Besides, the novel UP-M-PCR can be used in all the fields where multiplex PCR is needed, which has promising application future and is worth being popularized. Diabetes mellitus is a group of metabolic disorders that cause chronic hyperglycemia and is one of the most significant diseases in the developed world, affecting more than 170 million people. The tissue responses to diabetic conditions are varied; many are associated with oxidative stress in the cells. The improper management of hyperglycemia leads to severe complications in diabetic patients: approximately 15% of patients display impaired wound healing, causing long-term complications such as limb amputation. Skin wound repair involves a series of coordinated processes that include cell proliferation and migration, collagen deposition and remodeling, wound contraction, and angiogenesis. These processes involve different cell types, mostly fibroblasts/myofibroblasts, keratinocytes, and endothelial cells. While hyperglycemia has been linked to impaired wound healing, particularly altered angiogenesis and extracellular matrix remodeling, the nature of the linkage is unclear. Some studies have described alterations in cell migration associated with diabetic conditions. For example, Lerman et al. showed that fibroblasts from diabetic mice migrate less than those from normoglycemic mice and display a defective response to hypoxia, a condition commonly present in chronic wounds. A similar inhibition was recently observed in keratinocytes cultured in a high Z-VAD-FMK glucose environment, which suggests that high glucose plays a direct role on cell migration. However, none of these studies addressed the cellular mechanism by which this happens. The migratory process is a cycle comprised of distinct. These steps are: polarization, in which the cell develops a clear front and rear; protrusion, which is driven by actin polymerization at the leading edge; the formation of substrate adhesions that serve to stabilize protrusions and generate the dynamic signaling, which converge on Rho GTPases. The cycle is completed with retraction at the cell rear and the release of adhesions. The small Rho GTPases are central regulators that integrate and drive these processes; they act through several effector proteins that mediate migration. For example, Rac1 regulates the formation of the lamellipodium and adhesion dynamics, while RhoA is involved in the formation of actin bundles and adhesion maturation. This study addresses the mechanism by which high glucose inhibits cell migration.