RESUMO
Objective: To analyze the expression microarray data in the public databases of intracranial aneurysms (IA) using bioinformatics, and to provide important information for the study of disease mechanisms. Methods: Gene co-expression network was constructed by weighted gene co-expression network analysis (WGCNA) based on the dataset (GSE75436) and pivot genes were identified. Using the online tool DAVID (Database for Annotation, Visualization, and Integrated Discovery) to perform GO function enrichment and KEGG path analysis on modules highly related to IA. Results: Three IA-related modules were screened out, and 14 pivot genes (COL3A1, SPARC, CDH11, COL5A1, HOPX, CLEC11A, GALNT10, ADAMTS2, CEMIP, KIAA1755, COL11A1, ZIC2, CDKN2A, and LINC00460) in the brown module were identified; the analysis of GO showed that the brown module was mainly enriched in extracellular matrix organization, extracellular matrix organization, cell adhesion and other biological processes; the analysis of KEGG indicated that the brown module involved in ECM-receptor interaction, Focal adhesion, protein digestion and absorption, PI3K-Akt signaling pathway. Conclusion: Based on WGCNA, we identified modular and pivotal genes that are critical to the development of IA, and they may become potential biomarkers and/or therapeutic targets.
Assuntos
Aneurisma Intracraniano , Biologia Computacional , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Fosfatidilinositol 3-QuinasesRESUMO
This study aimed to investigate the effect of hydroxy safflower yellow A (HSYA) on myocardial apoptosis after acute myocardial infarction (AMI) in rats. We randomly divided 170 male Wistar rats into 6 groups (N = 23): normal control, sham, control, SY (90 mg/kg), HSYA high-dose (HSYA-H, 40 mg/kg), and HSYA low-dose groups (HSYA-L, 20 mg/kg). Myocardial ischemic injury was induced by ligating the anterior descending coronary artery, and the degree of myocardial ischemia was evaluated using electrocardiography and nitroblue tetrazolium staining. Bax and Bcl-2 expressions in the ischemic myocardium were determined using immunohistochemical analysis. Peroxisome proliferator-activated receptor-γ (PPAR-γ) expression in the myocardium of rats with AMI was determined using reverse transcription-polymerase chain reaction. Compared to rats in the control group, those in the HYSA-H, HSYA-L, and SY groups showed a decrease in the elevated ST segments and an increase in the infarct size. The rats in the drug-treated groups showed a significantly lower percentage of Bax-positive cells and a significantly higher percentage of Bcl-2-positive cells than those in the control group (P < 0.05). Moreover, mRNA expression of PPAR-γ in the ischemic myocardium of rats in the SY, HSYA-L, and HSYA-H groups was significantly lower than that in the control group (P < 0.05). Thus, HSYA and SY can attenuate myocardial ischemia in rats, possibly by increasing the level of Bcl-2/Bax, and PPAR-γ may be not a necessary link in this process.
