Objective To observe the effectiveness of transferring human A20 gene into endothelial cells. Methods The shuttle plasmid pCAGGSEHA20 was constructed using gene cloning and recombined technique. Endothelial cells were transfected with pCAGGSEHA20 and pMAMneo by DOTAP. The postive cell clones were selected with G418. The stable transfection and expression of A20 in the endothelial cells were determined by in situ hybridization and immunohistochemical analysis. Results The two fragments digested from pCAGGSEHA20 by EcoRⅠ represented 4 6?kb and 2 3?kb by electrophoresis, which were confirmed to be the carrier and the A20 gene fragments inserted originally. The above results indicate that the construction of pCAGGSEHA20 was successful. Abundant A20 stable expression in endothelial cells transfected with pCAGGSEHA20 was confirmed by in situ hybridization and immunohistochemical analysis.Conclusion By means of the DOTAP, hA20 gene can be transferred and stably expressed in endothelial cells.

The origin of 200 occipital arteries on 100 cadavers were investigated. 199 cases of the occipital arteries originated from the external carotid artery and only one was found arising from the common carotid. The artery arised as an independent stem in 80.5% of cases and from a common stem with other arteries in 19.5%. In 85.6% of cases the site of origin of the artery was within 10mm above or below the mandibular level. In addition, the sites of origin of the occipital and facial arteries were compared. It was found that the site of the occipital artery was usually higher than that of the facial artery in contrary to the statement of the conventional textbooks that they are in a level opposite to each other.

Objective To construct replication deficient adenovirus vector AdCMV lacZ Methods Recombinant adenovirus shuttle plasmid pAdCMVlacZ was first constructed and then lacZ cDNA cloned into Hind Ⅲ and Eco R Ⅴ of adenovirus shuttle plasmid pAdCMV by conventional methods Results The restriction enzymatic map of pAdCMVlacZ accorded with theoretic analyses and pAdCMVlacZ showed blue bacterium groups under the existence of revulsant and X gal Conclusion The adenovirus shuttle plasmid pAdCMVlacZ has been constructed successfully

The microvasculature of the rectus abdominis muscle in dogs was observed under scanning electron microscope. The specimens were treated by vascular corrosion east and enzymic digestive methods. Some new results had been obtained.The intramuscular arterioles penetrate the muscular fasciculus with almost right angle. The capillaries go along the myofibers oriented parallel to each other and arrange fascicularly. The morphologically distinct precapillary sphincters are apparent at the original region of the capillaries. There are abundant anastomosing channels between the capillaries which constitute ladder-like microvascular networks encircling the myofibers. Sometimes, the calibre was partial enlarged in the anastomosing channel, which may be beneficial to store of the blood oxygen.The physiological significance of the intramuscular microvasculature was discussed.

Objective To explore the changes of wall shear stress(WSS) effect on endothelial cell morphology remodeling after reducing arterial flow. Methods The reducing flow of common carotid artery was established in 60 rabbits,and arterial stretched preparations were made at 8 different intervals from 0 to 30 days.The shape and fibrous actin(F-actin)of arterial endothelial cell(AEC)were observed under an microscope and confocal laser scanning microscope(CLSM) using silver and fluorescein-phalloidin stain. Results The fewer stress fibers were viewed in AEC and its shape changed from spindle to ellipse after WSS decreased for 12 hours,the value of shape index(SI) and length/width(L/W) was greatly higher and lower than the levels of control,respectively(P

AIM:To explore the effect of wall shear stress(WSS) on arterial endothelial cell(AEC)proliferation. METHODS: The reducing flow of common carotid artery was established in 60 rabbits,and arterial endothelial cell(AEC)proliferation were assessed at 8 different time intervals from 0 to 30 days. RESULTS: With progressively increasing in WSS, the rate of proliferation indicated a much higher level from 7 days to 30 days than control( P

Objective　To explore the changes of ER-IR and the ultra structure in the medial preoptic area, arcuate nuclei of early-aged mice treated with estrogen. Methods　Immunohistochemistry assay and electron microscopy were used in this study. Results　ER-IR in the medical preoptic area and arcuate nuclei were greatly reduced after estrogen was given. The cell nuclei of neurons in these areas migrated towards the side, the nuclear membrane became folded, synapse became richer, and the number of synapse vesicle increased. Conclusion　Estrogen can affect the neuron structure and function through the change of estrogen receptor expression in the medial preoptic area and arcuate nuclei of the hypothalamus cardiovascular center.

Objective To investigate the effects of zinc finger protein gene A20 on the inhibition of lipopolysaccharide (LPS) induced interleukin 8 (IL 8) expression in endothelial cells. Methods Plasmid pcDNA3.1EHA20 was transfected into human umbilical vein endothelial cells (HUVECs) by DOTAP method. The positive cell clones were screened with G418. The stable transfection and expression of A20 in HUVECs were determined by immunofluorescent analysis. IL 8 expression was detected by sandwich ELISA with double monoclonal antibodies. Results High expression of A20 gene in HUVECs transfected with pCDNA3.1EHA20 was confirmed by immunofluorescent analysis. IL 8 expression increased in LPS induced endothelial cells. A20 gene could inhibit more than 70% LPS induced IL 8 expression ( P

Objective To observe the growth and differentiation of the cultured mouse bone marrow stromal cells in vitro into neural like cells. Methods Bone marrow stromal cells (BMSCs) isolated from mice were cultured according to the routine method, and induced to differentiate by basic fibroblast growth factor b (bFGF), retinoic acid (RA), nerve growth factor (NGF), and dimethylsulfoxide (DMSO). Expressions of neural microfilament protein (NF 200), glial fibrillary acidic protein (GFAP), and fibronectin at 72 h after inductin were determined by immunoflurenscence and immunocytochemicalmethod. Results At 72 h after inductin, neural like changes were found in BMSCs. Immunohistochemical stain showed that 60% BMSCs differentiated into NF 200 positive cells, and about 50% BMSCs into GFAP positive cells. Fibronectin expression was significantly lower as compared with that in the control group. Conclusion Combined action of RA, bFGF, NGF, and DMSO can induce BMSCs to differentiate into neural like cells in vitro .

Objective To acquire NF-?B p65 TAD(transcription activation domain) and construct its eukaryotic expression vector and express it in endothelial cells.Methods Human umbilical vein endothelial cells(HUVECs) were cultured and total RNA was extracted.The p65 TAD gene was amplified by RT-PCR.After sequencing,the p65 TAD gene was inserted into the eukaryotic expression vector pEGFP-N1 with the green fluorescence protein,named pEGFP-N1-p65 TAD.pEGFP-N1-p65 TAD was transfected into HUVECs and its expression was observed under fluorescence microscope and analyzed by Western blotting.Results p65 TAD(288-548) was cloned successfully.The constructed plasmid including p65 TAD gene was identical to the designed.p65 TAD gene was expressed successfully in HUVECs.Conclusion The construction of eukaryotic expression vector including p65 TAD gene and its expression in HUVECs are very successful.