn manh v cch dng c khi lm cho ngi oc hiu khc. Trong mt cun sch v
cch soan cu hi trong nghin cu khoa hoc, tc gi k chuyn [vui] v thy tu, mt ngi tu
dng a Minh (Dominican) v mt ngi tu dng Tn (esuit), 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, ho gp lai, v c cuc i thoai nh sau:
Vj tu s dng a Minh hi vj tu s dng Tn: Ch "A t'n cRa anh n%i gDZ
Vj tu s dng Tn: =gSi n%i ok,
Tu s dng a Minh: [ui nh&\ ]A t'n cRa t*i n%i '+ng % lS m?t t?i,
Tu s dng Tn: Ch anh hi "A t'n cRa anh nh th nSoZ
Tu s dng a Minh: C*i hi ngSi '+ng h)t thu+c * trong !hi c,u nguyn c% ok kh*ngZ
Tu s dng Tn ln ri ni: Wh, cKn t*i hi ngSi '+ng c,u nguyn trong !hi h)t thu+c * c% ok
kh*ng,
l mt bi hoc ng ch v cch t ch vo vj tr thch hop. Chi cn thay i th tu ca
ch l c th thay i ngha ca cu vn, v lm cho ngi oc c th hiu khc. Trong vit
vn khoa hoc, tc gi khng nn cho ngi oc hiu hai ngha!
Ngoi ra, nn trnh cch vit "nh trng b di. l cch vit tuyn b mt ch , nhng
nhng cu sau lai ni v ch khc. Th oc oan vn:
^Uy c_ :)ng Ei lS h`u ;u hay gY1 ng hSng th a cRa loUng :)ng, "nh c% t& l tb vong vS
tSn 1h cao, ch<t l!ng /Ong cRa ng$i "nh gim ng k# ngay c khi U !c iAu t' ng
cn, ]nh th$ng gY1 @ ng$i cao tu_i vi t& l n3Qnam lS aQd,
Cu u ca oan vn trch dn trn c 3 chnh: su ph bin, h qu, v iu trj. oc xong cu
vn , ngi oc k vong 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, oan vn trn cha at. l cha ni n cch dng ch
v ngha cn kh m m (v du: "ng hng th 3 ca long xong c ngha l g). Trong vn
phong khoa hoc, mi cu vn chi ni ln m thi; tham vong qu s khng chuyn ti ht
ngha.
4. Ni dung mt cng nghin cu
Mt cong nghin cu bao gm nhng g? Tr li cu hi ny cn tu thuc vo "vn ho v
qui jnh ja phong. Vit Nam, nhng cong nghin cu thng c nhng muc sau
y:
Muc tiu ca ti;
Tng quan tnh hnh nghin cu v tnh cp thit ca ti ;
Ni dung nghin cu;
Phong php nghin cu, cch tip cn vn ;
Hop tc quc t (nu c) ;
Tin thuc hin;
Dang kt qu du kin ca ti;
u cu khoa hoc i vi nhng sn phm du kin tao ra;
K hoach trin khai;
Loi ch mang lai v tc ng ca kt qu nghin cu;
Cc t chc tham gia nghin cu; v
Kinh ph.
Nhng muc ca mt cong nghin cu trn y khng c vn g, nhng cch t chc
v sp xp thng tin th hoi khc so vi nc ngoi. Nhng muc 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 (cu th l c, Anh, c, Thi Lan, M, v.v., nhng noi m ti c kinh nghim) th ni dung
mt cong nghin cu thng on gin hon Vit Nam. nhng nc , mt cong
nghin cu bao gm nhng phn sau y:
Tm loc (synopsis): ging nh mt tm tt cho mt du n nghin cu. Thng thng
chi - trang giy;
Muc tiu cu th (specific aims);
Bi cnh v tm quan trong (background significance);
Kt qu nghin cu so khi (preliminary results);
K hoach nghin cu v phong php (research design and methods); v
Ti liu tham kho.