Assuntos
Apoptose/efeitos dos fármacos , Chalcona/análogos & derivados , Infarto do Miocárdio/metabolismo , Miocárdio/metabolismo , Animais , Chalcona/farmacologia , Eletrocardiografia/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Coração/efeitos dos fármacos , Coração/fisiopatologia , Imuno-Histoquímica , Masculino , Infarto do Miocárdio/genética , Infarto do Miocárdio/prevenção & controle , Isquemia Miocárdica/genética , Isquemia Miocárdica/metabolismo , Isquemia Miocárdica/prevenção & controle , Miocárdio/patologia , Receptores Ativados por Proliferador de Peroxissomo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Distribuição Aleatória , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína X Associada a bcl-2/metabolismoRESUMO
Miscanthus x giganteus is often considered as a suitable plant species for phytomanagement of heavy metal polluted sites. Nevertheless, its physiological behavior in response to the level of metal toxicity throughout the growing season remains poorly documented. Miscanthus x giganteus was cultivated on three sites in Belgium (BSJ: non-polluted control; CAR: slightly contaminated; VM strongly polluted by Cd, Pb, Cu, Zn, Ni and As). The presence of Miscanthus improved soil biological parameters assessed by measurement of enzyme activity and basal soil respiration on the three considered sites, although to a lower level on VM site. Heavy metal accumulation in the shoot was already recorded in spring. It displayed a contrasting distribution in the summer leaves since heavy metals and As metalloid accumulated mainly in the older leaves of CAR plants while showing a uniform distribution among leaves of different ages in VM plants. Comparatively to plants growing on BSJ, net photosynthesis decreased in plants growing on CAR and VM sites. The recorded decrease was mainly related to stomatal factors in CAR plants (decrease in stomatal conductance and in Ci) but to non-stomatal factors such as decrease in carboxylation efficiency and non-photochemical quenching in VM plants. Stomata remained open in VM plants which presented lower instantaneous and intrinsic water use efficiencies than CAR and BSJ plants. High proportions of heavy metals accumulated in CAR plants were bound to the cell wall fraction while the soluble and organelle-rich fractions were proportionally higher in VM plants, leading to a decrease in cell viability and cell membrane damages. It is concluded that not only the intensity but also the nature of physiological responses in Miscanthus x giganteus may drastically differ depending on the pollution level.
RESUMO
Fusarium crown rot (FCR) is a serious cereal disease in semi-arid regions worldwide. In assisting the effort of breeding cultivars with enhanced resistance, we identified several barley genotypes with high levels of FCR resistance. One of these genotypes, AWCS079 which is a barley landrace originating from Japan, was investigated by developing and assessing three populations of recombinant inbred lines. Two QTL, one located on the long arm of chromosome 1H (designated as Qcrs.cpi-1H) and the other on 3HL (designated as Qcrs.cpi-3H), were found to be responsible for the FCR resistance of this genotype. Qcrs.cpi-1H is novel as no other FCR loci have been reported on this chromosome arm. Qcrs.cpi-3H co-located with a reduced height (Rht) locus and the effectiveness of the former was significantly affected by the latter. The total phenotypic variance explained by these two QTL was over 60 %. Significant effects were detected for each of the QTL in each of the three populations assessed. The existence of these loci with major effects should not only facilitate breeding and exploitation of FCR-resistant barley cultivars but also their further characterization based on fine mapping and map-based gene cloning.
Assuntos
Resistência à Doença/genética , Fusarium/fisiologia , Hordeum/genética , Hordeum/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Locos de Características Quantitativas/genética , Mapeamento Cromossômico , Cromossomos de Plantas/genética , Resistência à Doença/imunologia , Hordeum/anatomia & histologia , Hordeum/imunologia , Escore Lod , Doenças das Plantas/microbiologia , Reprodutibilidade dos TestesRESUMO
Traits related to spike morphology (SM), including grain density (GD), spike length (SL) and awn length (AL), are of central importance in cereal improvement. A recent study based on a two-row landrace of barley, TX9425, detected QTL controlling all of the three traits in a similar region on the long arm of chromosome 3H. To further characterize this chromosomal region, 12 pairs of near isogenic lines (NILs) for GD were generated from two populations between TX9425 and two different commercial cultivars. A population consisting of 1,028 lines segregating primarily for the target region was also developed using materials generated during the production of these NILs. Results from the analysis of the NILs and the NIL-derived population showed that these three traits were likely controlled by a single-locus which was mapped to a 2.84 cM interval between two SSR markers, GBM1495 and HVM33. Across the 12 pairs of NILs, the presence of the 3HL locus increased GD by 53.4 %, reduced SL and AL by 38.8 % and 62.7 %, respectively. In the NIL-derived population, the presence of the 3HL locus increased GD by 64.6 %, reduced SL and AL by 33.7 % and 62.6 %, respectively. An interesting question arising from this research is why some loci such as the one reported here affect several SM-related traits while others appear to affect one of these traits only. The NILs and the NIL-derived population generated in this study will help answer such questions by providing the germplasm to enable cloning and comparative analysis of the genes responsible for these SM-related traits.
