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n\paperw11900\paperh16840\pard\sb0\sl-240{\bkmkstart Pg1}{\bkmkend Pg1}\par\pard
\ql \li3836\sb0\sl-241\slmult0 \par\pard\ql\li3836\sb0\sl-241\slmult0 \par\pard\
ql\li3836\sb0\sl-241\slmult0 \par\pard\ql\li3836\sb0\sl-241\slmult0 \par\pard\ql
\li3836\sb57\sl-241\slmult0 \up0 \expndtw-3\charscalex100 \ul0\nosupersub\cf15\f
16\fs21 2012 International Conference on Bioscience, Biochemistry and Bioinforma
tics \par\pard\ql \li5621\sb1\sl-239\slmult0 \up0 \expndtw-3\charscalex100 IPCBE
E vol. 1(2012) (2012)IACSIT Press, Singapoore \par\pard\ql \li1996\sb0\sl-368\
slmult0 \par\pard\ql\li1996\sb0\sl-368\slmult0 \par\pard\ql\li1996\sb0\sl-368\sl
mult0 \par\pard\ql\li1996\sb0\sl-368\slmult0 \par\pard\ql\li1996\sb3\sl-368\slmu
lt0 \up0 \expndtw0\charscalex97 \ul0\nosupersub\cf1\f2\fs32 Gene Expression Stud
y of Breast Cancer Cell in Response \par\pard\ql \li4117\sb12\sl-368\slmult0 \up
0 \expndtw0\charscalex97 to Aloe Emodin Treatment \par\pard\ql \li1560\sb0\sl-27

6\slmult0 \par\pard\ql\li1560\sb72\sl-276\slmult0 \up0 \expndtw-4\charscalex100

\ul0\nosupersub\cf2\f3\fs24 Indah Mohd Amin\ul0\super\cf3\f4\fs23 1\ul0\nosupers
ub\cf2\f3\fs24 , Siti H Sheikh Abdul Kadir\ul0\super\cf3\f4\fs23 1\ul0\nosupersu
b\cf2\f3\fs24 , Nik Mohd Mazuan Nik Mohd Rosdy\ul0\super\cf3\f4\fs23 2\ul0\nosup
ersub\cf2\f3\fs24 and \par\pard\ql \li4927\sb154\sl-264\slmult0 \up0 \expndtw-3
\charscalex100 Narimah AH Hasani\ul0\super\cf3\f4\fs23 1 \par\pard\ql \li2450\sb
146\sl-253\slmult0 \up0 \expndtw-3\charscalex100 \ul0\super\cf4\f5\fs21 1\ul0\no
supersub\cf5\f6\fs22 Institute for Medical Molecular Biotechnology (IMMB), Facu
lty of Medicine, \par\pard\ql \li3842\sb1\sl-237\slmult0 \up0 \expndtw-4\charsca
lex100 Universiti Teknologi MARA (UiTM) Malaysia. \par\pard\ql \li2839\sb70\sl-2
53\slmult0 \up0 \expndtw-4\charscalex100 \ul0\super\cf4\f5\fs21 2\ul0\nosupersub
\cf5\f6\fs22 Faculty of Dentistry, Universiti Teknologi MARA (UiTM) Malaysia. \
par\pard\qj \li1558\sb0\sl-230\slmult0 \par\pard\qj\li1558\ri1387\sb56\sl-230\sl
mult0\fi0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf6\f7\fs24 Abstract.\ul
0\nosupersub\cf7\f8\fs20 Breast cancer is the most common and cause of cancer d
eath among Malaysian women. Despite \up0 \expndtw0\charscalex100 many therapeuti
c advances in the treatment of this cancer, overall the incidence rate is still
unacceptable. \up0 \expndtw-3\charscalex100 Mortality that results from the brea
st cancer is still alarming. There is emerging attention in the use of natural \
up0 \expndtw0\charscalex100 products as a new chemopreventive agent. Aloe emodin
(Ae), a natural active compound present in Aloe \up0 \expndtw0\charscalex102 Ve
ra\ul0\nosupersub\cf8\f9\fs20 \ul0\nosupersub\cf7\f8\fs20 leaves has been repor
ted to have several medicinal properties including antioxidant, antiviral, and \
up0 \expndtw-3\charscalex100 anticancer activities. This paper provides a litera
