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1.
Biomed Res Int ; 2021: 4873678, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34337013

RESUMO

LIHC (liver hepatocellular carcinoma) mostly occurs in patients with chronic liver disease. It is primarily induced by a vicious cycle of liver injury, inflammation, and regeneration that usually last for decades. The G protein nucleolar 2 (GNL2), as a protein-encoding gene, is also known as NGP1, Nog2, Nug2, Ngp-1, and HUMAUANTIG. Few reports are shown towards the specific biological function of GNL2. Meanwhile, it is still unclear whether it is related to the pathogenesis of carcinoma up to date. Here, our study attempts to validate the role and function of GNL2 in LIHC via multiple databases and functional assays. After analysis of gene expression profile from The Cancer Genome Atlas (TCGA) database, GNL2 was largely heightened in LIHC, and its overexpression displayed a close relationship with different stages and poor prognosis of carcinoma. After enrichment analysis, the data revealed that the genes coexpressed with GNL2 probably participated in ribosome biosynthesis which was essential for unrestricted growth of carcinoma. Cell functional assays presented that GNL2 knockdown by siRNA in LIHC cells MHCC97-H and SMCC-7721 greatly reduced cell proliferation, migration, and invasion ability. All in all, these findings capitulated that GNL2 could be a promising treatment target and prognosis biomarker for LIHC.


Assuntos
Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/terapia , Proteínas de Ligação ao GTP/metabolismo , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/terapia , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Regulação para Baixo/genética , Proteínas de Ligação ao GTP/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Ontologia Genética , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Prognóstico , Reprodutibilidade dos Testes , Transdução de Sinais/genética
2.
Biosens Bioelectron ; 191: 113455, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34175650

RESUMO

A novel 3D CdSe quantum dots (QDs)-DNA nanonetwork was assembled to sensitize the mesoporous SnO2 photoelectrochemical (PEC) substrate, which was coupled with a biped-DNA walker multiple amplification technique to design a versatile electrochemiluminescence (ECL) and PEC biosensor for dual detection of HIV. Firstly, the photosensitive CdSe QDs and SnO2 nanoflowers have well-matched band-edge energy level, thus their complex can promote effective transfer of the photogenerated carriers, and show better PEC and ECL property. Then, a novel 3D CdSe QDs-DNA nanonetwork was assembled and loaded with a large amount of QDs, which was used as multifunctional PEC and ECL probes. Moreover, the target-triggered biped DNA walker-cascade amplification method was introduced to generate a large amount of output DNA, which was used to link numerous 3D CdSe QDs-DNA nanonetwork probes to the electrode, generating greatly amplified signals for sensitive assay of HIV. The highly photosensitive 3D CdSe QDs-DNA reticulated nanomaterials have high stability and controllability, and display significantly improved PEC and ECL signals of the biosensor. This method opened a new photoelectric nanocomposite of QDs-sensitized SnO2 nanoflower, and developed a versatile biosensing strategy using the 3D CdSe QDS DNA sensitization probes for ultra-sensitive detection of biomolecules, which is important for the early diagnosis of diseases.


Assuntos
Técnicas Biossensoriais , Compostos de Cádmio , Infecções por HIV , Pontos Quânticos , Compostos de Selênio , DNA , Técnicas Eletroquímicas , Infecções por HIV/diagnóstico , Humanos , Medições Luminescentes
3.
Int J Mol Sci ; 22(9)2021 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-34066572

RESUMO

The NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER FAMILY (NPF) genes, initially characterized as nitrate or peptide transporters in plants, are involved in the transport of a large variety of substrates, including amino acids, nitrate, auxin (IAA), jasmonates (JAs), abscisic acid (ABA) and gibberellins (GAs) and glucosinolates. A total of 169 potential functional NPF genes were excavated in Brassica napus, and they showed diversified expression patterns in 90 different organs or tissues based on transcriptome profile data. The complex time-serial expression changes were found for most functional NPF genes in the development process of leaves, silique walls and seeds, which indicated that the expression of Brassica napus NPF (BnaNPF) genes may respond to altered phytohormone and secondary metabolite content through combining with promoter element enrichment analysis. Furthermore, many BnaNPF genes were detected to respond to vernalization with two different patterns, and 20 BnaNPF genes responded to nitrate deficiency. These results will provide useful information for further investigation of the biological function of BnaNPF genes for growth and development in rapeseed.