Assuntos
Cromossomos de Plantas/genética , Hordeum/anatomia & histologia , Hordeum/genética , Endogamia , Locos de Características Quantitativas/genética , Sementes/anatomia & histologia , Sementes/genética , Mapeamento Cromossômico , Segregação de Cromossomos/genética , Ligação Genética , Genética Populacional , GenótipoRESUMO
Crown rot (CR) is one of the most destructive diseases of barley and wheat. Fusarium species causing CR survive in crop residue and a growing acceptance of stubble retention practices has exacerbated disease severity and yield loss. Growing resistant cultivars has long been recognised as the most effective way to reduce CR damage but these are not available in barley. In a routine screening of germplasm, a barley landrace from China gave the best CR resistance among the genotypes tested. Using a doubled haploid population derived from this landrace crossed to Franklin, we demonstrate that the CR resistance of TX9425 was conditioned by a major QTL. The QTL, designated as Qcrs.cpi-3H, was mapped near the centromere on the long arm of chromosome 3H. Its effect is highly significant, accounting for up to 63.3% of the phenotypic variation with a LOD value of 14.8. The location of Qcrs.cpi-3H was coincident with a major QTL conferring plant height (PH) and the effect of PH on CR reaction was also highly significant. When the effect of PH was accounted for by covariance analysis, the Qcrs.cpi-3H QTL remained highly significant, accounting for over 40% of the phenotypic variation. The existence of such a major QTL implies that breeding barley cultivars with enhanced CR resistance should be feasible.
Assuntos
Hordeum/anatomia & histologia , Hordeum/genética , Hordeum/imunologia , Imunidade Inata/genética , Doenças das Plantas/microbiologia , Locos de Características Quantitativas/genética , Animais , Produtos Agrícolas/anatomia & histologia , Produtos Agrícolas/genética , Produtos Agrícolas/imunologia , Produtos Agrícolas/microbiologia , Fusarium/patogenicidade , Predisposição Genética para Doença , Hordeum/microbiologia , Humanos , Doenças das Plantas/imunologia , Distribuição AleatóriaRESUMO
The"Association of Han medicine of State of Manchuria"was a puppet TCM academic society founded by the Puppet Manchukuo government, with well-organization system and widespread scope. During its period, though some efforts were made to promoting the TCM academic progress, improving the quality of TCM doctors, developing TCM clinic, education, academic research and administration, its essence was still a tool for the puppet government to controlling, transforming and utilizing TCM.
RESUMO
Low molecular weight polylactic acid (PLA) microparticles containing prostaglandin E2 were prepared. An average particle size of 30 micron was obtained by grinding at low temperature. These particles were further treated by heating to modify the shape and the release pattern. Microscopic studies showed that the modified particles had a smoother surface than the non-modified particles. The drug was also incorporated into PLA microcapsules using the solvent evaporation process, but the incorporation efficiency was lower. We studied the release profiles of modified particles prepared using different molecular weight PLA. The release rate depended on the molecular weight with lower molecular weights having a greater release rate. In addition, the release studies showed different matrix forms made from the same molecular weight PLA had different release patterns. For example, the microcapsules released the drug very slowly whereas the modified particles exhibited a moderate release rate. It was also noted that the matrix release model could describe the release patterns of microcapsules and modified particles very well. However, the release patterns of non-modified microparticles did not follow this model.
