Objective:To evaluate the protective effects of adenovirus-mediated gene transfer of soluble complement receptor type 1(sCR1) on acute myocardium ischemia in mice.Methods: Twenty-seven SD rats were subjected to left anterior descending coronary artery(LAD) occlusion(30 min) and reperfusion.In the treatment group(n=14),a mixture of adenovirus carrying sCR1 and LacZ was injected into the ischemic zone(100 ?l,10~(10)pfu)5 min before reperfusion;in the control group,only adenovirus carrying LacZ was injected(n=13).Echocardiography was performed 2 weeks later,followed immediately by pathologic examination.Results: Echocardiographic results of the treatment group were better than those of the control group((P

Objective To culture normal adult esophageal epithelial cells and establish a normal esophageal epithelial cell line to provide experimental materials for further studies in vitro.Methods Normal esophageal epithelium samples were obtained in sterile phosphate buffered saline supplemented with antibiotics from the normal part of the esophagus of a patient with esophageal carcinoma.Tissue was cleaned of all connective tissues and muscles and rinsed in phosphate buffered saline 5-6 times.The mucosal layer was minced into small pieces and dissociated into a single cell suspension by 0.25% Dispase and 0.25% trypsin/0.02%EDTA.Cells were grown in keratinocyte serum-free media.The cultured cells were identified through their morphological characteristics and immunohistochemical staining.The proliferative capacity of the cultured cells was also examined.Results The cultured cells showed microscopic features of epithelial cells and were positive in keratin and epithelial membrane antigen staining.Eight days after primary culture,the cells displayed a cobblestone morphology reaching 80%-90% confluency and were passaged successfully with 0.25% trypsin/0.02%EDTA.The cell cycle analysis showed about 77.60% of the cultured cells was in G0/G1 phase and the others in S/G2/M phase.Conclusion\ The culture methods and techniques used in the experiments are convenient and suitable for the primary culture and subculture of normal adult esophageal epithelial cells.

Gap junction, a special membrane structure communicating many kinds of cells, is composed of connexin.Changes of connexin levels and distribution can lead to changes of conduction velocity of electric couple and increase of reentrant arrhythmia, finally to atrial fibrillation. This article elucidates the relationship between cardiac gap junction protein connexin and atrial fibrillation.

Objective To construct the replication-deficient recombinant adenovirus coding ARP2 gene before the studies of gene transfection to ischemic myocardium.Methods From Dec.2004 to Oct.2005,in the Department of Cardiology,Changhai Hospital of the Second Military Medical Univesity,the ARP2-pAxCAwt was constructed by inserting the cDNA of interest into the SwaI site of pAxCAwt.Results The right direction of the insert was confirmed by restriction.Conclusion The direction of the insert must be confirmed by restriction analysis,and the DNA package protein is much helpful during the transfection of the recombinant cosmid to E.coli.

Objective: To investigate the angiogenic potentiality of angiogenin in ischemic myocardium and estimate the possibility of its application in biologic treatment. Methods: Recombinant human angiogenin derivative Asp116His was applied in an acute myocardial infarction model in rabbits which were established by direct intramyocardial injection into the border zone of the ischemic myocardium. The animals were sacrificed on the eighth postoperative day and their hearts were harvested and subjected to histologic studies. Results: Myocardium around the injection sites was apparently less ischemia in the experiment group as compared with control groups. A large number of small vessels were observed in the interstitial tissues and even more were revealed in the myocardium on immunohistochemistry studies. Conclusion: The results indicate for the first time that angiogenin or its derivatives may act on ischemic myocardium and stimulate neo angiogenesis, which holds great promise for their future application in biologic revascularization of ischemic myocardium.

Objective To observe the inhibition of vascular endotheilal growth factor (VEGF) gene transcribition and translation with antisense VEGF165 adenovirus vector transfection. Method To transfect the synovial cells with antisense VEGF165 adenovirus and detect the gene translation level with northern-blot method. Result The Northern-blot showed that: the transcription of VEGF gene were inhibited markedly after the antisense VEGF165 gene transfection for 3 days and the inhibition efficiency was obivious in 4th day. In the 5th day, the VEGF translation signal was hardly detected. Conclusion The antisense VEGF165 adenovirus gene transfection can inhibite not only the transcription but the translation of VEGF gene in synovial cells. Thus it can inhibit the secretion of VEGF in synovial cells with high efficiency.

