RESUMO
Bacterial genome data are accumulating at an unprecedented speed due to the routine use of sequencing in clinical diagnoses, public health surveillance, and population genetics studies. Genealogical reconstruction is fundamental to many of these uses; however, inferring genealogy from large-scale genome data sets quickly, accurately, and flexibly is still a challenge. Here, we extend an alignment- and annotation-free method, PopPUNK, to increase its flexibility and interpretability across data sets. Our method, iterative-PopPUNK, rapidly produces multiple consistent cluster assignments across a range of sequence identities. By constructing a partially resolved genealogical tree with respect to these clusters, users can select a resolution most appropriate for their needs. We showed the accuracy of clusters at all levels of similarity and genealogical inference of iterative-PopPUNK based on simulated data and obtained phylogenetically concordant results in real data sets from seven bacterial species. Using two example sets of Escherichia/Shigella and Vibrio parahaemolyticus genomes, we show that iterative-PopPUNK can achieve cluster resolutions ranging from phylogroup down to sequence typing (ST). The iterative-PopPUNK algorithm is implemented in the "PopPUNK_iterate" program, available as part of the PopPUNK package.
Assuntos
Algoritmos , Genoma Bacteriano , Bactérias/genética , Análise por ConglomeradosRESUMO
BACKGROUND: Maternal parenting self-efficacy plays a critical role in facilitating positive parenting practices and successful adaption to motherhood. The Perceived Maternal Parenting Self-Efficacy Scale (PMPS-E), as a task-specific measure, confirms its psychometric properties in cultural contexts. Compared with other tools, the advantages of the PMPS-E are as follows: (i) specific context or time period during the lifespan of a child, (ii) explicitly assess parenting self-efficacy across a diverse enough range of parenting tasks or activities during the perinatal/postnatal period and (iii) having robust psychometric properties. The aim of this study was to translate and determine the psychometric properties of the PMPS-E among Chinese postpartum women (C-PMPS-E). METHOD: The cross-cultural adaptation process followed Beaton et al.'s intercultural debugging guidelines. A total of 471 women were included to establish the psychometric properties of the C-PMPS-E. Mothers were asked to complete the C-PMPS-E, Edinburgh Postnatal Depression Scale (EPDS), the Generalized Anxiety Disorder-7 (GAD-7) and several demographic questions. The psychometric testing of the C-PMPS-E was established through item analysis, construct validity and internal consistency reliability. RESULTS: Item analysis showed that the critical ratios of all items were greater than 3 between the low-score group and high-score group, and all item-total correlation coefficients were greater than 0.4. The fit indices showed that the original correlated four-factor model of C-PMPS-E was observed to be an excellent fit to the data. The PMPS-E was negatively correlated with the EPDS and GAD-7 demonstrating its discriminant validity. As expected, no significant correlation was found between PMPS-E total or subscale scores and mothers' age. In addition, statistically significant differences for parity were detected for C-PMPS-E total and subscale scores with multipara having higher scores. This was taken as further evidence of the scale known-groups discriminant validity. In terms of internal consistency, the Cronbach's alpha of the C-PMPS-E total scale was 0.950, and subscales ranged from 0.76 to 0.89. Furthermore, a ROC curve analysis was conducted to establish the ability of the C-PMPS-E to distinguish between symptoms of depression and symptoms of anxiety. A cut-off value of 55 was identified that resulted in good specificity and fair sensitivity. CONCLUSION: The C-PMPS-E is a reliable and valid tool to assess maternal parenting self-efficacy in a Chinese context.
Assuntos
Mães , Poder Familiar , Período Pós-Parto , Psicometria , Autoeficácia , Humanos , Feminino , Adulto , Poder Familiar/psicologia , Período Pós-Parto/psicologia , Reprodutibilidade dos Testes , Mães/psicologia , China , Inquéritos e Questionários/normas , Adulto Jovem , Traduções , Depressão Pós-Parto/psicologia , Depressão Pós-Parto/diagnósticoRESUMO
The enterohemorrhagic Escherichia coli pathotype is responsible for severe and dangerous infections in humans. Establishment of the infection requires colonization of the gastro-intestinal tract, which is dependent on the Type III Secretion System. The Type III Secretion System (T3SS) allows attachment of the pathogen to the mammalian host cell and cytoskeletal rearrangements within the host cell. Blocking the functionality of the T3SS is likely to reduce colonization and therefore limit the disease. This route offers an alternative to antibiotics, and problems with the development of antibiotics resistance. Salicylidene acylhydrazides have been shown to have an inhibitory effect on the T3SS in several pathogens. However, the main target of these compounds is still unclear. Past work has identified a number of putative protein targets of these compounds, one of which being WrbA. Whilst WrbA is considered an off-target interaction, this study presents the effect of the salicylidne acylhydrazide compounds on the activity of WrbA, along with crystal structures of WrbA from Yersinia pseudotuberculosis and Salmonella serovar Typhimurium; the latter also containing parts of the compound in the structure. We also present data showing that the original compounds were unstable in acidic conditions, and that later compounds showed improved stability.
