Biomarker Study Of Water-Born Endemic Arsenism In Kuitun, Xinjiang, China

Objective:The goal in this study was to explore the distribution characteristics of water-born endemic arsenism in Kuitun, Xinjiang, chose biomarker in plasma of endemic arsenism patients. To analysis any probable relationship between MT gene, GST gene polymorphisms and endemic arsenism. To contrast the difference expression of proteins from endemic arsenism to normal control.So to discover the mocular mechanism of arsenism and genetic factors on the metabolism of arsenic, to offer specific, sensitive, fastern and economic biomarker for early diagnosis, prevension and risk assessment of Xinjiang endemic arsenism. Methods:This study using cohort study to followed-up 108 individuals who were diagnosed endemic arsenism in Kuitun, Xinjiang in 1985, and matched 95 individuals as intra-control (IC),91 individuals as extra-control (EC) by sex and age. NO and (32-MG of plasma were compared by biochemical tests and radioimmunoassay. We performed genotyping of Metallothionein 2A (MT2A) gene, Glutathione-S-transferase Theta 1 (GSTT1), Glutathione-S-transferase Omega 1 (GSTO1) gene by Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) to investigate the allele frequencies of these polymorphisms in arsenism patients (AP) and IC, EC.Two-dimensional electrophoresis (2-DE) was applied to segregate the total proteins in the plasma form 25 cases arsenism patients and 25 cases health control. then, the differential expression proteins were identified by using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Results:1) AP had no significant difference with IC and EC in hypertension, ECG results abnormal (P> 0.05); in the skin pigmentation, skin hypopigmentation were statistically significant (P<0.05). AP and IC had statistically difference in skin hyperkeratosis (P<0.05), had no statistically difference with EC. (P> 0.05). After logarithmic transformation and covariance analysis, corrected mean of NO content of blood plasma was statistically significant (P<0.05) for AP vs. IC, EC controls. Fixed other factors, interaction of groups and drinkingwater time could effect NO content. AP, IC compared with EC,β2-MG of blood plasma was statistically significant (P<0.05), Fixed other factors, interaction of groups and age could effectβ2-MG content, age could also effect; 2) After genotyping GSTT1 and GSTOI (rs4925, rs11509438, rs11509437) polymorphisms, firstly there was no statistically significant difference in AP vs.IC and EC, concerning both, genotype and allele frequencies, respectively (P>0.05). secondly binary logistic regression showed:fixed other factors, the mutation superiority of GSTO1 (rs4925) gene will be increased 0.971, corresponding to drinking-water time increased 1 year (P<0.05); fixed other factors, fale compared to female, the mutation superiority of GSTO1 (rs11509437) gene will be increased 0.439 (P<0.05); 3) The frequencies of MT2A (rs10636) genotypes CC, CG, GG was statistically significant between AP and IC, EC (P<0.05). The frequency of CG>CC>GG, The frequency of CG of EC (68.6%) higher than its of AP (43.6%) (P<0.05). The frequency of GG of AP (2.5%) higher than its of IC (9.9%) and EC (10.5%) (P<0.05).The frequencies of MT2A (rs10636) allele C, G was not statistically significant between AP and IC, EC (P>0.05). Binary logistic regression showed:fixed other factors, gender could effect MT2A genetic polymorphism (P<0.05), the mutation superiority of female higher than male 1.941; 4) To Analysis different expression proteins of 25 cases of blood plasma in Xinjiang endemic arsenism 1:1 matched with normal control. Performed two-dimensional gel electrophoresis and MALDI-TOF-MS proteomic analyses of blood plasma, By the analyses of match differentially displayed map,12 prominent (5up-regulated and 7down-regulated) proteins. Conclusion:Water arsenic content was excessive in Xinjiang Kuitun arsenic exposure area. the health state of inhabitants, as skin lesion.can be used to diagonsis of endemic arsenism. NO of blood plasma was not a biomarker of endemic arsenism. Butβ2-MG of blood plasma was confirmed to be a biomarker of endemic arsenism in this study. The frequencies of MT2A (rs10636) genotypes was statistically significant between AP and IC, EC, so MT2A (rs10636) gene was a biomarker of endemic arsenism.In this study, genotype of GSTT1 and GSTO1 (rs4925, rs 11509438, rs11509437) polymorphisms had no relationship with endemic arsenism, maybe those four gene were not biomarker of endemic arsenism.Performed two-dimensional gel electrophoresis and MALDI-TOF-MS proteomic analyses of blood plasma, By the analyses of match differentially displayed map,12 prominent (5up-regulated and 7down-regulated) proteins, each being unique in endemic arsenism were identified with mass spectrometry and bioinformatic database queries. These identified proteins were immune response protein, neural degeneration protein, male reproduction protein, antioxidant related proteins, those protein could be biomaker of endemic arsenism and might be for the diagnosis, treatment and prevention of endemic arsenism in the future.