Animals ; Humans ; Microchip Analytical Procedures ; Neoplasms ; pathology ; Neoplasms, Experimental ; pathology ; Pathology ; Periodicals as Topic ; RNA Interference

Objective:To prepare and characterize specific and discrepant mouse hybridoma antibodies on membrane of HL60 and HL60/ADR cell lines.Methods:BALB/c mice were immunized by subtractive immunization induced Cp(Cyclophosphamide).McAbs were prepared by hybridoma technique,screened and detected by FACS and LSCM.Results:51 candidates and discrepant antibodies were found,and one of them (5F6) was purified and identified.Conclusion:Combination of SI with discrepant screening method should facilitate the preparing and identifying discrepant McAbs for identifying antibodies that can distinguish the differences in proteins expressed in HL60 and HL60/ADR,which is a significative and potential method in the research and target therapy associated drug-resistance.

OBJECTIVETo evaluate the preventive and therapeutic effect of thymosin alpha(1) on lung infections in critical patients with tracheotomy.

METHODSForty-two patients were randomly divided into treatment group and control group to receive daily subcutaneous thymosin injection at 11.6 mg and saline of 2 ml for 7 days, respectively.

RESULTSCompared with the control group, the infection rate, white blood cell count, C-reactive protein, tumor necrosis factor-alpha and interleukiu-6 were significantly lower in the treatment group.

CONCLUSIONThymosin alpha(1) can be effective for prevention and treatment of lung infections in critical patients with tracheotomy and may improve the patients' immunity and prognosis.

Adjuvants, Immunologic ; therapeutic use ; Adolescent ; Adult ; Aged ; Brain Injuries ; drug therapy ; surgery ; Cerebral Infarction ; drug therapy ; Critical Illness ; Female ; Humans ; Intensive Care Units ; Male ; Middle Aged ; Pneumonia ; prevention & control ; Thymosin ; analogs & derivatives ; therapeutic use ; Tracheotomy ; adverse effects

OBJECTIVETo review the biological behaviour of endothelial progenitor cells and their role in vascular diseases. Data sources The data used in this review were mainly from Medline and PubMed for relevant English language articles published from 1985 to March 2007. The search term was "endothelial progenitor cells". Study selection Articles about the biological behaviour of endothelial progenitor cells and their roles in the pathogenesis of vascular diseases such as atherogenesis were used.

RESULTSProgenitor cells in bone marrow, peripheral blood and adventitia can differentiate into mature endothelial cells (ECs). The progenitor cells, which express certain surface markers including AC133, CD34 and KDR, enable restoration of the microcirculation and ECs when injury or ischaemia occurs. Endothelial progenitor cells used in experimental models and clinical trials for ischaemic syndromes could restore endothelial integrity and inhibit neointima development. Moreover, their number and functional properties are influenced by certain cytokines and atherosclerotic risk factors. Impairment of the progenitor cells might limit the regenerative capacity, even lead to the development of atherosclerosis or other vascular diseases.

CONCLUSIONSEndothelial progenitor cells have a particular role in prevention and treatment of certain cardiovascular diseases. However, many challenges remain in understanding differentiation of endothelial progenitor cells, their mobilization and revascularization.

Animals ; Atherosclerosis ; physiopathology ; therapy ; Cardiovascular Diseases ; therapy ; Cell Differentiation ; Cell Separation ; methods ; Endothelial Cells ; cytology ; physiology ; Humans ; Neovascularization, Physiologic ; Stem Cell Transplantation ; Stem Cells ; physiology

BACKGROUNDStudies have shown that oxidized low density lipoprotein (ox-LDL) promotes the pathogenesis and development of atherosclerosis (AS), and that the proliferation, migration and phenotype alteration of vascular smooth muscle cells (vSMCs) into foam cells are critical changes in AS. It is proposed that ox-LDL might play a novel role in the pathologic process of vSMCs. The present study was performed ex vivo to investigate the effects of ox-LDL on the growth of cultured human vSMCs.

METHODSUsing NaBr density gradient centrifugation, LDL from human plasma was isolated and purified. ox-LDL was produced from LDL after being incubated with CuSO4. ox-LDL was then added to the culture medium at different concentrations (25 microg/ml, 50 microg/ml, 75 microg/ml, 100 microg/ml, 125 microg/ml, and 150 microg/ml) for 7 days. The influence of ox-LDL on vSMC growth was observed from several aspects as growth curve, mitosis index, lipid staining, and in situ determination of apoptosis. The digital results were analyzed with SPSS 10.0.

