BACKGROUND: Under certain condition, circulating endothelial progenitor cells (EPCs) can differentiate into endothelial cells, and further participate in angiogenesis.OBJECTIVE: The goal of this study was to investigate the feasibility and efficacy of peripheral blood-derived EPCs in promoting angiogenesis in the ischemic myocardium, in order to provide a new cell implanting method for the treatment of coronary heart disease.DESIGN: A randomized controlled experiment.MATERIALS: Sixty male Sprague-Dawley (SD) rats, of clean grade, weighing (340±20) g, were provided by Qingdao Laboratory Animal Center. These animals were randomly divided into 2 groups with 30 rats in each: experimental group and control group. In each group, ten rats were separately observed 2, 4 and 8 weeks after EPCs being injected. The protocol was conducted in accordance with animal ethics guidelines for the use and care of animals.METHODS: This study was carried out in the Qingdao Key Laboratory of Medical Biological Technology between May 2003 and September 2004. After SD rats in the experimental group were anesthetized, peripheral blood was taken.Mononuclear cells were harvested by density gradient centrifugation. CD31, CD34, Flk-1 and von Willebrand disease factor immunofluorescence staining positive EPCs were harvested by adding the defined media of vascular endothelial growth factors and basic fibroblast growth factor. Myocardial ischemia was induced by ligation of murine left anterior descending coronary artery. Autologous EPCs isolated from the peripheral blood of each animal were infused to ischemic myocardium. In the control groups,cell culture media were infused, and the other procedures were the same as those in the experimental group. Two, four and eight weeks after ligation, all animals were sacrificed by overdose anesthesia, and heart tissue sections were made.MAIN OUTCOME MEASURES: ①After haematoxylin-eosin staining, myocardial structure changes were observed under an optical microscope. ②Cells positive for factor Ⅷ were numbered and then to calculate the total number of cells in each visual field for evaluating microvessel density with a German ZEISS Axiotron image analyzer.RESULTS: All the 60 SD rats were involved in the final analysis, without deletion. ①Myocardial structure was relatively disarrayed in the control groups, with collagens and fibroblasts substituted for cardiomyocytes. In the marginal infarct areas, cardiomyocytes were irregular and partial cells were significantly hypertrophied. As compared with control group, myocardial collagen fibers fused less, and tissue structure got more disarranged in the experimental group. Microvessel density in the implanted area was obviously increased. ②In the 2nd, 4th and 8th weeks after EPC infusion, microvessel density at the ischemic myocardium was significantly higher than that at the corresponding time point in the control group (P ＜ 0.01). In the EPCs groups, microvessel density tended to statistically increase with time going (P ＜ 0.05). No prominent difference was observed in the three control groups (P＞ 0.05).CONCLUSION: Relatively purified EPCs can be obtained by certain procedure of isolation and culture from rat peripheral blood. Intramyocardial implantation of autologous EPCs promotes microangiogenesis and has a protective effect on ischemic myocardial tissue.

