Associations of PNPLA3 and LEP genetic polymorphisms with metabolic-associated fatty liver disease in Thai people living with human immunodeficiency virus

BACKGROUND The prevalence of metabolic-associated fatty liver disease(MAFLD)is a growing public health issue in people living with human immunodeficiency virus(PLWH).However,the pathophysiology of MAFLD is still unknown,and the role of genetic variables is only now becoming evident.AIM To evaluate the associations of gene-polymorphism-related MAFLD in PLWH.METHODS The study employed transient elastography with a controlled attenuation parameter≥248 dB/m to identify MAFLD in patients from a Super Tertiary Hospital in central Thailand.Candidate single-nucleotide polymorphisms(SNPs)were genotyped using TaqMan®MGB probe 5'nuclease assays for seven MAFLD-related genes.Statistical analyses included SNP frequency analysis,Fisher's Exact and Chi-square tests,odds ratio calculations,and multivariable logistic regression.RESULTS The G-allele carriers of PNPLA3(rs738409)exhibited a two-fold rise in MAFLD,increasing by 2.5 times in MAFLD with human immunodeficiency virus infection.The clinical features and genetic patterns imply that LEP rs7799039 A-allele carriers had a nine times(P=0.001)more significant chance of developing aberrant triglyceride among PLWH.CONCLUSION The current study shows an association between PNPLA3 rs738409 and LEP rs7799039 with MAFLD in PLWH.

Antioxidant Activities and Characterization of Polyphenols from Selected Northern Thai Rice Husks: Relation with Seed Attributes

Rice production generates a significant amount of agricultural waste. This study aimed to give results related to the existence of antioxidant phenols in agricultural waste of selected Northern Thai rice varieties. The antioxidant activities, contents of total flavonoids and phenolic compounds in the ethanolic rice husk extract were evaluated. The highest antioxidant activities were found in the variety PES1CMU, with 2,2’-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid and 2,2-diphenyl-1-picrylhydrazyl as 679.66 and 4.16 mmol/(L·g) trolox equivalent, respectively, ferric reducing antioxidant power as 0.87 mmol/(L·g) Fe2+, total phenolic content as 29.90 mmol/(L·g) gallic acid and total flavonoid content as 12.16 mg/g catechin equivalent. Polyphenol compounds were identified mainly by standard polyphenols using the liquid chromatography mass spectrometry, with the highest contents of phytic acid, o-coumaric acid, naringin and kaempferol. The non-glutenous and wetland ecotypes of rice husk samples were the richest in antioxidant activities and polyphenol contents characterized by using principal component analysis. The glutenous rice husk contained higher antioxidant activities than the rest. Interestingly, quercetin is a significant phenolic compound that positively correlated with the overall antioxidant activities of rice husk. This finding will be relevant for future application of rice husk antioxidant components in the production of functional ingredients as well as for the food and pharmaceutical industries.

THAI技术开发厚层稠油油藏井网参数优选

厚层稠油油藏原油黏度高、埋藏深、油藏流体流动性差、动用程度低,采用THAI(从端部到跟部注空气)技术可提高该类油藏的采收率。通过数值模拟对排状布井方式的井网参数进行优选,得出影响因素的敏感性强弱依次为生产井水平段长度、注气量、井距、注气井到生产井水平段端部的距离。在此基础上,进一步分析厚层稠油油藏中不同布井方式的开发效果。结果表明:随着燃烧前缘向前推进,生产井中会产生气窜通道;方案2VIHP的采油速度和采收率优于其他方案;层厚是影响热量传递的重要因素;垂直注气井的开采效果优于水平注气井。

THAI技术及其在稠油开发中的应用

针对常规火烧油层工艺中使用垂直注入井与生产井进行长距离驱替的缺点,开发了一种新的短距离驱油技术:从水平井端部到跟部注空气技术(THAI)。应用THAI技术在驱油过程中可实现就地升温降粘和非混相空气驱,尤其适合于稠油和超稠油的开采。介绍了THAI技术的原理、与其他热采方式相比的主要优势;通过对THAI技术影响因素的分析,给出了实现该技术的关键工艺。

