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1.
Protein Cell ; 2024 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-38476024

RESUMO

Coactivator-associated arginine methyltransferase 1 (CARM1) promotes the development and metastasis of estrogen receptor alpha (ERα)-positive breast cancer. The function of CARM1 in triple-negative breast cancer (TNBC) is still unclear and requires further exploration. Here, we report that CARM1 promotes proliferation, epithelial-mesenchymal transition (EMT), and stemness in TNBC. CARM1 is upregulated in multiple cancers and its expression correlates with breast cancer progression. Genome-wide analysis of CARM1 showed that CARM1 is recruited by hypoxia-inducible factor 1 subunit alpha (HIF1A) and occupy the promoters of CDK4, Cyclin D1, ß-catenin, HIF1A, MALAT1, and SIX1 critically involved in cell cycle, HIF-1 signaling pathway, Wnt signaling pathway, VEGF signaling pathway, thereby modulating the proliferation and invasion of TNBC cells. We demonstrated that CARM1 is physically associated with and directly interacts with HIF1A. Moreover, we found that ellagic acid, an inhibitor of CARM1, can suppress the proliferation and metastasis of TNBC by directly inhibiting CDK4 expression. Our research has determined the molecular basis of CARM1 carcinogenesis in TNBC and its effective natural inhibitor, which may provide new ideas and drugs for cancer therapy.

3.
Cancer Commun (Lond) ; 43(10): 1117-1142, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37658635

RESUMO

BACKGROUND: Proteins containing the Jumonji C (JmjC) domain participated in tumorigenesis and cancer progression. However, the mechanisms underlying this effect are still poorly understood. Our objective was to investigate the role of Jumonji and the AT-rich interaction domain-containing 2 (JARID2) - a JmjC family protein - in breast cancer, as well as its latent association with obesity. METHODS: Immunohistochemistry, The Cancer Genome Atlas, Gene Expression Omnibus, and other databases were used to analyze the expression of JARID2 in breast cancer cells. Growth curve, 5-ethynyl-2-deoxyuridine (EdU), colony formation, and cell invasion experiments were used to detect whether JARID2 affected breast cancer cell proliferation and invasion. Spheroidization-based experiments and xenotumor transplantation in NOD/SCID mice were used to examine the association between JARID2 and breast cancer stemness. RNA-sequencing, Kyoto Encyclopedia of Genes and Genomes, and Gene Set Enrichment Analysis were used to identify the cell processes in which JARID2 participates. Immunoaffinity purification and silver staining mass spectrometry were conducted to search for proteins that might interact with JARID2. The results were further verified using co-immunoprecipitation and glutathione S-transferase (GST) pull-down experiments. Using chromatin immunoprecipitation (ChIP) sequencing, we sought the target genes that JARID2 and metastasis-associated protein 1 (MTA1) jointly regulated; the results were validated by ChIP-PCR, quantitative ChIP (qChIP) and ChIP-reChIP assays. A coculture experiment was used to explore the interactions between breast cancer cells and adipocytes. RESULTS: In this study, we found that JARID2 was highly expressed in multiple types of cancer including breast cancer. JARID2 promoted glycolysis, lipid metabolism, proliferation, invasion, and stemness of breast cancer cells. Furthermore, JARID2 physically interacted with the nucleosome remodeling and deacetylase (NuRD) complex, transcriptionally repressing a series of tumor suppressor genes such as BRCA2 DNA repair associated (BRCA2), RB transcriptional corepressor 1 (RB1), and inositol polyphosphate-4-phosphatase type II B (INPP4B). Additionally, JARID2 expression was regulated by the obesity-associated adipokine leptin via Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway in the breast cancer microenvironment. Analysis of various online databases also indicated that JARID2/MTA1 was associated with a poor prognosis of breast cancer. CONCLUSION: Our data indicated that JARID2 promoted breast tumorigenesis and development, confirming JARID2 as a target for cancer treatment.