Assuntos
Proteínas de Transporte de Ânions/genética , Brassica napus/genética , Brassica napus/fisiologia , Flores/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Nitrogênio/deficiência , Proteínas de Plantas/genética , Sequência de Aminoácidos , Proteínas de Transporte de Ânions/química , Proteínas de Transporte de Ânions/metabolismo , Brassica napus/efeitos dos fármacos , Variações do Número de Cópias de DNA/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Nitratos/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Domínios Proteicos , Especificidade da Espécie , Sintenia/genética
4.
Nanoscale ; 13(16): 7678-7684, 2021 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-33928980

RESUMO

In this paper, a novel photoelectrochemical (PEC) "signal-on" biosensor based on a Bi2Sn2O7/Bi2S3 heterojunctioncoupled with target-switchable DNA hydrogels is reported for the ultrasensitive detection of P53 gene DNA. For the first time, sulfide ions are discovered to display an excellent PEC sensitization effect on Bi2Sn2O7 material by forming the Bi2Sn2O7/Bi2S3 heterojunction. The sensitization amplitude increased by 63 times, and the photocurrent polarity switched from cathodic to anodic. When the target DNA-induced-cycling amplification process produced a mass of product chains (PD), PD was introduced into the target-switchable DNA hydrogels to quantitatively release sulfide ions, which were further introduced to the Bi2Sn2O7-modified PEC platform and resulted in an enormous enhancement of the PEC signal due to the significant sensitization effect of sulfide ions on Bi2Sn2O7via an anion-exchange reaction. The corresponding PEC signal change of the Bi2Sn2O7/Bi2S3 platform was used for the detection of target DNA. This biosensing strategy opens up a novel sulfide ion-sensitized PEC platform and exhibits excellent analytical performance with a wide linear range (100 fM-10 nM), which has broad application prospects in bioanalysis and clinical diagnosis.


Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Ânions , DNA , Hidrogéis , Limite de Detecção
5.
Ann Hematol ; 100(6): 1547-1552, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33839882

RESUMO

POEMS syndrome is a rare plasma cell disorder. Lenalidomide has recently emerged as a therapeutic option for POEMS syndrome. Cereblon has been identified as the direct target of lenalidomide, and high cereblon expression is associated with better response and outcome to lenalidomide therapy in multiple myeloma patients. Here, we analyzed the predictive value of cereblon, IKZF1, and IKZF3 in CD138+ selected plasma cells from forty-one newly diagnosed POEMS syndrome patients treated with lenalidomide in combination with dexamethasone at both gene and protein levels. We found that patients with high cereblon expression tended to achieve better hematologic response compared to those with low expression (p = 0.024 for gene expression; p = 0.01 for protein expression). Multivariate Cox regression analysis revealed high cereblon mRNA expression as an independent prognostic marker for longer progression-free survival (hazard ratio 0.542; 95% CI 0.337-0.871; p = 0.011). In conclusion, our results emphasized the role of cereblon mRNA expression as a unique biomarker for predicting the clinical response and outcome of lenalidomide-based therapy in newly diagnosed POEMS syndrome patients.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Anti-Inflamatórios/uso terapêutico , Dexametasona/uso terapêutico , Fatores Imunológicos/uso terapêutico , Lenalidomida/uso terapêutico , Síndrome POEMS/tratamento farmacológico , Ubiquitina-Proteína Ligases/genética , Adulto , Idoso , Biomarcadores Tumorais/genética , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Síndrome POEMS/diagnóstico , Síndrome POEMS/genética , Prognóstico , Estudos Prospectivos , RNA Mensageiro/genética , Resultado do Tratamento , Regulação para Cima/efeitos dos fármacos , Adulto Jovem
6.
Artigo em Inglês | MEDLINE | ID: mdl-33560983

RESUMO

Among tracking techniques applied in the 3-D freehand ultrasound (US), the camera-based tracking method is relatively mature and reliable. However, constrained by manufactured marker rigid bodies, the US probe is usually limited to operate within a narrow rotational range before occlusion issues affect accurate and robust tracking performance. Thus, this study proposed a hemispherical marker rigid body to hold passive noncoplanar markers so that the markers could be identified by the camera, mitigating self-occlusion. The enlarged rotational range provides greater freedom for sonographers while performing examinations. The single-axis rotational and translational tracking performances of the system, equipped with the newly designed marker rigid body, were investigated and evaluated. Tracking with the designed marker rigid body achieved high tracking accuracy with 0.57° for the single-axis rotation and 0.01 mm for the single-axis translation for sensor distance between 1.5 and 2 m. In addition to maintaining high accuracy, the system also possessed an enhanced ability to capture over 99.76% of the motion data in the experiments. The results demonstrated that with the designed marker rigid body, the missing data were remarkably reduced from over 15% to less than 0.5%, which enables interpolation in the data postprocessing. An imaging test was further conducted, and the volume reconstruction of a four-month fetal phantom was demonstrated using the motion data obtained from the tracking system.

