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INTRODUCTION: Organ preservation is now considered an acceptable alternative option in distal rectal cancer patients with clinical complete response (cCR) after neoadjuvant chemoradiation (CRT). But the cCR rate is low and about one-third of tumour will regrow, which requires more effective local treatment. CRT combined with intra-arterial chemotherapy (IAC) might be a promising approach. Additionally, total neoadjuvant therapy using FOLFIRINOX induction chemotherapy improved survival while consolidation chemotherapy improved organ preservation. We assess whether IAC plus CRT and FOLFIRINOX consolidation chemotherapy can improve the chance of organ preservation and survival in distal rectal cancer. METHODS AND ANALYSIS: This prospective, monocentric, open-label, single-arm phase II study will include 32 patients with cT3-4NanyM0 distal rectal adenocarcinoma. All patients will receive one cycle of IAC (irinotecan, raltitrexed and oxaliplatin), followed by CRT (50 Gy/25 fractions with concomitant capecitabine) and then with six cycles of FOLFIRINOX (leucovorin, 5-fluorouracil, oxaliplatin and irinotecan). After final evaluation, patients with cCR will receive non-operative management or surgery at their own discretion and others are mandatorily referred to surgery. Adjuvant chemotherapy with six cycles of mFOLFOX6 (leucovorin, 5-fluorouracil and oxaliplatin) will be used for patients with adverse pathological features. The primary endpoint is the rate of complete response (CR; pathological CR or sustained cCR≥2 years). The main secondary endpoints are toxicity, compliance, short-term and long-term oncological outcomes, surgical morbidity and quality of life. This protocol has been designed in accordance with the Standard Protocol Items: Recommendations for Interventional Trials 2013 guidelines. ETHICS AND DISSEMINATION: This study was approved by the Academic and Ethics Committee of The Affiliated Hospital of Youjiang Medical University for Nationalities in March 2023. Trial results will be published in peer-reviewed international journals and on the ChiCTR website. PROTOCOL VERSION: Registered on 18 April 2023; version #1. TRIAL REGISTRATION NUMBER: ChiCTR2300070620.
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Neoplasias Pancreáticas , Neoplasias Retais , Humanos , Terapia Neoadjuvante/métodos , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Irinotecano/uso terapêutico , Oxaliplatina/uso terapêutico , Leucovorina/uso terapêutico , Qualidade de Vida , Estudos Prospectivos , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Retais/tratamento farmacológico , Neoplasias Retais/patologia , Fluoruracila/uso terapêutico , Quimiorradioterapia/métodos , Estadiamento de Neoplasias , Ensaios Clínicos Fase II como AssuntoRESUMO
Background: Lung cancer is one of the most common malignant tumors in the world. Exportins are closely associated with the cellular activity and disease progression in a variety of different tumors. However, the expression level, genetic variation, immune infiltration, and biological function of different exportins in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC), as well as their relationship with the prognosis of patients with LUAD and LUSC have not been fully clarified. Methods: To analyze the differential expression, prognostic value, genetic variation, biological function, and immune cell infiltration of exportins in patients with LUAD and LUSC, the ONCOMINE; UALCAN; Human Protein Atlas (HPA); Kaplan-Meier plotter; cBioPortal; Search Tool for the Retrieval of Interacting Genes/Proteins (STRING); Database for Annotation, Visualization, and Integrated Discovery (DAVID); Tumor Immune Estimation Resource (TIMER); and LinkedOmics databases were used in this study. Results: The transcriptional and protein expression levels of CSE1L and XPO1/5/6/7 were increased in patients with LUAD and LUSC, and the increased transcriptional levels of CSE1L and XPO5/6/7 were related to worse prognosis. An increased transcriptional level of XPO1 was associated with a better prognosis. These results indicated that CSE1L and XPO1/5/6/7 may be potential prognostic biomarkers for the survival of patients with LUAD and LUSC. Moreover, the high mutation rate of exportins in non-small cell lung cancer was 50.48%, and the largest proportion of mutations included high messenger RNA expression. The expression of exportins was significantly correlated with the infiltration of various immune cells. Differentially expressed exportins could regulate the occurrence and development of LUAD and LUSC by involving a variety of microRNAs and transcription factor E2F1. Conclusions: Our study provides novel insights into the selection of prognostic biomarkers of exportins in LUAD and LUSC.
