异氟醚预处理对大鼠脑缺血/再灌注损伤细胞内游离钙的影响

目的:观察异氟醚对缺血再灌注大鼠活体脑片细胞内游离钙的影响,探讨异氟醚脑保护作用的机制。方法:采用线栓法阻闭大鼠大脑中动脉造成局灶性脑缺血,并于缺血2h后拔出栓线给予再灌注。利用细胞内钙离子荧光探针Fluo-3/AM在激光共聚焦显微镜下观察再灌注2h后大鼠脑片细胞内荧光强度。比较异氟醚预处理1h对缺血再灌注大鼠活体脑片额顶叶皮质及纹状体区域Ca2 浓度的影响。结果:缺血再灌注损伤大鼠缺血侧脑片皮层及纹状体Ca2 荧光强度明显升高。脑缺血前给予异氟醚预处理1h可抑制缺血侧大脑皮层细胞内Ca2 含量的增加,对纹状体细胞内Ca2 含量的影响无统计学差异。结论:缺血前给予异氟醚预处理可有效减轻额顶叶皮层细胞内的钙超载,对纹状体区域脑细胞内游离钙水平影响较小,提示异氟醚的脑保护作用在不同脑区存在差异。     

Scanning Electron Microscopic Evaluation of Nonocclusive Excimer Laser-Assisted Anastomosis in Rabbits

Summary  Background. The nonocclusive Excimer laser-assisted bypass technique has been described in previous studies and proved to be a promising bypass operation in vascular brain surgery. Little is known about the morphological regeneration process of the laser-assisted anastomosis in time.  By way of a scanning electron microscopic study we examined the way in which the anastomosis site created by the nonocclusive Excimer laser-assisted anastomosis technique becomes endothelialized.  Methods. In 14 rabbits the internal jugular vein was placed in a loop on the abdominal aorta. The distal anastomosis was made using the nonocclusive Excimer laser-assisted technique. The proximal anastomosis was made either laser-assisted or conventional end-to-side. After clipping of the aorta between the two anastomoses sites the vein served as a bypass. To evaluate the endothelialization at the laser-assisted anastomosis site in time, a scanning electron microscopic study was performed.  Results. In the first hours after the bypass operation a new intimal surface is formed by fibrin and activated platelets. Some leukocytes are seen during the first days. The endothelialization process of the laser-assisted anastomosis site begins one day after the operation. The gradual endothelialization process evolved along two lines. First, endothelial cells grow from the side of the aorta to the bypass. Second, after one day solitary (blood-borne) endothelial cells deposit on the laser edge and the sutures, covering the platelet aggregates.  Conclusions. The endothelialization of the Excimer laser-assisted anastomosis is more or less completed 9 days after the operation. The edge created by the laser becomes smoother after a few days and is gone for the most part after 9 days.     

氩激光对培养的牛视网膜色素上皮细胞损伤的扫描电镜观察

目的采用扫描电镜观察氩激光对体外培养的牛视网膜色素上皮(RPE)细胞表面的损伤情况。方法培养牛视网膜色素上皮细胞,将第4代细胞传代成融合状态后,用氩激光对RPE细胞进行照射,激光能量为0.8W,曝光时间为0.2s,光斑直径为200μm,激光照射后用扫描电镜观察色素上皮细胞表面超微结构变化。结果激光照射后,可见光斑中央区的细胞脱失,光斑周边损伤细胞主要表现为表面微绒毛的减少或消失,微绒毛卷曲,有些细胞边缘卷曲及细胞膜破裂出现不规则小孔。结论激光对体外培养的RPE细胞照射可造成色素上皮细胞脱失,细胞膜损伤及微绒毛损伤。     

玉米赤霉烯酮对细胞间隙连接通讯的影响

目的 了解真菌性雌激素玉米赤霉烯酮(ZEA)对细胞间隙连接通讯(GJIC)的影响。方法 用荧光漂白后恢复(fluorescence redistribution after photobleaching，FRAP)技术，在激光扫描共聚焦显微镜下观察ZEA对HaCaT细胞GJIC功能的影响。结果 ZEA在0．1μmol/L对HaCaT细胞的GJIC没有明显影响，但在I—100μmol/L浓度下都有明显的抑制作用。ZEA不能有效地拮抗TPA引起的GJIC抑制。结论 ZEA在1μmol/L以上就能抑制GJIC功能，提示它在一定条件下可能有促癌作用。     

Experimental Study on Meiotic Spindles and Chromosomes of Mouse Mature （MⅡ） Stage Oocytes under Laser Scanning Confocal Microscopy

