研究神经管畸形（NTDs）的早期无创诊断方法，使其能够在神经不可逆损伤出现之前获得诊断，对于早期干预有重要意义。本课题将利用网络科学中可以对非线性数据分析与处理的人工神经网络（ANN）技术，将前期研究利用不同组学技术筛选出的21个NTDs 分子标志物（包括miRNA和蛋白质）构建新的NTDs诊断模型，并进行多中心、大样本的验证。同时利用蛋白质组学技术分析NTDs孕鼠血清、胎鼠血清、羊水和胎盘的蛋白质谱变化规律，从母体免疫、应激反应和胎儿本身固有蛋白质两方面综合分析，明确这些分子标志物的来源，同时也可筛选更多新的NTDs特异性分子标志物，不断完善ANN诊断模型。另外，在发现分子标志物中存在大量细胞骨架蛋白这一新现象的基础上，本课题将以表达LMNA和GFP融合蛋白的转基因小鼠为供体，以正常小鼠为受体母鼠进行受精卵移植，系统观察LMNA和GFP融合蛋白进入母鼠血清的时间点和途径，明确其具体机制。

Neural tube defects (NTDs) are the most serious congenital malformation harmful to human health and current therapeutics for NTDs are unsatisfactory. The study on non-invasive early prenatal diagnosis of NTDs can provide clues to guide clinical interventions before the formation of severe structural deformity. Using various high-quality screening technology, we have found 21 new molecule biomarkers (including miRNAs and proteins) in serum of pregnant women with neural tube defects. In the present project, these diversiform biomarkers data will be analyzed in artificial neural network (ANN), a powerful and innovative mathematical method that tracks/categorizes numerical inter-relationships between the nonlinear data, to establish a new prenatal diagnosis model for NTDs. And we will take advantages of the cooperation with a number of domestic birth defects research centers for multicenter and large sample validation. To identify the origin of these molecular markers from the maternal or fetal intrinsic proteins, we will detect the expressive disciplinarian of proteins in the serum from pregnant rats, serum from fetal rats, amniotic fluid and placenta using proteomic technology. At the same time, more novel NTDs specific molecular markers can be discovered, and be used to improve the ANN diagnosis model. Moreover, based on the novel discovery of a large of candidate biomarkers obtained in maternal serum belong to the cytoskeleton family, we will generate a LMNA/GFP transgenic mouse model, and transfer the zygote from LMNA/GFP transgenic mouse to oviducts of a normal mouse recipient. To explore the pathway of cytoskeletal proteins enter into maternal serum, the spatio-temporal expression of LMNA\GFP fusion protein will be detected in the serum of maternal receptor, serum of fetal rats, amniotic fluid, placenta, fetal heart and spinal cord on different development stages. This study not only could promote the application of complex biomarkers in clinical screening of NTDs, and also might offer a further avenue for non-invasive prenatal diagnosis for other congenital malformations disease.