Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 53
Filtrar
Mais filtros

Base de dados
Tipo de documento
Intervalo de ano de publicação
1.
EMBO J ; 41(17): e109205, 2022 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-35880301

RESUMO

Patient-derived organoids and cellular spheroids recapitulate tissue physiology with remarkable fidelity. We investigated how engagement with a reconstituted basement membrane in three dimensions (3D) supports the polarized, stress resilient tissue phenotype of mammary epithelial spheroids. Cells interacting with reconstituted basement membrane in 3D had reduced levels of total and actin-associated filamin and decreased cortical actin tension that increased plasma membrane protrusions to promote negative plasma membrane curvature and plasma membrane protein associations linked to protein secretion. By contrast, cells engaging a reconstituted basement membrane in 2D had high cortical actin tension that forced filamin unfolding and endoplasmic reticulum (ER) associations. Enhanced filamin-ER interactions increased levels of PKR-like ER kinase effectors and ER-plasma membrane contact sites that compromised calcium homeostasis and diminished cell viability. Consequently, cells with decreased cortical actin tension had reduced ER stress and survived better. Consistently, cortical actin tension in cellular spheroids regulated polarized basement membrane membrane deposition and sensitivity to exogenous stress. The findings implicate cortical actin tension-mediated filamin unfolding in ER function and underscore the importance of tissue mechanics in organoid homeostasis.


Assuntos
Actinas , Retículo Endoplasmático , Actinas/metabolismo , Retículo Endoplasmático/metabolismo , Estresse do Retículo Endoplasmático , Células Epiteliais/metabolismo , Filaminas/metabolismo , Fenótipo
2.
Proc Natl Acad Sci U S A ; 120(1): e2209260120, 2023 01 03.
Artigo em Inglês | MEDLINE | ID: mdl-36574668

RESUMO

Nanoparticles (NPs) are confronted with limited and disappointing delivery efficiency in tumors clinically. The tumor extracellular matrix (ECM), whose physical traits have recently been recognized as new hallmarks of cancer, forms a main steric obstacle for NP diffusion, yet the role of tumor ECM physical traits in NP diffusion remains largely unexplored. Here, we characterized the physical properties of clinical gastric tumor samples and observed limited distribution of NPs in decellularized tumor tissues. We also performed molecular dynamics simulations and in vitro hydrogel experiments through single-particle tracking to investigate the diffusion mechanism of NPs and understand the influence of tumor ECM physical properties on NP diffusion both individually and collectively. Furthermore, we developed an estimation matrix model with evaluation scores of NP diffusion efficiency through comprehensive analyses of the data. Thus, beyond finding that loose and soft ECM with aligned structure contribute to efficient diffusion, we now have a systemic model to predict NP diffusion efficiency based on ECM physical traits and provide critical guidance for personalized tumor diagnosis and treatment.


Assuntos
Nanopartículas , Neoplasias , Microambiente Tumoral , Humanos , Difusão , Matriz Extracelular/patologia , Nanopartículas/química , Neoplasias/patologia
3.
Proc Natl Acad Sci U S A ; 118(44)2021 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-34716269

RESUMO

Cells cooperate as groups to achieve structure and function at the tissue level, during which specific material characteristics emerge. Analogous to phase transitions in classical physics, transformations in the material characteristics of multicellular assemblies are essential for a variety of vital processes including morphogenesis, wound healing, and cancer. In this work, we develop configurational fingerprints of particulate and multicellular assemblies and extract volumetric and shear order parameters based on this fingerprint to quantify the system disorder. Theoretically, these two parameters form a complete and unique pair of signatures for the structural disorder of a multicellular system. The evolution of these two order parameters offers a robust and experimentally accessible way to map the phase transitions in expanding cell monolayers and during embryogenesis and invasion of epithelial spheroids.