Objective To discuss the harm and the rectification procedures for patients who have received nonstandard cholangioenterostomy and improper endoscopic retrograde biliary drainage.Method The clinical data of 55 patients who had received nonstandard cholangioenterostomy and improper endoscopic retrograde biliary drainage seen at the Ningbo LiHuiLi Hospital between 2004.6 to2011.12 were retrospectively analyzed.Results There were 23 patients who had stones located intrahepatically which had not been dealt with in the previous operation; 16 patients received choledochoduodenostomy; 5 patients received side-to-side or side-to-end cholangiojejunostomy (with no division of the common bile duct) ; 2 patients developed anastomotic stricture after nonstandard cholangioenterostomy; 3 patients had a short efferent loop of jejunum; and 6 patients had improper endoscopic retrograde biliary drainage.Through reoperative rectification,all patients had satisfactory therapeutic outcomes.Conclusions For hepatolithiasis patients,nonstandard cholangioenterostomy and improper endoscopic retrograde biliary drainage not only cause harm to the patients,but also force patients to have a reoperation.Thus,following strict operative indications,choosing the right operation and improving on the operative skills are the keys to prevent a reoperation.

Objective:To modify the pulsatile bioreactor we constructed previously for simulating the high-flow, high-pressure hemodynamics of heart valve in vivo, and to experimentally cultivate the tissue-engineered heart valves (TEHV) in the modified bioreactor. Methods: T-PLS system (NewheartBio Co., Ltd Korea) was used to generate pulsatile flow in the modified bioreactor and we designed a new air-exchange pathway to avoid contamination. The TEHV were made by seeding human bone mesenchymal stem cells (BMSCs) on de-cellularized porcine heart valve. After cultured under static condition for 4 d, the TEHVs were moved to the modified bioreactor and exposed to low-flow (0-600 ml/min) or high-flow(0-4 800 ml/min) pulsatile hydrodynamics for 7d, then the cells on TEHVs were evaluated. Results: After modification, the flow range expanded from (0-1 200) ml/min to (0-6 000) ml/min and the pressure range expanded from (0-40) mmHg to (0-180) mmHg. In culture experiments, 26.3% of the seeded cells remained under low-flow environment and cells were completely lost under the high-flow dynamics. Conclusion: The modified bioreactor can basically simulate the dynamics of heart valve in vivo and can be used in TEHV cultivation research. However, the current TEHV can not tolerate the high-flow pulsatile hydrodynamics.

Objective:To investigate the survival of rabbits receiving autologous left auricle cardiomyocytes transplantation into the infarcted myocardium and to assess the effect of transplantation on the peripheral blood supply, heart rate, and cardiac function. Methods: Healthy adult rabbits were randomly divided into transplantation group (n=8) and control group (n=8). The myocardial infarction (MI) models were established by ligating the left anterior descending arteries in all rabbits. Four weeks later the left auricles were harvested and the auricle cardiomyocytes were isolated and labelled with DAPI ex vivo. Rabbits in the transplantation group were injected with DAPI-labelled cell suspension into the infarcted areas and those in control group received culture medium. All the rabbits were examined by electrocardiogram (ECG) and 2-D ultrasonic cardiogram (UCG) 4 weeks after transplantation. Specimens were harvested from the transplanted areas and observed histologically. Results: All rabbits survived 4 weeks after transplantation. ECG showed that the heart rate in transplantation group was faster than that in control group (P

Vascular endothelial growth factor (VEGF)165 cDNA were inserted into the expression plasmid pcDNA3.1 site in an antisense orientation to produce antise VEGF165 expression vector.Then electrophoresis and sequencing were carried out,the antisense vector were verified and could be used for related study of antisense gene expression.