Assuntos
Escherichia coli Êntero-Hemorrágica , Proteínas de Escherichia coli , Yersinia pseudotuberculosis , Animais , Antibacterianos/metabolismo , Escherichia coli Êntero-Hemorrágica/metabolismo , Proteínas de Escherichia coli/metabolismo , Humanos , Mamíferos/metabolismo , Proteínas Repressoras/metabolismo , Salmonella typhimurium/metabolismo , Sistemas de Secreção Tipo III/metabolismo , Yersinia pseudotuberculosis/metabolismoRESUMO
We propose and experimentally demonstrate that giant asymmetric reflection of circularly polarized light based on asymmetric coupling can be achieved in single-layer extrinsic chiral metasurfaces at oblique incidence. The asymmetric coupling and asymmetric reflection in the extrinsic chiral metasurfaces are caused by extrinsic chirality, allowing them to have extremely high values. An asymmetric reflection of approximately 40% is measured. Furthermore, the asymmetric reflection of extrinsic chiral metasurfaces is demonstrated not only in intensity but also in phase retardation, which induces asymmetric polarization state conversion. An approximately 14° asymmetric reflected polarization offset from the symmetry axis is achieved. Our research provides an effective new method for constructing huge asymmetric coupled systems to manipulate electromagnetic waves.
RESUMO
BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is a lethal cancer. Approximately 80% of patients initially diagnosed with locally advanced or metastatic disease survive only 4-11 months after diagnosis. Tremendous efforts have been made toward understanding the biology of PDAC. RESULTS: In this study, we first utilized next-generation sequencing technique and existing microarray datasets to identify significant differentially expressed genes between PDAC and non-tumor adjacent tissue. By comparing top significant survival genes in PDAC Gene Expression Profiling Interactive Analysis database and PDAC transcriptome data from patients, our integrated analysis discovered five potential central genes (i.e., MYEOV, KCNN4, FAM83A, S100A16, and DDX60L). Subsequently, we analyzed the cellular functions of the potential novel oncogenes MYEOV and DDX60L, which are highly expressed in PDAC cells. Notably, the knockdown of MYEOV and DDX60L significantly inhibited the metastasis of cancer cells and induced apoptosis. Further RNA sequencing analyses showed that massive signaling pathways, particularly the TNF signaling pathway and nuclear factor-kappa B (NF-κB) signaling pathway, were affected in siRNA-treated cancer cells. The siDDX60L and siMYEOV significantly inhibited the expression of chemokine CXCL2, which may potentially affect the tumor microenvironment in PDAC tissues. CONCLUSIONS: The present findings identified the novel oncogene DDX60L, which was highly expressed in PDAC. Transcriptome profiling through siRNA knockdown of DDX60L uncovered its functional roles in the PDAC in humans.
Assuntos
Carcinoma Ductal Pancreático , Oncogenes , Neoplasias Pancreáticas , Biomarcadores Tumorais , Carcinoma Ductal Pancreático/genética , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Humanos , Proteínas de Neoplasias , Neoplasias Pancreáticas/genética , Microambiente TumoralRESUMO
BACKGROUND AND AIMS: Hepatocellular carcinoma (HCC) is linked to immunosuppression. Relieving immunosuppression has been an attractive strategy to improve the efficacy of cancer immunotherapy. Peptidoglycan recognition protein 2 (PGLYRP2) is a pattern recognition receptor which is specifically expressed in liver and implicated in the regulation of innate immunity and immunosurveillance. However, the role of hepatic PGLYRP2 in modulating immune responses against HCC remains to be investigated. APPROACH AND RESULTS: In this study, we investigated whether PGLYRP2 is able to influence HCC progression through regulating host antitumor immune responses. We demonstrated that PGLYRP2 was down-regulated in HCC, which was linked with poor prognosis in patients (P < 0.001). PGLYRP2 overexpression in HCC cells significantly enhanced antitumor immune responses in immune-competent mice and elevated immune response rates of peripheral blood mononuclear cells against HCC. Mechanistically, DNA methyltransferase 3A-mediated promoter hypermethylation was responsible for the down-regulation of PGLYRP2 in HCC. PGLYRP2 promoted production of chemokine (C-C motif) ligand 5 (CCL5) in HCC through binding to the CCL5 promoter, which contributed to the enhanced antitumor immunity. CONCLUSIONS: We provide evidence that tumor-derived PGLYRP2 acts as a candidate biomarker for adequate immune response against HCC and improved patient outcomes, indicating the importance of hepatic PGLYRP2 in cancer immunosurveillance and in designing immunotherapeutic approaches.
Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/imunologia , Proteínas de Transporte/metabolismo , Vigilância Imunológica , Neoplasias Hepáticas/imunologia , Proteínas Supressoras de Tumor/metabolismo , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/terapia , Proteínas de Transporte/genética , Linhagem Celular Tumoral , Quimiocina CCL5/genética , Metilação de DNA , DNA Metiltransferase 3A , Regulação Neoplásica da Expressão Gênica , Humanos , Imunoterapia , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/terapia , Ativação Linfocitária , Prognóstico , Regiões Promotoras Genéticas , Linfócitos T/imunologia , Proteínas Supressoras de Tumor/genéticaRESUMO
BACKGROUND: The growth- and plasticity-associated protein-43 (GAP43) is biasedly expressed in indigestive system and nervous system. Recent study has shown that GAP43 is responsible for the development of neuronal growth and axonal regeneration in normal nervous tissue, while serves as a specific biomarker of relapsed or refractory neuroblastoma. However, its expression pattern and function in digestive system cancer remains to be clarified. METHODS: In this study, we examined the GAP43 status with qRT-PCR and bisulfite genomic sequencing in colorectal cancer (CRC). We investigated the effect of overexpressed GAP43 in CRC cells with RNA-seq. The RNA-seq data was analyzed with DAVID and IPA. RESULTS: GAP43 was downregulated in CRC compared to the adjacent tissues. DNA methylase inhibitor 5-Aza-CdR treatment could significantly induce GAP43, indicated that the silencing of GAP43 gene in CRC is closely related to DNA methylation. Bisulfite genomic sequencing confirmed the promoter methylation of GAP43 in CRC. To explore the transcriptional alterations by overexpressed GAP43 in CRC, we performed RNA-seq and found that upregulated genes were significantly enriched in the signaling pathways of ABC transporters and ECM-receptor interaction, while downregulated genes were significantly enriched in Ribosome signaling pathway. Further Ingenuity Pathway Analysis (IPA) showed that EIF2 signaling pathway was significantly repressed by overexpression of GAP43. CONCLUSION: Our findings provide a novel mechanistic insight of GAP43 in CRC. Transcriptome profiling of overexpressed GAP43 in CRC uncovered the functional roles of GAP43 in the development of human CRC.
Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/patologia , Metilação de DNA , Fator de Iniciação 2 em Eucariotos/metabolismo , Proteína GAP-43/metabolismo , Regulação Neoplásica da Expressão Gênica , Transportadores de Cassetes de Ligação de ATP/genética , Apoptose , Biomarcadores Tumorais/genética , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Fator de Iniciação 2 em Eucariotos/genética , Proteína GAP-43/genética , Redes Reguladoras de Genes , Humanos , Prognóstico , Regiões Promotoras Genéticas , Transcriptoma , Células Tumorais CultivadasRESUMO
Cisplatin is a widely used chemotherapeutic drug in lung cancer treatment. Most cancer patients eventually develop cisplatin resistance, resulting in a poor prognosis. Previously, we identified a novel marker, family with sequence similarity 60A (FAM60A), that was responsible for resistance in cisplatin-resistant human lung adenocarcinoma A549 (A549/DDP) cells. Here, we investigated the biological effects of FAM60A in A549/DDP cells and explored the underlying molecular mechanisms to understand its functional role in cisplatin resistance. Real-time quantitative PCR and western blot analysis were used to determine the expression levels of FAM60A in A549/DDP cells. FAM60A and SKP2 were knockdown with small-interfering RNA (siRNA). Cancer cell viability was analyzed with flow cytometry. The mRNA and protein expression levels of FAM60A increased significantly and dose-dependently in A549/DDP cells following cisplatin treatment. FAM60A overexpression up-regulated MDR1 expression, inhibited caspase 3, cleaved-caspase 3, and caspase 8 expression, and prevented cancer cell death. Microarray analysis of cells transfected with siRNA against the FAM60A transcript and control samples showed that SKP2 expression was positively regulated by FAM60A. SKP2 knockdown using a short-hairpin RNA reversed the functions induced by FAM60A. These results suggest that overexpression of FAM60A in A549/DDP cells led to SKP2 upregulation and enhanced cisplatin resistance in cancer cells. These provide new insights into chemoresistance and may contribute to reversing cisplatin resistance during lung cancer treatment.
