Using fura-2, a fluorescence indicator, we evaluated the changes of platelet cytosolic free calcium concentration ([Ca2+]i) and its role in regulation of thromboxane B2 (TXB2) synthesis in patients with diabetes mellitus (DM group, n = 27) and in healthy subjects (control group, n = 15). The A23187-evoked elevation of [Ca2+]i and TXB2 production were higher in the DM group than in the control group (P < 0.01 respectively). The rise in [Ca2+]i was correlated positively with TXB2 production. In contrast, stimulation with arachidonic acid, TXB2 production was unaltered between the groups, although arachidonate-induced [Ca2+]i was higher in the DM group than in the control group (P < 0.05). The results suggested that changes in platelets [Ca2+]i in diabetic subjects may contribute to increase in TXB2 synthesis, involved libration of free arachidonate from membrance phospholipids by the action of phospholipases. Since no significant difference was found between the diabetic patients with microangiopathy (n = 13) and without microangiopathy (n = 14), the results above may be involved in the development of diabetic microangiopathy.