Objective To evaluate the effects of locked plates with different screw layouts on stability of the internal fixation system for femoral shaft fracture, so as to provide guidance for clinical study on screw layout of the internal fixation system for femoral shaft fracture. Methods Orthogonal test design method was used with 4 groups of screws at 3 parametric levels. Biomechanical properties of the internal fixation system for femoral shaft fracture under 9 screw layouts were analyzed respectively by the finite element method, and 4 inspection indices, namely, the maximum equivalent stress of the plate, the screw and the femur, and the maximum axial displacement of the internal fixation system were obtained from each group of the experiment, and the orthogonal test weight matrix analysis method was conducted for data analysis. Results The influences of different screw parameters on stability of the internal fixation system varied, and the maximum weights of the 4 sets of screw parameters in stability of the internal fixation system were 13.86%, 7.57%, 7.53% and 5.91%, respectively. Conclusions The best scheme for screw layout in orthogonal test is the screw with none-fixed threaded holes in the first group, the single cortical screws with fixed threaded holes in the second group and the double cortical screws with fixed threaded holes in the third and fourth group.