Assuntos
Antineoplásicos/farmacologia , Cisplatino/farmacologia , Proteínas de Ligação a DNA/metabolismo , Resistencia a Medicamentos Antineoplásicos , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/tratamento farmacológico , Proteínas Quinases Associadas a Fase S/metabolismo , Apoptose , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Proliferação de Células , Proteínas de Ligação a DNA/genética , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Proteínas Quinases Associadas a Fase S/genética , Células Tumorais CultivadasRESUMO
Background: Fractional carbon dioxide laser resurfacing (FxCR) is a routine treatment of Dermatology while many patients suffered the damage of skin barrier function after FxCR. Objective: To evaluate the benefits of antimicrobial peptides (AMPs) and hyaluronic acid (HA) compound mask on wound healing after FxCR on human and mouse skin. Methods: Twenty-four subjects were treated with FxCR on the bilateral cheeks. AMPs and HA compound mask was applied on the FxCR-treated area of left cheek. The erythema index (EI), melanin index (MI), transepidermal water loss (TEWL) of FxCR-treated areas on both cheeks were measured. By HE staining, immunohistostaing and western blotting, we analyzed epidermal thickness, FLG, IVL expression and protein levels of cramp in FxCR treated dorsal mice skin. Results: The EI, MI, and TEWL in the AMPs and HA compound mask-treated area of left cheek were significantly lower than those in the untreated area of right cheek. Topically application of AMPs and HA compound mask reduced thickening of mouse skin and also result in an increase in the production of FLG, IVL and cramp. Conclusion: Application of AMPs and HA compound mask is an effective method for enhancing wound healing after FxCR, by reducing transient adverse effects such as erythema, hyperpigmentation, and increased TEWL.
Assuntos
Anti-Infecciosos/uso terapêutico , Bochecha , Ácido Hialurônico/uso terapêutico , Lasers de Gás/uso terapêutico , Regeneração da Pele por Plasma/métodos , Cicatrização/efeitos dos fármacos , Adulto , Animais , Dióxido de Carbono , Eritema/etiologia , Feminino , Proteínas Filagrinas , Humanos , Proteínas de Filamentos Intermediários/metabolismo , Masculino , Melaninas/metabolismo , Camundongos , Perda Insensível de ÁguaRESUMO
Curcumin is a potent anti-cancer drug in several types of human cancers. Despite of several preclinical and clinical studies of curcumin, the precise mechanism of curcumin in cancer prevention has remained unclear. In our study, we for the first time investigated whole transcriptome alteration in A549 non-small cell lung cancer (NSCLC) cell lines after treatment with curcumin using RNA sequencing. We found that lots of genes and signaling pathways were significantly altered after curcumin treatment in A549 cells. With bioinformatics approaches (gene ontology, Kyoto Encyclopedia of Genes and Genomes, and STRING), we found that those curcumin altered genes were not only the genes that induce cell death but also those extracellular matrix receptors and mitogen-activated protein kinase signaling pathway genes which regulate cell migration and proliferation. Among those significantly altered genes, eight genes ( COL1A1, COL4A1, COL5A1, LAMA5, ITGA3, ITGA2B, DDIT3, and DUSP1) were further examined by quantitative reverse transcription polymerase chain reaction and western blot analysis in four non-small cell lung cancer cell lines. Both in cell lines and in mouse model, the extracellular matrix receptors including the integrin ( ITGA3 and ITGA2B), collagen ( COL5A1), and laminin ( LAMA5) were significantly inhibited by curcumin at messenger RNA and protein levels. Functional studies confirmed that curcumin not only induced A549 cell death but also repressed cell proliferation and migration by regulating extracellular matrix receptors. Collectively, our study suggests that curcumin may be used as a promising drug candidate for intervening lung cancer in future studies.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Colágeno Tipo V/biossíntese , Curcumina/administração & dosagem , Integrina alfa2/biossíntese , Integrina alfa3/biossíntese , Laminina/biossíntese , Células A549 , Animais , Apoptose/efeitos dos fármacos , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Colágeno Tipo V/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Integrina alfa2/genética , Integrina alfa3/genética , Laminina/genética , Camundongos , RNA Mensageiro/biossíntese , Receptores de Superfície Celular/antagonistas & inibidores , Receptores de Superfície Celular/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Glucocorticoid-induced osteoporosis is a common form of secondary osteoporosis. Glucocorticoids affect both bone formation and resorption, and prolonged glucocorticoid exposure can suppress osteoblast activities. beta-Ecdysone, found in many plants, is involved in protein synthesis, carbohydrate and lipid metabolism, and immunologic modulation. Here, we evaluated the effects of beta-ecdysone on osteoblast viability by assessing apoptosis following treatment with excess glucocorticoids. Mouse bone marrow stromal cells were induced to differentiate and grow into osteoblasts, and