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LINC00511 is an long non-coding RNA (lncRNA) of ncRNAs,This study aimed to investigate whether the lncRNA LINC00511 could encode a small peptide, LINC00511-133aa, and whether this peptide could promote breast cancer cell metastasis and stemness by activating the wnt/ß-catenin pathway. The LINC00511-133aa coding sequence vector and control vector were transfected into MCF-7 and MDA-MB-231 breast cancer cells, with subsequent assessment of peptide expression using PCR, western blotting, and immunofluorescence assays. Cell proliferation, invasion, and apoptosis were evaluated using CCK8, apoptotic, wound healing, and transwell invasion assays, while the characteristic changes of tumor stem cells were detected through sphere-forming assay and western blot analyses of the stemness markers Oct4, Nanog, and SOX2. Results showed that LINC00511-133aa was indeed encoded by LINC00511 and promoted the invasiveness and stemness of breast cancer cells while limiting apoptosis by modulating the expression levels of wnt/ß-catenin pathway-related proteins Bax, c-myc, and CyclinD1, as well as facilitating ß-catenin protein entry into the nucleus. This study provides evidence for the potential involvement of lncRNA LINC00511 and its peptide product in breast cancer progression via the regulation of the wnt/ß-catenin pathway.
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Neoplasias da Mama , RNA Longo não Codificante , Humanos , Feminino , beta Catenina/genética , beta Catenina/metabolismo , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , RNA Longo não Codificante/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Peptídeos/genética , Peptídeos/metabolismo , Invasividade Neoplásica/genéticaRESUMO
Resistance to chemotherapeutic drugs limits the efficacy of chemotherapy in non-small cell lung cancer (NSCLC). Autophagy is an essential mechanism which involves in drug resistance. Our previous research has revealed that miR-152-3p represses NSCLC progression. However, the mechanism of miR-152-3p in autophagy-mediated chemoresistance in NSCLC remains unclear. Cisplatin-resistant cell lines (A549/DDP and H446/DDP) were transfected with related vectors and subjected to cisplatin, autophagy inhibitor, activator, or extracellular signal-regulated kinase (ERK) activator. Flow cytometry, CCK8 and colony formation assays were performed for testing apoptosis and cell viability. The related RNAs or proteins were detected by qRT-PCR or Western blot. Chromatin immunoprecipitation, luciferase reporter assay or RNA immunoprecipitation were used for validating the interaction between miR-152-3p and ELF1 or NCAM1. Co-IP verified the binding between NCAM1 and ERK. The role of miR-152-3p in cisplatin resistance of NSCLC was also validated in vivo. The results showed that miR-152-3p and ELF1 were decreased in NSCLC tissues. miR-152-3p reversed cisplatin resistance by inhibiting autophagy through NCAM1. NCAM1 promoted autophagy through the ERK pathway and facilitated cisplatin resistance. ELF1 positively regulated miR-152-3p level by directly interacting with miR-152-3p promoter. miR-152-3p targeted NCAM1 to regulate NCAM1 level and then affected the binding of NCAM1 to ERK1/2. ELF1 inhibited autophagy and reversed cisplatin resistance through miR-152-3p/NCAM1. miR-152-3p inhibited autophagy and cisplatin resistance of xenograft tumor in mice. In conclusion, our study revealed that ELF1 inhibited autophagy to attenuate cisplatin resistance through the miR-152-3p/NCAM1/ERK pathway in H446/DDP and A549/DDP cells, suggesting a potential novel treatment strategy for NSCLC.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , MicroRNAs , Animais , Humanos , Camundongos , Autofagia/genética , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Antígeno CD56 , Linhagem Celular Tumoral , Proliferação de Células/genética , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Resistencia a Medicamentos Antineoplásicos/genética , MAP Quinases Reguladas por Sinal Extracelular , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/genética , MicroRNAs/metabolismo , Proteínas Nucleares , Fatores de Transcrição/genéticaRESUMO
BACKGROUND: Systematic lupus erythematosus (SLE) is an autoimmunemediated disease. So far, there is no relevant report on ferroptosis in SLE research, and the role of T helper 1 (Th1) and T helper 2 (Th2) cells in SLE is still unclear. METHODS: This study employed SLE mice models with and without ferroptosis inhibitors (Liproxstatin1) and normal control mice. Treated mice were analyzed with hematoxylin and eosin (H&E) staining, immunohistochemical detection of glutathione peroxidase 4 (GPX4), malondialdehyde (MDA) detection, ELISA(enzyme-linked immunosorbent assay) detection of Th1 and Th2 cytokines and flow cytometry detection of Th1 and Th2 ratio. RESULTS: The results showed that compared with the normal group, the SLE group exhibited significantly higher expression of anti-double-stranded deoxyribonucleic acid (anti-dsDNA), MDA and Th1 cytokines, significantly lower expression of GPX4 and Th2 cytokines and increased Th1/Th2 ratio. Similarly, compared with the SLE group, the SLE + liproxstatin-1 group showed significantly low expression of anti-dsDNA, MDA and Th1 cytokines, significantly high expression of GPX4 and Th2 cytokines and reduced Th1/Th2 ratio. CONCLUSION: These results demonstrate that ferroptosis may be involved in promoting SLE development. Therefore, inhibiting ferroptosis may be a potential treatment for SLE. Similarly, the Th1/Th2 ratio may have a role in promoting SLE development.
