Mesenchymal stem cells and endothelial progenitor cells obtained from rabbit bone marrow with differential adhesion methods and their biological characteristics / 中国实验血液学杂志

This study was aimed to evaluate an effective and stable method for obtaining mesenchymal stem cells (MSC) and endothelial progenitor cells (EPC) from the rabbit bone marrow and to investigate the biological characteristics of MSC and EPC. Mononuclear cells were obtained from rabbit bone marrow using density gradient method, and were differentially adhered to the cell culture plate enclosed with fibronectin. Then, MSC and EPC were amplified with EGB-2MV medium. Trypan blue method was used to test the passage survival rate. Growth curve, MTT and DNA cycle were used to evaluate the proliferation ability of MSC and EPC. MSC were identified with induced differentiation into the osteoblasts and adipocytes, and their immune phenotype was detected by flow cytometry (FCM). EPC were characterized by the special digestion of Dil-ac-LDL, FITC-UEA-I, and the conjunction with CD133, VEGFR2/KDR and CD34, their purity was also calculated. The results indicated that the colony was obviously formed when the mononuclear cells were cultured for 24 hours and, 80% of the cells became long spindle and integrated at d 8. Cells, which were adhered for twice, were cultured with EGM-2MV medium, began to extend at d 3, and became strip-shaped and integrated for about 80% at d 8. Passage survival rates were more than 90% for both cells, and after passage 2 the growth curve was like "S". Optical density was changed obviously when the cells were cultured for 3 - 5 d, but there were no significant difference of cell cycles between MSC and EPC, which G0-G1 was (93.32 ± 1.65)% and (93.05 ± 1.95)% respectively. Positive rates were (99.7 ± 1.12)%, (99.1 ± 2.33)%, (4.8 ± 0.38)%, (6.8 ± 0.49)% and (0.4 ± 0.08)% for CD90, CD44, CD14, CD45 and CD79a respectively. MSC were identified by induced differentiation into osteoblasts and adipocytes. Positive rates of the EPC, which were adhered for twice and passaged 2, were (82.1 ± 3.4)% for fluorescent staining of Dil-ac-LDL and FITC-UEA-I, and (74.2 ± 3.2)%, (64.7 ± 4.3)% and (43.5 ± 1.5)% for CD133, VEGFR2/KDR and CD34 respectively. It is concluded that high-purity MSC can be obtained with density gradient and differential adhesion method, and high-proliferation EPC can be cultured with EGM-2MV medium in cell plates enclosed with fibronectin, so they may become the optimal seed cells for tissue engineering study.

An in vivo model of in situ implantation using pulmonary valved conduit in large animals under off-pump condition / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The application of pulmonary valved conduit to reconstruct the continuity between right ventricles and pulmonary artery is one of the major surgeries. This study aimed to establish an in vivo model of in situ implantation using pulmonary valved conduit in large animals under off-pump condition to validate the long-term effects of artificial pulmonary valved conduit.</p><p><b>METHODS</b>Domesticate juvenile male sheep and tissue-engineered porcine pulmonary valved conduit were used for the experiment: 30 sheep, weighing (15 ± 3) kg (range 13 to 17 kg) were randomly divided into two groups which were all operated under general anesthesia by off-pump surgery (group 1) and left thoracotomy (group 2). Two different off-pump surgical methods were used to perform cannulation in sheep pulmonary artery to replace part of sheep pulmonary artery with pulmonary valved conduit which will work together with sheep pulmonary artery and valves. During the experiments, animal survival, complication rates, operating time and blood loss were recorded to compare the results between groups and to establish a surgical method with minimal invasion, simplicity, safety, and high success rates.</p><p><b>RESULTS</b>In group 1, a total of 15 cases of surgeries were performed, in which two sheep died; the operative mortality was 13.3% (2/15). In group 2, a total of 15 cases of surgeries were performed, and the surgical mortality rate was 0 (0/15). The operation time and blood loss in group 2 was significantly better than that in group 1. The postoperative echocardiograms showed that, after the surgeries by these two methods, the blood flows were normal, and the valves can open and close freely. Autopsy after 6 months showed that the inner wall and the valves of pulmonary valved conduit were smooth with no thrombus formation.</p><p><b>CONCLUSION</b>These two off-pump methods are feasible and safe with fewer traumas; but the second method is better and particularly suitable for the establishment of a juvenile animal model.</p>

