BACKGROUND: Both density gradient centrifugation and adherence method arc frequently used to isolate bone marrow mesenchymal stem cells (BMSCs).OBJECTIVE: To investigate the approaches to isolate, culture and identify the rabbit BMSCs in vitro by the combination of den,ity gradient centrifugation and adherence method. DESIGN, TIME AND SETTING: Contrast cytological study, which was performed in Central Laboratory of Shanghai 6th People's Hospital between October 2007 and March 2008.MATERIALS: Six 2-week-old rabbits were selected for BMSCs preparation and primary culture; Percoll separating medium (1.073 kg/L) was also used for this study.METHODS: BMSCs were separated and purified with Percoll separating medium by density gradient centrifugation and adherence method. The three-, five-, seven-, and nine-passage BMSCs were counted for growth curve. MAIN OUTCOME MEASURES: Morphological features and growth states of primary and passage cells were observed under inverted microscope. Indirect immunofluorescence of CD44 and CD34 antibodies were used to examine the stem cells. CD44 staining was positive, and CD34 staining was negative, suggesting the extracting and purifying cells were BMSCs. RESULTS: The passage BMSCs were uniformly distributed like fusiform shape, which were more uniform than primary cultured cells. The BMSCs grew productively and proliferated rapidly; meanwhile, the nucleolus was clear, caryopla.sm was in a large proportion, morphological features were uniform, ceils like bostrychoid or whirlpool were arranged parallelly, and the five-pa.ssage cells were not changed remarkably. Proliferation was decreased gradually with the passage increasing; especially, the proliferation of three-five-passage cells was the strongest. The separated cells expressed CD44 but not CD34. CONCLUSION: High-purified rabbit BMSCs are obtained by both density gradient centrifugation and adherence method.

Objective:To observe cellular,humoral and mucosal immune responses induced by DNA-or protein-based of FimH of UPEC type 1.Methods:After mice were immunized respectively with recombinant plasmid pcDNA3.1/fimH or pcDNA3.1/fimC,and the combinant FimH and FimC protein,the anti-FimH protein IgG of sera and SIgA in bladders were detected by ELISA.The lymphocyte phenotypes of CD3,CD4 and CD8 were analyzed by FCM.Results:The titers of IgG in sera and SIgA in the bladders were all low in the group immunized by recombinant FimH plamid,but the percentage of CD4+T cells in spleen was high,which revealed that recombinant FimH plamid was able to trigger better cellular immune response.The titers of IgG were very high in the group immunized by FimH protein,which suggested that the FimH protein was able to trigger better humoral immune response,but SIgA in the bladders was not detectable.Conclusion:The DNA for FimH can induce humoral,mucosal and cellular immune response.FimH protein can only induce humoral immune response.FimC protein is able to enhance the immunogenicity of FimH protein.

Objective To explore the value of improved CT perfusion in delineation of brain arteriovenous malformation(AVM) target for stereotactic radiosurgery.Methods 22 patients diagnosed with AVM by DSA were included in this study.14 cases of AVM were detected from initial symptoms of intracereb.ral hemorrhage,of which 4 cases were given immediate intracranial hematoma evacuation,then in 3 cases postoperative embolization was performed,and other 10 cases received conservative treatment including 3 cases treated by embolization.8 cases of AVM were detected from initial symptoms of epilepsy or headache,without surgery or embolization treatment.In all patients,the improved CT perfusion and MRA examinations were performed before treatment to evaluate the diagnostic efficacy of different methods in AVM.Results The interference rates of hemorrhage and granulation tissue on MRA images were 27.3 ％ and 54.5 ％,respectively,while those on enhanced CT and improved CT perfusion images were 0 ％.The interference rate of embolization material on enhanced CT and improved CT perfusion images was 27.3％,while that on MRA images were 0％.The contrast-enhancement rates of MRA,CT and improved CT perfusion images were 4 5.5 ％,5 4.5 ％ and 7 2.7 ％,respectively.Conclusion Improved CT perfusion technique is helpful in delineation of brain AVM target for stereotactic radiosurgery in patients with AVM combined with intracerebral hemorrhage or postoperative patients.