ture review of Ae with an insight on the experimental design \up0 \expndtw0\char
scalex100 to be used. This study is using microarray as a tool, and it is a hope
that the transcriptional profiles might \up0 \expndtw0\charscalex100 give a new
understanding on how the Ae compound affect the gene alteration. Protein study
also will be \up0 \expndtw-1\charscalex100 carried out to integrate with molecul
ar findings that might provide basic rational for in vivo study or can be \up0 \
expndtw-3\charscalex100 extended into clinical setting. The outcomes of this stu
dy may explore widened possibilities of the medicinal \up0 \expndtw-3\charscalex
100 properties of Ae in other types of cancer. \par\pard\ql \li1558\sb141\sl-241
\slmult0 \up0 \expndtw-3\charscalex100 \ul0\nosupersub\cf6\f7\fs24 Keywords:\ul0
\nosupersub\cf7\f8\fs20 breast cancer, aloe emodin, anticancer. \par\pard\ql \l
i1133\sb313\sl-322\slmult0 \up0 \expndtw-6\charscalex100 \ul0\nosupersub\cf9\f10
\fs28 1.\ul0\nosupersub\cf10\f11\fs28 \ul0\nosupersub\cf9\f10\fs28 Introductio
n \par\pard\qj \li1134\ri951\sb93\sl-280\slmult0\fi357 \up0 \expndtw-3\charscale
x100 \ul0\nosupersub\cf5\f6\fs22 In recent years, breast cancer becomes the comm
onest cause of cancer related death in Asia [1]. National \line \up0 \expndtw-1\
charscalex100 Cancer Registry of Malaysia provides an age standardized incidence
rate (ASR) for 2004 of about 46.2 per \line \up0 \expndtw0\charscalex103 100,00
0 women. The rate however differs among three main races whereby the Chinese is
the highest, \line \up0 \expndtw-1\charscalex100 followed by Indians and Malays
[2]. Extensive studies in the last decade using microarray technology have \line
\up0 \expndtw0\charscalex100 provided better understanding of human cancer part
icularly in breast cancer. It has significantly showed a \line \up0 \expndtw-1\c
harscalex100 way to predict prognosis, treatment response and helping in making
the clinical decision [3]. Therefore, we \line \up0 \expndtw-3\charscalex100 exp
loit the use of this tool to identify the effect of Ae in breast cancer cell lin
es. Proteomic study will also be \line \up0 \expndtw-3\charscalex100 carried out
to further enhance our understanding from the underlying molecular mechanism to
the functional \line \up0 \expndtw0\charscalex102 stage of the cell. To date, n
o studies have been conducted to correlate between the effect of Ae in breast \l
ine \up0 \expndtw0\charscalex100 cancer cells with the variation in gene express
ion as well as the regulatory protein involved. Preliminary \line \up0 \expndtw3\charscalex100 study [4] revealed that Ae has antiproliferative effect in breas
t cancer cell line. \par\pard\ql \li1133\sb186\sl-322\slmult0 \up0 \expndtw-6\ch
arscalex100 \ul0\nosupersub\cf9\f10\fs28 2.\ul0\nosupersub\cf10\f11\fs28 \ul0\n
osupersub\cf9\f10\fs28 Literature Review \par\pard\ql \li1133\sb187\sl-287\slmu