Tuy nhng muc ca cong Vit Nam c v hoi nhiu, nhng s trang thng chi dao
ng trong khong n trang. nc ngoi, nh NH (Vin t ca M chuyn cp ti tro
cho nghin cu y sinh hoc v cng l mt trung tm nghin cu) th c qui jnh s trang rt r
rng. Mt cong ca NH ti a l 3 trang (hin nay th c thay i v gim xung cn
trang?) Ti c lm mt so snh gia mt cong tiu biu ca VN v ca NH th thy nh
sau:
So snh s trang ca mt cng VN v NIH
mc Vit Nam M (NIH)
Tng quan ti liu
Muc tiu
Kt qu so khi -
Phong php 5 -
Ni cch khc, cong nghin cu ca VN vit di v bi cnh v tng quan ti liu, nhng rt
ngn v phong php. Ngoc lai, nc ngoi (tiu biu l NH ca M), mt cong nghin
cu ch yu l phn phong php (rt di) nhng phn tng quan ti liu th ngn. Rt nhiu
cong nghin cu t VN m ti oc qua, trong phn tng quan ti liu, tc gi vit di nhng
chng lin quan g n ch v muc tiu nghin cu! Hu nh cong nghin cu y khoa
no cng vit theo mt cng thc, trong c c phc iu trj, nhng jnh ngha rt cn bn
v bnh, nhng yu t nguy co (m c l ai trong ngnh cng bit). C khi cong nghin cu
tm hiu ti l mc bnh, nhng tc gi phi im qua mt cch kh di dng v phong php
iu trj! Phn ln nhng thng tin ny tht ra l tc gi djch t sch gio khoa, hoc djch 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 cong nghin cu nc ngoi phi c phn "Kt qu so
khi, cn Vit Nam th khng c hoc khng yu cu. c v M, mt cong m khng c
kt qu so khi (hay nhng nghin cu trc y cng ch m nh khoa hoc lm) th
khng bao gi oc qua vng u xt duyt, rt rt kh c kh nng oc ti tro. Trong cc
phn sau y, ti s trnh by cch vit phn quan trong nht: muc tiu, bi cnh, nghin cu
so khi, v phong php nghin cu. Ti s c gng a vi v du cu th t nhng cong
trc y ca ti v ca cc ng nghip khc. V bi ny cng nhm n cc ban nghin cu
sinh ang hay sp i hoc nc ngoi, nn ti cng trnh by vi v du bng ting Anh cc ban
c th tham kho.
4.1 Phn mc tiu nghin cu
Muc tiu nghin cu l phn quan trong ca mt cong nghin cu, v l "b mt m
ngi oc s nhn qua. Khi ngi oc thy muc tiu nghin cu c ci g mi hay th vj th ho s
oc tip; nu khng, ho c th xp lai cong v th l tc gi tht bai. Do , c th xem
phn muc tiu nghin cu nh l mt ci test cho ngi oc. Chi khi no ci test ny oc qua
th tc gi mi thnh cng mt phn trong vic thuyt phuc ngi oc.
Phn muc tiu nghin cu nn oc cu trc 3 phn nh sau:
Mt oan vn tng qut v vn chung;
Pht biu v muc tiu chung (ting Anh goi l goal);
Pht biu v muc tiu chuyn bit (specific aims), v mi muc tiu nn km theo mt gi
thuyt.
oan vn tng qut phi ni oc v<n A ln m nh nghin cu quan tm. l mt oan vn
mang tnh holi/tic,hay bi cnh nghin cu m cong ny s ng gp mt phn tri thc.
Muc tiu chung hay muc tiu tng qut l m5c tiu lu 9Si m nh nghin cu mun at oc.
C th nghin cu hin nay cha at oc, nhng s ng gp mt phn trong jnh hng gii
quyt vn v lu v di. Trong bng , muc tiu ti hu v tng qut l ph li; tong tu,
trong nghin cu khoa hoc, muc tiu tng qut l nhm n gii quyt mt vn chung cho
chuyn ngnh.
Muc tiu chuyn bit l nhng tm im cu th. Cu th hiu theo ngha c th o lng oc, c
th kim jnh oc. Ngi vit cong kinh nghim thng vit mi muc tiu chuyn bit km
theo mt gi thuyt. Thinh thong, nu c "t, nh nghin cu nn km theo nhng kt qu du
kin cho tng muc tiu. Thng thng chi cn -3 muc tiu chuyn bit l . Nhiu muc tiu
qu s kh thuc hin; t qu s khng p ng tiu chun ca mt du n nghin cu khoa hoc.
