Pregnant women are more vulnerable to iodine deficiency and iodine excess. The study aimed to assess the changes in iodine nutrition and thyroid function of pregnant women exposed to different iodine sources resulting in various iodine intakes during pregnancy. From 2016 to 2017, 2004 healthy pregnant women aged 20-35years from Shandong and Tianjin, China, were enrolled. Urinary iodine concentration (UIC), drinking water iodine content (WIC), thyroid-stimulating hormone (TSH), free triiodothyronine (FT3), free thyroid hormone (FT4), thyroglobulin (Tg), serum thyroid peroxidase antibody (TPOAb), and thyroglobulin antibody (TgAb) were measured. Pregnant women in both Shandong and Tianjin were iodine sufficient, but the median UIC in pregnant women was significantly higher in Shandong (244μg/L) than thatin Tianjin (159μg/L). No differences were found in UIC over the course of gestation in Shandong. In Tianjin, the UIC decreased during 13-24weeks and stabilized thereafter. Compared with Tianjin, TSH levels were higher and FT3 and FT4 levels were lower in Shandong. Both FT3 and FT4 significantly decreased during pregnancy in Shandong and Tianjin. TSH and Tg increased over the course of gestation in both Shandong and Tianjin. The iodine status of pregnant women in Tianjin and Shandong were sufficient, but different changing patterns in UIC and thyroid function during pregnancy were presented. More attention should be focused on iodine nutrition of pregnant women, even in iodine-sufficient areas.