lt0 \up0 \expndtw-2\charscalex100 2.1.\ul0\nosupersub\cf10\f11\fs28 \ul0\nosupe

rsub\cf6\f7\fs24 Aloe Vera \par\pard\qj \li1133\ri953\sb0\sl-280\slmult0\fi357
\up0 \expndtw0\charscalex100 \ul0\nosupersub\cf5\f6\fs22 Aloe is a genus and one
of the widely known species is Aloe Vera or also known as \ul0\nosupersub\cf11\
f12\fs22 Aloe barbadensis \line \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf
5\f6\fs22 Miller\ul0\nosupersub\cf11\f12\fs22 .\ul0\nosupersub\cf5\f6\fs22 Ae i
s the well-known anthraquinone active compound that can be found in some species
of Aloe [5]. \line \up0 \expndtw-2\charscalex100 Literature of different compon
ents found in Aloe species and their medicinal properties is shown in Table 1. \
par\pard\ql \li2901\sb81\sl-230\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosup
ersub\cf7\f8\fs20 Table 1: Literature of different components and medicinal prop
erties of Aloe Vera \par\pard\li1242\sb0\sl-184\slmult0\par\pard\li1242\sb97\sl184\slmult0\fi0\tx5308\tx9446 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf13
\f14\fs16 Components of Aloe Vera\tab \up0 \expndtw-2\charscalex100 Medicinal pr
operties\tab \up0 \expndtw-2\charscalex100 Author(s)\par\pard\ql \li5862\sb124\s
l-207\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf14\f15\fs18 41
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87507c45ff00a4c2ff00c15fff00f0f5fecb3ffd0715faff0045007e407fc3a83e22ff00d2617fe0
afff00f87aff00659ffe838a3fe1d41f117fe930bff057ff00fc3d7fb2cfff0041c57ebfd1401f90
1ff0ea0f88bff4985ff82bff00fe1ebfd967ff00a0e28ff87507c45ffa4c2ffc15ff00ff000f5fec
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c3a83e22ff00d2617fe0afff00f87aff00659ffe838a3fe1d41f117fe930bff057ff00fc3d7fb2cf
ff0041c57ebfd1401f901ff0ea0f88bff4985ff82bff00fe1ebfd967ff00a0e28ff87507c45ffa4c
2ffc15ff00ff000f5fecb3ff00d0715faff45007e407fc3a83e22ffd2617fe0aff00ff0087aff659
ff00e838a3fe1d41f117fe930bff00057fff00c3d7fb2cff00f41c57ebfd1401f901ff000ea0f88b
ff004985ff0082bfff00e1ebfd967ffa0e28ff0087507c45ff00a4c2ff00c15fff00f0f5fecb3ffd
0715faff0045007e407fc3a83e22ff00d2617fe0afff00f87aff00659ffe838a3fe1d41f117fe930
bff057ff00fc3d7fb2cfff0041c57ebfd1401f901ff0ea0f88bff4985ff82bff00fe1ebfd967ff00
a0e28ff87507c45ffa4c2ffc15ff00ff000f5fecb3ff00d0715faff45007e407fc3a83e22ffd2617
fe0aff00ff0087aff659ff00e838a3fe1d41f117fe930bff00057fff00c3d7fb2cff00f41c57ebfd
1401f901ff000ea0f88bff004985ff0082bfff00e1ebfd967ffa0e28ff0087507c45ff00a4c2ff00
c15fff00f0f5fecb3ffd0715faff0045007e407fc3a83e22ff00d2617fe0afff00f87aff00659ffe
838a3fe1d41f117fe930bff057ff00fc3d7fb2cfff0041c57ebfd1401f901ff0ea0f88bff4985ff8
2bff00fe1ebfd967ff00a0e28ff87507c45ffa4c2ffc15ff00ff000f5fecb3ff00d0715faff45007
e407fc3a83e22ffd2617fe0aff00ff0087aff659ff00e838a3fe1d41f117fe930bff00057fff00c3
d7fb2cff00f41c57ebfd1401f901ff000ea0f88bff004985ff0082bfff00e1ebfd967ffa0e28ff00
87507c45ff00a4c2ff00c15fff00f0f5fecb3ffd0715faff0045007e407fc3a83e22ff00d2617fe0
afff00f87aff00659ffe838a3fe1d41f117fe930bff057ff00fc3d7fb2cfff0041c57ebfd1401f90
1ff0ea0f88bff4985ff82bff00fe1ebfd967ff00a0e28ff87507c45ffa4c2ffc15ff00ff000f5fec
b3ff00d0715faff45007e407fc3a83e22ffd2617fe0aff00ff0087aff659ff00e838a3fe1d41f117
fe930bff00057fff00c3d7fb2cff00f41c57ebfd1401f901ff000ea0f88bff004985ff0082bfff00
e1ebfd967ffa0e2ab5cffc1257e205dac2b2ff00c1623fe0b10820b9b7bb436df1e3f665b26696da
412c693359fec7f035c5b33002e2cae0cb67751e62ba82689990fec451401f901ff0ea0f88bff498
5ff82bff00fe1ebfd967ff00a0e28ff87507c45ffa4c2ffc15ff00ff000f5fecb3ff00d0715faff4
5007e407fc3a83e22ffd2617fe0aff00ff0087aff659ff00e838a3fe1d41f117fe930bff00057fff
00c3d7fb2cff00f41c57ebfd1401ce783b40baf0a7847c2de17bef14788fc6f7de1bf0e687a05e78
d3c63268d2f8bbc5f75a3e996ba75c78a3c532f87745f0e787a4f11ebf35b3eadae49a1787b41d19
f53bbba6d2f45d2ec8c1630747451400514514005145140051451400514514005145140051451400
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51451400514514005145140051451400514514005145140051451400514514005145140051451400
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51451400514514005145140051451400514514005145140051451400514514005145140051451400
5145140051451400514514005145140051451400514514005145140051451401ffd9}}}}}\pard\s