Trong thuc t, ti tng thy nhiu cong tht bai (k c ca chnh ti) v cch pht biu
muc tiu cha at. Nhng sai lm ph bin trong cch vit muc tiu chuyn bit bao gm:
Muc tiu phi thuc t, tc thiu tnh kh thi;
Muc tiu khng c l do chnh ng (ging nh t "trn tri roi xung);
Muc tiu mang tnh qu m t (nh tm hiu bao nhiu nam v n c hi chng chuyn
ho);
Muc tiu qu phc tap, m oc ln ngi ta khng hiu tc gi mun lm g;
Muc tiu qu phu thuc ln nhau (s bn di y).

V d 1: y l trch dn cong nghin cu v long xong ca mt ng nghip v c su
ng ca tc gi.
[Hon vMn t_ng ;ut # e9Du 9ctB ?c gi '+ng y lS v<n A ;uan t'ng]: Long xong l mt
bnh m luc ca xong bj suy gim, cht long xong xung cp, v h qu l tng nguy co
gy xong. H qu ca long xong l gy xong. Chn on long xong dua vo o lng
mt xong. Tuy nhin, Vit Nam cha c gi trj tham chiu cho vic chn on long
xong.
[X5c tiu t_ng ;ut 1 ng v<n A chung]: Muc tiu lu di ca nghin cu ny l pht trin tiu
chun chn on long xong cho ngi Vit (nam v n) v xy dung m hnh tin long gy
xong. Hon thnh muc tiu ny s gip cho vic chn on long xong chnh xc hon, v
nhn dang nhng c nhn c nguy co gy xong cao can thip sm v gim qui m long
xong trong cng ng.
[X5c tiu chuyn "it] at oc muc tiu , chng ti nghj theo ui cc muc tiu cu th
nh sau:
Mc tiu 1: y dung gi trj tham chiu mt xong i v xong ct sng nam v
n; v qua , xc jnh chi s C cho chn on long xong ngi Vit.
Gi thuyt 1: Mt xong ngi Vit thp hon mt xong ngi u M; do ,
chi s T ca ngi Vit cng khc vi chi s T ca T chc t Th gii.
Kt qu k vng: Gi trj tham chiu v mt xong (trung bnh, lch chun, chi s
T) c th s dung cho vic chn on long xong nam v n Vit Nam.
Mc tiu 2: c jnh ti l lu hnh long xong v gy xong nam v n;
Gi thuyt 2: N gii c nguy co long xong v gy xong cao hon nam gii, nhng
khc bit s gim dn trong tui trn .
Kt qu k vng: Kt qu l tn s long xong v gy xong theo gii v tui.
Nhng kt qu ny s cung cp chng c v qui m long xong v gy xong ngi
cao tui v sau mn kinh (n).
Mc tiu 3: c jnh yu t nguy co lin quan n long xong v gy xong, k c
cc markers chu chuyn xong, v qua pht trin m hnh tin long long xong v
gy xong.
Gi thuyt 3: Marker chu chuyn xong tng theo tui, v chu chuyn xong cao
tong quan vi su suy gim mt xong nam v n.
Kt qu k vng: Gi trj tham chiu v marker chu chuyn xong, v m hnh tin long
gy xong cho ngi Vit. Vi m hnh ny, cc bc s c th c tnh nguy co gy
xong ngay c trong iu kin khng c my DA.
V d 2: Trong v du ny, tc gi vit phn muc tiu rt "bi bn.
[X@ Ju lS v<n A 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. et, 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 inury. Accordingly, infusion of apoE has
been shown to decrease LPS-induced morbidity and mortality in rodents. [, 3] Also, apoE-
deficient mice have an increased susceptibility to lethal infection when inected with live bacteria.
[, 5] But, unexpectedly, our laboratory has recently discovered that infusion of apoE inc'.a/.9
rather than decreased mortality after cecal ligation and puncture, an in vivo model of
polymicrobial sepsis. [] 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. [9Eng gch 9Kng # nh<n mnh m5c
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.
[0ht "i#u gi thuyt chung] ur 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. [-] Third, triglyceride-rich lipoproteins clear LPS from the circulation and
deliver it to the liver [, ], where lipoprotein-LPS complexes subsequently modulate the hepatic
immune response to infection. [-] 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.
[0ht "i#u gi thuyt chuyn "it] The specific hypothesis driving the proposed research is
that apoE, a key constituent of trigIyceride- rich Iipoproteins, reguIates the host response
to severe infection through its effects on NKT ceII 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 Iong term
goaI 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.