Assuntos
Fenômenos Biofísicos/fisiologia , Processamento de Imagem Assistida por Computador/métodos , Especificidade de Órgãos/fisiologia , Transição de Fase , Animais , Ciclo Celular , Movimento Celular , Proliferação de Células , Células Epiteliais/citologia , Humanos , Morfogênese , Neoplasias , Esferoides Celulares/citologia , Cicatrização
4.
BMC Med ; 21(1): 327, 2023 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-37635247

RESUMO

BACKGROUND: CD133 is considered a marker for cancer stem cells (CSCs) in several types of tumours, including hepatocellular carcinoma (HCC). Chimeric antigen receptor-specific T (CAR-T) cells targeting CD133-positive CSCs have emerged as a tool for the clinical treatment of HCC, but immunogenicity, the high cost of clinical-grade recombinant viral vectors and potential insertional mutagenesis limit their clinical application. METHODS: CD133-specific CAR-T cells secreting PD-1 blocking scFv (CD133 CAR-T and PD-1 s cells) were constructed using a sleeping beauty transposon system from minicircle technology, and the antitumour efficacy of CD133 CAR-T and PD-1 s cells was analysed in vitro and in vivo. RESULTS: A univariate analysis showed that CD133 expression in male patients at the late stage (II and III) was significantly associated with worse progression-free survival (PFS) (P = 0.0057) and overall survival (OS) (P = 0.015), and a multivariate analysis showed a trend toward worse OS (P = 0.041). Male patients with advanced HCC exhibited an approximately 20-fold higher PD-L1 combined positive score (CPS) compared with those with HCC at an early stage. We successfully generated CD133 CAR-T and PD-1 s cells that could secrete PD-1 blocking scFv based on a sleeping beauty system involving minicircle vectors. CD133 CAR-T and PD-1 s cells exhibited significant antitumour activity against HCC in vitro and in xenograft mouse models. Thus, CD133 CAR-T and PD-1 s cells may be a therapeutically tractable strategy for targeting CD133-positive CSCs in male patients with advanced HCC. CONCLUSIONS: Our study provides a nonviral strategy for constructing CAR-T cells that could also secrete checkpoint blockade inhibitors based on a Sleeping Beauty system from minicircle vectors and revealed a potential benefit of this strategy for male patients with advanced HCC and high CD133 expression (median immunohistochemistry score > 2.284).


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Receptores de Antígenos Quiméricos , Humanos , Masculino , Animais , Camundongos , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/terapia , Receptor de Morte Celular Programada 1 , Receptores de Antígenos Quiméricos/genética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/terapia , Modelos Animais de Doenças , Linfócitos T
5.
Hepatology ; 73(5): 1717-1735, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33682185

RESUMO

BACKGROUND AND AIMS: Cancer-associated fibroblasts (CAFs) are key players in multicellular, stromal-dependent alterations leading to HCC pathogenesis. However, the intricate crosstalk between CAFs and other components in the tumor microenvironment (TME) remains unclear. This study aimed to investigate the cellular crosstalk among CAFs, tumor cells, and tumor-associated neutrophils (TANs) during different stages of HCC pathogenesis. APPROACH AND RESULTS: In the HCC-TME, CAF-derived cardiotrophin-like cytokine factor 1 (CLCF1) increased chemokine (C-X-C motif) ligand 6 (CXCL6) and TGF-ß secretion in tumor cells, which subsequently promoted tumor cell stemness in an autocrine manner and TAN infiltration and polarization in a paracrine manner. Moreover, CXCL6 and TGF-ß secreted by HCC cells activated extracellular signal-regulated kinase (ERK) 1/2 signaling of CAFs to produce more CLCF1, thus forming a positive feedback loop to accelerate HCC progression. Inhibition of ERK1/2 or CLCF1/ciliary neurotrophic factor receptor signaling efficiently impaired CLCF1-mediated crosstalk among CAFs, tumor cells, and TANs both in vitro and in vivo. In clinical samples, up-regulation of the CLCF1-CXCL6/TGF-ß axis exhibited a marked correlation with increased cancer stem cells, "N2"-polarized TANs, tumor stage, and poor prognosis. CONCLUSIONS: This study reveals a cytokine-mediated cellular crosstalk and clinical network involving the CLCF1-CXCL6/TGF-ß axis, which regulates the positive feedback loop among CAFs, tumor stemness, and TANs, HCC progression, and patient prognosis. These results may support the CLCF1 cascade as a potential prognostic biomarker and suggest that selective blockade of CLCF1/ciliary neurotrophic factor receptor or ERK1/2 signaling could provide an effective therapeutic target for patients with HCC.