then treated with 10 µM glucocorticoid and 10, 1, or 0.1 µM beta-ecdysone. The expression levels of osteoblast growth and differentiation factors (runt-related transcription factor 2, osteogenic protein-1, and alkaline phosphatase), apoptosis-related genes (transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8), and Akt1 and phospho-Akt (Thr308) were then assessed via alkaline phosphatase staining, acridine orange-propidium iodide staining, annexin V/PI apoptosis assay, real-time RT-PCR, and Western blot analyses. Notably, treatment with 10 µM glucocorticoid resulted in reduced osteoblast viability and the specific activity of alkaline phosphatase as well as reduced runt-related transcription factor 2, osteogenic protein-1, and alkaline phosphatase mRNA expression in vitro, indicating that glucocorticoid inhibited osteogenic differentiation. Moreover, glucocorticoid treatment yielded increased transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8 expression and decreased Akt1 and phospho-Akt levels, indicating glucocorticoid-induced apoptosis. Meanwhile, beta-ecdysone inhibited glucocorticoid function, preserving the expression of Akt1 and phospho-Akt and reducing the expression of transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8. Thus, beta-ecdysone prevented glucocorticoid-induced osteoblast apoptosis in vitro. These data highlight the potential for beta-ecdysone as a treatment for preventing the effects of glucocorticoid on bone growth.
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Apoptose/efeitos dos fármacos , Ecdisterona/farmacologia , Glucocorticoides/antagonistas & inibidores , Osteoblastos/efeitos dos fármacos , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Glucocorticoides/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mifepristona/farmacologia , Plantas Medicinais/químicaRESUMO
BACKGROUND/AIMS: Myocardial infarction (MI) is a serious complication of atherosclerosis associated with increasing mortality attributable to heart failure. This study is aimed to assess the global changes in and characteristics of the transcriptome of circular RNAs (circRNAs) in heart tissue during MI induced heart failure (HF). METHODS: Using a post-myocardial infarction (MI) model of HF in mice, we applied microarray assay to examine the transcriptome of circRNAs deregulated in the heart during HF. We confirmed the changes in circRNAs by quantitative PCR. RESULTS: We revealed and confirmed a number of circRNAs that were deregulated during HF, which suggests a potential role of circRNAs in HF. CONCLUSIONS: The distinct expression patterns of circulatory circRNAs during HF indicate that circRNAs may actively respond to stress and thus serve as biomarkers of HF diagnosis and treatment.
Assuntos
Perfilação da Expressão Gênica/métodos , Insuficiência Cardíaca/genética , Infarto do Miocárdio/complicações , Miocárdio/metabolismo , RNA/genética , Animais , Análise por Conglomerados , Insuficiência Cardíaca/etiologia , Masculino , Camundongos Endogâmicos C57BL , MicroRNAs/genética , Miocárdio/patologia , Análise de Sequência com Séries de Oligonucleotídeos/métodos , RNA/classificação , RNA Circular , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND/AIMS: Ginsenoside Rb1 (GS-Rb1) is one of the most important active pharmacological extracts of the Traditional Chinese Medicine ginseng, with extensive evidence of its cardioprotective properties. Mir-208 has been shown to act as a biomarker of acute myocardial infarction in vivo studies including man. However the impact of miR-208 on the protective effect of GS-Rb1 in hypoxia/ischemia injured cardiomyocytes remains unclear. The current study aims to investigate the target gene of miR-208 and the impact on the protective effect of GS-Rb1 in hypoxia/ischemia (H/I) injuried cardiomyocytes. MATERIALS AND METHODS: Primary cultures of neonatal rat cardiomyocytes (NRCMs) was subjected to the H/I conditions with or without GS-Rb1. Cell viability was calculated by MTT assay and confirmed by flow cytometry analysis. Mir-208 was then detected by qRT-PCR. Luciferase reporter assay was carried out to detect the target gene of Mir-208. Then the NRCMs were transfected with miR-208 mimics and inhibitors to evaluate the impact on cardioprotective properties of Rb1. RESULTS: The miR-208 expression level was clearly upregulated in the H/I treated NRCMs accompanied by the percentage of the apoptotic cells which could be reversed by GS-Rb1 pretreatment. The nemo-like kinase (NLK) mRNA and protein expression levels were decreased in H/I group measured by RT-PCR and western blotting. Luciferase activity assay was then carried out to identify that NLK may be a direct target of mir-208. MTT assay showed that miR-208 inhibitor slightly decreased the protective effect of Rb1 on the H/I impaired NRCMs. However, results showed no statistical difference. CONCLUSIONS: These findings proved that NLK was a direct target of mir-208 and miR-208 act indirectly during Rb1 protecting H/I impaired NRCMs and further researches were needed to explore the relationship that microRNAs and other signal pathways in the protective effect of GS-Rb1 on the hypoxia/ischemia injuries in cardiomyocytes.
