AFB1 Exposure In Pregnant Women In Guangxi Zhuang Autonomous Region And Bioinformatics Analysis Of Human Liver Cells By AFB1

Objective:Extensive exposure to aflatoxin may be one of the reasons for the high incidence of liver cancer in Guangxi Zhuang Autonomous Region.Previous epidemiological data show that liver cancer in Guangxi has regional aggregation.Whether this aggregation is related to the difference in the exposure level and duration of AFB1 has not been seen.Intervention measures for external exposure of AFB1 may not be effective in reducing internal exposure level of AFB1,but to achieve the goal of reducing the incidence of liver cancer,it is effective to reduce the internal exposure level in the population.Therefore,it is of great practical significance to dynamically watch over the AFB1 internal exposure level in Guangxi to reduce the incidence of liver cancer.Based on birth defects cohort study program,This paper takes Guangxi Zhuang people as the research object,and detect internal exposure levels of Aflatoxin B1(AFB1)in pregnant women at different areas,aims at understanding of AFB1 internal exposure level in pregnant women at different areas,so as to formulate rational and effective intervention measures specifically focus on reducing internal exposure of Aflatoxin B1 with reliable scientific evidence.Considering the difference of individual sensitivity to aflatoxin exposure,it is necessary to explore the biological effects of aflatoxin from the molecular level in order tomaking effective intervention,this study also explored the biological effects of AFB1 exposure on the body in order to find early protection measures specifically for pregnant women,Bioinformatics analysis of human liver cell transcriptional spectrum data derived from induced pluripotent stem cells(iPSC)in NCBI GEO public database at non-cytotoxic concentration was mainly conducted,so as to find reliable biomarkers from them and integrate them into the public health detection system for specified pregnant population.Method:1?Epidemiological basic data and serum sample collection: From 2016 to2017,a cohort study project on birth defects in Guangxi Zhuang population was initiated.According to the project design,five Zhuang populations in Debao County,Jingxi City,Long'an County,Pingguo County and Tiandong County,which have a Zhuang population of more than 90%,are selected as the target population,and samples are taken according to strict inclusion criteria.Crowd screening.After the pregnant women signed the informed consent form,they completed the epidemiological baseline(including the mother's basic condition,pregnancy,drinking and smoking,etc.,while ensuring the accuracy of the data,ensuring that the follow-up work can record the characteristic identification information after the newborn is born.,can accurately view their personal data from the hospital system and investigate the pregnancy outcome)investigation and serum sample collection.The information and samples of this study were derived from the information and samples collected from the cohort study project of birth defects in Guangxi Zhuang population.2 ? Experimental study: Human AFB1 albumin(AFB1-alb)levels were determined by double antibody sandwich assay using human AFB1 albumin(AFB1-alb)enzyme-linked immunosorbent assay(ELISA)kit.The antibodywas coated with the purified human AFB1 albumin(AFB1-alb)to prepare a solid phase antibody,and human AFB1 albumin(AFB1-alb)was sequentially added to the coated microwell,followed by HRP labeling.The antibody is detected to bind to form an antibody-antigen-enzyme-labeled antibody complex,and after thorough washing,the substrate TMB is added for color development.TMB is converted to blue under the catalysis of HRP enzyme and converted to the final yellow color by the action of an acid.The color depth is positively correlated with human AFB1 albumin(AFB1-alb)in the sample.The absorbance(OD value)was measured by a microplate reader at a wavelength of450 nm,and the content of human AFB1 albumin(AFB1-alb)in the sample was calculated by a standard curve.3?Bioinformatics analysis: The NCBI GEO public database was searched with AFB1 as the key word to obtain the chip expression matrix.The differential analysis was performed by GO database and KEGG database,and further analyzed by weighted gene co-expression network analysis.Results:1 ? According to the inclusion exclusion criteria,a total of 994 pregnant women questionnaires,birth outcomes and peripheral blood serum samples were collected,including 78 from Debao County,385 from Jingxi City,191 from Longan County,and 170 from Pingguo County.170.The occupation of pregnant women is 486 for farmers,accounting