[X5c tiu chuyn "it, 9Eng t* `m # nh<n mnh vS gy ch f, C'ong mgi m5c tiu, tc gi ch&
'a h /h c% kt ;u gD] The specific aims of the proposal are therefore:
1. To demonstrate that serum apoIipoprotein E (apoE) concentrations correIate with
morbidity and mortaIity in a murine modeI of poIymicrobiaI 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 Th cytokine secretion, liver
inury and bacterial clearance;
2. To demonstrate that apoE promotes the activation of naturaI kiIIer T (NKT) ceIIs 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
mortaIity 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 cong nghin cu lm sng M. Tc gi c cch vit c ong hon. M
u, tc gi m t mt cch ngn gon v ni dung ca nghin cu. Sau , tc gi pht biu muc
tiu chnh (primary aim) v muc tiu phu (secondary aim).
[X@ Ju, m* t ngcn gn vA c*ng t'Dnh 7-7i vS gi thuyt] The nsulin Resistance ntervention
after Stroke Trial (RS) is a randomized, double-blind, placebo-controlled trial that will test the
hypothesis that reducing insuIin resistance and its sequeIae with thiazoIidinedione
therapy wiII prevent stroke and myocardiaI infarctionamong patients with a recent ischemic
stroke. Eligible subects are men and women over years of age without diabetes mellitis who
have insulin resistance and a non- disabling ischemic stroke. During 3 years of recruitment, 33
patients will be randomly assigned to pioglitazone, a thiazolidinectone, or placebo.
[X5c tiu chuyn "it] 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 M among non-diabetic men and women over age 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 M) within four years from to
. The basis of this hypothesis is research showing... j9.tail/ a"out /tu9i./ a//ociating in/ulin
'./i/tanc. 8ith inc'.a/.9 'i/k ko' /t'ok., X7, .tc,l,,,]ythese and other mechanisms, we
hypothesize that pioglitazone will protect patients with ischemic stroke and insulin resistance
against recurrent vascular events.
Trong cc v du trn y, d dng thy rng cc muc tiu chuyn bit tuy nhm n mt muc
tiu tng qut, nhng khng l thuc vo nhau. y l mt chin loc trong vic soan cong
nghin cu. Nu muc tiu tu thuc vo su thnh bai ca muc tiu , v muc tiu 3 phu thuc
vo kt qu ca muc tiu , th n s rt . nguy him. Nguy him l v nu muc tiu trc
tht bai, th tt c cc muc tiu khc u khng th thnh hin thuc, v do , cng trnh nghin
cu s khng kh thi. Bi hoc y l cn phi thit k nghin cu sau cho cc muc tiu chuyn
bit t phu thuc vo nhau.
4.2 Phn bi cnh v tm quan trng
Phn bi cnh v tm quan trong, nh tn goi, c chc nng "dn dung sn khu cho cng trnh
nghin cu. Khng chi dn dung, m cn phi ni ln oc tm quan trong ca cng trnh
nghin cu. Khng ai mun u t vo nhng cng trnh nghin cu khng gy nh hng hay
tc ng tch cuc. C phn chnh l bi cnh (background) v tm quan trong (significance):
Trong phn bi cnh, tc gi cn phi vit theo phong cch k chuyn. Nhng k chuyn mt
cch khoa hoc, sao cho ng nghip khng cng chuyn ngnh vn c th hiu oc vn .
Mi mt muc tiu chuyn bit trong phn u cn phi oc l gii trong phn bi cnh. Phi vit
v thuyt phuc ngi oc s oc tip, v mun at muc 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 chi ra cho oc
khong trng tri thc (cha bit), t thnh cu hi cho nghin cu. V du chng ta bit
tnh trang thiu vitamin D cc tinh 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 ti l thiu hut vitamin D c dn
pha Bc l bao nhiu.
V cch vit, khng chi on gin lit k nhng nghin cu trc, m phi dng "phong php
C. Phong php ny vit tt t ng t:
Com-"r. so snh: tc gi cn phi so snh nhng thng tin t nhng nghin cu
trc;
Contr"st i chiu: sau i chiu v gii thch tai sao c su khc bit;
Cit. trch dn: iu bt buc l nu dng d liu ca ng nghip th phi trch dn; v
Criti'u. ph bnh: "Ph bnh y c ngha l ph bnh mt cch knh trong, 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.