Assuntos
Cardiotônicos/farmacologia , Ginsenosídeos/farmacologia , MicroRNAs/genética , Miócitos Cardíacos/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/genética , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Apoptose/genética , Sequência de Bases , Sítios de Ligação , Hipóxia Celular , Regulação da Expressão Gênica , Genes Reporter , Luciferases/genética , Luciferases/metabolismo , MicroRNAs/antagonistas & inibidores , MicroRNAs/metabolismo , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Antissenso/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Ratos , Ratos Sprague-Dawley , Transdução de SinaisRESUMO
Circulating tumor DNA (ctDNA) is becoming an important biomarker in noninvasive diagnosis and monitoring of tumor dynamics. This study tested the feasibility of plasma ctDNA for the non-invasive analysis of tumor mutations in esophageal squamous cell carcinoma (ESCC) by sequencing of tumor, tumor-adjacent, and normal tissue, as well as pre-surgery and post-surgery plasma. Exome sequencing of eight patients identified between 29 and 134 somatic mutations in ESCCs, many of which were also determined in ctDNA. Comparison of pre-surgery and post-surgery plasma has shown that mutations had reduced frequency or disappeared after surgery treatment. We further evaluated the TruSight Cancer sequencing panel by using it to detect mutations in the plasma of three patients. Tumor mutations were only found in one of them. To design a sequencing panel with improved targeting, we identified significantly mutated genes by meta-analysis of 532 ESCC genomes. Our results confirmed the well-known driver genes and found several uncharacterized genes. The new panel consisted of 90 recurrent genes, which theoretically achieved 94% and 75% of sensitivity when detecting at least 1 and 2 mutant genes in ESCC patients, respectively. Our results demonstrate the feasibility of using ctDNA to detect ESCCs and monitor treatment effect. The low-cost and sensitive target panel could facilitate clinical usage of ctDNA as a noninvasive biomarker.
Assuntos
Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/diagnóstico , DNA de Neoplasias/genética , Neoplasias Esofágicas/diagnóstico , Biomarcadores Tumorais/sangue , Carcinogênese/genética , Carcinoma de Células Escamosas/sangue , Carcinoma de Células Escamosas/genética , Análise Mutacional de DNA/métodos , DNA de Neoplasias/sangue , Neoplasias Esofágicas/sangue , Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas do Esôfago , Genes Neoplásicos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mutação , Recidiva Local de Neoplasia/diagnóstico , Recidiva Local de Neoplasia/genéticaRESUMO
BACKGROUND/AIMS: Licorice has been used to treat many diseases, including palpitations, in both Eastern and Western societies for thousands of years. It has been reported that glycyrrhetinic acid (GA), an aglycone saponin extracted from licorice root, exerts protective effects on the cardiovascular system, limits infarct sizes and protects against the development of arrhythmia. However, the mechanisms underlying the effects of glycyrrhetinic acid on the cardiovascular system remain poorly understood. This study aimed to determine the mechanisms underlying the protective effects of GA against lethal cardiac arrhythmias induced via ischemia-reperfusion in rat hearts, and to examine its electropharmacological properties. MATERIALS AND METHODS: Anesthetized rats were divided into control (CTL), GA5, GA10, and GA20 groups. GA was administered intravenously 15 min before the occlusion of the left anterior descending coronary artery, at dosages of 5, 10 and 20 mg/kg, respectively. Single ventricular myocytes were isolated using enzymolysis. The whole-cell patch clamp technique was utilized to record Ica, L, Ito and action potentials (APs). RESULTS: During reperfusion, the incidence of ventricular fibrillation (VF) was decreased in each of the groups compared with the CTL group (p<0.05). The ventricular tachycardia (VT)/VF score was significantly decreased in the GA20 group. Action potential durations (APDs) were prolonged by GA; both L-type calcium current (Ica-L) and transient outward potassium current (Ito) were blocked in a concentration-dependent manner by GA. CONCLUSION: These results suggest that GA attenuates both the susceptibility to and the incidence of fatal ventricular arrhythmia during reperfusion in rat hearts via the prolongation of the APD and the inhibition of both Ica-L and Ito. GA appears to be a promising antiarrhythmic agent in the setting of ischemia/reperfusion.