RESULTSThe ox-LDL produced ex vivo had a good purity and optimal oxidative degree, which was similar to the intrinsic ox-LDL in atherosclerotic plaque. ox-LDL at a concentration of 25 microg/ml demonstrated the strongest proliferation. At the concentration of 125 microg/ml, ox-LDL suppressed the growth of vSMCs. At concentrations of 25 microg/ml and 50 microg/ml, ox-LDL presented powerful mitotic trigger. When the concentration of ox-LDL increased, the mitotic index of vSMCs decreased gradually. ox-LDL induced more foam cells from vSMCs with rich intracellular lipid accumulation at concentrations of 25 microg/ml and 50 microg/ml. ox-LDL at higher concentrations induced more apoptotic vSMCs.

CONCLUSIONSox-LDL at lower concentrations may trigger proliferation and phenotype alteration into foam cells of vSMCs, and at higher concentrations it may induce apoptosis in vSMCs. ox-LDL plays an important role in the pathogenesis and development of atherosclerosis by its effect on vSMCs proliferation, phenotype alteration and apoptosis.

Apoptosis ; drug effects ; Atherosclerosis ; etiology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Humans ; Lipoproteins, LDL ; toxicity ; Mitotic Index ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; cytology ; drug effects

Haploinsuffieiency of the runt-related transcription factor 2 (Runx2) gene is widely known to be responsible for cleidocranial dysplasia (CCD).To date,more than 190 mutations in Runx2 gene have been reported to be related to CCD.In this study,a novel mutation of Runx2 gene was observed in a female with CCD.Genomic DNA was extracted from peripheral venous blood of the proband and eleven members of her family.Genetic testing on these twelve people identified a novel missense mutation (c.895T＞C,Y299H) in exon 5 of the RUNX2 gene in the proband.This mutation results in an amino acid change at codon 895 (P.Tyr 299 His.) from a tryptophan codon (TAT) to a histidine codon (CAT).Our finding may further extend the known mutation spectrum of the RUNX2 gene,and facilitate prenatal genetic diagnosis of CCD in the future.

OBJECTIVETo detect whether the activation of nuclear factor-kappa B (NF-kappaB) in endothelial cells induced by mm-LDL can promote platelet-derived growth factor-B (PDGF-B) expression in vitro, and whether it is also present in hypercholesterolemic rats in vivo, influence of age on NF-kappaB and PDGF-B signal transduction pathway.

METHODSEstablished hypercholesterolemic rat model by feeding with a high-cholesterol ration. The activation of NF-kappaB in aortic endothelial cells was identified by immunohistochemical staining, the expression of PDGF-B mRNA and PDGF-B protein were examined using in situ hybridization and immunohistochemistry respectively.

RESULTSIn comparison with the control rats, a positive immunostaining of NF-kappaB in nuclei of aortic endothelial cells of the experimental rats was detected after a high cholesterol ration for 6 weeks. The number of endothelial cells expressing PDGF-B mRNA increased and the intensity was dependent upon the duration of high-cholesterol intake. NF-kappaB translocation (0.461 +/- 0.075 vs. 0.350 +/- 0.094, P < 0.05) and PDGF-B expression in 10-month old Wistar rats were more remarkable than that of 2-month old rats after having cholesterol for 16 weeks. Immunohistochemical staining for PDGF-B gave a similar result (0.230 +/- 0.040 vs. 0.185 +/- 0.037, P < 0.001).

CONCLUSIONSHypercholesterolemia is capable of activating nuclear translocation of NF-kappaB and promoting expression of PDGF-B in rat aortic endothelial cells in vivo, this coincided with the results obtained in ox-LDL or mm-LDL experiments on endothelial cells in vitro. This phenomenon is much more evident in 10-month old rats which indicates that age might have a close relationship with NF-kappaB - PDGF-B signal transduction pathway.

Active Transport, Cell Nucleus ; Age Factors ; Animals ; Aorta ; metabolism ; Arteriosclerosis ; etiology ; Endothelial Cells ; metabolism ; Hypercholesterolemia ; metabolism ; Immunohistochemistry ; Male ; NF-kappa B ; metabolism ; Proto-Oncogene Proteins c-sis ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

OBJECTIVETo study the effect of high-density lipoprotein (HDL) on the proliferation of endothelial progenitor cells (EPC) isolated from human umbilical cord blood; to further explore its effect on prevention and development of atherogenesis.

METHODSEPC isolated by density gradient centrifugation were cultured in a M200 medium. Immunofluorescence staining for CD133, CD34, KDR and Factor VIII were adopted respectively as the specific markers for identification. The effect of HDL on EPC proliferation was estimated on the 7th day of cell cultivation using MTT assay, confocal microscopy and fluorescence activated cell sorting.

