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 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 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 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 The retrograde isolated hepatic perfusion (RIHP) model was used to compare with the isolated hepatic perfusion (IHP) model in reducing the rate of normal hepatic tissue toxicity and peripheral drug leakage during chemotherapy in rat liver. Methods A total of 90 male Sprague-Dawley rats weighing 300-350 g were randomized into 3 groups with 30 rats in each. Group A: perfusion with Lactated Ringer'S Solution through arteria hepatica (RA) and portal vein (PV),the inferior vena cava was used as an outflow tract of perfusate. Group B: For isolated hepatic perfusion (IHP), Fluorouracil (5-FU) was added into the perfusate at a dose of 350mg/kg and introduced in to the liver through arteria hepatica, portal vein was perfused by Lactated Ringer'S Solution, and the inferior vena cava was used as an outflow tract of perfusate. Group C: by using retrograde isolated hepatic perfusion (RIHP), the solution which contains 350 mg/kg Fluorouracil (5-FU) was also introduced through arteria hepatica, the inferior vena cava was introduced with Lactated Ringer'S Solution;the portal vein was used as an outflow tract of the perfusate. On day 1, 3, 5 and 7 after the perfusion in all groups, blood serum ALT test and liver histopathology test were performed. The peripheral blood drug levels were measured with high performance liquid chromatographic(HPLC) system in group B and group C. Results The survival rate was 90%, 86.7% and 90% in group A, B and C,respectively. No statistically significant difference was observed in the survival rate among the 3groups. In all the three groups, serum ALT levels were the highest on the first day after IHP: (481.6±207.6)μmol/LingroupA;(1641. 6±658.0) μmol/LingroupBand( 913. 0±353. 5)μmol/Lin group C. Significant higher serum ALT levels were observed by comparing group B and C with A(P＜0. 05). Meanwhile, the serum ALT levels were significantly higher in group B than in group C (P＜0.05). The peaks of peripheral blood drug concentration during the perfusion were 131.2±29.4μg/ml in group B and 65.3±28. 4μg/ml in group C. Significant difference was observed (P＜0. 05). Liver biopsies of group A showed mild changes on the first day after IHP and returned to normal after 7 days. Group B showed severe changes on the first day after IHP and local necrosis still existed after 7 days. Group C showed moderate changes as compared with group B on the first day after IHP and also returned to normal after 7 days. Conclusion Retrograde isolated hepatic perfusion (RIHP) can reduce the liver toxicity compared to isolated hepatic perfusion (IHP). Hopefully, RIHP will be considered as a safer way in regional chemotherapy in liver cancer.

Objective To study the clinical outcomes of 28 pancreatic cancer patients who underwent total pancreaticoduodenectomy.Method The clinical data of 28 patients with pancreatic cancer who underwent total pancreaticoduodenectomy from January 2009 to March 2015 were retrospectively analyzed.Results Among the 28 patients,complications occurred in 11 (39.2％) after the operation.There were 7 patient having Grade Ⅱ,4 Grade] complications.No patient died within 30 days after the operation.Fol low-up of 25 patients showed a median survival of 13.5 months.There were 24 patients with pancreatic ductal adenocarcinoma,and the median survival was 13 months.Conclusions Total pancreaticoduodenectomy could not improve long-term survival but it decreased postoperative complications and improved postoperative quality of life.In selected patients,total pancreaticoduodenectomy could be a rational option.

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 study the transfection efficiency, protein expression, and effect of adenovirus-mediated transfection on microvascular endothelial cells transfected by angiopoietin-related protein 2(Ad. ARP2)gene. Methods Mice coronary microvascular endothelial cells(CMECs) were isolated, cultured and transferred by Ad-ARP2. The transfection efficiency and cellular toxicity of adenovirus vector to CMECs were detected by immunofluorescence staining. Expression of Ad. ARP2 in CMECs and the secreted materials in culture medium were measured by Western blot and ELISA and compared among groups of Ad. ARP2, Ad. null, and PBS control. Vascular endothelial cells were incubated with conditional medium containing secreted ARP2, and effects on cells sprouting were observed in matrigel. Results Adenovirus-transfected CMECs showed a very high efficiency. When multiplicity of infection (MOD was 200, the transfection efficiency was 93. 5% ,and no harmful effect on CMECs growth was found. When CMECs were transfected with Ad. ARP2, there was a high ARP2 expression, which was significantly different from that with Ad. Null or PBS. The conditional medium containing ARP2 had an excellent ability to stimulate sprout of CMECs which phenomenon could not be seen in the control groups. Conclusions Adenovirus vector can be transferred into CMECs efficiently and safely. Ad. ARP2 gene transfection allows a high transient expression, and the expression products can stimulate the sprout of microvascular endothelial cells in vitro very well.