1

Atsuo Yanagisawa, M.D., Ph.D. Japanese College of IV Therapy

Effect of Vitamin C and anti-oxidative nutrition on radiation-

induced gene expression in Fukushima nuclear plant workers

!"#$%

!&#$%

There are many scientific studies demonstrate protective effects of Vitamin C upon radiation-induced cellular injury .

!"#$%&'()*+,　'%()*+,-.%/01,.%/+-(2%-**34,(2%0**)..,4%3(%5666%3(%7-/-(8%9-:,(2;%1<0%1,.,%,=/0;,4%20%<3><%40;,;%0?%.-43-:0(%4,@,+0/,4%;,@,.,%A0(,%B-..01%?-3+).,C%-(4%2<,.,-D,.%)(4,.1,(2%;2,B%*,++%2.-(;/+-(2-:0(8%E01,@,.C%2<,F%4,@,+0/,4%;,@,.,%GH%4-B->,%132<%43-..<,-%-(4%A+,,43(>C%-(4%,@,(2)-++F%43,4%?.0B%B)+:/+,%0.>-(%?-3+).,8%

-'./'.#/0.*/123/4156$('73$15$3+1-'.8.*/619./4#:1;#6/'(3*/.6<*#:1=>*+'(0.13*1?3$.1@.$.383*&1#1?#6638.150()*/1(A1@#+3#<(*B1

!"#$%%&'(#)'*%+,-,).,%/,012'*%3#**,4,%5%%6"#7)0%8,*-9%+,-,).,%!#"2,%1)%:';')%%

<'$'$#=#%,=%'*>%:%?'01'=%?,.%CDED@ABCBDA9BA>%

I5J%K32-B3(%L%3;%-%/.02,*20.%->-3(;2%,=2,.(-+%.-43-:0(

T. Yamamoto et al.148

al after 14 Gy WBI. We preliminarily studied dose response curve of treatment with ascorbic acid in the irradiated mice. The mice were pretreated with 1.5, 15, 150, and 1,500 mg/kg/day of ascorbic acid for 3 days before 14 Gy WBI. They then received BMT 24 h after radiation. Pretreatment with

150 mg/kg/day of ascorbic acid rescued some subjected mice, although pretreatment with other doses of ascorbic acid rescued no mice (data not shown), thereby studying 150 mg/kg/day of ascorbic acid in the following experiments. Oral intake of 1,500 mg/kg/day ascorbic acid induced potent

Fig. 1. Mouse survival after whole body radiation (A). Mice were irradiated at 6, 8, 10, 12, and 14 Gy. The effect of bone marrow transplantation on mouse survival after radiation (B). Mice were irradiated at 10, 12, and 14 Gy. Mice received BMT 24 h after radiation. N = 12 in each group. *P < 0.01 vs. other groups.

Fig. 2. Radiation-induced damage to the bone marrow and small intestine in mice 7 days after radiation. The mice were irradiated at 0, 8, and 14 Gy. Seven days after whole body radiation, they were euthanized to remove the bone marrow and small intestine, and radiation-induced damage to these organs was examined. Arrows indicate severe radiation-induced degeneration of the small intestine (C-b). Images shown are representative of each mouse group (n = 5).

M0);,%*0)+4%;).@3@,%+,;;%2<-(%2<-(%&%GF%1<0+,%A04F%3..-43-:0(%NOPHQ8%P0(,%B-..01%2.-(;/+-(2-:0(%NPMRQ%"S%<%-D,.%3..-43-:0(%.,;*),4%B3*,%.,*,3@3(>%+,;;%2<-(%5"%GF8%E01,@,.C%(0%B3*,%.,*,3@3(>%5S%GFTOPH%;).@3@,4C%A,*-);,%0?%.-43-:0(T3(4)*,4%GH%;F(4.0B,8