Detection of Purity of Thai Hom Mali Rice by RAPD

[Objective] The aim was to establish more specific, sensitive, accurate and practical method to detect purity of Thai Hom Mali rice. [Method] RAPD method cooperated with two primers of KDML105 and RD15 was established by feeling for the extraction methods of DNA, optimizing concentrations of the factors influencing the results of RAPD such as template DNA, Mg2＋, random primer, dNTPs and Taq polymerase, and screening the random primers. [Result] The optimum RAPD reaction system was 25.0 μl in total volume, containing 4.0-32.0 ng/μl of template DNA, 200.0 μg/L random primer, 2.0 mmol/L Mg2＋, 200.0 μmol/L dNTPs and 1.0 U of Taq enzyme. Then, the Thai Hom Mali rice and non-Thai Hom Mali rice can be distinguished according to the presence or absence of two DNA markers. [Conclusion] The RAPD technology can effectively cover the shortages of identifications by sense and boiling in water; in addition, it is simple, sensitive and low-cost, suitable to be used in routine tests.

食物相暴露下三丁基锡(TBT)在疣荔枝螺(Thais clavigera)体内的吸收和代谢动力学

将牡蛎消化腺分别暴露在1000ng.L-1和100ng.L-1TBT水溶液中4周,然后将染毒的牡蛎消化腺分别投喂疣荔枝螺(Thais clavigera)。经过45d的暴露和30d的净化,我们发现雌雄疣荔枝螺的消化和生殖系统能较快地吸收TBT(吸收速率ku=0.004-0.022.d-1),并且其代谢(生物代谢系数BDI=5.59-23.30)和排出速率(净化速率ke=0.024-0.053.d-1)也相对较快,各器官中TBT的代谢产物MBT占了相对较高的比例,因此TBT在食物链传递过程中没有出现生物放大的现象。此外,TBT有逐渐从雌螺消化系统向生殖系统转移的趋势,并且雌螺生殖系统对TBT的吸收和富集能力(ku=0.006-0.022.d-1,生物放大系数BMF=0.181-0.664)要显著强于雄螺(ku=0.004-0.014.d-1,生物放大系数BMF=0.142-0.376),但其代谢和净化速率(BDI=5.59-10.50,ke=0.024-0.025.d-1)却显著低于雄螺(BDI=11.5-12.4,ke=0.031-0.050.d-1),雌螺的生殖系统被认为是TBT转移和富集的潜在靶器官,这对我们今后开展TBT污染的环境监测和评价具有重要的参考价值。

Rate of anticoagulant use, and factors associated with not prescribing anticoagulant in older Thai adults with non-valvular atrial fibrillation: A multicenter registry

Objectives To investigate the rate of anticoagulant use,the reasons for not prescribing anticoagulant,and the factors associated with non-prescription of anticoagulant in older Thai adults with non-valvular atrial fibrillation.Methods A multicenter registry of patients with non-valvular atrial fibrillation was conducted during 2014 to 2017 in Thailand.Demographic,medical history,antithrombotic medication,non-antithrombotic medication,and laboratory data were collected and analyzed.Data were compared between the older adult (≥ 65 years) and younger adult (< 65 years) groups.The reasons why anticoagulant was not prescribed were collected,and predictive factors were identified.Results A total of 3218 patients (1873 males) with an average age of 67.3 ± 11.3 years were included.Almost two-thirds (61.0%) of patients were in the older adult group.Anticoagulant was prescribed in 2422 patients (75.3%): 81.4% in the older adult group and 65.7% in the younger adult group.The three main reasons for not prescribing anticoagulant were already taking antiplatelets,patient refusal,and bleeding risk.These reasons were more common in older adults as compared to younger adults.Multivariate analysis revealed current use of antiplatelets to be the most important factor that predict the non-prescription of anticoagulant in older population.Conclusions The prevalence of anticoagulant prescription among older Thai adults with atrial fibrillation is 81.4%.Taking antiplatelet drugs was found to be the strongest reason that predicts the non-prescription of anticoagulant in this patient population.A guideline should be developed to optimize the use of anticoagulant and antiplatelet in older adults.

