Results of non specific detoxification in people exposed to agent orange/dioxin

Journal of military pharmaco-medicine n08-2017

RESULTS OF NON-SPECIFIC DETOXIFICATION IN PEOPLE
EXPOSED TO AGENT ORANGE/DIOXIN
Luong Minh Tuan*; Nguyen Hoang Thanh**
Le Ke Son***; Nguyen Dang Dung****
SUMMARY
Objectives: To estimate the result of non-specific detoxification in the people exposed to agent
orange/dioxin. Subjects and methods: The study included 34 people (23 men, 11 women) who live
around the area contaminated with agent orange/dioxin in Danang were found 2.3.7.8-TCDD
in blood in the previous studies. Before and after applying the non-specific treatment using the
Hubbard method principles, full blood samples were taken and analyzed the dioxin
concentration by the gas chromatography high resolusion mass spectrometry in Eurofins
Analysing Center. Cell immune index CD3, CD4, CD8 were analyzed by the Hematology
Department, Military Hospital 103. Humoral immune index IgA, IgG, IgM were analyzed by the
Military Medical - Pharmacertical Reseach Center, Military Medical University. Results: Before
treatment, mean dioxin level in blood of the exposed group was 50.47 ± 130.74 pg/g lipid; after
the treatment, this indicator decreased down to 39.29 ± 99.93 pg/g; the difference was
significant (p < 0.05). The 2.3.7.8-TCDD concentrations and TEQs clearly showed the reduction
trend. Concentrations of IgA, IgG and IgM increased significantly: IgA from 281.37 ± 188.74 to
303.86 ± 185.11; IgG from 1103.8 ± 223.5 to 1259.1 ± 286.01; IgM from 121.29 ± 71.35 to
172.59 ± 70.76. Cell immune index: CD3 from 1341.4 ± 454.64 cell/µL to 172.59 ± 70.76 cell/µL;
CD4 from 766.59 ± 279.43 cell/µL to 969.85 ± 341.93 cell/µL; CD8 from 443.53 ± 194.22 cell/µL to
555.06 ± 178.39 cell/µL. Conclusions: The Hubbard detoxification method can eliminate dioxins
out of the body and improve some immune indexes of people who exposed to agent orange/dioxin./.
* Keywords: Dioxins; Detoxification; Immune index.

INTRODUCTION
From 1961 to 1971, the US Army sprayed
about 80 million litrers of herbicide in the
South of Vietnam (there were 48.6 litres

of agent orange among them - a 50/50
mixture of 2.4-diclorophenoxyacetic acid

and 2.4.5-triclorophenoxyacetic acid, with
contaminated dioxin containing TCDD at
high rate).
Up to now, there is no specific treatment
for the people exposed to agent orange/dioxin.
Major methods are to inhibit and cure the
disorders of the body due to dioxins, including

* Military Instiute of Preventive Medicine
** 103 Military Hospital
*** Ministry of Natural Resources and Environment
**** Vietnam Military Medical University
Corresponding author: Luong Minh Tuan ()
Date received: 10/08/2017
Date accepted: 18/09/2017

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Journal of military pharmaco-medicine n08-2017
strengthening the immune system, antioxidant, protection of liver, natural enzyme
therapy, multi peptide, vitamin therapy and
physiotherapy [6]. All of these treatments
are non-specific, helping to improve the
general constitution for the patients.
No research on detoxification associated
with analysis on dioxins concentration has

been conducted before, during and after
treatment.
Non-specific treatment based on the
Hubbard method principles was created
to promote the process of elimination of
xenobiotic in fat tissue, so that the toxic
pollutants in the body decrease. In fact,
this method can also eliminate other
contaminated compound. It was applied in
many cases over the world and achieved
good effect. However, we carried out this
study with a view to: Evaluating the results
of non-specific detoxification in the people
exposed to agent orange/dioxin.