4.
Adv Sci (Weinh) ; 10(14): e2202737, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36941223

RESUMO

Circadian rhythms, as physiological systems with self-regulatory functions in living organisms, are controlled by core clock genes and are involved in tumor development. The protein arginine methyltransferase 6 (PRMT6) serves as an oncogene in a myriad of solid tumors, including breast cancer. Hence, the primary aim of the current study is to investigate the molecular mechanisms by which the PRMT6 complex promotes breast cancer progression. The results show that PRMT6, poly(ADP-ribose) polymerase 1 (PARP1), and the cullin 4 B (CUL4B)-Ring E3 ligase (CRL4B) complex interact to form a transcription-repressive complex that co-occupies the core clock gene PER3 promoter. Moreover, genome-wide analysis of PRMT6/PARP1/CUL4B targets identifies a cohort of genes that is principally involved in circadian rhythms. This transcriptional-repression complex promotes the proliferation and metastasis of breast cancer by interfering with circadian rhythm oscillation. Meanwhile, the PARP1 inhibitor Olaparib enhances clock gene expression, thus, reducing breast carcinogenesis, indicating that PARP1 inhibitors have potential antitumor effects in high-PRMT6 expression breast cancer.


Assuntos
Neoplasias da Mama , Relógios Circadianos , Humanos , Feminino , Linhagem Celular Tumoral , Relógios Circadianos/genética , Transformação Celular Neoplásica , Núcleo Celular/metabolismo , Neoplasias da Mama/metabolismo , Proteínas Nucleares/genética , Proteína-Arginina N-Metiltransferases/genética , Proteína-Arginina N-Metiltransferases/metabolismo , Poli(ADP-Ribose) Polimerase-1/genética , Proteínas Culina/genética
5.
BMC Musculoskelet Disord ; 23(1): 455, 2022 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-35568888

RESUMO

BACKGROUND: Based on the Digital Imaging and Communications in Medicine (DICOM) data of three-dimensional magnetic resonance imaging (3D-MRI), finite element models of the hip joints of children with developmental dysplasia of the hip were established. The primary objectives included simulation and analysis of the finite element model pre- and post-closed reduction under different stances and loads, and evaluation of the size and distribution of von Mises stress in the acetabulum and femoral head pre- and post-operation and the short-term effects. METHODS: Acetabular index measurements of both the unaffected and affected sides were conducted, alongside International Hip Dysplasia Institute (IHDI) classification of the affected hip. Establishing the finite element model of both the affected and unaffected hips was based on the 3D-T1WI sequence DICOM data, using Mimics, 3-matic, and Ansys software, before and after closed reduction surgery. The size and distribution data of von Mises stress on the affected side of the acetabulum and femoral head were collected pre- and post-operation. RESULTS: The study indicated that the increasing acetabular index of the affected hip was directly proportional to the increasing severity based on IHDI classification (P < 0.05). Preoperative IHDI classification significantly correlated with the von Mises stress (r = 0.560-0.569, 0.562-0.564, P < 0.05). Under different stances and load conditions, the von Mises stress on the affected side post-operation was lower than that noted pre-operation (P < 0.01), while that on the acetabulum increased proportionally to the load. Although the magnitude and distribution of von Mises stress on the affected side of the acetabulum were similar to those on the healthy side post-operation, there were statistical differences between the two (P < 0.01). The von Mises stress of the lateral column of the femoral head post-operation was significantly lower than that noted pre-operation (P < 0.01). While the high-stress points of the lateral column disappeared post-operation, the von Mises stress was evenly distributed in the femoral head. CONCLUSIONS: The 3D-MRI finite element could provide the von Mises stress value and distribution characteristics of the acetabulum and femoral head pre- and post-operation. Closed reduction can, therefore, improve the size and distribution of von Mises stress on the affected acetabulum and femoral head.