U-B-B020%,2%-+8%7%V-43-2%V,;%"#5#WX5J5SXT5XY8

Prevention of Radiation GI Syndrome by Ascorbic Acid 149

diuretic effect in mice, leading to severe dehydration and thereby exacerbating mortality. Next, we examined the effect of pretreatment or post-treatment with ascorbic acid on the irradiated mice. Mice were treated with 150 mg/kg/day of ascorbic acid for 3 days before or after 14 Gy WBI and they also received BMT at 24 h after radiation. Pretreatment with ascorbic acid before 14 Gy WBI dramatically improved mouse survival (42% survival), whereas treatment with

ascorbic acid after radiation did not affect survival (no mice survived) (Fig. 3). When the mice did not receive BMT fol-lowing 14 Gy WBI, pretreatment with ascorbic acid rescued no subjected mice (Fig. 3). The rescued mice that were pre-treated with ascorbic acid and received BMT after 14 Gy WBI were healthy 30 days after radiation, although they showed marked radiation-induced depigmentation in the fur (data not shown).

Fig. 3. The effect of treatment with ascorbic acid on mouse survival after radiation. The mice were irradi-ated at 14 Gy and received BMT 24 h after radiation. They received ascorbic acid p.o. for 3 days either before or after radiation, or received distilled water only before radiation. The mice pretreated with ascorbic acid were also irradiated at 14 Gy without BMT. N = 12 in each group, *P < 0.01 vs other groups.

Fig. 4. The effect of pretreatment with ascorbic acid or BMT following WBI on the bone marrow or small intestine in the irradiated mice. The mice were pretreated with ascorbic acid p.o. or distilled water for 3 days and then irradiated at 14 Gy. One day after radiation, they received BMT or did not. The mice were euthanized 7 days after radiation to assess radiation-induced damage to the bone marrow and small intestine. Data shown are representative of each mouse group (n = 5).

K32-B3(%L%FEGD10&H%&H+#>I11-;%0.-++F%-4B3(3;2,.,4%?0.%Z%4-F;C%-(4%2<,(%2<,%B3*,%)(4,.1,(2%5S%GF%3..-43-:0(8%9.,2.,-2B,(2%132<%-;*0.A3*%-*34%B-.[,4+F%3B/.0@,4%.-43-:0(T3(4)*,4%3(2,;:(-+%4-B->,C%2<,.,AF%.,;*)3(>%B3*,%.,*,3@3(>%OPH%-2%5S%GF%3(%*0BA3(-:0(%132<%PMR8

12

J*1?#'$41EKL1CDEEL1/4.1'.6$).1/.#01(A1M#<(*#:1=.:ANO.A.*6.1P('$.1/((%1QJR5?JM1S124.*1/4.>12.'.113*1/4.1P)%)6430#1M)$:.#'1-:#*/B1

\O<,(%1,%)(4,.2-[,%2<,%.,;*),%0?%@3*:B;%?.0B%-%.-43-:0(T*0(2-B3(-2,4%-.,-%]);2%-D,.%-%.-43-:0(%-**34,(2%0.%2,..0.3;BC%32%3;%3B/0.2-(2%?0.%.,;*),%2,-B%B,BA,.;%20%T'(0T/:>1/#%.1Q3/#03*1S10.-++F8̂ %

5)/4('61(A1M#<(*#:1O.A.*6.1?.+3$#:1S(::.&.1U1;'()*+11=.:AN1O.A.*6.1P('$.13*1V#T#*1$(00.*/.+13*1/4361#'<$:.L1　!

H(2.-T2,;:*)+-.%3(],*:0(%0?%H043(,T5Z5%4,*.,-;,4%;/,.B%<,-4%;).@3@-+%3(%B3*,%%-(4%3;%[(01(%-;%-(%,=/,.3B,(2-+%B04,+%0?%3(2,.(-+%.-43-:0(%3(]).F8%

9.,2.,-2B,(2%0?%3(2.-T2,;:*)+-.%3(],*:0(%0?%K32-B3(%L%%0.%0.-+%3(2-[,%0?%K32-B3(%L%3(*.,-;,4%;/,.B%<,-4%;).@3@-+%-D,.%2<,%3(],*:0(%0?%H043(,T5Z5%3(%B3*,8%