Assuntos
Dinoprostona/farmacocinética , Lactatos , Ácido Láctico , Polímeros , Cápsulas , Dinoprostona/administração & dosagem , Peso Molecular , Tamanho da Partícula , PoliésteresRESUMO
Prostaglandin E2 was successfully incorporated into low molecular weight polylactic acid microcapsules. The size distribution of the microcapsules ranges from 20 to 50 micron. The microcapsules released drug continuously up to three days in vitro. The galactosamine induced fulminant hepatic failure rats model was used. 36 hours after the injection of galactosamine, those rats in grade II coma were chosen and pairs were matched for comparable degree of coma. Then each of the pair was randomly selected as control or treated. Each rat in the treated group received an intraperitoneal injection of PGE2 microcapsules containing 0.55 mg of PGE2. The control received blank PLA microcapsules only. The survival rate of the treated group was 40% compared to 10% in the control group. There was a significant (P less than 0.05) increase in the survival rate in the treated group.
Assuntos
Galactosamina , Encefalopatia Hepática/tratamento farmacológico , Ácido Láctico , Prostaglandinas E/uso terapêutico , Animais , Cápsulas , Preparações de Ação Retardada , Dinoprostona , Encefalopatia Hepática/induzido quimicamente , Lactatos , Masculino , Poliésteres , Polímeros , Prostaglandinas E/administração & dosagem , Ratos , Ratos EndogâmicosRESUMO
We are reporting here that low-mol wt B-cell growth factor (LMW-BCGF) and recombinant interleukin-2 (rIL-2) are together able to induce CD3+ cytotoxic T lymphocytes (CTL) with lymphokine-activated killer cell (LAK) activity from the bone marrow (BM) cells of children with acute lymphoblastic leukemia (ALL). Ficoll-Hypaque (FH)-separated BM cells were obtained from patients with active disease (at diagnosis N = 13, in relapse N = 15) and in complete remission (CR; N = 12). CD3+ cells were removed by Leu-4 antibody and immunobeads. Cells were cultured (10(5) cells/mL) in semisolid media with rIL-2 (100 mu/mL), LMW-BCGF (0.1 mu/mL), and the combination of rIL-2 plus LMW-BCGF, respectively, for seven to ten days. Pooled colonies were harvested for phenotyping. LMW-BCGF plus rIL-2 induced large numbers of CD3+ colonies from CD3- precursors. rIL-2 alone did not induce colony formation. In addition, cells were cultured in liquid media with LMW-BCGF, rIL-2, and the combination of LMW-BCGF plus rIL-2, respectively, for seven to 21 days. They were harvested for phenotyping, and cytotoxicity assays were performed v K562, Raji, and autologous leukemic cells. LMW-BCGF plus rIL-2 induced significant expansion of CD3+ cells from CD3- precursors, and these cells were activated to kill autologous leukemic cells in addition to Raji and K562 cell lines. LMW-BCGF or rIL-2 alone did not induce significant expansion or activation of cytotoxic CD3- cells. Our hypothesis is that LMW-BCGF plus rIL-2 stimulates the proliferation and activation of CD3- precursors from the BM cells of children with acute leukemia to become CD3+ cells that have LAK activity. This finding may have therapeutic implications.
Assuntos
Medula Óssea/imunologia , Citotoxicidade Imunológica/efeitos dos fármacos , Interleucina-2/farmacologia , Interleucinas/farmacologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/imunologia , Linfócitos T Citotóxicos/imunologia , Antígenos de Superfície/análise , Medula Óssea/patologia , Células Cultivadas , Criança , Combinação de Medicamentos , Humanos , Interleucina-4 , Células Matadoras Naturais/imunologia , Peso Molecular , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Proteínas Recombinantes/farmacologia , Ensaio Tumoral de Célula-TroncoRESUMO
The X-gene product of human hepatitis B virus is a transacting transcriptional factor which activates a variety of heterologous viral and host promoters/enhancers. We have found that the X-gene product can significantly transactivate the regulatory sequences located at the 5'-upstream of the c-jun oncogene when a reporter plasmid containing the sequences was co-transfected to HepG2 cells with an X-gene expression plasmid. The results of mutational analysis indicate that the X-gene activation requires the AP-1 sequence of the c-jun gene. Furthermore, we also found that the X-gene is capable of activating the 5'-upstream sequence of the alpha-fetoprotein gene. There are at least two elements that respond to the X-gene transactivation. One is located in the sequences between -5,100 and -2,900, and the other is at the C/EBP site. Therefore, the X-gene activates the c-jun and alpha-fetoprotein genes through different host factors, namely AP-1 and C/EBP, respectively. The results of c-jun activation by the X-gene strongly support the previous hypothesis that the X-gene may play a critical role in the development of hepatocellular carcinoma.
