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Objective: To explore the clinical manifestations, imaging features, pathological features, diagnosis and treatment of pulmonary mucosal-associated lymphoid tissue(MALT)lymphoma concurrent with lung squamous cell carcinoma, and to improve the understanding of this disease. Methods: Using "Pulmonary mucosa-associated lymphoid tissue, squamous cell carcinoma" as the search term, from January 1, 1983 to August 31, 2020, a total of 3 cases were retrieved in the PubMed database. In the Wanfang database, using "Lung mucosa-associated lymphoid tissue, lung squamous cell carcinoma" as the search term, from January 1, 1990 to August 31, 2020, a total of 1 related document was retrieved. In the CNKI database, "(lung) mucosa-associated lymphoid tissue lymphoma, (lung) squamous cell carcinoma" was used as the search term, and no relevant case reports were retrieved. Results: A 64-year-old man was admitted to the hospital because of chest tightness and shortness of breath for 10 days, cough and fever for one day. Enhanced CT of the chest showed a soft tissue mass shadow in the right lower hilar area, with obstruction of the adjacent bronchus, and local mild enhancement, suggesting of right lower lung cancer. In addition, the CT scan also showed consolidated shadows in the lower lobes of both lungs, scattered nodules, multiple lymphadenopathy in the mediastinum, and a small amount of pleural effusion on the right. Under bronchoscopy, a cauliflower-like neoplasm was seen at the opening of the lower right basal section, about 7 mm×8 mm, and biopsy showed that part of the mucosal structure was destroyed, with disappearance of the squamous epithelial layer, and the nuclei were large and deeply stained, and some were distributed in nests, with poor keratinization and a small amount of necrosis, and fibrous tissue reaction. Immunostaining revealed that the tumor was positive for p40, CK5/6 and EGFR and negative forTTF-1, NapsinA, PD-L1, p53, with about 30% Ki-67 positive cells. A puncture biopsy of the right lower lobe showed that the alveolar cavity was filled with nested lymphoid cells, consisting of small lymphocytes, central cell-like cells and monocyte-like cells, with occasionally large cells. Immunostaining revealed CD20+, CD79a+, scattered CD3+, Bcl2+, SMA vascular+, Bcl6-, CK-, CD10-, CyclinD1-, with about 3% Ki-67 positive cells. The histopathological examinations confirmed the diagnosis of mucosal-associated lymphoid tissue extranodal marginal zone lymphoma(MALT lymphoma),and lung squamous cell carcinoma. Conclusions: Pulmonary mucosa-associated lymphoid tissue lymphoma complicated with lung squamous cell carcinoma is rare and easy to be missed and misdiagnosed. Chest CT imaging shows single or multiple nodules, mass shadows or consolidation, often accompanied by air-bronchial signs in the lesion, bronchiectasis, ground glass density around the lesion, hilar and mediastinal lymphadenopathy. Occasionally, pleural effusion can be seen. Lung biopsy is the gold standard for diagnosis.
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Carcinoma de Células Escamosas/patologia , Neoplasias Pulmonares/patologia , Pulmão/diagnóstico por imagem , Linfadenopatia/diagnóstico por imagem , Linfoma de Zona Marginal Tipo Células B/patologia , Biópsia , Broncoscopia , Carcinoma de Células Escamosas/diagnóstico por imagem , Tosse/etiologia , Dispneia/etiologia , Febre/etiologia , Humanos , Neoplasias Pulmonares/diagnóstico por imagem , Linfoma de Zona Marginal Tipo Células B/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVE: To examine the changes in the immune functions of CD8+ T cells in the spleen of mice following Echinococcus multilocularis infections at various doses and at different time points. METHODS: The E. multilocularis protoscoleces were collected, and E. multilocularis infection was modeled in mice via the hepatic portal vein at doses of 50 (low-dose), 500 (medium-dose) and 2 000 protoscoleces (high-dose), while physiological saline served as controls. Mouse spleen was isolated 2 (earlystage), 12 (middle-stage) and 24 weeks post-infection (late-stage), and spleen lymphocytes were harvested. The phenotype of memory CD8+ T cells and 2B4 expression were quantified in the mouse spleen, and the secretion of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-17A and IL-10 was measured. RESULTS: A central-memory phenotype was predominant in the CD8+ T cells in the spleen of mice at the early stage of high-dose protoscolece infections, and the proportion of central-memory CD8+ T cells was significantly greater in the high-dose group than in the control group (35.50% ± 2.00% vs. 25.90% ± 2.46%, P < 0.01), while a effector- memory phenotype was predominant in the CD8+ T cells in the spleen of mice at the late stage of medium- and high-dose protoscolece infections, and the proportions of effector-memory CD8+ T cells were significantly greater in the medium- (25.70% ± 4.12%) and high-dose group (28.40% ± 4.12%) than in the control group (10.50% ± 6.45%) (P < 0.05). The proportions of the central-memory CD8+ T cells were significantly higher in the high-dose group than at middle and late stages than at the early stage (P < 0.01), and the proportion of effector-memory CD8+ T cells was significantly greater in the high-dose group at the late stage than at early and middle stages (P < 0.05). The secretion of IFN-γ and IL-17A by spleen CD8+ T cells was elevated in the low- and medium-dose groups at the early stage of infection, and high-dose protoscolece infection promoted the secretion of IFN-γ and TNF-α by spleen CD8+ T cells; however, the levels of IFN-γ and TNF-α were significantly lower at the late stage than at the early and middle stages (P < 0.05). In addition, high 2B4 expression was detected in spleen CD8+ T cells in the middle- and high-dose groups at the late stage of infection, and the 2B4 expression was significantly higher in the medium(4.73% ± 1.56%) and high-dose groups (4.94% ± 1.90%) than in the low-dose group (2.49% ± 0.58%) and the control group (2.92% ± 0.60%) (P < 0.05). CONCLUSIONS: E. multilocularis may be killed and eliminated through the host immune responses at the middle and late stages of low- and medium-dose protoscolece infections, while high-dose protoscolece infections may trigger the upregulation of 2B4 expression in mouse spleen CD8+ T cells at the late stage, which leads to immune exhaustion and the resultant chronic infections.