b0\sl-240{\bkmkstart Pg2}{\bkmkend Pg2}\par\pard\li1241\sb0\sl-184\slmult0\par\p

ard\li1241\sb0\sl-184\slmult0\par\pard\li1241\sb0\sl-184\slmult0\par\pard\li1241
\sb0\sl-184\slmult0\par\pard\li1241\sb171\sl-184\slmult0\fi0\tx5103\tx9687 \up0
\expndtw-1\charscalex100 \ul0\nosupersub\cf3\f4\fs16 High molecular fraction (ba
rbaloin & polysaccharide)\tab \up0 \expndtw-1\charscalex100 \u8226?\ul0\nosupers
ub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 hypoglycemic effect on type 2 diab
etis mellitus\tab \dn3 \expndtw-1\charscalex100 [6]\par\pard\li1241\sb56\sl-184\
slmult0\fi0\tx5103\tx9687 \up0 \expndtw-1\charscalex100 Phytosterols\tab \up0 \e
xpndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3
\f4\fs16 hypoglycemic effect on type 2 diabetis mellitus\tab \dn3 \expndtw-1\cha
rscalex100 [7]\par\pard\li1241\sb56\sl-184\slmult0\fi0\tx5103\tx9687 \up0 \expnd
tw-1\charscalex100 Polysaccharides\tab \up0 \expndtw-1\charscalex100 \u8226?\ul0
\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 immunity & antioxidant\ta
b \dn3 \expndtw-1\charscalex100 [8]\par\pard\li1241\sb56\sl-184\slmult0\fi0\tx51
03\tx9687 \up0 \expndtw-1\charscalex100 Protein (14 kDa)\tab \up0 \expndtw-1\cha
rscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 ant
ifungal & anti-inflammatory\tab \dn3 \expndtw-1\charscalex100 [9]\par\pard\li124
1\sb56\sl-184\slmult0\fi0\tx5103\tx9646 \up0 \expndtw-1\charscalex100 Phenolic c
ompounds\tab \up0 \expndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16
\ul0\nosupersub\cf3\f4\fs16 protective & sealing effect against infection after
wounding\tab \dn3 \expndtw-1\charscalex100 [10]\par\pard\li1241\sb56\sl-184\slm
ult0\fi0\tx5103\tx9473 \up0 \expndtw-1\charscalex100 Aloesin\tab \up0 \expndtw-1
\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16
antioxidant, anti-inflammatory & anticancer\tab \dn3 \expndtw-1\charscalex100
[11], [12]\par\pard\li1241\sb56\sl-184\slmult0\fi0\tx5103\tx9646 \up0 \expndtw1\charscalex100 Barbaloin\tab \up0 \expndtw-1\charscalex100 \u8226?\ul0\nosupers
ub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 anticancer\tab \dn3 \expndtw-1\c
harscalex100 [12]\par\pard\li1241\sb56\sl-184\slmult0\fi0\tx5103\tx9646 \up0 \ex
pndtw-1\charscalex100 Glycoprotein\tab \up0 \expndtw-1\charscalex100 \u8226?\ul0
\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 wound healing\tab \dn3 \e
xpndtw-1\charscalex100 [13]\par\pard\qj \li1134\sb0\sl-280\slmult0 \par\pard\qj\
li1134\ri951\sb18\sl-280\slmult0\fi357 \up0 \expndtw0\charscalex100 \ul0\nosuper
sub\cf5\f6\fs22 Numerous studies also reported on medicinal properties derived f
rom different extracts of Aloe Vera. \up0 \expndtw0\charscalex105 The pharmacolo
gical elements of this species are studied extensively via in vivo and in vitro
as well. \up0 \expndtw-4\charscalex100 Summary of the literature is shown in Tab
le 2. \par\pard\ql \li3391\sb102\sl-230\slmult0 \up0 \expndtw-3\charscalex100 \u
l0\nosupersub\cf7\f8\fs20 Table 2: Literature on medicinal properties extract of
Aloe Vera \par\pard\li3085\sb0\sl-184\slmult0\par\pard\li3085\sb92\sl-184\slmul
t0\fi0\tx4785\tx6487\tx7958 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf13\f
14\fs16 Type of extract\tab \up0 \expndtw-2\charscalex100 Medicinal properties\t
ab \up0 \expndtw-2\charscalex100 Type of study\tab \up0 \expndtw-2\charscalex100
Author(s)\par\pard\li3085\sb0\sl-180\slmult0\fi0\tx4785\tx6488\tx8160 \up0 \exp
ndtw-2\charscalex100 \ul0\nosupersub\cf3\f4\fs16 Pulp\tab \up0 \expndtw-2\charsc
alex100 detoxification agent\tab \up0 \expndtw-2\charscalex100 in vivo\tab \up0
\expndtw-2\charscalex100 [14]\par\pard\li3085\sb2\sl-184\slmult0\fi0\tx4784\tx64
88\tx8160 \up0 \expndtw-2\charscalex100 Pulp & liquid fraction\tab \up0 \expndtw
-2\charscalex100 antifungal\tab \up0 \expndtw-2\charscalex100 in vitro\tab \up0
\expndtw-2\charscalex100 [15]\par\pard\li3085\sb0\sl-184\slmult0\fi0\tx4785\tx64
87\tx8159 \up0 \expndtw-2\charscalex100 Pulp\tab \up0 \expndtw-2\charscalex100 a
nticancer\tab \up0 \expndtw-2\charscalex100 in vivo\tab \up0 \expndtw-2\charscal
ex100 [16]\par\pard\li3085\sb0\sl-183\slmult0\fi0\tx4785\tx6489\tx7988 \up0 \exp
ndtw-2\charscalex100 Gel\tab \up0 \expndtw-2\charscalex100 wound healing\tab \up
0 \expndtw-2\charscalex100 in vivo\tab \up0 \expndtw-2\charscalex100 [17], [18]\
par\pard\li3085\sb2\sl-184\slmult0\fi0\tx4785\tx6487\tx8160 \up0 \expndtw-2\char
scalex100 Gel\tab \up0 \expndtw-2\charscalex100 antimicrobial\tab \up0 \expndtw2\charscalex100 in vitro\tab \up0 \expndtw-2\charscalex100 [19]\par\pard\li3085\
sb0\sl-184\slmult0\fi0\tx4786\tx6487\tx8006 \up0 \expndtw-2\charscalex100 Gel\ta
b \up0 \expndtw-2\charscalex100 anti-inflammatory\tab \up0 \expndtw-2\charscalex
100 in vitro, in vivo\tab \up0 \expndtw-2\charscalex100 [20],[21]\par\pard\li308

