Objective To compare the measured values of activated platelets in different disease sta- tus,and the prevention and treatment effects of aspirin for thromboembolism.Methods Flow cytometry was used to detect CD_(62p)and CD_(63)level in peripheral activated platelets(PAP) of 126 cases of tumor patients and 60 cases of non-tumor patients, and detect the changes after taking aspirin. Results The CD_(62p)and CD_(63) levels of PAP were evidently higher in tumor patients than that of non-tumor patients(P

To construct a secretory-expression vector of antimicrobial peptide Bactenecin 7(Bac7),and identify the secretory-expression product in L.lactis MG1363 and its bioactivity.The splicing primers of regulation elements and Bac7 gene,which designed according to codon usage preferences of L.lactis MG1363,were chemically synthesized,and the overlap-extension PCR method was used to splice the full length of Bac7 gene.Then the Bac7 gene was linked to expression vector pMG36e to construct pMG36e/Bac7 vector,and pMG36e/Bac7 was transformed into L.lactis MG1363 by electrophoration.RT-PCR and Western blot assays were applied to investigate the expression of the Bac7 gene in L.lactis,and bioactivity of Bac7 in culture supernatant of L.lactis was tested with plate-diffusion method.The results showed that the Bac7 gene and its regulation elements was amplified and cloned in the vector pMG36e successfully,The secretory-expressed Bac7 in L.lactis MG1363 harboring pMG36e/Bac7 was identified by Western blot,and it had high bacteriostatic activity against E.coli.These results indicate that the recombinant L.lactis MG1363 could express bioactive Bac7,which lays a foundation for further study of oral administration of a Bac7-secreting L.lactis to treat intestinal bacteria infection.

OBJECTIVETo investigate the effect of MTS1 gene beta promoter transcriptional activation in T-cell acute lymphoblastic leukemia (T-ALL) cell lines and identify the fragment with transcriptional activation.

METHODSSeven pGL3 recombinant plasmids with the same 3'-end transcriptional start site but the different 5'sequences were constructed by gene recombinant technique and transfected into Jurkat cell line which is biallelic deletion of MTS1 gene by transient transfection. Luciferase report gene was detected to observe beta promoter transcriptional activation.

RESULTSSeven pGL3 recombinant plasmids containing different fragments of beta promoter were obtained, all of them showed transcriptional activation in Jurkat cell line. Among them, the 0.38 kb fragment cut by SacII-SacI is fundamental in transcription.

CONCLUSIONMTS1 gene beta promoter can be activated in Jurkat cell line.

Genes, p16 ; Humans ; Jurkat Cells ; Plasmids ; genetics ; Promoter Regions, Genetic ; genetics ; Transcriptional Activation ; Transfection

Carcinoma, Hepatocellular ; pathology ; Cell Transformation, Neoplastic ; drug effects ; Ginsenosides ; pharmacology ; Humans ; Liver Neoplasms ; pathology ; Signal Transduction ; drug effects

OBJECTIVETo investigate the aberrance of histone H3 lysine 4 trimethylation (H3K4me3) in patients with IgA nephropathy (IgAN).

METHODSIn 15 patients with IgAN and 15 healthy volunteers, H3K4me3 variations in peripheral blood mononuclear cells (PBMCs) were analyzed using chromatin immunoprecipitation and microarray analysis (ChIP-chip). ChIP real-time PCR was used to validate the microarray results. Quantitative real-time PCR (qRT-PCR) was carried out to examine the correlations between the mRNA expression profiles and H3K4me3 levels.

RESULTSWe identified 83 genes that displayed significant H3K4me3 differences in IgAN patients compared with healthy subjects. Among them, 39 genes showed increased H3K4me3 and 44 genes had decreased H3K4me3 levels. The results of ChIP real-time PCR were well consistent with the microarray data. Quantitative RT-PCR revealed the correlations between the mRNA expressions and the methylation levels of H3K4me3.

CONCLUSIONIgAN patients have significant alterations in H3K4me3, and the genes with aberrant H3K4me3 may provide insights into the pathogenesis of IgAN.

Case-Control Studies ; CpG Islands ; genetics ; DNA Methylation ; Female ; Glomerulonephritis, IGA ; genetics ; metabolism ; Histones ; genetics ; metabolism ; Humans ; Leukocytes, Mononuclear ; metabolism ; Lysine ; genetics ; metabolism ; Male

OBJECTIVETo establish 2-dimensional gel electrophoresis (2-DE) images and seek differentially expressed serum proteins for understanding the pathogenesis of silicosis.

METHODS2-DE and matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-MS/MS) were used to screen differentially expressed serum proteins among silica-exposed population, suspect of silicosis (0+), phase one (I) group with silicosis and control group(non silica exposure).

RESULTSComplement C4, leucine-rich alpha-2-glycoprotein and alpha-1-antitrypsin were significantly highly expressed in suspect of silicosis (0+) group(P < 0.01), but lowly in other groups. Inversely, serotransferrin was significantly down-regulated only in suspect of silicosis (0+) group(P < 0.01). Plasma glutathione peroxidase, tetranectin, apolipoprotein A-I and transthyretin were equally expressed in the serum of control group and silica-exposed population group, but decreased in the suspect of silicosis (0+) and phase (I) group.