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Linfócitos T CD8-Positivos/imunologia , Equinococose/imunologia , Baço/imunologia , Animais , Citocinas/imunologia , Echinococcus multilocularis , Camundongos , Baço/parasitologiaRESUMO
OBJECTIVE: Liver cancer is the second most common cause of cancer death, causing more than 700,000 deaths every year. It has been demonstrated that Long non-coding RNA (LncRNA) plays an important regulatory role in a series of diseases. However, the regulatory mechanism of LncRNAs in liver cancer has not been fully elucidated. The purpose of this study was to explore the interaction of lncRNA HOTAIRM1 and aberrant histone modification in liver cancer. MATERIALS AND METHODS: qRT-PCR was used to detect the expression levels of RIZ1 and miR-125b in liver cancer cells. Cell proliferation was measured using the CCK8 assay. ChIP-Real-time PCR confirmed the binding site of the promoter of HOTAIRM1 by H3K9me1. The direct target of HOTAIRM1 and miR-125b in liver cancer cells was measured by a luciferase reporter assay. Cell proliferation was detected by Cell Counting Kit-8 (CCK8). Cell invasion was measured by transwell assays and cell migration was detected by wound healing assay. RESULTS: The expression level of RIZ1 and miR-125b was upregulated, and HOTAIRM1 was downregulated in liver cancer cells. Transwell and CCK-8 assay showed that RIZ1 expression is associated with the proliferation, invasion and migration of liver cancer cells, silencing of RIZ1 inhibited cell proliferation, migration, and invasion in HEPG2 and HCC-LM3 cells. RIZ1 interference could significantly inhibit H3K9me1 expression. H3K9me1 protein can bind to HOTAIRM1 promoter directly. Furthermore, the bioinformatics prediction and luciferase assay demonstrated that miR-125b can interact with HOTAIRM1 by direct binding. HOTAIRM1 down-expression promoted HEPG2 cell growth and metastasis, which was further strengthened following the co-transfection of miR-125b. Furthermore, overexpressed HOTAIRM1 inhibited HCC-LM3 cell growth and metastasis and a complete reversal of the results seen when transfected with miR-125b. CONCLUSIONS: For the first time, we found that RIZ1 was upregulated in liver cancer cells and RIZ1-mediated H3K9me1 enrichment on the HOTAIRM1 promoter regulated the growth and metastasis of liver cancer cells by targeting miR-125b, which could further accelerate tumor proliferation, migration and invasion. It may serve as a therapeutic marker for liver cancer treatment.
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Proteínas de Ligação a DNA/genética , Histona-Lisina N-Metiltransferase/genética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , MicroRNAs/genética , Proteínas Nucleares/genética , Fatores de Transcrição/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação para Baixo , Humanos , Regulação para CimaRESUMO
Neutrophil extracellular trap (NET) is a type of bead-like, fibrous and reticular substances that is actively released by activated inflammatory neutrophils during the stage of infections or inflammatory responses. NET, which is composed of chromatin DNA and multiple intracellular protein components, may wrap pathogens to limit their diffusions. Meanwhile, NET may kill pathogens via a wide range of antibacterial proteins, which is considered as the third antibacterial mechanism of neutrophils, in addition to phagocytosis and degranulation. Recent studies have shown the involvement of NET in the immune response against parasitic infections. This review summarizes the advances of NETs in the immune responses against parasitic infections, so as to provide insights into the elucidation of the pathogenesis and development of therapeutics of parasitic diseases.