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Ferroptose , Lúpus Eritematoso Sistêmico , Animais , Camundongos , Células Th1 , Citocinas/metabolismo , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Células Th2/metabolismo , Progressão da DoençaRESUMO
Background: Long non-coding RNA (lncRNA) regulates the tumorigenesis as well as the development of lung adenocarcinoma (LUAD), which is one of the high-mortality cancers. We explored the influence of lncRNA AC098934 on the malignant biological behavior of LUAD and potential underlying molecular mechanisms. Methods: The expression level of AC098934 in either the LUAD or the normal tissues was identified in the TCGA database. Two AC098934 knockdown siRNAs were infected into cells of LUAD, including A549 as well as H1299 cells, using the lentivirus. Real-time Quantitative polymerase chain reaction (QPCR) helped to determine the knockdown efficiency of AC098934. CCK-8, cell cloning, wound healing combined with transwell assays tested the role of AC098934 in the cell proliferation, migration as well as the invasion. Tumor formation experiment in nude mice subcutaneously confirmed the promoting effect of AC098934 in vivo. In addition, combinations of METTL3 and AC098934, as well as m6A and AC098934 were identified through the RIP assay. Results: Compared to the normal tissues, AC098934 was more highly expressed in LUAD tissues. After AC098934 was knocked down by siRNA, the proliferation, invasion, migration as well as tumorigenesis abilities of both A549 and H1299 cells were reduced. Mechanistically, AC098934 could bind to the m6A antibody and METTL3 protein. METTL3 overexpression promoted the m6A modification on AC098934, thereby increasing the interaction of m6A modification. Conclusion: The highly expressed lncRNA AC098934 in LUAD facilitates the cell proliferation as well as invasion either in vitro or in vivo. METTL3 binds, furthermore, modulates the m6A modification of AC098934. Our research revealed a new molecular mechanism, through which AC098934 promoted the malignant behavior of LUAD tumors under the m6A modification induced by METTL3. This indicates that AC098934 is possible to be a promising biomarker as well as a therapeutic target for the patients with LUAD.
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The implications of the CD40-CD40 ligand (CD40L) signaling pathway in systemic lupus erythematosus (SLE) were well documented, due to its important role among immune cells. Previous research found that 3-hydroxy butyrate dehydrogenase 2 (BDH2), a modulator of intracellular iron homeostasis and iron transportation promoted the pathogenic process of SLE by regulating the demethylation of cd70, cd11a, and cd40l genes among CD4 + T cells. The purpose of this study was to explore the role of BDH2 in oxidative damage-induced SLE. First, CD4 + T cells treated with H2O2 were injected into the tail vein of mice to establish a lupus model. CD40L knockdown significantly decreased CD40L expression on CD4 + T cells in the spleen of SLE mice. Compared with SLE model mice, the levels of serum anti-dsDNA antibody and urinary protein in the CD40L interference group were significantly decreased. CD40L knockdown alleviated the immune complex glomerulonephritis in syngeneic SLE mice. Moreover, the levels of IFN-γ and IL-2 were decreased. However, IL-4 and IL-10 levels were significantly upregulated in the serum of CD40L knockdown SLE mice, compared with SLE model mice. Accordingly, CD40L knockdown reduced Th1/Th2 percentage in SLE mice. Inhibiting the expression of BDH2 of CD4 + T cells promoted the demethylation of CD40L, while it inhibited cell proliferation, elevated oxidative stress through increased expression of CD40L, and thus, promoted the progress of SLE. Our results demonstrate that BDH2 aggravates the pathologic progression of SLE in mice, by increasing the demethylation level of CD40L among CD4 + T cells.