Assuntos
Antiarrítmicos/uso terapêutico , Arritmias Cardíacas/etiologia , Arritmias Cardíacas/prevenção & controle , Ácido Glicirretínico/uso terapêutico , Células Musculares/efeitos dos fármacos , Traumatismo por Reperfusão Miocárdica/complicações , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Potenciais de Ação/efeitos dos fármacos , Animais , Arritmias Cardíacas/metabolismo , Arritmias Cardíacas/patologia , Cálcio/metabolismo , Canais de Cálcio Tipo L/metabolismo , Células Cultivadas , Incidência , Células Musculares/metabolismo , Células Musculares/patologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Ratos Sprague-Dawley , Taquicardia Ventricular/etiologia , Taquicardia Ventricular/metabolismo , Taquicardia Ventricular/patologia , Taquicardia Ventricular/prevenção & controle , Fibrilação Ventricular/etiologia , Fibrilação Ventricular/metabolismo , Fibrilação Ventricular/patologia , Fibrilação Ventricular/prevenção & controleRESUMO
The study aimed to clarify the relationship between ß-elemene, a long noncoding RNA (lncRNA), and human telomerase reverse transcriptase (hTERT) in esophageal carcinoma ECA-109 cells. The proliferation of ECA-109 cells was measured using a CCK-8 kit and flow cytometry. PCR microarray and real-time RT-PCR were designed to determine lncRNA expression in ECA-109 cells before and after treatment with ß-elemene. Western blot was used to detect the hTERT level after the differentially expressed lncRNAs in ECA-109 cells were interfered with small interfering RNA (siRNA). On treatment with ß-elemene, the proliferation of ECA-109 cells was notably inhibited, and about 85% of the lncRNAs showed higher expression levels in ECA-109 cells than in those untreated cells, from which, CDKN2B-AS1 was screened out. A specific siRNA (si-CDKN2B-AS1) that targets the ß-elemene-mediated lncRNA CDKN2B-AS1 was designed, synthesized, and applied to treat ECA-109 cells. Its interference efficiency reached as high as 89.6%. When ECA-109 cells were transfected with the siRNA, the hTERT level was increased by 84.7%. The CCK-8 assay showed that the proliferation of ECA-109 cells treated with ß-elemene was significantly promoted after siRNA transfection (P<0.01). It was also shown by flow cytometry that, compared with the scramble-treated group (negative control), the proliferation index value of ECA-109 cells in the si-CDKN2B-AS1 treatment group was notably increased (25.7 vs. 51.7%) and the TERT protein level was increased by 67.25% after the cells were treated with si-CDKN2B-AS1. The chemotherapeutic drug ß-elemene suppressed the proliferation of esophageal carcinoma ECA-109 cells by regulating the inhibition of hTERT expression by lncRNA CDKN2B-AS1.
Assuntos
Antineoplásicos/farmacologia , RNA Longo não Codificante/metabolismo , Sesquiterpenos/farmacologia , Telomerase/metabolismo , Carcinoma de Células Escamosas , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias Esofágicas , Regulação da Expressão Gênica , Humanos , RNA Longo não Codificante/genética , Telomerase/genéticaRESUMO
BACKGROUND: Chronic bronchitis is a type of common chronic inflammatory respiratory disease, which is mainly characterized by chronic cough and expectoration. Clinical practice and experimental research have shown that the modified tonifying spleen-lung method has significant preventive and therapeutic effects on chronic lung diseases, but the mechanism of TSLR in the treatment of chronic bronchitis are not yet clear. OBJECTIVE: To explore the mechanism of tonifying spleen-lung recipe (TSLR) in the treatment of chronic bronchitis (CB) through network pharmacology combined with observational studies. MATERIALS AND METHODS: The effective components, core targets and signaling pathways of TSLR in the treatment of chronic bronchitis were obtained using network pharmacology. One hundred and thirty-seven elderly CB patients were selected as the observational group who were treated by TSLR, and 67 no-CB cases from the Physical Examination Center were selected as the control group. We compared the levels of inflammatory parameters between patients before and after TSLR treatment, and after treatment group with the control group were also compared. RESULTS: The key effective components of TSLR selected by network pharmacology included quercetin, kaempferol, luteolin, and nobiletin, and the core targets involved HSP90AA1, AKT1, JUN, MAPK1, IL6, MAPK3, MAPK14, STAT1, NFKB1, and CDKN1. KEGG pathway enrichment analysis revealed that the TNF signaling pathway, PI3K-AKT and AGE-RAGE signaling pathways might play a key role in the treatment of CB. The observation study demonstrated that compared with the control group, the levels of WBC, NEU, NLR, PCT, and CRP in the research group after TSLR treatment were increased. Although the levels of WBC, NEU, NLR, and PCT in the research group after TSLR treatment were higher than those in the control group, the above indicators trend tended towards the control group, and there was no significant difference in CRP indicators between the control group and after treatment group. CONCLUSION: TSLR had a good therapeutic effect on chronic bronchitis patients, which might be related to the fact that the natural active components in TSLR inhibit inflammation by regulating the expression of proteins related to PI3K-AKT and TNF signaling pathways.