CONCLUSIONComplement C4, leucine-rich alpha-2-glycoprotein, alpha-1-antitrypsin, serotransferrin, plasma glutathione peroxidase, tetranectin, apolipoprotein A-I and transthyretin are differentially expressed in the silica-exposed group and phase (I) group with silicosis, and the result should be validated by other biochemical technologies.

Adult ; Aged ; Aged, 80 and over ; Blood Proteins ; analysis ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Middle Aged ; Proteomics ; methods ; Silicosis ; blood ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo explore changes of Clara cell protein (CC16) and surfactant protein-D (SP-D) in the serum of patients with silicosis.

METHODThe concentrations of CC16 and SP-D were measured in the serum by sandwich enzyme-linked immunosorbent assays. The subjects consisted of 30 healthy volunteers and 90 silica-exposed workers including silica-exposed group, the silicosis of suspects group (0(+)) and the silicosis phase I group, 30 subjects each groups.

RESULTSThe concentrations of CC16 in the serum was significantly decreased in silica-exposed workers compared to controls (P < 0.01); The concentrations of CC16 in the serum were higher in lifelong nonsmokers than the current smokers in control subjects (P < 0.05), but they were no differences between lifelong nonsmokers and current smokers of 90 silica-exposed workers. Compared with control subjects, the levels of SP-D in the serum of silicosis suspects (0(+)) and silicosis phase I groups were significantly elevated (P < 0.01, respectively), which were also higher than silica-exposed group (P < 0.05 and P < 0.01, respectively), Discriminant equations set by CC16 and SP-D were used in diagnosis of silicosis, and the rate of accuracy in healthy volunteers, the silica-exposed group and the silicosis phase I group were 86.7%, 86.7% and 76.7%, respectively, The total rate of correct classification hit 84.2%.

CONCLUSIONThe serum CC16 of long-term silica-exposed workers is decreased, and SP-D is increased gradually.

Adult ; Case-Control Studies ; Epithelial Cells ; metabolism ; Humans ; Male ; Middle Aged ; Pulmonary Surfactant-Associated Protein D ; blood ; Silicosis ; blood ; Uteroglobin ; blood

Cell Line, Tumor ; Gene Silencing ; Hep G2 Cells ; Humans ; RNA, Small Interfering ; STAT3 Transcription Factor ; genetics

OBJECTIVESTo investigate targeted blockage of BCR/ABL oncoprotein mediated cell transformation by STAT5 decoy oligodeoxynucleotide (ODN), its effect on the growth and proliferation inhibition of K562 cells and the related molecular mechanisms.

METHODSSTAT5 decoy ODN, designed and synthesized in vitro, was transfected into K562 cells by cationic lipid. The cell growth curve and colony formation assay were used to reflect the growth and proliferation capacity of K562 cells, RT-PCR to detect the expression of three genes downstream STAT5.

RESULTSConfocal microscopy demonstrated that STAT5 decoy ODN was successfully transfected into K562 cells (95.2% positive cells). STAT5 decoy ODN inhibited the growth of K562 cells (inhibition rate 77.7%) and their colony formation capacity (Decoy ODN treated group 8.3% vs control group 35.7%, P < 0.05) after the treatment with STAT5 decoy ODN, the expressions of c-myc, bcl-X(L), cyclin D1 mRNA were down-regulated by 15.4%, 30.8%, 29.1%, respectively in the K562 cells.

CONCLUSIONSSTAT5 decoy ODN inhibits the growth and proliferation of K562 cells. The mechanisms may be that decoy ODN blocks the transcriptional activation potent of STAT5 and down-regulates the expression of these tumor related genes downstream STAT5.

Cell Proliferation ; Cyclin D1 ; genetics ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Humans ; K562 Cells ; Liposomes ; Microscopy, Confocal ; Oligodeoxyribonucleotides, Antisense ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; STAT5 Transcription Factor ; genetics ; physiology ; Transfection ; bcl-X Protein ; genetics

OBJECTIVETo investigate the value of the determination of the levels of serum argininosuccinate lyase (ASL) in diagnosing various liver diseases.

METHODSTwo hundred and ninety-one patients with various liver diseases, 257 patients with non-liver disease, and 32 healthy controls were recruited for this study and their serum ASL, ALT, AST, GGT, LDH, ALP, and total bilirubin (TBil) levels were determined. Liver biopsies were performed on 31 patients with hepatopathy.

RESULTSReceiver operating characteristic (ROC) curve analysis showed that the sensitivity and specificity of ASL in assessing liver diseases were 100% and 91.1% (at cut-off values of 8 U/L), those of ALT were 97.6% and 24.7% and those of AST were 83.8% and 28.3% (both at cut-off values = 40.0 U/L), respectively. The levels of ASL in various liver disease patients were: in liver cancer - acute hepatitis - liver cirrhosis - chronic hepatitis. A positive correlation (r = 0.417) was observed between serum ASL levels (86.9+/-26.5) and scores of histopathological inflammation grading (9.83+/-3.36).

CONCLUSIONASL is of higher sensitivity and specificity than those of ALT and AST for diagnosing liver diseases. ASL may be used as a useful marker in estimating hepatopathy.

Adolescent ; Adult ; Aged ; Argininosuccinate Lyase ; blood ; Automation ; Female ; Humans ; Liver Diseases ; blood ; diagnosis ; Male ; Middle Aged ; ROC Curve ; Sensitivity and Specificity ; Serum ; chemistry ; Young Adult