RESUMO
The present study aimed to explore the genetic changes involved in the liver hepatocellular carcinoma (HCC) development. The RNA-Seq data of 212 HCC tissue samples and 50 normal tissue samples were downloaded using TCGA-Assembler. A total of 4 subgroups were obtained, and 4167, 6279, 5379, and 2548 DEGs were screened in group 1, group 2, group 3, and group 4, respectively. Enrichment analysis found that cell cycle, metabolism, and translation related terms were the most significantly changed functions and pathways. There were 454 genes (1114 pairs), 803 genes (722 pairs), and 788 genes (724 pairs), separately interacted in the condition specific PPI network of group 1, 2, 3, and 4, with MMP2, ATNXN1, F2, and HDAC1 as the hub genes. What's more, using these genes, total 7, 20, 198, and 1 subtype related miRNAs; 35, 50, 47, and 17 subtype related TFs; 1, 1, 0, and 2 subtype related drugs were screened in group 1, 2, 3, and 4, respectively. The integrated biological analysis on RNA-Seq data provided substantial of bio-molecular related to the HCC development. miR-147b, SP1, and Riboflavin were the subtype-related regulator/drug for HCC. The study about the big data of HCC RNA-Seq data reveals the intrinsic gene expression pattern of the tumor, which provides a novel perspective to understand the heterogeneity of pathogenesis in HCC tumorigenesis.
Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , RNA Neoplásico/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Análise de Sequência de RNARESUMO
OBJECTIVE: To investigate the long-term therapeutic effect of autologous hematopoietic stem cell transplantation in patients with End-stage Liver Disease (ESLD). PATIENTS AND METHODS: Forty-eight ESLD patients underwent autologous CD34+ stem cell transplantation were retrospectively reviewed. Changes in clinical and biochemical data, complications, and quality of life were monitored at 3, 6, 12, 36, and 60 months following the stem cell transplantation. Liver biopsies were obtained for histopathological analysis using Ishak system. RESULTS: Marked improvement in clinical and biochemical data was observed during the long-term follow-up. Serum albumin was significantly increased (p<0.001), while total serum bilirubin, prothrombin time (PT), and international normalized ratio (INR) were all significantly decreased (p<0.001). Ishak inflammation and fibrosis scores were significantly decreased with the increased time (p<0.001). The number of patients with ascites, model of end-stage liver disease (MELD) score, Child-Pugh class, and indocyanine green (ICG) score were all markedly reduced with increased time. Meanwhile, the quality of life score of the patients was significantly increased (p<0.001). Six patients died during the 5-years follow-up, and complications occurred in 17 patients. The incidence of complications was significantly associated with mortality of the patients (p<0.05). CONCLUSIONS: The study provided the evidence that autologous CD34+ stem cell transplantation could offer a long-term therapeutic benefit to patients with ESLD. The complications occurred during the process was significantly associated with survival of the patients. Future studies on a large cohort of patients are needed to confirm the long-term effect of stem cell therapy on ESLD.
Assuntos
Doença Hepática Terminal/terapia , Transplante de Células-Tronco Hematopoéticas , Doença Hepática Terminal/fisiopatologia , Doença Hepática Terminal/psicologia , Seguimentos , Humanos , Testes de Função Hepática , Transplante de Fígado , Qualidade de Vida , Estudos RetrospectivosRESUMO
Androgen plays critical roles in vertebrate reproductive systems via androgen receptors (ARs). In the present study, the full-length spotted scat (Scatophagus argus) androgen receptor (sAR) cDNA sequence was cloned from testis. The sAR cDNA measured 2448 bp in length with an open-reading frame of 2289 bp, encoding 763 amino acids. Amino acid alignment analyses showed that the sARs exhibited highly evolutionary conserved functional domains. Phylogenetically, the sARs clustered within the ARß common vertebrate group. Real-time polymerase chain reaction (RT-PCR) revealed that sAR expression varied in level and distribution throughout the tissues of both females and males. sAR expression was detected during testicular development by quantitative RT-PCR. The results showed that the highest transcription of sARs was observed in the mid-testicular stage, and remained at a high expression level until the late-testicular stage. In addition, the effects of 17α-methyltestosterone (MT) and estrogen (E2) on the expression of sARs in ovaries were determined using quantitative RT-PCR. sAR expression increased at 12 and 24 h post-MT treatment and decreased with E2 treatment. The present study provides preliminary evidence indicating gonadal plasticity of spotted scat under exogenous steroidal hormone treatments. It also provides a theoretical basis for sex reversal and production of artificial pseudo-males for female monosex breeding.