R<);C%@32-B3(%L%3;%-%.-430/.02,*20.%1<,(%.-430()*+34,;%-.,%3(*0./0.-2,4%3(%2<,%A04F%-(4%4,+3@,.%2<,%40;,%3(%-%*<.0(3*%?-;<30(8

Q3/#03*1S1#61#1@#+3(T'(/.$/('1111111115&#3*6/1J(+3*.NEKE1J*1Q38(1

M#''#1Q@1./1#:B1V1M)$J?.+1EWWKXKYZ[K\N[YD

I"J%K32-B3(%L%3;%-%/.02,*20.%->-3(;2%3(2,.(-+%.-43-:0(

P+004%;-B/+,;%1,.,%2-[,(%A,?0.,%-(4%5%<%-D,.%A.,-[?-;2%132<%0.-+%3(>,;:0(%0?%@32-B3(%L%NZX%B>_[>Q8%%

`-B/+,;%1,.,%3BB,43-2,+F%3..-43-2,4%);3(>%-%L0A-+2TY#%<02;/02%-(4%ab'%4-B->,%1-;%B,-;).,4%);3(>%2<,%c*0B,2c%-;;-F8%%

K32-B3(%L%3(>,;:0(%;<01,4%-%;3>(3d*-(2%.,4)*:0(%3(%ab'%4-B->,%4),%20%3..-43-:0(%-(4%32;%/,-[%,e,*2%1-;%S%<0).%-D,.%3(2-[,%0?%@32-B3(%L8%%

]^.$/1(A1+3./1#*+183/#03*1S1(*1OM516/'#*+17'.#%#&.13*1A'.64:>N36(:#/.+14)0#*1243/.17:((+1$.::6B1

;'..*1?_91./1#:B1?)/#<(*1@.6.#'$41KE[FEWWYIWENEDC

IZJ%f.-+%3(2-[,%0?%@32-B3(%L%/.,@,(2%.-43-:0(T%%%%%%%%3(4)*,4%ab'%4-B->,%3(%<)B-(;8%

•  f.-+%3(2-[,%0?%-+/<-T+3/03*%-*34%-(4%@32-B3(%g%.,4)*,4%).3(-.F%.-430-*:@32F%-(4%0=34-:@,%;2.,;;%3(%3..-43-2,4%*<3+4.,(%3(%L<,.(0AF+8%

•  R<,.,%-.,%()B,.0);%;*3,(:d*%.,/0.2;%-A0)2%()2.3:0(%-;%-%.-430T/.02,*20.%;)*<%-;%K32-B3(%'C%LC%gC%hC%%`,+,(3)BC%G+)2-2<30(,C%M,+-20(3(C%/.0A30:*;C%V,;@,.-2.0+C%,2*8%

•  '(:T0=34-:@,%.,;,.@,%0?%2<,%A04F%3;%-%[,F%.0+,%?0.%/.,@,(:(>%.-43-:0(%3(]).F8%

V-43-:0(%3(]).F%*-(%A,%/.02,*2,4%%AF%-(:T0=34-:@,%()2.3:0(;%%

`('%3*&1;'()T1(A1R4.1=/#/.0.*/15/6)(1a#*#&36#2#1?BOB 1 1?#6#6431b2#7)L1?BOB1")'/(*1]B1")'%6(*L1?BOB11 1"'#+A('+1=B1`..%6L1?BOB11@(*#:+1_)**3*&4#%.L1?BOB1 1=/.8.*1_3$%.>L1-4BOB1R4(0#619.8>L1?BOB1

It is our strongest recommendation that those living in the effected areas regularly take antioxidant supplements such as vitamin C to counteract the negative consequences of long-term low dose radiation exposure as well as to protect the health of coming generations.

fi*3-+%`2-2,2B,(2%0?%7LHR%-A0)2%%\g(@3.0(B,(2-+%.-430-*:@32F%-(4%<,-+2<^%%

%3;;),4%3(%M-.*<%"6C%"#558%

a.8%V0(%E)((3(><-[,%%%%%%　　　 　　　a.8%`2,@,%E3*[,F　　　　　a.8%'2;)0%U-(->3;-1-

a.8%R<0B-;%g8%j,@F%%%%%%%a.8%P).20(%M8%P,.[;0(%%%a.8%P.-4?0.4%`8%O,,[;%%a.8M-;-;<3%k1-A)

