Objective： To investigate the effect of oxLDL and VitE on the expression of CD40 and CD40 ligand(CD40L) in cultured human monoc ytes. Methods： The expression of CD40 and CD40L on monocytes su rface were measured by indirect immunorescence technique in combination with flo w cytometry. Results： Low concentration of oxLDL（≤200 μg/L） significantly increased the expression of CD40 and CD40L in a dose and time dep endent manner. High concentration (>200 μg/L）of oxLDL markedly reduced the exp ression of CD40 and CD40L. When VitE was added, it significantly reduced the ex pression of CD40 and CD40L on monocytes surface induced by oxLDL in a dose-depe ndent manner. Conclusion：It is an important mechanism that the high expression of CD40 and CD40L induced by oxLDL may be contributed to the for mation of atherosclerosis. Antioxidan VitE can partially inhibit the high expres sion of CD40 and CD40L on monocytes surface induced by oxLDL.

Objective： To investigate the effect of c ytokines (IFN-γ，TNF and IL-1) on the expression of CD40 and CD40 ligand (CD4 0L) in monocytes/macrophages. Methods： The mRNA expression of C D40 and CD40L was measured by RT-PCR and the CD40，CD40L expression on the mono cytes/macrophages were detected by flow cytometric analysis. Results： IFN-γ，TNF and IL-1 could not only significantly up-regulate the mRNA levels of CD40 and CD40L in cultured monocytes/macrophages, but also increase t he expression of CD40 and CD40L. Antioxidant VitE could reduce the expression o f CD40 and CD40L induced by IFN-γ，TNF and IL-1. Conclusion： IFN-γ，TNF and IL-1 can stimulate high expression of CD40 and CD40L . Antio xidant VitE can partially inhibit the expression of CD40 and CD40L induced by cy tokines in cultured monocytes/macrophages.

OBJECTIVETo explore if reduced number of circulating endothelial progenitor cells (EPCs) is a risk factor for patients with coronary slow flow (CSF).

METHODSThirty patients with CSF and 30 age and gender matched control subjects with normal coronary angiography were included in the study. Mononuclear cells were isolated from peripheral blood by Ficoll density gradient centrifugation and plated on fibronectin-coated culture dishes. EPCs were characterized as adherent cells double positive for DiI-AcLDL-uptake and lectin-binding by converted fluorescence microscope (×200).

RESULTSSmoking, diabetes mellitus, hypertension and the levels of plasma lipoprotein profile were similar between the two groups (all P > 0.05). The number of EPCs was significantly lower in patients with CSF compared with control subjects (35.7 ± 5.9 vs.53.2 ± 5.9, P < 0.01). TIMI frame counts was correlated with circulating EPCs number (OR = 0.424, 95%CI 0.358 - 0.621, P < 0.01) and not associated with gender, age, smoking, diabetes mellitus, hypertension and the levels of plasma lipoprotein profile.

CONCLUSIONDecreased circulating EPCs is an independent risk factor for CSF.

Blood Circulation ; Blood Flow Velocity ; Case-Control Studies ; Cell Count ; Cells, Cultured ; Coronary Angiography ; Coronary Vessels ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Risk Factors ; Stem Cells ; cytology

OBJECTIVETo investigate rapid atrial pacing (RAP) induced atrial ultrastructural changes and mRNA and protein expression changes of L-type calcium channel subunits and potassium channel Kv4.3 in a rabbit model.

METHODSThirty-six rabbits were electrically paced at a frequency of 600 beats/min for durations ranging from 0 - 48 h via bipolar endocardial leads through surgical techniques. Ultrastructural changes of the atrium were observed through a transmission electron microscope (TEM), L-type calcium channel subunits and potassium channel Kv4.3 expressions at mRNA and protein levels were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.

RESULTSAtrial ultrastructure changes characterized by mitochondrial vacuolization, myofilament lysis, and glycogen accumulation were detected obvious at 3 h post pacing. Down-regulated mRNA expression of Ca(2+) channel beta1 and alpha1 subunits was observed 6 h post pacing, Kv4.3 mRNA down-regulation occurred 24 h post pacing, auxiliary subunit alpha2 was not affected by pacing. Protein expression of alpha1c subunit and potassium channel Kv4.3 paralleled their mRNA expression changes.

CONCLUSIONRAP induced ultrastructural changes of the atrium and down-regulated mRNA and protein expressions of L-type calcium channel subunits and potassium channel Kv4.3 occurred thereafter in response to intracellular calcium overload induced by RAP.

Animals ; Calcium Channels, L-Type ; genetics ; metabolism ; Cardiac Pacing, Artificial ; methods ; Female ; Heart Atria ; metabolism ; ultrastructure ; Male ; Patch-Clamp Techniques ; Potassium Channels ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Rabbits

Adult ; Blood Platelets ; chemistry ; CD40 Antigens ; blood ; CD40 Ligand ; blood ; Cholesterol ; blood ; Female ; Humans ; Hypercholesterolemia ; blood ; drug therapy ; Male ; Middle Aged ; Simvastatin ; therapeutic use

OBJECTIVETo investigate radiation protection and possible mechanisms of low intensity microwave on gamma-ray exposed mice.