Objective To compare the treatment outcomes between minimally invasive perventricular device occlusion (MIPDO) and right subaxillary incision surgical repair(RSISR) on perimembranous ventricular septal defect(PmVSD) in children less than 15 kilograms.Methods From January,2010 to January,2013,a total of 530 infants(age ＜ 3 years,weigh ＜ 15 kg) with PmVSD enrolled and they were divided into two groups according to different treatment methods at random.Group 1 (265 cases) was arranged perventricular device closure with modified occluders through a lower partial median sternotomy under transesophageal echocardiography (TEE) guidance;group 2 (265 cases) was arranged surgical repair on cardiopulmonary bypass(CPB) through a right subaxillary small incision.A prospective randomized controlled study was performed between two groups on success rate,operation time,volume of blood loss and transfusion,length of intubation and ICU stay,complications,expenses and follow-up results etc.Results All patients in two groups obtained effective treatment with no death or serious life-threatening complications.Group 1:255 cases (96.23％) underwent successfully MIPDO.The remainder 10 cases (3.77％) who failed in attempt were successfully converted to conventional open heart operation by extending the original incision.Different arrhythmias arose in 30 cases(11.76％),including incomplete left bundle branch block(ILBB) in 3 cases(1.18％),complete right bundle branch block(CRBB) in 3 cases(1.18％),incomplete right bundle branch block(IRBB) in 16 cases(6.27％),Ⅰ° atrioventricular block(Ⅰ°AVB) in 8 cases(3.14％);trivial residual shunt(RS) in 18 cases(7.06％);newly arose trivial tricuspid regurgitation(TR) in 29 cases(11.37％).Group 2:All the patients(100％) underwent successful surgical repair through right subaxillary incision.Different arrhythmias occurred in 116 cases (43.77％),including transient complete atrioventricular block(CAVB) and ILBB in 2 cases respective(0.75％),junctional ectopic tachycardia(JET) in 1 cases(0.38％),CRBB in 61 cases(23.02％),IRBB in 52 cases(19.62％);trivial RS in 16 cases (6.04％);newly arose trivial TR in 11 cases(4.15％);heart dysfunction in 17 patients(6.42％).All patients were followed up for more than 12 months,and there were no newly happened or aggravated valve regurgitation or late onset CAVB in two groups.The final treatment effects are similar in both groups.But group 1 was significantly superior to group 2 in the aspects of operation time,volume of blood loss and consumption,length of intubation and ICU stay,hospitalizations and costs(all P ＜ 0.05).The incidence of TR is higher in group 1 (P ＜ 0.05),and that of right bundle branch block was higher in group 2 (P ＜ 0.05).The incision is longer in group 2,but in a less exposed location.CPB is not needed in group 1,but anticoagulant drug is required for 3-6 months.Conclusion Both RSISR and MIPDO are effective treatment methods of PmVSD.Though having some limitations,MIPDO which characterized by simple procedure,minimal invasion,quick recovery,saving of medical resources could not only minimize the surgical trauma to patients,but also ensure the safety of operation to the maximum extent.However,the patient selection is vital.For selected patients,especially those of moderate PmVSDs with obvious clinical symptoms but no cardiac valve regurgitation,it is an ideal approach.

Objective To explore the procedures of isolation,cultivation of endothelial progenitor cells(EPC) from peripheral bloods and the feasibility of local EPC transplantation to promote neovascularization in ischemic myocardium. Methods Peripheral blood was obtained from the femoral artery of each Sprague-Dawley(SD)rat.Myocardial ischemia was induced by ligation of murine left anterior descending coronary artery.Immmdiate timepoint of ligation procedure, autologous EPC from peripheral blood,positive for CD31,D34,Flk-1 and vWF,isolated from the peripheral blood of each animal were injected to ischemic myocardium.In the control groups,cell culture media was injected.Rats were euthanized and neovascularization in local ischemia areas was evaluated.Results Hematoxylin-eosin staining indicated better myocardial arrangement in the EPC group.Capillary density in ischemic myocardium weeks after transplantation was significantly greater in the group of EPC injection than the control groups.Conclusion Relatively purified EPC can be obtained by certain procedure of isolation and culture from rat peripheral blood.Intramyocardial transplantation of autologous EPCs will promote microangiogenesis and has a protective effect on ischemic myocardial tissue.

