Y HQC VIET NAM THANG 7 - SO 1/2011
3. Mike Lean, Martin Wiseman (2008):
Malnutrition in hospitals, 2008, 336: 290.
4. Cinda S. Chima, Kathy Barco, Marci L.A.
Dewitt, M.L.A Dewitt, Michelle Maeda, J.Carlos Teran, Kevin D. Mullen (1997):
Relationship of nutrition status to length of stay, hospital costs, and discharge status of patients
hospitalized in the medicine service. Joumal of the American Dietetic Association, 975-978.
5. Leonor Rodriguez, Elsa Ceriantes and Rode Ortiz (2011): Malnutrition and gastrointestinal and respiratory infections in children: A pubic health problem. Int J. Res. Publk: Health, 1174-1205.
6. Martin TR (1987): The relatkxTship between malnutrition and lung infections. Qin Chest Med, 359-72.
EBNA-1 CUA VIRUS EPSTEIN-BARR 0 MOT SO MAU NPC TAI HA NOI Dime XAC DjNH THUOC PHAN TYP V-VAL
^Hoang Van Manh va ^Le Thanh Hda
T 6 M TAT
EBNA-1 lien quan mat thiet den ung thU vom mui hong va tang nhiem virus. Phan tich da hinh tai vj tri add amin 487 cho phep xac dinh 5 phan typ trong do c6 V-val (V: Variant; v: valine). Mau benh pham NPC ciia benh nhan Ha Ndi cd phan typ V-val hay khdng la van de ran lam sdng td. Chung tdi thUc hien phat hi|n Vci xac dinh phan typ EBV bing phUdng phap phan tich d§c diem phan tir EBNA-1 tren 4 mau benh pham sinh thiet tir benh nhan m§c NPC nhap vien tai Ha Noi.
Cac mau dUdc tach DNA, thUc hien PCR, thu nhan doan gen EBNA-1 (~0,77 kb); giai trinh tU, so sanh;
phan tich acid amin EBNA-1 va nhan dinh da hinh tai vj tri 487 de xac djnh phan typ. Ket qua cho thay, trong sd 4 mau NPC ciia benh nhan Ha Noi phat hien phan typ: V-val, chula phat hien V-leu va V-pro va cac phSn t^p khac. NhU vay, V-val ton tai trong so 4 mau NPC va vi^c sang loc phan tuf xac dinh phan typ EBNA- 1 \h can thiet trong djnh hUdng nghien cufu djch te va phdng ehdng NPC tai Vi§t nam.
TCr khoa: EBV, NPC, EBNA-1, PCK phan typ, V-val.
SUMMARY
DETERMINATION OF V-VAL SUBTYPE FROM EBNA-1 OF EPSTEIN-BARR VIRUS FROM NPC
PATIENTS ADMITTED I N HA NOI
EBNA-1 is associated with tumorigenesis and persistent latent infection of EBV. Based on the site- variation at amino acid 487, EBNA-1 is dassified into 5 subtypes induding subtype V-val (V: Variant; vai:
valine). It should be elucidated that how many and which subtypes of EBNA-1 In the Hanoi's EBV isolates are. Detection and subtyping EBV collected in Hanoi has been canied out by molecular analysis of EBNA-1 of 4 biopsy isolates from the NPC patients. DNA extraction, PCR amplification, EBNA-1 fragment (~0.77 kb), sequendng, comparative analysis, annotation of
^Dai hgc Thai Nguyen; •'\^ien Cong nghe sinh hgc Phan bl^ khoa hpc: GS.TS. Dai Duy Ban
amino acids, polymorphism determination at the aa 487 position were undertaken for subtyping. /\s results, unique V-val subtype is detected in 4 isolates of Hanoi patients; not yet V-leu and V-pro as previously determined. It can be conduded that subtype V-val detected in 4 EBV isolates facilitating molecular screening for essential subtyping of EBNA-1 in terms of epidemiological and preventive studies of the NPC in Vietnam.
Key words: EBV, NPC, EBNA-1, PCR, subtype, V-val.
I. O^T V A N o f
Ung t h u vdm mui hong (NPC, nasopharyngeal carcinoma), Burkitt lymphoma, benh Hodgkin T-lymphoma dUde col la cd vai trd xuc tien eiia Epstein-Barr virus (EBV) (Young, Rickinson, 2004). Protein EBNA-1 (Epstein-Bar Nuclear Antigen 1) gom 641 acid amin (aa), la loai cd mat trong tat ca moi dang tien trien khdi u ciia EBV, xuc tien qua trinh tang nhiem anh hudng den sU nhan len ciia he gen, sao chep va phan chia te bao (Murray, Young, 2001).
