The significance of the neostigmine test is that it tests for dysfunction of the neuromuscular junction, and a positive test indicates that the patient is more likely to have myasthenia gravis. The neostigmine test is usually performed as follows: neostigmine 0.5-1mg intramuscularly, along with atropine 0.5mg, in order to avoid side effects of neostigmine, such as abdominal pain, diarrhea, vomiting, bradycardia, etc. If the neostigmine and atropine are given within 30 minutes of each other, the neostigmine and atropine test will not be effective. A positive neostigmine test result is indicated if there is a significant reduction or relief of the patient’s muscle weakness symptoms 30 minutes after the injection of neostigmine and atropine. Positive neostigmine test indicates that the patient’s neuromuscular junction transmission is impaired, and if the injection of neostigmine improves the transmission between the neuromuscular junction, and the patient’s muscle strength increases, the patient is more likely to be a patient with myasthenia gravis, but it is necessary to combine with the heavy-frequency electrical stimulation, acetylcholine receptor antibody test and other tests to assist in the diagnosis.