Assuntos
Displasia do Desenvolvimento do Quadril , Luxação Congênita de Quadril , Acetábulo/diagnóstico por imagem , Acetábulo/cirurgia , Criança , Análise de Elementos Finitos , Luxação Congênita de Quadril/diagnóstico por imagem , Luxação Congênita de Quadril/cirurgia , Humanos , Imageamento por Ressonância Magnética
6.
Breast Cancer ; 29(4): 645-658, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35174450

RESUMO

BACKGROUND: RNA N6-methyladenosine (m6A) modification is primarily regulated by m6A regulators, which play significant epigenetic regulatory roles in tumorigenesis, tumor development, and tumor immune microenvironment. However, the correlation between m6A regulators and immune cell infiltration in breast cancer remains unclear. METHODS: In this study, m6A modification patterns were evaluated based on 31 m6A modification regulators. m6A clusters were determined by consensus clustering. Immune landscape and immune cell infiltration subgroups were characterized by m6A clusters. Key module and hub genes related to m6A regulators and immune infiltration cells were identified by WGCNA. LASSO algorithm was applied to select prognostic signatures. Multivariate Cox regression analysis was applied to assess the prognostic value of gene signatures. RESULTS: Two distinct m6A clusters were determined based on the expression of 31 m6A modification regulators and characterized by two tumor immune microenvironment (TIME) immune cell infiltration subgroups. Further, a total of 1971 differentially expressed genes between breast cancer patients and healthy controls were screened, nine modules associated with clinical characteristics of breast cancer patients were identified. Later, one key module and 13 hub genes correlated with m6A regulators and immune infiltration cells were identified. LASSO Cox regression analysis selected and constructed a ten-gene prognostic model to build a risk score system for individual breast cancer patient prognosis. The performance of the ten-gene-based risk score system was further validated in an independent dataset with an AUC of 0.659. CONCLUSIONS: This study revealed that m6A modification regulators played a significant role in the TIME regulation of breast cancer. The hub ten gene-based risk score system is valuable in predicting the prognosis of breast cancer patients, which may provide potential significance for breast cancer diagnosis, prognosis, and immunotherapy in the future.


Assuntos
Neoplasias da Mama , Biomarcadores Tumorais/genética , Neoplasias da Mama/genética , Feminino , Humanos , Metilação , RNA , Fatores de Risco , Microambiente Tumoral/genética
7.
J Neurosci Methods ; 371: 109498, 2022 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-35167839

RESUMO

Due to its advantages of high temporal and spatial resolution, the technology of simultaneous electroencephalogram-functional magnetic resonance imaging (EEG-fMRI) acquisition and analysis has attracted much attention, and has been widely used in various research fields of brain science. However, during the fMRI of the brain, ballistocardiogram (BCG) artifacts can seriously contaminate the EEG. As an unpaired problem, BCG artifact removal now remains a considerable challenge. Aiming to provide a solution, this paper proposed a novel modular generative adversarial network (GAN) and corresponding training strategy to improve the network performance by optimizing the parameters of each module. In this manner, we hope to improve the local representation ability of the network model, thereby improving its overall performance and obtaining a reliable generator for BCG artifact removal. Moreover, the proposed method does not rely on additional reference signal or complex hardware equipment. Experimental results show that, compared with multiple methods, the technique presented in this paper can remove the BCG artifact more effectively while retaining essential EEG information.


Assuntos
Artefatos , Balistocardiografia , Algoritmos , Balistocardiografia/métodos , Eletroencefalografia/métodos , Imageamento por Ressonância Magnética/métodos
8.
Cell Death Differ ; 28(12): 3329-3343, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34163012

RESUMO

Pancreatic cancer is a common malignant tumor with poor prognosis. Recently, cancer stem cells (CSCs) were identified in several solid tumors, including pancreatic cancer. Although accumulating evidence indicates that sirtuin 1 (SIRT1) exerts biological functions in various cancers, how it contributes to tumorigenesis and metastasis of pancreatic cancer, as well as its role in CSCs, is still poorly defined. Here we show that SIRT1 interacts with the Cullin 4B (CUL4B)-Ring E3 ligase (CRL4B) complex, which is responsible for H2AK119 monoubiquitination (H2AK119ub1), collaborating as a functional unit. Genome-wide analysis of SIRT1/CUL4B targets identified a cohort of genes, including GRHL3 and FOXO3, critically involved in cell differentiation, growth, and migration. Furthermore, we found that SIRT1 and CUL4B collectively promote the proliferation, autophagy, and invasion of pancreatic cancer cells. Remarkably, we demonstrate that SIRT1/CUL4B promotes CSC-like properties, including increased stemness marker expression and sphere formation. In vivo experiments implied that SIRT1 promoted established tumor xenograft growth, increased tumor-initiating capacity in NOD/SCID mice, and increased CSC frequency. Strikingly, SIRT1 and CUL4B expression is markedly upregulated in a variety of human cancers, including pancreatic cancer. Our data provide a molecular basis for the functional interplay between histone deacetylation and ubiquitination. The results also implicate the SIRT1/CRL4B complex in pancreatic cancer metastasis and stem cell properties, thus supporting SIRT1 as a promising potential target for cancer therapy development.