RESUMO
OBJECTIVE: To explore the effects of electric stimulation and biofeedback therapy in patients with postpartum pelvic organ prolapse and to identify factors that can affect therapeutic efficacy outcomes. METHOD: This retrospective study analysed clinical data about patients with postpartum pelvic organ prolapse. A total of 328 women with pelvic organ prolapse at 6 weeks postpartum were recruited from one tertiary hospitals in Sichuan province in China, between March 2019 and March 2022. The prognostic factors of therapeutic efficacy were analysed using logistic regression and decision tree model. RESULTS: Overall, 259 women showed clinical benefits from the treatment. The logistic regression model showed that parity, pelvic floor muscle training at home, and the pelvic organ prolapse quantitation stage before treatment were independent prognostic factors. The decision tree model showed that the pelvic organ prolapse quantitation stage before treatment was the main prognostic factor, followed by parity. There was no significant difference in the area under the receiver operating characteristic curve between the two models. CONCLUSION: Parity, pelvic floor muscle training at home, and the pelvic organ prolapse quantitation stage before treatment were important prognostic factors of electric stimulation and biofeedback therapy on postpartum pelvic organ prolapse.
Assuntos
Biorretroalimentação Psicológica , Terapia por Estimulação Elétrica , Prolapso de Órgão Pélvico , Humanos , Feminino , Prolapso de Órgão Pélvico/terapia , Estudos Retrospectivos , Adulto , Prognóstico , Biorretroalimentação Psicológica/métodos , Terapia por Estimulação Elétrica/métodos , Resultado do Tratamento , Diafragma da Pelve/fisiopatologia , Período Pós-Parto , China , Paridade , Modelos Logísticos , GravidezRESUMO
Reducing the south and reinforcing the north method (RSRN) has a positive effect on atherosclerosis disease. However, there is a lack of objective standards based on the quantification of 4 diagnostic methods in evaluating the improvement or effectiveness of the treatment. This study aimed to explore the quantitative evaluation of the therapeutic effect of RSRN on postmenopausal atherosclerosis based on the 4 diagnostic methods. The observational prospective cohort study was conducted at Longhua hospital Shanghai University of traditional Chinese medicine. According to the inclusion criteria, 96 patients (disease group) and 38 healthy cases (control group) were selected, the pulse parameters were compared between the 2 groups to demonstrate the reliability and success of the disease model. Then 4 diagnostic information before and after RSRN treatment were collected and statistical analyzed by 1-way analysis of variance (ANOVA) (with Bonferroni correction). Furthermore, social network analysis was used to analyze the changes of symptoms, tongue, pulse, and complexion characteristics before and after treatment. There was a significant difference in pulse parameters between the disease group and the control group. The pulse parameters t1, h3, h3/h1, h4/h1, S, As, and w values in disease group were higher than those in control group, while the h5, h5/h1, and Ad values were lower than those in control group (Pâ <â .05). After the treatment of RSRN, the clinical symptoms of patients were greatly improved. The facial color indexes L, a, b values of the disease group at week 6 were different from those at week 0 (Pâ <â .05). The overall brightness and chroma of the patient's facial color were significantly improved. The patients had virtual string pulse at week 0, and mainly string I and string II at week 7. The pulse parameters t1, t5, w, w/t, h1, h5, h3/h1, and h5/h1 values at week 7 were different from those at weeks 0, 1, 2 (Pâ <â .05); the tongue image was mainly red and crimson, peeling or greasy fur at week 0, while at weeks 6, 7, mainly light red, or thin white tongue. The RSRN method can regulate the complexion, tongue and pulse condition, clinical symptoms of postmenopausal atherosclerosis.