Gallbladder Tuberculosis: CT Findings with Histopathologic Correlation

OBJECTIVE: We wanted to describe the computed tomography (CT) findings of gallbladder tuberculosis (TB) and to correlate them with pathologic findings. MATERIALS AND METHODS: There were seven patients (M:F = 3:4; mean age, 46.3 years; age range, 32 to 78 years) in whom gallbladder TB was eventually diagnosed. All of them underwent cross-sectional imaging with CT, a pathologic examination and a retrospective review. CT imaging evaluation was done in each case, including the findings of a mass versus nodule, wall thickening (uniform or irregular) and the enhancement patterns (homogeneous or heterogeneous). RESULTS: All the cases of gallbladder TB revealed the following three different CT findings: micronodular lesion of the gallbladder wall (n = 1), a thickened wall (n = 4) and a gallbladder mass (n = 2). There were three cases of homogeneous enhancement of the lesions, including homogeneous enhancement with nodular lesion, homogeneous uniform thickness enhancement and homogeneous thickness enhancement in one case each, and these cases pathology showed tuberculous granuloma with a little caseating necrosis in one case and tuberculous granuloma with rich fibrous tissue, but little or no evident caseating necrosis in two cases. Four cases of heterogeneous enhancement of the lesions, including heterogeneous uniform-thickness enhancement in two cases, heterogeneous enhancement with a local mass lesion in one case and heterogeneous enhancement with a mass that replaced the gallbladder in one case; in these cases, pathology showed tuberculous granuloma with marked caseation or liquefaction necrosis in three cases and tuberculous granuloma by fibrous and calcifications accompanied by caseating necrosis in one case. Among the seven cases of gallbladder TB, six cases were accompanied by abdominal extra-gallbladder TB, including abdominal lymph node TB in five cases and hepatic TB in four cases. CONCLUSION: Gallbladder TB has various CT manifestations, and the enhanced CT findings are well matched with pathological features. An irregularly thickened gallbladder wall or a gallbladder wall mass with multiple-focus necrosis or calcifications accompanied by the typical CT findings of abdominal extra-gallbladder TB should suggest the diagnosis of gallbladder TB.

Biochemical metabolic changes detected by phosphorus-31 MR spectroscopy in liver of fasting rabbits / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the biochemical metabolic changes detected by phosphorus-31 MR spectroscopy ((31)P MRS) with pathologic changes in the liver of fasting rabbits.</p><p><b>METHODS</b>A total of 22 rabbits were under the starvation up to death to establish animal models. Hepatic (31)P MRS was performed in different period of 10 rabbits including normal condition, over-starvation, agonal condition and death after 30 min. Other 9 rabbits were divided into three type including over-starvation, agonal condition and death group with 3 rabbits in each group, and 3 healthy rabbits served as controls. All the 12 rabbits were sacrificed for the hepatic pathological examination. The MR examination was performed on a 1.5 T imager using a 1H/31P surface coil by the 2D chemical shift imaging technique. The relative quantities of phosphomonoesters (PME), phosphodiesters (PDE), inorganic phosphate (Pi) and adenosine triphosphate (ATP) were measured.</p><p><b>RESULTS</b>All the relative quantification of phosphorus metabolites were changed significantly from starvation to death (X(2)=23.13-35.41, P<0.01). The relative quantifications of ATP of normal condition, over-starvation, agonal condition and death were 2.54 +/-0.53, 1.73 +/-0.14, 0.88 +/-0.23 and 0.05 +/-0.08, respectively (rs=1.0, P<0.01). The relative quantifications of PDE from normal to death were 1.25 +/-0.54, 2.76 +/-0.23, 3.33 +/-0.49 and 3.87 +/-0.43, respectively, and those of Pi were 0.42 +/-0.02, 0.65 +/-0.05, 0.89 +/-0.15 and 0.99 +/-0.08, respectively (rs=1.0, P <0.01). The relative quantifications of PME were also significantly changed (rs=0.4, P=0.6). The pathologic changes of normal condition, over-starvation, agonal condition and death: decreased size of hepatocytes, loss of cell number, cellular swelling, degeneration and cell necrosis or hepatic hemorrhage became more and more pronounced.</p><p><b>CONCLUSION</b>(31)P MRS can monitor dynamic changes of relative quantification of phosphorus metabolites, which are correlated with the pathological severity of acute hepatic injury by fasting.</p>

