In this work, we present a systematic study on the complete control over the sphere diameters of MCM-41 nanospheres with different pore sizes. The mesoporous silica nanospheres with diameter range from 40 nm to 160 nm are synthesized by using cationic quaternary ammonium surfactants as template and TEOS as silicon source in sodium hydroxide aqueous solution via sol-gel supermolecular method. Nanospheres with fixed diameter can be obtained through the precise control over the molar ratio of NaOH/TEOS and other synthetic conditions. The range of sphere diameter for each sample prepared under adjustable reaction conditions is in a sharp distribution. Moreover, the formation mechanism for nanospheres with controlled morphological and structural features is proposed, and we suggest the optimal synthesis conditions of the MCM-41 nanospheres for the minimization of the Gibbs free energy deltaG. Besides, after depositing vanadium species on the MCM-41 support, the catalytic performance of V-MCM-41 for the selective oxidation of styrene by hydroperoxide increases with the diameter of the supports decreasing.