Polypeptide EBNA-1 cd mot viing gom nhieu motif lap glycine/alanine (GAR) tir aa 90 den aa 327 (Mai et al., 2007; 2010) va cac amino acid vj tri tir 459 den 607 dau tan ciing C ed vai trd trung gian cho DNA bam dinh va hinh thanh phan tir kep (dimerization) (Wu et al., 2002). Da hinh tai vj tri add amin 487 eho phep xac djnh phan typ V-val (V: variant), lien quan den luu hanh EBV trong md, mau va djch hong (Bhatia et al., 1996). Theo nghien cii'u gan day, EBV phan lap d viing Nam Trung Qudc va cac mau EBV phan lap tai Viet Nam chii yeu la V-val, ddng vai b d
49
Y HOC VIET NAM THANG 7 - SO 1/2011
quan trpng tang cUdng sao chep ddi vdi EBNA-1 (Mai et al., 2007; 2010; Le Thanh Hda va cs, 2010).
Ben canh nghien cii'u lam sang, benh tich, tien trien benh, EBV phan lap tai Viet Nam cung da dUdc nghien cifu ve dac diem sinh hpc phan tir, trong dd cd phan tich gen EBNA-1 va LMP-1 (Do et al., 2008; Nguyen Oinh Phuc, Le Thanh Hda, 2010). Tuy nhien, viec xac djnh phan typ (subtyping) dUa tren bien ddi acid amin 487 eiia EBNA-1 chUa dUdc die cap nhieu tai Viet Nam (Le Thanh Hda va es, 2010). Muc tieu eiia nghien ciru nay la lam sang td them trong cac mau NPC ciia benh nhan tai Ha Ndi nhti'ng nam 2000 thudc phan typ nao.
Trong bai bao nay, chung tdi phan tich da hinh tai vj tri acid amin 487 ciia EBNA-1 de xac djnh cac phan typ Val-variant d 4 mau benh pham cua benh nhan ung thu vdm hpng (NPC) nhap vien tai Ha Npi.
II. NGUYEN VAT LIEU VA PHUONG P H A P
NGHIEN COU
2.1. Mau benh pham va phUdng phap xu* ly Mau tUdi thu bang phUdng phap sinh thiet tir benh nhan ung thU vdm hpng (NPC), nhap vien nhii'ng nam 2000 (gpi chung la EBV(2000)), ky hieu rieng tiTng chiing la: EB17; EB18; EB19;
EB20. Mau dUdc bao quan d -20°C.
DNA tdng sd dUdc tach chiet suf dung bd kit ciia hang BIONEER (Han Qudc). Tdm tat: nghien mau thanh huyen djch; cho proteinase K, ii va cho RNase. Cho dung mdi GC, ii d 70°C, roi cho isopropanol, ly tam de thu djch. Chuyen len cdt lpc, ly tam, loai bd djch dUdi. Cho dung mdi W l , ly tam, bd djch dUdi. Cho dung mdi W2, ly tam, bd djch dudi. Chuyen cdt sang dng Eppendorf mdi, cho djch tach EL, ly tam, thu djch ben dUdi la DNA tdng sd ciia mau, bao quan d -20°C cho den khi sir dung.
2.2. ThUc hien PCR, giai trinh tU, xuf ly sd lieu
Doan gen EBNA-1 (khoang 0,77 kb) thu dUdc bing PCR vdi khudn la DNA tdng sd, sir dung cap moi: EBKIF: 5' GTCATCAT6\TCCGGGTCTC 3' va EBVR: 5' CGATTGAGGGCGTCTCCTAAC 3', mdt ta trUdc day (Le Thanh Hda va cs, 2010). Giai trinh tU trUc tiep san pham PCR sau khi tinh sach de thu nhan chuoi nucleotide doan gen EBNA-1 va xii ly bang chUdng trinh SeqEdl.03, sap xep bang AssemblyLIGNl.9 va MacVeetor8.2 (Aecelrys
Inc.). Thanh phan amino acid suy dien bang each sir dung bd ma ciia vi sinh vat bac thap (vi khuan, bacterial code) cd trong Ngan hang gen va so sanh ddi chieu bang chUdng trinh GENEDOC2.5. Tim amino acid tai vj tri 487 de xae djnh mdt trong 5 phan typ (Bhatia et al., 1996).
