OBJECTIVE To study the biocompatibility of skin reproductive membrane. METHODS According to ISO’s standards, the extractions of the skin reproductive membrane were prepared, and the acute systematic toxicity test, primary skin irritant test, cytotoxicity test, gene expression of type I collagen and fibronectin were detected to evaluate the biocompatibility of skin reproductive membrane. RESULTS All of those tests showed negative results. CONCLUSION The skin reproductive membrane has excellent biocompatibility in the level of the systematic, cellular and molecular biology.
Objective To observe the altered gene expression of ECV304 cell after treated with endostatin (ES) using cDNA microarray, and to investigate its molecular mechanism.Methods BiostarH40S gene expression DNA microarrays were used to profile changes in gene expression of ECV304 cells after treated with ES for 36 hours. To prepare the probes, mRNA from both control and treated cells were isolated and purified, and then reversely transcribed to cDNA with the incorporation of fluorescentlabeled dUTP: Cy3 and Cy5 respectively. The probes were hybridized with expressed cDNA microarray, the fluorescent signals of Cy3 and Cy5 were scanned and analyzed by a computer system.Results ES inhibited the proliferation of ECV304 cells. The result of electrophoresis indicated that the extracted total RNA had high quality.Approximately 5.96% of all human genes examined on 4096 gene microarrays showed altered expression, with 225 downregulated genes and 19 upregulated genes.Conclusion The expression of some genes of ECV304 cells decrease after treated with ES, which may be related to the mechanism of inhibition of neovascularization.
Objective To investigate the expression of T cell receptor (TCR) Vβ8.3 gene on CD4+ T lymphocytes in the rats with experimental autoimmune uveoretinitis (EAU). Methods Eighteen Lewis rats were divided into EAU, complete Freund′s adjuvant, and the control group. Inter photoreceptor retinoid-binding protein (IRBP) R16 peptide was synthesized using Fmoc procedure for induction of EAU. Magnetic absorption cell sorting (MACS) me thod was used to isolate the CD4+T lymphocytes from the spleen of the rats. Flow cytometry was used to monitor the efficiency of isolation. The expression of TCR Vβ8.3 gene segment on CD4+T lymphocytes was determined by fluorescent quantitative polymerase chain reaction. Results EAU was successfully induced in the Lewis rats immunized with IRBP R16 peptide. The proportion of CD4+T lymphocytes isolated by means of MACS was statistically higher than that before isolation (P＜0.001). The expression of TCR Vβ8.3 gene segment on CD4+ T lymphocytes in EAU rats was significantly higher than that in the control (P＜0.05). Conclusions There is a predominant usage of antigen-specific TCR Vβ 8.3 gene in EAU rats induced by IR BP R16 peptide, which may serve as a target for immunotherapy of EAU. (Chin J Ocul Fundus Dis，2004，20：165-167)
Objectives To investigate the expression of pax-6 in ret ina of in fant monkeys with myopia induced by optical defocus, and to determine the role of pax-6 would play or not in onset and development of myopia and emmetropization.Methods Nine healthy infant rhesus monkeys, aged from 1 to 3 months, were selected and wore spectacle lenses or underwent photorefractive keratectomy (PRK).Transcription polymerase chain reaction method and quantitative analysis were used to determine the expression of pax-6 in the retina with myopia induced by optical defocus in different time, and the result was compared with that in retina without myopia.Results The myopia caused by hyperopic defocus was found. The expression of pax-6 in the retina with myopia induced by optical defocus was significantly higher than that in the retina without myopia（t=3.480，P=0.004）.Conclusions The expression of pax-6 is enhanced by hyperopic defocus in the infant monkey retina, which suggests that pax-6 may be involved in vision-dependent eye growth and emmetropization. (Chin J Ocul Fundus Dis，2003，19：201-268)
OBJECTIVE: To construct eukaryotic expression plasmid of mouse myogenic regulatory factor MyoD gene for further study on MyoD gene function in molecular regulatory mechanism in skeletal muscle repair. METHODS: The plasmids PEMMBC2 beta 5 containing full cDNA length of MyoD inserted in EcoRI restriction site, were first propagated in Escherichia coli DH5a, then extracted and purified with the Wizard Plus Minipreps DNA Purification System (Promega, USA). The coding sequence of MyoD in PEMMBC2 beta 5 was confirmed by agarose gel electrophoresis and DNA sequence analysis. After plasmids PEMMBC2 beta 5 and plasmids pcDNA3-neo were prepared by digestion with EcoRI, the MyoD cDNA fragment was inserted into EcoRI site in pcDNA3-neo eukaryotic expression vector, and pcDNA3/MyoD was formed. The pcDNA3/MyoD, digested with restriction enzymes, was found to contain the MyoD cDNA sequence by agarose gel electrophoresis analysis. RESULTS: The extracted and purified PEMMBC2 beta 5 contained the correct nucleotide sequence for the full length of MyoD cDNA fragment. The MyoD cDNA fragment had been inserted into the eukaryotic expression plasmid pcDNA3-neo, which formed the pcDNA3/MyoD. CONCLUSION: The pcDNA3/MyoD, a eukaryotic expression plasmid, for MyoD is constructed successfully.
Objective To review the advances in gene expression of marrow stromal cells(MSCs) in biological characteristic, differentiation, gene therapy, supporting hematopoiesis, inflammation reaction in wound repair.Methods The related articles in recent years were extensively reviewed.Results MSCs can express not only specific mRNA of mesodermal cells but also that of endodermal and ectodermal cell types on various differentiation conditions. When transfected or transducted, MSCs can steadily express many therapeutic transgenesin vitro or in vivo as well. Furthermore, they have the ability to support hematopoietic system and participate in the process of inflammation in wound repair.Conclusion MSCs are stem cells which have the characteristicof self renewing, multipotency and easy to expand in vitro. MSCs are ideal target cells for cell and gene therapies.
Purpose To investigate the expression of intercellular adhesion molecules ICAM-1 and Mac-1，in epiretinal membanes (ERM) of eyes wi th proliferative vitreoretinopathy (PVR)． Methods Twenty epiretinal membranes were obtained from eyes undergone vitrectomy for retinal detachment complicated with PVR and observed by immunohistochemical examination． Results Expressions of ICMA-1 and Mac-1 were observed in 18 and 15 membranes respectively．Expression of both adhesion molecules in 12 membranes． Conclusion The findings indicate that adhesion molecules might be involved in the development of PVR． (Chin J Ocul Fundus Dis，2000，16：71-138)