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.
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 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 (phuongk21@gmail.com) Date received: 06/07/2017 Date accepted: 18/08/2017 170 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 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 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 weeks The research was conducted from October, 2014 to April, 2015 Non-specific detoxification process consists of 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 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] 171 Journal of military pharmaco-medicine 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 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 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 < 0.05 173 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 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 Tổng cục Thống kê Báo cáo điều tra lao động việc làm quý năm 2014 2014, tr.4 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 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 Malmberg K.J, Lenkei R, Petersson M et al A short-term dietary supplementation of high doses of vitamin E increases T helper cytokine production in patients with advanced colorectal cancer Clin Cancer Res 2002, (6), pp.1772-1778 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 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 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 Jonathan E Prousky N, MSc Niacin for detoxification: A Little-known therapeutic use Journal of Orthomolecular Medicine 2011, 26 (2) 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.465-473 175 ... 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... Journal of military pharmaco-medicine Changes in blood TNT concentration before and after treatment Table 2: Blood TNT concentration pre- and post -treatment Blood TNT (µg/mL) Before treatment. .. [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