KHOA HOC KY THUAT THU Y - TAP XVIII - s 6 5 - 2011
THICH NGHI CAY CHUYEN CHUNG VIRUT DAI PV QUA TE BAO Nguyen Kim Dung, TrSn Thi My Dung.
Nguyin V5n ThUOng, Mai Thu Thao Vidn Pasteur Tp. HS Oil Minh
TOM TAT
Chung virut dai PV14 co ngu6n gtfc tCf chung PV13 cua Vifin Pasteur Paris, vtfn chi thich nghi tifim truydn qua nao ddng vSt. Vific thich nghi chung dfi' co th^ nhSn Ifin trong t^ bao la cin thifi't khi nghifin CUM phat trifin vacxin dai tir nudi tfi'bao dung cho ngudi va dflng vat d ntfdc ta.
Chung PV14/nao tho da dudc tifi'n hanh thich nghi ca'y chuyfin qua tfi' bao BHK-21 va Vero (WHO). Sau 4 lln c4'y chuyfin qua BHK-21, hifiu gia chung dat 10*'FFU/ml. CSy chuyin qua tfi' bao Vero 3 iSn tit PV17/BHK-21, hifiu gia virut dat 10*' FFU/ml. Khang nguyfin tvr chiing PV/BHK-21 CO tinh sinh miln dich va hifiu qua bao vfi chu6t nhat tr3ng ttft. 80 - 100% chutft co mijt khang thfi' du bao vfi {> 0,5 lU/ml) sau 2 va 4 tuln tifim khang nguyfin. 70 - 90% chudt s6ng sot sau khi dudc tifim 2 miii khang nguyfin cac lifiu 1/125 - 1/5 va thii thach vao nao 68 LD50 chiing CVS/nao chutft.
Titldtoa: Chiing virut dai PV va CVS; Vacxin dai tijf nu6i tfi' bao; Dap ling khang thfi; Hifiu qua bao vfi.
Adaptation of rabies virus PV strain in cell culture Nguyen Kim Dung, Tran Thj My Dung.
Nguyen Van Thi/ong, Mai Thu Thao SUMMARY
Rabies virus PV14 strain is rabbit-brain fixed rabies virus originally from PV13 strain of Pasteur Institute in Paris since 1992. Adaptation of this strain for research and development of cell-culture rabies vaccines in Vietnam is necessary. Rabies virus PV14/rabbit-brain was adapted in BHK-21 and Vero (WHO) cell culture. Virus titre of PV/BHK-21 strain reached 1 0 ' ' FFU/ml after 4 times passed in BHK-21. Following 3 Hmes passed in Vero (WHO) cell, titre of PV/Vero was 1 0 " FFU/ml. Antigen from PV/BHK-21 induced a good antibody response and protective effect. 80 — 100% mice got protective antibody level (>0.5 lU/ml) in 2 and 4 weeks. 70 — 90% mice were survived after 2 injections of antigen at 1/125 — 1/5 dilution and challenged in cerebral inoculation by 68 LDjj of rabies challenge virus strain (CVS).
Key KTorcb. Rabies virus PV and CVS sbrain; Cell culture rabies vaccine; Antibody response;
Protective effect.
KHOA HOC KY THUAT THU Y - TAP XVIII - s 6 5 - 2011 I. D ^ T VAN DE
Ta't ca cac chung virut dai dung lam vacxin cho ngtfdi va dflng vflt hifin nay d^u la con chau ciia m^t trong ba chung btf me duy nha't la chung nguyfin thuy Pasteur 1882, chiing FluryvaSAD[l].
Trong do, chiing PV13 ( Vifin Pasteur Tp.
