Preliminary trial of identification of brain structures in nude mice with green fluorescent protein expression by fluoroscope / 中华神经医学杂志

Objective To explore the feasibility and advantage of fluoroscope in identification of brain structures in nude mice with green fluorescent protein (GFP) expression. Methods We laid the whole brain separated from 8-week adult nude mice with GFP expression into SLY mouse brain blocker to produce slices of 1 or 0.9 mm thickness; and then,25 μm-thickness frozen sections were cut.Fluoroscope was employed to observe the morphological structure to define their anatomic structures with reference to The Mouse Brain in Stereotaxic Coordinates compiled by Paxinos. After the observation,these frozen sections were performed Nissi staining for contrast. Results Different structures can be identified by their distinct fluorescence intensity:the dense areas of nuclei,Nissl bodies and nerve tract showed low fluorescence intensity; while the structures around the areas of nuclei and nerve tract,such as,the plexiform layer of olfactory bulb and the molecular layer of cerebella,showed high fluorescence intensity.The fluorescence intensity was attenuated obviously after Nissl staining; the visualized structural information observed under stereomicroscope was in accordance with that viewed by fluoroscope.Conclusion The identification of brain structure in nude mice with GFP by fluoroscope can serve as an experimental platform being applied in the anatomic structure positioning in fluorescence tracer experiments.

Expression of SV40Tag, Rb and IRS-1 in glioma detected by tissue microarray and their relation with tumorigenesis and progression of gliomas / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To determine the expression of SV40Tag, Rb and IRS-1 in gliomas and to identify their function in gliomagenesis and progression.</p><p><b>METHODS</b>Tissue microarrays were constructed containing 118 samples including human glioma and meningioma, experimental glioma, and normal human brain tissue. The expression of SV40Tag, Rb, IRS-1, SV40Tag combined with Rb, and SV40Tag combined with IRS-1 were assayed by immunofluorescence or immunohistochemical techniques. The expression ratio and level were analyzed.</p><p><b>RESULTS</b>The expressions of SV40Tag, Rb and IRS-1 were detected in gliomas and benign brain tumors. Their positive expression rate in glioma was 65.9%, 64.6% and 48.8%, respectively, with a statistically non-significant difference between the malignant and benign brain tumors. The malignant degree was positively correlated with SV40Tag and IRS-1, but negatively correlated with Rb expression. The combined expression rate of SV40Tag and Rb was 51.2%, and the combined expression rate of SV40Tag and IRS-1 was 40.2%. In the normal human brain tissue only the expression of Rb (77.8%, 7/9) and IRS-1 (22.2%, 2/9) were detected, but expression of SV40Tag could not be observed.</p><p><b>CONCLUSION</b>Our findings that no expression of SV40Tag was observed in normal human brain tissue indicates that expression of SV40Tag may play an important role in the pathogenesis of glioma. It may be assumed that after SV40 virus invading human body, Rb disfunction and IRS-1 activation promote the malignant transformation of cells, which could be one of important factors in pathogenesis and procession of glioms.</p>

Establishment of the coculture systems of rabbit aortic endothelial cells and smooth muscle cells / 中国应用生理学杂志

<p><b>AIM</b>In order to establish a coculture system of ECs and SMCs and by which further study can be done.</p><p><b>METHODS</b>ECs in primary culture were grown on a side of Transwell membrane, and SMCs were grown on an other side of it or the bottom of culture well, so that two kinds of coculture systems were established, and detail observation on the coculture systems was carried out by transmission and scanning electron microscope.</p><p><b>RESULTS</b>ECs in primary culture were positive of VI factor by immunocytochemistry staining. ECs and SMCs were grown well on both sides of Transwell membrane, relative to ECs monolayer of "cobblestone appearance", SMCs were multilayer of "hills and valleys appearance". ECs and SMCs on both sides of Transwell membrane could form the gap junctions by micropores.</p><p><b>CONCLUSION</b>The coculture systems of ECs and SMCs were established successfully by modeling the structural relationship of vascular wall.</p>