Development of new methods for risk assessment on multi-chemical exposure to human fetus

1.基因表达分析与脐带环境化学品暴露水平评估：为探讨基因表达与脐带暴露水平的关系，选取5对双胞胎进行研究。研究发现，母体作为环境因素之一，影响基因表达。DNA微阵列和实时荧光定量PCR技术检测人脐静脉平滑肌细胞（HUVSMC）暴露于多氯联苯（PCB 77和PCB 126，PCB 153）和3-甲基胆蒽（3 MC，二恶英类化合物）后，eNOS、NPPB和IL-6表达下调。由于这些基因参与血管的构建，可能影响胎儿的发育.鉴于化学物质对DNA表观遗传学的影响，我们采用原位杂交和荧光化学方法对Dmnt、DNA甲基化酶进行了研究.为了解日本人多氯联苯（PCB）暴露现状，对406名日本人（包括婴幼儿）的血液PCB水平进行了分析。母乳喂养的婴儿PCB水平高于非母乳喂养的婴儿。成人PCB水平随年龄增长而增加。多氯联苯作为氯化物的指示物.为探讨宫内化学品暴露与变态反应的关系，我们利用脐带库系统建立了随访系统。

1. Analysis of gene expression and assessment of exposure level of environmental chemicals of umbilical cord: To investigate association of gene expression and exposure level of umbilical cord, five pair s of twin were used. It is found that maternal body, one of the environmental factors, affected gene expression. When human umbilical vein smooth muscle cells (HUVSMC) were exposed to polychlorinated biphenyls (coplanar PCBs: PCB77 and PCB126, non-coplanar PCBs: PCB153) and 3-methylcholanthrene (3MC, one of the dioxin-like compound), eNOS, NPPB, and IL-6 were down regulated by DNA microarray and real-time PCR. Because these genes involved blood vessel construction, it may be affected to fetal development.2. Because it is thought that chemicals affect to epigenetics of DNA, we investigated on Dmnts, DNA methylation enzymes by in situ hybridization and immunohistochemistry.3. To know current situation of PCB exposure of Japanese, we analyzed blood PCB levels of 406 people including infants. PCB levels of infant with breast milk were higher than without breast milk. Adult PCB levels increased as age. PCB was indicator as chlorinated chemicals.4. To investigate association of chemical exposure in utero and allergy, we constructed follow up system using bank system of umbilical cord.