- Steam bath: combined with taking
vitamins and other nutritions in a designed
process, especially olive oil, mineral and
vitamin PP.
- Nutrition: ordinary regime with some
kinds of fresh vegetable: extra water
consumption to redeem sweating.
In addition, the participants slept enough
as prescription. The period of detoxification
was from 2 to 3 weeks, depending on
personal “finishing situation” (according to
the requirement of the program).
Doing gymnastic in the program plays
an important role. It helps strengthening
blood circulation to thrust out the toxic

chemical which remained in blood. Without
gymnastic, detoxification effect will be poor.
Olive oil, vitamins help to dilute and to
eliminate dioxins from tissues especially
lipid into circulation and excrete them out.
Vitamin PP (niacin) promote excreting
cholesterol so dioxins can be eliminated

SUBJECTS AND METHODS
34 people (23 men, 11 women) living
around Danang air base, which was
contaminated with agent orange/dioxin
have 2.3.7.8-TCDD in their blood according
to the previous studies. Study period was
from September to November, 2012 at
103 Hospital, Ha Noi, Vietnam.

with bile, follow the fecal passage [4].
Before and after applying the non-specific
treatment followed by the Hubbard method
principle, full blood sample were taken
for analysis of dioxins concentration by
the gas chromatography high resolution
mass spectrometry (GC/HR-MS) at Eurofins
Analysing Center (Germany).

Non-specific detoxification contains the
following elements:

Hematology and biochemistry test were

done at 103 Hospital.

- Doing gymnastic running is applied to
strengthen blood circulation, especially
peripheral circulation.

The data was analyzed by a medical
statistic software (Wilcoxon - signed rank
test and t-student test - SPSS16).

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Journal of military pharmaco-medicine n08-2017
RESULTS AND DISCUSSION
Table 1: Distribution of age, sex of study subject.
Age

Subjects of the study
Male (n1 = 23)

Total

Female (n2 = 11)

(n = 34)

Number

Rate (%)

Number

Rate (%)

Number

Rate (%)

20 - 40

10

29.4

3

8.8

13

38.2

41 - 60

10

29.4

7

20.6

17

50.0

61 - 80

3

8.8

1

2.9

4

11.8

X ± SD

47.55 ± 14.33

43.04 ± 12.44

44.5 ± 13.04

p > 0.05

Average age of participants ranged from 44.5 ± 13.04. The age group 41 - 60 took
the highest rate (50%) and from 61 to 80 took the lowest rate (11.8%). The age and
age distribution were not different between male and female groups (p > 0.05).
Some people of the subject group were born after the war in the Southern part of
Vietnam (20 - 40 years old) and still infiltrated by dioxins. Because of their half-life
spent in contaminated areas and exposure with dioxins (about 7 - 11 years), it can be
explained why these people have had dioxins in their body [1, 2, 7, 8]. According to
Nguyen Hoang Thanh (2010), in the study on 26 veterans who took part in the battles
in the Southern part of Vietnam and who now live in uncontaminated sites, nobody has
2.3.7.8-TCDD in their blood although they used to be victims and had 2.3.7.8-TCDD in
their body before. This results were consistent with remarks by many scientists that
2.3.7.8-TCDD in Vietnam had only one source from agent orange sprayed during the
Ranch Hand campaign [2].
Table 2: Results of biochemistry tests before and after detoxification.
Indexes

Subject of the study (n = 34)

p(1-2)

Before (1)

After (2)

4.8 ± 0.84

4.0 ± 0.1.3

> 0.05

Creatinin (µmol/L)

76.33 ± 10.37

82.22 ± 18.33

> 0.05

Protein (g/L)

75.25 ± 5.09

72.23 ± 3.01

> 0.05

Albumin (g/L)

39.18 ± 4.4

41.48 ± 4.27

> 0.05

308.33 ± 76.64

365.65 ± 74.72

> 0.05

Cholesterol (mmol/L)

5.22 ± 1.2

4.56 ± 1.16

< 0.05

Triglycerid (mmol/L)

1.76 ± 1.02

1.72 ± 2.76

> 0.05

ALT (U/L)

31.72 ± 49.96

35.41 ± 30.73

> 0.05

AST (U/L)

27.18 ± 16.81

30.73 ± 15.68

> 0.05

GGT (U/L)

