Study on changing concentration of TNT and some cytokines in workers occupationally exposed to tnt after non-specific detoxification treatment

Journal of military pharmaco-medicine

STUDY ON CHANGING CONCENTRATION OF TNT AND SOME
CYTOKINES IN WORKERS OCCUPATIONALLY EXPOSED TO
TNT AFTER NON-SPECIFIC DETOXIFICATION TREATMENT
Nguyen Kien Cuong*; Nguyen Phuc Thai*; Do Phuong Huong**
Nguyen Xuan Kien***; Nguyen Minh Phuong****
SUMMARY
Subjects and methods: A clinical trial study was conducted on 70 workers in some military
factories with the evidence of trinitrotoluene (TNT) in their blood from October 2014 to April
2015. The non-specific detoxification method had been applied according to the scientific
principle of the Hubbard method for 3 weeks. Results: Mean TNT concentration before
treatment was 0.150 (µg/mL) and reduced sharply down to 0.0154 (µg/mL) after treatment. The
difference was statistically significant (p < 0.05). Mean concentration of erythropoietin (EPO)
pre-treatment was 676.629 (pg/mL) and rose 792.44 (pg/mL) after the treatment (p < 0.05).
Mean erythropoietin receptor (EPOR) before treatment was 4574.23 (pg/mL) and increased up
to 5982.89 (pg/mL) after treatment, the difference was statistically significant (p < 0.05). Mean
concentration of interferon gamma (IFN-γ) was 395.46 (pg/mL) before treatment and reached
621.12 (pg/mL) after the detoxification (p < 0.05).
* Keywords: TNT; Detoxification; Occupational exposure; Women; Erythropoietin; Erythropoietin
receptor; Interferon gamma.

INTRODUCTION
Trinitrotoluene has been widely used in
the defense with the large number of
workers directly exposed to TNT. Until now,
there is no specific detoxification treatment
for TNT. TNT not only damages the blood
cells in bone marrow and periferal [11] but
also inhibites the body's immune system
[4]. Prophylaxis of occupational TNT toxicity

plays an important role in ensuring the
health of workers exposed to TNT.
L.Ron Hubbard has successfully
researched and applied non-specific

detoxification method to a variety of lipidsoluble chemicals such as chlorobenzocyclic hydrocarbons (PCBs…), heavy metals
(lead, mercury), etc... However, this method
has not been conducted for the detoxification
of TNT, one kind of lipid soluble chemicals.
If the nonspecific detoxification for the
TNT workers is effective, how will it affect
some cytokines that plays an important
role in hematopoietic and immune
stimulation system?. From these reasons,
we conducted a study with a view to:
Evaluating TNT, EPO, EPOR and IFN-γ

* Mlitary Institute of Preventive Medicine
** 103 Military Hospital
*** Military Medicine Department
**** Vietnam Military Medical University
Corresponding author: Nguyen Minh Phuong ()
Date received: 06/07/2017
Date accepted: 18/08/2017

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Journal of military pharmaco-medicine
level after treatment with non-specific

detoxification which applied Hubbard's
scientific principles. The results will
contribute to assess therapeutic effects of
non-specific detoxification for TNT exposure
workers.

2. Methods.
- Methodology of clinical trial, evaluation
pre-and post-intervention, with the
permission from the Ethics Board of
103 Military Hospital.
- Assessment of changes in TNT,
EPO, EPOR and IFN-γ levels before and
after non-specific detoxification treatment.

SUBJECTS AND METHODS
1. Subjects.
- 70 workers work for some military
plants, directly exposed to TNT, with the
evidence of TNT in their blood.
- Workers had been selected with no
chronic diseases such as diabetes, high
blood pressure, etc… and no acute diseases
that needs to be cured.
- Volunteer to participate in this research.
- Workers were treated with nonspecific detoxification at Detox Center,
Military Hospital 103 for 3 weeks. The
research was conducted from October,
2014 to April, 2015.

Non-specific detoxification process
consists of 3 following key contents:
Running exercise in 30 minutes before
the dried sauna for 2.5 hours right after
exercise. All the workers had been
supplied niacin and mineral salt, water,
vitamins A, D, C, E, B and vegetable oil
(unsaturated fatty acids).
- Data had been analyzed by SPSS
software. Comparison of mean values of
TNT, EPO, EPOR and IFN-γ before and
after treatment.

RESULTS AND DISCUSSION
1. Characteristics of the subjects.
Table 1: Characteristics of the research subjects.
Subjects (n = 70)
Characteristics
Number

Prevalence %

Male

34

48.6

Female

36

51.4

Average age

42.02 ± 6.26 (min: 25; max: 56)

- The average age of the participants was 42.02 ± 6.26. They are all exposed to
TNT and they have no signs of occupational TNT toxicity. In fact, they can be
considered as healthy people.
- The proportion of female and male workers accounted for 51.4%, and 48.6%,
respectively. The rate of women in our study is basically consistent with the proportion
of women labour structure in the Red River Delta (50.6%) [1].
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Journal of military pharmaco-medicine
2. Changes in blood TNT concentration before and after treatment.
Table 2: Blood TNT concentration pre- and post-treatment.
Blood TNT
(µg/mL)

Before treatment
(n = 70)

After treatment
(n = 70)

Min - max

0.035 - 0.51

0.000 - 0.160

0.150 ± 0.081

0.0154 ± 0.033

p

< 0.05

The plasma TNT level before treatment
was 0.150 (µg/mL), after detoxification
TNT concentration reduced sharply to
0.0154 (µg/mL). The difference was
statistically significant (p < 0.05).

