Objective To study some biological properties of mouse bone marrow stromal cells(MSCs)and their ability to differentiate endothelial cells and surviving ability in ischemic tissue,and provide an experimental foundation for applying MSCs to ischemic repair.Methods After the tibias and femurs were dissected from 5-to 6-week-old mice from Jan.2005 to Nov.2005,the marrow was flushed out with ice-cold DMEM/F12 medium.The mononuclear cells of the marrow were obtained with density gradient centrifugation and the plated and cultured in DMEM/F12 medium.The cultured cells in vitro were induced with endothelial cell growth supplement.The ability of MSCs was also was examined in ischemic tissue.Results The adherent fibroblast-shaped cells approached confluence in single layer 12~16 d after plating.The cultured MSCs in vitro differentiated into endothelium.Numerous scattered DAPI-labeled cells were found in the specimen seven days after implantation,and hematoxylin and eosin staining of the adjacent section showed concordance between dense hematoxylin staining and presence of DAPI epifluorescence,and there was no obvious inflammatory response.Conclusion The subcultured MSCs possess potential to differentiate into endothelial cells.MSCs show stable growth in vitro,easy survival in the subculture and rapid proliferation in present culture condition.MSCs may be used in therapy for myocardium ischemia.

Adult ; Aged ; Aged, 80 and over ; Asbestos ; adverse effects ; Environmental Exposure ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Pleural Diseases ; chemically induced ; diagnostic imaging ; Tomography, X-Ray Computed

Objective To discuss the synergistic mechanism of compatibility of Tripterygium Wilfordii and Astragalus,and to determine wilforlide A before and after compatibility.Methods To research Tripterygium Wilfordii single medicinal materials,Tripterygium Wilfordii compatibility of Astragalus and Tripterygium Wilfordii reference substance using the HPLC method,so as to provide further verification by the change of wilforlide A.Results The content of wilforlide A was 0.00 324% by measuring Tripterygium Wilfordii single medicinal materials.The content of wilforlide A was 0.00 184% by measuring Tripterygium Wilfordii compatibility of Astragalus.Conclusion The content of wilforlide A was decreased before and after compatibility and toxicity can be reduced.The synergistic mech-anism of the compatibility of Tripterygium Wilfordii and Astragalus is conteract the toxicity of another drugin posse.

Objective:To observe influence of simvastatin on p 27 protein (cyclin‐dependent kinase inhibitor ) expres‐sions of vascular smooth muscle cells (SMC) and endothelial cells (EC) in rats for screening new generation coating drugs of eluting stents .Methods :Primary aortic SMC and EC of rat were isolated and cultured by methods of adher‐ent and enzymatic digestion respectively .Which were inoculated on fibronectin -coated culture plates .α smooth muscle actin immunofluorescence staining was used to identify SMC ,and von Willebrand factor (vWF) immunofluo‐rescence staining was used to identify EC .SMC and EC were cultured for 24h with different concentrations of simv‐astatin (0.01 ,0.1 ,1 and 10 μmol/L) ,then Western blot was used to measure p27 protein expression .Results:Compared with blank control group ,0.01μmol/L simvastatin had no significant influence on p 27 protein expression of SMC ,but 0.1 ,1 and 10 μmol/L simvastatin significantly raised p27 protein expression of SMC [ (0.53 ± 0.08) vs .(0.86 ± 0.05) ,(1.20 ± 0.05) ,(1.60 ± 0.04)] , P< 0.01 all .Besides ,different concentrations of simvastatin had no significant influence on p27 protein expression of EC , P> 0.05 ,indicating that simvastatin only dose‐de‐pendently promoted p27 protein expression of SMC .Conclusion:Simvastatin dose -dependently promotes p27 pro‐tein expression of vascular smooth muscle cells without affecting p 27 protein expression of endothelial cells .So local application of simvastatin may inhibit restenosis and promote reendothelialization of injured vessels .