HCM nhan tii Vifin Pasteur Paris n3m 1991) CO ngu6n gtfc tir chiing Pasteur 1882 [2], NSm 1992, chiing nay da dtfOc tifim truydn qua nao tho con va d6ng khO thanh chiing PV14. Tir do dtfOc sur dung dfi san xuS't vacxin dai diing cho ngtfdi cho dfi'n nSm 2006 tai Vifin Pasteur Tp. HCM cung nhtf 6 cac Vifin san xua't vacxin ciia Vifit Nam. Sau nSm 2007, khi vacxin tir nao chuflt sd sinh khtfng con dtfdc dimg d^ phong bfinh dai cho ngtfdi cf ntfdc ta, da CO mflt sfl" nghifin ctfu phat trifin vacxin dai dimg cho ngtfdi va dflng vat b3ng ky thuat nufli tfi' bao nham chii dflng ngu6n vacxin dai trong ntfdc. Do kho co thfi co chiing virut dai da thich nghi qua nufli ca'y tfi' bao tir cac nha sanxua'tvacxindaikhac, Ttf chiic Y tfi'thfi'gidi ciing khflng co chinh sach cung ling chiing san xua't vacxin dai, bfin canh do, ky thuat va di^u kifin tri^n khai nufli ca'y tfi' bao ra't thuan Idi tai Vifin Pasteur Tp. HCM, nhom nghifin Cliu da tifin hanh thich nghi ca'y chuyin chung PV14 qua nufli ca'y 2 dong tfi' bao Vero va BHK-21. Virut dai chung PV ca'y chuydn qua nufli ca'y tfi'bao BHK-21 (PV/BHK) sau khi bat boat da dtfdc ki^m tra tinh sinh mi§n djch va hifiu qua bao vfi chutft nhat trdng, so sanh vdi chiing CVS ca'y chuydn qua tfi' bao BHK-21 {CVS/BHK).
II VAT LIEU VA PHUONG PHAP NGHIEN Ctfu
2.1 Vat lifu
- Chung PVl^nao tho nhan dtfdc tir tifim truyin chung PV13 qua nao tho con nSm
1992. Chung dtf^c bao quan or -ZS^C dtfdi dang huySn djch nao 10% dflng khfl. Hifiu gia ciia chiing dat ID*-" LD50/ml khi c h u ^ dfl trfin chu0t.
Chung CV^HK-21 nhSn tir Vifin Pasteur Paris nSm 1992. Chiing nay da dtfdc thich nghi ca'y chuyin qua tfi'bao BHK-21 355 l^n. Sau 6 Mn ca'y chuyin thfim h Vifin Pasteur Tp. HCM, chiing dtfdc bao quan ii ddi cS'y 361. Hifin la chiing dimg cho thii nghifim ki^m tra hifiu gia khang th^ trung hoa virut dai trfin tfi' bao. Chiing nay ciing dtfdc nghifin ciiu dimg san xua't vacxin dai tir nufli tfi'bao dimg cho thii y.
- Tibio BHK-21 nhan tir Trung tflm ki^m Boat bfinh CDC Atlanta nSm 1998. Sau khi ca'y chuyin difim 4 lin, NgSn hang tfi'bao BHK-21 lam vific dtfdc bao quan d - 73''C. Cac tfi'bao la'y ra stf dung dtfdc ca'y chuyin thfim 2 — 3 Idn trtfdc khi thich nghi ca'y chuyin chiing PV14/ nao tho.
- Tibio Vera (WHO) ECACC 88020401 nhan dtfdc tir Ngin hang chiing gifl'ng chau Au (ECACC) nam 2005. Ngan hang tfi'bao gflc (MCB) Vero d ddi ca'y 137P va Ngan hang tfi' bao lam vific (WCB) Vero ii ddi ca'y 139P dtfdc bao quan -196*^:. Tfi'bao Vero d ddi ca'y 140P va 141P dtfdc diing dfi' ca'y chuyin tif chiing PV/BHK-21.
- Chudt nhit tring Swiss 14 — 16g ciia Vifin Pasteur Tp. HCM dtfdc dimg dfi' kifi'm tra tinh sinh miln djch va hifiu qua bao vfi ctia cac khang nguyfin tir chung PV/BHK-21 va CVS/BHK-21.
2.2 Fhtfong phap
- Thich nghi ciy chuyin chung PV14 qua nu6icaytibao BHK-21 va Vero
2 d'ng chung PV14/nao tho 10% dflng khfl dtfdc pha trong 2ml mfli trtfdng M199 1% huyfi't thanh bao thai bo (M199/1%FBS).
24
KHOA HOC KY T H U A T THU Y - T A P XVIII - s 6 5 - 2011 Sau khi iSc bang may trfin Vortex 600rpm/2
phiit, huy^n dich dtfdc ly tam 6000 vdng/phiit trong 2 phiit. Pha 1ml djch ntfi va 10 ml M199/1%FBS, loc 0,45Mm. Ca'y 0,2 ml dich qua loc vao mfli gifi'ng ciia phifi'n 12 gifi'ng da CO tham tfi' bao BHK-21 2 ngay.