A Thai family with hereditary pancreatitis and increased cancer risk due to a mutation in PRSS1 gene

AIM: To investigate mutation of serine protease 1-cationic trypsinogen (CT, PRSS1) gene in members of a Thai family with hereditary pancreatitis and pancreatic cancer. METHODS: Polymerase chain reaction and direct sequencing were performed to analyze the PRSS1 gene in two members of the family affected by pancreatitis. Allele specific amplification (ASA) method was then developed to detect the mutation of the PRSS1 gene in all available members of the family and normal control subjects. RESULTS: A cytosine (C) to thymine (T) mutation at position 2441 (g.2441C>T) of the PRSS1 gene, which results in a substitution of arginine by cysteine at position 116 (R116C) of CT, was identified by direct sequencing in both clinically affected members of the family but was not found in the unaffected member. This mutation, which might be arising from deamination of methylated cytosine in CpG dinucleotide of codon 116 (CGT>TGT), was also detected by the ASA method in the two affected members and a proband's brother but was not observed in unaffected members and 54 normal control subjects. CONCLUSION: Autosomal dominant pancreatitis with increased cancer risk in the studied Thai family is most likely due to missense (R116C) mutation in the PRSS1 gene.

Characterization of the salivary microbiome in healthy Thai children

Objective: To investigate the composition of the salivary microbiome of 50 healthy Thai children. Methods: A total 76 provinces in Thailand are grouped into 5 geographical clusters based on unique economics, foods and lifestyles. Geographical locations and the results of an oral assessment were also considered. Genomic DNA was extracted from stimulated saliva samples. Subsequently, amplicon libraries were prepared by 16 S Metagenomic Sequencing Library Preparation. The amplicons were sequenced using an Illumina Miseq platform followed by bioinformatics and statistical analyses.Results: The correlation between oral hygiene status and caries history varied from r^2=0.887 to r^2=0.999 in the geographical groups, suggesting oral hygiene status a strong association between caries history. Twenty taxonomic groups were found in all subjects and constituted 93.6%-96.5% of the microbiome. Of these, genus Veillonella and Prevotella showed significant differences in their proportions between the geographical groups(P<0.05). Furthermore, the proportion of Veillonella parvula, as well as Rothia aeria and Rothia dentocariosa tended to increase with worse oral hygiene status, which was also related to higher dental caries history.Conclusions: The differences in the salivary microbiome as related to geographic regions suggest that environmental factors, which may include dietary habits, could influence the predominant bacteria found in the mouth of Thai children, especially the genus Veillonella and Prevotella. The ratio of Veillonella parvula, Rothia aeria and Rothia dentocariosa may be indicators of worse oral hygiene status and future caries in this population.

Heart failure mortality compared between elderly and non-elderly Thai patients

Objective To investigate heart failure mortality compared between elderly and non-elderly Thai patients.Methods This study included patients at least 18 years of age who were admitted to the hospital with a primary diagnosis of heart failure(ICD-10-TM code:150.9)during 2008-2012 according to three major Thailand reimbursement systems(civil servant,social security,and universal coverage systems).Patients were categorized into either the elderly group(age>65 years)or the non-elderly group(age<65 years).Mortality rate and survival analysis were compared between groups.Demographic,underlying disease and comorbid condition data were collected.Cardiovascular and non-cardiovascular death was also analyzed.Results A total of 201,709 patients were included.The average age of patients was 64.9±14.8 years,and the gender proportion breakdown was 84,155(41.7%)males and 117,554(58.3%)females.Just over half of patients(107,325 patients;53.2%)were elderly.Overall mortality rate was 50.8%.The mortality rate at one month,six months,one year,and three years was 11.0%,24.5%,32.5%,and 46.3%,respectively.Elderly patients had a higher rate of mortality compared to non-elderly patients with an adjusted odds ratio(OR)of 1.47(95%CI:1.46-1.49)for all-cause mortality,an OR of 1.25(95%CI:1.23-1.27)for cardiovascular death,and an OR of 1.72(95%CI:1.68-1.75)for non-cardiovascular death(all P<0.001).After adjusting for potential confounders,elderly status remained the second strongest factor associated with increased risk of mortality after heart failure hospitalization following chronic kidney disease.Conclusions The overall mortality rate after heart failure hospitalization was a very high 50.8%.Multivariate analysis revealed elderly status to be an independent predictor of mortality after hospitalization.This finding suggests that improvements are needed related to the quality of care and follow-up given to elderly Thai heart failure patients.

