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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.
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 (luongtuan27@gmail.com) Date received: 10/08/2017 Date accepted: 18/09/2017 215 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 to 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) 216 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 8.8 13 38.2 41 - 60 10 29.4 20.6 17 50.0 61 - 80 8.8 2.9 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 - 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) 217 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 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) 218 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) 219 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 REFERENCES Hue N.T, Nam V.D, Thuong N.V, Huyen N.T, Phuong N.T, Hung N.X, Tuan N.H, Son L.K, Minh N.H Determination of PCDD/Fs in breast milk of women living in the vicinities of Danang agent orange hot spot (Vietnam) and estimation of the infant's daily intake Sci Total Environ 2014 Sep 1;491-492:212-8 Doi10.1016/j.scitotenv.2014.02.054 Epub.2014 Mar 2014 Phan Thi Phi Phi et al The long term effect of AO/dioxin on the immune system blood of AO/dioxin chronic expossed people Vietnam - America Scientific Conference on the Journal of military pharmaco-medicine n08-2017 harms of AO/dioxin on the people and environment Hanoi 2002, pp.162-166 Thanh Nguyen Hoang Project of 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Supratman S, Maruzeni S, Nishijo M, Nakagawa H, Honma S, Nakano T, Takasuga T, Nhu D.D, Hung N.N, Son le K Serum dioxin levels in Vietnamese men more than 40 years after herbicide spraying Environ Sci Technol 2014 Mar 18;48(6):3496-503 doi: 10.1021/es404853h Epub 2014 Mar PubMed PMID: 24552243 2014 Ron Hubbard Clear body clear mind Bridge Publication, Inc 2010 221 ... 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,... Son L.K, Minh N.H Determination of PCDD/Fs in breast milk of women living in the vicinities of Danang agent orange hot spot (Vietnam) and estimation of the infant's daily intake Sci Total Environ... 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