ObjectiveTo study the relation between white blood cell(WBC) count and left ventricular (LV) remodeling after emergency percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI).MethodsA total of 117 ST segment elevation AMI patients having underwent emergency PCI were enrolled.WBC count,cardiac troponin Ⅰ (cTnI),high-sensitivity C-reactive protein (hs-CRP) and N-terminal pro-B-type natriuretic peptide(NT-proBNP) were obtained at admission before PCI.According to the WBC count level,patients were divided into normal WBC group(WBC count ≤ 10 ×109/L,60 cases) and elevated WBC group(WBC count＞ 10 × 109/L,57 cases).Two-dimensional echocardiography was applied after PCI.The relation between WBC count and LV remodeling prognosis including LV ejection fraction(LVEF),LV end diastolic diameter(LVEDD) and LV aneurysm were compared after AMI.ResultsAdmission NT-proBNP,hs-CRP and cTnI peak in elevated WBC group were higher than those in normal WBC group [ (2408.83 ± 3173.39) pg/L vs.(713.11 ± 636.82) pg/L,(39.64 ± 59.51) mg/L vs.(11.23 ± 14.14) mg/L,(107.76 ± 107.71) pg/L vs.(62.23 ± 87.79) pg/L,P ＜0.05].Admission WBC count was positively correlated with LVEDD and negatively correlated with LVEF(P ＜0.01 ).Patients with LV aneurysm had higher WBC count than those without LV aneurysm[ ( 12.59 ± 5.22) × 109/L vs. (9.27 ± 2.60) × 109/L,P =0.001 ].Multivariate analyses showed that admission WBC count ≥ 10.5 × 109/L was an independent determinant of LV aneurysm(OR =22.5,95％ CI:2.69-187.83,P ＜ 0.01 ),and this cut-off value yielded sensitivity of 76.9％ and specificity of 69.7％ respectively.Conclusion Admission WBC count may be considered as a prognostic biological tag in the prediction of the development of LV remodeling after emergency PCI in patients with AMI.

Objective To investigate the effect of iodine excess on thyroid follicle epithelial ultrstructure and the relationship between thyroid iniury and autoimmune thyroiditis. Methods NOD.H-2h4 mice and Kunming mice were randomly divided into four groups receiving plain water,5 fold,10 fold,and 100 fold excessive iodine water.4,8 and 24 weeks after receiving iodine water,the mice were killed.After fixation with osmic acid and dual staining with uranyl chloride and citrate lead,thyroid gland ultrstructure was examined with electron microscopy.Resuits Iodine treated NOD.H-2h4 mice exhibited marked accumulation of peroxisome and secondary lysosomes,apoptosis and necrosis of thyroid epithelial cell.damage of thyroid follicles and lymphocvtic infiltration.The observed changes induced by iodine were in a dose dependent way.Conclusion The oxidative iniury on the thyroid epithelial cells induced by iodine excess might be the prerequisite for the creation of autoimmune thyroiditis.

Objective To evaluate the cause of evaluated postage-specific antigen(PSA)in patients performed transrectal uhrasonography(TRUSG)guided prostate biopsy beeause of high PSA levels.Methods In a retrospective study 504 prostate biopsies performed between January 1998 and December 2001 were evaluated and the levels of serum PSA were determined in samples obtained immediately before sextant biopsy was performed.All patients underwent 6 or 13 cote primary prostate needle biopsies.Results 185 prostate cancer,109 NIH-Ⅳ prostatitis and 210 patients with benign prostate hyperplasia(BPH)were identified.The difference in free-PSA,total-PSA,f/t-PSA levels between prostate cancer,BPH and NIH-Ⅳ prostatitis was significant(P＜0.05).No significant difference was found in age,transrectal ultrasonography and digital rectal examination.In multivariate analysis,free-PSA,total-PSA,f/t-PSA was the significant predictors of histology in prostate cancer(P＜0.05).A significant correlation was found between the serum total and free PSA levels and the grade and stage of prostate cancer(P＜0.05).Preoperative variables predictors of histology in BPH were TPSA and FPSA(P＜0.05).In multivariate analysis,TPSA was the only significant predictors of histology in BPH(P＜0.01).The best cutoff value was constructed to differ pathology type in prostate diseases:tPSA≥4 ng/ml,fPSA≥0.85 ng/ml and f/t-PSA≤0.16(P＜0.05).Conclusions High serum PSA levels may correlate with asymptomatic inflammatory prostatitis,prostate cancer and BPH.The factors contributing to elevated serum PSA concentrations include cell proliferating,glandular epithelial disrupt.