7.
Anal Chim Acta ; 1148: 238169, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33516380

RESUMO

In this work, a new fluorescence biosensor platform based on distance-dependent photoinduced-electron transfer (PET) coupled with target cross-chain displacement cyclic amplification strategy was developed to detect MicroRNA. The DNA cross structure was cleverly designed to protect restriction site, then multiple amplification reactions of target cycle and chain replacement based on DNA cross-configuration were carried out in the presence of primer, polymerase and cutting enzyme, thus a large number of single-stranded (ss) DNA products (S1 and S2) can be exported by inputting a small amount of target miRNA. The fluorescent AgNCs/DNA probe was synthesized based on high affinity of Ag to cytosine (C) rich in ssDNA acting as electron donor, and guanine (G) rich ssDNA can form G-quadruplex complex acting as electron receptor to induce PET process. S1 and S2 hybridized with flexible single-stranded DNA COM 1 and Com 2, forming rigid double-stranded DNA to inhibit fluorescence quenching PET process, so the corresponding fluorescence was recovered. Thus the miRNA-induced amplified products can specifically result in fluorescence changes by PET, and the changes increase with increasing miRNA concentration. Therefore, the proposed fluorescent biosensor can be applied to quantitative determination of miRNA-182-5p, which has great potential in early clinical diagnosis of miRNAs related diseases.


Assuntos
Técnicas Biossensoriais , MicroRNAs , DNA/genética , Elétrons , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico
8.
Anal Chim Acta ; 1148: 338201, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33516383

RESUMO

As an important protease, trypsin (TRY) has been identified as a key indicator of various diseases. A simple and sensitive strategy for TRY detection by using an environment-friendly biosafe probe is significant. Herein, we introduced negatively charged fluorescent polydopamine nanoparticles (PDNPs) with 4.8 nm diameter obtained through a controllable method as an effective probe for TRY. PDNPs exhibited excellent fluorescence property but integrated with protamine (Pro) to form an aggregation-caused quenching system via a static quenching mechanism. The quenching mechanism of Pro to PDNPs revealed the significant effect of the surface charge, functional groups, and appropriate size of PDNPs on quenching process. Given the specific hydrolysis of Pro by TRY, PDNPs were released from the quenching integration of PDNPs and Pro (PDNPs-Pro) and recovered their fluorescence. Thus, a fluorescence sensor for TRY with a linear range of 0.01 and 0.1 µg/mL and a detection limit of 6.7 ng/mL was developed without the disturbing from other proteases. Compared with other TRY assays, the biosensor based on PDNPs-Pro has the advantages of simple operation, environmental friendliness, and high sensitivity. This specific controlled-synthesis PDNPs would open up a new window for the extended application of fluorescent nanomaterials in biomedicine based on fluorescence changes induced by biological interaction.


Assuntos
Nanopartículas , Protaminas , Indóis , Polímeros , Tripsina
9.
Mol Med Rep ; 23(2)2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33355374