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Armadilhas Extracelulares , Neutrófilos/imunologia , Doenças Parasitárias/imunologia , DNA , Humanos , ImunidadeRESUMO
OBJECTIVE: The aim of this research was to explore the effect of microRNA-133a (miR-133a) on myocardial fibrosis and cardiac function after myocardial infarction in rats, and to investigate the possible regulatory mechanism. MATERIALS AND METHODS: Myocardial infarction model was successfully established in rats by ligation of the left anterior descending coronary artery. After miR-133a overexpression in rats myocardium, cardiac function was examined by echocardiography. Meanwhile, the degree of myocardial fibrosis was detected by Masson staining. In addition, the expression of α-smooth muscle actin (α-SMA) in cardiomyocytes was detected by immunohistochemistry. Quantitative Real-time polymerase chain reaction (qRT-PCR) was performed to analyze the expression level of miR-133a in the junction of myocardial infarction. The mRNA expressions of transforming growth factor-ß1 (TGF-ß1), connective tissue growth factor (CTGF), collagen type 1 (col 1), collagen type 3 (col 3) and α-SMA were measured by qRT-PCR as well. Furthermore, the protein levels of the above genes were detected by Western blotting. RESULTS: MiR-133a expression in the infarct border zone of myocardial tissue was significantly decreased on the 28th day after myocardial infarction surgery (p<0.05). In addition, up-regulation of miRNA-133a in myocardial tissue of rats with myocardial infarction could remarkably improve cardiac function and reduce collagen volume fraction. Furthermore, the mRNA and protein expression levels of TGF-ß1, CTGF, col1, col3, α-SMA in myocardial tissue were obviously decreased after miRNA-133a up-regulation (p<0.001). CONCLUSIONS: Overexpression of miR-133a down-regulates the mRNA and protein levels of TGF-ß1 and CTGF after myocardial infarction. Moreover, this may eventually reduce myocardial collagen deposition, inhibit myocardial fibrosis and improve cardiac function.
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Cardiomiopatias/metabolismo , Fibrose/metabolismo , MicroRNAs/metabolismo , Infarto do Miocárdio/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Doença Aguda , Animais , Cardiomiopatias/diagnóstico , Cardiomiopatias/cirurgia , Modelos Animais de Doenças , Ecocardiografia , Fibrose/diagnóstico , Fibrose/cirurgia , Masculino , MicroRNAs/genética , Infarto do Miocárdio/diagnóstico , Infarto do Miocárdio/cirurgia , Ratos , Ratos Sprague-Dawley , Transdução de SinaisRESUMO
BACKGROUND: Orexin A, a small-molecule peptide, can regulate female hormones, but limited evidence for its mechanism of activity exists in ovine. AIMS: The objective of this study was to investigate the effect of orexin A on progesterone (P4) secretion in cultured granulosa of sheep follicles. METHODS: Sheep ovarian granulosa were isolated and identified, pre-incubated with luteinizing hormone (LH) (2.5 IU/ml), follicle-stimulating hormone (FSH) (2.5 IU/ml), or oestrogen (1 µg/ml); and cultured in vitro. The pretreated sheep ovarian granulosa were subsequently cultured with different concentrations (1 nM, 10 nM, 58 nM, 100 nM, and 145 nM) of orexin A for varying amounts of time (0 h, 24 h, 48 h, and 72 h). Then, the expression levels of P4, steroidogenic acute regulatory protein (StAR), 3ß-Hydroxysteroid dehydrogenase (3ß-HSD) and cytochrome P450 (CYP11) were determined. RESULTS: The results showed that the sheep ovarian granulosa were correctly identified. The different concentrations of orexin A promoted the secretion of P4 from granulosa in the ovine ovary compared with that in the control. The expression of StAR, 3ß-HSD and P450 (CYP11) gradually increased, and then decreased with increasing concentrations of orexin A, but the expression of P450 (CYP11) decreased with the increase of time. CONCLUSION: These results revealed that orexin A promotes the secretion of P4 by regulating the expression of StAR, 3ß-HSD, and P450 (CYP11). Understanding the mechanism underlying the promotion of P4 by orexin A could open new therapeutic possibilities in the treatment of hormone homeostasis.
