Assessment of Neuronal Viability Using Fluorescein Diacetate-Propidium Iodide Double Staining in Cerebellar Granule Neuron Culture.

Primary cultured Cerebellar Granule Neurons (CGNs) have been widely used as an in vitro model in neuroscience and neuropharmacology research. However, the co-existence of glial cells and neurons in CGN culture might lead to biases in the accurate assessment of neuronal viability. Fluorescein diacetate (FDA) and Propidium Iodide (PI) double staining has been used to measure cell viability by simultaneously evaluating the viable and dead cells. We used FDA-PI double staining to improve the sensitivities of the colorimetric assays and to evaluate neuronal viability in CGNs. Furthermore, we added blue fluorescent DNA stains (e.g., Hoechst) to improve the accuracy. This protocol describes how to improve the accuracy of assessment of neuronal viability by using these methods in CGN culture. Using this protocol, the number of glial cells can be excluded by using fluorescence microscopy. A similar strategy can be applied to distinguish the unwanted glial cells from neurons in various mixed cell cultures, such as primary cortical culture and hippocampal culture.

Research progress in characterizing multiple-target mechanisms of novel anti-Alzheimer's dimers / 中国药理学通报

Alzheimer's disease ( AD ) is a complex disease caused by environmental and genetic factors. Therefore, one-drug-multiple-target compounds represent the most promising pharmacological approaches to preventing and treating this dis-ease. We have previously designed and synthesized bis( n)-Cog-nitin, novel anti-Alzheimer's dimers derived from tacrine. Bis ( n)-Cognitin have been proven to act on multiple important AD targets, including acetylcholinesterase, β-secretase, N-methyl-D-aspartic acid receptor and neuronal nitric oxide synthase, con-currently. Moreover, Bis(n)-Cognitin could inhibit β-amyloid-induced neurotoxicity, decrease glutamate-induced excitotoxici-ty, reduce oxidative stress, improve learning and memory, and protect against neuronal apoptosis in various in vitro and in vivo models, suggesting that bis ( n )-Cognitin are potential anti-AD drug candidates.

Prospect of studies on four natures of ginseng,red ginseng,and American ginseng based on expression of apoptosis / 中草药

To study on the four natures (cold, hot, warm, and cool) of Chinese materia medica (CMM) is a central and knotty issue in the field of traditional Chinese medicine (TCM) and no substantial progress has been made so far, which is an obstacle in the course for the modernization of TCM. By application of apoptosis for researching the four natures of CMM, the relationship between the biological efficiency and apoptosis has been explained in a qualitative and quantitative way. A pattern recognition system for the four natures of CMM has been set up in a molecular level. That is a breakthrough to measure the four natures of CMM. Based on the abundant literatures at home and abroad, and the study done as well, the original thinking and fresh viewpoint on the orientation and development for the four natures of CMM have been put forward and then the explanation and prospective analysis on them have been carried out.