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.

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

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

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

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 explore the effect of adenovirus-mediated bcl-2 transfer on the regeneration and repair of peripheral nerves in rats. Methods The sciatic nerve-transected model was selected and sutured end to end with epineurium. Then, adenovirus/sense-B-lymphoma/leukemia-2 (Ad/s-bcl-2), adenovirus/anti-sense-B-lymphoma/leukemia-2 (Ad/as-bcl-2), adenovirus/?-galactosidase (Ad/LacZ) and normal saline (NS) were injected into the sciatic nerve 0.5 cm away from the sutured point respectively. The rats were perfused with 4% paraformaldehyde at the 48th hour, 7th, 15th and 30th days respectively after operation. The spinal cords L 4-6 were harvested, frozen sectioned and given x-gal staining, bcl-2 in situ hybrid and immunohistochemical staining and GAP-43 immunohistochemical staining. The postoperative functional recovery of the rear limbs of rats was dynamically observed by using sciatic nerve functional index (SFI). Results The GAP-43 expression in the motor neurons of anterior horn of spinal cord in group Ad/s-bcl-2 was higher than that in other groups (P0.05). Conclusions Gene transfer of bcl-2 can promote regeneration and functional recovery of the peripheral nerves following sciatic nerve injury.

Objective To explore the spatio-temporal regularity of cerebral edema in the early stage of severe burn (50% TBSA Grade Ⅲ) with four-dimensional (4D) mathematic model. Methods Twenty-six dogs were randomly divided into control group and 6, 12, 18 and 24 hours post-burn groups. The dogs in the postburn groups suffered severe burn of 50% TBSA GradeⅢ. Six hours after burn, 50 g/L glucose was injected according to Parkland formula. MRI manifestation, histopathological changes, brain water content and intracranial pressure were observed in each group. A 4D mathematic model was established based on the results of MRI scanning. Results Two turning points ( 6 and 18 hours postburn) and three phases of pathologic changes were displayed by 4D mathematic model of cerebral edema in the early stage of severe burn. The first phase (6 hours postburn) was in the subclinical period,when an effective treatment should be taken as quickly as possible so as to prevent deterioration of postburn cerebral edema. The second phase ( 6-18 hours postburn) was in the apostatic period with pathologic characteristic of vasogenic and cytotoxic brain edema. The third stage (18-24 hours postburn) was the critical period of cerebral edema. Intracerebral pressure (ICP) increased rapidly due to limitation of cranial cavity, when cerebral hernia easily happened. Conclusions The 4D model indicates that there is a feature of type S curve in the pathologic process of cerebral edema in the early stage after severe burn.