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.

Nitric oxide (NO) and calcium ions (Ca2) play critical role as molecular mediator in many physiological processes. However, their low concentration and instability in specimen make them difficult to be detected directly in neurons. We developed a method for imaging nitric oxide and calcium ions using Laser Scanning Confocal Microscopy (LSCM). Cultured hippocampal neuron is dyed and observed under Zeiss LSM 510 laser scanning confocal microscope. Excited by laser the emission light from the labeled nitric oxide and calcium ions in the neutron are detected. In this way, the nitric oxide and calcium ions are imaged and their intracellular kinetic change is monitored. Furthermore, image processing and visualization techniques are employed to help analyze the image data.

In a photochemical reaction microvessel thrombosis model, the anti-thrombotic effect of PT was evaluated and the results sbowed that PF could significantly prolong thromobosis time. The anti-thrombotic effect of PF may relate to the inhibi-tion of arachidonic acid metabolism, the increase of t-PA activi ty, and the protective effect against free radical.

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.