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Ni et al., 2020)

Autotomy that occurs long after injury may appear as pain that occurs below the level of the injury (below-level pain) [64, 65], and the late spontaneous worsening may be the result of complete deafferentation of one or several spinal segments the stimulation of the nerve plexus, or dorsal root injury [66]

Your calculated dose is accurate based on what went into the vial, but the actual active compound is lower because some percentage was destroyed by shaking

This occurs when there is an imbalance between free radicals (unstable molecules that can damage cells) and the antioxidants that neutralize them