Objective To review our clinical experience with primary surgical repair of tetralogy of Fallot in neonates and premature infants and to discuss the timing of repair and major factors in treating this patients perioperatively.Methods From January 2012 to September 2015,a series of 19 consecutive neonates and premature infants(M/F =12/7) with tetralogy of Fallot were admitted in our center to receive surgical treatment,with a mean age of(17.3 ± 5.5) days(12-28 days) and a mean body weight of(2.9 ±0.7) kg(2.1-4.3 kg).All the 19 cases were symptomatic with cyanosis,saturation on room air 0.79 ± 0.12 (0.48-0.92),and shortness of breath.Before operation,2 cases were receiving an infusion of prostaglandin E1,5 were mechanically ventilated.7 were more than moderate anemia with hemoglobin of 55-87 g/L.All the patients received echocardiography,ECG and chest X-ray.The McGoon ratio and Nakata index were 1.09 ± 0.30 (0.8 to 1.6) and (135.5 ± 54.2) mm2/m2 (63-212 mm2/m2) respectively.18 cases received one-stage surgical repair and 1 premature infant under two-stage operation with the VSD closure after right ventricular outflow tract(RVOT) transannular patch augmentation.All the VSDs in the 18 cases were closed with continuous suture and RVOT were enlarged with autologous pericardium patch transannularly or not.Balanced and modified ultrafiltration were applied in all the patients.Patients were strictly followed up with a standard protocol focusing on right ventricular function and arrythmia.Several characteristics(e.g.time of operating,mechanical ventilation and ICU stay,complications,hospital stay time,cost of hospitalization) were compared between this group of patients and other TOF patients during the same period in our center.Results All the one-stage operations were successful.There was no mortality and major complication.Mean CPB and aortic clamping time were(111.5 ± 31.6)min (76-153min) and (73.3 ± 11.6) min (64-89 min) respectively.10 VSDs were closed with transventricular approach,6 witht transatrial appraoch and 2 with transatrial-ventricular approach.12 cases (66.7 ％) had a transannular RVOT patch,4 (22.2 ％) with single RVOT patch and 2 (11.1％) with transannular RVOT and left pulmonary artery patch.Atrial communication were left open in 15 cases (83.3 ％).The time of mechincal ventilation and ICU stay were (123.7 ± 59.5) h (39-239 h) and (10.1 ± 3.2) days (5-19 days) respectively.All the patients were followed up for (31.8 ± 15.7) months (9-57 months).There was no mortality and major complication.ECG showed that there was no severe arrythmia except for 3 complete right branh bundle block.The latest echocardiography results showed that right heart function was normal in all the cases and RVOT grandients was less than 30mmHg except one with 35 mmHg.There were 2 cases with residual shunt less than 2 mm and 8 cases with pulmonary valvular regurgitation less than moderate degree.Compared with other TOF cases during the same period,there was no difference according to the data mentioned above except with more time of mechnical ventilation and ICU stay and more cost of hospitalization.Conclusion Primary repair of TOF can be performed safely in symptomatic neonates and premature infants,regardless of age and body weight,with favorable early and mid-term results.Excellent teamwork and accurate prenatal and postnatal diagnosis were the two major factors in yeilding good results in these patients.

Objective To discuss one-stage surgical repair of interrupted aortic arch ( IAA) in neonates and young in-fants, summarize follow-up results and subsequent treatments.Methods From September 2010 to December 2014, a series of 38 consecutive neonates and young infants ( M/F =26/12 ) with IAA were admitted in our surgical group to receive surgical treatment, with a median age of 11 days(1 day to 5 months) and a median body weight of 4.1 kg(1.8 to 5.8 kg).IAA was type A in 24 cases, type B in 13 cases and type C in 1 case.Associated cadiovascular anomalies were common except 2 cases, including atrial septal defect(n=23), ventricular septal defect(n=36), left ventricular outlet tract obstruction(LVOTO)(n=7), anomalous left pulmonary artery from aorta(n=1), truncus arteriosus(n=1).There were 5 cases of airway stenosis in 28 cases of CT angiography and 3D airway reconstruction.All the 38 cases were admitted with differential cyanosis.36 cases were recieved one-stage surgical repair except 2 cases died before operation.35 cases of IAA associated with intracardiac anom-alies were repaired through a median sernotomy.Cardiopulmonary bypass(CPB) was performed with two canulations in ascend-ing aorta and main pulmonary artery.Selective cerebral perfusion(n=21) or circulatory arrest(n =14) was carried out with body temperature of 18℃-23℃.After the patent ductus arteriosus tissue was completely resected , a continuous end-to-end or end-to-side suturing with 7/0(8/0) prolene was performed.In 1 type B cases with expected excessive anastomotic tension, the left subclavian artery was cut off to release the aortic arch .The associated anomalies were repaired during the cooling or re-warming period.Chest close was delayed to carry out no more than 24-72 h postoperatively in 16 cases.The other one type A IAA case without intracardiac anomaly was repaired through a standard left posterior lateral incision .Patients were strictly fol-lowed up with a standard protocol.Complications such as aortic arch restenosis, new LVOTO, new airway stenosis, and so on, were specialy focused on during the follow-up period.If needed, reoperation or transcatheter intervention was applied to treat the complications.Results Two cases died before operation and another 2 died in the early period postoperatively because of pulmonary hypertension crisis and severe tracheal stenosis, respectively.Sudden death was in 1 case during the follow-up peri-od.Median CPB time was 138 min(105-208 min) and median selective cerebral perfusion or circulatory arrest time was 24 min(16-35 min) .Mechanical ventilation time was 2-25 days.Blood pressure difference between the upper and lower limbs was less than 20 mmHg in all the patients before discharge.29 cases(85.29%) was followed up for 28 months(9 -60 months).3 cases were received reoperation in 5 cases of newly detected LVOTO.One case received reoperation in the 7 preop-erative LVOTO cases because of newly detected LVOTO(blood pressure gradient more than 40 mmHg).Two cases with aortic arch restenosis received percutaneous transcatheter ballon dilation .One case with new left main bronchus stenosis after opera-tion was strictly followed up.Conclusion IAA is a rare and severe congenital heart disease.Positive surgical repair should be performed after definite diagnosis being made .Although satisfactory results can be obtained with one-stage primary operation , IAA is a kind of progressive chronic disease.New postoperative LVOTO should be focused on becasue it will need reoperation or even the third operation .