Assuntos
Proteínas de Peixes/genética , Perciformes/genética , Receptores Androgênicos/genética , Animais , Clonagem Molecular , Estrogênios/farmacologia , Feminino , Proteínas de Peixes/química , Proteínas de Peixes/metabolismo , Gônadas/efeitos dos fármacos , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Masculino , Metiltestosterona/farmacologia , Fases de Leitura Aberta , Perciformes/metabolismo , Domínios Proteicos , Receptores Androgênicos/química , Receptores Androgênicos/metabolismoRESUMO
The gonad-inhibiting hormone (GIH) belongs to a neuropeptide family synthesized and released in an X-organ sinus gland complex of crustacean eyestalks. GIH inhibits crustacean ovarian maturation by suppressing vitellogenin (Vtg) synthesis, whereas estrogen is responsible for the stimulation of vitellogenesis (not established). In this study, the effects of 17ß-estradiol (E2, 10(-6) M), estrogen receptor antagonist tamoxifen (TAM, 10(-6), 10(-7), and 10(-8) M), and the environmental estrogen nonylphenol (NP, 1 µg/L and 100 µg/L) on LvGIH expression in the eyestalks of shrimp were determined by quantitative real-time PCR. Results showed that LvGIH expression decreased significantly during the L. vannamei ovarian maturation cycle. E2 and NP significantly reduced LvGIH transcripts in vivo, but TAM neutralized the inhibitory action of E2 in a dose-dependent manner (P < 0.05). In addition, the LvGIH expression levels decreased significantly in a time-dependent manner (P < 0.05) when ovary fragments were cultured in vitro with E2. The results of this study suggested that estrogen regulates GIH expression in L. vannamei eyestalks. E2 promoted ovarian development not only by directly upregulating vitellogenesis in the hepatopancreas, but it was also capable of downregulating LvGIH expression, which indirectly resulted in the stimulation of L. vannamei vitellogenesis.
Assuntos
Proteínas de Transporte/biossíntese , Estradiol/farmacologia , Hormônios de Invertebrado/biossíntese , Penaeidae/efeitos dos fármacos , Fenóis/farmacologia , Animais , Proteínas de Transporte/genética , Antagonistas de Estrogênios/farmacologia , Estrogênios/metabolismo , Feminino , Expressão Gênica/efeitos dos fármacos , Hormônios de Invertebrado/genética , Ovário/efeitos dos fármacos , Ovário/metabolismo , Penaeidae/genética , Penaeidae/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Tamoxifeno/farmacologia , Vitelogênese/efeitos dos fármacos , Vitelogeninas/metabolismoRESUMO
In this study, the complete foxl2 complementary (c)DNA sequence was isolated by simple modular-architecture research tool (SMART)er rapid amplification of cDNA ends (RACE). Two year-old female spotted scat, Scatophagus argus, were reared at different temperatures (23, 26 and 29° C) for 6 weeks, or fed with different concentrations of dietary fish oil (0, 2 or 6%) for 8 weeks. Ovarian development, serum oestradiol-17ß (E2 ) levels, as well as ovarian foxl2 expression were measured. At the end of experiment, ovarian foxl2 messenger (m)RNA expression in fish reared at 23 and 26° C was significantly higher than that in fish reared at 29° C, and that in 2 and 6% fish oil groups was also significantly higher than that in control group (P < 0·05). Serum E2 levels exhibited the same trend with foxl2 mRNA expression in temperature treatment groups and fish oil fed groups. There was a significant positive correlation between stage of oocytes and foxl2 expressions. Results showed that from 23 to 29° C, the optimal temperature for ovarian development in S. argus was 23-26° C, and 6% fish oil supplementation could effectively promote ovarian development. Optimal temperature and fish oil supplement might increase ovarian foxl2 mRNA expressions to promote ovarian development in S. argus.