Assuntos
Fibroblastos Associados a Câncer/patologia , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Fibroblastos Associados a Câncer/metabolismo , Carcinoma Hepatocelular/metabolismo , Quimiocina CXCL6/metabolismo , Citocinas/metabolismo , Progressão da Doença , Feminino , Humanos , Neoplasias Hepáticas/metabolismo , Sistema de Sinalização das MAP Quinases , Masculino , Pessoa de Meia-Idade , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo , Microambiente Tumoral
6.
Sensors (Basel) ; 21(11)2021 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-34072991

RESUMO

Traditional precision measurement adopts discrete artificial static observation, which cannot meet the demands of the dynamic, continuous, fine and high-precision holographic measurement of large-scale infrastructure construction and complex operation and maintenance management. Due to its advantages of fast, accurate and convenient measurement, mobile laser scanning technology is becoming a popular technology in the maintenance and measurement of infrastructure construction such as tunnels. However, in some environments without satellite signals, such as indoor areas and underground spaces, it is difficult to obtain 3D data by means of mobile measurement technology. This paper proposes a method to restore the linear of the point cloud obtained by mobile laser scanning based on the measured track center line. In this paper, the measured track position is interpolated with a cubic spline to calculate the translations, and the rotation parameters are calculated by combining the simulation design data. The point cloud of the cross-section of the tunnel under the local coordinate system is converted to the absolute coordinate system to calculate the tunnel line. In addition, the method is verified by experiments combined with the subway tunnel data, and the overall point error can be controlled to within 0.1 m. The average deviation in the horizontal direction is 0.0551 m, and that in the vertical direction is 0.0274 m. Compared with the previous methods, this method can effectively avoid the obvious deformation of the tunnel and the sharp increase in the error, and can process the tunnel point cloud data more accurately and quickly. It also provides better data support for subsequent tunnel analysis such as 3D display, completion survey, systematic hazard management and so on.

7.
Br J Cancer ; 123(10): 1521-1534, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32801345

RESUMO

BACKGROUND: High probability of metastasis limited the long-term survival of patients with hepatocellular carcinoma (HCC). Our previous study revealed that Galectin-3 was closely associated with poor prognosis in HCC patients. METHODS: The effects of Galectin-3 on tumour metastasis were investigated in vitro and in vivo, and the underlying biological and molecular mechanisms involved in this process were evaluated. RESULTS: Galectin-3 showed a close correlation with vascular invasion and poor survival in a large-scale study in HCC patients from multiple sets. Galectin-3 was significantly involved in diverse metastasis-related processes in HCC cells, such as angiogenesis and epithelial-to-mesenchymal transition (EMT). Mechanistically, Galectin-3 activated the PI3K-Akt-GSK-3ß-ß-catenin signalling cascade; the ß-catenin/TCF4 transcriptional complex directly targeted IGFBP3 and vimentin to regulate angiogenesis and EMT, respectively. In animal models, Galectin-3 enhanced the tumorigenesis and metastasis of HCC cells via ß-catenin signalling. Moreover, molecular deletion of Galectin-3-ß-catenin signalling synergistically improved the antitumour effect of sorafenib. CONCLUSIONS: The Galectin-3-ß-catenin-IGFBP3/vimentin signalling cascade was determined as a central mechanism controlling HCC metastasis, providing possible biomarkers for predicating vascular metastasis and sorafenib resistance, as well as potential therapeutic targets for the treatment of HCC patients.


Assuntos
Carcinoma Hepatocelular/patologia , Galectina 3/fisiologia , Neoplasias Hepáticas/patologia , beta Catenina/genética , Animais , Carcinogênese/genética , Carcinogênese/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/mortalidade , Adesão Celular/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Progressão da Doença , Transição Epitelial-Mesenquimal/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/mortalidade , Camundongos Endogâmicos BALB C , Camundongos Nus , Metástase Neoplásica , Neoplasias de Tecido Vascular/genética , Neoplasias de Tecido Vascular/mortalidade , Neoplasias de Tecido Vascular/secundário , Análise de Sobrevida , Análise Serial de Tecidos , Via de Sinalização Wnt/genética , beta Catenina/metabolismo
8.
Proc Natl Acad Sci U S A ; 114(36): 9529-9534, 2017 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-28827333

