动物造模,自体白色血栓性猴局部脑缺血模型 z-bio 2023-09-26 917

　　1. Model material animal: Crab eating monkey, male, weighing 6-8kg, aged 6-8 years old, preoperative CT plain scan, excluding brain space occupying lesions; Drugs: Ketamine hydrochloride, diazepam, scopolamine, propofol, thrombin, etc.

　　2. Method of modeling: 2ml of venous blood was extracted 24 hours before surgery, centrifuged at 2000r/min for 10 minutes, supernatant and platelets were taken, centrifuged again at 2000r/min for 10 minutes, and the upper half of the supernatant was discarded. 10U of thrombin was added to the remaining part, and quickly injected into a 0.5mm diameter silicone tube to stand still. Refrigerated at 4 ℃ for backup, in order to quantify the size of the injected thrombus during surgery.

　　Preoperative fasting for 24 hours and limited water intake. Using compound anesthesia: 1ml ketamine hydrochloride+1ml diazepam+0.5ml scopolamine intramuscular injection. After the animal is quiet, relevant preoperative preparations are performed. The animal is fixed in a supine position on the interventional operating table, and skin is prepared in the lateral venous area of the lower limb. After routine disinfection, a trocar is used for venous puncture, and a three-way tube is connected for intraoperative intravenous medication and blood collection. Intraoperative anesthesia was maintained by injecting propofol, ketamine, and diazepam with a micro injection pump. Nasal feeding oxygen supply, heart rate, respiration, and blood pressure monitoring under the DSA system for intraoperative reference.

　　Take one side of the inguinal area of the monkey for routine skin preparation, disinfection, and tissue placement. Puncture the femoral artery with a trocar and insert a 4F catheter sheath. After inserting a 4F multifunctional catheter into the left internal carotid artery, replace it with a 3F microcatheter and send it to the M1 branch of the middle cerebral artery for anteroposterior lateral imaging. Observe the distribution of blood flow and branches of the middle cerebral artery, and use it as a reference to determine the blood flow status after embolization. Inject autologous white blood clots of about 15cm through microcatheters (the exact length depends on the size of the blood vessels indicated by the middle cerebral artery imaging), and perform another middle cerebral artery angiography to confirm poor vascular imaging after the M1 branch of the left middle cerebral artery. After 1 hour of embolization, CT perfusion imaging examination was performed, and a clear acute ischemic lesion was observed in the left brain. After confirming the success of the surgery, the interventional catheter can be removed and the puncture site can be locally pressed for 20 minutes. After compression and dressing, the animal will be sent back to the animal room for postoperative care. Depending on the postoperative status of the animal, mannitol and furosemide will be administered intravenously to reduce intracranial pressure. 72 hours after surgery, CT plain scan was performed again to confirm the formation of old ischemic lesions. Monitor the changes in animal neurological function daily after surgery.

　　3. Modeling principle: Autologous white blood clots cause middle cerebral artery embolism, leading to cerebral ischemia in animals.

　　4. Changes after modeling. After modeling, the thrombus was firm, and the infarcted area formed. The model animals showed varying degrees of left cerebral ischemia and right limb paralysis with neurological dysfunction.

　　The comparison of DSA angiography before and after embolization indicates poor vascular imaging after the M1 branch of the middle cerebral artery after embolization. CT perfusion imaging at 1 hour after embolization confirmed the presence of infarcted lesions. After 72 hours of embolization, CT plain scan showed low-density old ischemic lesions. Taking the brain for 2,3,5-chlorophenyltetrazolium (TTC) staining, obvious acute ischemic lesions can be observed.

　　5. Precautions: Surgical instruments should be strictly disinfected to prevent surgical infections. Surgical trauma should be minimized and sterile operations should be strictly carried out.