Objective To investigate the risk factors and early diagnosis of the severe thrombocytopenia complicating transcatheter ccclusion of patent ductus arteriosus ( PDA ) . Methods Between February, 2011 and May, 2015, 80 patients with patent ductus arteriosus underwent percutaneous intervention occlusion were studied. Results Average age were ( 17. 5 ± 17. 1 ) years, 63 were females (78. 8%), mean weight were (35. 6 ± 20. 2)kg (from 6. 0 to 75. 0 kg), mean body surface area (BSA) were (1. 09 ±0. 44) m2(from 0. 32 to 1. 91 m2). A bolus of heparin calcium (80 U/kg) was administered by intravenous injection. The mean diameters of patent ductus arteriosus were 4 mm (from 2 to 18 mm), and the mean diameters of occluders were 12 mm (from 6 to 30 mm). 14 patients were found to have severe thrombocytopenia (PLT count﹤100 × 109/L). The reduction rate of platelet in 12 of 14 patients was more than 19%. The diameters of all occluders were equal to or more than 14 mm, the mean diameters of patent ductus arteriosus were 10 mm ( from 6 to 18 mm) and the mean diameters of occluders were 18 mm ( from 14 to 30 mm). All the 14 patients started to present progressive decrease in PLT count since the second day post procedure. Taking the selected occluder diameter greater than 14 mm as cut-off points in diagnosis of severe thrombocytopenia, the sensitivity was 100%, specificity was 68%, the positive predictive value was 40%, and the negative predictive value was 100%. Combined with the postprocedural second day complete blood count analysis and the platelet count decreased by 10% as cut-off points in diagnosis of severe thrombocytopenia patients, the sensitivity was 93%, specificity was 67%, the positive predictive value was 65%, the negative predictive value was 93% . If taking the platelet count decreased by 7% on second day as cut-off points in diagnosis of severe thrombocytopenia patients, the sensitivity was 100%, specificity was 57%, the positive predictive value was 61%, the negative predictive value was 100% . Logistic regression analysis discovered that risk factors of severe thrombocytopenia after PDA are procedural platelet count and occluder diameter. Conclusions The risk factors of severe thrombocytopenia complicating transcatheter ccclusion of patent ductus arteriosus were the procedural reduction of platelet count and big occluder diameter. Patients with PDA who were inplanted with occluders equal to or bigger than 14 mm should retest the numbers of platelet on the second day after procedure and retest on third day if the numbers reduce on the second day, which may help in the prediction of severe thrombocytopenia.

Objective:To observe influence of simvastatin on differentiation ,proliferation ,migration and adhesion of marrow-derived smooth muscle progenitor cells (SPCs) and screen coated eluting stent drugs of new generation . Methods :The mononuclear cells (MNCs) were isolated from rat marrow by density gradient centrifugation method , and then plated on fibronectin-coated culture dishes ,after culture 8d ,marrow-derived SPCs were identified by α-smooth muscle actin (α-SMA) immunofluorescent staining and counted under inverted fluorescence microscope .The MNCs and adhesion cells were treated with simvastatin (0.01~10 μmol/L) respectively for 8 d and 24h .SPCs pro-liferation ,migration and adhesion were observed by Tritium thymidine (3 H-TdR) intake method ,modified Boyden chamber assay and adhesion assay .Results:Compared with control group (no simvastatin intervention ) ,0.01μmol/L simvastatin significantly inhibited the MNCs differentiation towards SPCs [ (85 ± 4) vs .(79 ± 5)] ,proliferation [ (4070 ± 184) vs .(3833 ± 126)] ,migration [ (44 ± 3) vs .(39 ± 3)] and adhesion of SPCs [ (59 ± 5) vs .(52 ± 4)] , P<0.05 all ,and number of SPCs significantly reduced along with simvastatin concentration increased (P<0.01) . Conclusion:Simvastatin could inhibit the differentiation ,proliferation ,migration and adhesion of marrow-derived smooth muscle progenitor cells .

Objective: To study the expressions of endothelin-1 in primary cultured atrial myocyte model at early stages of atrial fibrillation by rapid pacing. Methods: The primary rat atrial myocytes were cultured, in which a rapid paced cell model was established. The polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the messenger ribonucleic acid (mRNA) expression and secretion of endothelin-1 on 3 h, 6 h, 12 h and 24 h after rapid pacing. Results: Compared with before pacing, there were continuous and significant increase in expression of ET-1mRNA [ (0.31±0.02) vs. (0.52±0.07), (0.62±0.09), (0.75±0.05) ] and secretion of endothelin-1 [ (3.4±0.8) ng/L vs. (6.0±1.4) ng/L, (8.3±1.5) ng/L, (11.2±2.1) ng/L] on 6 h, 12 h and 24 h after rapid pacing, P<0.01 all. Conclusion 　 Expressions of ET-1mRNA and secretion of endothelin-1 significantly increase depend on time in early rapid pacing atrial myocytes, indicating that endothelin-1 participates in atrial pathological remodeling during atrial fibrillation.