Assuntos
Células-Tronco Neoplásicas/metabolismo , Neoplasias Pancreáticas/genética , Receptores de Interleucina-17/metabolismo , Sirtuína 1/metabolismo , Animais , Carcinogênese , Humanos , Camundongos , Neoplasias Pancreáticas/patologia , Transfecção
9.
Curr Med Imaging ; 17(8): 1010-1017, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33573574

RESUMO

OBJECTIVE: To compare conventional sensitivity encoding turbo spin-echo (SENSE-TSE) with compressed sensing plus SENSE turbo spin-echo (CS-TSE) in lumbar vertebrae magnetic resonance imaging (MRI). METHODS: This retrospective study of lumbar vertebrae MRI included 600 patients; 300 patients received SENSE-TSE and 300 patients received CS-TSE. The SENSE acceleration factor was 1.4 for T1WI, 1.7 for T2WI, and 1.7 for PDWI. The CS total acceleration factor was 2.4, 3.6, 4.0, and 4.0 for T1WI, T2WI, PDWI sagittal, and T2WI transverse, respectively. The image quality of each MRI sequence was evaluated objectively by the signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) and subjectively on a five-point scale. Two radiologists independently reviewed the MRI sequences of the 300 patients receiving CS-TSE, and their diagnostic consistency was evaluated. The degree of intervertebral foraminal stenosis and nerve root compression was assessed using the T1WI sagittal and T2WI transverse images. RESULTS: The scan time was reduced from 7 min 28 s to 4 min 26 s with CS-TSE. The median score of nerve root image quality was 5 (p > 0.05). The diagnostic consistency using CS-TSE images between the two radiologists was high for diagnosing lumbar diseases (κ > 0.75) and for evaluating the degree of lumbar foraminal stenosis and nerve root compression (κ = 0.882). No differences between SENSE-TSE and CS-TSE were observed for sensitivity, specificity, positive predictive value, or negative predictive value. CONCLUSION: CS-TSE has the potential for diagnosing lumbar vertebrae and disc disorders.


Assuntos
Imageamento Tridimensional , Vértebras Lombares , Humanos , Vértebras Lombares/diagnóstico por imagem , Imageamento por Ressonância Magnética , Estudos Retrospectivos , Razão Sinal-Ruído
10.
Acta Radiol ; 62(10): 1418-1425, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33108893

RESUMO

BACKGROUND: T2 mapping is useful for evaluating the cartilage matrix. PURPOSE: To determine the variations in the acetabular cartilage T2 relaxation values between healthy individuals and those with developmental dysplasia of the hip (DDH). MATERIAL AND METHODS: Thirty-three patients with unilateral DDH underwent 3-T magnetic resonance imaging (MRI) between January 2018 and February 2019. Fifteen volunteers (30 hips) were enrolled as controls. T2 values were measured with the T2 mapping sequence in all layers and were equally divided into three layers (deep, middle, and superficial) with equal thickness. We calculated the mean T2 relaxation values for the full thickness, deep, middle, and superficial layers and compared the values between the different groups. In addition, the inter- and intra-observer agreements were calculated. RESULTS: The T2 relaxation values in the DDH arm were significantly lower in the middle, superficial, and full thickness layers compared with those of the volunteers and contralateral hips. The T2 relaxation values of the deep layers showed no significant difference between the different groups. The acetabular cartilage T2 relaxation values increased from the deep layer to the superficial layer in the control and contralateral groups. Both inter- and intra-observer agreements were good. CONCLUSION: MRI T2 mapping may help to diagnose developmental disorders of the acetabular cartilage matrix in infants and children with DDH. Abnormal acetabular cartilage T2 relaxation values may be due to the extraordinary stress load of the femoral head.