METHODS96 healthy Kunming mice were randomly divided into the following four groups: normal control, microwave (120 microW/cm(2), 900 MHz), gamma-ray irradiation (5 Gy), combined exposure of microwave and gamma-ray (120 microW/cm(2) + 5 Gy). The microwave group and combined group were exposed to 120 microW/cm(2) microwave firstly, 1 h/d, for 14 days. Then the ionization and combined group were exposed to 5 Gy (60)Co gamma-ray irradiation on the 15th day. Animals were sacrificed on the third, 6th, 9th and 12th day after irradiation. The sternum and spleen paraffin section were produced, and the histological changes were observed. Apoptosis rate of mice splenic cells in each group was examined by flow cytometry, and serum concentration of antioxidant and lipid peroxide was detected at the same time.

RESULTSBone marrow was obviously injured either by radiation or microwave exposure, characterized by undergoing four-phase lesions, namely apoptosis-necrosis, void, regeneration and recovery phase. Compared with the gamma-ray group, the pathological changes in combined group were slighter and the recovery was quicker. The pathological injuries of spleen were similar to that of bone marrow. Injuries in the combined group were slighter than gamma-ray group. It showed that apoptosis rate of splenic cells in combined group was significantly lower on the 6th and 9th day after gamma-ray radiation (23.02% +/- 15.18%, 25.37% +/- 11.62% respectively) from FCM results. Assays of oxidative damages suggested that serum superoxide dismutase (SOD) level in combined group increased while lipid peroxide level decreased significantly (P < 0.05).

CONCLUSIONLow intensity microwave may exert protection effects on injuries induced by ionizing radiation. The underlying mechanisms might be related with suppression on the hematopoietic cells apoptosis induced by gamma-ray radiation, inhibition of oxidative damages, and thus enhanced reconstruction of the hematopoietic system.

Animals ; Apoptosis ; radiation effects ; Dose-Response Relationship, Radiation ; Gamma Rays ; adverse effects ; Male ; Mice ; Microwaves ; Radiation Protection

Objective: To investigate the relationship between serum HGF levels and clinical severity of essential hypertension (EH). Methods: The serum HGF concentrations of 44 patients with EH were measur ed by ELISA. Results: The serum HGF levels in patients with EH w ere higher than that in control. Furthermore, the serum HGF levels of EH patient s with coronary atherosclerosis (CAS) were significantly higher than those of EH patients without CAS ［(920.8±250.0） pg/ml vs （747.9±132.1） pg/ml, P <0.01］ or control ［（643.8±98.2） pg/ml, P<0.01)］.The changes of HGF l evel were correlated with the clinical courses (r=0.63, P<0.01) and stag es (r=0.69, P<0.01) of hypertension. Conclusion: HGF may be considered as a new index for the severity of hypertension and an useful bio chemical parameter for estimating the development of atherosclerosis.

Objective　To explore the injurious effect of the serum drawn from patient under cardiopulmonary bypass (CPB) on pulmonary surfacant-associated protein A(SP-A) and the preventive effect of pentoxifylline (PTX) on the protein. Methods　The cultured rat alveolar epithelial cell type Ⅱ(AT-Ⅱ) were incubated with CPB serum to observe the change of the cell morphology, and the expressions of SP-A and SP-A mRNA. Results　Traumatic changes and the decrease of SP-A and SP-A mRNA expressions were found in AT-Ⅱ cells cultured with CBP serum. PTX exerted protective effect on the cells. Conclusion　The serum after CPB can directly injure rat AT-Ⅱ cells in vitro, and inhibit the SP-A expression at transcription and translation levels, which probably is an important reason for the quantitative and qualitative abnormality of pulmonary surfactant after operation. PTX may alleviate the inhibitory effect of CPB serum on SP-A.

Biotinylated chitosan nanoparticles (Bio-CS-NP) were prepared for the active delivery to cancer cells and its characterization was investigated in this study. The preparation process included two steps. First, biotinylated chitosan ( Bio-CS ) was obtained through a reaction between sulfosuccinimidobiotin and chitosan (CS). Second, Bio-CS-NP were prepared by the precipitation of Bio-CS with sodium chloride solution. With a biotin reagent box, the conjugation densities of biotin on the surface of Bio-CS-NP were determined. The morphology and diameter of the nanoparticles were assayed by transmission electron microscope (TEM) and laser light scattering particle analyzer, respectively. The uptake of nanoparticles by human hepotacarcinoma HepG2 cells, for example, Bio-CS-NP and chitosan nanoparticles (CS-NP) without any modification, was quantitatively examined. The results indicated that the conjugation densities of biotin on the surface of Bio-CS-NP were 2.2 biotin CS. Bio-CS-NP were spherical, smooth on the surface. The average diameter was 296.8 nm. The polydispersion index was 0.155. The uptake of Bio-CS-NP by HepG2 cells was much higher than that of CS-NP (P < 0.05). It demonstrated that Bio-CS-NP can be applied as a new vehicle to actively deliver anticancer drugs to tumor cells. The method for the determination of biotin was simple and practical.
Biotin ; analogs & derivatives ; chemistry ; metabolism ; Biotinylation ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Chitosan ; chemistry ; metabolism ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Nanoparticles ; Particle Size ; Succinimides ; chemistry ; metabolism

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