RESUMO

Patients with antiphospholipid syndrome have been identified to have higher incidence rates of atherosclerosis (AS) due to the elevated levels of anti­ß2­glycoprotein I (ß2GPI) antibody (Ab). Our previous studies revealed that the anti­ß2GPI Ab formed a stable oxidized low­density lipoprotein (oxLDL)/ß2GPI/anti­ß2GPI Ab complex, which accelerated AS development by promoting the accumulation of lipids in macrophages and vascular smooth muscle cell. However, the effects of the complex on endothelial cells, which drive the initiation and development of AS, remain unknown. Thus, the present study aimed to determine the proinflammatory roles of the oxLDL/ß2GPI/anti­ß2GPI Ab complex in human umbilical vein endothelial cells (HUVECs) in an attempt to determine the underlying mechanism. Reverse transcription­quantitative PCR, enzymy­linked immunosorbent assay, western blotting and immunofluorescence staining were performed to detect the expressions of inflammation related factors and adhesion molecules. Monocyte­binding assay was used to investigate the effects of oxLDL/ß2GPI/anti­ß2GPI Ab complex on monocyte adhesion to endothelial cells. The results demonstrated that the oxLDL/ß2GPI/anti­ß2GPI Ab complex upregulated the expression of Toll­like receptor (TLR)4 and the levels of NF­κB phosphorylation in HUVECs, and subsequently enhanced the expression levels of inflammatory cytokines, including TNF­α, IL­1ß and IL­6, as well as those of adhesion molecules, such as intercellular adhesion molecule 1 and vascular adhesion molecule 1. In addition, the complex facilitated the recruitment of monocytes by promoting the secretion of monocyte chemotactic protein 1 in HUVECs. Notably, the described effects of the oxLDL/ß2GPI/anti­ß2GPI Ab complex in HUVECs were abolished by either TLR4 or NF­κB blockade. In conclusion, these findings suggested that the oxLDL/ß2GPI/anti­ß2GPI Ab complex may induce a hyper­inflammatory state in endothelial cells by promoting the secretion of proinflammatory cytokines and monocyte recruitment, which was discovered to be largely dependent on the TLR4/NK­κB signaling pathway.


Assuntos
Anticorpos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Lipoproteínas LDL , Complexos Multiproteicos , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/metabolismo , beta 2-Glicoproteína I , Anticorpos/química , Anticorpos/farmacologia , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Inflamação/induzido quimicamente , Inflamação/metabolismo , Inflamação/patologia , Lipoproteínas LDL/química , Lipoproteínas LDL/farmacologia , Monócitos/metabolismo , Monócitos/patologia , Complexos Multiproteicos/química , Complexos Multiproteicos/farmacologia , Células THP-1 , beta 2-Glicoproteína I/química , beta 2-Glicoproteína I/farmacologia
10.
Arch Pharm Res ; 43(7): 744-754, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32715385

RESUMO

Ganoderic Acid A (GA) has many pharmacological effects such as anti-tumor, antibacterial, anti-inflammatory, and immunosuppressive effects. However, the protective effect of GA on liver injury has not been reported. This study aimed to investigate the action of GA on insufficient methionine and choline combined with high-fat diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD) in rats. NAFLD model was established by insufficient methionine and choline combined with high fat feeding to rats. The levels of Acetyl-CoA carboxylase, fatty acid synthase, sterol regulatory element binding protein, liver X receptors, AMP-activated protein kinase, peroxisome proliferator-activated receptor α, PPARg coactivator 1α and NF-κB pathway in the liver were detected by western blot. The results of this study demonstrated that the expression of GA can not only significantly decrease the live weight and liver weight per body weight of HFD mice, but also restore the alanine aminotransferase, aspartate aminotransferase, total bilirubin levels, triglyceride and cholesterol in serum. In addition, the expression of GA increased the levels of high-density lipoprotein cholesterol in serum, ameliorated pathological changes and decreased NAS score of mice's liver. In conclusion, the treatment with GA could improve NAFLD in rats by regulating the levels of signaling events involved in free fatty acid production, lipid oxidation and liver inflammation.


Assuntos
Ácidos Heptanoicos/farmacologia , Inflamação/tratamento farmacológico , Lanosterol/análogos & derivados , Metabolismo dos Lipídeos/efeitos dos fármacos , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Substâncias Protetoras/farmacologia , Administração Oral , Animais , Citocinas/antagonistas & inibidores , Citocinas/sangue , Citocinas/metabolismo , Dieta Hiperlipídica/efeitos adversos , Ácidos Heptanoicos/administração & dosagem , Inflamação/induzido quimicamente , Inflamação/metabolismo , Lanosterol/administração & dosagem , Lanosterol/farmacologia , Masculino , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Hepatopatia Gordurosa não Alcoólica/metabolismo , Substâncias Protetoras/administração & dosagem , Ratos , Ratos Sprague-Dawley
11.
Anal Chem ; 92(9): 6734-6740, 2020 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-32285667

