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31.
One of the consequences of increased intracellular calcium in response to a variety of physiological stimuli is the calcium activation of cytosolic proteases. Unlike lysosomal proteases with broad specificity, these calcium-activated neutral proteases show limited proteolysis of a restricted set of substrate proteins suggesting they may play a regulatory role in cellular physiology. In this study we show that the neural cell adhesion molecules NCAM-180 and N-cadherin are substrates for such endogenous calcium-activated neutral proteases. In contrast, a third neural cell adhesion molecule G4/L1 was not susceptible to calcium-activated proteolysis. The threshold for activation of NCAM and N-cadherin proteolysis is in the micromolar range of calcium suggesting that NCAM and N-cadherin are substrates for a mu-type calpain (calpain I). The site recognized by this protease is within intracellular domains of NCAM-180 and N-cadherin which are important for their interaction with cytoskeletal components. These results suggest that calcium-activated proteolysis at these sites in vivo could disrupt the linkage between extracellular ligand binding to these adhesion molecules and the normal intracellular effectors of such extracellular binding events. 相似文献
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The effects of intracellular Ca2+ (Ca2+i) on K+ currents in hippocampal cells were examined using acutely isolated cells obtained from adult guinea pigs. Whole-cell voltage-clamp recordings were carried out in a configuration that allowed a continuous perfusion of the intracellular medium. Recording media were made to block inward currents and allowed selective activation of K(+)-dependent outward currents. Voltage-dependent outward currents consisted of an initial rapidly decaying component followed by a sustained component. The time constant of decay of the transient current was about 25 msec, and previous studies (Numann et al., 1987) showed that the kinetic and pharmacological properties of this current closely resembled the A current recorded in invertebrate neurons (Connor and Stevens, 1971; Thompson, 1982). Intracellular perfusion of hippocampal cells with a solution containing elevated Ca2+ (about 4.5 x 10(-4) M) elicited outward currents at the holding potential (-45 to -55 mV) and produced changes in voltage-dependent K+ currents. The transient outward current (IA) activated by depolarization was suppressed with increases in Ca2+i. Delayed, sustained K+ currents were greatly potentiated. Data also showed that, among the 3 effects elicited by Ca2+i, suppression of IA was most sensitive to Ca2+i elevation. Previous results (Numann et al., 1987) showed that IA had a lower threshold (about -45 mV) than sustained currents (about -40 mV). By using low levels of depolarization (-40 mV), IA can be selectively activated, and the suppressive effect of Ca2+i on IA was confirmed on the kinetically isolated IA.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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The effect of the traditional Chinese medicinal herbs enema and enteric-coated capsules in the treatment of ulcerative colitis (UC) were compared in 260 cases. The immune complexes and the dynamic change of autoantibodies were monitored in 28 out of the 260 cases before and after treatment. The following results were observed. (1) There was no significant difference in the total effective rate between the enema group and the oral capsule group (93.3% and 87.5% respectively), but the recovery rates of purulent hemafecia, mucusfecia and erosion accompanying colitis, etc. in the former group were higher than those in the latter (P less than 0.01). (2) The circulating immune complexes were found 43 times above the normal range in 17 cases with positive rate 60.7%, and tended to decrease as the condition became better after treatment. Antinuclear antibodies were determined by the indirect fluorescent immune method and the indirect enzyme labelling method and the positive rates were 53.6% and 64.7% respectively, both being much higher than those in the controls (P less than 0.01). 相似文献
36.
人肿瘤特异抗原MAGE-3基因疫苗的构建和表达 总被引:8,自引:1,他引:7
目的:克隆肿瘤特异性共享抗原mage-3基因,制备基于α-病毒复制酶的高效黑色素瘤特异性基因疫苗。方法:用RT-PCR方法从黑色素瘤细胞系LB373中制备mage-3基因,以含α-病毒复制酶的哺乳细胞高效表达质粒pSMART2a为载体,构建重组DNA疫苗。重组子用载体中的T7和T3启动子序列为测序引物进行自动测序。再将鉴定过的重组质粒用脂质体法转化293细胞,用免疫印迹法和免疫组化法鉴定转化细胞中mage-3蛋白的表达。结果:正确构建了mage-3/pSMART2a重组质粒,并且在转化此质粒的293细胞中检测出了mage-3蛋白的表达。结论:此重组mage-3/pSMART2a质粒可以作为肿瘤特异的DNA疫苗,可进行下一步的肿瘤动物模型的疫苗接种及相关免疫学效应研究。 相似文献
37.
成人成骨细胞与珊瑚羟基磷灰石的体外生物相容性 总被引:13,自引:2,他引:11
目的:研究成人骨髓成骨细胞与珊瑚羟基磷灰石(carolline hydroxyapatite,CHA)在体外培养条件下的生物相容性。方法:抽取健康成人骨髓组织,置于含体积分数为10%胎牛血清的DMEM培养基中培养,传代后改用含地塞米松、β-甘油磷酸钠和维生素C的条件培养基培养,分为CHA复合细胞组和单纯细胞组,不同时间用倒置相关显微镜,HE染色光镜及扫描电镜观察,MTT法进行细胞增殖测定,并进行细胞微量蛋白含量和碱性磷酸酶的定量检测,结果:成人骨髓成骨细胞体外培养时复合或不复合CHA均生长良好,表现出典型成骨细胞的形态特征和生物学特性,CHA利于细胞的贴附,生长与增殖,并对细胞的功能无不良影响,结论:CHA是较理想的骨组织工程支架材料,成骨细胞复合CHA用于骨缺损的修复,具有广阔的临床应用前景。 相似文献
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目的总结和探讨脑血管造影和载瘤动脉闭塞在治疗颅内巨大动脉瘤上的作用及特点。方法60例颅内巨大动脉瘤患者,根据其脑血管造影的特点采取血管内介入方法[可脱式球囊和(或)弹簧圈]闭塞载瘤动脉近端53例、闭塞载瘤动脉两端后孤立动脉瘤7例;其中23例闭塞前先行颅内-外血管搭桥术。结果出院时Rankin评分分级:单纯血管内介入治疗组37例中轻残3例,合并颅内-外血管搭桥术组23例中死亡1例、重残2例。1-6年的影像学随访动脉瘤无复发。结论血管内介入结合颅内-外血管搭桥术闭塞载瘤动脉是治疗颅内巨大动脉瘤的方法之一。 相似文献
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