5\sb0\sl-183\slmult0\fi0\tx4786\tx6489\tx8161 \up0 \expndtw-2\charscalex100 Aeri

al parts\tab \up0 \expndtw-2\charscalex100 hepatoprotective\tab \up0 \expndtw-2\
charscalex100 in vivo\tab \up0 \expndtw-2\charscalex100 [22]\par\pard\ql \li1134
\sb200\sl-287\slmult0 \up0 \expndtw-3\charscalex100 \ul0\nosupersub\cf9\f10\fs28
2.2.\ul0\nosupersub\cf10\f11\fs28 \ul0\nosupersub\cf6\f7\fs24 Aloe Emodin \pa
r\pard\qj \li1134\ri951\sb0\sl-280\slmult0\fi357 \up0 \expndtw-3\charscalex100 \
ul0\nosupersub\cf5\f6\fs22 Various studies conducted have revealed that Ae can i
nduce cell death to various cancer cell lines. Table \line \up0 \expndtw-3\chars
calex100 3 provides the literature on the anticancer properties of Ae from diffe
rent type of human cancer cell lines. \par\pard\ql \li3270\sb101\sl-230\slmult0
\up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf7\f8\fs20 Table 3: Literature on
various mechanism of anticancer properties of AE \par\pard\li1242\sb0\sl-184\sl
mult0\par\pard\li1242\sb84\sl-184\slmult0\fi0\tx2660\tx4360 \up0 \expndtw-2\char
scalex100 \ul0\nosupersub\cf13\f14\fs16 Type of cancer\tab \up0 \expndtw-2\cha
rscalex100 Type human cell line\tab \up0 \expndtw-2\charscalex100 Various mechan
ism of anticancer properties of AE\par\pard\li1242\sb0\sl-181\slmult0\fi0\tx2660
\tx4360\tx10356 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf3\f4\fs16 Cervic
al\tab \up0 \expndtw-2\charscalex100 KB\tab \up0 \expndtw-2\charscalex100 \u8226
?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 induces in vitro G\
ul0\sub\cf18\f19\fs15 2\ul0\nosupersub\cf3\f4\fs16 /M arrest and activates alkal
ine phosphatase.\tab \up0 \expndtw-2\charscalex100 [23]\par\pard\li1242\sb11\sl184\slmult0\fi1418\tx4360\tx10355 \up0 \expndtw-1\charscalex100 HeLa\tab \dn2 \e
xpndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3
\f4\fs16 involves G\ul0\sub\cf18\f19\fs15 2\ul0\nosupersub\cf3\f4\fs16 /M arres
t and induce differentiation\tab \up0 \expndtw-1\charscalex100 [24]\par\pard\li1
242\sb12\sl-184\slmult0\fi0\tx2660\tx4360\tx10355 \up0 \expndtw-1\charscalex100
Lung\tab \up0 \expndtw-1\charscalex100 H460\tab \dn2 \expndtw0\charscalex100 \u8
226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 photosensitized
AE-induced anoikis associated with protein expression of \u945?-actinin &\tab \u
p0 \expndtw-1\charscalex100 [25]\par\pard\li2660\sb12\sl-184\slmult0\fi1859 \up0
\expndtw-1\charscalex100 mitogen-activated protein (MAP) kinase members.\par\pa
rd\li2660\sb1\sl-184\slmult0\fi0\tx4360\tx10355 \up0 \expndtw-1\charscalex100 H4
60\tab \dn2 \expndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\
nosupersub\cf3\f4\fs16 induces apoptosis through involvement of modulation
of cAMP-dependent protein\tab \up0 \expndtw-1\charscalex100 [26]\par\pard\li1
242\sb10\sl-184\slmult0\fi3278 \up0 \expndtw-1\charscalex100 kinase, protein kin
ase C, Bcl-2, caspase-3 and p38 protein expression.\par\pard\li1242\sb1\sl-184\s
lmult0\fi1418\tx4360\tx10355 \up0 \expndtw-1\charscalex100 CH27 & H460\tab \dn2
\expndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\c
f3\f4\fs16 induces apoptosis through involvement of protein kinase C\tab \up0 \
expndtw-1\charscalex100 [27]\par\pard\li1242\sb11\sl-184\slmult0\fi0\tx2660\tx43
60\tx10355 \up0 \expndtw-1\charscalex100 Gastric/ Stomach\tab \up0 \expndtw-1\ch
arscalex100 AGS & NCI-N87\tab \up0 \expndtw-1\charscalex100 \u8226?\ul0\nosupers
ub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 releases apoptosis-inducing fac
tor & cytochrome C from mitochondria, activates\tab \up0 \expndtw-1\charsc
alex100 [28]\par\pard\ql \li4520\sb10\sl-184\slmult0 \up0 \expndtw-3\charscalex1
00 caspase-3 that lead to nuclear shrinkage & apoptosis. \par\pard\li2660\sb2\sl
-184\slmult0\fi0\tx4360\tx10355 \up0 \expndtw-1\charscalex100 MGC-803 & SGC-\tab
\up0 \expndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupe
rsub\cf3\f4\fs16 involves in suppression of c-myc expression\tab \up0 \expndtw1\charscalex100 [29]\par\pard\ql \li2660\sb1\sl-171\slmult0 \up0 \expndtw-3\char
scalex100 7901 \par\pard\li2660\sb12\sl-184\slmult0\fi0\tx4394\tx10356 \up0 \exp
ndtw0\charscalex100 MGC-803\tab \dn2 \expndtw0\charscalex100 \u8226?\ul0\nosuper
sub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 induces growth inhibitory effect
with an increase in S phase and alkaline phosphatase\tab \up0 \expndtw0\charscal
ex100 [30]\par\pard\ql \li4536\sb3\sl-184\slmult0 \up0 \expndtw-3\charscalex100
activity repression \par\pard\li1242\sb9\sl-184\slmult0\fi0\tx2661\tx4360\tx1035
6 \up0 \expndtw-1\charscalex100 Nasopharynx\tab \up0 \expndtw-1\charscalex100 NP
C-TW 039 & NPC-\tab \dn2 \expndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f1
7\fs16 \ul0\nosupersub\cf3\f4\fs16 induces G\ul0\sub\cf18\f19\fs15 2\ul0\nosup