RESUMO

A new photoelectrochemical (PEC) "signal-on" sensing platform based on photoactive material Bi2O3-ZnO and CdS quantum dots (QDs) sensitizer was fabricated for ultrasensitive determination of thrombin by constructing supersandwich nanowires. The CdS/ZnO/Bi2O3 sensitization structure with excellent energy level arrangement remarkably improved photoelectric conversion efficiency because of the efficient separation of the electron-hole. Moreover, the DNA supersandwich nanowire is ingeniously synthesized in one step by simple dislocation hybridization, which could carry a large amount of sensitized material CdS QDs. More importantly, with Exonuclease III (Exo III)-assisted multiple amplification, the proposed "signal-on" platform demonstrated a detection range of 10 fM to 1 µM with the detection limit of 1.41 fM for thrombin. Impressively, the PEC platform can successfully detect human serum samples with good accuracy. Above all, the CdS/ZnO/Bi2O3 sensitization photoelectric biosensing platform by using DNA nanowire in combination with Exo III-multiple amplification opens new sensitized amplification paths for supersensitive biosensing and bioanalysis.


Assuntos
Compostos de Cádmio/química , DNA/química , Técnicas Eletroquímicas , Nanofios/química , Pontos Quânticos/química , Sulfetos/química , Trombina/análise , Humanos , Tamanho da Partícula , Processos Fotoquímicos , Propriedades de Superfície
12.
Analyst ; 145(7): 2805-2810, 2020 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-32103211

RESUMO

In this work, a new kind of dendritically amplified fluorescent signal probe on SiO2 microspheres was controllably fabricated by the terminal deoxynucleotidyl transferase (TdT)-catalyzed incorporation of nucleotides combined with bio-barcode (BBC) amplification for the ultrasensitive detection of Hg2+. A thymine T-Hg2+-T hairpin structure was first formed and further initiated the strand displacement amplification (SDA) reaction, generating a mimic target (MT). MT hybridized with a capture probe 1 (C1) on SiO2 microspheres, and the 3'-hydroxyl (OH) termini of MT initiated TdT-based DNA extension, producing abundant poly-guanine sequences (G1). Then, G1 hybridized with a capture probe 2 (C2) with abundant cytosine (C) species to assemble multiple C2/reporter probe-AuNPs onto the SiO2 microspheres. The reporter DNA further initiated TdT-based extension with a poly-T sequence (T1) to link large numbers of signal probes, which generated a very high fluorescence signal for the ultrasensitive detection of target Hg2+. This TdT-based signal amplification method coupled with SDA exhibits extraordinary sensitivity for Hg2+ assay with a limit down to 1.0 aM. The proposed highly sensitive fluorescence strategy holds great potential for detecting targets in environmental and biological fields.


Assuntos
Mercúrio/análise , Microesferas , Dióxido de Silício/química , Espectrometria de Fluorescência/métodos , DNA/química , DNA/metabolismo , DNA Nucleotidilexotransferase/metabolismo , Corantes Fluorescentes/química , Água Doce/análise , Ouro/química , Mercúrio/química , Nanopartículas Metálicas/química , Técnicas de Amplificação de Ácido Nucleico , Timina/química
13.
J Pharm Biomed Anal ; 172: 86-93, 2019 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-31029803

RESUMO

Dispersive micro-solid-phase extraction (DMSPE) combined with dispersive liquid-liquid microextraction based on the solidification of a floating organic droplet (DLLME-SFO) was successfully developed for extraction and depuration of pantoprazole in rat plasma. The remarkable metal organic framework (MOF), MIL-101(Cr) was used as DMSPE adsorbent. The detection of pantoprazole was performed by convenient HPLC-UV. In the extraction of pantoprazole from plasma samples, small molecule compounds, including the target and abundant impurities, were easily admitted into the porous structure of MIL-101 (Cr) material in DMSPE; while, macromolecular compounds were handily excluded from the adsorbent. Next, the depuration process was achieved by removal of small polar impurities in DLLME-SFO. Influential factors were systematically optimized for ideal enrichment and depuration efficiency. Under the optimal conditions, a satisfactory linearity range from the lower limit of quantification (LLOQ, 100 ng/L) to 10 000 ng/L with the correlation coefficients (r) of 0.9934 was obtained. The LLOQ was 100 ng/mL and the relative recoveries were ≧ 73.2 ±â€¯4.8%. The approving reproducibility, acceptable accuracy, and stability were all within the acceptance limits. This proposed method presented the advantages of environment-friendly, low-cost, recyclable, low impurity, and preferable applicability. It could offer a new idea for the pretreatment and pharmacokinetic study of pantoprazole in rat plasma.


