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BACKGROUND: Endothelial progenitor calls are the cells that can form new blood vessels in the way of angiogenesis in the body,which updates the conventional theory of angiogenesis, vascular damage and repair after birth and provides new ideas for research and treatment of ischemic diseases.OBJECTIVE: To investigate the effects of dog umbilical cord blood endothelial progenitor call (UCB-EPC) transplantation on angiogenesis after myocardial infarction.DESIGN, TIME AND SETTING: An in vivo cytological experiment was performed at the Laboratory Center of Xinhua Hospital between May 2006 and March 2007.MATERIALS: One full-term pregnant hybrid dog was included for preparation of UCB-EPCs. Thirty-six adult dogs were randomly divided into a cell transplantation group (n = 18) and a model control group (n = 18).METHODS: Acute myocardial infarction model was established in each group by ligation of antedor descending coronary artery.In the cell transplantation group, 2 mL physiological saline containing 5×10<'6> BrdU-labeled EPCs was injected into the coronary artery, while in the model control group, simple physiological saline of the same amount was given. At 1,4, and 8 weeks after transplantation, dogs were sacrificed for harvesting myocardial tissue.MAIN OUTCOME MEASURES: Myocardial infarction was confirmed by hematoxylin-eosin staining. Myocardial angiogenesis was observed by BrdU immunohistochemical staining. The number of infarcted myocardial vessels was calculated by yon Willebrand (vW) factor staining.RESULTS: There was plenty of scar tissue, flbroblasts, and small vessels in the myocardial infarction region. In the cell transplantation group, brown yellow particles (BrdU-positive expression) appeared in some nuclei in small vessels from infarcted myocardium. Newly formed vessels were not found in the model control group. In the cell transplantation group, brown yellow particles (vW factor-positive expression) appeared in the cytoplasm of the vascular endothelial cells in the myocardial ischemia and infarction regions, vW factors were not expressed in the model control group. At 1, 4, and 8 weeks after myocardial infarction,there was no significant difference in vessel counts no matter in myocardial ischemia region or in myocardial infarction region between the call transplantation and model control groups.CONCLUSION: EPCs derived from UCB of pregnant dog can participate in the formation of blood vessels but can not promote angiogenesis after acute myocardial infarction.
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BACKGROUND: Umbilical cord blood-mesenchymal stem cells (UCB-MSCs) following differentiation into cardiomyocytes were transplanted into ischemic myocardium. The transplanted cells can build connection with host cells and repair the infarct myocardium. OBJECTIVE: To detect the cell-cell junction after transplantation of the cardiac-like cell derived from the canine umbilical cord blood stem cells. DESIGN, TIME AND SETTING: A randomized controlled animal study was performed from July 2006 to October 2007 at the Animal Experimental Center of Xinhua Hospital Affiliated to School of Medicine, Shanghai Jiao Tong University. MATERIALS: A total of 2 full-term pregnant canines were used for isolation of UCB-MSCs. A total of 36 adult mongrel canines were divided into cell transplantation group and model control group (n=18) according to the rule of random digits table. METHODS: The MSCs at passage 4 were transfected by Laz-Z. After 3-day culture, MSCs were induced by 10 μmol/L 5-azacytidine (5-aza). The canine models of myocardium infarction were established following 3 weeks of culture. 2 mL (1 ×107)MSCs were transplanted into dogs with acute myocardium infarction by coronary artery infusion and local injection in cell transplantation group. An equal volume of saline was used in the model control group. The specimens were harvested and detected at 2, 4 and 8 weeks, respectively. Cell junction was determined using immunohistochemistry. MAIN OUTCOME MEASURES: The following parameters were measured: gene trensfection, myogenic induction and differentiation results of UCB-MSCs; junction of transplanted cells and host cardiomyocytes. RESULTS: Following 72 hours of transfaction, mass of cells expressed LacZ gene, synthetized galactosidase, and stained blue using X-gal staining. Following 3 weeks of 5-aza induction, the antigen a-Actin, Desmin and Connexin43 were all been positively expressed, but before induction they were all negative. From the myocardial section of 8 weeks after transplantation, the junction was formed between the transplanted cells and the host myocardium as formed between the transplanted cells. In the junction, green-fluorescence positive expression of cadherin and connexin43 could be seen. However, in the model control group, only cadherin and connexin43 expressed positively, but the transplanted UCB-MSCs with red fluorescence could not been observed. CONCLUSION: The UCB-MSCs is able to differentiate into cardiac-like cell in vitro and form cell-cell junction in vivo to communicate with surrounding cells.