Assuntos
Genes Virais , Antígenos da Hepatite B/genética , Vírus da Hepatite B/genética , Proteínas Proto-Oncogênicas c-jun/genética , Transativadores/genética , alfa-Fetoproteínas/genética , Sequência de Bases , Proteínas Estimuladoras de Ligação a CCAAT , DNA/metabolismo , Proteínas de Ligação a DNA/metabolismo , Humanos , Dados de Sequência Molecular , Proteínas Nucleares/metabolismo , Plasmídeos , Proteínas Proto-Oncogênicas c-jun/metabolismo , Sequências Reguladoras de Ácido Nucleico , Ativação Transcricional , Células Tumorais Cultivadas , Proteínas Virais Reguladoras e AcessóriasRESUMO
MHC-I binding peptides and beta 2 microglobulin (beta 2-m) can upregulate the MHC-I heavy chain expression on certain peptide transporter mutant cells. We have further studied this with normal cells and non-mutant cell lines. No MHC-I upregulation was seen with normal, resting or activated T cells. On mouse cell lines P815 and B16, both peptides and human beta 2-m gave an additive upregulation response. With the human small cell lung carcinoma H82, an optimal HLA.A2 binding peptide (GILGFVFTL) gave an upregulation response, whereas beta 2-m alone or in combination with this peptide had no effect. However, beta 2-m potentiated the response of H82 cells to a slightly longer peptide. Using mutant RMA-S cells, it was found that both Brefeldin A (BFA) and chloroquine, but not leupeptin, inhibited MHC-I upregulation response to both peptide and beta 2-m. In contrast to chloroquine, BFA also gave a reduction of background membrane MHC-I expression, presumably due to a block in Golgi transport. Human beta 2-m, which binds to RMA-S cells, and which is known to internalize into endosomes, did not reappear on the cell surface. When Db on RMA-S cells was upregulated by human beta 2-m, the sensitivity of these cells to Db restricted CTL cells increased. Even if beta 2-m did not upregulate the overall MHC-I expression on normal cells, it may still quantitatively increase the expression of optimally presented peptides and endosomal recycling many be important in this process.
Assuntos
Antígenos de Superfície/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Microglobulina beta-2/imunologia , Sequência de Aminoácidos , Animais , Antibacterianos/farmacologia , Brefeldina A , Cloroquina/farmacologia , Ciclopentanos/farmacologia , Citometria de Fluxo , Humanos , Camundongos , Dados de Sequência Molecular , Peptídeos/imunologia , Linfócitos T Citotóxicos/imunologia , Células Tumorais Cultivadas , Regulação para Cima/efeitos dos fármacosRESUMO
We recently reported that low molecular weight B-cell growth factor (LMW-BCGF) plus recombinant interleukin-2 (rIL-2) synergistically induced lymphokine-activated killer (LAK) activity from the bone marrow (BM) cells of children with acute lymphoblastic leukemia (ALL). The kinetics of cell growth, antigenic phenotype, and lytic activity of the generated effector cells were further analyzed in this study. BM cells from ALL patients with active disease and in complete remission (CR) were cultured with a combination of LMW-BCGF and rIL-2. Monoclonal antibodies (anti-CD3 and anti-Leu 19) and immunomagnetic beads were used to separate LAK cells into three subsets: CD3+/Leu 19-, CD3+/Leu 19+, and CD3-/Leu 19+. Cytotoxicity assays with different subsets were performed versus K562, Raji, and autologous leukemic cells, using a 3-hour 51Cr release test. There was a significant cell expansion of 54-fold (mean value) for CD3+ cells and 15-fold for Leu 19+ cells in culture with LMW-BCGF plus rIL-2 