Assuntos
Suplementos Nutricionais , Óleos de Peixe , Fatores de Transcrição Forkhead/genética , Ovário/crescimento & desenvolvimento , Perciformes/crescimento & desenvolvimento , Temperatura , Animais , Estradiol/sangue , Feminino , Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento , Oócitos/crescimento & desenvolvimento , Perciformes/genética , RNA Mensageiro/genética , Diferenciação SexualRESUMO
OBJECTIVES: Lycoris is aurea agglutinin (LAA) has attracted rising attention due to its remarkable bioactivities. Here, we aimed at investigating its anti-tumor activities. MATERIAL AND METHODS: In vitro methods including MTT, cellular morphology observation, FCM and immunoblotting were performed. In vivo methods like detection of tumor volume, body weight and survival ratio, as well as TUNEL staining were performed. RESULTS AND CONCLUSION: LAA triggers G2 /M phase cell cycle arrest via up-regulating p21expression as well as down-regulating cdk-1cyclinA singling pathway, and induces apoptotic cell death through inhibiting PI3K-Akt survival pathway in human lung adenocarcinoma A549 cells. While LAA has no significant cytotoxic effect toward normal human embryonic lung fibroblast HELF cells, and moreover, LAA could amplify the antineoplastic effects of cisplatin toward A549 cells. Lastly LAA also bears anti-cancer and apoptosis-inducing effects in vivo, and it could decrease the volume and weight of subcutaneous tumor mass obviously as well as expand lifespan of mice. These findings may provide a new perspective for elucidating the complicated molecular mechanisms of LAA-induced cancer cell growth-inhibition and death, providing a new opportunity of LAA as a potential candidate anti-neoplastic drug for future cancer therapeutics.
Assuntos
Adenocarcinoma/metabolismo , Apoptose/efeitos dos fármacos , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Neoplasias Pulmonares/metabolismo , Lycoris/metabolismo , Adenocarcinoma de Pulmão , Aglutininas/farmacologia , Antineoplásicos/farmacologia , Proteína Quinase CDC2/biossíntese , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cisplatino/farmacologia , Ciclina A/biossíntese , Inibidor de Quinase Dependente de Ciclina p21/biossíntese , Humanos , Modelos Moleculares , Simulação de Acoplamento Molecular , Inibidores de Fosfoinositídeo-3 Quinase , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidoresRESUMO
Currently lifelong immunosuppression is required for organ transplant recipients. Anti-CD45RB monoclonal antibody (mAb) prolongs graft survival by mechanisms that are not yet clear. Therefore, we investigated the role of T and dendritic cells (DC) in islet allografts treated with anti-CD45RB mAb after transplantation of 200 allogeneic islets (BALB/c mouse) under the kidney capsules of diabetic C57BL/6 mice treated with intraperitoneal injections of 100 µg of anti-CD45RB mAb on days 0, 1, 3, 5, and 7. We observed a tilt of the ratios of Th1/Th2 and Tc1/Tc2 to Th2 and Tc2. The numbers of naïve and memory T cells were down-regulated in peripheral blood after transplantation. In addition, the maturation, endocytosis, and interleukin-12 secreted by DC derived from bone marrow cells was suppressed in recipient mice. Therefore, anti-CD45RB mAb alleviated, rejection by suppressive effects on T-lymphocyte subsets and DC.