Lang diu va ii 36,5°C trong 1 gid. Cho vao mfli gifi'ng 1 ml M199/1%FBS. U phifi'n trong tli a'm 36,5''C co 5%C02. Thay mfli trtfdng M199/1%FBS cho tS't ca cac gifi'ng vao ngay thii 7. Ngay thii 11, gat djch ntfi trong tiing gifi'ng vao 1 tfng Cryo day nhon. Ki^m tra stf hifin difin ciia virut dai trong dich gSt b^ng each ca'y 0,1 ml Ifin tham tfi' bao BHK-21 trong phifi'n 96 gifi'ng, thfim 0,1 ml M199/1%FBS va ii trong tii a'm 36,5''C c6 5%C02, sau 48 va 72 gid ctf dinh va nhuflm phifi'n bang thufl'c thtf mifin dich huynh quang phat hifin virut dai (Biorad Code 357- 2114). Trfln chung cac dich gSt co virut, chia 0,5 ml/fl'ng va bao quan d -73''C. Lfl virut nay dtfdc dat tfin la PV15/BHK-21. Tifi'n hanh ca'y chuyin chiing PV15/BHK-21 qua tfi' baa BHK-21 nufli trong cac chai 75cm^ va chai Roux 150 cm". Gat virut each 2 — 3 ngay tii ngay 2 dfi'n 14 va la'y mlu bao quan - 2 0 ^ cho dfi'n khi chua'n dfl. Ca'y chuyin qua BHK-21 3 Idn dfi' co cac chiing PV16/BHK-21; PV17/BHK-21 va PV18/BHK-21. TCr chiing PV17/BHK-21, ca'y chuyin qua tfi' bao Vero di c6 cac ifl chung PV18/Vero; PV19/Vero va PV20/yero. Cac chiing virut sau khi chua'n dfi dtfdc bao quan d -73''C. ChuS'n dfl virut cac lfl chiing trfin tfi' bao BHK-21 va phat hifin virut dai b^ng thufl'c thii miln dich huynh quang nhtf da nfiu trfin. Hifiu gia virut dtfdc tinh bang FFU/ml (Focus Fluorescent Unit). Chup anh quan sat hinh anh te" bao BHK-21 nhi^m virut dai cac chiing VP/BHK-21 va PV/Vero. So sanh vdi hinh anh ciia virut dai chiing CVS/BHK-21.
- Kiim tra tinh sinh miin d/ch vi hiiu qua bao vi chudt nhit tring ciia khang nguyfin tir chiing PV/BHK-21, so sanh vdi khang nguyfin hi chiing CVS/BHK-21: Hifiu gia virut trtfdc ba't boat ciia khang nguyfin PV/BHK-21 la IC'^FFU/ml va khang nguyfin CVS/BHK-21 la lO'-^FFU/ml. Ba't boat cac huy§n djch virut bling Betapropiolactone 1/4000, pha chfi' trong cha't tfn dinh va 10%
cha't btf h-d mi^n djch Pet gel A (Seppic). Kifi'm tra b'nli smh miln dich bSng each tifim dtfdi da mfli chuOt 0,5 ml khang nguyfin. Mfii lfi tifim cho 15 chuflt. Cac chuflt nay dtfdc la'y mau sau 1, 2 va •] luAn. La'y mau tim, tach, ba't boat huyfi't thanh 56'*C/30 phiit va bao quan -20°C cho dfin khi c h u ^ dfl. Khang thfi' trung hda dtfdc xac diiili b3ng phtfdng phap FAVN.
Kifi'm tra hifiu qua bao vfi dtfdc xac dinh bang phtfdng phap NIH: kliang nguyfin pha 1/5;
1/25 va 1/125, mfli dfl pha tifim 0,5ml/chu6t/6 bimg, 2 mui each nhau 1 tudn.
Tuan thii 3 tifim vao nao chuflt lieu thtf thach 10 ~ 100LD50/0,03ml chiing CVS (Challenge virus strain). Theo doi chuflt 14 ngay va ghi nhan stf Itfdng chuflt chfi't do bfinh dai.
III. KET QUA V A T H A O LUAN 3.1 Chung PV thich nghi c^y chuyfi'n qua tfi' bao Vero va BHK-21:
Sau 4 Idn cSy chuyin qua ttf bao BHK-21, bifiu gia virut ciia chiing PV da tang tti 10^^
FFU/ml Ifin 10"' FFU/m! trong dich gSt nufli tfi' bao (Dfl thi 1). Tlidi difi'm stf Itfdng virut giai phong vao mfli trtfdng cung nhanh hdn.