ersub\cf3\f4\fs16 /M phase arrest by increase cyclin B1 level bound to Cdc2\tab

\up0 \expndtw-1\charscalex100 [31]\par\pard\ql \li2660\sb1\sl-163\slmult0 \up0 \
expndtw-3\charscalex100 TW 076 \par\pard\li1242\sb20\sl-184\slmult0\fi0\tx2660\t
x4361\tx10356 \up0 \expndtw-1\charscalex100 Liver\tab \up0 \expndtw-1\charscalex
100 Hep G2 & Hep 3B\tab \up0 \expndtw0\charscalex100 \u8226?\ul0\nosupersub\cf16
\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 has antiproliferative activity throug
h p53-dependent and p21-dependent apoptotic\tab \up0 \expndtw-1\charscalex10
0 [32]\par\pard\li2660\sb12\sl-184\slmult0\fi1859 \up0 \expndtw0\charscalex100 p
athway\par\pard\li2660\sb1\sl-184\slmult0\fi0\tx4361\tx10356 \up0 \expndtw0\char
scalex100 Hep G2\tab \up0 \expndtw0\charscalex100 \u8226?\ul0\nosupersub\cf16\f1
7\fs16 \ul0\nosupersub\cf3\f4\fs16 affects multiple proteins associated
with oxidative stress, cell cycle arrest,\tab \up0 \expndtw0\charsca
lex100 [33]\par\pard\li1242\sb10\sl-184\slmult0\fi3278 \up0 \expndtw-1\charscale
x100 antimetastasis, and hepatitis C virus replication\par\pard\li1242\sb1\sl-18
4\slmult0\fi0\tx2661\tx4361\tx10356 \up0 \expndtw-1\charscalex100 Brain\tab \up0
\expndtw-1\charscalex100 SVG & U-373MG\tab \up0 \expndtw-1\charscalex100 \u8226
?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 modulates PKC isozy
mes, inhibits proliferation & induces apoptosis\tab \up0 \expndtw-1\charscalex10
0 [34]\par\pard\li1242\sb12\sl-184\slmult0\fi0\tx2660\tx4361\tx10356 \up0 \expnd
tw-1\charscalex100 Blood- leukemia\tab \up0 \expndtw-1\charscalex100 HL-60\tab \
dn2 \expndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupers
ub\cf3\f4\fs16 induces in vitro G\ul0\sub\cf18\f19\fs15 2\ul0\nosupersub\cf3\f4
\fs16 /M arrest of cell cycle\tab \up0 \expndtw-1\charscalex100 [35]\par\pard\li
1242\sb11\sl-184\slmult0\fi0\tx2661\tx4361\tx10356 \up0 \expndtw-1\charscalex100
Bladder\tab \up0 \expndtw-1\charscalex100 T24\tab \dn2 \expndtw-1\charscalex100
\u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\f4\fs16 induces apop
tosis through the p53 dependent apoptotic pathway\tab \up0 \expndtw-1\charscalex
100 [36]\par\pard\li1242\sb11\sl-184\slmult0\fi0\tx2660\tx4361\tx10356 \up0 \exp
ndtw-1\charscalex100 Tongue\tab \up0 \expndtw-1\charscalex100 SCC-4\tab \up0 \ex
pndtw-1\charscalex100 \u8226?\ul0\nosupersub\cf16\f17\fs16 \ul0\nosupersub\cf3\
f4\fs16 induces cell death through S-phase arrest and apoptosis\tab \up0 \expnd
tw-1\charscalex100 [37]\par\pard\ql \li1134\sb176\sl-322\slmult0 \up0 \expndtw-6
\charscalex100 \ul0\nosupersub\cf9\f10\fs28 3.\ul0\nosupersub\cf10\f11\fs28 \ul
0\nosupersub\cf9\f10\fs28 Research Framework \par\pard\qj \li1134\ri952\sb113\s
l-280\slmult0\fi357 \up0 \expndtw0\charscalex103 \ul0\nosupersub\cf5\f6\fs22 It
was previously found that Ae has significantly showed antiproliferative activity
in human breast \line \up0 \expndtw-2\charscalex100 cancer cell line, MCF-7. Th
erefore, this study aimed to screen the genes and proteins that may play a role
in \line \up0 \expndtw0\charscalex100 Ae-suppressed proliferation of breast canc
er cells. Further understanding and identifying the role of these \line \up0 \ex
pndtw-1\charscalex100 genes and proteins will enable us to understand th
e regulation of its metabolic activity. This study \line \up0 \expndtw-2\
charscalex100 hypothesized that Ae regulates the genes and proteins in the suppr
ession of breast cancer cells proliferation. \line \up0 \expndtw-2\charscalex100
The following figure 1 represents the research framework and the explanation of
each of the phases. \par\pard\ql \li5862\sb0\sl-207\slmult0 \par\pard\ql\li5862
\sb0\sl-207\slmult0 \par\pard\ql\li5862\sb0\sl-207\slmult0 \par\pard\ql\li5862\s
b0\sl-207\slmult0 \par\pard\ql\li5862\sb153\sl-207\slmult0 \up0 \expndtw-2\chars
calex100 \ul0\nosupersub\cf14\f15\fs18 42 \par\pard\sect\sectd\fs24\paperw11900\
paperh16840{\shp
{\*\shpinst\shpleft0\shptop0\shpright11900\shpbottom16840\shpwr3\shpfblwtxt0\shp
z-10000\shplid2
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d15f007fc2b9ff0082a6ff00d1e47ec01ff8ad3fda2bff00a6c547fc2b9ff82a6ffd1e47ec01ff00
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b0\sl-240{\bkmkstart Pg3}{\bkmkend Pg3}\par\pard\ql \li5799\sb0\sl-184\slmult0 \
par\pard\ql\li5799\sb0\sl-184\slmult0 \par\pard\ql\li5799\sb0\sl-184\slmult0 \pa
r\pard\ql\li5799\sb0\sl-184\slmult0 \par\pard\ql\li5799\sb0\sl-184\slmult0 \par\
pard\ql\li5799\sb0\sl-184\slmult0 \par\pard\ql\li5799\sb104\sl-184\slmult0 \up0