Differences in characteristics of colorectal neoplasm between young and elderly Thais

AIM: To analyze the differences of clinical characteristics of colorectal neoplasm including polyps between the elderly and young Thai patients.METHODS: Colonoscopy database from December 2000 to October 2004 was retrospectively analyzed. There were 1822 eligible patients who underwent colonoscopy (with a mean age of 56.6 years). Patients were classified into two groups: the older age group (aged ≥ 60 years; n = 989) and the younger age group (aged < 60 years; n = 833). Data were recorded on age, colonoscopic indications, tumor location, colonoscopic findings and their related histological findings. RESULTS: Colorectal malignancy related lesions were more often found in the older age group (21%) than in the younger age group (12%). Left-sided lesions were detected more commonly than right-sided in both age groups in approximately two-thirds of all cases. Hematochezia showed greater association with left-sided lesions in the elderly. No relationship was found between age and neoplasm staging and severity.CONCLUSION: The chance of detecting colorectal neoplasm by colonoscopy was higher in the elderly than in the young Thais. However, both groups had the lesions predominantly located in the left side.

Development and characterization of seed gums from Tamarindus indica and Cassia fistula as disintegrating agent for fast disintegrating Thai cordial tablet

Crude seed gum and their carboxymethyl derivatives from Tamarindus indica and Cassia fistula seeds were developed and characterized to apply as the pharmaceutical disintegrant in fast disintegrating Thai cordial tablet. The chemical structure of crude gum was chemically modified via carboxymethylation. Degree of substitution(DS) of carboxymethylated gums was determined. Carboxymethylated gums with minimum and maximum DS values were chosen for further application. IR absorption spectra of gum samples were observed to verify their chemical structure changes. In physical properties, the intrinsic viscosity and swelling property of all gum samples were evaluated. The results showed that carboxymethylated gums had higher intrinsic viscosity than those of crude gum. Moreover, they could swell and be soluble in cold water better than those of crude gums. In conclusion, the modified gums from both plants could provide higher hardness and be better used than that crude gums for fast disintegrating Thai cordial tablet. However, this is a preliminary assessment to expressing pharmaceutical application possibility of these gums as disintegrants, diluents and drug release controlling agents.

New Thai herbal shampoos as pediculicides for killing head louse, Pediculus humanus capitis De Geer(Phthiraptera)

Objective: To evaluate the efficacy of Thai herbal shampoos [Cartoxylum formosum(C.formosum) + eucalyptus essential oil(EO), C. formosum + citrus EO, Solanum trilobatum+ eucalyptus EO, Solanum trilobatum + citrus EO, Moringa oleifera + eucalyptus EO and Moringa oleifera + citrus EO] for killing all stages of Pediculus humanus capitis(Phthiraptera).Methods: A filter paper contact method was applied with three concentrations(0.05, 0.10 and 0.20 mL/cm^2) of each Thai herbal shampoo as well as permethrin pediculicide(positive control) and drinking water(negative control) against eggs, nymphs and adults of Pediculus humanus capitis. Mortality rates of the eggs were recorded after 7 days of incubation while those of nymphs and adults were recorded after 5 minutes of contact. Results: All herbal shampoos at the high concentration were highly effective against nymphs and adults, but not effective against the eggs. C. formosum + eucalyptus EO and C. formosum + citrus EO shampoos at all concentrations exhibited the highest efficacy against nymphs and adults with100% mortality rate at 5 min and LC_(50) values of 0.004 and 0.005 mL/cm^2, respectively. All formulation of Solanum trilobatum and Moringa oleifera shampoos added with eucalyptus EO showed mortality rates against nymphs at 92.0%-100.0% and 76.0%-100.0% and against adults at 84.0%-100.0% and 20.0%-32.0%, respectively. Permethrin pediculicide was not effective against the eggs, but showed 68.0%-92.0% and 28.0%-60.0% mortality rates against nymphs and adults. Conclusions: These results indicate that C. formosum + eucalyptus EO shampoo can be used as an effective nymphicide and adulticide against Pediculus humanus capitis.

Thai men＇s health and sexual attitude

Men＇s health awareness, including the research and study of quality of life, sexual desires and risk factors, has increased worldwide. In Thailand, this advancement is made possible by cooperation, research and sponsorship from the local Thai community. This article aims to illustrate the sexual attitudes of Thai people, to determine the degree of erectile dysfunction （ED） and to investigate how to manage and cope with ED in a Thai community. We reviewed the relevant literature from Thai-based articles and surveys in regard to men＇s health, sexual attitudes, the prevalence of ED and common risk factors in the Thai community. The primary risk factor for ED in Thai men was age-related health decline and the presence of vascular disease. Most Thai men will seek consultation from their partner in regard to ED. The main presentation of metabolic disease in Thai patients was dyslipidemia. New selective serotonin reuptake inhibitors are not available for premature ejaculation in Thai communities. The debate in regard to malpractice compensation is an issue that should be closely monitored. There is currently a shortage of home care for the elderly in Thailand. The insights provided by the articles helped recruit the study patients and in turn, helped us gain knowledge that can be translated into improved men＇s health care in Thailand.

