Aim To develop a convenient and effective method to isolate and culture primary rabbit aortic vascular smooth muscle cells(VSMCs).Methods The thoracic aortas were removed by dissection under sterile conditions.Aortic smooth muscle cells were excised and cleaned of fat and connective tissue,and the isolated vessel media was cut into 1 mm3 pieces.The explants were digested with different concentrations of collagenase typeⅠ,and incubated at 37℃ for different time,then undispersed explants were placed onto a sterile 100-mm plastic tissue culture dish with growth medium.Results VSMCs could emigrate from the explants digested 6 h by collagenase typeⅠ(1.5 g?L-1)for 24 hours,the cells would passage for another 4~5 days.Confluency could be reached within 3~4 days after subculturing.VSMCs were identified by immunoreactivity with ?-actin and by the smooth muscle cell-specific,hills and valley-like morphology.Conclusion It was an effective method to culture primary VSMCs from the explants digested for 6h by collagenase type Ⅰ(1.5 g?L-1),which could shorten primary culture time.

Aim To develop a convenient and effective method to isolate and culture primary rat aortic endothelial cells (RAECs). Methods The thoracic aortas were removed by dissection under sterile conditions. Aortas were turned over to expose the luminal surface, and the surfaces were digested with different concentrations of collagenase typeⅠ, incubated at 37℃ for different times, then, cut into pieces and placed luminal side down onto collagen-coated flask with growth medium. Results RAECs could emigrate from explants digested 1h by collagenase typeⅠ(2.0 g?L-1) for 24 h and cells would passage for another 4~5 days. Reached confluency within 3~4 d after subculturing. RAECs were identified by immunoreactivity with Factor-Ⅷ and by the endothelial cell-specific, cobblestone-like morphology. Conclusion It is an effective method to culture primary RAECs from explants digested for 1h by collagenase type Ⅰ(2.0 g?L-1),that can shorten primary culture time.

Objective To quantitatively study the correlation of the distance of pulmonary valve to pulmonary artery bifurcation (DPVB) of normal fetuses in second and late trimester with the fetal biometric parameters using fetal echocardiography.Methods The distances of pulmonary valve to pulmonary artery bifurcation in end-systole (DPVBs) and in end-diastole (DPVBd) were measured in a standard short axis view or right ventricular outflow tract view using fetal echocardiography on 419 normal fetuses.The gestation ages ranged from 20 to 35+ weeks.Fetal non-cardiac biometric parameters including biparietal diameter (BPD),femoral length (FL) and gestation age (GA) based on menstrual age,pulmonary valve annulus diameters (PA) at end-systole were measured.The correlation between DPVBs,DPVBd and non-cardiac biometric parameters were analyzed.Results In normal fetuses,the DPVBs was (1.14±0.24)cm with a range of 0.67 to 1.61 cm,the DPVBd was (0.93±0.20)cm with a range of 0.53 to 1.33 cm.The DPVBs and DPVBd were increased with the growth of GA,and were positively correlated with GA,BPD,FL and PA (r=0.827,0.798,0.793,0.769;0.802,0.764,0.773 and 0.771,respectively,all P<0.001).The linear regression equations between DPVBs,DPVBd and GA,BPD,FL,PA were:Y=0.054×GA-0.369,Y=0.184×BPD-0.179,Y=0.217×FL+0.011,Y=1.602×PA+0.156,Y=0.045×GA-0.330,Y=0.152×BPD-0.157,Y=0.182×FL-0.018,Y=1.380×PA+0.080.Conclusions In normal fetuses,the DPVBs and DPVBd increase with the growth of GA,and have good correlation with GA,BPD,FL and PA,respectively.Normal reference ranges of DPVBs and DPVBd have been provided.These normative data may become a new tool for assessment of fetal heart,especially has potential applications in screening of complex congenital heart defects.

