| Objective: To extract, culture and identify rat bone marrow-derived EPCs in vitro,labeled cells with5-BrdU.Methods: The SD rat femur and tibia bone marrow mononuclear cells (MNCs) wereisolated and extracted by Percoll density gradient centrifugation and speed differenceadhesion culture. MNCs were cultured with administration of basic fibroblast growthfactor (bFGF), vascular endothelialcell growth factor (VEGF) and epidermal growthfactor (EGF) for14days. The morphological features were further observed withoptical microscope. Immunocytochemistry was used to identify their surface antigenexpression. Double fluorescence staining was used to detect Di-labeled Acetyl lowdensity lipoprotein (Di-acLDL) and FITC-labeled Ulex europaeus agglutinin (UEA-1).Identified EPCs were incubated with5-BrdU. EPCs were incubated with5-BrdU atdifferent concentrations, cells proliferation was assayed by MTT. EPCs Labeled with5-BrdU were for subsequent use.Results: Adherent cells grew showing an initial cobblestone, and then a spindle-like arrangement style. CD34, CDl33, Flk-1(VEGFR) and vWF (VIII of factor) werepositively expressed at different stages in the adherent cells. The EPCs uptookDi-acLDL and bound FITC-UEA-1.5-BrdU can be used to lable EPCs,did not affectcells proliferation.Conclusions: Percoll density gradient centrifugation and speed difference cultivationcombined withVEGF, bFGF, and EGF could obtain highly purified EPCs, and thesecells showed some of the features of mature ECs.5-BrdU can be used to lable EPCs. Objective: To prepare anti-myosin light chain/CD34bispecific antibody (BsAb) bychemical heteroconjugation, and to assemble them on the surface of EPCs.Methods: EPCs were extracted, isolated and cultured as described above. BsAb andFITC-BsAb were generated by chemical heteroconjugation. EPCs were mixed withFITC-BsAb at10ng,20ng,50ng and100ng, the binding activity was detected byfluorescence and flow cytomety to screen out the optimal concentration of the antibody.EPCs were assembled with these BsAbs for further purpose.Results: BsAb was successfully generated and assembled on the surface of EPCs. Themost optimal concentration was50ng/106cells.Conclusions: BsAb was successfully generated and further assembled on the surfaceof EPCs. Objective: To study the feasibility of ultrasound microbubble destruction promotingBsAb-labeled EPCs to transplant to the ischemic myocardium.Methods: BsAb-labeled EPCs were obtained as previously discribed.426-week,healthy SD rats were used as acute myocardial infarction model. They were divided asfollowing:1. Normal;2. MI;3. Antibody;4. US and MB;5. EPCs;6EPCs andantibody;7. EPCs, US, MB and antibody. Rats were treated with ultrasound andtransplanation8hours after modeling. The last group was injected1ml BsAb-labeled ofEPCs, and was further injected with lipid ultrasound microbubble solution1ml. Theprecordium of the rats were irradiation. After30days, the myocardial tissues werestained with5-BrdU to detect the migration and distribution of EPCs.Echocardiography was used to detect the heart function. The function were detected byleft ventricular ejection fraction (LVEF) and left ventricular fractional shortening (FS).VIIIwas detected by immunochemistry after30days. Real time PCR and western blotwere equipped to detect the expression of VEGF and SDF-1.Results: Group7(EPCs, UM and antibody) was stained with more5-BrdUsignificantly. Group7showed high LVEF and FS compared with MI groups. Group7had more positive capillaries. The mRNA and protein level of VEGF and SDF-1ingroup7rats increased compared with MI groups through real time PCR and Westernblot. There was no difference between group7and Normal group of the mRNA andprotein level of VEGF and SDF-1through real time PCR and Western blot.Conclusions: Combined with UM, BsAb could promote the target-directed migrationof EPCs and capillaries genesis. The work provided new strategy for the treatment of myocardial infarction. Objective: to clarify the association between FTO polymorphism and cancer risk.Methods: Published literature from PubMed and Embase databases had been retrieved.Pooled odds ratio (OR) with95%confidence interval (CI) was calculated byfixed-effects model.13studies involving16,277cases and31,153controls had beenidentified.Results: FTO polymorphism was not significantly associated with the increased riskof cancer (OR=1.01,95%CI0.98–1.04), except that a statistically significantassociation with pancreatic cancer (OR=1.10,95%CI1.03–1.19). No publication biaswas detected (Begg's test: P=0.760; Egger's test: P=0.553).Conclusions: Our meta-analysis implicated that no association between FTOpolymorphism and the increased risk of cancer existed, although it was marginallyassociated with pancreatic cancer. However, the conclusion should be made withcaution for most included studies had not take BMI/obesity into account. |
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