ralph lauren outlet Oxymatrine on protective effec

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1.2 Main instruments and reagents constant cold box frozen microtome (LEica, Germany) inverted fluorescence microscope (Nikon, Japan), UV spectrophotometer (CE2041,[link widoczny dla zalogowanych], Beijing). Reagents: SOD, MDA kit from Nanjing Jiancheng Bioengineering Institute; step TUNEL Apoptosis Detection Kit Pik days by Jiangsu Institute of Biotechnology.

Key words cerebral infarction; oxymatrine; protection

1.1 experimental animals and healthy adult male mice group 45, weight 35 ~ 40 g, Changchun Medical College Animal Center, were randomly divided into sham operation group, model group and OMT pretreatment, n = 15.
Oxymatrine on protective effect in mice with focal cerebral infarction

【Abstract】 Objective Oxymatrine (OMT) in mice after focal cerebral infarction (CI) in rats. Method of adult male mice were randomly divided into sham operation group, model group and the OMT pretreatment group. Patching method in mice with permanent middle cerebral artery occlusion model. Sham operation group and model group were injected intraperitoneally 30 min before surgery of 0.9% saline 4 ml, OMT group injected OMT 35 mg / kg. After the mice awake neurologic impairment, TTC staining infarct volume, SOD activity of xanthine oxidase method, TBA determination of MDA content, TUNEL method for the determination of apoptosis. Results Compared with model group, OMT group of mice 24 h after surgery decreased neurological function, reduced infarct size, SOD activity increased, MDA levels decreased neuronal apoptosis reduced. Conclusion OMT can play by increasing the SOD activity in mice with focal CI protection. Chinese papers League finishing.
1.3 model making and administration method of permanent middle cerebral artery occlusion in mice model making reference 〔3〕. Mice with 10% chloral hydrate (300 mg / kg) intraperitoneal injection of anesthesia, supine fixed, conventional skin preparation, the neck incision after disinfection, isolation and exposure of the right common carotid artery and external carotid artery, proximal common carotid artery ligation end of the beginning part of carotid artery, carotid artery near the bifurcation sets of alternate lines, a temporary clip occlusion of artery of internal carotid artery. Carotid bifurcation at a distance of 2 mm at the shear around a small mouth, insert the single strand of nylon cord (length 3 cm,[link widoczny dla zalogowanych], diameter 0.104 mm, the separation of markers at 1,2,3) about 11 mm, a slight resistance when stopped, truss sutures, cut excess suture, suture and re-disinfect the skin. In addition to the sham group did not insert lines, the remaining steps the same. Technique using heat to maintain mouse rectal temperature incandescent about 37 ℃. OMT plants in Shaanxi Hui Development Co., Ltd., purity 98%, production batch SF20061215, When you use the distilled water into suspension. OMT pretreatment group were injected intraperitoneally 30 min before surgery OMT 35 mg / kg. The mice were sacrificed 24 h after surgery.

Oxymatrine (OMT) have anti-inflammatory, antioxidant, anti-virus and other aspects of immune regulation 〔1,2〕 pharmacological effects, has been used for viral and immune regulation to therapy, but there is no attempt to cerebrovascular disease. The experimental production of mice with permanent middle cerebral artery occlusion model of cerebral infarction observed OMT (CI) mice with neurological deficits, infarct volume, superoxide dismutase (SOD) activity, malondialdehyde (MDA), and neuronal apoptosis and to explore whether the OMT has a protective effect on the CI and its mechanism.
1 Materials and methods

1.4 OUTCOME MEASURE (1) neurological deficit scores: a clear neurological deficits in mice after the rate 〔4〕. 0 for the non-neurological deficiency symptoms; one can not fully extend the left forepaw; two to gently pull back and hind tail lift, the left forepaw was significantly lower grip; 3 for the freedom of movement, but the traction tail will tilt to the left, or turning in circles; 4 to tilt or circling movement to the left; 5 for the stimulus movement; 6 to stimulate the free movement, accompanied by disturbance of consciousness; 7 to death; 0 to 7, respectively, 0 to 7 points. Select 1 to 5 corresponding index test mice,[link widoczny dla zalogowanych], n = 15. (2) red tetrazolium (TTC) staining infarct volume: 3 rats in each group were 5, 24 h after surgery brains were removed quickly, -20 ℃ 20 min underwent frozen coronal sections 1 mm thick and immediately placed in 1% TTC solution at 37 ℃ in the dark were incubated 30 min, normal brain tissue red, white infarction. Stained sections were scanned and access to computers. Image  Pro with image analysis software to calculate the percentage of infarct volume. (3) SOD activity and MDA content determination: each group from each of five mice brains were removed quickly, the ice tray isolated on the right frontal and parietal brain tissue, dry filter paper moisture, weighed and made into 10% tissue homogenate,[link widoczny dla zalogowanych], 4 ℃ 3 500 r / min 10 min, the supernatant centrifugation, Coomassie brilliant blue protein determination, according to kit instructions. (4) TUNEL method for the determination of apoptosis: from each of five mice in each group, anesthetized 24 h after rapid thoracotomy exposed the heart, the left ventricle to ascending aorta cannulation, right atrial appendage cut, first 0.1 mol / L pre-cooled PBS 100 ml infusion to clear, then pre-cooled 4% paraformaldehyde perfusion 150 ml. Brains were removed, 37 ℃ 4% paraformaldehyde fixed after 4 h, water rinsing overnight, 5%, 10%, 15%, 20%, 25%, 30% sucrose gradient dehydrated until the sink to the bottom, to do with the frozen section machine consecutive coronal slices, slice thickness of 6 μm. Sections were dried at room temperature, containing 1% Triton X 100 in PBS washed 3 × 5 min, TUNEL detection kit according to the instructions prepared test solution, add 50 μl to the sample test solution, 37 ℃ dark incubated for 60 min, incubated to 45 min stained with DAPI was added. PBS dark wash 3 × 8 min, after the chip seal was observed under a fluorescence microscope, the cells were positive. N = 5 slices, each slice to take five non-overlapping horizons calculated the percentage of positively stained cells. Normal nuclei in blue, yellow nucleus of apoptotic cells in green.

1.5 statistical measurement data to x ± s said that the single factor analysis of variance.

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