Background: The aim of this study was to investigate the effect of dietary supplementation of fine ground corn and soybean oil on ruminal fermentation, milk performance and fatty acid profile, plasma metabolites and oxidative stress parameters in lactating dairy cows. Methods: Eight primiparous Holstein cows (215±34 d days in milk, 574.6 ± 22.6 kg body weight; mean ± SD) were allocated into 2 groups (n=4/group), used in a change-over experiment with two 23-d experimental periods (16 d induction period and 7 d wash-out period). During the induction period, the cows of one group were fed with control diets (CON, 23.8% starch, 4.6% fat, and 31.4% NDF, DM basis), and the cows of the second group were fed with CON diets supplementation of 266 g/kg of fine ground corn and 46 g/kg of soybean oil (HSO, 31.4% starch, 7.8% fat, and 26.4% NDF, DM basis), and all cows were fed with the CON diets during the subsequent wash-out period. Results: The results shown that the cows fed the HSO diets had a lower ruminal molar ratio of acetate (P = 0.02), and a numerically lower ruminal pH (P = 0.13) and acetate : propionate ratio (P = 0.13) than that in the cows fed the CON diets. Cows fed the HSO dites had a lower (P < 0.05) dry matter intake (DMI), milk yield, milk fat percentage, the yield of milk fat, protein and solid of non-fat (SNF), and a higher (P<0.05) percentage of milk protein, lactose and SNF than that in the cows fed with CON diets. Cows fed the HSO diets had a lower (P < 0.05) concentrations [g/100 g total FA] of C8:0, C10:0, C11:0, C12:0, C14:0, C15:0, C16:0, C17:0, C18:0, fatty acids (FA) <C16, sum of C16:0 and C16:1, total saturates FA, and medium chain FA, and a greater (P < 0.05) milk concentrations of C14:1, C16:1, C18:1n9t, C18:2n6t, C18:2n6c, C18:3n3, FA > C16, total unsaturated FA, total mono-unsaturated FA, total poly-unsaturated FA, Δ9-desaturase index, than that in the cows fed the CON diet. Compared with the cows fed the CON diet, the concentrations of plasma non-esterified fatty acids (NEFA), aspartate aminotransferase (AST), malondialdehyde (MDA) increased (P < 0.01), whereas the concentrations of theβ-hydroxybutyrate (BHBA), urea nitrogen (BUN), alanine aminotransferase (ALT), albumin (ALB), total antioxidant capacity (T-AOC), the ratio of ALB to GLB, the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) decreased (P < 0.05) in cows fed the HSO diets. Conclusions: Our results indicate that HSO diets can impact on the ruminal fermentation pattern, reduce the DMI, milk fat content, depress the de novo synthesis of lipid in mammary gland, disturb plasma parameters, enhance the oxidative stress in dairy cows. Meanwhile, we also found the the subacute rumen acidosis (SARA) and milk fat depression (MFD) can occur at the same time or one after the other, when dairy cows fed with HSO diets.