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BACKGRoUND:Endothelial basal medium is mainly used to culture endothelial progenitor cells.Studies on mesenchymal stem cells (MSCs) cultured in this medium are few. OBJECTIVE:To compare the outcome of MSCs cultured in different mediums including endothelial basal medium. DESiGN.TIME AND SETTING:The control eell experiment was performed at the Muniopal Key Laboratory of Xinhua Hospital of Shanghai.China from September 2005 to May 2006. MATERIALS:Eight pregnant mongrel dogs were selected to obtain umbilical cord blood for isolation and culture of stem cells. Endothelial eell basal medium and endothelial cell medium were bought from Clonetics.USA.Mouse anti-CDlla monoclonal andbody,mouse anti-CDllb monoclonal antibody.mouse anti-CD29 monoclenal antibody and mouse anti-CD7l monoclonal antibody Were purchased from Antibody diagnostica,USA.Mouse anti-CD34 monoclonal antibody was obtained from Lab Vision Corporation.USA.METHODS:Umbilical cord blood stem cells were divided into four groups.Umbilical cord blood steTn cells in the group A were incubated in the endothellal basal medium.Umbilical cord blood steHl cells in the group B were incubated in the endothelial basal medium containing microvascular endothelial cells in a 6-well plate.Umbilical cord blood stem cells in the group C were incubated in the endothelial basal medium containing endotheliaI medium in a 6-well plate coated with fibronectin.Umbilical cord blood stelrn cells in the group D were incubated in the endothelial basal medium containing endothelial medium in a 25 cm2 ctdturing flask.MAIN OUTCOME MEASURES:Cell morphology and population doublings were observed.CDlla,CDllb,CD34,CD29 and CD71 expression was detected by immunohistochemistry. RESULTS:Fibroblast-like cells were measured in each group.The celIs grew badly in morphology and proliferated slowly in the group A,while cells proliferated rapidlyinthe group B.The cell clones were instable and inclined to aging in the group C,with a new cell clone was found.The cells in the group D were similar to those in the group C.Surface antigens detected by immunohistochemistry showed CDlla(-),CDllb(-),CD34(-),CD29(+)and CD7l(+). CONCLUSION:MSCs witll well growth and rapid proliferation can be detected in the endothelial basal medium containing endothellal medium in an intact 6-well plate.The outcome is bad in fibronectm-coated plate or 25 cm2 culture flask.
