The State Department said Chinese entities kept supplying missile parts to countries of concern and China had continued to develop its "biotechnology infrastructure" and "engage in biological activities with potential dual-use applications".
Chinese Foreign Ministry spokesman Hong Lei said China had made important contributions towards nonproliferation.
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"In recent years, the United States has in many venues offered full approval for China's efforts and contributions made," he told a daily news briefing.
But the latest State Department report had "played the same old tune" and "made presumptuous comments", Hong added.
"China is astonished and most dissatisfied. The comments by the United States are totally baseless. China is resolutely opposed to them," he said, adding that this is not the way to go about getting cooperation between countries on this issue.
While China has signed up for tough UN sanctions on Iran and North Korea, which both have controversial nuclear program, Chinese companies occasionally come under suspicion for selling them both banned weapons and parts.
China says it is committed to enforcing sanctions on both countries.
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