Assuntos
Cartilagem/diagnóstico por imagem , Cartilagem/fisiopatologia , Displasia do Desenvolvimento do Quadril/diagnóstico por imagem , Displasia do Desenvolvimento do Quadril/fisiopatologia , Imageamento por Ressonância Magnética/métodos , Acetábulo/diagnóstico por imagem , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Estudos Prospectivos
11.
Cell Rep ; 29(6): 1482-1498.e4, 2019 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-31693890

RESUMO

The histone methyl transferase enhancer of zeste homolog 2 (EZH2) is a master transcriptional regulator involved in histone H3 lysine 27 trimethylation. We aimed to elucidate the precise post-translational regulations of EZH2 and their role in cancer pathogenesis. Here, we show that SET and MYND domain containing 2 (SMYD2) directly methylates EZH2 at lysine 307 (K307) and enhances its stability, which can be relieved by the histone H3K4 demethylase lysine-specific demethylase 1 (LSD1). SMYD2 is critical for EZH2 function in repressing a cohort of genes governing several cancer-associated pathways. In addition, SMYD2 promotes breast cancer cell proliferation, epithelial-mesenchymal transition, and invasion through EZH2 K307 methylation, and it is markedly upregulated in various human cancers. Our data suggest that dynamic crosstalk between SMYD2-mediated EZH2 methylation plays an important role in fine-tuning EZH2 functions in chromatin recruitment and transcriptional repression.


Assuntos
Neoplasias da Mama/metabolismo , Carcinogênese/genética , Cromatina/metabolismo , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Histona-Lisina N-Metiltransferase/metabolismo , Lisina/metabolismo , Neoplasias da Mama/genética , Neoplasias da Mama/mortalidade , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Cromatina/genética , Imunoprecipitação da Cromatina , Bases de Dados Genéticas , Progressão da Doença , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Histona Desmetilases/genética , Histona Desmetilases/metabolismo , Histona-Lisina N-Metiltransferase/genética , Humanos , Metilação , Complexo Repressor Polycomb 2/genética , Complexo Repressor Polycomb 2/metabolismo , Processamento de Proteína Pós-Traducional
12.
J Biol Chem ; 294(43): 15808-15825, 2019 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-31492753

RESUMO

GATA3 is a basic and essential transcription factor that regulates many pathophysiological processes and is required for the development of mammary luminal epithelial cells. Loss-of-function GATA3 alterations in breast cancer are associated with poor prognosis. Here, we sought to understand the tumor-suppressive functions GATA3 normally performs. We discovered a role for GATA3 in suppressing epithelial-to-mesenchymal transition (EMT) in breast cancer by activating miR-455-3p expression. Enforced expression of miR-455-3p alone partially prevented EMT induced by transforming growth factor ß (TGF-ß) both in cells and tumor xenografts by directly inhibiting key components of TGF-ß signaling. Pathway and biochemical analyses showed that one miRNA-455-3p target, the TGF-ß-induced protein ZEB1, recruits the Mi-2/nucleosome remodeling and deacetylase (NuRD) complex to the promotor region of miR-455 to strictly repress the GATA3-induced transcription of this microRNA. Considering that ZEB1 enhances TGF-ß signaling, we delineated a double-feedback interaction between ZEB1 and miR-455-3p, in addition to the repressive effect of miR-455-3p on TGF-ß signaling. Our study revealed that a feedback loop between these two axes, specifically GATA3-induced miR-455-3p expression, could repress ZEB1 and its recruitment of NuRD (MTA1) to suppress miR-455, which ultimately regulates TGF-ß signaling. In conclusion, we identified that miR-455-3p plays a pivotal role in inhibiting the EMT and TGF-ß signaling pathway and maintaining cell differentiation. This forms the basis of that miR-455-3p might be a promising therapeutic intervention for breast cancer.