Assuntos
Microextração em Fase Líquida/métodos , Estruturas Metalorgânicas/química , Pantoprazol/sangue , Inibidores da Bomba de Prótons/sangue , Microextração em Fase Sólida/métodos , Animais , Química Farmacêutica , Estudos de Viabilidade , Química Verde/métodos , Masculino , Pantoprazol/isolamento & purificação , Pantoprazol/farmacocinética , Inibidores da Bomba de Prótons/isolamento & purificação , Inibidores da Bomba de Prótons/farmacocinética , Ratos
14.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(12): 1069-1075, 2019 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-31894004

RESUMO

Objective To evaluate the effect of oxidized low-density lipoprotein/ß2-glycoprotein I/anti-ß2 glycoprotein I antibody (oxLDL/ß2GPI/anti-ß2GPI) complex on the calcification and inflammatory cytokine expression in A7r5 rat vascular smooth muscle cells, and to find out the role of Toll-like receptor 4 (TLR4) signal in this process. Methods The A7r5 cells were intervened with oxLDL, oxLDL/ß2GPI complex, oxLDL/anti-ß2GPI complex, ß2GPI/anti-ß2GPI complex, and oxLDL/ß2GPI/anti-ß2GPI complex for different time, with or without TLR4 inhibitor (TAK-242). Alizarin red staining was used to observe the calcification. Real-time quantitative PCR was applied to detect the total mRNA levels of actin-associated protein SM22α (trangelin) and Runt-related transcription factor 2 (RUNX2). The protein levels of SM22α and RUNX2 were measured by Western blotting. Tumor necrosis factor α (TNF-α) was detected by ELISA. Different concentrations of TNF-α were adopted to stimulate A7r5 cells, and the above methods were operated to examine the changes of SM22 and RUNX2. Results When A7r5 cells were incubated by oxLDL/ß2GPI/anti-ß2GPI complex, more calcified nodules were observed, the expression of SM22α was reduced and RUNX2 expression was enhanced at both mRNA and protein levels. And the complex increased the expression of inflammation cytokine TNF-α. TLR4 inhibitor TAK-242 reversed the phenomenon. Different concentrations of TNF-α could reduce mRNA and protein expression of SM22α while raise RUNX2 expression. Conclusion oxLDL/ß2GPI/anti-ß2GPI complex can increase the expression of cytokine TNF-α, and thus quicken calcification procedure in A7r5 cells, along with the change of the traditional contraction phenotype into the osteogenic phenotype. TLR4 receptor participates in this process.


Assuntos
Complexo Antígeno-Anticorpo/farmacologia , Calcinose , Citocinas/metabolismo , Miócitos de Músculo Liso/metabolismo , Animais , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Lipoproteínas LDL/imunologia , Proteínas dos Microfilamentos/metabolismo , Proteínas Musculares/metabolismo , Ratos , Fator de Necrose Tumoral alfa/metabolismo , beta 2-Glicoproteína I/imunologia
15.
J Mol Cell Biol ; 11(5): 421-432, 2019 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-30215728

RESUMO

Paired amphipathic helix protein (SIN3B) is a transcription corepressor for many genes. Here we show a different regulation mechanism of integrin αV gene expression by SIN3B in human hepatocellular carcinoma (HCC). We first observed a close relationship between Integrin αV and SIN3B expressions in HCC patients and tumor cell lines with different metastatic potentials. Overexpression of SIN3B significantly accelerated the cell migration rate of SMMC-7721, but failed when integrin αV expression was silenced. Interestingly, SIN3B stimulated integrin αV subunit promoter activity only in the presence of sulfatide. Importantly, SIN3B was identified in the complex with sulfatide by mass spectrometry. Fat blot assay indicated that SIN3B specifically interacted with sulfatide. Molecular modeling suggested that sulfatide induced the conformational change of SIN3B from compacted α-helices to a relaxed ß-sheet in PAH2 domain. The data of immunoprecipitation and ChIP assay indicated that altered SIN3B lost the binding affinity with MAD1 and HDAC2, which reduced the recruitment of HDAC2 on integrin αV gene promoter and prevented the deacetylation of the histone 3. In conclusion, this study demonstrated that SIN3B promoted the transcriptional activation of the integrin αV subunit gene promoter by reducing interaction with HDAC2.