The in vitro culture,maturation and cryopr-eservation of oocytes are all the i mportant parts inthe assisted reproductive technology.Mature oo-cyte(i.e.MⅡoocytes)is arrested at the meta-phase stage of meiosisⅡ,in which chromosomesare attached to the spindle microtubles and line uponthe equatorial plateinthe normal spindles.Dis-organized structure of spindles can not onlylead tosuch morphologic changes as unwound chromo-somes but also the chromosome nondisjuction andresultant aneuploid.When t…     

N-苄基氟哌啶醇氯化物对大鼠血管平滑肌细胞内钙离子的影响

目的:动态观察N_苄基氟哌啶醇氯化物(F3)对大鼠胸主动脉平滑肌细胞内Ca2+的影响。方法:用Ca2+荧光染料Fluo_3_AM负载大鼠胸主动脉平滑肌细胞,在激光扫描共聚焦显微镜上测定细胞内游离钙的变化。结果:在含CaCl2(1.26mmol/L)的细胞外液中,30 mmol/L KCl使细胞内钙荧光强度(FI)迅速增强,F3可以拮抗KCl诱导钙FI增强作用,并呈量效依赖和时间依赖关系。4种浓度F3(0.01、0.1、1.0、10μmol/L)作用3 min时,使KCl诱导细胞内钙FI从(81±4)%分别降至(71±18)%(n=20,P<0.05)、(51±12)%(n=20,P<0.01)、(39±14)%(n=20,P<0.01)、(28±14)%(n=20,P<0.01)。钙FI变化最快的时程是在给药后的0~30 s。结论:F3通过阻断细胞膜电压依赖性钙通道降低平滑肌细胞内钙浓度,这可能是其扩张血管,改善心肌缺血的机制。     

正交法选定肾泰安胶囊的提取工艺

目的：选定肾泰安胶囊植物药部分的提取工艺。方法：采用正交试验法和方差分析，对肾泰安胶囊植物药部分的醇提工艺从药材粉碎度、回流时间、乙醇加入量等方面进行选择；水煮工艺从煮提时间、加入水量等方面进行选择。结果：将药材饮片打碎，加入3倍量80％乙醇回流1h为最佳醇提条件；残渣用8倍量水煎煮40min为最佳水提条件。结论：所选定的工艺是肾泰安胶囊剂生产中的最佳提取工艺。     

Serial changes in N-isopropyl-p[125I]-iodoamphetamine in mouse lung observed with a confocal laser scanning microscope

Serial changes in N-isopropyl-p[125I]-iodoamphetamine (125I-IMP) in mouse lungs were observed with a confocal laser scanning microscope. Male mice were intravenously injected with 125I-IMP and subjected to autoradiographic procedures 20 minutes, and 3 and 24 hours after injection. Differential interference contrast (DIC) images and confocal images were obtained with a confocal laser scanning microscope, and superimposed images were evaluated. Large numbers of silver grains were observed in the interstitium, bronchioles, and alveolar sacs 20 minutes after the injection, and lamellar distribution of the grains was observed on the ciliary surface. The numbers of silver grains in the interstitium and bronchioles had decreased 3 hours after the injection of 125I-IMP, but the numbers of silver grains in the alveolar spaces had not. Although small numbers of silver grains remained in both the bronchioles and alveolar sacs 24 hours after the injection, most of them had washed out. Confocal laser scanning microscopy is considered to be a useful procedure for studying the distribution of radioisotopes by microautoradiography, because it allows clear autoradiographs to be obtained in which tissues and silver grains are perfectly matched and all silver grains are in focus.     

薄层扫描法测定血尿康颗粒中盐酸水苏碱的含量

目的:建立以薄层扫描法测定血尿康颗粒中盐酸水苏碱含量的方法。方法:采用硅胶G薄层板,展开剂为正丁醇-盐酸-醋酸乙酯(8∶3∶1),显色剂为稀碘化铋钾试液,测定波长为515nm,参比波长为700nm,狭缝大小为6.00mm×0.45mm。结果:盐酸水苏碱点样量在5.06μg～25.32μg范围内与斑点峰面积积分值呈良好的线性关系(r=0.9978),平均加样回收率为96.67%(RSD=2.01%)。结论:本方法简便、灵敏度高、重现性好,可用于血尿康颗粒的含量测定与质量控制。     

薄层扫描法测定垂阴茶糖浆中岩白菜素的含量

目的:建立垂阴茶糖浆岩白菜素的测定方法,方法:λs=275 nm,λR=320nm,用薄层色谱法测定岩白菜素的含量.结果:岩白菜素在0.505～3.030μg范围内线性关系良好(r=0.999 9),平均回收率为96.52%.RSD=0.44%.结论:该法灵敏度高,重现性好,可用于该制剂中岩白菜素的含量测定及质量控制.