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 thuc hnh lm sng;
Phong php mi; v
Tri thc s rt ra oc t cng trnh nghin cu.
Phn tm quan trong c khi phi dng n k thut "pitch (ln giong). Ni cch khc, cn vit sao
cho ngi khc c th trch dn mt cu t cong. Chng han nh nu l nghin cu v di
truyn, ti c th vit
"Ch. /tu9i./ in thi/ 1'o1o/al 8ill 1'ovi9. a "a/i/ ko' un9.'/tan9ing all.lic h.t.'og.n.ity
inklu.ncing clinical .n91oint/, ultimat.ly im1acting on 9i/.a/. 9.v.lo1m.ntB jnh3ng nghin cu
m* t t'ong A c)ng nSy /h cung c<1 nAn tng # hi#u "it vA /T a 9ng al.n c% nh h@ng
n kt c5c lm /Sng, vS /au cEng lS tc ?ng n /T tin t'i#n cRa "nhl,
Mt cu nh th i vi vi ng nghip Vit Nam s goi l "n, nhng nc ngoi th hon
ton c th chp nhn oc. Vn cn tu thuc vo vj tr ca tc gi v uy tn trong chuyn
ngnh. Nghin cu sinh c l khng nn vit khim tn hon, nhng vi ngi c "tn tui th
mt cu pitch nh th hon ton bnh thng. Nn nh rng /it 01 c2$ng *3 mt cch 4n 5
t26ng, nn tc gi cn phi thuyt phuc tm quan trong ca nghin cu.
V d 4 : Trong cong cu di y, tc gi mun thuyt phuc ngi oc v tm quan trong
ca cng trnh nghin cu:
Long xong v gy xong l mt vn y t cng ng ln nc ta, v hng nm c khong
. ngi gy xong, dn n gim tui tho v han ch lao ng. Mt xong l mt chi
s lm sng quan trong v M c th tin long nguy co gy xong cho mt c nhn. V th,
M cn oc s dung chn on long xong.
Cng trnh nghin cu ny c muc tiu xy dung gi trj tham chiu M cho phu n v n ng
Vit Nam. Vi gi trj tham chiu ny, vic chn on long xong ngi Vit s chnh xc hon
v qua chng ta c th bit oc qui m long xong nc ta.
Do , cng trnh nghin cu mang tnh cp thit, v su hin din ca my DA nhiu nc ta
nhng cha c gi trj tham chiu cho ngi Vit. V th, kt qu nghin cu c gi trj thuc tin,
c th p dung ngay cho vic chn on long xong. ngha l lun ca cng trnh nghin cu
l cung cp nhng thng tin khoa hoc cho vic hoach jnh cc chin loc phng chng bnh
long xong qui m cng ng.
V d 5 : Trong cong di y, tc gi vit ngn gon v bi cnh v tm quan trong nghin
cu, bng cch dng cc tiu nh nhn manh tng im:
7hy 8tu9y 8'u"r. C.** :is."s. in th. &i9n.y;
Renal dysfunction commonly complicates square cell disease and is a maor 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 (5), while chronic uremia, filtration insufficiency, and renal
vascular disease occur in - of adults with square cell disease (, 5). 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-occIusion and infIammation in the kidney
are IikeIy to be different from those in other organs, there is a criticaI need for basic
research on square ceII disease that focuses on the kidney.
<hysio*ogic"* :.t.rmin"nts o= >"so?occ*usion in th. &i9n.y
Research on renal vaso-occlusion is limited to two studies that suggest severe medullary
hypertonia causes sequestration of SQ RBCs in the kidney (3, ). 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. nSpecific Aim
1, we adapt the isolated rat kidney model used in these original studies (3, ) to determine
effects of tubular hypertonia, and mixed arteriolar hypertonia and renal inflammation on kidney
micro vaso-occlusion...etc.
8umm"ry "n9 C*inic"* 8igni=ic"nc.
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
cong nghin cu Vit Nam thng khng c phn ny! Nhng i vi cc co quan ti tro
nghin cu khoa hoc nc ngoi th y l phn khng th thiu oc. Tuy ni l "nghin cu
so khi, nhng trong thuc 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 co s cho cu hi nghin cu v pht biu
gi thuyt.