III. KET QUA NGHIEN COU
3.1. Thu nhan chuoi gen EBNA-1
Vdi cap moi EBKIF - EBVR, doan gen khoang 0,77 kb dUdc thu nhan, chiing tdi tien hanh tinh sach san pham PCR va giai trinh tU trUc tiep, ket qua trinh bay d Hinh 1. San pham gen EBNA-1 ddn bang, chat lUdng tdt, chung tdi giai trinh true tiep sir dung mdi EBKIF.
Hinh 1 . Dien di kiem tra san pham PCR doan gen EBNA-1 tren thach agarose 1%. Ghi chu: EB17;
EB18; EB19 EB20: san pham PCR ciia cac mau nghien ciru; DC(+): doi chifng dUOng (co khudn EBNA-1); DC (-): doi chifng am (kh6ng co khu6n EBNA-1).
3.2. Xac djnh phan typ EBV qua phan tich Vj tri aa 487 cua EBNA-1
Vi tri acid amin 487 dUdc sir dung de so sanh ddi chieu EBNA-1 ciia 4 mau EBV(2000) thu nhan tir benh nhan NPC nhap vien tai Ha Ndi (EB17, EB18, EB19, EB20) va mdt so chiing Viet Nam mdi thu nhan gan day la chung HP6 (Hal Phdng); chiing EB9 (Ha Npi) dai dien benh nhan nhap vien tai mien Bac va chiing MT7 (Mien Trung) dai dien mien Trung (Hinh 2).
Ket qua so sanh trinh bay d Hinh 2 cho thay, tai vj tri 487, 4 chimg EBV ciia Ha Ndi chira 1 loai acid amin, dd la V (valine), tir dd xae djnh thudc phan typ: V-val (Variant valine).
Ket qua xac djnh phan typ cua 7 chiing, bao gom ca 4 chiing nghien ciTu va 3 chiing tham chieu ciia Viet Nam. dUdc liet ke d Bang 1. Bang 1 cung cho thay chi tiet, ca 4 mau nghien ciru thudc V-val (EB17, EB18, EB19, EB20); mau EB9 (Ha Ndi) thupc V-val; mau Mien trung thudc P-thr (MT7); mau Hai Phdng thudc mau P-ala (HP6).
50
Y HQC VigT NAM THANG 7 -SOI /2011
EB17 EB18 : EB19 EB20 : EB9 tun HPe :
* 2C
aaafiaBeaaBSaff SBB
*
^^aMiaaga| ^^Si
40 *
^ ^ ^ ^ ^ jIHlSjatjKj KCX
60 agnas^j
*
isji^gp
a a 4 8 7 80 t *
^ ^ r a v ^ ^ raSRf
100
Hinh 2. So sanh mpt phan trinh tU acid amin EBNA-1 cua 7 chiing, gom 4 chiing dang nghien cifu la EB17; EB- 18; EB-19;^EB2j); chiing EB9 ciia Ha Noi, chiing MT7 ciia mien trung va chiing HP6 ciia Hai phong. Vj tri aa 487 dUOc chi dan bang mui ten; aa d vj tri 487 ciia cac chiing EBV so sanh dUdc dong khung doc.
Bang 1. Ket qua xae <^nh phian typ ciia cac chiing EBV qua phan tfch acfcl amin tai vj tri 487 ciia EBNA-1 TT
1 2 3 4 5 6 7
Chung EB17 EB18 EB19 EB20 EB9 MT7 HP6
Nam phan lap 2000 2000 2000 2000 2000 2009 2009
33 4 8 7 V V V V V T A
Ket qua xac djnh phan typ
V-val V-val V-val V-val V-val P-thr P-ala
IV. BAN LUiBlN
Ket qua xac djnh acid amin vj tri aa 487 ciia 4 chiing EBV Ha Npi cho thay deu thudc phan typ V-val; cac mau nay dddc phan lap nam 2000.