Objective: To summarize the experience for treating pulmonary venous obstruction in patients after total anomalous pulmonary venous connection (TAPVC) operation. Methods: A total of 16 patients with post-TAPVC pulmonary venous obstruction in our hospital from 2011-01 to 2015-12 were retrospectively analyzed including10 male. All patients received echocardiography, electrocardiogram and chest X-ray examinations at pre-discharge, 1, 3, 6, 12 and 24 months post-operation. Pulmonary venous obstruction was diagnosed by echocardiography measured pulmonary vein (PV) lfow speed>2m/s. The time of re-operation was determined by clinical manifestations as recurrent heart failure and growth retardation; sutureless technique and conventional patch enlarge technique were used in the second operation. Results: No one lost contact in all 16 patients. There were 7/16 patients with anastomotic stenosis (1 mixed type, 3 infracardiac type, 2 supracardiac type and 1 cardiac type), 7 patients with one PV stenosis, 2 with two PV stenosis and nobody with three or more PV stenosis. Based on per-operative Darling classiifcation, there were 2 patients with mixed type, 5 with infracardiac type, 5 with supracardiac type and 4 with cardiac type. Most post-operative PV stenosis occurred at 3-6 months after the surgery. There were 5 patients receive re-operation, 4 with sutureless technique, 1 with conventional patch enlarge technique and all of them suffered from anastomotic stenosis. 2 patients died and 3 were followed-up. Conclusion: Post-operative anastomotic stenosis was the main indication for re-operation in patients after TAPVC; early operation could better improve the clinical condition.

Objective To observe the availability and safety of ventricular septal defect (VSD) occluder in infants and young children with large patent ductus arteriosus (PDA) associated with severe pulmonary hypertension.Methods Five patients (1 male and 4 fomale) of large PDA aged 5 months to 3 years,weighted from 5.1 to 15 kg,body surface area (BSA) 0.37-0.58 m2 underwent transcathter intervention with concentric VSD occluders from June 2008 to May 2009.Arterial ducta were tube-like and their diameters were 5.7 to 8.5 mm,with ulmonary vascular resistance from 4.8 to 5.7 Wood Unit,Qp/Qs 3.4-4.6.Three patients were given Bosentan after intervention.Results The large PDAs were successfully closed with VSD occluders,including 1 concentric perimembranous VSD occluder and 4 muscular VSD occluders.They all discharged 4 to 5 days with hidrosis and weight improved.Echocardiogram indicated VSD occluder was stable,no residue shunt and no stricture of left pulmonary artery and descending aorta were found.According to tricuspid and pulmonary regurgitation,pulmonary arterial pressure decreased differently and returned to normal after 6 months follow-up.Conclusion VSD occluder is available and effective to close large PDA associated with severe pulmonary hypertension in inrants and young children,but more cases and long-term follow-up are necessary.

