ABSTRACT
Objective:To construct an adenoviral vect or carrying human tissue inhibitor of metalloproteinase-2(TIMP-2)gene in order to mediate the expression of TIMP-2 gene in vascular smooth muscle cells(SMC)i n vitro. Methods:A recombinant adenovirus(AdhTIMP-2)containging human TIM P-2 cDNA fragment was generated by homologous recombination in BJ5183 bacteria. Recombinant plasmids were screened by alteration of antibiotics.The adenovirus v ector was then packaged and amplified in 293 cells.The SMC of rat aortic were is olated and cultured in vitro and been infected with AdhTIMP-2.The expression of TIMP-2 was detected by the techniques of Western blot and RT-PCR. Results:The recombinant adenoviral vector carrying human TI MP-2 was constructed. The titer was 4?1011efu/ml after purification.The AdhTIMP-2 could infect the cultured VSMC efficiently(MOI=100,infection ra te=94%).The expression of TIMP-2 gene in those infected cells was detected by R T-PCR.After the cells were infected with AdhTIMP-2 24hours,TIMP-2 protein cou ld be detected in the conditioned medium by Western blot. Conclusion:The recombinant adenoviral vector carrying human TIMP -2 is successfully constructed and AdhTIMP-2 can efficently mediate the expres sion of TIMP-2 gene in cultured VSMC,paving the way for further application in vascular disease gene therapy.
ABSTRACT
Objective To investigate the effect of TIMP-2 gene that was transfected by adenovirus on extracellular matrix of abdominal aortic through assessing the changes of morphology and histopathology of the rat models with abdominal aortic aneurysm. Methods The rat models with abdominal aortic aneurysm were constructed by intraluminally perfusing porcine pancreatic elastase. Twenty-four SD rats with aneurysm were then randomly divided into 3 groups: AdTIMP-2 group (perfused locally with solution of TIMP-2 gene transfected by adenovirus vector to abdominal aorta), AdCMV group (transfected by non-viral vector), and PBS group. After 14 days, the concentrations of elastin and collagen that were collected from the samples of aortic wall were measured by image analysis system and the fixed aortic tissues were examined by light microscopy and some other specific staining methods. Results None of abdominal aortic aneurysm developed in TIMP-2 gene transfected group, with significantly higher rates of developed aneurysm in the other groups (P
ABSTRACT
ObjectiveTo discuss the epidemiological characteristic s of road traffic injuries in the Qinghai-Tibet Plateau and explore effective pre ventive measures against traffic accidents in the plateau as well as correspondi ng emergency treatment methods.MethodsA retrospective statis tical analysis was carried out in 1 894 cases of traffic injuries with detailed data admitted in our hospital from January 1980 to December 2000. Resu ltsMost of the traffic accidents occurred on the Qinghai-Tibet road, accounting for 68.0% (1 288/1 894). Of the patients with serious traffic inju ries, 54.0% (1 022/1 894) were in need of hospitalization. The patients who first entered into the plateau accounted for 61.1% (1 158/ 1 894). Death occurre d in 108 cases accounting for 5.7%. The duration from injury to treatment was ve ry long, average over six hours. Furthermore, the patients received no any manag ement before treatment in the hospital. ConclusionsHypoxia i n the plateau, unclear road sign and fatigue driving are the main factors leadin g to traffic accidents. Lack of health care units, serious hypoxia, long deliver y time and carriers' deficiency in medical knowledge are the main causes for gr eat amount of casualties.