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Ligante de CD40/imunologia , Hidroxibutirato Desidrogenase/deficiência , Lúpus Eritematoso Sistêmico/imunologia , Células Th1/imunologia , Células Th2/imunologia , Animais , Ligante de CD40/genética , Modelos Animais de Doenças , Feminino , Hidroxibutirato Desidrogenase/imunologia , Lúpus Eritematoso Sistêmico/genética , Metilação , Camundongos , Camundongos Endogâmicos BALB CRESUMO
The purpose of the present study was to examine the epigenetic mechanism by which microRNA (miR)-152-3p regulates proliferation in non-small cell lung cancer A549 cells via neural cell adhesion molecule 1 (NCAM1). Bisulfite sequencing PCR (BSP), the gold standard for methylation detection, uses bisulfite-treated DNA to determine its pattern of methylation. Treatment of DNA with bisulfite converts cytosine residues to uracil, but leaves 5-methylcytosine residues unaffected. It was conducted and demonstrated a relatively high level of methylation in the miR-152-3p promoter region. Chromatin immunoprecipitation was combined with PCR to detect the binding of DNA methyltransferase 3B (DNMT3B) protein to miR-152-3p, which tends to occur in the core region of the miR-152-3p gene in A549 cells. Luciferase assay identified NCAM1 as the target gene of miR-152-3p. MTT, colony formation and Transwell assays indicated that miR-152-3p could decrease cell proliferation and invasion and in addition to reducing the expression level of NCAM1. Overexpression of NCAM1 could attenuate the effect of miR-152-3p. DNMT3B knockdown decreased the proliferative ability of A549 cells and increased the expression of miR-152-3p, while decreased that of NCAM1. After treatment with miR-152-3p inhibitor, these effects were attenuated and the NCAM1 expression level was upregulated. The results indicated that miR-152-3p may suppress the proliferation of A549 cells by downregulating NCAM1. In addition, DNMT3B negatively regulated the expression of miR-152-3p via modulation of the methylation level in the miR-152-3p core region, thus mediating the proliferation of lung tumor cells.
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Reactive oxygen species, particularly hydrogen peroxide (H2O2), can induce proliferation inhibition and death of A549 cells via oxidative stress. Oxidative stress has effect on DNA methylation. Oxidative stress and DNA methylation feature a common denominator: The one carbon cycle. To explore the inhibitory mechanism of H2O2 on the proliferation of lung cancer cells, the present study analysed the mRNA expression and methylation profiles in A549 cells treated with H2O2 for 24 h, as adenocarcinoma is the most common pathological type of lung cancer. The DNA methylation profile was constructed using reduced representation bisulphite sequencing, which identified 29,755 differentially methylated sites (15,365 upregulated and 14,390 downregulated), and 1,575 differentially methylated regions located in the gene promoters were identified using the methylKit. Analysis of the assocaition between gene expression and methylation levels revealed that several genes were downregulated and hypermethylated, including cyclin-dependent kinase inhibitor 3, denticleless E3 ubiquitin protein ligase homolog, centromere protein (CENP)F, kinesin family member (KIF)20A, CENPA, KIF11, PCNA clamp-associated factor and GINS complex subunit 2, which may be involved in the inhibitory process of H2O2 on the proliferation of A549 cells.
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Lung adenocarcinoma (LUAD) is the most common type of non-small cell lung cancer and has a poor 5 year survival rate (<10%). Cisplatin is one of the most effective chemotherapeutic treatments for LUAD, even though it is of limited overall utility due to acquired drug resistance. To identify possible genetic targets for the mitigation of cisplatin resistance, gene expression data from cisplatin-resistant cell lines were integrated with patient information. Expression data for cisplatin-resistant and cisplatin-sensitive A549 cell lines were obtained from the Gene Expression Omnibus database, while LUAD patient data was obtained from The Cancer Genome Atlas (TCGA) database. Differentially expressed mRNAs (DEmRNAs), microRNAs (DEmiRNAs) and long non-coding RNAs (DElncRNAs) were identified between the cisplatin-sensitive and cisplatin-resistant cells. Using the TCGA patient data, 33 DEmRNAs associated with survival were identified. A total of 74 DElncRNAs co-expressed with the survival-associated DEmRNAs, and 11 DEmiRNAs that regulated the survival-associated DEmRNAs, were also identified. A competing endogenous RNA (ceRNA) network was constructed based on the aforementioned results, which included 17 survival-associated DEmRNAs, 9 DEmiRNAs and 16 DElncRNAs. This network revealed 8 ceRNA pathway axes possibly associated with cisplatin resistance in A549 cells. Specifically, the network suggested that the lncRNAs HOXD-AS2, LINC01123 and FIRRE may act as ceRNAs to increase cisplatin resistance in human LUAD cells. Therefore, it was speculated that these lncRNAs represent potentially rewarding research targets.