Assuntos
Células Epiteliais/metabolismo , Fator de Transcrição GATA3/metabolismo , MicroRNAs/metabolismo , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo , Animais , Sequência de Bases , Mama/patologia , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Proliferação de Células/genética , Transição Epitelial-Mesenquimal/genética , Receptor alfa de Estrogênio/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Células MCF-7 , Camundongos SCID , MicroRNAs/genética , Metástase Neoplásica , Proteínas de Neoplasias/metabolismo , Transcrição Gênica , Homeobox 1 de Ligação a E-box em Dedo de Zinco/metabolismo
13.
Eur Cytokine Netw ; 30(2): 59-66, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31486397

RESUMO

Recent studies have demonstrated that nicotine exhibited anti-inflammatory and neuroprotective properties by interacting with the alpha 7 nicotinic acetylcholine receptor (α7nAChR). However, the role of nicotine in regeneration during peripheral nerve injury has not been elucidated. The aim of this study was to investigate whether nicotine down-regulated production of proinflammatory cytokines and promoted peripheral nerve regeneration in rats. Rats challenged with sciatic nerve crush injury were treated with nicotine (1.5 mg/kg), three times per day. The expression of the proinflammatory cytokines tumor necrosis factor alpha (TNF-α) and interleukin (IL-1ß), pinch test results, growth-associated protein 43 (GAP-43) expression, morphometric analyses, and the sciatic functional indexes were determined in sciatic nerves. Treatment with nicotine decreased local levels of TNF-α and IL-1ß, and increased the expression of GAP-43. Nicotine also improved nerve regeneration and functional recovery. The overall protective effects of nicotine were reversed by concomitant treatment with α7nACHR antagonist methyllycaconitine, indicating that nicotine exerted its specific anti-inflammatory and neuroprotective effects through the α7nAChR. These findings show that nicotine administration can provide a potential therapeutic pathway for the treatment of peripheral nerve injury, by a direct protective effect through the α7nAChR-mediated cholinergic anti-inflammatory pathway.


Assuntos
Lesões por Esmagamento/metabolismo , Lesões por Esmagamento/patologia , Citocinas/metabolismo , Mediadores da Inflamação/metabolismo , Fármacos Neuroprotetores/farmacologia , Nicotina/farmacologia , Nervo Isquiático/metabolismo , Nervo Isquiático/patologia , Animais , Modelos Animais de Doenças , Proteína GAP-43/metabolismo , Interleucina-1beta/metabolismo , Masculino , Atividade Motora/efeitos dos fármacos , Regeneração Nervosa/efeitos dos fármacos , Ratos Wistar , Nervo Isquiático/efeitos dos fármacos , Células Receptoras Sensoriais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo
14.
Anal Chem ; 91(13): 7980-7984, 2019 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-31247714

RESUMO

Surface-enhanced Raman spectroscopy (SERS) has exhibited great potential in label-free DNA detection. Owing to the limitation in chain length, it is however still challenging for SERS as a routine method to explore the intrinsic structural information on unmodified DNA. Here, we develop a universal SERS-based approach toward quantification of A/G in single-stranded DNAs (12 up to 28 bases) by introducing a novel interfacial agent, dichloromethane. DNA hybridization is successfully probed as evidenced by the typical SERS bands attributed to hydrogen bonds in a hairpin structure. More importantly, enlarged space of "hot spots" in SERS enables discrimination of single-base mutation in double-stranded DNA with 100 bases, which as a proof-of-concept study will pave a new avenue for highly sensitive DNA detection in clinical applications.


