Our preliminary study verified that even larger dose of donepezil, 5 mg/kg/day, does not downregulate heartbeat or blood pressure. At the end of the treatment period, the heart and quadriceps femoris muscle were excised for tests. To functionally evaluate the aftereffects of donepezil on murine angiogenesis, angiography was done using indocyanine green, which demonstrably visualize tissue perfusion within the hindlimb. After anesthesia with pentobarbital sodium, both lower extremities were shaved. The field was lit by a LIGHT EMITTING DIODE fluorescence imaging device, and ICG was admin istered intravenously. Following a bolus ICG Docetaxel price procedure, real-time imaging analysis was done using an infra-red camera and recorded with a digital camcorder for an enhanced time. Our preliminary studies identified that ICG angiography in the hindlimbs originally revealed an angiogram, followed closely by a perfusion period, once the fluorescence signals in the perfused muscle were simultaneously and homogeneously improved in both hindlimbs. After recording, the fluorescence intensity inside the hindlimb was evaluated. Consequently, to judge perfusion in a ischemic hindlimb, the involved regions were selected from both left and right hindlimbs. The fluorescence indicators of the left hindlimb in the perfusion Lymph node period were compared with those of the right, using NIH picture software, and it revealed laterality in the ICG signal intensity. Time once the signal difference between your left and right was extremely obvious was selected. 2. 3. Evaluation of the relative blood flow using fluorescent microspheres According to past studies using fluorescent microspheres, we examined the relative blood flow between the left and right hindlimbs in rats. Shortly, after anesthesia, 200 uL of green FluoSphere fluorescent microspheres was given within 1 min, accompanied by immediate sampling of blood and bilateral quadriceps femoris muscles. These samples were weighed correctly and incubated in 4 M KOH containing two weeks of Tween 80 at 70 C for 2-4 h. After measuring the fluorescence, the proportion of the fluorescence between your right and left hindlimbs, adjusted from the muscle weight, was compared. One day after surgery, an initial temperature assessment was performed in the hindlimbs using infra-red thermography. The skin temperature of the rats under anesthesia was measured 3 times. Then, Chk inhibitor for every mouse, temperature was assessed 3 times by thermography all through 30 days. The temperature distribution in the hindlimbs was standardized by each non ligated hindlimb and compared between donepezil treated and untreated mice like a reference. The laterality in temperature, represented by the relation between the right and left hindlimb temperature, was evaluated.