Objective to investigate the relationship between the size of ostium secondary atrial septal defect (ASD) and the pulmonary arterial pressure (PAP) in children less than 5 years of age. Methods During the period from April 2000 to January 2011, a total of 189 child patients less than 5 years of age with ostium secondary ASD were admitted to General Hospital of Shenyang Military Command. Under general anaesthesia with ketamine cardiac catheterization was performed, PAP was measured, and percutaneous occlusion of ASD was carried out in all patients. The clinical indexes, including sex, age, body height, body weight, body surface area, diameter of ASD defect, heart- to- thorax ratio, the systolic, diastolic and mean pressure of the pulmonary artery, etc. were determined. The patients were followed up for one year and postoperative cardiac ultrasonography was performed to check the results. The patients were divided into groups according to the defect size. Results The 189 patients consisted of 77 males and 112 females with a male-to-female ratio of 1 ∶ 1.5. The mean age was (4.1 ± 0.9) years old, ranging from 2 to 5 years old. The mean weight was (17.2 ± 3.6) kg, ranging from 10.0 to 30.0 kg. The mean height was (104.9 ± 9.2) cm, ranging from 77 to 135 cm. The mean body surface area (BSA) was (0.71 ± 0.10) m2, ranging from 0.46 to 1.02 m2. The mean size of ASD was (12.6 ± 4.8) mm, ranging from 5 to 29 mm. The mean size of ASD, which was modified by BSA, was (18.0 ± 7.0) mm/m2, ranging from 5.3 to 38.9 mm/m2. The mean systolic PAP was (41.1 ± 8.9) mmHg with a range of 15 - 67 mmHg. The mean diastolic PAP was (16.8 ± 6.5) mmHg with a range of 3 - 45 mmHg. The mean PAP was (24.9 ± 6.7) mmHg with a range of 12 - 48 mmHg. One hundred and fifty- nine patients (89.4%) had pulmonary arterial hypertension (PAH) which was determined by right heart catheterization, but no patient showed PAH when the pulmonary arterial pressure was measured by echocardiography before the procedure as well as 1, 3, 6, 12 months after the procedure. No definite correlation existed between the size of ASD and the pulmonary artery pressure (P > 0.05). Conclusion Pulmonary artery pressure measured by right heart catheterization has no definite correlation with the size of ASD in children less than 5 years of age. Pulmonary artery pressure obtained from right heart catheterization is higher than that determined by cardiac ultrasonography, which may be caused by the effect of ketamine when general anaesthesia is used in performing right heart catheterization.

Objective To investigate the different influences of simvastatin on proliferation of rat smooth muscle progenitor cells(SPCs) versus endothelial progenitor cells (EPCs) and identify the compounds that differentially inhibit SPCs and EPCs proliferation for clinical usefulness. Methods Total mononuclear cells (MNCs) were isolated from bone marrow of rats by Fieoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. SPCs outgrew from the culture of MNCs in the presence of platelet-derived growth factor-BB and basic fibroblast growth factor, whereas EPCs were obtained in the presence of vascular endothelial growth factor. SPCs were identified as adherent cells positive for α-smooth muscle actin (α-SMA) by indirect immunofluoreseent staining. EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining. SPCs and EPCs were stimulated by simvastatin (0.01～10.00 μmol/L) or vehicle control for the respective time points (6 h, 12 h, 24 h and 48 h). SPCs and EPCs proliferation were assayed with 3H-TdR incorporation and manual counting respectively. Results Simvastatin obviously inhibited SPCs proliferation. At the concentration of 0. 01 μmol/L for 12 h,simvastatin significantly reduced the number of SPCs by (5.8±3.1)% compared with control group (P<0.05). Simvastatin significantly stimulated EPCs proliferation, which was dose- and time dependent and reached maximum at 1 μmol/L after 24 hours (2.0±0.1 fold increase, P<0.01).Conclusions Simvastatin displays different effects on SPCs (inhibited) and EPCs (promoted)proliferation. Local application of simvastatin may inhibit arterial restenosis and promote reendothelialization of injured vessels.

The fruit of Pandanus tectorius (PTF) has a long history of use as a folk medicine to treat hyperlipidemia in Hainan province, South China. Our previous studies have shown that the n-butanol extract of PTF is rich in caffeoylquinic acids and has an adequate therapeutic effect on dyslipidemic animals induced by high-fat diet. In this work, seven caffeoylquinic acids isolated from PTF were screened for the lipid-lowering activity in HepG2 hepatoma cells. Oil-Red O staining, microscopy and intracellular triglyceride (TG) and total cholesterol (TC) quantification showed that 3-O-caffeoylquinic acid (3-CQA), 3, 5-di-O-caffeoylquinic acid (3,5-CQA), and 3,4,5-tri-O-caffeoylquinic acid (3,4,5-CQA) significantly inhibited lipid accumulation induced by oleic acid and decreased intracellular levels of TC and TG in a dose-dependent manner. These three caffeoylquinic acids showed no significant cytotoxicity at concentrations of 1 -50 μmol x L(-1) as determined by MTT assay. Realtime quantitative PCR revealed that 3-CQA and 3, 5-CQA significantly increased the expression of lipid oxidation-related genes PPARα, CPT-1 and ACOX1 while 3-CQA, 3, 5-CQA and 3,4,5-CQA decreased the expression of lipogenic genes SREBP-1c, SREBP-2, HMGR, ACC, FAS. Overall, 3-CQA, 3, 5-CQA and 3, 4, 5-CQA may be the principal hypolipidemic components in PTF which can decrease intracellular lipid accumulation through up-regulating the expression of lipid oxidative genes and down-regulating the expression of lipogenic genes.
Carcinoma, Hepatocellular ; metabolism ; China ; Cholesterol ; metabolism ; Gene Expression Regulation ; Hep G2 Cells ; Humans ; Lipid Metabolism ; Liver Neoplasms ; metabolism ; Oleic Acid ; Pandanaceae ; chemistry ; Quinic Acid ; analogs & derivatives ; chemistry ; Sterol Regulatory Element Binding Protein 1 ; Triglycerides ; metabolism