Novel application of Jingzhaotoxin

[0042] Embodiment 1, the in vitro thrombolytic activity of Jingzhao's toxin

[0043] 1. Preparation of fibrin plates

[0044] 1. Dilute 0.04g of human fibrinogen into 10ml of sterilized PBS buffer so that the final concentration is 4mg / ml, which is the fibrinogen solution.

[0045] 2. Dilute thrombin with 1ml of PBS buffer to a final concentration of 1mg / ml, which is the thrombin solution.

[0046] 3. Dissolve 0.1g of agarose in 10ml of PBS buffer and heat to dissolve, which is the agarose solution.

[0047]4. Add 1ml of thrombin solution to the wall into 10ml of fibrinogen solution, mix quickly and pour into 10ml of agarose solution melted at 37°C, mix immediately and pour into a disposable petri dish with a diameter of 10.5cm medium; place at room temperature for 30 minutes, transfer to a 4°C refrigerator and place for 30 minutes to obtain a fibrin plate; use a 100 μl tip to punch holes on the fibrin plate; irradiate for 10 minutes under ultraviolet light.

[0048] 2. Det...

[0063] Embodiment 3, the in vivo activity test of Jingzhao's toxin

[0064] 1. Establishment of a mouse cerebral ischemia-reperfusion model

[0065] Construct a mouse cerebral ischemia-reperfusion model (also known as a mouse middle cerebral artery occlusion model or MCAO model; the construction method was established by Longa et al. in 1989, and it is a classic method for studying cerebral ischemia at present; references: LONGA, E.Z., et al. al., REVERSIBLE MIDDLE CEREBRAL-ARTERY OCCLUSION WITHOUT CRANIECTOMY IN RATS. STROKE, 1989.20(1): p.84-91.), the steps are as follows:

[0066] (1) Anesthesia

[0067] Male Kunming mice (body weight 20-30 g, purchased from Guangzhou Experimental Medicine Animal Center) were injected with 10% (volume percentage) chloral hydrate aqueous solution, and the injection dose was 300 mg chloral hydrate / kg mouse body weight, intravenously.

[0068] (2) MCAO modeling

[0069] After 10 minutes of anesthesia in step (1), the initial perfusion was c...