Evaluation of biochemical metabolites by 31P MR spectroscopy in leukemic infiltration of liver / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the changes of phosphorus metabolites in leukemic infiltration of liver (LIL) with two-dimensional chemical shift imaging(2D CSI)(31)phosphorus MR spectroscopy ((31)P MRS).</p><p><b>METHODS</b>Fifteen patients with LIL and 12 healthy subjects (control group) were scanned with liver 2D CSI(31)P MRS by a 1.5T MR Scanner(Sonata, Siemens Corporation). Relative quantification of phosphorus metabolites including phosphomonoesters (PME), inorganic phosphate (Pi), phosphodiesters (PDE) and beta-adenosine- triphosphate (beta-ATP) were detected and after calibrated with model factor, the ratios of PME/PDE, PME/(PME+PDE), PME/ATP, PDE/ATP and Pi/ATP were analyzed.</p><p><b>RESULTS</b>Compared with control group, the PME value, PME/PDE ratio, PME/(PME+PDE) ratio and PME/ATP ratio were increased in LIL group (1.992 +/-0.876 Compared with 1.167 +/-0.427, P <0.05), (0.551 +/-0.339 Compared with 0.254 +/-0.059,P <0.01), (0.326 +/-0.13 Compared with 0.199 +/-0.049, P <0.01)and (1.402 +/-0.654 Compared with 0.792 +/-0.232, P <0.01) respectively.</p><p><b>CONCLUSION</b>(31)P MRS examination can be used as a non-invasive procedure to evaluate the changes of phosphorus metabolites of leukemic infiltration of liver. The increase of PME value and its ratios to PDE, ATP and PME+PDE may indicate leukemic infiltration of liver.</p>

Effect of pulmonary autograft transplantation in the surgical treatment of aortic valve disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Aortic root replacement with pulmonary autograft (Ross procedure) has the advantages of good haemodynamics and growth potential without the need for anticoagulation. In this study, we reviewed our experience of the Ross procedure for patients with aortic valve disease.</p><p><b>METHODS</b>From October 1994 to January 2005, 42 Ross procedures were performed in our centre. There were 30 males and 12 females. The mean age was 28 +/- 15 years (range, 5-56 years). Congenital heart disease (CHD) with aortic valve stenosis (AS) and/or aortic valve insufficiency (AI) in 40 cases including one associated with ventricular septal defect (VSD), degenerated aortic valve disease with AS in 1 and subacutive bacterial endocarditis (SBE) with AI in 1 were studied. The diagnosis was made by ultracardiography (UCG) in all patients. The mean aortic valve annulus diameter (AVD) was (2.45 +/- 0.31) cm and pulmonary valve annulus diameter (MPVD) was (2.34 +/- 0.21) cm. All patients had normal pulmonary valves. The New York Heart Association (NYHA) function class was II in 36 cases and III in 6 cases. The operation was performed under moderate hypothermic cardiopulmonary bypass (CPB) with aortic root replacement using pulmonary autograft and pulmonary valve replacement with a homograft.</p><p><b>RESULTS</b>There was no early hospital mortality. Postoperative UCG showed normal aortic valve function in all our patients. The mean gradient across the aortic valve was (6.11 +/- 0.12) mmHg. The left ventricular diastole diameter (LVDD) decreased significantly from (62 +/- 5) mm to (56 +/- 3) mm (P < 0.001). The mean postoperative left ventricular ejective fraction (LVEF) was 0.49 +/- 0.23. All patients were in NYHA class I-II. Follow-up was completed in 38 cases for a mean period of 3.2 years (range 1-10 years). All survivors were in NYHA class I with normal neo-aortic and pulmonary valve function. One patient died after secondary operation due to homograft fungal endocarditis 1 year after the Ross procedure. The cause of death was uncontrolled bleeding. Another patient suffered from cardiogenic shock and was on extracorporeal membrane oxygenation (ECMO) for 10 days postoperatively. This patient was subsequently self-discharged from hospital due to financial issues and he was excluded from follow-up.</p><p><b>CONCLUSION</b>The Ross procedure is an excellent technique to treat aortic valve disease. Our data show that it can be performed safely with good early and mid-term clinical outcomes.</p>