Objective To explore the correlation of left ventricular systolic function calculated by automated cardiac motion quantification (aCMQ) and three-dimensional quantitative analysis (3DQA). Methods According to LVEF by 3DQA,patients were divided into abnormal cardiac function group(LVEF<50%)and normal cardiac function group(LVEF≥50%).Dynamic images from two chamber view(AP2), four chamber view(AP4)and three chamber view(AP3)of left ventricular long axis were acquired from 32 patients with abnormal cardiac function and 119 normal subjects.AP2 longitudinal strain (AP2LS),AP4 longitudinal strain (AP4LS) and AP3 longitudinal strain (AP3LS) as well as the left ventricular global longitudinal strain (LVGLS) were measured by aCMQ. While left ventricular end diastolic volume (LVEDV),left ventricular end systolic volume (LVESV) and LVEF were derived through aCMQ automatically calculated region of interest (ROI) driven automation.The correlation of left ventricular systolic function indexes obtained by aCMQ and 3DQA were compared.Results No significant differences were found between the two groups in gender,body mass index(BMI) and age (P > 0.05).LVEF in abnormal group by 3QDA and aCMQ were much lower than those in normal group (P < 0.01).The absolute values of AP2LS,AP4LS,AP3LS and LVGLS in abnormal group were significantly lower than those in normal group (P < 0.01).LVEF by aCMQ in abnormal group was positively correlated with LVEF by 3QDA methods(r =0.91 ,P <0.01);LVEF by aCMQ in normal group was positively correlated with LVEF by 3QDA (r = 0.73,P < 0.01).The left ventricular global longitudinal strain (LVGLS) measured by aCMQ was negatively correlated with LVEF by 3QDA(r = -0.815,P < 0.01).LVEF measured by aCMQ and 3DQA showed high inter-observer and intra-observer agreements in Bland-Altman charts.Conclusions aCMQ has preferable repeatability.Comparing with the traditional measurement method,LVEF measured by aCMQ has higher correlation with that measured by 3QDA.aCMQ can be a new and relatively accurate method to evaluate the left ventricular systolic function.

Objective To determine whether real-time three-dimensional transesophageal echocardiography(RT-3D TEE) is an accurate non-invasive technique for defining the morphology of atrial septal defects(ASD). Methods In 20 patients with secundum ASD, mean age 42.2 years (7 male, 13 female) ,live three dimensional surgical views of ASD were acquired,qualitative morphology of ASD such as the shape, presence of fenestrations and the defect margins were noted during the cardiac cycle. The measurements obtained from 2-dimensional transesophageal echocardiography(2D-TEE) and RT-3D TEE were compared to those obtained from stretched balloon diameter(SBD) or surgery. Measurements of the size and area change of ASD were validated during the cardiac cycle by RT-3D TEE,stepwise multiple linear regression analysis was performed to test the correlation between the maximum change ratios of area and diameter of ASD and age, atrial septal length, ASD shunting velocity, the maximum diameter of ASD and right ventricular systolic pressure. Results ① The morphology of ASD was circle-like or elliptical in systole,irregular or elliptical in diastole. ②The area and diameter of ASD measured by RT-3D TEE was minimal in isovolumetric contraction phase, maximal in isovolumetric diastole phase. The correlation coefficient of maximum diameter measured by 3D with SBD was 0.962 , greater than that by 2D and SBD (0.820). ③The change ratio of area and maximum diameter of ASD acquired by RT-3D-TEE was 11.48%-71.12% and 2.80%-43.87% respectively,and the correlation coefficient of them was 0.921. Conclusions RT-3D TEE using live 3D-Zoom mode accurately displayed the varying morphology,dimensions and spatial relations of ASD. RT-3D TEE can offer visualization morphological changing of ASD in different periods of cardiac cycle,providing more information for percutanous catheter intervention and open heart surgery.

Objective To evaluate left ventricular rotation and torsion and its early recovery after percutaneous coronary intervention (PCI) in patients with coronary heart disease and normal left ventricular ejection fraction (LVEF) using two dimensional speckle tracking echocardiography.Methods Twenty three consecutive patients with coronary heart disease and normal LVEF were divided into group B (with coronary stenosis ＜70％) and group C (with coronary stenosis ＞70％ and with PCI).Along with 11 healthy controls(group A),indices including basal rotation (BR),apical rotation (AR),left ventricular torsion (LVT) and normalized time to peak were compared among groups,correlative analysis was made between LVT and each indices mentioned above,indices of group C before and 24 hours after PCI were compared.Results AR,LVT in group B and C reduced relative to group A (P ＜0.05),meanwhile time to peak of BR in group C shortened relative to other groups.BR,AR and normalized time to peak of BR were correlated to LVT respectively.BR and LVT in group C increased after PCI(P ＜0.05).Conclusions AR was sensitive to ischemia,the reduction of time to peak of BR in group C might be restriction and compensation.Sensitive to early recovery of left ventricular function after reperfusion,BR could be a predictive index of early effect of percutaneous coronary intervention.

Objective To investigate the value of the intraoperative real-time three-dimensional transesophageal echocardiography (RT 3D-TEE) in therapeutic application of surgery for the mitral valve prolapse.Methods Thirty-five patients underwent surgical treatment were diagnosed as mitral valve prolapse by 2-dimensional transthoracic echocardiography (2D-TTE),with or without ruptured chordae tendineae.RT 3D-TEE was performed for assessing the segment of prolapse and its complications before cardiopulmonary bypass and after heart resuscitation respectively.The echocardiographic results were compared with the surgical findings.Results The accuracy of RT 3D-TEE to identify segments with prolapse was significantly higher than that of 2D-TTE (91.4％ vs 82.9％,P =0.003).RT 3D-TEE was more accurate than 2D-TTE for identifying ruptured chordae tendineae,vegetation and thrombus since higher sensitivity,specificity and Youden index.Among all 35 patients,valvuloplasty was performed in 28 cases,whereas,only 7 cases underwent replacement of prosthetic valves.Three patients were benefited by accurate evaluation of therapeutic effect using RT 3D-TEE in time.ConclusionsIntraoperative RT 3D-TEE can be used to provide “ surgical view”,acquired adequate valuable information of mitral valve,and more reliable functional and anatomical assessment of the mitral valve components and its geometry.Suitable surgical intervention can be designed for improving outcomes of patients.