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OBJECTIVE: To elucidate the regulatory effect of hypoxic preconditioning bone marrow mesenchymal stem cells (BMSCs)-exosomes on cardiomyocyte apoptosis in acute myocardial infarction (AMI) rats. MATERIALS AND METHODS: BMSCs-derived exosomes were extracted by Exoquick method. Expressions of exosome surface markers were determined by Western blot. The AMI model in rats was established by LAD ligation. Rats were randomly assigned into sham group, AMI group, AMI+H-exo group and AMI+N-exo group. MicroRNA-24 expression in rat myocardium was detected at different time points. Subsequently, hypoxic preconditioning or normoxic preconditioning BMSCs-exosomes were intramyocardially injected into rats. Infarct size was calculated through TTC (triphenyltetrazolium chloride) staining. Cardiomyocyte apoptosis was accessed with Terminal Deoxynucleotidyl Transferase dUTP Nick-end Labeling (TUNEL). Heart function of AMI rats was evaluated by echocardiography. Protein expressions of apoptotic genes in rat myocardium were detected by Western blot. RESULTS: The mRNA level of microRNA-24 was higher in H-exo group than N-exo group. Injection of hypoxic preconditioning BMSCs-exosomes markedly upregulated microRNA-24 level, reduced infarct size and improved cardiac function in AMI rats. Protein expressions of Bax, caspase-3 and cleaved-caspase-3 were downregulated by BMSCs-exosomes treatment. H9c2 cells showed upregulated microRNA-24 level and decreased apoptotic rate after incubation with hypoxic preconditioning BMSCs-exosomes. The above cellular performances were partially reversed by transfection of microRNA-24 inhibitor. CONCLUSIONS: Hypoxic preconditioning BMSCs-exosomes inhibit cardiomyocyte apoptosis in AMI rats by upregulating microRNA-24.
Assuntos
Apoptose/genética , Exossomos , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , Infarto do Miocárdio/terapia , Miocárdio/patologia , Animais , Apoptose/efeitos dos fármacos , Caspase 3/genética , Técnicas de Cultura de Células , Hipóxia Celular , Modelos Animais de Doenças , Regulação para Baixo/genética , Técnicas de Silenciamento de Genes , Humanos , Injeções Intralesionais , Masculino , Células-Tronco Mesenquimais/citologia , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Infarto do Miocárdio/genética , Infarto do Miocárdio/patologia , Ratos , Regulação para CimaRESUMO
OBJECTIVE: To elucidate the influence of microRNA-409 and the Jak-Stat pathway on the development of liver cancer. PATIENTS AND METHODS: The quantitative Real-time polymerase chain reaction (qRT-PCR) method was used to detect the expression of microRNA-409 in hepatocarcinoma, paracancerous tissues and normal liver tissues, and the correlation between its expression and clinicopathological parameters of patients was analyzed, with the area under the microRNA-409 curve (AUC) being detected. The level of microRNA-409 in different liver cancer cells was detected by qPCR. Then it was overexpressed or knock-downed in the liver cancer cells by cell transfection technique. The cell apoptosis and viability after inhibition or overexpression of microRNA-409 were evaluated by propidium iodide (PI) staining and cell counting kit-8 (CCK-8) assay. Subsequently, Jak2 and Stat3 mRNA levels were detected by qPCR in hepatocarcinoma and paracancerous tissues, with their protein levels analyzed by Western blot after microRNA-409 was inhibited or up-regulated. At last, CCK-8 assay was performed to evaluate the effect of Jak2 on cell viability. RESULTS: Compared with paracancerous and normal liver tissues, the level of microRNA-409 was remarkably reduced in hepatocarcinoma tissues and was negatively correlated with tumor stage, tumor size and overall survival time of patients with liver cancer. Meanwhile, microRNA-409 expression in hepatoma cell lines was also strikingly lower than that in normal liver cells. After overexpression of microRNA-409 in HHCC, cell viability significantly decreased while apoptosis increased, and opposite results were shown in HepG2 cells after miR409 was knock-downed. In liver cancer tissues, the levels of Jak2 and Stat3 were significantly higher than those in adjacent tissues. Additionally, up-regulating microRNA-409 reduced the level of Jak2 and Stat3 protein, while down-regulating it elevated them. In addition, Jak2 could reverse the inhibitory effect of microRNA-409 on the proliferation of hepatoma cells. CONCLUSIONS: Highly-expressed microRNA-409 can down-regulate the Jak-Stat signaling pathway and inhibit cell proliferation to slow down the progression of liver cancer.
Assuntos
Carcinoma Hepatocelular/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/genética , MicroRNAs/metabolismo , Transdução de Sinais/genética , Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/cirurgia , Progressão da Doença , Regulação para Baixo , Feminino , Técnicas de Silenciamento de Genes , Células Hep G2 , Hepatectomia , Humanos , Janus Quinase 2/metabolismo , Fígado/patologia , Fígado/cirurgia , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/cirurgia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Fator de Transcrição STAT3/metabolismo , Análise de SobrevidaRESUMO
Nickelâ»titanium (Ni-Ti) alloy has been selected as stent material given its good biocompatibility. In this study, experimental research on this material was conducted using magnetic field-assisted electrical discharge machining (EDM). The surface topography of the machined workpiece was analyzed with a scanning electron microscope (SEM). Hydrophobicity was measured by using an optical contact angle measuring instrument. The roughness values of different positions on the surface were measured using a TR200 roughness instrument. Results showed that the composite structure of solidification bulgeâ»craterâ»poreâ»particle can be prepared on the surface of the Ni-Ti alloy through magnetic mixed EDM using suitable processing parameters. Moreover, the contact angle of the surface reaches 138.2°.