Theo nghien cii\i cua tac gia Le Thanh Hda va cs (2010), cd khi phan tich 22 mau nghien cult thu thap cac nam 2000 va gan day (2008-2009), 14 mau CO phan typ V-val; trong do tat ca cac mau nSm 2000 deu thupc phan typ V-val (Le Thanh Hoa va cs, 2010). NhU vay cho den nay, ehUa phat hien phan t^p khac ngoai V-val d cac mau NPC thu nhan trUdc nam 2000. Mot dieu lUu y la EBV cd dac tinh da hinh bien ddi giii'a cac chiing phan lap d cac viing dja ly thude Nam va Ddng Nam chau A (Wang et al., 20ici) va nghien cii\j ciia Mai et al (2007; 2010) chiiYig minh cac chiing EBV thudc V-val d viing Nam Trung Qudc, CO kha nang tang cUdng sao chep va cd vai trd quan trpng trong tien trien NPC. Xac djnh dUdc phan typ (V-val) trong sd mau EBV chiing tdi nghien cihj (EB17, EB18, EB19, EB20) va so sanh (EB9) ciia Ha Npi, giup chiing tdi khang djnh cac mau nghien cufU nam 2000 <feu thupc V-val, cdn cac mau MT7, HP6 nam 2009 xuat hien thupc P- ala va P-thr. Tuy so mau chUa nhieu, nhUhg nghien CU\J ciia chung tdi cho thay EBV Viet Nam, ve mat djch te hpc, cd bj anh hUdng ciia vet djch te EBV tir Quang Ddng Nam Trung Qudc tdi Ddng Nam chau A.
V. KET LUAN
Phan t^p V-val dUdc phat hien ton tai trong sd 4 mau NPC cua Ha Ndi phan lap nam 2000 khi so sanh vdi cac mau gan day. Phan typ V-val ndi trpi trong tat ca cac mau, dUdc xac nhan cd vai trd trong tien trien ung thU (tumorigenesis) cua NPC va phan bd dja ly viing Nam va Odng Nam A. Sang lpc phan tuf xac djnh phan typ EBNA-1 de djnh hirdng phdng chong NPC la can thiet tai Viet Nam.
LOI CAM ON: Chiing toi chan thanh cam dn ho trd kinh phi cua Phdng Mien dich hgc, Vien Cong nghe sinh hoc va Bg Khoa hgc va Cong nghe thong qua de tai KClO.31/06-10 de thu'c Men cong trinh nay.
TAI LIEU THAM KHAO
1. Bhatia K, R3j A, Guitierrez MI, Judde JG, Spangler G, Venkatesh H and Magrath IT (1996). Variation in the sequence of Epstein Barr virus nudear antigen 1 in normal peripfieral blood lymphocytes and in Burkitt's lymphomas.
OJCB^ene 13:177-181.
Do NV, Ingemar E, Phi PT, Jenny A, Chinh TT, Zeng Y and Hu L (2008). A major EBNAl variant from Asian EBV isolates shows enhanced transcriptional activity compared to prototype B95.8. Virus Res 132(1-2): 15-24.
Le Thanh Hoa, Vu Thi TICT, Hoang Thj Minh Chau, Le Th3nh Ha, Nguyen Oinh Phuc (2010). Xac dinh 3 phan typ P^la, P*r va V-val oJa EBNA-1 d EBV phan lap tai Viet Nam. T^ dvN^Nen ahi K/wc70(5): 8-12.
2.
3.
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Y HOC VIET NAM T H A N G 7 - SO 1/2011
4. Mai S3, Ooka T, Li DJ, Zeng MS, Jiang RC, Ju XJ, Zh3ng RH, Chen SP 3nd Zeng YX (2007).
Functional advantage of NPC-related V-val subtype of Epstein-Barr virus nuclear antigen 1 compared with prototype in epithelial cell line.
Oncoic^y Reports, 17:141-146.
5. Mai SJ, Xie D, Hu3ng YF, Wsng FW, Liao YJ, Deng HX, Liu WJ, Hua WF and Zeng YX (2010). The enhanced transcriptional activity of the V-val subtype of Epstein-Barr virus nuclear
antigen 1 in epithelial cell lines. Oncol Rep 23(5): 1417-24.
6. Murray PG, Young LS (2001). Epstein-Ban^
virus infection: basis of malignancy and potential for therapy. Expert Rev Moi Med, 3(28): 1-20.
7. Nguyen Dinh Phuc, Le Thanh Hoa (2010).
Nghien cifu cau true gen LMP-1 ciia cac chiing EBV gay ung thU vom mui hong d Viet Nam. Tap chi Yhgc VietNam, 1: 40-47.