Niacin (vitamin B3) boosts the release of
free fatty acids, which stimulates the
release of accumulated xenobiotics in the
fats and allows them to be excreted
through the skin [6].

The highest TNT level before treatment
was 0.51 µg/mL and after treatment was
0.16 µg/mL.

Reports on TNT dynamics in humans

are still very rare. It was observed
that after 17 days of leaving the
working environment, the presence of
dinitroaminotoluene (one metabolic of
TNT) was still observed. in the urine at
0.06 mg/L (Woolen BH et al, 1986). This
indicated that TNT and its metabolites
eliminated relatively slow in humans [12].
The results of our study revealed that TNT
still accumulated in the body after 3 weeks
of leaving the working environment.

The results showed that the effects of
detoxification for TNT were well-activated.
According to the science principles of
Hubbard methodology, toxins accumulate in
the lipid organization were excreted from
the body by doing exercise, sauna and
mobilizing
lipid metabolism,
which
accelerates TNT clearance more rapidly
than TNT's natural metabolic pathway.

3. Changes in some cytokines before and after treatment.
Table 3: EPO changes before and after the treatment.
EPO (pg/mL)
Min - max

Before treatment

(n = 70)

After treatment
(n = 70)

20.80 - 2566.23

106.73 - 2369.207

676.629 ± 682.27

792.44 ± 564.06

p

Mean EPO levels before and after
treatment were 676.629 pg/mL and
792.44 pg/mL, respectively. The difference
was statistically significant (p < 0.05).
The increase in EPO level after
exercise has been well documented.
172

< 0.05

Which mechanism that regulates and
mobilizes hematopoietic cells after exercise
are still unknown clearly. Kidney is a
major EPO production site, EPO deficiency
is commonly observed in people with

anemia such as renal failure. Experimental


Journal of military pharmaco-medicine
study by Baker et al (2011) showed that in
trained muscle groups, there was an
increase in the expression level of the
EPO up to 93% [2].
In bone marrow, EPO binds to its
receptor (EPOR) which is expressed on
the surface of erythroid progenitors, where
it stimulates the proliferation and suppresses
the apoptosis of newly formed cells.
Under hypoxia, EPO gene transcription is
activated, resulting in an increase in EPO
level in the circulation, afterward EPO
binds to its specific receptor on the
surface of erythroid precursors in the
bone marrow, initiate the JAK2 (Janus
kiansse 2) signal pathway, and increase
the number of red blood cells.
EPO is known to play a role in blood
vessels protection by acting on endothelial
cells and smooth muscle cells in blood

vessels through the mechanism of
inhibiting apoptosis or by stimulating the
production Nitric oxide and vasodilatation
enhancement [10]. From our point of
view, due to the vasodilatation effect that

was made by the combination of EPO and
niacin during the treatment, therefore
enhance the removal of toxins from the
body. It can be said that the enhancement
of EPO after the detoxification provides
positive protective effects.
We suggested that an increasing EPO
after treatment is possibly due to the effects
of sauna, high temperature increasing
vassolidation, leads to increase blood flow
through the skin. Therefore, it can reduce
oxygen-carrying blood flow to organs
especially the kidneys, which activates
the kidneys to boost the production of
EPO by reverse-regulating mechanism.

Table 4: EPOR changes before and after the treatment.
EPOR (pg/mL)
Min - max

Before treatment
(n = 70)

After treatment
(n = 70)

1874.92 - 21483.110

2302.58 - 23680.54

4574.23 ± 2781.41

5982.89 ± 4279.82

p

< 0.05

The average EPOR pre-treatment level was 4574.23 pg/mL, increased to 5982.89 pg/mL
after the treatment, statistically significant differences (p < 0.05).
EPOR is produced by lack of oxygen and leads to an increase in EPO sensitivities.
The interaction between EPO and EPOR creates biological effects. To the best of our
knowledge, the increase in the concentration of EPO and EPO receptors synergizes
and promotes the biological effect of EPO.
Table 5: IFN-γ changes before and after the treatment.
IFN-γ (pg/mL)

Before treatment
(n = 70)

After treatment
(n = 70)