Phn kt qu so khi cn c muc ch quan trong khc l thuyt phuc ngi oc rng tc gi c
kinh nghim. Qua phn nghin cu so khi, ngi oc c th nh gi tc gi hay nhm nghin
cu c sn k thut, phong php, hay cng ngh cn thit thuc hin cng trnh nghin
cu. C l quan trong hon l co quan ti tro cm thy thuyt phuc rng tc gi c th thuc hin
nghin cu (ho an tm "chon mt gi vng!) Thng thng, phn ny chim khong -
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 so khi l trnh by theo tng muc tiu
chuyn bit. Trong v du di y, tc gi trnh by
PreIiminary Data
This proposal is a collaboration between the P and Dr. Barry Hurlburt in the Department of
Biochemistry and Molecular Biology. The collaboration takes advantage of the expertise of the P
in the molecular genetics of S. aureus and the biochemical expertise of Dr. Hurlburt in
transcription factor structure and function (,5). The overall goals are ) correlation of the
expression of the sarA, sarB and sarC transcripts with the production and activity of SarA, )
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 obectives. 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 ) and define the sequence characteristics required for the regulation of cna
transcription (Specific Aim ), (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.
PreIiminary 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 RN3. tilizing Nde
and BamH restriction sites incorporated into the oligonucleotide primers, the fragment containing
the sarA coding region was cloned into the E. coli expression vector pETA. Because the Nde
site (CATATG) in the vector overlaps an ATG start codon, cloning of the sarA coding region into
the Nde 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
pETA 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 BL(DE3)pLysS.
Transformants were grown to mid-log phase before inducing SarA expression by adding PTG to
a final concentration of . 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. ).
PreIiminary data for Aim #2. Purification of SarA. A 5 ml culture of the BL(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 subected to a
series of ammonium sulfate precipitations culminating at saturation. The pellet from each
precipitation was resuspended in SDS-PAGE buffer and examined along with an aliquot of the
supernatant (Fig. , left). The supernatant remaining after the final precipitation was found to
contain SarA.
4.4 Phn phng php (reseach approach)
Sau phn X5c tiu, ]Oi cnh m tJm ;uan t'ng, =ghin cu /) kh@i, l phn 0h)ng 1h1, y
l phn di nht v chi tit nht so vi phn trc. Muc ch ca phn 0h)ng 1h1 l thuyt
phuc ngi oc rng nh nghin cu:
C k hoach tt kim jnh gi thuyt t ra trong phn X5c tiun
C kin thc, k nng, v phong tin thuc hin cng trnh nghin cu;
ngh n nhng tnh hung xu s gp phi v c k hoach i ph; v
Din gii kt qu du kin mt cch khch quan.
Nn nh rng trong phn phong php, tc gi phi 9@ !in tnh hu+ng 4(t *Ai s xy ra trong
khi thuc hin nghin cu. Khng mt nghin cu no u oc tin hnh mt cch "thun bm
xui gi c. Bt c nghin cu no cng c vi truc trc, khng ln th nh, v c th nh
hng n vic thuc hin cc muc tiu. Chng han nh trong nghin cu v long xong, c
th nh nghin cu s gp kh khn nu my DA h hng, hoc bnh vin thay i k thut
vin, hoc cc mu sinh phm bj 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 oc 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
hoach i ph.
Cch vit hiu qu nht cho phn phong php l vit cho tng muc tiu. Nu cong c 3
muc tiu chuyn bit, phn phong php phi c 3 phong php tong thch. Cu trc phn
phong php c th l:
D. K hoach th nghim
D.1 K hoach cho mc tiu 1
D.. Thit k, l do, tm quan trong
D.. Phong php cho muc tiu
D... Ci mi
D... Han ch
D...3 Kh khn c th tin on trc
D... K hoach khc phuc
D...5 H qu
D..3 Phn tch d liu
D.. Din gii kt qu tin on trc
D.2 K hoach cho mc tiu 2
[.]
D.3 K hoach cho mc tiu 3
[.]
V d 6 : Trong cong di y, tc gi m t phn phong php c th ni l rt y .
Ngi oc gn nh bit chnh xc tng bc tc gi sp lm g, tai sao lm, lm nh th no, v
kt qu du kin ra sao.