Complete mitochondrial genome of the Thai Red Junglefowl (Gallus gallus) and phylogenetic analysis

DEAR EDITOR,In this study, we sequenced the complete mitochondrial genome （mitogenome） of the Thai Red Junglefowl （RJF; Gallus gallus） using the next-generation sequencing （NGS） platform of the Ion Torrent PGM. Samples were taken from Mae Wang District, Chiang Mai Province, northern Thailand Our data showed the complete mitogenome to be 16 785 bp in length, composed by 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and one control region. The genome nucleotide composition was 30.3% A, 23.7% T, 32.5% C, and 13.5% G, resulting in a high percentage of A＋T （50.4%）. Phylogenetic analysis revealed that the mitogenome belonged to haplogroup X, whereas those of all domestic chickens belong to haplogroups A to G. This newly released mitogenome sequence will advance further evolutionary and population genetics study of the RJF and domestic chicken The availability of the G. gallus mitogenome will also contribute to further conservation genetics research of a unique species, listed as ＇data deficient＇ in Thailand.

Zinc and Copper Status in Urban Thai PregnantWomen and Their Pregnancy Outcome

Zinc and copper intakes and serum concentrations were determined at the 32th and 36th week of pregnancy in 96 healthy Thai women attending a prenatal clinic. Dietary data from 3-day food records revealed that the mean dietary zinc and copper intakes in Thai pregnant women were 6. 9 and 1. 7mg/d (equivalent to 34 and 69% of Thai RDA), respectively. Fifty-five percent of zinc intake was from animal origin, whereas 59% of copper intake was from plant origin.The infants of the 64 women who were in the study until delivery, were all normal.Most (>96%) of the maternal serum zinc and copper concentrations were within normal ranges. Only one woman at 32th week had low serum zinc level (<6. 4μmol/L). There were no significant correlations between the maternal serum zinc or copper levels and pregnancy outcome (birth weight and birth length). In this study, although the zinc and copper intake in most Thai pregnant women (99 and 58%, respectively) were less than two-thirds of the Thai RDAs, they were not at risk of suffering from zinc and copper deficiencies as determined by serum zine and copper concentrations, abnormal parturition and abnormal pregnancy outcome.

Effect of Cooking on the Oxalate Content of Selected Thai Vegetables

Total and soluble oxalate content levels were measured in thirteen selected vegetables purchased from a local market in Surin Province in the northeast of Thailand. Total oxalate contents of the leaves, shoots and roots of the fresh vegetables ranged from 249.5 ± 12.1 to 7597.9 ± 77.6 mg oxalate/100g dry matter (DM) while soluble oxalate content ranged from 205.0 ± 2.3 to 2677.6 ± 19.0 mg oxalate/100 g DM. Very high levels of total oxalates were found in three of the selected Thai vegetables, Polygonum odoratum (7597.9 ± 77.6 mg/100g DM), Piper aurantaucum (7026.6 ± 76.9 mg/100g DM) and Limnophila aromatic (6179.0 ± 23.6 mg/100g DM). However, the soluble oxalate content of L. aromatic was low and the highest soluble oxalate contents of fresh vegetables were found in P. odoratum, P. aurantuacum and Neptunia oleracea at 2677.6 ± 19.0, 2152.2 ± 65.3 and 1640.8 ± 3.4 mg/100g DM, respectively. Boiling the vegetables reduced the soluble oxalate content between 30.4 and 65.0%. The insoluble oxalate levels increased in eleven of the cooked vegetables while small decreases were observed in L. aromatic and N. oleracea.