DANH GIA SUTIEN TRIEN CAN TH| SAU 1 NAM ft HOC SINH LftP 6 TRU&NG THCS
CAT L I N HHA NOI
T6M T&T
D$t van de: Can thj 6 hgc sinh nUde ta ngay cang gia t3ng ve ty le mac va mire do tien trien, can nghien cufu sU tien trien ciia can thj de cd cac giai phap ng3n chan. Muc tieu.{l) Danh gia sU tien trien aia can thi d hoc sinh khoi 6 trudng PTCS Cat tinh. Ha Noi trong 1 n§m hoc (2) Khao sat mpt sd yeu td lien quan.
Phu'dng phap: Nghien ciru c3t ngang, theo doi tien ciru ti-en 225 hoc sinh ldp 6 trUdng THCS Cat Linh.
Ket qua: Sau 1 nam can thi kh6ng thay ddi d 30,0%
sd mit, tien trien cham 20,0%, tien trien trung binh (TB) 40,5%, tien trien nhanh 6,8%, tien trien rat nhanh la 2,6%, mifc dp tien trien 0,4 D ± 0,6D/nam.
Cac yeu td lien quan: Can thi d tre ni? tien trien TB 0,2D/n3m; co bd/me b| can tien trien TB 0,4D/n3m; sir dung mat nhin gan tren 8 gid /ngay tien trien TB 0,4D/n§m; hoc Idp chuyen tang TB 0,4D/nam; kh6ng hoat dong ngoai tr&i >2 gid/ngay tang TB 0,3D/nam. Ket luan: Can thf d hgc sinh Idp 6 tiWdng Cat Linh tien tii&i TB 0,4 D ± 0,6D/nam. Can co cac giai phap dong bo de han che sU tien trien nhanh can thj d hoc sinh.
Tit khoa: Tien trien can thi hoc sinh
SUMMARY
ASSESSMENT OF MYOPIA PROGRESSION AFTER ONE YEAR IN THE 6 GRADE PUPILS OF THE SECONDARY CATLINH SCHOOL IN HANOI IN 2010
Background: Myopia in the school children in Vietnam has been increased not only in the prevalence but also in progression so that it is necessary to research to have appropriate preventive solution A i m s ; l . To evaluate myopia progression after one year in 6 * grade pupils of the CatLinh School in Hanoi.
2. To describe some risk factors related to myopia progression. Methodoloov; Cross-sectional
Nguyen Chi Dung*
prescriptive study with one year follow up on 225 children aged from 11 to 12 years old. Results; After one year Myopia is not progressed in 30% of eyes, but it slowly progressing in 20,0% of eyes, moderately in 40,5% and rapidly in 6,8% and very rapidly in 2,6% of eyes with 0,4D ± 0,6D/year averagely. Risk factors related: myopia is progressing in the female children with 0,2 D/year averagely, in those who have the parents suffered from RE with 0,4D/year, in the children who are using close vision more than 4 hour/day with 0,4D/year, in the children of the spedal dass with 0,4D/year and in those who dont have the regular outside activities with 0,3D/year averagely.
Conclusion; Myopia of the 6 * grade pupils of the CatLinh School in Hanoi, in 2010 is progressing with 0,4 D/year averagely. It is necessary to have comprehensive solutions in order to restrict a rapid progression of myopia in children
Key words: myopia progression in children
I.
OAT VANof
Hien nay ty le tre em hpc sinh mac TKX, dac biet la can thj rat cao va phd bien d mii'c tir 35- 42% d cac thanh pho Idn. Cupc song hien dai, chUdng trinh hpc tap nang ne vdi thdi glan nhin gan nhieu trong ngay la dieu kien thuan Idi khien ti le mac TKX, dac biet la can thj ngay mpt tang nhanh. Mat khac, can thj cung ngay cang tien trien nhanh hdn khien tre phai tang sd kinh nhieu hdn gay anh hUdng nghiem trpng den thj lUc va chat lUdng cupc sdng ciia tre mac TKX.
Tren the gidi da cd nhieu nghien cii'u ve sU tien trien ciia tat khiic xa ndi chung va can thj ndi rieng. Nghien cii'u cua Goss DA va Cox VD d tre
* Benh vien Mit Trung u'dng 52