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BACKGROUND: Cell apoptosis and ventricle reconstitution following myocardial infarction are of mutual cause-effect, and they cause vicious cycle. How to reduce the apoptosis events following myocardial infarction is one of keys to saving heart function.OBJECTIVE: To observe the effect of umbilical cord blood mesenchymal stem cell (UCBSMC) transplantation on remaining myocardial tissue of dogs with acute myocardial infarction.DESIGN: A randomized controlled observation.SETTING: Department of Cardiothoracic Surgery, Xinhua Hospital.MATERIALS: This study was carried out in the Central Laboratory of Xinhua Hospital from October 2005 to May 2007.Thirty-six adult hybrid dogs, male and female in half, were provided by the Animal Experimental Center of Xinhua Hospital.METHODS: Thirty-six dogs were divided into cell transplantation group and control group, with 18 dogs in each according to table of random digit. Mesenchymal stem cells were isolated from the umbilical cord blood of full-term pregnant hybrid dogs, cultured and amplified. Then, they were labeled with Laz gene, in vitro induced with 5-azacytidine, and transplanted into the dogs with acute myocardial infarction in the cell transplantation group. Rats in the control group were injected with the same amount of normal saline. Each dog was euthanized by anesthesia for harvesting myocardial specimen 1,4 and 8 weeks after transplantation.MAIN OUTCOME MEASURES: ① Remaining and apoptosis index detected by TUNEL method. ② Myocardial cell volume and histomorphology detected by confocal microscopy. ③ Histological change of myocardial collagen network detected by haematoxylin-basic fuchsin-picric acid staining.RESULTS: Thirty-six involved experimental dogs all entered the stage of final analysis. ①The apoptosis index in the cell transplantation group was significantly lower than that in the control group 1, 4 and 8 weeks after cell transplantation (P ＜0.05). ② Myocardial cell volume in the cell transplantation group 1, 4 and 8 weeks after cell transplantation was significantly larger than that in the control group (P ＜ 0.05). ③ Collagen fiber in the myocardial tissue of dogs in the cell transplantation group was arranged in order and regularly, and in contrast that in the control group was not, and fibers in the control group fused partially.CONCLUSION: UCBSMC transplantation reduces the apoptosis of myocardial cells, promotes the hypertrophy of remaining myocardial cells, regulates myocardial collagen network and improves heart function.
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Objective To study a minimally invasive surgical procedure for coronary artery bypass grafting (GABG). Methods 23 patients, aged from 45 to 73 years old, undervent off-pump coronary artery bypass grafting (OPCABG) according to Octopus procedure from December 1999 to Auguest 2001. In the present study , more than 3 vessels were involved in 19 patients; 2 vessels involved in 3 cases and 1 vessel involved in one case. The grafted arteries included left anterior descending branch (LAD)(23); circumflex(CX)(15); right coronary artery(RCA)(14); ramus intermedius(RUM)(8); and posterior descending artery(PDA) (8). The types of grafts were left internal mammary artery (LIMA)(20) and saphenous vein grafts(SVG)(48). Results None of the patients was managed by means of cardiopulmonary bypass. All survived without angina pectoris. MRI showed that all the conduits were patent postoperatively. Conclusions OPCABG by Octopus procedure can be used in multiple vessel coronary bypass including CX and PDA. The patency rate is similar to that of the conventional technique. The mortality rate and occurrence of complications is low compared with those of the conventional technique . In addition, medical cost is lower. So This procedure is both safe and cost-effective, and is of value in clinical practice.
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Objective Study the prevention and early diagnosis of severe complications of thoracoscopy. Methods 400 patients,(18～81)years,ASA Ⅰ-Ⅲ,underwent thoracoscoy,including 320 pulmonary bubble resections and 80 thorax,mediastinum or lung tumor biopsy or resections.All patients were anesthetized with one-lung ventilation using a double-lumen tube and monitored continuously in circulatory and respiratory functions with a multi-runctional monitor. Results Severe intraoperative complications occurred in 8 patients,including 5 reexpansion pulmonary edema,1 complete occlusion of double-lumen tube due to massive pleural effusion,1 massive bleeding and 1 cardiac arrest due to bilateral pneumothorax.All complications were successfully treated and the patients were cured and discharged. Conclusions Thoracoscopy must be well prepared.Applying one lung anesthesia with double-lumen tube,careful observation during operation,early recognition and treatment of complications may contribute to assuring the safety of this procedure.
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The general clinical abilities of attending doctors are closely linked with the overall quality of patient care in a hospital. A set of quality standards for ward rounds by attending doctors were formulated in the author's hospital and an intensive evaluation was conducted by the experts team of the results of inspection and assessment. It is suggested that the hospital should tighten the quality control of ward rounds by attending doctors and pay serious attention to various problems that may occur. In particular, the hospital should set strict assessment standards for post qualification for attending doctors and strive to cultivate their clinical abilities after their post assumption. In addition, it should bring their talent into full play on the basis of vigorous management and assessment and discover and correct their shortcomings in ward rounds in a timely manner.