\expndtw-3\charscalex100 \ul0\nosupersub\cf19\f20\fs16 Oligonucleotide\par\pard\

sect\sectd\sbknone\cols2\colno1\colw5167\colsr60\colno2\colw6533\colsr160\ql \li
4665\sb74\sl-184\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf3\f4\fs
16 Gene\par\pard\ql \li2642\sb1\sl-150\slmult0\tx4541 \up0 \expndtw-2\charscalex
100 Cell Culture\tab \up0 \expndtw-2\charscalex100 Profiling\par\pard\ql \li2302
\sb70\sl-184\slmult0\tx3225 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf19\f
20\fs16 Normal\tab \up0 \expndtw-2\charscalex100 Breast\par\pard\ql \li2302\sb1\
sl-183\slmult0 \up0 \expndtw-7\charscalex94 Breast
\ul0\nosupersub\cf3\f4\f
s16
vs \ul0\nosupersub\cf19\f20\fs16
Cancer\par\pard\ql \li2302\sb4\sl-184
\slmult0\tx3206\tx4586 \up0 \expndtw-2\charscalex100 Cell Line\tab \up0 \expndtw
-2\charscalex100 Cell Line\tab \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf3
\f4\fs16 Protein\par\pard\ql \li4540\sb61\sl-184\slmult0 \up0 \expndtw-2\charsca
lex100 Profiling\par\pard\ql \li2715\sb0\sl-184\slmult0 \par\pard\ql \li2715\sb5
1\sl-184\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf13\f14\fs16 PHA
SE I\par\pard\column \ql \li752\sb21\sl-184\slmult0 \up0 \expndtw-2\charscalex10
0 \ul0\nosupersub\cf19\f20\fs16 microarray\par\pard\ql \li695\sb142\sl-184\slmul
t0\tx3303 \up0 \expndtw-2\charscalex100 Validation with\tab \up0 \expndtw-2\char
scalex100 \ul0\nosupersub\cf3\f4\fs16 Data\par\pard\ql \li575\sb38\sl-144\slmult
0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf19\f20\fs16 real time RT-PCR\p
ar\pard\ql \li3170\sb0\sl-144\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosuper
sub\cf3\f4\fs16 Analysis\par\pard\ql \li5227\sb0\sl-184\slmult0 \par\pard\ql \li
665\sb12\sl-184\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf19\f20\f
s16 2-D eletrophoresis\par\pard\ql \li5227\sb0\sl-184\slmult0 \par\pard\ql \li20
\sb81\sl-184\slmult0\tx3146 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf13\f
14\fs16 PHASE II\tab \up0 \expndtw-2\charscalex100 PHASE III \par\pard\sect\sect
d\sbknone \ql \li4789\sb121\sl-230\slmult0 \up0 \expndtw-3\charscalex100 \ul0\no
supersub\cf7\f8\fs20 Fig. 1: Research Framework. \par\pard\ql \li1134\sb223\sl-2
87\slmult0 \up0 \expndtw-1\charscalex100 \ul0\nosupersub\cf9\f10\fs28 3.1.\ul0\n
osupersub\cf10\f11\fs28 \ul0\nosupersub\cf6\f7\fs24 Phase I \par\pard\qj \li11
33\ri953\sb0\sl-280\slmult0\fi357 \up0 \expndtw0\charscalex100 \ul0\nosupersub\c
f5\f6\fs22 Breast cancer cell line (MCF-7) and normal breast cell line (MCF-10A)
will be cultured in complete \up0 \expndtw0\charscalex100 DMEM and DMEM/HAM\u82
17?s F-12 medias (FLOWLAB, Sydney, Australia), respectively as monolayer to \up0
\expndtw-3\charscalex100 approximately 80% of confluence in 5% CO\ul0\sub\cf4\f
5\fs21 2\ul0\nosupersub\cf5\f6\fs22 , at 37C. All cells will be purchased from Am
erican Type Cell \up0 \expndtw-1\charscalex100 Collection (USA). Cells will be t
reated with aloe emodin at IC\ul0\sub\cf4\f5\fs21 50\ul0\nosupersub\cf5\f6\fs22
value for 48 hours. Total RNA from cells \up0 \expndtw-3\charscalex100 (MCF-7-a
nd MCF-10A) will be isolated with the RNeasy Mini Kit (Qiagen, Stanford, CA). Al
l samples will \up0 \expndtw-3\charscalex100 be processed following the manufact
urer\u8217?s directions. For protein extraction, the same group of samples will
\up0 \expndtw-3\charscalex100 be harvested and disrupted following the standard
protocol of protein isolation. \par\pard\ql \li1134\sb194\sl-287\slmult0 \up0 \e
xpndtw-2\charscalex100 \ul0\nosupersub\cf9\f10\fs28 3.2.\ul0\nosupersub\cf10\f11
\fs28 \ul0\nosupersub\cf6\f7\fs24 Phase II \par\pard\qj \li1134\ri951\sb0\sl-2
80\slmult0\fi357 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf5\f6\fs22 The e
xtracted mRNA will be converted to biotin-labeled complementary RNA using The Me
ssageAmp \line \up0 \expndtw-2\charscalex100 aRNA kit (Ambion Inc, USA). It
then will be hybridized to HG-U133A Affymetrix oligonucleotide \line \u
p0 \expndtw0\charscalex100 microarrays (Santa Clara, CA). The arrays will be was
hed and scanned according to standard Affyrmetrix \line \up0 \expndtw0\charscale
x100 protocol (www.affymetrix.com). From the microarray results, about 20 gene
s will be chosen for further \line \up0 \expndtw-1\charscalex100 validation. A r
eal time RT-PCR will be carried out to justify the microarray analysis. For prot
ein study, the \line \up0 \expndtw-2\charscalex100 first-dimensional isoelectic
focusing \up0 \expndtw-2\charscalex100 (IEF) will be performed followed b
y second-dimensional SDS-\par\pard\qj \li1134\ri950\sb0\sl-280\slmult0 \up0 \e
xpndtw-2\charscalex100 polyacrylamide gel electrophoresis (SDS-PAGE). After elec
trophoresis, in-gel proteolytic digestion will be \line \up0 \expndtw-2\charscal
ex100 carried out following mass analysis using the MALDI-Tof MS. Spots of the M

