CpG array analysis of histone H3 lysine 4 trimethylation in patients with IgA nephropathy / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the aberrance of histone H3 lysine 4 trimethylation (H3K4me3) in patients with IgA nephropathy (IgAN).</p><p><b>METHODS</b>In 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.</p><p><b>RESULTS</b>We 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.</p><p><b>CONCLUSION</b>IgAN patients have significant alterations in H3K4me3, and the genes with aberrant H3K4me3 may provide insights into the pathogenesis of IgAN.</p>

The study of the differential gene expression profiles related to toxic effects in rats exposed to silica / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the differential gene expression profiles related to toxic effects in rats exposed to silica.</p><p><b>METHODS</b>Wistar rats exposed to SiO2 (50 mg/ml) and 1 ml normal saline by intratracheal injection served as the exposure and control groups, on the 14th day after exposure all rats were executed and the rat lung tissues were obtained. The differential gene expression profiles in the lung tissues of rats exposed to silica were detected using confocal fiber beads gene chip technique, and the differential expression profiling data were analyzed using the database for annotation, visualization and integrated discovery (DAVID) bioinformation analysis tool.</p><p><b>RESULTS</b>The results of present study indicated that 1567 genes with differential expression were identified in 22107 genes of rat lung tissues in exposure group, including 765 up-regulated genes and 802 down-regulated genes as compared to control group. In the 461 genes related to toxic effects, 285 genes were up-regulated and 176 genes were down-regulated in exposure group. The trends of up-regulation of HMOX1 and SOD2 genes in RT-PCR assay were similar to those in gene chip technique.</p><p><b>CONCLUSION</b>A large number of genes related to toxic effects in the rats with silica-induced pulmonary fibrosis appeared up-regulation or down-regulation. There may be a complex gene regulation network in the pulmonary fibrosis induced by SiO2, and the toxicological mechanism is an important part in the development of pulmonary fibrosis.</p>

Cytogenetic and molecular characterization of partial trisomy 4q and partial monosomy 10q in a patient / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To ascertain the karyotype of a girl with moderate mental retardation and growth retardation, perform correlation analysis between chromosomal variation and phenotype, and investigate the application and superiority of array-based comparative genomic hybridization (array-CGH) in clinical cytogenetic diagnosis.</p><p><b>METHODS</b>G-banded chromosome analysis, array-CGH, fluorescence in situ hybridization (FISH) and real-time quantitative PCR (RQ-PCR) were used to ascertain the karyotype of the patient and her relatives.</p><p><b>RESULTS</b>G-banding analysis of the patient showed a derivative chromosome 10 with an extra fragment on its long arm terminal, both her father and grandmother had an apparently balanced translocation t(4;10)(q25;q26). Array-CGH revealed that the breakpoint on chromosome 4 was located at 4q26. In addition, a microdeletion of about 0.54 Mb del(10)(q26.3) was identified from the patient. FISH and RQ-PCR confirmed that the del(10)(q26.3) was also present in both her father and grandmother.</p><p><b>CONCLUSION</b>No recognizable phenotype was associated with del(10)(q26.3). The abnormal phenotypes presented in the patient may be ascribed to the 4q26-q35.2 triplication. Further more, compared with conventional cytogenetic analysis, array-CGH is of high resolution and high accuracy.</p>

Effect of hepatocyte growth factor gene transfection on biological features of lymphoma cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the biological effect of hepatocyte growth factor (HGF) on HGF gene-transfected Raji cells.</p><p><b>METHODS</b>Total RNA was extracted from human hepatic tissue, HGF gene cDNA was amplified by RT-PCR, and then cloned into vector pVITRO2-mcs to construct recombinant eukaryotic expression vector pVITRO2-mcs-HGF. The recombinant vector was transfected to Raji cells, and the stably transfected cells were selected by homomycin B in serial passages, and confirmed by real-time fluorescent quantitative PCR, ELISA, immunocytohistochemistry. The biological features of transfected Raji cells were evaluated by semisolid culture.</p><p><b>RESULTS</b>RT-PCR results showed that Raji cells were transfected successfully with recombinant eukaryotic expression vector pVITRO2-mcs-HGF. HGF mRNA and protein were expressed successfully in Raji cells. Expression of HGF gene enhanced proliferation, metastasis and invasion of Raji cells.</p><p><b>CONCLUSION</b>HGF gene has been cloned and recombined to construct recombinant eukaryotic expression vector pVITRO2-mcs-HGF successfully. Transfected HGF may change the biological features of Raji cells.</p>

Predictive value of human fatty acid binding protein for myocardial ischemia and injury in perioperative period of cardiac surgery / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the value of human fatty acid binding protein (h-FABP) in predicting myocardial ischemia and injury in the perioperative period of cardiac surgery, we observed the dynamic changes of h-FABP in perioperative period of patients underwent coronary artery bypass grafting and ventricular septal defects repairing surgery, and evaluated the relationship of h-FABP and ischemia modified albumin (IMA), CK-MB, cTnI.</p><p><b>METHODS</b>Patients underwent coronary artery bypass grafting (n=30) and ventricular septal defect repairing (n=30) surgery between February 2008 and December 2008 were included in this study. Venous blood sample was obtained at preoperative, aortic clamping, aortic unclamping of 10 min, 2 h, 6 h, 12 h, 24 h for the measurements of h-FABP, IMA, cTnI and CK-MB.</p><p><b>RESULTS</b>h-FABP and IMA changed in the same way at various examined time points, h-FABP changes also paralleled cTnI and CK-MB changes, h-FABP peaked early during myocardial ischemia and injury and returned to baseline level at 2 h post myocardial ischemia and injury. Linear correlation analysis showed that the peak value of h-FABP was positively correlated with IMA, CK-MB and cTnI in both CABG group (r = 0.948, 0.964 and 0.961, P < 0.05) and in the VSD group (r = 0.986, 0.978 and 0.957).</p><p><b>CONCLUSIONS</b>h-FABP is an early diagnostic parameter reflecting perioperative myocardial ischemia and injury in cardiac surgery. Quantitative h-FABP monitoring could predict the severity of myocardial ischemia and injury early during cardiac surgery.</p>