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OBJECTIVE: The aim of this study was to explore the role of microRNA-210 (miR-210) and E2F3 in the development of pancreatic cancer and to investigate the possible underlying mechanism. PATIENTS AND METHODS: The expression level of miR-210 in pancreatic cancer tissues, para-cancerous tissues, and normal pancreatic tissues was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between miR-210 expression and pathological indicators of pancreatic cancer was analyzed. Meanwhile, the expression of miR-210 in pancreatic cancer cells and normal pancreatic ductal epithelial cells was detected by qRT-PCR. After transfection with miR-210 mimics and inhibitor, the viability and cell cycle of pancreatic cancer cells were detected by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. The binding condition of miR-210 and E2F3 was verified by Dual-Luciferase reporter gene assay. RESULTS: MiR-210 was lowly expressed in pancreatic cancer tissues than that of para-cancerous tissues. The expression of miR-210 was negatively correlated with TNM stage and tumor size of pancreatic cancer. In vitro experiments showed that the miR-210 was downregulated in pancreatic cancer cells than that of normal pancreatic ductal epithelial cells. Meanwhile, overexpression of miR-210 arrested cell cycle decreased cell viability and downregulated E2F3 expression in pancreatic cancer cells. Dual-Luciferase reporter gene assay indicated that E2F3 bound to mi-210. Further experiments confirmed that E2F3 was negatively regulated by miR-210. CONCLUSIONS: MiR-210 knockdown promotes cell proliferation by upregulating E2F3 expression, thereby promoting the progression of pancreatic cancer.
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Fator de Transcrição E2F3/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/metabolismo , Neoplasias Pancreáticas/genética , Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Sobrevivência Celular/genética , Regulação para Baixo , Feminino , Técnicas de Silenciamento de Genes , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Pâncreas/patologia , Neoplasias Pancreáticas/patologiaRESUMO
OBJECTIVE: To explore whether long noncoding RNA (lncRNA) LOC554202 could regulate proliferative and migratory abilities of gastric cancer (GC) cells. MATERIALS AND METHODS: Expression level of LOC554202 in GC cell lines HGC-27 and MGC-803, as well as normal gastric mucosal cell line GES-1 was detected by quantitative real-time polymerase chain reaction (qRT-PCR). LOC554202 knockdown or overexpression in HGC-27 and MGC-803 cells was achieved by transfection of LOC554202-siRNA or pcDNA-LOC554202, respectively. Cell cycle in GC cells was accessed by flow cytometry. Migratory ability of GC cells was determined by wound healing assay and transwell assay. Finally, protein expressions of p21 and E-cadherin in GC cells were detected by Western blot. RESULTS: LOC554202 expression was higher in GC cells than that of gastric mucosal cells (p<0.01). Overexpression of LOC554202 in MGC-803 cells enhanced proliferative and migratory abilities, but decreased protein expressions of p21 and E-cadherin (p<0.01). On the contrary, LOC554202 overexpression in HGC-27 cells decreased proliferative and migratory abilities, but increased protein expressions of p21 and E-cadherin (p<0.01). CONCLUSIONS: LncRNA LOC554202 is highly expressed in GC cells. It could promote proliferative and migratory abilities by downregulating expression levels of p21 and E-cadherin in GC cells.
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Antígenos CD/genética , Caderinas/genética , Inibidor de Quinase Dependente de Ciclina p21/genética , Regulação Neoplásica da Expressão Gênica , RNA Longo não Codificante/metabolismo , Neoplasias Gástricas/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação para Baixo , Técnicas de Silenciamento de Genes , Humanos , Invasividade Neoplásica/genética , RNA Longo não Codificante/genética , RNA Interferente Pequeno/metabolismo , Neoplasias Gástricas/patologiaRESUMO
The AMP-activated protein kinase (AMPK) is a master regulator of metabolic homeostasis. It is activated by the upstream kinase LKB1 (liver kinase B1) when the AMP/ATP ratio is increased during starvation or heightened exercises. Based on reconstitution experiments using purified individual proteins, AMPK was demonstrated to be directly phosphorylated on its conserved residue Thr172 by LKB1, which was promoted by increased levels of AMP. However, recent studies have engendered a paradigm shift for how AMPK is activated inside the cell or animal tissues, unraveling that AXIN binds to LKB1 and tethers it to AMPK located on the surface of late endosome and lysosome (hereafter, only lysosome is discussed) in response to glucose starvation. Moreover, AXIN extends its interaction with the v-ATPase-Ragulator complex, which is paradoxically also required for activation of mTORC1 despite the fact that the two kinases AMPK and mTORC1 are inversely activated. Here, we summarize the experimental procedures of the assays for translocation of AXIN/LKB1 to the detergent-resistant lipid fractions of lysosomal membrane and the assembly of AMPK-activating complexes thereon. These methods will be useful for determining whether AMPK activation induced by various metabolic stresses or by pharmacological stimuli is mediated by the v-ATPase-Ragulator-AXIN/LKB1 axis.