for 7 to 14 days, whereas no cell expansion was observed in culture with rIL-2 alone. Although NK activity (K562) was generated from leukemic BM cells in culture with rIL-2 alone, it is only about one third of that generated in culture with rIL-2 plus LMW-BCGF. Analysis of lytic activity of cells generated in the latter cultures demonstrated that CD3-/Leu 19+ cells expressed highest lytic activity against NK-sensitive K562 cells as well as against NK-resistant Raji cells. CD3+/Leu 19+ cells showed median cytotoxicity, and CD3+Leu 19- cells mediated only minimal cytotoxic activity. Also, lytic activity of CD3-/Leu 19+ cells against autologous leukemic blasts was noted in patients with active disease. Our results demonstrate that LAK activity generated from BM cells by LMW-BCGF and r-IL2 is mediated mainly by two types of Leu 19+ cells: CD3-/Leu 19+ NK cells and CD3-/Leu 19+ T cells. Although CD3+ T cells (both Leu 19+ and Leu 19-) mediated less antitumor cytotoxicity than CD3-/Leu 19+ cells, the former cells were the major expanding cell population in culture with LMW-BCGF and rIL-2. The new culture system may be effective in generation of cells with LAK activity for therapeutic use.
Assuntos
Células Matadoras Ativadas por Linfocina/imunologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/imunologia , Adolescente , Antígenos CD/análise , Medula Óssea/imunologia , Medula Óssea/patologia , Citotoxicidade Imunológica , Antígenos HLA-DR/análise , Humanos , Imunidade Celular , Técnicas In Vitro , Interleucina-2/farmacologia , Interleucina-4/farmacologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologiaRESUMO
Recently, low-molecular-weight B-cell growth factor (LMW-BCGF) has been reported to stimulate growth of leukemic cells from B-cell precursor-acute lymphoblastic leukemia (BCP-ALL). We further investigated the effects of LMW-BCGF on proliferation of leukemic clonogenic (progenitor) and nonclonogenic (progeny) cells from children with BCP-ALL (28 patients) and B-cell ALL (two patients). Patients were either at diagnosis (n = 18) or in relapse (n = 12). Response of leukemic progenitor cells was determined by culturing cells (10(5) cells/mL) in methylcellulose with 0.1 U/mL LMW-BCGF. Colonies (greater than 20 cells) were counted at day 7. The response of the leukemic progeny population was determined by DNA synthesis studies using tritiated-thymidine and by DNA quantitation with propidiumiodide for determination of cell-cycle status. LMW-BCGF supported growth of leukemic progenitor cells from 20 of 28 (71%) BCP-ALL and two of two B-cell ALL patients. Colony numbers ranged from 7 to 2,400 (mean 145, median 45). A dose-response effect in colony growth was noted, with an apparent plateau at approximately 2.0 U/mL LMW-BCGF. Colony cells were primarily of leukemic phenotype (CD19+/CD10+/-). LMW-BCGF also induced significant increases in leukemic progeny cell proliferation as measured by both thymidine incorporation (stimulation indexes of 1.6 to 34) and by cell-cycle assay (percentage S+ G2/M stimulation indexes of 1.6 to 6). LMW-BCGF was more effective in stimulating leukemic proliferation than three recombinant interleukins (rIL-2, rIL-3, rIL-4), although rIL-3 was able to support colony growth in 4 of 11 patients. These results indicate that LMW-BCGF and, to a lesser degree rIL-3, are able to stimulate proliferation of BCP-ALL progenitor and progeny cells, whereas rIL-2 and rIL-4 do not support progenitor cell proliferation and have only marginal effects on leukemic progeny cell proliferation.