The study on differential gene expression profiling in pulmonary tissue of rats exposed to silica early / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the differential gene expression profiling of rats exposed to silica using the normal rats as control.</p><p><b>METHODS</b>Animal models were established using intratracheal injection of the lung and 22 107 genes were screened in the differential expression profiling of silicosis by using oligonucleotide bead array. Differential expression profiling data were analyzed by using DAVID bioinformation software.</p><p><b>RESULTS</b>Totally 1567 differentially expressed genes were identified in lungs of silica exposed rats including 765 up-regulated genes and 802 down-regulated genes as compared to the normal controls. Among 406 annotated genes in KEGG pathways, 204 genes and 11 pathways were up-regulated as well as 202 genes and 3 pathways were down-regulated in silica exposed rats.</p><p><b>CONCLUSION</b>All 1567 genes are involved in the formation of silicosis. The differential gene expression profile of silicosis describes the general changes in the gene expressions in silicosis at transcriptional level. Further analysis of the identified genes might help reveal the molecular mechanism of pulmonary fibrosis induced by silica.</p>

Changes of Clara cell protein and surfactant protein-D in serum of patients with silicosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore changes of Clara cell protein (CC16) and surfactant protein-D (SP-D) in the serum of patients with silicosis.</p><p><b>METHOD</b>The 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.</p><p><b>RESULTS</b>The 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%.</p><p><b>CONCLUSION</b>The serum CC16 of long-term silica-exposed workers is decreased, and SP-D is increased gradually.</p>

Utilizing 2-DE and MALDI-TOF MS/MS to screen differentially expressed serum proteins of silicosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To establish 2-dimensional gel electrophoresis (2-DE) images and seek differentially expressed serum proteins for understanding the pathogenesis of silicosis.</p><p><b>METHODS</b>2-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).</p><p><b>RESULTS</b>Complement 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.</p><p><b>CONCLUSION</b>Complement 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.</p>

Determination of serum argininosuccinate lyase in diagnosing liver diseases / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the value of the determination of the levels of serum argininosuccinate lyase (ASL) in diagnosing various liver diseases.</p><p><b>METHODS</b>Two 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.</p><p><b>RESULTS</b>Receiver 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).</p><p><b>CONCLUSION</b>ASL 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.</p>

Detection and clinical analysis of CD_(62p)and CD_(63)in cancer patients / 肿瘤研究与临床

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

Secretory-expression of Antimicrobial Peptide Bactenecin7 Gene in Lactococcus lactis and Analysis the Bioactivity of Its Expression Products / 中国生物工程杂志

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.

VEGF antisense oligonucleotide inhibits the expression of vascular endothelial growth factor in human leukemic cell lines / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effects of vascular endothelial growth factor (VEGF) antisense phosphorothioated oligodeoxynucleotide (AS-ODN) on the expression of VEGF in human leukemic cell lines (HL-60 and K562 cells).</p><p><b>METHODS</b>The levels of VEGF mRNA and protein in leukemic cells incubated with VEGF AS-ODN were measured by RT-PCR, immunohistochemistry assay and ELISA. MTT test was used to examine the influence of the culture supernatant (CS) of VEGF AS-ODN treated leukemic cells on the proliferation of human umbilical vein endothelial cells (ECV304).</p><p><b>RESULTS</b>After leukemic cells were treated with different concentrations (2.5 approximately 15.0 micro mol/L) of VEGF AS-ODN for 24 h, VEGF mRNA level in the cells decreased remarkably in a concentration dependent manner, no change was found in the VEGF missense ODN treated cells (MS-ODN). When the leukemic cells were treated with 5 micro mol/L VEGF AS-ODN for 24 h, VEGF protein level decreased greatly both in the cells and in the CS; and the proliferation stimulating effect of the treated CS on the ECV304 cells reduced. Meanwhile, there was no obvious change in VEGF protein and its effect in the VEGF MS-ODN treated group.</p><p><b>CONCLUSION</b>VEGF AS-ODN could inhibit VEGF expression in human leukemic cell lines in vitro.</p>

Targeted blockage of STAT5 by a decoy oligodeoxynucleotide inhibits the growth and proliferation of K562 cells / 中华血液学杂志

<p><b>OBJECTIVES</b>To 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.</p><p><b>METHODS</b>STAT5 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.</p><p><b>RESULTS</b>Confocal 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.</p><p><b>CONCLUSIONS</b>STAT5 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.</p>

Study on the transcriptional activation of MTS1 gene beta promoter / 中华血液学杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Seven 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.</p><p><b>RESULTS</b>Seven 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.</p><p><b>CONCLUSION</b>MTS1 gene beta promoter can be activated in Jurkat cell line.</p>