-.(T:.124(14#8.1#1T(6637:.13*/.'*#:1#*+H('1.c/.'*#:1'#+3#<(*1.cT(6)'.164():+1/#%.1#*<N(c3+#<8.16)TT:.0.*/61/(10#3*/#3*1(T<0#:1#*<N(c3+#<8.1'.6.'8.1(*1/4.3'1(2*17(+>B1

%%K32-B3(%L% % % % %%%%5TZ>%% % %ZTS%:B,;%-%4-F%%%0.%j3/0;0B-+%K32-B3(%L%5T"%>% % % %213*,%-%4-F%%%'+/<-T+3/03*%-*34 % %%%%5##TZ##%B> %213*,%-%4-F%%%`,+,(3)B% % % % %%%%%%X#T"##l> % %213*,%-%4-F%%%K32-B3(%g % % % %%%%5##T"##%B>% %213*,%-%4-F%

%%132<%02<,.%,;;,(:-+%@32-B3(;%-(4%B3(,.-+;8%

7LHR%T%V,*0BB,(4-:0(%

N5Q%K32-B3(%L%"X>%38@8%A,?0.,%10.[3(>%-2%()*+,-.%/+-(28%%%%%%%.-43-:0(T*0(2-B3(-2,4%-.,-8%%%%%%%%%%`2,.3+,%1-2,.%%%% % %"X#Bj%% % %X#$%@32-B3(%L%%% %X#Bj%%%%%%%%%"#$%M>T*<+0.34,% %%%%%%XBj %%%%%%%%%%%5$%P5"% % %%%5Bj%%%%%%%%%PT*0B/+,=%5##%%% %%%%%%%%%%%%%%5Bj %%%%%%%%%5#$%PY%%% % %%%5Bj%%%%%%%%%"X$a,=/-(2<,(0+ %%%%%%5Bj%

N"Q%a-3+F%;)//+,B,(2;%-2%10.[%%%j3/0;0B-+%K32-B3(%L% %%"%>%m % %2<.,,%:B,;%-%4-F%%%'+/<-T+3/03*%-*34% %Z##%B>%m % %213*,%-%4-F%%%`,+,(3)B% % % %"##%l>%%m % %213*,%-%4-F%%%K32-B3(%g% % % %"##%B>%m %% %213*,%-%4-F%

%%132<%02<,.%,;;,(:-+%@32-B3(;8%

7LHR%.,*0BB,(4-:0(%?0.%/,0/+,%1<0%10.[;%-2%.-43-:0(T*0(2-B3(-2,4%-.,-%3(%2<,%n)[3;<3B-%%b)*+,-.%9+-(28%

b-B,+,;;%n-*,+,;;%%E,.0,;

Japanese College of Intravenous Therapy

(JCIT)

　We are 400 members: 379 physicians 12 dentists 9 veterinarians

　More than 10 educational seminars every year. IV Vitamin C, IV Lipoic acid, IV Chelation,

Myers’ Cocktail, Ukrain, Nutrition therapy, etc.

24

"#!

The 1st JCIT Symposium of ” IVC for Cancer Patients” Oct 17, 2010, Tokyo, Japan

26

$%&'!()*+!,-.!/012!3'34'&!567+758!%9'&!&*:7%;<&%('5=>'!3':75*6!5%+8?6(*=%+!(%!!

()'!/*<*+'8'!57=@'+!*6%+A!!()'!/012!8(*('3'+(B!!!

27

Effect of Vitamin C and anti-oxidative nutrition on radiation-induced gene expression in Fukushima nuclear plant workers - A pilot study

Atsuo YANAGISAWA, M.D., Ph.D. (1) Michie IWATA, M.D., Ph.D. (2) Shuichi AKIYAMA (3)

(1) Japanese College of IV Therapy

(2) Iwata Ladies Clinic,

(3) Gene Science Co, Ltd.