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Proteínas Quinases Ativadas por AMP/análise , Proteínas Quinases Ativadas por AMP/metabolismo , Lisossomos/metabolismo , Biologia Molecular/métodos , Acetiltransferases/genética , Acetiltransferases/isolamento & purificação , Animais , Proteína Axina/genética , Proteína Axina/metabolismo , Células Cultivadas , Ativação Enzimática , Glucose/metabolismo , Imunoprecipitação/métodos , Lisossomos/química , Masculino , Camundongos Endogâmicos C57BL , Fosforilação , Proteínas Serina-Treonina Quinases/metabolismoRESUMO
We aimed to control the gene expression of vascular endothelial growth factor (VEGF) and angiopoietin-1 (Ang-1) in the ischemic heart to explore the feasibility of sequential, timely and controlled multigene expression as a means of improving therapeutic angiogenesis in vivo. Adult rabbit myocardial infarction models were surgically established (n=120). Hypoxia-inducible factor-1α-hypoxic response element (HIF1α-HRE) and Tet (tetracycline)-On advanced gene control systems were reconstructed for controlled expression of the human VEGF165 (hVEGF165) and Ang-1 genes, respectively. Recombinant adeno-associated viruses (rAAV)-9HRE-hVEGF165 and rAAV-TRE-Tight-Ang-1 were delivered into the ischemic myocardium for 12 weeks. Reverse transcription-polymerase chain reaction, western blotting and immunofluorescence staining were used to detect gene and protein expression. Vessel functionality, vascular permeability and animal cardiac function were also evaluated. Under the control of the HIF1α-HRE and Tet-On gene control systems, the expression of the exogenous hVEGF165 and Ang-1 genes was consistent in the ischemia control. In the sequential group, we found that the number of functional vessels with a larger diameter and more vascular branches was increased, and vascular permeability was significantly reduced. In addition, animal heart function was significantly improved compared with the non-sequential and hVEGF165- or Ang-1-only groups (P<0.05, P<0.05, respectively). Sequential, timely and controlled expression of the hVEGF165 and Ang-1 genes in vivo is a new therapeutic angiogenesis strategy that can effectively promote functional vessel regeneration and can improve cardiac function in ischemic heart disease.
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Angiopoietina-1/genética , Angiopoietina-1/metabolismo , Terapia Genética , Infarto do Miocárdio/terapia , Neovascularização Fisiológica , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Permeabilidade Capilar , Vasos Coronários/metabolismo , Vasos Coronários/fisiologia , Dependovirus/genética , Modelos Animais de Doenças , Regulação da Expressão Gênica , Coração/fisiologia , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/biossíntese , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Microvasos/metabolismo , Microvasos/fisiologia , Infarto do Miocárdio/genética , Infarto do Miocárdio/fisiopatologia , Elementos de Resposta , Transdução GenéticaRESUMO
OBJECTIVE: To study the mechanism of 5-aza-2-deoxycytidine (DAC; a methylation inhibitor) on growth of the human cholangiocarcinoma QBC939 cell line. METHODS: A colourimetric assay was used to detect growth of QBC939 cells treated with DAC (0.1-100 µmol/l) over 24 h, 48 h and 72 h. Cell morphology was observed by transmission electron microscopy (TEM). The cell cycle and apoptosis were analysed by flow cytometry. Hypermethylation of the promoters of the p53-BAX mitochondrial apoptosis genes cyclin-dependent kinase inhibitor 2A (CDKN2A), death-associated protein kinase 1 (DAPK1) and PYD and CARD domain containing (PYCARD) was detected by methylation-specific polymerase chain reaction, with and without DAC treatment. RESULTS: DAC inhibited QBC939 cell growth with a half maximal inhibitory concentration of 5 µmol/l at 72 h. After DAC treatment, apoptosis was observed by TEM. Flow cytometric analysis of propidium iodide-positive cells demonstrated increased apoptosis of DAC-treated QBC939 cells (43.04%) compared with untreated cells (4.31%). DAC treatment resulted in demethylation of the gene promoters of CDKN2A and DAPK1 in QBC939 cells. CONCLUSIONS: DAC induces apoptosis of QBC939 cells by reactivation of hypermethylated p53-BAX mitchondrial apoptosis genes in cholangiocarcinoma cells.