Research Design and Methods:
The first specific aim of this grant application is to use the HFM 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 HFM
system under these conditions have the same mutations as those that emerge when people are
treated with these drugs. n the second specific aim, we will use the HFM 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 HFM 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 (-3). Treatment of patients with influenza with the neuraminidase inhibitors,
oseltamivir carboxylate or zanamivir, usually does not lead to the emergence of resistant viruses
(). 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 (-).
Data presented in the preIiminary resuIts section of this grant application showed that
treatment of MDCK cells infected with a clinical isolate of influenza A virus in the HFM 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 HFM system. The purpose of this portion of the grant application is to confirm these
observations with A/Albany// 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. ExperimentaI Design. We will examine the effect of amantadine and oseltamivir carboxylate
on the replication of wild type rgA/Vietnam/3/xA/PR//3 (a surrogate for avian H5N
influenza virus), A/Texas/3/(HN), A/Sydney/5/(H3N), and A/Victoria/3/5(H3N) in the
HFM system. For comparison, we will also include our original clinical isolate,
A/Albany//(H3N), to be certain that our original observations are reproducible for amantadine
and oseltamivir carboxylate. seltamivir carboxylate will be tested against B/Lee/ and
B/Memphis// viruses. [.]
C. Expected resuIts. Resistance will emerge under monotherapy. Amantadine resistant strains
will have mutations in the M gene (residues , , 3, 3); neuraminidase inhibitor resistant
strains will have mutations in the NA gene (residues and ) and/or HA genes (multiple
residues).
D. PotentiaI probIems. t is often difficult to generate mutations in vitro in the 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 A5
pulmonary alveolar epithelial cells (), St ude porcine lung (SPL) cells (3), STGal cells ()
or SAT cells (5) which express cell surface receptors with more terminal sialic acid, and Mink
lung cells () to perform these dose ranging studies aimed at producing resistant viruses in the
HFM system. t 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. f this grant application is funded we will be able to purchase 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/3/5, A/Texas/3/, rgA/Vietnam/3/xA/PR//3, and A/Albany// and
oseltamivir carboxylate for B/Lee/ and B/Memphis// in the HFM system. Each experiment
will be repeated at least time. ne hollow fiber experiment takes approximately two weeks to
perform from setup to take down. Analysis of virus yield (plaque assay, TCD5 assay and real
time quantitative PCR) will take an additional two weeks. Therefore, each experiment, including a
repeat, will take approximately 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 hollow fiber experiments. Since we can study
two viruses at a time for one drug or one virus for two drugs, Specific Aim will take at least one
year to complete.
cong nghin cu oc vit cho i tong l ng nghip, nhng l ngi ng vai tr bnh
duyt. Cu hi t ra l ngi bnh duyt k vong g khi oc mt cong nghin cu. Bit oc
k vong ca ho cng l bit cch p ng. Ti t mnh vo vai tr ngi duyt cong, v
ti s k vong nhng iu qua nhng cu hi sau y m ti mun tm cu hi:
tng th vj, cch tn, c th ng gp cho chuyn ngnh hay khng?
D liu so khi c "manh hay thuyt phuc 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 tuu ca tc gi ra sao? Trong yu t ny, ti mun xem
qua thnh tch trong thi gian 5 nm gn y;
cong oc soan mt cch r rng, logic, v chi tit hay khng?
Cch vit trong sng v gon (v iu ny phn nh t duy ca tc gi).
Nghin cu khoa hoc i hi suy ngh v tnh ti mi. Khoa hoc khng chp nhn suy ngh hi hot.
Nhng suy ngh m m (muddle thinking) l yu t cho su tht bai. Do , trc khi dn thn
vo nghin cu, cc ban nn tu vn quyt jnh. Sau y l 5 cu hi m cc ban nn tu tr
li v quyt jnh:
. Lnh vuc nghin cu ny c quan trong tiu th gi?
. tng nghin cu c th m rng v ng gp vo su nghip?
3. B mn ti ang lm vic c ph hop vi tng ca ti?
. tng c phn nh suy ngh ca ng nghip hin nay?
5. Ti am hiu v y vn trong lnh vuc ny, v lnh vuc no cn khai thc hay tm hiu
thm?
. Nghin cu ca ti c th lp vo khong trng tri thc?
. c nhiu nghin cu v ti cha, v ti c ng gp g thm?