H(2.04)*:0(%

In the statement of “Environmental Radioactivity and Health” issued in March 29, 2011, we strongly recommended that “People living in the effected areas should regularly take antioxidant supplements such as vitamin C to counteract the negative consequences of long-term low dose radiation exposure as well as to protect the health of coming generations.” However, TEPCO, government and all medias ignored our statement.

Recently, we examined cancer-related gene expression in 16 men who had worked at severe radiation environment at Fukushima nuclear plant. We also confirmed the improvement of cancer risk score by the supplementation of Vitamin C and other anti-oxidative nutrition.

28

M,2<04;%N5Q%%`2)4F%;)A],*2;%

`)A],*2;J%5Y%B,(%NZ"TX6%F.;8Q%1<0%10.[,4%XTY%1,,[;%-2%.-43-:0(%*0(2-B3(-2,4%-.,-%0?%n)[);<3B-%b)*+,-.%9+-(2%-D,.%M-.*<%5"C%"#558%'++%;)A],*2;%1,.,%2,B/0.-.F%10.[,.%0?%;)AT;)A%*0(2.-*20.%*0B/-(F%0?%Rg9Lf8%

O0.[%0/,.-:0(;%-2%*0(2-B3(-2,4%-.,-J%%%TTo%L0++,*:(>%*0(2-B3(-2,4%1-2,.%%%TTo%M,-;).3(>%.-43-:0(%+,@,+%3(%2<,%/+-(2%-.,-%%%TTo%V,B0@-+%0?%4,A.3;%%%TTo%f/,.-:0(%0?%<,-@F%B-*<3(,.F%

29

M,2<04;%N"Q%%%M,-;).,B,(2%

P+004%;-B/+,%1-;%0A2-3(,4%?,1%4-F;%-D,.%2<,%10.[3(>%/,.3048%H(%S%10.[,.;C%A+004%1-;%0A2-3(,4%-2%A,?0.,%-(4%-D,.%2<,%10.[3(>%/,.3048%%%M,-;).,B,(2J%N5Q%O<0+,%A+004%*0)(2;%-(4%A+004%*<,B3;2.F%N"Q%9+-;B-%+,@,+;%0?%?.,,%ab'%NZQ%Sp%*-(*,.%.,+-2,4%>,(,%,=/.,;;30(%NSQ%L-+*)+-2,%\L-(*,.%V3;[%`*0.,^%

\L-(*,.%V3;[%`*0.,^%1-;%*-+*)+-2,4%);3(>%,q)-:0(%?0.B,4%AF%2<,%+0>3;:*%-(-+F;3;%?.0B%2<,%@-+),%0?%Sp%*-(*,.T.,+-2,4%>,(,%,=/.,;;30(;8%

30

M,2<04;%NZQ%%%%%H(2,.@,(:0(%

31

N5Q  H(2.-@,(0);%K32-B3(%L%"X>%/.,;*.3/:0(%%%`2,.3+,%1-2,.%%%% %"X#Bj%%%%%%%%%%%%%5"8X$%M>`fS %XBj %%%X#$%@32-B3(%L%%% %X#Bj%N"X>Q %%%KTP5J%5"#B> % %KTP"J %%%%%"B>%% %KTPZ%J% %S#B>%%%KTPXJ%%%"XSB>% %KTPYJ% %5"#B>% %KTP5"J%%%%5B>%

N"Q%a-3+F%0.-+%;)//+,B,(2;%%% %jF/0;0B-+%K32-B3(%L%%%5%>% % % %213*,%-%4-F%% %'+/<-T+3/03*%-*34 % %Z##%B>% % %213*,%-%4-F%% %`,+,(3)B% % % % %"##%l>%% % %213*,%-%4-F%%% %K32-B3(%g % % % %"##%B>% %% %213*,%-%4-F%

%M)+:TK32-B3(; % % % % % %213*,%-%4-F%

Fｒｅｅ ＤＮＡ Concentration

Cut-off value <25ng/mL

>25ng/mL　Healthy control 17% (n=400) Cancer patients 38% (n=400)