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OBJECTIVE: To get a baseline of 32 provincial center for disease control and prevention( CDC) microbiology laboratory in the aspect of quantitative test ability of Staphylococcus aureu, qualitative test ability of diarrheagenic Escherichia coli and unknown intestinal pathogen, and to comprehend the quality of microbiology testing. METHODS: Two different concentration samples of Staphylococcus aureus I and II were made. Diarrheagenic Escherichia coli and unknown intestinal pathogen samples were transported using the special semi-solid AGAR transport medium, in the form of four pure different serotypes each. The results of Staphylococcus aureu were logarithmic transformed, and evaluated with Z-score method using the average and standard deviation. The reporting results of diarrheagenic Escherichia coli and unknown intestinal pathogen were evaluated as right or wrong, and non-reporting were evaluated as missing. RESULTS: The satisfaction rate of Staphylococcus aureus was 82. 81%. The accuracy rate of diarrheagenic Escherichia coli was 45. 16%. Of the 32 results of unknown intestinal pathogen, 29 CDCs reported correctSalmonella serotypes, and the accuracy rate was 90. 63%. Combined the three quality control results, 9 CDCs reached the accurate results, with the accuracy rate of 28. 13%. CONCLUSION: 32 provincial CDCs have the identification ability of Salmonella serotyping and qualitative test ability of Staphylococcus aureu, while they need to strength diarrheagenic Escherichia coli identification and serotyping ability.
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Infecções por Escherichia coli/microbiologia , Escherichia coli/classificação , Escherichia coli/patogenicidade , Laboratórios/normas , Controle de Qualidade , China , Humanos , SorotipagemRESUMO
Lupus develops when genetically predisposed people encounter environmental agents such as UV light, silica, infections and cigarette smoke that cause oxidative stress, but how oxidative damage modifies the immune system to cause lupus flares is unknown. We previously showed that oxidizing agents decreased ERK pathway signaling in human T cells, decreased DNA methyltransferase 1 and caused demethylation and overexpression of genes similar to those from patients with active lupus. The current study tested whether oxidant-treated T cells can induce lupus in mice. We adoptively transferred CD4(+) T cells treated in vitro with oxidants hydrogen peroxide or nitric oxide or the demethylating agent 5-azacytidine into syngeneic mice and studied the development and severity of lupus in the recipients. Disease severity was assessed by measuring anti-dsDNA antibodies, proteinuria, hematuria and by histopathology of kidney tissues. The effect of the oxidants on expression of CD40L, CD70, KirL1 and DNMT1 genes and CD40L protein in the treated CD4(+) T cells was assessed by Q-RT-PCR and flow cytometry. H2O2 and ONOO(-) decreased Dnmt1 expression in CD4(+) T cells and caused the upregulation of genes known to be suppressed by DNA methylation in patients with lupus and animal models of SLE. Adoptive transfer of oxidant-treated CD4(+) T cells into syngeneic recipients resulted in the induction of anti-dsDNA antibody and glomerulonephritis. The results show that oxidative stress may contribute to lupus disease by inhibiting ERK pathway signaling in T cells leading to DNA demethylation, upregulation of immune genes and autoreactivity.