ALDI ProteinChip array with \line \up0 \expndtw-2\charscalex100 hydrophobic surf

ace will be pretreated with 2uL of 50% acetonitrile containing 0.5% of trifluoro
acetic acid \line \up0 \expndtw0\charscalex102 (TFA).
Protein
will
be
identified
by
searching
in
publicly
available
NCBI
databa
se \line \up0 \expndtw-3\charscalex100 (www.proteomicmetrics.com) using the pept
ide mass fingerprint generated by trypsin digestion. \par\pard\ql \li1134\sb195\
sl-287\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf9\f10\fs28 3.3.\u
l0\nosupersub\cf10\f11\fs28 \ul0\nosupersub\cf6\f7\fs24 Phase III \par\pard\qj
\li1133\ri952\sb0\sl-280\slmult0\fi357 \up0 \expndtw-2\charscalex100 \ul0\nosup
ersub\cf5\f6\fs22 Microarray data will be analyzed using GeneSpring GX s
oftware.
Protein identification will be \up0 \expndtw-3\charscalex100 dete
rmined using mass spectrometry and database searching. \par\pard\ql \li1134\sb16
5\sl-322\slmult0 \up0 \expndtw-6\charscalex100 \ul0\nosupersub\cf9\f10\fs28 4.\u
l0\nosupersub\cf10\f11\fs28 \ul0\nosupersub\cf9\f10\fs28 Conclusion \par\pard\
qj \li1134\ri951\sb93\sl-280\slmult0\fi357 \up0 \expndtw0\charscalex104 \ul0\nos
upersub\cf5\f6\fs22 This paper provides extensive literature on Ae as anticancer
agent besides giving an insight of the \up0 \expndtw-2\charscalex100 research f
ramework to be carried out. There are two ways in assessing the gene expression
profile either by \up0 \expndtw0\charscalex104 measuring the final product which
is protein or its intermediate, the mRNA. DNA or oligonucleotides \up0 \expndtw
-1\charscalex100 microarrays have been used widely for the analysis of transcrip
tion study; however expression in mRNA is \up0 \expndtw-1\charscalex100 not nece
ssarily reflected to the changes at protein level. Monitoring on the mRNA level
is not sufficient to \up0 \expndtw-2\charscalex100 provide significant informati
on on the various modifications occurred during post-translational stage, which
\up0 \expndtw-2\charscalex100 is crucial for protein functions. Therefore, this
study was designed to tackle the challenge whereby gene and \up0 \expndtw-1\char
scalex100 protein profiling will be carried out concurrently. The findings of th
e expected results from this experiment \up0 \expndtw-3\charscalex100 may provi
de indications of the differential expression of gene and protein that
may reveal underlying \up0 \expndtw0\charscalex102 mechanism of Ae in breast
cancer cell lines thus suggesting the Ae as a promising candidate for natural \u
p0 \expndtw-4\charscalex100 chemoprevention agent in breast cancer treatment. \p
ar\pard\ql \li1134\sb186\sl-322\slmult0 \up0 \expndtw-6\charscalex100 \ul0\nosup
ersub\cf9\f10\fs28 5.\ul0\nosupersub\cf10\f11\fs28 \ul0\nosupersub\cf9\f10\fs28
Acknowledgements \par\pard\ql \li5862\sb0\sl-207\slmult0 \par\pard\ql\li5862\s
b0\sl-207\slmult0 \par\pard\ql\li5862\sb0\sl-207\slmult0 \par\pard\ql\li5862\sb1
52\sl-207\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf14\f15\fs18 43
\par\pard\sect\sectd\fs24\paperw11900\paperh16840{\shp
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0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a

0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
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0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
0028a28a00ffd9}}}}}\pard\sb0\sl-240{\bkmkstart Pg4}{\bkmkend Pg4}\par\pard\li113
3\sb0\sl-253\slmult0\par\pard\li1133\sb0\sl-253\slmult0\par\pard\li1133\sb0\sl-2
53\slmult0\par\pard\li1133\sb170\sl-253\slmult0\fi357\tx8399\tx9358 \up0 \expndt
w-1\charscalex100 \ul0\nosupersub\cf5\f6\fs22 This research is funded by
Exploratory Research Grant Scheme\tab \up0 \expndtw-2\charscalex100 (ER
GS),\tab \up0 \expndtw-2\charscalex100 (Project Code:\par\pard\li1133\sb26\sl253\slmult0\fi0 \up0 \expndtw-1\charscalex100 ERGS/1/2011/SKK/UITM/02/15) and Fu
ndamental Research Grant Scheme (FRGS), (Project Code: 600-\par\pard\li1133\sb28
\sl-253\slmult0\fi0 \up0 \expndtw-1\charscalex100 RMI/ST/FRGS 5/3/Fst(73/2010) g
rants from the Ministry of Higher Education (MOHE), Malaysia.\par\pard\li1133\sb
177\sl-322\slmult0\fi0\tx1491 \up0 \expndtw-3\charscalex100 \ul0\nosupersub\cf9\
f10\fs28 6.\tab \up0 \expndtw-3\charscalex100 References\par\pard\li1133\sb147\s
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]\ul0\nosupersub\cf20\f21\fs20 \ul0\nosupersub\cf7\f8\fs20 D. Jasso de Rodr\u30
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23\fs20 Industrial Crops and Products.\ul0\nosupersub\cf7\f8\fs20 2005, \ul0\no
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-3\charscalex100 \ul0\nosupersub\cf7\f8\fs20 (1): 81-87. \par\pard\qj \li1134\ri
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unds in diabetic \line\tab \up0 \expndtw-4\charscalex100 rats\ul0\nosupersub\cf2
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0\nosupersub\cf21\f22\fs20 59\ul0\nosupersub\cf7\f8\fs20 (3): 195-201. \par\par
d\ql \li1134\ri1237\sb61\sl-280\slmult0\tx1559\tx1559 \up0 \expndtw-3\charscalex
100 [18]\ul0\nosupersub\cf20\f21\fs20 \ul0\nosupersub\cf7\f8\fs20 J. Somboonwon
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ion of Thailand = Chotmaihet Thangphaet.\ul0\nosupersub\cf7\f8\fs20 2000, \ul0\
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\li5862\sb61\sl-207\slmult0 \up0 \expndtw-2\charscalex100 \ul0\nosupersub\cf14\
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0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a0028a28a
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