Assuntos
Carcinoma Hepatocelular/patologia , Integrina alfaV/metabolismo , Neoplasias Hepáticas/patologia , Proteínas Repressoras/metabolismo , Acetilação , Carcinoma Hepatocelular/metabolismo , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Histona Desacetilase 2/química , Histona Desacetilase 2/metabolismo , Histonas/metabolismo , Humanos , Integrina alfaV/genética , Neoplasias Hepáticas/metabolismo , Simulação de Dinâmica Molecular , Regiões Promotoras Genéticas , Ligação Proteica/efeitos dos fármacos , Estrutura Secundária de Proteína , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Proteínas Repressoras/química , Proteínas Repressoras/genética , Sulfoglicoesfingolipídeos/farmacologia , Transcrição Genética/efeitos dos fármacos
16.
Am J Hematol ; 93(6): 803-809, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29603764

RESUMO

Polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes (POEMS) syndrome is a rare plasma dyscrasia without standard treatment. This phase II prospective trial evaluates the safety and response of 12 cycles of low dose lenalidomide (10 mg) plus dexamethasone (Rdex) in patients with newly diagnosed POEMS syndrome. Forty-one patients (28 men) were enrolled and the median age at diagnosis was 49 years (range, 21-70 years). Twenty-one patients (46%) achieved complete hematologic response and the neurologic response rate was 95%. The median serum vascular endothelial growth factor (VEGF) declined from 5155 pg/mL (range, 534-14 328 pg/mL) to 832 pg/mL (95-6254 pg/mL) after therapy. The overall VEGF response rate was 83%, and the median time to response was 2 months, with a mean VEGF reduction of 43% at the first month. In terms of clinical response, Rdex substantially relieved extravascular volume overload, organomegaly, and pulmonary hypertension. No treatment-related deaths occurred and no patients suffered from lenalidomide-related grade 3 or above adverse events. After a median follow-up of 34 months, median overall survival (OS) and progression-free survival (PFS) were not reached, with an estimated 3-year OS and PFS of 90% and 75%, respectively. In conclusion, Rdex was active with high hematologic, VEGF and organ response rate and well tolerated for patients with newly diagnosed POEMS syndrome. This trial was registered at www.clinicaltrials.gov as #NCT01816620.


Assuntos
Dexametasona/administração & dosagem , Lenalidomida/administração & dosagem , Síndrome POEMS/tratamento farmacológico , Adulto , Idoso , Quimioterapia Combinada/métodos , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Síndrome POEMS/mortalidade , Indução de Remissão , Análise de Sobrevida , Resultado do Tratamento , Fator A de Crescimento do Endotélio Vascular/sangue , Adulto Jovem
17.
Water Res ; 130: 300-311, 2018 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-29306195

RESUMO

The kinetics of fluoride sorption by calcite in the presence of metal ions (Co, Mn, Cd and Ba) have been investigated and modelled using the intra-particle diffusion (IPD), pseudo-second order (PSO), and the Hill 4 and Hill 5 kinetic models. Model comparison using the Akaike Information Criterion (AIC), the Schwarz Bayseian Information Criterion (BIC) and the Bayes Factor allows direct comparison of model results irrespective of the number of model parameters. Information Criterion results indicate "very strong" evidence that the Hill 5 model was the best fitting model for all observed data due to its ability to fit sigmoidal data, with confidence contour analysis showing the model parameters were well constrained by the data. Kinetic results were used to determine the thickness of a calcite permeable reactive barrier required to achieve up to 99.9% fluoride removal at a groundwater flow of 0.1 m.day-1. Fluoride removal half-life (t0.5) values were found to increase in the order Ba ≈ stonedust (a 99% pure natural calcite) < Cd < Co < Mn. A barrier width of 0.97 ± 0.02 m was found to be required for the fluoride/calcite (stonedust) only system when using no factor of safety, whilst in the presence of Mn and Co, the width increased to 2.76 ± 0.28 and 19.83 ± 0.37 m respectively. In comparison, the PSO model predicted a required barrier thickness of ∼46.0, 62.6 & 50.3 m respectively for the fluoride/calcite, Mn and Co systems under the same conditions.