Tir lan thich nghi l/BHK-21 dfi'n lin 3/BHK- 21, stf Itfdng vtfut cao nha't vao ngay thii 11- 12, d ^ lan ca'y thii 4/BHK-21, ngay thii 10, hifiu gia virut da dat 10*^' FFU/ml. 6 tfi' bao Vero, sau 3 Idn ca'y chuyin tir chiing PV17/BHK-21, dfin ddi ca'y 6/Vero, Itfdng virut dat dtfdc cao nha't vao ngay 8 va duy tri cao dfi'n ngay 12 (dfl thi 1).
KHOA HOC KY THUAT THtJ Y - TAP XVIII - s 6 5 - 2011
8 -|
7 6
iUFFL
S i
1 1
1 -)
1
i / m i K
6 7 ^ • '
1 1 1 1
i l l
1 1 L
^.0 r..3-
2/UI IK .l/liHK 4/iil IK - 5/Vcro 6A/cro
4/Vcro • PV/BHK-21
i ^"in Ciiy chuyen tir chiuig VP 14/nao Ilxi D P W e r o
Do thf 1. Hieu gia virut chung PV cJy chuyen qua te bio Vero va BHK-21 Quiin s.il dtfoi kinh hiOn vi, cac tc b.'io c6
virut bdt huyiili quting n<im rai r<ii., Uicli rdi i-ihau dfi'i vdi Cii P V / B H K - 2 1 (liinh la, lb) v.i P V / V e r o (hinli 2a, 2b), DiC-u dc'- dw thay tinh lay t r u y i n tha'p ciia c h u n g PV gitfa c i c te bao so vdi chiing C V S / B H K - 2 1 (hinli 3) vtfn la
c h u n g da difdc thich n g h i ca'y chuyfin nhieu lan q u a tO bao. N g a v ca d m a t d o cao (hinh 2a) chuiij; PV c u n g it tao c i c d a m cum te hao nhiC'm \'irul bat liu\'nh q u a n g nhtf cac dam c u m h u v i i h q u a n g Idn cua chiing CVS/BHK- 21 (hinh 3),
Hinh 1. Chung PV/BHK-21 sau 1 ISn (la) vi3I3'n (lb) ciy chuyin tUtPVl^iao tho. Huyen dich virut git sau 12 ngiy ciy trin tham tibio BHK-21; inh chup sau 72 (la) va 48 (lb) gid
26
KHOA HOC KY THUAT THU Y - TAP XVIII - s 6 5 - 2011
(2a) (2h)
Hinh 2. Chiing r\/\'fio -.m 3Lin i.h iliuycn tii I'V/BIIh-JI. Ilu\4n dich litatg.ft s.tu 12 ngay-dpph.i 10' {2.il \.i1(i (2h) >.Jy lien lluini te b.m BHK-21; jtih chup sau 4H gia
Hinh 3 Chung CVS361/BHK-21 ciy chuyin tti chung CVS35^BHK-21. Huyin dich virut git sau 4 ngiy ddpha Iff' ciy trin tham tibio BHK-21; anh chup sau 48gid 3 ^ Tinh sinh mifin djch va hieu qua bao vfi
chuflt nhit tr^ng cua chung PV qua nufli cS'y tfi^bio BHK-21
Hifiu gia khang th^ trung hda (HGKTTH) h nhom chuflt tifim khang nguyfin PV/BHK-21 xua't hifin sdm d 4 / 5 chuflt sau 2 hidn. HGKTTH dat trung binh 4,7 lU/ml.
Dfi'n tudn thir 4, HGKTTH trung binh dat 15,4 lU/ml, 5/5 chuflt d i u co miic khang
thfi' bao vfi > 0,5 lU/ml. Nhom chuflt tifim khang nguyfin CVS/BHK-21, HGKTTH xudt hifin mufln hdn trong thi nghifim nay. Sau 2 tudn chtfa co khang thfi' ci 5/5 chuflt. Dfi'n tudn thii 4, mdi cd 3/5 chufit cd khang thfi' > 0,5 lU/ml. Do mflt chuflt co miie khang thfi' ra't eao Ifin tdi 81,3 lU/ml nfin HGKTTH trung binh ca nhom dat 17,5 lU/ml (Bangl).