Radiation effects and radioprotection by Thai medicinal plants in mouse macrophage cell line

Objective:To investigate the effects of radiation on growth-arrested(GA) and micronucleus-production(MP) rates,and the radioprotective properties of Thai medicinal plants in mouse macrophage cell line RAW264.7 in vitro.Methods:Mouse macrophage cell line(RAW264.7) was cultured in vitro.Various radiation exposures, growth-arrested rate assay,micronucleus production assay,and radioprotection by Thai medicinal plants were performed.Results:The results showed that GA and MP rates forγ-rays and UV were dose-dependent. The 50%-affected dose ofγand UV radiation for the GA rate was 10 Gy and 159 microwatt/cm for 0.5 seconds, respectively.After X-ray exposure,there was no apparent effect on RAW264.7 cells,even with a fortyfold human diagnostic dose.Two exposures toγradiation at 20 Gy resulted in a significantly higher MP rate than 20 Gy single exposure or control(P【0.05).The Thai medicinal plants(Kamin-chun capsules,Curcuma longa Linn;Hed lingeu,Ganoderma lucidum;Ya Pakking capsule,Murdannia loriformis) could not prevent cell damage,but epigallocatechin gallate and L-cysteine could provide protection from 2 Gyγ-ray exposure. Conclusion:γradiation caused chromosomal damage during cell division and UV caused cell death, while X-ray radiation was safe.The radioprotective effects of Thai medicinal plants,Kamin-chun,Hed lingeu, and Ya Pakking,could not prevent cell damage in this study.

Trends in the prevalence of obesity among young Thai men and associated factors:From 2009 to 2016

Background:The prevalence of obesity has been increasing in both males and females worldwide.In Thailand,the National Health Examination Surveys reported that the prevalence of obesity(body mass index(BMI)≥30 kg/m^2)among Thai male adults aged 20–59 years increased from 1.7%in 1991 to 6.8%in 2009.Obesity has been confirmed to lead to health problems,including noncommunicable diseases.In the present study,we report trends in the prevalence of obesity among new conscripts from 2009 to 2016.We also investigated the associated factors of obesity.Methods:Serial cross-sectional studies were conducted from 2009 to 2016 among male Royal Thai Army(RTA)conscripts whose weight and height had been measured to determine BMI after being inducted.Each subject completed a detailed risk factor questionnaire.Obesity was defined as BMI≥30 kg/m2.Results:A total of 26,540 young Thai males conscripted into the RTA were included in this study.The prevalence of obesity was 2.2%in 2009,3.4%in 2010,2.5%in 2011,2.9%in 2012,3.4%in 2013,4.4%in 2014,5.0%in 2015,and 4.8%in 2016(P for trend<0.0001).The independent risk factors for obesity were coming from the north central and south regions compared with the northeast,higher education level,indoor occupation and no regular exercise.Conclusions:Our data emphasized that obesity constitutes a serious problem among young Thai men.We could apply these findings in military units to other groups at any age.Regular exercise should be provided to young adults and other age groups to slow the process of obesity,so that associated complications,especially noncommunicable diseases,will cease.

Evolutionary relationship of 5′-untranslated regions among Thai dengue-3 viruses,Bangkok isolates,during 24 year-evolution

Objective:To study evolutionary relationship of the 5'untranslated regions(5'UTRs) in low passage dengue3 viruses(DEN3) isolated from hospitalized children with different clinical manifestations in Bangkok during 24 year-evolution(1977-2000) comparing to the DEN3prototype(H87).Methods:The 5'UTRs of these Thai DEN3 and the H87 prototype were amplified by RT-PCR and sequenced.Their multiple sequence alignments were done by Codon Code Aligner v 4.0.4 software and their RNA secondary structures were predicted by MFOLD software.Replication of five Thai DEN3 candidates comparing to the 1187 prototype were done in human(HepG2) and the mosquito(C6/36) cell lines.Results:Among these Thai DEN3,the completely identical sequences of their first 89 nucleotides,their high-order secondary structure of 5'UTRs and three SNPs including the predominant C90 T,and two minor SNPs including A109 G and A112 G were found.The C90 T of Thai DEN3.Bangkok isolates was shown predominantly before 1977.Five Thai DEN3 candidates with the predominant C90 T were shown to replicate in human(HepG2) and the mosquito(C6/36) cell lines better than the H87 prototype.However,their highly conserved sequences as well as SNPs of the 5'UTR did not appear to correlate with their disease severity in human.Conclusions:Our findings highlighted evolutionary relationship of the completely identical 89 nucleotide sequence,the high-order secondary structure and the predominant C90 T of the 5'UTR of these Thai DEN3 during 24 year-evolution further suggesting to be their genetic markers and magic targets for future research on antiviral therapy as well as vaccine approaches of Thai DEN3.