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Autoimunidade/genética , Autoimunidade/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Epigênese Genética , Estresse Oxidativo/genética , Estresse Oxidativo/imunologia , Animais , Linfócitos T CD4-Positivos/efeitos dos fármacos , Metilação de DNA , Modelos Animais de Doenças , Epigênese Genética/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Glomerulonefrite/genética , Glomerulonefrite/imunologia , Glomerulonefrite/patologia , Lúpus Eritematoso Sistêmico/genética , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/patologia , Camundongos , Oxidantes/farmacologiaRESUMO
OBJECTIVE: To test the aerobic plate count examining capability of microbiology laboratories, to ensure the accuracy and comparability of quantitative bacteria examination results, and to improve the quality of monitoring. METHODS: The 4 different concentration aerobic plate count piece samples were prepared and noted as I, II, III and IV. After homogeneity and stability tests, the samples were delivered to monitoring institutions. The results of I, II, III samples were logarithmic transformed, and evaluated with Z-score method using the robust average and standard deviation. The results of IV samples were evaluated as "satisfactory" when reported as < 10 CFU/piece or as "not satisfactory" otherwise. Pearson χ2 test was used to analyze the ratio results. RESULTS: 309 monitoring institutions, which was 99.04% of the total number, reported their results. 271 institutions reported a satisfactory result, and the satisfactory rate was 87.70%. There was no statistical difference in satisfactory rates of I, II and III samples which were 81.52%, 88.30% and 91.40% respectively. The satisfactory rate of IV samples was 93.33%. There was no statistical difference in satisfactory rates between provincial and municipal CDC. CONCLUSION: The quality control program has provided scientific data that the aerobic plate count capability of the laboratories meets the requirements of monitoring tasks.
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Bactérias/isolamento & purificação , Técnicas de Laboratório Clínico/normas , Contagem de Colônia Microbiana/métodos , Controle de Qualidade , Bactérias/crescimento & desenvolvimento , China , Contagem de Colônia Microbiana/normas , Laboratórios/normasRESUMO
OBJECTIVE: Lupus develops when genetically predisposed people encounter environmental agents, such as ultraviolet light, silica, infections, and cigarette smoke, that cause oxidative stress, but how oxidative damage modifies the immune system to cause lupus flares is unknown. We previously showed that inhibiting DNA methylation in CD4+ T cells by blocking ERK pathway signaling is sufficient to alter gene expression, and that the modified cells cause lupus-like autoimmunity in mice. We also reported that T cells from patients with active lupus have decreased ERK pathway signaling, have decreased DNA methylation, and overexpress genes normally suppressed by DNA methylation. This study was undertaken to test whether oxidizing agents decrease ERK pathway signaling in T cells, decrease DNA methyltransferase levels, and cause demethylation and overexpression of T cell genes similar to that found in T cells from patients with active lupus. METHODS: CD4+ T cells were treated with the oxidizers H2 O2 or ONOO(-) . Effects on ERK pathway signaling were measured by immunoblotting, DNA methyltransferase 1 (DNMT-1) levels were measured by reverse transcriptase-polymerase chain reaction (RT-PCR), and the methylation and expression of T cell genes were measured using flow cytometry, RT-PCR, and bisulfite sequencing. RESULTS: H2 O2 and ONOO(-) inhibited ERK pathway signaling in T cells by inhibiting the upstream regulator protein kinase Cδ, decreased DNMT-1 levels, and caused demethylation and overexpression of genes previously shown to be suppressed by DNA methylation in T cells from patients with active lupus. CONCLUSION: Our findings indicate that oxidative stress may contribute to human lupus flares by inhibiting ERK pathway signaling in T cells to decrease DNMT-1 and cause DNA demethylation.
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Linfócitos T CD4-Positivos/metabolismo , Metilação de DNA/fisiologia , Lúpus Eritematoso Sistêmico/fisiopatologia , Estresse Oxidativo/fisiologia , Adulto , Linfócitos T CD4-Positivos/patologia , Células Cultivadas , DNA (Citosina-5-)-Metiltransferase 1 , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA/efeitos dos fármacos , Feminino , Humanos , Peróxido de Hidrogênio/farmacologia , Técnicas In Vitro , Lúpus Eritematoso Sistêmico/metabolismo , Lúpus Eritematoso Sistêmico/patologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Masculino , Pessoa de Meia-Idade , Ácido Peroxinitroso/farmacologia , Proteína Quinase C-delta/metabolismoRESUMO
OBJECTIVE: To find out and compare the professionals proficiency of monitoring and inspection agencies on pesticide residue in food, and identify the main factors affecting the quality of result to provide references for improving the quality of the monitoring and inspection results. METHODS: Compare the national quality control examination results of pesticides residues in food of 2009 with 2011. RESULTS: The submission rates of examination results in 2011 were much higher than in 2009. They were respectively 100% and 83.6%. With the exception of few pesticides, the qualitative qualification rate and quantitative satisfaction rate of other pesticides were also relatively higher, and the qualitative qualification rate and quantitative satisfaction rate in 2011 were all higher than the same examination pesticides in 2009. For instance, the qualitative qualification rate of chlorpyrifos were respectively 99.4% and 96.8% in 2011 and 2009. The qualitative qualification rate of triazophos were respectively 100% and 97.1% in 2011 and 2009. The quantitative satisfaction rate of chlorpyrifos were respectively 93.8% and 90.2% in 2011 and 2009. The quantitative satisfaction rate of triazophos were respectively 91.9% and 87.2% in 2011 and 2009. CONCLUSION: The general testing levels of monitoring and inspection agencies on pesticide residue in food were sufficient, and their testing proficiency increased year by year, but some institutions still need to improve the testing proficiency.