Objective To discuss the relationship between asymptomatic prostatic inflammation (NIH category Ⅳ prostatitis)and serum prostate specific antigen. Methods In a retrospective study,245 prostate biopsies with benign pathological results from January 1998 to January 2000 were reviewed and the corresponding serum PSA before biopsy were analyzed.All patients were taken 6 or 13 cores prostate biopsy. Results One hundred and twenty-seven NIH-Ⅳ prostatitis and 118 patients with benign prostatic hyperplasia(BPH)were identified.The difference in free-PSA,totalPSA,f/t-PSA levels between BPH and NIH-Ⅳ prostatitis was significant(P＜0.05).In multivariate analysis,free-PSA,total-PSA,f/t-PSA was the significant predictors of histology in NIH-Ⅳ prostatitis(P＜0.05).The best prediction factors were constructed to predict pathology type in NIH-Ⅳ prostatitis:TPSA≥4 ng/ml,fPSA≥0.85 ng/ml and f/t-PSA≤0.16(P＜0.05). Conclusions Asymptomatic inflammation of the prostate was one of the confounding factors in patients with an elevated PSA.In the diagnosis of prostate diseases,it should be taken into account prostatitis might elevate the level of PSA.

OBJECTIVETo investigate the effects of endothelial microvesicles (EMVs) induced by calcium ionophore A23187 on H9c2 cardiomyocytes.

METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with 10 micromol/L A23187 for 30 min. EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. EMVs were characterized using 1 and 2 microm latex beads and anti-PE-CD144 antibody by flow cytometry. For functional research, EMVs at different concentrations were cocultured with H9c2 cardiomyocytes for 6 h. Cell viability of H9c2 cells and the activity of LDH leaked from H9c2 cells were tested by colorimetry. Moreover, apoptosis of H9c2 cells was observed through Hoechst 33258 staining and tested by FITC-Annexin V/PI double staining.

RESULTSEMVs were induced by A23187 on HUVECs, and isolated by ultracentrifugation. We identified the membrane vesicles (< 1 microm) induced by A23187 were CD144 positive. In addition, the EMVs could significantly reduce the viability of H9c2 cells, and increase LDH leakage from H9c2 cells in a dose dependent manner (P < 0.05). Condensed nuclei could be observed with the increasing concentrations of EMVs through Hoechst 33258 staining. Furthermore, increased apoptosis rates of H9c2 cells could be assessed through FITC-Annexin V/PI double staining by flow cytometry.

CONCLUSIONMicrovesicles could be released from HUVECs after induced by A23187 through calcium influx, and these EMVs exerted a pro-apoptotic effect on H9c2 cells by induction of apoptosis.

Annexin A5 ; Apoptosis ; Calcimycin ; pharmacology ; Calcium ; metabolism ; Cell Line ; Cell Membrane ; drug effects ; Coculture Techniques ; Flow Cytometry ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; Human Umbilical Vein Endothelial Cells ; Humans ; Myocytes, Cardiac ; drug effects ; Staining and Labeling

OBJECTIVETo construct a eukaryotic expression vector of transmembrane-4-L-six-family-1 (TM4SF1) gene and study its effect on the migration and invasion of colorectal cancer cells.

METHODSA pair of specific primers of TM4SF1 gene (GenBank: BC034145.1) was used to acquire the open reading frame of TM4SF1 by RT-PCR. The amplified sequence was ligated to a PEZ-M29 vector, which, after identification, was transiently transfected in colorectal cancer cell lines HCT116 and DLD1. Western blotting and immunocytochemistry were used to analyze the transfection efficiency, and scratch and Transwell tests were performed to analyze the changes in the migration and invasion of HCT116 and DLD1 cells after transfection.

RESULTSCell scratch and Transwell assays revealed that transfection with the recombinant plasmid, PEZ-M29/TM4SF1, caused up-regulated expression of TM4SF1 and promoted the migration and invasion of HCT116 and DLD1 cells.

CONCLUSIONOur results demonstrated that TM4SF1 is closely related to the invasion and metastasis of colorectal cancer cells in vitro.

Antigens, Surface ; biosynthesis ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Genetic Vectors ; HCT116 Cells ; Humans ; Neoplasm Proteins ; biosynthesis ; Transfection

Objective To construct a eukaryotic expression vector of transmembrane-4-L-six-family-1 (TM4SF1) gene and study its effect on the migration and invasion of colorectal cancer cells. Methods A pair of specific primers of TM4SF1 gene (GenBank: BC034145.1) was used to acquire the open reading frame of TM4SF1 by RT-PCR. The amplified sequence was ligated to a PEZ-M29 vector, which, after identification, was transiently transfected in colorectal cancer cell lines HCT116 and DLD1. Western blotting and immunocytochemistry were used to analyze the transfection efficiency, and scratch and Transwell tests were performed to analyze the changes in the migration and invasion of HCT116 and DLD1 cells after transfection. Results Cell scratch and Transwell assays revealed that transfection with the recombinant plasmid, PEZ-M29/TM4SF1, caused up-regulated expression of TM4SF1 and promoted the migration and invasion of HCT116 and DLD1 cells. Conclusion Our results demonstrated that TM4SF1 is closely related to the invasion and metastasis of colorectal cancer cells in vitro.

