On the basis of the first oxide prepared anodized aluminum oxide template (AAO), the second, third and fourth oxidation preparation are implemented in turn. Then, the surface morphology and current curve changes of the AAO template with multiple oxidation preparation were compared and analyzed systemically. The apertures of nanopore made by the second, third and forth oxidation increase in order, the intervals between neighboring apertures reduce. However, the scale of ordered distribution of nanopores on the whole template surface does not change significantly. Then, carefully controlling the working conditions for the membrane-removed of the one-step oxidation AAO, the nanoporous with regular apertures and highly ordered was obtained after getting rid of the top oxidation layer about 10 minutes. Finally, based on those prepared AAO templates, integrated with the thermal nanoimprint lithography, we achieved the PMMA nano-pillar arrays with controllable lengths and diameters.