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Objective: This study group pre-clinical observations found that TBI (traumatic brain injury, TBI) patients with peripheral blood progenitor cells (endothelial progenitor cells, EPCs) number first and then there is the rising variation, in order to further study the peripheral blood EPCs change mechanism, are required to establish the relevant animal research platform. Purpose of this study: 1. Establish a separation of EPCs in peripheral blood of rats, labeling method, the use of flow cytometry (flow cytometer, FCM) double antibody labeling technique detects the number of EPCs in peripheral blood of rats; 2. Use of large Hydraulic rat brain trauma (Fluid-Percussion injury, FPI) model proved EPCs in peripheral blood of rats after TBI trends; 3. explore EPCs after TBI and white blood cells, platelets trends relevant for the study of peripheral blood after TBI Changes in the mechanism of EPCs provide a reference. Method: 1. Using healthy adult male WISTER of 45 rats (weighing 300-350g, rats aged 12 weeks) were divided into sham operation group (n = 10) and TBI group (n = 35). Sham-operated control rats given only 10% chloral hydrate anesthetized by intraperitoneal injection. TBI group against using hydraulic devices and small animal stereotactic instrument, were established light, medium and heavy-duty hydraulic closed head injury model in rats, mild group (n = 10) Hydraulic crackdown is 1atm, medium-sized group (n = 10) Hydraulic crackdown as 2atm, severe group (n = 15) Hydraulic crackdown as 3atm; 2.TBI after 24h, in accordance with the principle of random from each of the TBI group rats were each taken two cranial MR imaging and brain tissue stained with HE observe images and pathological features of TBI. The remaining rats were pre-injury and 24h after injury at different time points (3h, 6h, 24h, 48h, 72h, 240h, 336h) using rigid glass capillary blood collection tube (φ = 1mm) from rat medial canthal ball venous plexus Blood approximately 1.1ml, for determination of EPCs, while using a small animal blood cell detection equipment leukocytes and platelets determination; 3 using Ficoll density gradient centrifugation in peripheral blood mononuclear cells, CD34, CD133 double-antibody positive as EPCs logo, using CD34, CD133 double antibody labeling combined with flow cytometry was used to detect the number of EPCs in peripheral blood of rats. Results: In the control group, 10 rats were done at each time point corresponding indicators in peripheral blood circulation measured; Light TBI group 1 rats after three days due to strike the head wound infection was removed, and the remaining seven rats each completed time point corresponding indicators in peripheral blood circulation measured; TBI group, a medium-sized rats 48h after death in combat, and the remaining seven rats were done at each time point corresponding indicators in peripheral blood circulation measured; severe TBI group 2 died in combat after 24h, 48h after four killed in combat, in which one eye canthus because blood at serious infections were excluded, and the remaining seven rats were done at each time point corresponding indicators in peripheral blood circulation measured; experiment midway dead rats (n = 6) of the die set as the corresponding data for analysis. 1 expression in peripheral blood circulation in the presence of CD34, CD133 double-positive EPCs, this study tests showed: normal rat peripheral blood circulating EPCs was 33-61 months / 200,000 mononuclear cells with an average of 47/200 000 mononuclear cells; 2.TBI number of EPCs in peripheral blood of rats after injury decreased to below normal levels, 3h after injury and reached the lowest point, to 10-30 / 200 000 mononuclear cells in each group had no significant difference decline ; 3.TBI number of EPCs in peripheral blood of rats after injury 3-6h gradually increased to above normal levels at 6h after injury, reached the highest point, light group, the average rose to 80/200 000 mononuclear cells, medium-sized group The average rose to 68/200 000 mononuclear cells, heavy group, the average rose to 51/200 000 mononuclear cells, EPCs gradually decreased thereafter, 24h after injury EPCs in each group were reduced to normal levels; TBI rats died after peripheral EPCs in the blood after injury is also reaching the lowest 3h, 3-6h after injury, although the upward trend, but not reach the normal level, before the death of EPCs in peripheral blood showed a declining trend; 4.TBI rat peripheral blood Changes in the number of EPCs and platelets, white blood cell count were not significantly related. Conclusions: 1 using CD34, CD133 antibody labeling technology combined with dual-use flow cytometry can be quantitatively determined with good rat peripheral blood EPCs; 2. The experiments show that: after injury in patients with TBI changes in peripheral blood EPCs similar characteristics, TBI rats survived level also exist in peripheral blood EPCs decreased first, after the increase, and then gradually decreased to the normal variation; 3 more serious trauma, peripheral blood EPCs increased magnitude smaller, and after injury EPCs in peripheral blood of dead rats at levels below normal levels continue, tips: TBI EPCs in peripheral blood after sustained low level of indications of poor prognosis. (4) The experiments showed that: TBI after injury the number of EPCs in peripheral blood platelets and white blood cell count change and no significant correlation.
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