Free DNA concentration (ng/mL plasma)

0

25

50

75

100

125

150

175

200

32

a3;2.3A):0(%0?%L-(*,.%V3;[%`*0.,%0(%%E,-+2<F%;)A],*2;%-(4%L-(*,.%9-:,(2;8

0

5

10

15

20

25

～10 ～20 ～30 ～40 ～50 ～60 ～70 ～80 ～90 ～100

割合

％

リスク判定値（0～100）

健常者

担がん患者

330*+5'&!C78D!E5%&'!F.;,..G

■5*+5'&!<*='+(8

■)'*6()H!8?4I'5(8

"#!

".!

,#!

,.!

!#!

!.

J

Cancer Risk Score is calculated from the value of 48 cancer-related gene expression using logistic analysis.

34

Plasma Fｒｅｅ ＤＮＡ

ng/mL

9+-;B-%n.,,%ab'%-(4%L-(*,.%V3;[%`*0.,%%3(%n)[);<3B-%O0.[,.;%

KL!

!.

▲

■●

Cancer Risk Score

■

M!"#!

!.

Five workers were treated with IVC twice a month and oral anti-oxidative nutritional supplements. 35

NO.B.#

ng/mL

FｒｅｅＤＮＡ

Cancer Risk Score

Before 2 M Before 2 M

ge,*2%0?%-(:T0=34-:@,%()2.3:0(-+%3(2,.@,(:0(%0(%n.,,%ab'%-(4%L-(*,.%V3;[%`*0.,%3(%n)[);<3B-%O0.[,.;8%%

KL!

!.

"#!

!.

Four workers took once i.v. Vitamin C (25g) therapy at before they went, and continuously took anti-oxidative supplements during the working period. No significant change was seen in both Free DNA and Cancer Risk Score.

36

ng/mLFｒｅｅＤＮＡ Cancer Risk Score

KL!

!.

"#!

!. Before After work Before After work

ge,*2%0?%-(:T0=34-:@,%()2.3:0(%0(%n.,,%ab'%-(4%%L-(*,.%V3;[%`*0.,%A,?0.,%-(4%-D,.%XTY%1,,[;%0?%%

()*+,-.%/+-(2%10.[%-2%n)[);<3B-8%

L0(*+);30(%

37

(1)  We evaluated the effects of radiation exposure on cancer related gene expression in 17 men who worked 5-6 weeks at the radiation contaminated area of Fukushima nuclear plant.

(2)  After working, plasma free DNA was increased in 2 of 13 workers, and cancer risk score was increased in 3. After the 2 months intervention with intravenous Vitamin C and oral anti-oxidative nutritional supplements, free DNA returned to normal level and cancer risk score was significantly decreased (p<0.05).

(3)  No significant change was seen in 4 workers who took once i.v. Vitamin C at before working and continuous intake of anti-oxidative supplements during the working period.

(4)  Increase in cancer risk by radiation exposure can be protected by anti-oxidative nutritional intervention. Fukushima workers in the radiation contaminated area should take anti-oxidative nutrition therapy immediately.

\f(+F%K32-B3(%L%-(4%02<,.%-(:T0=34-:@,%%()2.3:0);%;)//+,B,(2;%*-(%;-@,%2<,%/,0/+,r;%+3?,%?.0B%.-43-:0(%3(]).F8̂ %

T%7LHR%10.[3(>%>.0)/%

54 yrs. female, stomach cancer

3 yrs after IVC (58 yrs) BW= 40 kg

MF%9.0/0;-+%M-3(2-3(3(>%0/:B-+%-(:T0=34-:0(%.,;,.@,%AF%2-[3(>%K32-B3(%L%-(4%02<,.%-(:T0=34-:@,%()2.3:0(%3;%,;;,(:-+%?0.%*-(*,.%/.,@,(:0(%-(4%2.,-2B,(2C%-(2T->3(>C%.-430T/.02,*:0(%-(4%0/:B)B%<,-+2<8%

P"!

Thank you !

http://www. iv-therapy.jp