RESULTSHDL, when incubated with EPC, was able to promote remarkably the proliferation rate of EPC, dose- and time-dependent. HDL participated in the transcriptional regulation of cell cycle by affecting the regulatory proteins such as cyclin D1.

CONCLUSIONSA subtype of progenitor cells was isolated from human cord blood with a potential of differentiating into mature endothelial cells (known as endothelial progenitor cells). HDL plays an important role on EPC fluorescence activated cell sorting differentiation and proliferation. Further studies are required to identify the signal pathway and the molecular mechanism of HDL effect on EPC proliferation.

AC133 Antigen ; Adult ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cell Separation ; Cells, Cultured ; Cyclin D1 ; metabolism ; Dose-Response Relationship, Drug ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Factor VIII ; metabolism ; Fetal Blood ; cytology ; Flow Cytometry ; Fluorescent Antibody Technique ; Glycoproteins ; metabolism ; Humans ; Lipoproteins, HDL ; blood ; pharmacology ; Microscopy, Confocal ; Peptides ; metabolism ; Stem Cells ; cytology ; drug effects ; metabolism

OBJECTIVETo evaluate the efficacy of hepatitis B viruse (HBV) vaccination and its influencing factors among children in rural area of Jiangsu province.

METHODSTwenty-five hundred and twenty-two children born after 1998 in rural area were selected as the study population using multistage cluster sampling method. HBsAg and anti-HBs were detected by enzyme linked immunoassay (ELISA) and radio-immunoassay (RIA), respectively. Anti-HBs negative children were boosted using different hepatitis B vaccines and the efficacy was compared. Factors causing HBV infection in HBsAg positive children were also investigated.

RESULTSHBsAg positive rates in 1-7 year olds were 0.28%-1.28%, and the anti-HBs positive rates decreased from 76.7% to 45.5%. The HBsAg positive rate in children not timely vaccinated was significantly higher than those with HBV vaccine injection within 24 hours after birth (1.4% vs. 0.5%, P = 0.031). More than 90% of the anti-HBs negative children had protective level of anti-HBs after boosted with HBV vaccine.

CONCLUSIONHBsAg positive rate in children born after 1998 in rural area of Jiangsu province decreased significantly, with an average of 0.8%. The reason for HBsAg carriage in children might be attributed to mother-to-infant transmission or not timely HBV vaccination.

Adult ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hepatitis B ; epidemiology ; immunology ; prevention & control ; transmission ; Hepatitis B Surface Antigens ; blood ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Humans ; Infant ; Infectious Disease Transmission, Vertical ; Pregnancy ; Rural Population

OBJECTIVETo evaluate the effects of rapamycin (RPM)-loaded poly (lactic-co- glycolic) acid (PLGA) nanoparticles (NPs) on the proliferation, distribution of cell cycle, and expression of p27 protein in human umbilical arterial vascular smooth muscle cell (HUASMC) in vitro.

METHODSThe primarily culture model of HUASMC was successfully established by explant-attached method in vitro. The cells were administrated with different doses of RPM, and RPM-PLGA NPs were observed as treat groups compared with PLGA NPs and M231-SMGs medium cultured group. The effect of RPM-PLGA NPs on proliferation of HUASMC was assessed using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) colorimetry method. The influences of RPM-PLGA NPs on the cell cycle and cellular growth kinetics of HUASMCs were tested by flow cytometry. The effect of RPM-PLGA NPs on the expression of p27 protein of HUASMCs was assessed through an immunohistochemical method.

RESULTSCompared with the control group, the proliferation of HUASMCs was inhibited by 50 microg/L and higher concentration of RPM-PLGA NPs in a dose-dependent manner (P < 0.05). The numbers of cells entering cell cycle of S/G2/M phases were significantly lower in RPM-PLGA NPs and RPM treated groups. Histologically, the expression of p27 were up-regulated in 500 microg/L RPM-PLGA NPs and 100 microg/L RPM treated group (all P < 0.01 ) when compared with the control group.

CONCLUSIONSRPM-PLGA NPs has a similar effects as RPM in inhibiting the growth of in vitro cultured HUASMC. It can remarkably suppress the expression of in vitro cultured HUASMC p27 protein, arrest its cell cycle at G1/S phase, and inhibit its proliferation.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Drug Carriers ; Humans ; Lactic Acid ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Nanoparticles ; Polyglycolic Acid ; Sirolimus ; administration & dosage ; pharmacology ; Umbilical Arteries ; cytology