Objective To construct a eukaryotic expression vector of transmembrane-4-L-six-family-1 (TM4SF1) gene and study its effect on the migration and invasion of colorectal cancer cells. Methods A pair of specific primers of TM4SF1 gene (GenBank: BC034145.1) was used to acquire the open reading frame of TM4SF1 by RT-PCR. The amplified sequence was ligated to a PEZ-M29 vector, which, after identification, was transiently transfected in colorectal cancer cell lines HCT116 and DLD1. Western blotting and immunocytochemistry were used to analyze the transfection efficiency, and scratch and Transwell tests were performed to analyze the changes in the migration and invasion of HCT116 and DLD1 cells after transfection. Results Cell scratch and Transwell assays revealed that transfection with the recombinant plasmid, PEZ-M29/TM4SF1, caused up-regulated expression of TM4SF1 and promoted the migration and invasion of HCT116 and DLD1 cells. Conclusion Our results demonstrated that TM4SF1 is closely related to the invasion and metastasis of colorectal cancer cells in vitro.

OBJECTIVETo establish a flow cytometric method to detect the alteration of phenotypes and concentration of circulating microvesicles (MVs) from myocardial ischemic preconditioning (IPC) treated rats (IPC-MVs), and to investigate the effects of IPC-MVs on ischemia/reperfusion (I/R) injury in rats.

METHODSMyocardial IPC was elicited by three.cycles of 5-min ischemia and 5-min reperfusion of the left anterior descending (LAD) coronary artery. Platelet-free plasma (PFP) was isolated through two steps of centrifugation at room temperature from the peripheral blood, and IPC-MVs were isolated by ultracentrifugation from PFR PFP was incubated with anti-CD61, anti-CD144, anti-CD45 and anti-Erythroid Cells, and added 1, 2 µm latex beads to calibrate and absolutely count by flow cytometry. For functional research, I/R injury was induced by 30-min ischemia and 120-min reperfusion of LAD. IPC-MVs 7 mg/kg were infused via the femoral vein in myocardial I/R injured rats. Mean arterial blood pressure (MAP), heart rate (HR) and ST-segment of electro-cardiogram (ECG) were monitored throughout the experiment. Changes of myocardial morphology were observed after hematoxylin-eosin (HE) staining. The activity of plasma lactate dehydrogenase (LDH) was tested by Microplate Reader. Myocardial infarct size was measured by TTC staining.

RESULTSTotal IPC-MVs and different phenotypes, including platelet-derived MVs (PMVs), endothelial cell-derived MVs (EMVs), leucocyte-derived MVs (LMVs) and erythrocyte-derived MVs (RMVs) were all isolated which were identified membrane vesicles (<1 Vm) with corresponding antibody positive. The numbers of PMVs, EMVs and RMVs were significantly increased in circulation of IPC treated rats (P<0.05, respectively). In addition, at the end of 120-min reperfusion in I/R injured rats, IPC-MVs markedly increased HR (P<0.01), decreased ST-segment and LDH activity (P < 0.05, P < 0.01). The damage of myocardium was obviously alleviated and myocardial infarct size was significantly lowered after IPC-MVs treatment (P < 0.01).

CONCLUSIONThe method of flow cytometry was successfully established to detect the phenotypes and concentration alteration of IPC-MVs, including PMVs, EMVs, LMVs and RMVs. Furthermore, circulating IPC-MVs protected myocardium against I/R injury in rats.

Animals ; Cell-Derived Microparticles ; metabolism ; Coronary Vessels ; pathology ; Flow Cytometry ; Heart Rate ; Ischemic Preconditioning, Myocardial ; Myocardial Infarction ; physiopathology ; Myocardial Reperfusion Injury ; physiopathology ; Myocardium ; pathology ; Phenotype ; Rats

OBJECTIVETo establish an improved three-dimension (3D) and serum-free approach to differentiate human embryonic stem cells (hESCs) into endothelial cells, and detect the endothelial functions of the obtained cells.

METHODSWe cultured undifferentiated H9 human embryonic stem cell line in low-adhesion dishes to form embryonic bodies (EBs). After 12 days, EBs were harvested, re-suspended into rat tail collagen type I, and put into the incubator (37℃). After 30 minutes, EGM-2 culture medium was added to the solidified collagen, and the EBs were cultured for another 3 days to form embryonic body-sproutings (EB-sproutings). EB-sproutings were digested with 0.25% collagenase I and 0.56 U/ml Liberase Blendzyme for 20 minutes respectively, and the CD31(+) cells were sorted by FACS. The endothelial functions were tested by Dil-ac-LDL uptake assay and tube formation assay.

RESULTSThis approach raised the efficiency of endothelial differentiation to 18%, and also avoided the contamination with animal materials. The obtained hESC-derived endothelial cells (hESC-ECs) had the similar pattern of surface biomarkers as human umbilical vein endothelial cells (HUVECs), and their endothelial functions were confirmed by the uptake of Dil-ac-LDL and the tube formation on Matrigel.

CONCLUSIONSThe improved 3D approach can enhance the efficiency of differentiation from hESCs into endothelial cells. Furthermore, serum free differentiation system may be applied in future hESC-based therapies for various ischemic diseases.

Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Line ; Collagen Type I ; Culture Media ; Embryonic Stem Cells ; cytology ; Endothelial Cells ; cytology ; Humans