Min - max

4.413 - 2250.775

2.170 - 4394.197

395.46 ± 457.28

621.12 ± 733.02

p

< 0.05

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Journal of military pharmaco-medicine
Mean IFN-γ level after treatment is
higher than that before treatment. Prior to
treatment, the average IFN-γ level was
395.46 pg/mL. After treatment, mean IFN-γ
level rose to 621.12 pg/mL, the difference
was statistically significant (p < 0.05).
The increase in IFN-γ level after nonspecific detoxification may be due to the
use of some vitamins, including the role of
vitamin E and vitamin A during treatment.
Vitamin E has been shown to increase the
CD4/CD8 ratio. Moreover, rising IFN-γ
production has been demonstrated in
patients with colorectal cancer using
vitamin E to enhance immunity [8].
Vitamin E has been known as antiinflammatory and antioxidant abilities, the
use of vitamin E in treatment is quite
widespread and common. Because of the
anti-free radical effect, vitamin E plays a
significant role in treating many diseases

of various organs in the body [9].
In our opinion, the increase in IFN-γ
level after non-specific treatment is not
only the role of vitamins, but also the role
of heat stimulation due to sauna and the
effect of exercise. In experiment, it was
found that hyperthermia increased the
ability of inhibition tumor growth, enhancing
the survivor time of animal with experimental
tumors [5]. After a moderate exercise, M.
Baum (1997) reported an increase in IFN-γ
level among participants, an IFN-γ
concentration of 974 pg/mL (before
practicing) increased to 1,450 pg/mL after
24 hours of doing exercise.
Therefore, from the findings of our study
on changes in IFN-γ levels before and
after non-specific detoxification based on
the Hubbard method, it can be said that
174

non-specific detoxification applied Hubbard
method including combined exercises,
sauna and vitamins supplementation is
effective to stimulate immune system and
enhance the body's resistance.
CONCLUSION
TNT level before treatment was
0.150 µg/mL, and reduced sharply to
0.0154 µg/mL after treatment. The

difference was statistically significant
(p < 0.05).
Mean concentration of EPO pretreatment were 676.629 pg/mL and rosed
up to 792.44 pg/mL after the treatment
(p < 0.05).
Mean EPOR before treatment was
4574.23 pg/mL and increased to 5982.89
pg/mL after treatment, the difference was
statistically significant (p < 0.05).
Mean concentration of IFN-γ were
395.46 pg/mL before treatment and
increased up to 621.12 pg/mL after the
detoxification (p < 0.05).
RECOMMENDATIONS
Step by step to do research on nonspecific TNT detoxification method at
military medicine at baseline in military
factories, workers exposed to TNT should
be periodically treated by non-specific
detoxification to protect and promote their
health.
REFERENCES
1. Tổng cục Thống kê. Báo cáo điều tra lao
động và việc làm quý 2 năm 2014. 2014, tr.4.
2. Baker J.M, De Lisio M, Parise G.
Endurance
exercise
training
promotes
medullary hematopoiesis. Faseb J. 2011, 25
(12), pp.4348-4357.

Journal of military pharmaco-medicine
3. Baum M., Muller-Steinhardt M, Liesen H
et al. Moderate and exhaustive endurance
exercise influences the interferon-gamma
levels in whole-blood culture supernatants.
Eur J Appl Physiol Occup Physiol. 1997, 76
(2), pp.165-169.

8. Malmberg K.J, Lenkei R, Petersson M et
al. A short-term dietary supplementation of
high doses of vitamin E increases T helper 1
cytokine production in patients with advanced
colorectal cancer. Clin Cancer Res. 2002, 8
(6), pp.1772-1778.

4. B runs - Nagel D, Scheffer S et al. Effect
of 2,4,6-trinitrotoluene and its metabolites on
human monocytes. Environmental Science &
Technology.1999, 33 (15), pp 2566-2570.

9. Peh H.Y, Tan W.S, Liao W et al. Vitamin
E therapy beyond cancer: Tocopherol versus
tocotrienol. Pharmacol Ther. 2016, 162,
pp.152-69.

5. Hattori T, Kokura S, Okuda T et al.
Antitumor effect of whole body hyperthermia
with

alpha-galactosylceramide
in
a
subcutaneous tumor model of colon cancer.
Int J Hyperthermia. 2007, 23 (7), pp. 591-598.
6. Jonathan E. Prousky N, MSc. Niacin for
detoxification: A Little-known therapeutic use.
Journal of Orthomolecular Medicine. 2011, 26 (2).
7. Lee F.S, Percy M.J. The HIF pathway
and erythrocytosis. Annu Rev Pathol. 2011, 6,
pp.165-192.

10. Santhanam A.V, d'Uscio L.V, Katusic
Z.S. Cardiovascular effects of erythropoietin
an update. Adv Pharmacol. 2010, 60, pp.257285.
11. U.S Department of Health and Human
Services. Toxicological profile for 2,4,6 Trinitrotoluene. Agency for Toxic Substances
and Disease Registry. 1995.
12. Woollen B.H, Hall M.G, Craig R et al.
trinitrotoluene: Assessment of occupational
absorption during manufacture of explosives.
Br J Ind Med. 1986, 43 (7), pp.465-473.

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