58.33 ± 68.07

54.53 ± 74.14

> 0.05

Ure (mmol/L)

Acid uric (µmol/L)

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Journal of military pharmaco-medicine n08-2017
Before and after applying the Hubbard detoxification method, only cholesterol index
decreased (p < 0.05). Others showed no change. This may be due to the effect of
niacin (its other name is vitamin B3 or vitamin PP), which plays a vital role, present in
more than 200 reactions of the body to release power and delivery matters to build cells.
Niacin produces 2 coenzymes: nicotinamid adenin dinucleotid (NAD) and nicotinamid
adenin dinucleotid phosphate (NADP) with biotin (vitamin H) takes part in analysis of
food to valuable active ingredients for the body. They help to decrease bad cholesterol
(LDL) and increase good cholesterol (HDL).
Table 3: Alteration of hematological indexes after detoxification.
Subject of the study (n = 34)
Indexes

p(1-2)
Before (1)

After (2)

Leukocyte (G/L)

7.76 ± 2.48

8.27 ± 1.87

> 0.05

Erythrocyte (T/L)

4.76 ± 0.69

4.8 ± 0.61

> 0.05

Hemoglobin (g/L)

135.53 ± 0.14.86

133.44 ± 12.39

> 0.05

229.76 ± 50.28

235.09 ± 58.12

> 0.05

Platelet (G/l)

Hematological indexes of study group had no change (p > 0.05). Applying Hubbard
method did not make any changes to blood cell in our study.
Table 4: Alteration of dioxin concentration after detoxification (pg/g lipid).
Study subject (n = 34)
Indexes

p(1-2)
Before (1)

After (2)

0.39 ± 0.14

0.34 ± 0.8

0.019

50.47 ± 130.74

39.29 ± 99.93

0.001

1.2.3.7.8-penta CDD

8.74 ± 7.15

6.90 ± 4.88

0.001

1.2.3.4.7.8-hexa CDD

5.07 ± 3.33

4.14 ± 3.11

0.001

1.2.3.6.7.8-hexa CDD

20.29 ± 14.02

16.42 ± 10.50

0.001

1.2.3.7.8.9-hexa CDD

5.14 ± 3.02

4.02 ± 2.47

0.001

26.21 ± 14.41

23.86 ± 14.33

0.059

304.36 ± 142.16

275.89 ± 136.19

0.73

Lipid (pg/g)
2.3.7.8-tetra CDD

1.2.3.4.6.7.8-hepta CDD
Octa CDD

Before applying the Hubbard detoxification, the mean concentration of 2.3.7.8-TCDD
(pg/g lipid) of exposed group was 50.47 ± 130.74; after the treatment, this index decreased
down to 39.29 ± 99.93. The difference was significant (p < 0.05). Some other congeners
like 1.2.3.4.6.7.8-hepta CDD; octa CDD changed but not significantly (p > 0.05).
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Journal of military pharmaco-medicine n08-2017
Table 5: Alteration of CDFs and TEQ after detoxification (pg/g lipid).

Study subject (n = 34)
Index

p(1-2)
Before (1)

After (2)

2.3.7.8-tetra CDF

0.87 ± 0.64

0.71 ± 0.5

0.011

1.2.3.7.8-penta CDF

1.50 ± 1.13

1.36 ± 0.92

0.0890

2.3.4.7.8-penta CDF

12.68 ± 5.37

10.99 ± 4.90

0.001

1.2.3.4.7.8-hexa CDF

29.46 ± 18.85

26.35 ± 17.31

0.01

12.3.6.7.8-hexa CDF

21.12 ± 15.20

20.35 ± 14.91

0.1

1.2.3.7.8.9-hexa CDF

0.59 ± 0.30

0.49 ± 0.33

0.16

2.3.4.67.8-hexa CDF

1.91 ± 0.75

1.62 ± 0.97

0.35

1.2.3.4.6.7.8-hepta CDF

28.43 ± 16.15

24.45 ± 12.72

0.21

1.2.3.4.7.8.9-hepta CDF

2.24 ± 1.0

1.94 ± 0.97

0.71

Octa CDF

7.26 ± 3.43

5.52 ± 2.25

0.01

WHO (1998)-PCDD/F TEQ excl. LOQ [a]

