Core physique tempe rature was keeping having a rectal probe and maintained at 36 37 C applying a homeothermic blanket handle unit in the course of and right after the surgical procedure operation. Twenty 6 rats un awakened or died following 2 h of operation were rejected out, though 159 productive models whose cerebral blood flow curve dropped to 30% had been brought in to the experiment and had been randomly di vided into model group and treatment method group. Orthogonal experimental design and style and intervention The treatment method group rats have been sub grouped in accordance towards the principle of orthogonal experi psychological design and style of consisting of two influence components with 4 effect ranges. The influence factor A would be the the rapeutic time widow built four ranges as 1. 0 h, 1. 5 h, 2. 0 h, two. five h soon after ischemia.
The influence issue B could be the therapeutic drug dose which has four ranges as following 5 mg kg, 10 mg kg, 20 mg kg and 40 mg kg physique bodyweight. The orthogonal experimental test was repeated 3 times. Picroside II which the purity exceed 98% and molecular formula is C23H28O13, selleck chemicals VEGFR Inhibitor sup plied by Tianjin Kuiqing Healthcare Technologies Co. Ltd. In accordance towards the bodyweight of rats, corresponding dose of picroside II powder was taken and diluted into one ml answer by isotonic saline answer and injected intraper itoneally according on the corresponding intended doses at intended time from the orthogonal layout. Rats in management group and model group had been intraperitoneally injected the identical dose saline soon after cerebral ischemia 2 h. The brain tissue was took out to evaluate the therapeutic impact of picroside II immediately after treatment method 24 h.
Speedy green staining The rats from handle group, model group and treatment group had been randomly chosen and anesthetized by injecting intraperitoneally 10% chloral hydrate, and perfused by normal saline 200 ml and fixed by 4% paraformaldehyde resolution 200 ml successively through heart. Then the whole brain was taken out and publish fixed in 4% formaldehyde option for 2 h selleck chemicals “ and soaked in distilled water for 4 h. Soon after conven tional gradient ethanol dehydration, xylene transparent, paraffin embedding, coronal sections which has a thickness of five um had been constantly reduce forward in the posterior of optic chiasma by a microtome then adhered within the slices processed with poly lysine. The paraffin sections have been dewaxed by dimethyl ben zene and washed routinely, dyed one h in rapidly green alcohol solution at 37 C soon after handling 95% alcohol one min, and then washed by 95% alcohol and distilled water, putted in 0. 3% lithium carbonate to separate color, re stained 90s by nuclear quickly red just after washing by distilled water, lastly washed by distilled water and conventionally dehydrated by gra dient ethanol, cleared by xylene and sealed with neutral balsam.