. Thi im thch hop cho nghin cu?
. Nghin cu ca ti c gy tc ng trong chuyn ngnh?
. Trnh chuyn mn ca ti c ph hop vi muc tiu?
. Ti c k nng cn thit cho nghin cu?
. Ti c th gi theo ui du n?
3. Ti quyt tm vo du n?
. Ti c phong tin trong tay thuc hin?
5. Ti c ng nghip c chuyn mn hop tc?
Tm Iai, vit cong nghin cu l mt k nng rt quan trong ca mt nh khoa hoc. in nhn
manh l "rt quan trong (ch khng phi "quan trong). cc nc phong Ty, khi mt nh
khoa hoc c kh nng vit mt cong nghin cu, th l mt chng c v su trng thnh
ca nh khoa hoc, v l mt nc thang nh khoa hoc tr nn c lp.
Trc khi kt thc, ti mun k cc ban mt cu chuyn vui nhng hon ton c tht. Hm no,
gp anh ban [gi] ng nghip Melbourne, ti hi anh dao ny ra sao, anh th di ni ChD von
chin <u vi A c)ng nghin cu, Anh hi ti, v ti cng ni cng s phn. Anh y ci ni
sau khi xong ci "ull kighting ny chng ta s hop tc trong mt du n rt ho hng. Anh ban ti
xem vic vit vit cong nghin cu c ti tro nh l mt cuc <u "K; ti v anh y cng
ln u trng. C th c hai u tht bai, cng c th c hai u thnh cng, hoc chi mt
trong hai thnh cng. in ti tro qu tht l mt cuc u tranh. Khng chi u tranh trn mt
trn tng, m cn mt trn ch ngha! C ch l mt chuyn, nhng dng ch sao cho
thuyt phuc l mt k nng c ngha sng cn trong cuc u tranh xin ti tro.
in nhc lai rng cong nghin cu khng phi l khoa hoc, m l mt cch tip thj khoa hoc.
Tip thj bng ch ngha. Ghi nh im quan trong ny vit cong sao cho thuyt phuc
(ch khng phi "khoe) v tng xc sut oc ti tro. Chc cc ban may mn!
Ch thch:
. y l bi ti vit lai t workshop v vit cong nghin cu oc thuc hin ai hoc
Quc gia TPHCM (VN) vo nm . Trong workshop th chi c nhng slides, ch cha c
bi vit chi tit, v bi ny l mt bi "ti liu oc thm cho cc ban. Ti mun nhn co hi ny
by t lng bit on n cc ng nghip VN, c bit l Trung tm o tao v pht trin ngun
nhn luc (nay l Vin o tao quc t), tao iu kin thuc hin workshop, m theo cc hoc
vin l gip ch cho ho nhiu. Mt s cn "khoe xin oc ti tro v hoc bng nc ngoi.
in cm on cc ban hoc vin.
. Cc ban c th copy v phn phi bi ny cho cc ban khc (nu cn). Nhng ti chi mong
mun mt iu: ghi ngun. C nhiu khi ti thy mt s khng t bi vit v d liu ca mnh
oc s dung trong cc hi nghj (v c trn nhng trang bo) m tc gi khng ghi ngun, v
iu ny lm ti thy . hoi bun. Cng sc b ra rt nhiu, thu thp d liu, su tm nhng ca
th vj, ri rt rut v lao tm soan, m ngi ta s dung mt cch "v t, khng c n mt
ch ghi nhn ch cha ni g n cm on. Ni th thi, ch nu cc ban s dung v c kt qu
tch cuc l ti vui ri (khng cn cm on u).
3. Ngy / v / ny, nhn mt chuyn cng tc H Ni, ti s ni v cch vit cong
nghin cu Bnh vin Nng. Ti s c bi ging lin quan n nhng vn bn trong
bi ny. Nu cc ban c djp tham du th chc vui lm, v c nhiu co hi tho lun thm. Bi
ny tht su oc soan cho cc hoc vin Nng, v soan trong mt thi gian rt ngn (chi
ting ng h). V qu vi, nn chc cn nhiu thiu st. Nu cc ban pht hin sai st, xin gi
th cho ti bit v chinh sa sau ny. Ti hi vong s c djp quay lai b sung nhiu chi tit hon,
c bit l v du t VN (t cc ban).