74.65 ± 134.77

59.65 ± 103.44

0.001

WHO (1998)-PCDD/F TEQ incl. 1/2 LOQ [b]

74.68 ± 134.76

59.69 ± 103.43

0.001

WHO (1998)-PCDD/F TEQ incl. LOQ [c]

74.71 ± 134.75

59.72 ± 103.42

0.001

WHO (2005)-PCDD/F TEQ excl. LOQ [a]

72.11 ± 134.53

57.48 ± 103.17

0.001

WHO (2005)-PCDD/F TEQ incl. 1/2 LOQ [b]

72.14 ± 134.52

57.52 ± 103.16

0.001

WHO (2005)-PCDD/F TEQ incl. LOQ [c]

72.17 ± 134.52

57.57 ± 103.14

0.001

The mean blood concentration of 2.3.7.8-tetra CDF and congeners were 2.3.4.7.8penta CDF; 1.2.3.4.7.8-hexa CDF; octa CDF of the exposed group significantly decreased
after the treatment (p < 0.05).
The decreasing trend in TEQ of dioxins concentration was also seen after detoxification.
Table 6: Results of humoral immune indexex before and after detoxification.
Indexes

Before (1) (X ± SD)

After (2) (X ± SD)

p

IgA (mg/dL)

281.37 ± 188.74

303.86 ± 185.11

< 0.05

IgG (mg/dL)

1103.8 ± 223.5

1259.1 ± 286.01

< 0.01

IgM (mg/dL)

121.29 ± 71.35

172.59 ± 70.76

< 0.05

Concentrations of IgA, IgG and IgM increased significantly after applying the Hubbard
detoxification method (p < 0.05).
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Journal of military pharmaco-medicine n08-2017
Table 7: Results of cell immune indexes before and after detoxification.
Indexes

Before (1) (X ± SD)

After (2) (X ± SD)

p

CD3 (cell/µL)

1341.4 ± 454.64

1575.1 ± 441.08

< 0.01

CD4 (cell/µL)

766.59 ± 279.43

969.85 ± 341.93

< 0.01

CD8 (cell/µL)

443.53 ± 194.22

555.06 ± 178.39

< 0.01

After applying detoxification, the indexes estimated the cell immune corresponding
strain T lympho including CD3, CD4, CD8 both increase significantly with p < 0.05.
Mentioning the immune system of people exposed to dioxin, Phan Thi Phi Phi [2]
and some authors agreed that the immune system was affected and the number of
T cell decreased fast and clearly, which is suitable for our study. After applying
detoxification, the immune indexes increased from low level to the normal level.
Detoxification based on the principles of Hubbard program may improve and harmonize
elements of immune system.
CONCLUSIONS
Before applying the Hubbard detoxification,
the mean blood dioxins concentration of the
exposed group was 50.47 ± 130.74 pg/g
lipid; after treatment this index decreased
down to 39.29 ± 99.93. The difference
was significant (p < 0.05). TEQ also
decreased significantly. Concentrations of
other congeners had no significant changes.
After detoxification, only cholesterol
decreased significantly. Other biochemical
indexes showed no change. Hematological
indexes of study group before and after
treatment showed no difference (p > 0.05).
Concentrations of IgA, IgG and IgM
increased significantly: IgA from 281.37 ±
188.74 to 303.86 ± 185.11; IgG from
1103.8 ± 223.5 to 1259.1 ± 286.01;
IgM from 121.29 ± 71.35 to 172.59 ± 70.76.
Cell immune index: CD3 from 1341.4 ±
454.64 cell/µL to 172.59 ± 70.76 cell/µL;

220

CD4 from 766.59 ± 279.43 to 969.85 ±
341.93 cell/µL; CD8 from 443.53 ± 194.22
cell/µL to 555.06 ± 178.39 cell/µL.
We primarily recognised that non-specific
detoxification based on the principles of
Hubbard program can eliminate dioxins
from the body and improve some immune
indexes of people exposed to agent
orange/dioxin.
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