KHOA HOC KY THUAT T H O Y - TAP XVIII - s 6 5 - 2011
Bangl. Tinlt sinh miin djch cua khing nguydn tii chung VP/BHK-21 so v6i tu; chiing CVS/BHK-21
Khing nguydn
PV/BHK-21 CVS/BHK-21
Ty id chudt cd ididng thi bio vd>0.5 iU/ml S Hidu gii khing Ihi Imng binh (iU/mi) 1 l u i n
Tyl«
0/5 (0%) 0/5 (0%)
Hi$u gid IU/ml 0,2 ±0.1
0
2 t u i n
TylS 4/5 (80%)
0/5 (0%) Hi$u gid
IU/ml 4.7 ±6.1
0
4 tuin T y l * 5/5(100%)
3/5 (60%) Hi$u gid
IU/ml 15,4 ±11.8 17,5 ±35,7
Khi thii thach vdi chung thii thach 90% chuflt 0 cac dfl pha khang nguyfin 1/125 C:VS/nao chuflt lilu 68 LD50/0,D3inl, chuijt — 1/5. Ty lfi nay i chuflt tifim khang nguyfin tifim khang nguyfin PV/BHK-21 bao vfi 70 — CVS/BHK-21 la 10 —100% (Cang 2).
Bang 2. Hidu qua bao vd chudt nhit tring cija khang nguydn tit chung PV/BHK-21 so vii ta Chung CV^HK-21
Khing nguydn
PV/BHK-21 CVS/BHK-21
Ty id chudt sdng/ ddpha khing nguydn 1/5
9/10(90%) 9/9(100%)
1/25 9/10(90%) 7/10(70%)
1/125 7/10 (70%) 1/10(10%)
Lidu bdo vd 50% chudt
Logic ED50 -2.7 -1.65
THAO LUAN
Mac dii CO ra't nhi^u loai vacxin dai diing cho ngtfdi va dflng vSt nhtfng ta't ca cac chung san xua't vacxin d^u co ngufln gflc tit 3 chung chinh Pasteur (PV), Flury va SAD [1]. Thi du cac chung virut dai PM (Pittman Moore) va Kissling cd ngufln gfl'c tii nao cua 1 bd difin 6 Phap nam 1882 (chiang PV). Chung virut dai PM dixdc dimg di san xuS't vacxin tvf tfi' bao ltfdng b0i ngtfdi (HDCV), vacxin tir tfi' bao Vero tinh chfi' (PVRV) va vacxin phfli vjt tinh chfi' (PDEV) [3]. Chung Pitman-Moore (PM) ciing dtfdc xem nhtf chung virut dai Pasteur ctf djnh ca'y chuydn qua nao tho PV -11 [2].
Chung virut dai ctf d)nh CVS cung cd ngutfn
gflc tir chung PV nhimg dtfdc ctf djnh cS'y chuydn qua nao chuflt rdi sau do thich nghi ca'y chuydn qua BHK-21. Cd mflt stf khac bifit v^ cae quyfi't dinh khang nguyfin trfin protein G cua 2 chung PV va CVS khi nhan difin bang khang thfi' ddn dong Mabs-G (4]. Trong nghifin ciiu [4], chiing PV cd 23 quyfi't dinh khang nguyfin dtfdc Mabs-G nhan ra trong khi chiing CVS chi co 17 vi tri. So sanh doan ngoai mang (1-439) cua protein G, chiing PV cd 4 vi tri (37,158, 247, 319) dtfdc glycosyl hoa trong khi chiing CVS chi co 3 (37, 204 va 319).
Tuy nhifin ca 2 chung d^u gitfng nhau la co Arginine d vi tri 333 (vj tri gSy dflc th^n kinh).
Protein G ciia virut dai la khang nguyfin kich tttf'ch tao khang th^ trung hoa virut va bao vfi
28
K H O A H O C K Y T H U A T T H O Y - TAP XVIII - s 6 5 - 2 0 1 1
dflng vat k h i thtf t h a c h bling d t f d n g tifim n a o va ngoai vi. T r o n g nghifin ciiu n a y , c h u n g PV d u m d i dtfdc thich n g h i c^'y c h u y d n q u a tfi'bao BHK-21 nhtfng d a cho tha'y nhtfng tfu difi'm v^
tinh sinh mifin dich v a hifiu q u a bao vfi dflng vat so v d i chiing C V S / B H K - 2 1 . T h e o [5], cac chiing Vtfut d a i thich n g h i ca'y c h u y d n q u a tfi' bao thtfdng n h a n rS't n h a n h v a g a y d a p ling mifin dich m a n h c h o cd th^. Kfi't q u a q u a n sat chiing C V S / B H K - 2 1 cho tha'y v i r u t lay i i h i l m n h a n h tii tfi'bao n a y s a n g t ^ ' b a o khac, tao cac d a m c u m tfi' b a o chiia v i r u t bit m a u h u y n h q u a n g Idn s a u 48 gid. Mfli d a m c u m co tlifi' Ifin tdi 30 - 40 tfi'bao t r o n g k h i chiing P V / B H K - 2 1 chl tao n h i i n g d a m c u m h u y n h q u a n g n h o k h o a n g 5 - 1 0 tfi'bao.