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Análise de Alimentos/normas , Contaminação de Alimentos/estatística & dados numéricos , Resíduos de Praguicidas/análise , China , Contaminação de Alimentos/análise , Humanos , Controle de Qualidade , Medição de RiscoRESUMO
OBJECTIVE: To find out the proficiency of professionals in monitoring and inspection agencies on the determination of pesticides residue in food, and to improve the accuracy and comparability of testing results. METHODS: Three sets of quality control samples with the same matrix but different in concentrations of pesticides were prepared; one of blind quality control samples contained chlorpyrifos, methyl parathion, methyl pirimiphos, fenitrothion or triazophos and bifenthrin was dispatched to each inspection agency. Criteria for testing results were evaluated by Z scores: [Z] = < or = 2 for satisfactory; 2 < [Z] < 3 for uncertainty and [Z] > or = 3 for unsatisfactory. RESULTS: The results of all quality control blind samples delivered to 162 agencies were submitted back on time. The qualitative and quantitative qualification rate for chlorpyrifos was 99.4% and 93.8%, for parathion-methyl was 96.3% and 94.9%, for bifenthrin was 95.7% and 89.7%, for fenitrothion was 92.0% and 89.6%, for triazophos was 100% and 91.9%, respectively. CONCLUSION: The general proficiency on tested pesticide residues in food was relatively good for monitoring and inspection agencies. The agency who got unsatisfied results has recognized the problems in the process of testing; the cause of uncertainty and unsatisfactory results were found; and effective corrections were conducted to ensure the accuracy of data in the future.
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Contaminação de Alimentos , Inocuidade dos Alimentos , Resíduos de Praguicidas/análise , Alimentos , Humanos , Compostos Organotiofosforados , Praguicidas , Controle de Qualidade , Medição de RiscoRESUMO
AIM: To investigate the apoptosis-inducing effect of trichostatin A (TSA) in the human lung adenocarcinoma cisplatin-resistant cell line (A549/CDDP) and to examine whether TSA can enhance sensitivity to cisplatin treatment and the underlying molecular mechanisms of such an enhancement. METHODS: Cell viability was evaluated using the Neutral Red assay. Apoptosis was assessed using Hoechst 33258 staining and flow cytometry analysis. Protein expression was detected by Western blotting. To determine the role of Death-associated protein kinase (DAPK) in TSA-induced apoptosis in the A549/CDDP cell line, cells were transfected with pcDNA3.1(+)-DAPK, which has a higher expression level of DAPK compared to endogenous expression, and DAPK activity was inhibited by both over-expression C-terminal fragment of DAPK which may competitive binding DAPK substrates to inhibit the function of DAPK and RNA interference. RESULTS: TSA induced apoptosis in both A549 cells and A549/CDDP cells. TSA enhanced the sensitivity of A549/CDDP cells to cisplatin, along with concomitant DAPK up-regulation. When DAPK was over-expressed, A549/CDDP cells became sensitive to cisplatin and the cytotoxicity of TSA could be increased. Moreover, the cytotoxicity of TSA could be alleviated by inhibition of DAPK activity by the expression of a recombinant C-terminal fragment of DAPK or RNA interference. CONCLUSION: TSA induced sensitivity to cisplatin treatment in cisplatin-resistant A549 cells. The up-regulation of DAPK is one of the mechanisms mediating sensitization to TSA-induced apoptosis in cisplatin-resistant cells.