Assuntos
Carbonato de Cálcio/química , Fluoretos/química , Metais Pesados/química , Modelos Teóricos , Poluentes Químicos da Água/química , Purificação da Água/métodos , Adsorção , Teorema de Bayes , Difusão , Água Subterrânea/química , Cinética
19.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 33(7): 865-869, 2017 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-28712390

RESUMO

Objective To study the effects of the complex of oxidized low density lipoprotein/ß2-glycoprotein I/ß2-glycoprotein I antibodies (oxLDL/ß2GPI/ß2GPI-Ab) on the migration of human umbilical vein endothelial cells (HUVECs) and the expression of inflammatory cytokines, and their underlying Toll-like receptor (TLR4) pathway. Methods HUVECs were treated with oxLDL, oxLDL/ß2GPI complex, oxLDL/ß2GPI-Ab complex, oxLDL/ß2GPI/ß2GPI-Ab complex, or lipopolysaccharide (LPS) for a period of time in their corresponding groups. The migration of HUVECs was observed by the wound-healing assay. The mRNA and protein levels of TLR4 in HUVECs were detected by real-time quantitative PCR (qRT-PCR) and Western blotting, respectively. The cells were pretreated with or without TAK-242 (the inhibitor of TLR4) 2 hours before stimulated by corresponding stimulus as described above. Then, the contents of monocyte chemotactic protein-1 (MCP-1), interleukin 1ß (IL-1ß) and IL-6 in cell culture supernatant were determined by ELISA, and their mRNAs were detected by qRT-PCR. Results The oxLDL/ß2GPI/ß2GPI-Ab complex promoted the migration of HUVECs effectively, and increased the expression of TLR4. The oxLDL/ß2GPI/ß2GPI-Ab complex increased the expressions of MCP-1, IL-1ß, and IL-6. TAK-242 could reduce the effects of oxLDL/ß2GPI/ß2GPI-Ab complex. Conclusion The oxLDL/ß2GPI/anti-ß2GPI-Ab complex can promote the migration of HUVECs and the expression of related inflammatory cytokines, and TLR4 may be involved in this process.


Assuntos
Complexo Antígeno-Anticorpo/farmacologia , Aterosclerose/etiologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Lipoproteínas LDL/farmacologia , beta 2-Glicoproteína I/farmacologia , Movimento Celular/efeitos dos fármacos , Citocinas/análise , Citocinas/genética , Células Endoteliais da Veia Umbilical Humana/imunologia , Humanos , Receptor 4 Toll-Like/fisiologia
20.
J Pharm Biomed Anal ; 141: 262-269, 2017 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-28372900

RESUMO

Juglone (JL), as one of the major bioactive components present in the bark of Juglans mandshruica Maxim, exhibits versatile bioactivities, especially anti-cancer activity. To better understand the pharmacokinetic properties of juglone, the protein binding rate of juglone was determined by ultrafiltration method, and the binding affinity and mechanism between JL and human serum albumin (HSA) was investigated in vitro through multi-spectroscopic, thermodynamic, and molecular modeling methods. The binding degree of JL was measured more than 99.0% which suggested that JL had high binding ability to serum albumin. Fluorescence data showed that juglone quench the intrinsic fluorescence of HSA upon forming the JL-HSA nonfluorescent complex at 1:1 stoichiometric proportion, and the complex formation had a high affinity of 104 L·mol-1. Meanwhile, the site marker competitive experiments and the thermodynamic parameters (ΔG=-26.08 kJ·mol-1, ΔH=-16.34 kJ·mol-1, ΔS=32.69 J·mol-1·K-1) indicated that juglone could spontaneously bound to the site I (subdomain IIA) of HAS through hydrophobic and hydrogen bonding interactions. As further revealed by the synchronous fluorescence, three-dimensional fluorescence, Fourier transform infrared (FT-IR) and circular dichroism (CD) spectroscopy, JL could cause conformational and structural alterations of HSA. Additionally, molecular docking was employed to further define the specific binding site and the result was in accordance with the conclusion of experimental analysis. The present work provided reasonable models helping us further understand the pharmacokinetics, pharmacological and toxic effects of JL in vivo and supplied an important insight for the applications of JL in the clinical research.


Assuntos
Naftoquinonas/análise , Sítios de Ligação , Dicroísmo Circular , Humanos , Modelos Moleculares , Simulação de Acoplamento Molecular , Ligação Proteica , Albumina Sérica , Espectrometria de Fluorescência , Espectroscopia de Infravermelho com Transformada de Fourier , Termodinâmica , Ultrafiltração
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