Assuntos
DNA/genética , Mutação Puntual , Análise Espectral Raman/métodos , Sequência de Bases , DNA/análise , DNA de Cadeia Simples/análise , DNA de Cadeia Simples/genética , Indicadores e Reagentes , Cloreto de Metileno , Modelos Moleculares , Conformação de Ácido Nucleico , Hibridização de Ácido Nucleico/métodos
15.
J Phys Chem Lett ; 10(11): 3013-3018, 2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-31091107

RESUMO

Direct, label-free sequence analysis of DNA hybridization has been achieved by surface-enhanced Raman spectroscopy. In this work, aluminum-ion-aggregated and iodide-modified silver nanoparticles were used as substrates to obtain Raman spectra of the DNA strands with the same base composition but different sequences, which form random coils or various hairpin conformations. Upon DNA hybridization, reproducibly enhanced bands were easily observed, corresponding well to the formation of Watson-Crick hydrogen bonds, base ring breathing vibrations, and hairpin loops. These characteristic bands can be used to unambiguously distinguish the hairpins from the random DNA conformation. Moreover, by using the deoxyribose band (959 cm-1) as an internal standard to normalize the characteristic bands at 1703 cm-1 corresponding to the dG νC=O H bond, the guanine-cytosine base-pair contents and sequence in DNA hairpins can be accurately measured. Applying this method, a single base mutation in a functional double helix was confidently identified.


Assuntos
DNA/química , Alumínio/química , Composição de Bases , Pareamento Incorreto de Bases , Pareamento de Bases , Sequência de Bases , Citosina/química , Guanina/química , Ligação de Hidrogênio , Iodetos/química , Nanopartículas Metálicas/química , Tamanho da Partícula , Prata/química , Análise Espectral Raman , Propriedades de Superfície
16.
J Am Soc Mass Spectrom ; 30(4): 625-633, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30747410

RESUMO

In the present work, we developed an UV-absorptive and highly reactive tag aromatic molecule, quinoline-5,8-dione (QLD), for robust quantitative analysis of GSH by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The QLD could react with GSH with high efficiency at room temperature, and the resultant QLD-GSH conjugate could be readily detected by MALDI MS without interferences. By using the QLD tag, the detection limit of GSH was lowered to 10 fmol µL-1, which was four orders of magnitude higher than that detected without using the QLD tag. Furthermore, accurate quantitative measurements of GSH in solution were successfully demonstrated by using glutamic acid-cysteine-alanine (ECA) as an internal standard. By properly adjusting the ECA concentrations, the intensity ratio value of QLD-tagged GSH (QLD-GSH) to QLD-tagged ECA (QLD-ECA) displayed a good linearity with GSH concentrations in a broad range from 4 to 4000 µM. Finally, the GSH level in HeLa cell lysates was also successfully detected, and the results are consistent with that obtained by a colorimetric assay. In summary, the proposed QLD-based tag method should be a rapid, cost-/time-effective, and sensitive new method for quantitative determination of GSH by MALDI MS. Graphical Abstract.

17.
Langmuir ; 34(50): 15388-15395, 2018 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-30461285

RESUMO

In the present work, an interesting droplet self-retraction phenomenon of myristyltrimethylammonium bromide (C14TAB) solution on the superhydrophilic mica surface was observed. The self-retraction could only occur within a concentration range from 0.01 to 16 cmc. The maximum variation of the contact angle (from 24.99 to 76.85°) was observed in droplets with a concentration of 0.1 cmc. The self-retraction mechanism was studied based on high-speed photography, surface analyses, surface energy calculation, and model fitting. It was proved that there was a monolayer of C14TAB molecules adsorbed on the mica surface, which formed through the electrostatic interaction between the negative mica and the positively charged headgroups. The formation of this monolayer took only a short time of <67 ms since the droplet touched the surface, and the monolayer was well-distributed and hydrophobic with a surface free energy of 76.041 mJ/m2 (20% of mica's surface free energy). The reduction of surface tension led to an imbalance at the borderline of the droplet, causing self-retraction. A model was established to simulate the self-retraction process and it agreed with the experimental data well. This study has significant implication in understanding the surface modification caused by surfactant molecules.

