ABSTRACT
Objective To establish an experimental subarachnoid hemorrhage(SAH)model with endo- cerebrovascular peroration.Method The right external carotid artery of SD rats were isolated,leaving a stump of approximately 3 to 4 mm.A-3-O monofilament nylon suture was inserted up through the stump of external carotid artery to the internal carotid artery for about 18～19 mm.A small resistance was usually felt,and the suture was then advanced 2 mm further and the suture was immediately withdrawn.Two hours or two days after SAH induction,SAH extension was observed.Two days after SAH induction,the diameter of the basilar artery was measured.Results SAH extends from the ipsilateral artery to the eontralateral artery after SAH induction.The diameters of basilar arteries in SAH animals were smaller than those of control rats,indicating the present of cerebrovascular spasm in SAH animals.Conclusions The endo-cerebrovascular perforation technique for establishing a non-craniotomy SAH model is reliable.
ABSTRACT
<p><b>OBJECTIVE</b>This study is aimed at developing a simple and easy way to generate dendritic cells (DCs) from human peripheral blood monocytes (PBMCs) in vitro.</p><p><b>METHODS</b>PBMCs were isolated directly from white blood cell rather than whole blood and purified by patching methods (collecting the attached cell and removing the suspension cell). DCs were then generated by culturing PBMCs for six days with 30 ng/ml recombinant human granulocyte-macrophage stimulating factor (rhGM-CSF) and 20 ng/ml recombinant human interleukin-4 (rhIL-4) in vitro. On the sixth day, TNF-alpha (TNFalpha) 30 ng/ml was added into some DC cultures, which were then incubated for two additional days. The morphology was monitored by light microscopy and transmission electronic microscopy, and the phenotypes were determined by flow cytometry. Autologous mixed leukocyte reactions (MLR) were used to characterize DC function after TNFalpha or lipopolysaccharide (LPS) stimulations for 24 h.</p><p><b>RESULTS</b>After six days of culture, the monocytes developed significant dendritic morphology and a portion of cells expressed CD1a, CD80 and CD86, features of DCs. TNFalpha treatment induced DCs maturation and up-regulation of CD80, CD86 and CD83. Autologous MLR demonstrated that these DCs possess potent T-cell stimulatory capacity.</p><p><b>CONCLUSION</b>This study developed a simple and easy way to generate DCs from PBMCs exposed to rhGM-CSF and rhIL-4. The DCs produced by this method acquired morphologic and antigenic characteristics of DCs.</p>