Use of a radioimmunoassay to quantify thrombospondin

Results of radioimmunoassay procedures applied to samples containing thrombospondin indicated that reliable values are obtained either in saline or in plasma. Plasma levels in apparently normal individuals ranged from approximately 20 to 300 ng/ml. The mean for 20 individuals was 175 ng/ml. Plasma specimens stored either refrigerated at 4 degrees C or frozen at -80 degrees C showed significantly diminished thrombospondin levels over a period of 90 days. Serum levels of thrombospondin were found to range from 10,000 to 30,000 ng/ml.     

缺血预处理程序中心肌环磷酸腺苷含量及环磷酸腺苷依赖蛋白激酶活性的变化

目的:制备大鼠在体缺血再灌注模型,观察缺血预处理程序中心肌环磷酸腺苷含量及环磷酸腺苷依赖蛋白激酶活性的变化。方法:实验于2005-03/2006-10在解放军沈阳军区总医院医学实验动物中心和全军心血管研究所实验室完成。实验分组:选用健康雌性SD大鼠36只,根据预适应程序分为第1,2,3次缺血,第1,2,3次再灌注,每一时间点6只大鼠。实验过程:用手术套管法造成左冠状动脉主干缺血及再灌注。所有实验动物在实验程序结束后,取出心脏迅速置液氮保存备用。实验评估:用放射免疫法测环磷酸腺苷水平,生化法测环磷酸腺苷依赖蛋白激酶活性变化。结果:36只大鼠均进入结果分析。①环磷酸腺苷含量:第1次再灌注组低于第1次缺血组[(0.325±0.015),(0.395±0.024)pmol/g,t=6.06,P<0.001],第2次再灌注组低于第2次缺血组[(0.523±0.017),(0.708±0.067)pmol/g,t=6.56,P<0.001],第3次再灌注组低于第3次缺血组[(0.567±0.031),(0.712±0.038)pmol/g,t=7.24,P<0.001]。②环磷酸腺苷依赖蛋白激酶活性:第1次再灌注组低于第1次缺血组[(10.115±1.000),(16.351±0.849)pkat/g,t=11.12,P<0.001],第2次再灌注组低于第2次缺血组[(11.877±2.213),(14.869±0.619)pkat/g,t=3.31,P<0.01],第3次再灌注组低于第3次缺血组[(11.745±0.987),(14.766±0.329)pkat/g,t=7.09,P<0.001]。③缺血预处理程序中心肌环磷酸腺苷含量及环磷酸腺苷依赖蛋白激酶活性随缺血及再灌注呈周期性波动。在5min缺血预处理时表现为明显增高,而在间隔的再灌注程序中恰呈相反改变,有明显下降的趋势。结论:环磷酸腺苷及环磷酸腺苷依赖蛋白激酶的周期性波动变化可能是激发心肌缺血预处理的机制之一,环磷酸腺苷可能在预处理保护作用中起一些作用。     

Reflex penile erection in anesthetized mice: an exploratory study

Ejaculatory-like rhythmic contractions of the bulbospongiosus (BS) muscle and penile erection can be elicited in the urethane-anesthetized rat, following spinal cord transection, upon electrical stimulation (ES) of the dorsal penile nerve (DPN). The aim of this work was to investigate this reflex in anesthetized mice. Adult C57BL6 mice were anesthetized with isoflurane. The BS muscle and corpus cavernosum were instrumented to allow quantification of the BS muscle electromyographic activity (BS EMG) and intracavernosal pressure respectively. The femoral artery and jugular vein were catheterized to allow measurement of blood pressure and compound administration. ES of the DPN, via bipolar silver electrodes, reliably evoked erectile responses in mice with intact spinal cords. The overall amplitude of the erectile response was frequency- and pulse duration-dependent. Erectile responses were abolished by bilateral cut of the sensory branch of the pudendal nerve. Transection of the spinal cord potentiated the erectile responses and increased the area under the curve of the BS EMG when compared with those animals with intact spinal cords. However, no coordinated rhythmic contractions of the BS muscle during or after the ES could be observed, with or without spinal transection. Melanotan-II failed to enhance the erectile response induced by ES of the DPN, in mice with intact spinal cords. ES of the DPN in isoflurane-anesthetized mice could be a useful model in which to study the interplay between brain and spinal cord in the control of reflex penile erection, and could take advantage of knockout mice models. However, the lack of efficacy of Melanotan-II suggests that further experiments are necessary to confirm the future utility of this model. In contrast to rats, the expulsion reflex could not be reliably elicited in mice with or without spinal transection. This latter finding suggests the existence of fundamental differences in the organization of the spinal network controlling sexual reflexes between rats and mice.