ObjectiveTo quantitative evaluate the alterations of cardiac morphology and function in gestational impaired glucose tolerance(GIGT) fetuses.MethodsFetal echocardiograms were performed on 68 GIGT fetuses with gestation age between 21 ～ 40 weeks for evaluation of cardiac morphology and function.Fetal cardiac morphology,systolic and diastolic functions of 68 GIGT fetuses were compared with 81 control group fetuses using conventional two-dimensional,M-mode,pulsed Doppler echocardiography and myocardial tissue Doppler imaging.ResultsComprehensive fetal echocardiography data analysis showed no significant differences in cardiac morphology and function between two groups (P＞0.05).Conclusions The alterations of cardiac morphology and function in GIGT fetuses can be accurately and objectively evaluated using quantitative evaluation in fetal echocardiography and will help to offer consultation.

Objective To establish reference values of normal fetal heart conduction time intervals by tissue Doppler imaging(TDI)and pulsed Doppler(PD)echocardiography,and to assess their correlation with gestational-age and fetal heart rate.Methods One hundred and eighty-nine pregnant women underwent detailed echocardiographic examinations.Atrio-ventricular conduction time interval(AV)and the time interval from onset of ventricular contraction to the onset of atrial contraction of next cardiac cycle(VA)were measured by TDI and PD echocardiography.Results TDI-AV was(126.56±15.33)ms(95% CI 124.10～129.03 ms),TDI-VA was(285.22±24.53)ms(95% CI 281.27～289.16 ms),PD-AV was(127.42±12.88)ms(95% CI 125.35～129.49 ms),PD-VA was(287.42±25.19)ms(95% CI 283.37～291.47 ms).A paired t test revealed no systematic difference between the two approaches used to measure AV and VA.AV and VA were significantly positively correlated with gestational age,and significantly negatively correlated with fetal heart rate.Heart conduction time intervals were altered in fetus with paroxysmal arrhythmia.Conclusions This study established the normal values of fetal heart conduction time intervals.Prenatal determination of fetal heart conductional time intervals has important potential clinical utility in assessing fetal arrhythmia.

Objective To determine the usefulness of real time three-dimensional transesophageal echocardiography(RT-3D TEE) in the preoperative assessment of mitral valve (MV) pathology by comparing images with surgical findings and to evaluate the function of MV postoperatively. Methods Nineteen consecutive adult patients with established diagnosis of mitral regurgitation(MR) scheduled for surgical correction were enrolled. Intraoperative 2D and 3D transesophageal echocardiography(TEE) were performed. All the 3D images were compared with findings obtained from direct surgical inspection. Postoperative RT-3D TEE was performed immediately to evaluate outcomes of mitral annuloplasty. Results Superb 3D-TEE en face views of the MV were obtained in all patients. Correct diagnoses of MV pathology in agreement with the surgical findings were made in 16 of 19 patients (84. 2%), however in the remaining 3 patients, the diagnoses were incorrect despite good image quality. In one, RT-3D TEE diagnosed prolapse of A2 segment and P2 scallop,which was not confirmed at surgery, but MV degeneration and annular dilatation were observed. In another one, RT-3D TEE revealed prolapse of P3 scallop, while at surgery prolapse involved P2 and P3 with ruptured chordae. In the rest one,surgically detected prolapse of P2 was missed by RT-3D TEE. On the other hand with 2D TEE,the diagnoses correlated poorly with surgical findings,only 10 patients were accurately diagnosed (52.6%). In some patients,2D TEE was able to identify the MVP, but it had difficulty in defining the exact location of the prolapsed segment or scallop. The severity of MR decreased significantly after surgery. MV repair was successful in all patients except one(5.1%), in whom moderate to severe MR was still present and MV replacement was conducted subsequently as an alternative.Conclusions RT-3D TEE provides excellent imaging of MV components, including the anterior and posterior leaflets, as well as annulus and subvalvular structures, which can be especially helpful in planning the most appropriate surgery strategy. RT-3D TEE offers exact anatomic characteristics of mitral annuloplasty rings and bands, providing additional information for the evaluation of surgical outcomes.