Professional Documents
Culture Documents
Nguyn Vn Tun
GS H New South Wales, Sydney
i vi nhng ngi nh ti, vit cng nghin cu gn nh l mt ngh. 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 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.
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 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 th ng 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 niv 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 bitrt c 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.
b.
c.
d.
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 vitMt trong
nhng vn y t quan trng 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?
Mc tiu ca ti;
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
Vit Nam
M (NIH)
10
Mc tiu
Kt qu s khi
6-8
Phng php
10-20
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:
Cite trch dn: iu bt buc l nu dng d liu ca ng nghip th phi trch dn; v
Trong phn tm quan trng (significance), tc gi cn phi gii trnh rng cng trnh nghin
cu s c tc ng n:
Chuyn ngnh;
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.
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. InSpecific 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
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
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 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 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
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 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:
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:
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
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!
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 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