Assuntos
Apoptose/genética , Azacitidina/farmacologia , Colangiocarcinoma/patologia , Metilação de DNA/efeitos dos fármacos , Mitocôndrias/genética , Proteína Supressora de Tumor p53/genética , Proteína X Associada a bcl-2/genética , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Colangiocarcinoma/genética , Colangiocarcinoma/ultraestrutura , Metilação de DNA/genética , Humanos , Mitocôndrias/efeitos dos fármacos , Regiões Promotoras Genéticas/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genéticaRESUMO
BACKGROUND: In recent years, many clinical studies have confirmed the value of laparoscopy-assisted gastrectomy (LAG) in gastric cancer surgery, especially in early stages. But the safety and oncologic adequacy of laparoscopy-assisted D2 radical gastrectomy for advanced gastric cancer are still in debate. We conducted a prospective randomized trial to compare open versus laparoscopy-assisted D2 radical gastrectomy in advanced gastric cancer. METHODS: For this study, 123 patients who had been diagnosed endoscopically with gastric cancer were randomly assigned to either LAG (n = 61) or open gastrectomy (OG) (n = 62) which ran from March 2008 to December 2009. Clinical characteristics, operative findings, postoperative recovery, morbidity, pathological report and survival rate were compared. D2 lymph node dissection was performed in 49 patients in the LAG group and 47 patients in the OG group with advanced gastric cancer. We adopt sub-group analysis in this paper. RESULTS: The clinical characteristics of patients in the LAG and OG groups who were in the advanced stage, included age, sex, BMI and concurrent illness, and their ECOG scores were well matched. Operative findings, postoperative recovery, morbidity, pathological findings including tumor location, depth of invasion, TNM stage, histological grade and surgical extension in the two groups were also similar. Compared to the OG group, the mean operating time was significantly longer for the LAG group (267.88 ± 54.284 min in the LAG group vs. 182.02 ± 41.016 min in the OG group, p = 6.383 × 10(-13)); the mean number of days when body temperature exceeded 37°C was significantly shorter in the LAG group (p = 6.34 × 10(-8)). There were no postoperative deaths in both the groups. The postoperative morbidity rate was 12.24% in the LAG group and 19.15% in the OG group with no significant difference (p = 0.357). However, pulmonary infection was observed more frequently in the OG group (p = 0.038). After a mean follow-up of 22.1354 months (from 4 to 36 months), 14 and 15 patients died of gastric cancer in the LAG and OG groups, respectively. Two and one patient died of nongastric cancer in the LAG and OG groups, respectively. The overall survival rates were 67.1% and 53.8% in the LAG and OG groups, respectively. The estimated mean survival time was 29.387 months in the LAG group and 28.978 months in the OG group. There was no statistically significant difference in the overall survival rate for patients in both groups - LAG and OG (log-rank test, p = 0.911, Tarone Ware test, p = 0.994, and Breslow test, p = 0. 961). CONCLUSION: LAG with D2 lymph node dissection is a safe and feasible procedure with adequate lymphadenectomy, good curability and survival rate for the treatment of advanced gastric cancer.
Assuntos
Adenocarcinoma/patologia , Adenocarcinoma/cirurgia , Gastrectomia/métodos , Excisão de Linfonodo , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgia , Cavidade Abdominal , Idoso , Feminino , Febre/etiologia , Gastrectomia/efeitos adversos , Humanos , Estimativa de Kaplan-Meier , Laparoscopia/efeitos adversos , Excisão de Linfonodo/efeitos adversos , Masculino , Pessoa de Meia-Idade , Fatores de TempoRESUMO
Influenza is a topic of wide public concern, particularly because of the recent emergence of avian flu. The myxovirus resistance (Mx) protein has been shown to have an inhibitory effect on influenza virus and is therefore of great interest. This study examines the Mx protein in 8 local Chinese chicken breeds and 2 exotic chicken breeds. Amino acid 631, found in the Mx GTPase effector domain, was examined in 534 individuals by comparing PCR results, and individuals were separated into the A/A genotype or the G/G genotype, depending on whether amino acid 631 is an Asn or Ser. In the native breed, the frequency of G/G homozygotes is 0.780 (294/377). The Mx expression levels in tissues and chicken embryo fibroblast cells with different genotypes were also studied. The A/A individuals from Beijing-You and White Leghorn breeds had higher Mx expression levels than G/G individuals. The liver, heart, and spleen had higher expression levels than muscle or kidney. The A/A chicken embryo fibroblast cells had higher antiviral activity against vesicular stomatitis virus and Newcastle disease. We provide the first report examining the expression level and antiviral activity of different Mx alleles of nucleotide 2216(S631N) genotypes. This study lays a good foundation for correlative studies examining genotype and antiviral function.