Objective Transesophageal echocardiography (TEE) guided, minimally invasive perventricular device occlusion of ventricular septal defects ( VSDs) without cardiopulmonary bypass ( CPB) has been applied in multiple centers. We reported experiences and the mid-term results. Methods Four hundred and thirty-two cases from 4 cardiac centers were involved in the study. There were 235 males and 197 females, aged from 3 months to 15 years, with a body weight varying from 4.0 to 26.0 kg. Three hundred and fifty-one patients had perimembranous VSDs, 57 had intracristal or supracristal VSDs and 24 had muscular VSDs (17 had multiple muscular VSDs). The diameter of the VSD ranged from 3 to 12 (5.3 ±1.6 ) mm.For those with perimembranous or muscular VSDs, a 3 to 5 cm inferior sternotomy was made, but for those with intracristal or supracristal VSDs, a 2 to 3 cm incision was made parastemally through the left third intercostal space. Being monitored and guided with TEE, the device was deployed to occlude the VSD through the puncture at the free wall of the right ventricle. TEE was used for assessing the residual shunting, the left and right ventricular outlet tracts, valvular function and for detecting any arrhythmia, The devices would be released if the heart rhythm was normal, as well as the residual shunting and valvular regurgilalion were not detected. Results The procedure was completed successfully in 417 cases(96.5% ) and converted to traditional surgical closure with CPB in the other 15 cases(3.5% ). Concentric devices were used in 238 cases(57.1% )and eccentric devices were used in 179 patients(42.9% ). Successful procedures finished in less than 90 minutes, and the deployment and evaluation of the devices were completed in 5 to 60 (18. 2 ± 8.6) minutes. No residual shunt and detectable aortic or tricuspid insufficiency and arrhythmia was observed. Patients were extubated within 2 hours and discharged 3 to 5 days after the operation. During fellow-up period from 3 months to 2 years, no clinically significant complications occurred. Conclusion The minimally invasive device closure of VSD under TEE guidance without CPB is proved to be a simple, safe and effective treatment for a considerable number of children with VSD. Its use in the clinical practice should be encouraged.

OBJECTIVETo evaluate the effect of milkvetch injection (MI) on immune function of children with tetralogy of Fallot (TOF) after radical operation.

METHODSForty-children with TOF were divided into two groups, the 20 in the control group treated with conventional treatment alone and the 20 in the treated group treated with conventional treatment plus 15 ml of MI every 12 hrs for 14 days. Changes of immunoglobulin, complements, lymphocyte phenotypes and cytokines were observed.

RESULTSIn the treated group, the abnormally increased levels of IgG, IgM, C3, C4, CD8+ and CD19+ began to lower at lst-2nd week after treatment, and basically restored to the levels of normal at 3rd-4th week; while the decreased levels of IgA, CD3+, CD4+, CD4+/CD8+ ratio, CD3+/HLA-DR+ and CD3+/CD16+ -CD56+ raised gradually from the 1st week and restored to normal range at 2nd-3rd week. The IL-6 and tumor necrosis factor-alpha (TNF-alpha) levels in the plasma and supernatant, produced in vitro by peripheral blood mononuclear cells (PBMC) decreased gradually at 1st week and restored to the normal level at 3rd-4th weeks. The different value before and after treatment of the above-mentioned indexes in the treated group were superior to those in the control group (P<0.05 or P<0.01).

CONCLUSIONMI could significantly improve the immune function of children with TOF after radical operation.

Adjuvants, Immunologic ; therapeutic use ; Astragalus Plant ; CD4-CD8 Ratio ; Cardiopulmonary Bypass ; Child ; Child, Preschool ; Complement C4 ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulin G ; blood ; Infusions, Intravenous ; Male ; Phytotherapy ; Postoperative Period ; Tetralogy of Fallot ; drug therapy ; immunology ; surgery ; Tumor Necrosis Factor-alpha ; metabolism