18.
Eur J Obstet Gynecol Reprod Biol ; 228: 38-42, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29908376

RESUMO

OBJECTIVE: This study was to determinate the expression of Tspan5 in tubal ectopic implantation sites and to explore the correlation of the expressive level of Tspan5 at maternal-fetal interface and the occurrence of tubal ectopic pregnancy. STUDY DESIGN: This is a retrospective study. Trophoblastic and endometrial tissues were collected from tubal ectopic pregnancy(Total of 40), and intrauterine pregnancy(Total of 41), who had voluntary abortion, non-pregnancy women(Total of 12), who recieved an diagnostic uterine curettage before IVF-ET for male infertility. All samples were collected from women aged 23-40 years, from February 2012 to January 2014. Results 1. In human villi Tspan5 was primarily located in cytoplasm and on the surfaces of cytotroblasts(CTs) and extravillous trophoblast(EVCTs). The intensity of Tspan5 in tubal pregnancy was significantly higher than that in normal intrauterine pregnancy, showing significant differences (Mean of IOD:109.39 ±â€¯61.84 Vs. 89.04 ±â€¯36.44;t = 2.33, P = 0.023). 2. In human deciduas of intrauterine pregnancy or endometrium of tubal pregnancy and non-pregnancy Tspan5 expressed in cytoplasm and membrane of glandular epithelial cells. The expressive level of this protein was increased in tubal pregnancy than that in intrauterine pregnancy and non-pregnancy(Mean of IOD:144.18 ±â€¯106.22 Vs. 93.43 ±â€¯67.10, P = 0.037; 144.18 ±â€¯106.22 Vs. 88.56 ±â€¯33.24, P = 0.018). CONCLUSION: Our study indicated that the trophoblasts in tubal pregnancy showed more proliferative and invasive characteristics. Dysregulation of Tspan5 in decidual microenvironment may relate to the retention of embryo in fallopian tube. SUPPORT: This study was Supported by Science and Technology Planning Project of Guangdong Province.


Assuntos
Vilosidades Coriônicas/metabolismo , Gravidez Tubária/metabolismo , Tetraspaninas/metabolismo , Trofoblastos/fisiologia , Adulto , Proliferação de Células , Endométrio/metabolismo , Feminino , Humanos , Gravidez , Estudos Retrospectivos , Adulto Jovem
19.
Adv Sci (Weinh) ; 5(3): 1700616, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29593965

RESUMO

2D or 3D layered materials, such as graphene, graphite, and molybdenum disulfide, usually exhibit superlubricity properties when sliding occurs between the incommensurate interface lattices. This study reports the superlubricity between graphite and silica under ambient conditions, induced by the formation of multiple transferred graphene nanoflakes on the asperities of silica surfaces after the initial frictional sliding. The friction coefficient can be reduced to as low as 0.0003 with excellent robustness and is independent of the surface roughness, sliding velocities, and rotation angles. The superlubricity mechanism can be attributed to the extremely weak interaction and easy sliding between the transferred graphene nanoflakes and graphite in their incommensurate contact. This finding has important implications for developing approaches to achieve superlubricity of layered materials at the nanoscale by tribointeractions.

20.
Chin J Nat Med ; 15(5): 384-391, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28558874

RESUMO

The fruits of Paulownia catalpifolia Gong Tong are used as a Chinese folk herbal medicine for the treatment of enteritis, tonsillitis, bronchitis, and dysentery, etc. Our previous study has identified new C-geranylated flavanones with obvious anti-proliferative effects in lung cancer A549 cells. In the present study, a new C-geranylated flavone, paucatalinone C (1) and five known C-geranylated flavanones (2-6) were isolated. In addition, a total of 34 C-geranylated flavonoids were detected by HPLC-DAD-ESI-MS/MS coupling techniques from the CH2Cl2 extract of P. catalpifolia. Futhermore, anti-aging effects of isolated compounds were evaluated in vitro with premature senescent 2BS cells induced by H2O2. Phytochemical results indicated that P. catalpifolia was a natural resource of abundant C-geranylated flavonoids. Diplacone (3) and paucatalinone A (5) were the potent anti-aging agents in the premature senescent 2BS cells induced by H2O2 and the C-geranyl substituent may be an important factor because of its lipophilic character.


Assuntos
Envelhecimento/efeitos dos fármacos , Flavonoides/química , Flavonoides/farmacologia , Magnoliopsida/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Flavonoides/isolamento & purificação , Frutas/química , Humanos , Peróxido de Hidrogênio/toxicidade , Estrutura Molecular , Extratos Vegetais/isolamento & purificação , Espectrometria de Massas em Tandem
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