Jujuhoside A (JUA) is a main component of jujubogenia exrracted from the seed of Ziziphus jujuba Mill var spinosa (Bunge) Hu ex H F Chou (Ziziphus), which is widely used in Chinese traitional metiicine for the treatment of insomnia and anxiety. Previously, we reported the inhibitinry effects of JuA on hippocampal forma-tion in vivo and in vitro, the present study was carried out to ex amine the effects of JuA on glutamate (Glu)-mediated excitatory signal pathway in hippocamput. Microdialysis coupled with high-performance Liquid chromatography (HPLC) was used to monitor the changes of Glu Levels in the hippocampus induced by penicillin sodium, or a mixture of penicillin sodium and JuA. The results showed that penicillin increased the hippocampal GlU concentration (p<0.Ol) and a high dose of JuA (0.1g/L) sig-mificantly blocked penicillin-indued Glu release (P<0.05). Moreover, the effect of JuA on intracellular Ca2+ changes after the stimulation by Glu was studied in cultured hippocampal neurons with can focal laser scanning microscope (CLSM). It was found that Clu (O.5 mM) reduced an intracellular [Ca2+], increase (p<0.01), and JuA significantly inhibited the Glu-induced Ca2+. increase. The calmodulin (CAM) antatgonist trifluoperazine (TFP) showed a similar inhibitory effect as JuA. These observations suggested tbat JuA has inhibitory effects on Glu-mediated excita-tory signal pathway in hippocampus and probably acts through its anti-calmodulin action

A method for the direct measurement of intracellular nitric oxide (NO) production stimulated by penicillin G (PG) in cultured hippocampal neurons with diaminoanthraquinone (DAA) using laser scanning confocal microscopy (LSCM) was developed. Intracellular DAA fluorescence could specifically represent NO production based on two facts: (1) 3-morpholinosydnonimine, a NO donor, could dose-dependently increase DAA fluorescence; and (2) haemoglobin, a NO scavenger, could inhibit the increase of DAA fluorescence. The PG dose-dependently increasd the intercellular level of glutamate (Glu, 5 min after stimulation) and the intracellular NO production (30 min throughout stimulation). The increase of NO production could be reversed by Nw-nitro-L-arginine (a NO synthase inhibitor), and also by D(-)2-amino-5-phosphonovaleric acid, a subtype of Glu receptor antagonist. These results revealed that DAA could be used to indicate real-time and kinetic intracellular NO production of hippocampal neurons with higher sensitivity, specificity and accuracy.

Confocal laser scanning microscopy (CLSM) is one of the common equipments being used to observe the intracellular substances like nitric oxide (NO) and calcium ions (Ca2). The intracellular substances detection relies on the proper analysis of the fluorescence images obtained by CLSM. At present, the outlines of the cells in CLSM images from fluorescence are manually drawn by judging the contrast between the object and the background. This method is subjective and may have the possibility of inaccuracy. In this paper we have designed a technique based on digital imaging processing to automatically detect the outlines or contours of the cells and assess the production of NO labeled by fluorescence probes. The developed technique was tested on a series of images obtained from CLSM and the spread ofNO during the time course was satisfactorily estimated.

Brevlseapine is a flavonoid extracted from Engeron breviscapus. Hand. Mazz, and it has been reported that breviscapine can actirate K+ channels and block Ca2+ channels, In this paper, we stud-led the cardloprotective effects of breviscapine on elect rocardio gram (ECG) changes (ST segment elevation), infarction size in dog heart subjected to myocardial infarction caused by left co-onacy artery ligation and lactate dehydrngenase (LDH) leakage. changes of [ntracellular free Ca2+ levels, apoptosis and necrosis in cultured neonatal rat cardiomyocytes subjected to hypoxia. Adidonally, the effect of breviscapine on myocardial oxygen consumption was detected in dog myocardium in vitro. The re suits showed that breviscapine treatment (1mg/kg, 2 mg/kg and 4mg/kg) significantly reduced ST-segment elevation and in-farction size in hearts subjected to myocardial infarction, that breviscapine treatment (1429ug/mL, 28.57ug/m L and 57.14ug/mL) significantly decreased oxygen consumption in myoear-dium, and that breviscapine treatment (5 Ug/mL. 10Ug/mL and 20Ug/mL) significantly reduced LDH leakage, intracellular free Ca2+levels, apoptosis and necrosis in cardiomyocytes subjected to hypoxia. In conclusion, the present study indicates that blevis-capine is in favor of myocardial protection.

A novel method based on flow cytometry (FCM), which can count the number of detected cells, has been developed for the evaluation of cellular proliferation and cytotoxicity in vitro. It provides a tool that directly counts cell number without being influenced by the metabolic state of the cells, discriminates target cells from effector cells in cell-mediated cytotoxicity assay, and with less treatment step and free radioactivity. In this paper, we have prepared the PG cells (a highly metastatic human lung cancer cell line) and peripheral blood lymphocytes (PBL) with various concentrations and ratios of concentration to validate the method. The results were compared with MTT (3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide) assay and the regression analysis results showed that this method worked very well. We have also used this method to evaluate mitogeninduced proliferation and cytotoxicity. The results indicated that this method might yield high sensitivity and reliability.