for 48.89% of all occupations.The age of pregnant women is divided into 4 groups,of which the 25-29 age group has the largest number,accounting for 31.39%.There were more people taking folic acid before or during the first trimester,accounting for 87.53%.The number of people drinking during pregnancy was similar to the number of people using cosmetic products,with 45 and 63 respectively.The number of people whosmoked or passively smoked during pregnancy was 72.43%.Pregnant women who regularly exercise regularly account for 40.14%.2?In the collected birth outcome information,there were 522 male infants and 472 female infants,of which 690 were vaginal natural births,accounting for69.42%.Among all babies,there are 58 premature babies,accounting for 5.84%.After the hospital's ARPG score,there were 15 infants with suffocation,accounting for 1.51%.There were 920 infants with normal birth weight,accounting for 92.56%.There were 195 pregnant women in the first pregnancy,accounting for 19.62%;and 355 pregnant women in the first live birth,accounting for 35.71%.There were 48 patients with bleeding volume exceeding500 ml within 24 hours after delivery of the fetus,which was postpartum hemorrhage,accounting for 4.83%.3?The concentration of serum AFB1 albumin adduct(pg/ml)in peripheral blood of pregnant women was detected by enzyme-linked immunosorbent assay(ELISA),with an average of 559.25 pg/ml,a median of 550.44 pg/ml,and a minimum of 356.42 pg/ml.The maximum value is 814.84 pg/ml.Five regions were grouped for analysis of variance and test of homogeneity of variance,P=0.058,indicating homogeneity of variance.By one-way anova,F=1.957,P=0.099,there was no statistically significant difference in AFB1 exposure between the five regions.4?The results of Logistic regression model analysis showed that the factors influencing AFB1 exposure in pregnant women included age(OR=1.210,95%CI=1.061-1.380),smoking OR passive smoking during pregnancy(OR=5.149,95%CI=3.732-7.104).The results also showed that there were no statistically significant differences in occupation,whether to take folic acid before pregnancy or early pregnancy,whether to drink alcohol during pregnancy,whether to use cosmetic products during pregnancy,and whether to take regular exercise during pregnancy(P > 0.05).5 ? Based on peripheral blood serum of pregnant women AFB1 albumin concentrations of the median value as a dividing line,higher than the median value for AFB1 high exposed group,lower than the median value for AFB1 low exposure groups,to the two groups of exposure and birth outcomes,the results showed that pregnant time,production time,postpartum blood loss(ml)and fetal childbirth way,the difference was statistically significant(P < 0.05).There were no significant differences in gender,ARPG score,birth weight(g)and gestational age(P > 0.05).6 ? GSE127791 chip expression matrix was obtained from the GEO database,and three experimental samples and three control samples were selected into the analysis set.Through enrichment analysis of GO database,KEGG database and other databases,differential expression analysis was carried out.Through weighted gene co-expression regulation network analysis and gene enrichment analysis of the obtained results,25 gene modules were identified,and the green,tan and salmon modules with the highest functional similarity were found.Conclusion:1 ? AFB1 exposure was common in pregnant women with no regional differences.The relationship between fetal AFB1 exposure differences and the incidence of liver cancer needs to be further tracked.AFB1 exposure level and age of pregnant women(OR=1.210,95%CI=1.061-1.380)and smoking OR passive smoking during pregnancy(OR=5.149,95%CI=3.732-7.104)are risk factors for pregnant women's AFB1 exposure.In the birth outcome,postpartum blood loss in pregnant women was found to be statistically correlated with thelevel of AFB1 exposure,which studies have shown may increase postpartum hemorrhage in pregnant women.2 ? Short-term in vitro experiments on the toxicity effects of AFB1 exposure showed complex gene regulation patterns,which could provide clues for clinical testing of the toxicity of AFB1 on the body and assessment of the risk of AFB1 exposure,which also indicated that transcriptome could play an important role in rapid screening of carcinogens in vitro.Through bioinformatics analysis,the toxicity of AFB1 is related to the function of small-molecule stress,cell cycle,and p53-related protein,which further explains the pathogenic mechanism of AFB1,provides a theoretical basis for exploring the biomarker of cellular stress level under the exposure of AFB1,and provides bioinformatics evidence for future application.