K E T L U A N
Chiing v i r u t d a i PV cfl' d i n h , tifim t r u y i n qua n a o tho d a dtfdc thich n g h i cS'y c h u y d n ttianh cflng q u a te b a o BHK-21 va Vero. Hifiu gia vtfut ciia P V / B H K - 2 1 d a t c a o nha't 10*' F F U / m l d d d i ca'y 4, lfl P V 1 8 / B H K - 2 1 . Hifiu gia virut ciia P V / V e r o c u n g d a t tdi lO'*'^
F F U / m l s a u 3 l l n ca'y c h u y i n tii P V 1 7 / B H K - 21. K h a n g nguyfin P V / B H K - 2 1 g a y d a p ling mifin dich n h a n h s a u 2 t u a n va 80 -100% chuflt cd d a p ling k h a n g thfi' dii bao vfi s a u 2 va 4 t u l n . K h a n g nguyfin V P / B H K - 2 1 ciing bao v e tfl't 70 - 90% chuflt tifim e a c l i l u 1 / 1 2 5 - 1 / 5
khi thtf thach v a o n a o chiing v i r u t d a i cd dflc Itfc v d i 68 LDso. C h i i n g P V / B H K - 2 1 va P V / V e r o d a dtfdc thich ling q u a tfi'bao nufli CO thfi chpn l a m chiing san xua't vacxin dai tir nufli tfi'bao cho ngtfdi va dflng vat.
TAI L I E U T H A M K H A O t Rhodes A. ).: Strains of rabies virus
available for preparaUon of Sylvatic rabies vaccines with special reference to vaccines prepared in cell culture. Can. Vet. J. 2 (Ausu.ft 1981)2:262. 266.
2. WHO: Report of German Green Cross/ WHO workshop on monoclonal antibody in rabies diagnosis and research. WHO/Rab.
Res./S4.20.
3. Moore SM, Ricke TA, Davis RD, Briggs DJ.
The influence of homologous vs.
heterologous challenge virus strains on the serological test results of rabies virus neutralizing assays. Bioiogicals. 2005; 33(4):
269-276. [PubMed: 16168666],
t . Smith J.S., King A.A: Monoclonal antibodies for the identification of rabies and non-rabies lyssaviruses. World Health Organization, Geneva, Laboratory techniques in rabies 4*
edition, 145 -155
5 Dielzschold B, Li J, Faber M, Schnell M.:
Concepts in the pathogenesis of rabies.
Future Virol 2C08Sep;3(5):487^90.
Thdng bao b6' nhifm Tong bien tap tap chi
T a p chl K H K T thii y r a m 3 t b a n doc Kr t h a n g 8 / 1 9 9 3 , dfi'n n a y d a p h a t h a n h t a p XVIII.
GS.TS D a o T r o n g D a t , nguyfin Vifin trtfcfng Vifin thii y, n g u y e n Chii tich Hfii thu y VN, ngtfdi s a n g l a p T a p chi v a giii chiic Tfl'ng bifin t a p T a p chi tir d o dfin n a y .
Nay d o tudi cao (80 tufl'i), thfi' theo nguyfin v o n g CLUI GS Dat dtfdc nghi d i l u h a n h tap chi, Ban thtfdng v u Hfli thii y Vifit N a m d a thfl'ng nliS't dfi' GS thfli giil chiic TBT va bfi' nhifim GS.TS Dau Ngoc H a o , Pho chii tich Hfli, P h o TBT tap chi giii chiic TBT kfi' tir 1 / 8 / 2 0 1 1 .
Hfli thii y Vifit N a m trSn t r o n g d a n h gia cao stf d d n g g d p to Idn ciia GS D a t trong sufl't thdi gian q u a t r o n g vific p h a t t r i ^ n T a p chi va d l nghi GS tifi'p tuc giiip d d T a p chi trong vific bifin t a p v a d o n g g d p x a y dtfng T a p chi n g a y c a n g viing m a n h dfi xiing d a n g v d i l o n g m o n g m o i ciia a n h chi e m t r o n g n g a n h . Kinh chiic GS lufln k h o e m a n h va h a n h p h u c .
Hoi thu y Viet Nam