Assuntos
Antineoplásicos/farmacologia , Cisplatino/farmacologia , Ácidos Hidroxâmicos/farmacologia , Inibidores da Síntese de Proteínas/farmacologia , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/fisiopatologia , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/genética , Proteínas Quinases Dependentes de Cálcio-Calmodulina/efeitos dos fármacos , Proteínas Quinases Dependentes de Cálcio-Calmodulina/genética , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Proteínas Quinases Associadas com Morte Celular , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/fisiopatologia , Regulação para Cima/efeitos dos fármacosRESUMO
T cell DNA methylation levels decline with age, activating genes such as KIR and TNFSF7 (CD70), implicated in lupus-like autoimmunity and acute coronary syndromes. The mechanisms causing age-dependent DNA demethylation are unclear. Maintenance of DNA methylation depends on DNA methyltransferase 1 (Dnmt1) and intracellular S-adenosylmethionine (SAM) levels, and is inhibited by S-adenosylhomocysteine (SAH). SAM levels depend on dietary micronutrients including folate and methionine. SAH levels depend on serum homocysteine concentrations. T cell Dnmt1 levels also decline with age. We hypothesized that age-dependent Dnmt1 decreases synergize with low folate, low methionine or high homocysteine levels to demethylate and activate methylation-sensitive genes. T cells from healthy adults ages 22-81, stimulated and cultured with low folate, low methionine, or high homocysteine concentrations showed demethylation and overexpression of KIR and CD70 beginning at age approximately 50 and increased further with age. The effects were reproduced by Dnmt1 knockdowns in T cells from young subjects. These results indicate that maintenance of T cell DNA methylation patterns is more sensitive to low folate and methionine levels in older than younger individuals, due to low Dnmt1 levels, and that homocysteine further increases aberrant gene expression. Thus, attention to proper nutrition may be particularly important in the elderly.
Assuntos
Síndrome Coronariana Aguda/genética , Envelhecimento/metabolismo , Doenças Autoimunes/genética , Metilases de Modificação do DNA/metabolismo , Regulação da Expressão Gênica/fisiologia , Micronutrientes/metabolismo , Síndrome Coronariana Aguda/fisiopatologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Doenças Autoimunes/fisiopatologia , Ligante CD27/metabolismo , Antígenos CD28/metabolismo , Células Cultivadas , DNA (Citosina-5-)-Metiltransferase 1 , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA , Feminino , Ácido Fólico/metabolismo , Homocisteína/metabolismo , Humanos , Masculino , Metionina/metabolismo , Pessoa de Meia-Idade , Receptores KIR/genética , Linfócitos T/citologia , Linfócitos T/metabolismoRESUMO
OBJECTIVE: To establish a rapid polymerase chain reaction (PCR)method for detection of staphylococcus aureus in milk, ice cream and meat. METHODS: Two pairs of oligonucleotide primers were designed with thermo nuclease gene nuc and surfaced-associated fibrinogen-binding protein gene ClfA to detect staphylococcus aureus. Fifty-two staphylococcus aureus and thirty-one non staphylococcus aureus were amplified by PCR to verify the specificity. Various numbers of bacteria were added into milk, ice cream and meat. After enrichment, DNA extracted in different time was amplified by PCR to verify detection limit. RESULTS: Each primer pair allows specific detection. The limit of detection was 10 cfu/g (ml) in three kinds of food. Whole procedure of detection could be finished in 24 hours. CONCLUSION: A rapid, sensitive and specific PCR method can be applied detecting staphylococcus aureus in milk, ice cream and meat.
Assuntos
Contaminação de Alimentos/análise , Microbiologia de Alimentos , Reação em Cadeia da Polimerase , Staphylococcus aureus/isolamento & purificação , Animais , Sorvetes/microbiologia , Carne/microbiologia , Leite/microbiologia , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Recent studies have shown that NS-398, a highly selective COX-2 inhibitor, can enhance the inhibition effects of cisplatin on adenocarcinoma cell proliferation, but the mechanism is still unknown. The aim of this study is to explore the mechanism about the synergistic effects of NS-398 and cisplatin to human lung adenocarcinoma cell line. METHODS: Differentially expressed proteins were separated in human lung adenocarcinoma cells treated with NS-398 and/or cisplatin by immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE), then 19 out of obtained proteins were identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: There were 22 down-regulated proteins and 2 up-regulated proteins in NS-398+cisplatin combination group compared with control groups. Finally six proteins were identified, in which some were cytokeratins, some were associated with proliferation and apoptosis, and the others were involved in metabolism of tumor cells. CONCLUSIONS: The down-regulation of HSP90 and triosephosphate isomerase may be related to the synergistic effects of NS-398 and cisplatin on human lung adenocarcinoma cells.