Assuntos
Anticorpos Monoclonais/imunologia , Células Dendríticas/imunologia , Ilhotas Pancreáticas/imunologia , Antígenos Comuns de Leucócito/imunologia , Linfócitos T/imunologia , Animais , Diabetes Mellitus Experimental/imunologia , Citometria de Fluxo , Memória Imunológica , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Estreptozocina , Transplante HomólogoRESUMO
Changes in soil microbial community structure and diversity may reflect environmental impact. We examined 16S rRNA gene fingerprints of bacterial communities in six agroecosystems by PCR amplification and denaturing gradient gel electrophoresis (PCR-DGGE) separation. These soils were treated with manure for over a century or different fertilizers for over 70 years. Bacterial community structure and diversity were affected by soil management practices, as evidenced by changes in the PCR-DGGE banding patterns. Bacterial community structure in the manure-treated soil was more closely related to the structure in the untreated soil than that in soils treated with inorganic fertilizers. Lime treatment had little effect on bacterial community structure. Soils treated with P and N-P had bacterial community structures more closely related to each other than to those of soils given other treatments. Among the soils tested, a significantly higher number of bacterial ribotypes and a more even distribution of the bacterial community existed in the manure-treated soil. Of the 99 clones obtained from the soil treated with manure for over a century, two (both Pseudomonas spp.) exhibited 100% similarity to sequences in the GenBank database. Two of the clones were possible chimeras. Based on similarity matching, the remaining 97 clones formed six major clusters. Fifty-six out of 97 were assigned taxonomic units which grouped into five major taxa: alpha-, beta-, and gamma-Proteobacteria (36 clones), Acidobacteria (16 clones), Bacteroidetes (2 clones), Nitrospirae (1 clone), and Firmicutes (1 clone). Forty-one clones remained unclassified. Results from this study suggested that bacterial community structure was closely related to agroecosystem management practices conducted for over 70 years.
Assuntos
Bactérias/genética , Bactérias/isolamento & purificação , Ecossistema , Esterco/microbiologia , Microbiologia do Solo , Agricultura , Animais , Bactérias/classificação , Sequência de Bases , Impressões Digitais de DNA , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Genes Bacterianos , Variação Genética , Dados de Sequência Molecular , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Fatores de TempoRESUMO
Glucokinase and phosphoenolpyruvate carboxykinase are key enzymes of glucose metabolism in the rat liver. The former is considered to be instrumental in regulating glucose hepatic release/uptake according to the glycaemia level, and cytosolic phosphoenolpyruvate carboxykinase is a major flux-generating enzyme for gluconeogenesis. The level of expression of both enzymes and the regulation of their mRNAs in the human liver cell were investigated. Surgical biopsies of liver from patients undergoing partial hepatectomies and parenchymal hepatocytes derived from the biopsies were used to assay glucokinase, hexokinase and phosphoenolpyruvate carboxykinase activities. Hepatocytes were placed in culture and the actions of insulin, glucagon and cAMP on glucokinase and phosphoenolpyruvate carboxykinase mRNAs were studied. The main results are: (a) glucokinase accounts for 95% of the glucose phosphorylation activity of human hepatocytes, although this fact is masked in assays of total liver tissue; (b) glucokinase activity is set at a lower level in human hepatocytes than in rat hepatocytes, and vice-versa for the gluconeogenic enzyme phosphoenolpyruvate carboxykinase; and (c) as previously shown in rat liver, glucokinase and phosphoenolpyruvate carboxykinase mRNAs are regulated in a reciprocal fashion in human hepatocytes, insulin inducing the first enzyme and repressing the latter, whereas glucagon has opposite effects. These data have interesting implications with respect to metabolic regulation and intracellular hormone signaling in the human liver.