Assuntos
Substituição de Aminoácidos , Galinhas/genética , Proteínas de Ligação ao GTP/genética , Regulação da Expressão Gênica/imunologia , Orthomyxoviridae/fisiologia , Animais , Células Cultivadas , Fibroblastos/efeitos dos fármacos , Proteínas de Ligação ao GTP/metabolismo , Proteínas de Ligação ao GTP/farmacologia , Predisposição Genética para Doença , Genótipo , Proteínas de Resistência a Myxovirus , Infecções por Orthomyxoviridae/genética , Infecções por Orthomyxoviridae/veterinária , Doenças das Aves Domésticas/genética , Doenças das Aves Domésticas/virologia , RNA Mensageiro , RNA ViralRESUMO
BACKGROUND: Allergic rhinitis affects 10-40% of the population globally with a substantial health and economic impact on the community. OBJECTIVE OF REVIEW: To assess the effectiveness and safety of ear-acupuncture or ear-acupressure for the treatment of allergic rhinitis by reviewing randomised controlled trials and quasi-randomised controlled trials. TYPE OF REVIEW: This review followed the methods specified in the Cochrane Handbook for Systematic Reviews of Interventions. SEARCH STRATEGY: A total of 21 electronic English and Chinese databases were searched from their respective inceptions to April 2008. Key words used in the search included the combination of ear, auricular, acupuncture, acupressure, acupoint, allergic, allergy, rhinitis, hayfever, randomised clinical trial and their synonyms. EVALUATION METHOD: The methodological quality was assessed using Jadad's scale. The effect size analysis was performed to explore the difference between interventional groups. RESULTS: Ninety-two research papers were identified and seven of them referring to five studies met the inclusion criteria. All included studies involved ear-acupressure treatment. These studies mentioned randomisation, but no details were given. None of the five studies used blinding or intention-to-treat analysis. Ear-acupressure was more effective than herbal medicine, as effective as body acupuncture or antihistamine for short-term effect, but it was more effective than anti-histamine for long-term effect. CONCLUSIONS: The benefit of ear-acupressure for symptomatic relief of allergic rhinitis is unknown due to the poor quality of included studies.
Assuntos
Acupressão/métodos , Orelha/fisiologia , Rinite Alérgica Perene/fisiopatologia , Rinite Alérgica Perene/terapia , HumanosRESUMO
BACKGROUND: Omentum is well known for its immunocompetence and good blood supply; and therefore, is being used in various complex thoracic procedures. Specially, in situations when staplers, sealants and total parenteral nutrition may not be used because of financial constraints, omentum may prove very helpful in preventing post-operative fatal complications. METHODS: A retrospective review of 61 patients was undertaken. Patients were categorized into two groups. In group I, omentum was sutured to the anastomosis for prophylaxis of leak from gastro-oesophagectomy after radical surgery for cancer of cardia and oesophagus. In group II, it was used for therapeutic purpose, to control diffuse air leak from lung parenchyma after chest wall and invaded lung resection for malignant chest wall tumours (subgroup A) and treatment of post pneumonectomy bronchopleural fistula for NSCLC of right lung (subgroup B). Gastro-oesophagectomy, closure of bronchial stump and suturing of lung parenchyma after wedge resection was done with manual suturing technique only. RESULTS: Group I: There were 57 patients with the diagnosis of cancer of cardia and oesophagus, who underwent radical surgery. Transthoracic approach was used in 96.5% patients, whereas 3.5% patients underwent transhiatal resection. Anastomotic level was located in chest in 68.4% and in neck in 31.6% patients. The leakage rate was 5.4%. Group II: There were three patients in subgroup A, all with lesions located in left side of chest wall. There was one patient in subgroup B. Chest tube was removed after a mean time of 2 days and after 4 days in subgroup A and B, respectively. There was 1 mortality (1.6%) secondary to chylothorax. CONCLUSION: Use of pedicled omentum appears to be a very simple technique to prevent the anastomotic leak after radical surgery for cancer of cardia and oesophagus, and to seal the diffuse parenchymal pulmonary leak after various procedures in thorax.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/cirurgia , Neoplasias Esofágicas/cirurgia , Neoplasias Pulmonares/cirurgia , Omento/transplante , Neoplasias Gástricas/cirurgia , Anastomose Cirúrgica , Cárdia/cirurgia , Esofagectomia , Feminino , Gastrostomia , Humanos , Masculino , Pessoa de Meia-Idade , Omento/irrigação sanguínea , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/prevenção & controle , Estudos Retrospectivos , Parede Torácica/cirurgia , Resultado do TratamentoRESUMO
A survey was done in 15 typical villages, 150 soil and 86 vegetable plant samples were taken in Jiaxin prefecture of the Taihu Lake region, northern Zhejian province. Results indicate that after 15-20 years land use changed from the paddy rice-wheat (or oilseed rape) double cropping system, to a continuous vegetable land has caused soil quality dramatic change. (1) Acidification: average soil pH was 5.4; about 61% of total samples were pH < 5.5. It was 0.9 units lower than 10 years ago with same upland vegetable cultivation and was 1.2 units lower than soil pH of paddy rice-wheat (or oilseed rape) rotation. (2) Fertilizer salt accumulation: the average salt content was 0.28%, among these about 36.2% of the total samples contained more than 0.3%. (3) Nitrate N and available phosphorus (P) over accumulation: on average it was 279 mg NO3-N/kg, and 45-115 mg P/kg. Nitrate N four times higher and available P 4-10 times more than it is in present paddy rice-wheat rotation soils respectively. This has caused wide concern because of possible groundwater and well drinking water pollution by leached nitrate N and the P losses to water by runoff from vegetable lands induce surface water eutrophication.