Assuntos
Regulação Enzimológica da Expressão Gênica , Glucoquinase/biossíntese , Fígado/enzimologia , Fosfoenolpiruvato Carboxiquinase (GTP)/biossíntese , Adulto , Animais , Sequência de Bases , Biópsia , Células Cultivadas , Criança , Técnicas de Cultura/métodos , AMP Cíclico/farmacologia , Citosol/enzimologia , Primers do DNA , Indução Enzimática/efeitos dos fármacos , Feminino , Glucagon/farmacologia , Humanos , Isoenzimas/biossíntese , Cinética , Fígado/patologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , RNA Mensageiro/biossíntese , RatosRESUMO
A complementary DNA for a glucagon-like peptide-1 receptor was isolated from a human pancreatic islet cDNA library. The isolated clone encoded a protein with 90% identity to the rat receptor. In stably transfected fibroblasts, the receptor bound [125I]GLP-1 with high affinity (Kd = 0.5 nM) and was coupled to adenylate cyclase as detected by a GLP-1-dependent increase in cAMP production (EC50 = 93 pM). Two peptides from the venom of the lizard Heloderma suspectum, exendin-4 and exendin-(9-39), displayed similar ligand binding affinities to the human GLP-1 receptor. Whereas exendin-4 acted as an agonist of the receptor, inducing cAMP formation, exendin-(9-39) was an antagonist of the receptor, inhibiting GLP-1-induced cAMP production. Because GLP-1 has been proposed as a potential agent for treatment of NIDDM, our present data will contribute to the characterization of the receptor binding site and the development of new agonists of this receptor.
Assuntos
Expressão Gênica/genética , Fragmentos de Peptídeos/farmacologia , Peptídeos/farmacologia , Receptores de Superfície Celular , Receptores de Glucagon , Peçonhas , Sequência de Aminoácidos , Aminoácidos/análise , Sequência de Bases , Clonagem Molecular , AMP Cíclico/análise , AMP Cíclico/metabolismo , DNA/análise , DNA/genética , Diabetes Mellitus Tipo 2/tratamento farmacológico , Exenatida , Receptor do Peptídeo Semelhante ao Glucagon 1 , Humanos , Ilhotas Pancreáticas/química , Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/ultraestrutura , Ligantes , Dados de Sequência Molecular , Fragmentos de Peptídeos/análise , Peptídeos/análise , Peptídeos/uso terapêutico , Receptores de Superfície Celular/antagonistas & inibidores , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/fisiologiaRESUMO
An automated clean up and concentration method by column switching is described for the assay of gliquidone, a hypoglycemic agent, in plasma. The system uses Merck Lichroprep RP2 (25-40 microns) as short precolumn packing material for on--line clean up and concentration. A Shimpack CLC--ODS (5 microns) is used as analytical column. Water is used as the pretreatment mobile phase and a mixture of methanol--isopropanol--0.2 mol/L ammonium acetate solution (61:5:34) is used as the analytical mobile phase. The plasma is diluted with a solution of 0.2 mol/L acetic acid and injected directly on the precolumn. The low speed of 0.2 ml/min of pretreatment mobile phase during transfer of plasma sample on the precolumn can get recovery of 80.7%. The detection is at 310 nm and the detection limit is 30 ng/ml plasma. The procedure is simple and automatic.
Assuntos
Hipoglicemiantes/sangue , Compostos de Sulfonilureia/sangue , Cromatografia Líquida de Alta Pressão/métodosRESUMO
Using small dosage of human growth hormone to immunize rabbit or guinea pig, it is able to induce anti-hGH antibody formation with high titre and high affinity that could be applied to hGH RIA. In the present study five rabbits and three guinea pigs were immunized with 125-200 micrograms and 250-285 micrograms per animal of hGH respectively, followed by boosters of 10-20 or 160-250 micrograms of hGH at 2-4 week intervals for 6 or 3 months. Blood was drown 1-2 weeks before each booster for determination of antibody formation. Antibody titre and affinity were successively observed and specificity of antibody was determined for the final bleeding. It was shown that titres of immune sera from guinea pigs were much higher than those of rabbit immune sera, but vice versa for antibody affinity. This might be due to larger immunogen dose used for guinea pigs than for rabbits. Fourteen different peptide hormones were tested in reference to cross-immunoreactivity to anti-hGH antibody. It could be demonstrated that the major cross-reactive hormones are hFSH and hLH, and hTSH also reacts to rabbit anti-hGH immune sera at a lesser degree. These cross reactivities are obviously owing to the molecular homogeneities between hGH and these hormones especially of their alpha-subunits.