RESUMO

Active transport in the cytoplasm plays critical roles in living cell physiology. However, the mechanical resistance that intracellular compartments experience, which is governed by the cytoplasmic material property, remains elusive, especially its dependence on size and speed. Here we use optical tweezers to drag a bead in the cytoplasm and directly probe the mechanical resistance with varying size a and speed V We introduce a method, combining the direct measurement and a simple scaling analysis, to reveal different origins of the size- and speed-dependent resistance in living mammalian cytoplasm. We show that the cytoplasm exhibits size-independent viscoelasticity as long as the effective strain rate V/a is maintained in a relatively low range (0.1 s-1 < V/a < 2 s-1) and exhibits size-dependent poroelasticity at a high effective strain rate regime (5 s-1 < V/a < 80 s-1). Moreover, the cytoplasmic modulus is found to be positively correlated with only V/a in the viscoelastic regime but also increases with the bead size at a constant V/a in the poroelastic regime. Based on our measurements, we obtain a full-scale state diagram of the living mammalian cytoplasm, which shows that the cytoplasm changes from a viscous fluid to an elastic solid, as well as from compressible material to incompressible material, with increases in the values of two dimensionless parameters, respectively. This state diagram is useful to understand the underlying mechanical nature of the cytoplasm in a variety of cellular processes over a broad range of speed and size scales.


Assuntos
Citoplasma/química , Citoplasma/fisiologia , Trifosfato de Adenosina/metabolismo , Animais , Fenômenos Biomecânicos , Citoplasma/efeitos dos fármacos , Citoesqueleto/química , Elasticidade , Células Epiteliais/citologia , Células HeLa/citologia , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Humanos , Rim/citologia , Miosina Tipo II/antagonistas & inibidores , Miosina Tipo II/metabolismo , Pinças Ópticas , Ratos , Viscosidade
9.
Proc Natl Acad Sci U S A ; 114(41): E8618-E8627, 2017 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-28973866

RESUMO

Cells alter their mechanical properties in response to their local microenvironment; this plays a role in determining cell function and can even influence stem cell fate. Here, we identify a robust and unified relationship between cell stiffness and cell volume. As a cell spreads on a substrate, its volume decreases, while its stiffness concomitantly increases. We find that both cortical and cytoplasmic cell stiffness scale with volume for numerous perturbations, including varying substrate stiffness, cell spread area, and external osmotic pressure. The reduction of cell volume is a result of water efflux, which leads to a corresponding increase in intracellular molecular crowding. Furthermore, we find that changes in cell volume, and hence stiffness, alter stem-cell differentiation, regardless of the method by which these are induced. These observations reveal a surprising, previously unidentified relationship between cell stiffness and cell volume that strongly influences cell biology.


Assuntos
Diferenciação Celular , Fenômenos Fisiológicos Celulares , Tamanho Celular , Células-Tronco Mesenquimais/fisiologia , Água/metabolismo , Animais , Linhagem da Célula , Células Cultivadas , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos BALB C
10.
Sensors (Basel) ; 20(18)2020 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-32967174

RESUMO

Subway structure safety detection is an important method to ensure the safe operation of trains. Efficient, high-precision, and automatic tunnel clearance detection is the key to ensure safe operations. This study introduces a mobile tunnel scanning system that integrates a scanner, an inertial measurement unit (IMU), and a rail car. Global Navigation Satellite System (GNSS) time and system hardware calibration are used to synchronize time and space information of the system; the attitude and speed are corrected using the control points from the tunnel to improve the accuracy of absolute positioning. The section coordinate system is converted using the control points and system calibration parameters to complete the tunnel clearance inspection, and the distance between the nearest point of the section and the clear height of the vault is given. Taking Fengxi Road's Bashan tunnel section of Chongqing Metro Line 5 as an example, the overall system accuracy was tested. The accuracy of chord line measurements was within 1 mm, the internal coincidence accuracy of repeated measurements of the vault clear height was 1.1 mm, the internal coincidence accuracy of repeated measurements of the closest gauge point was 4.8 mm, and the system calibration accuracy was approximately 2 mm. Compared with the existing scheme, the system combines absolute measurement and relative measurement mode to judge the structural safety of tunnel section from multiple angles, high precision, and high efficiency.

11.
Biol Reprod ; 99(6): 1205-1215, 2018 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-30010724

RESUMO

A variety of mechanisms are involved in sex determination in vertebrates. The orange-spotted grouper (Epinephelus coioides), a teleost fish, functions first as females and later as a male and is an ideal model to investigate the regulation of sexual fate. Here, we report female-to-male sex reversal in juvenile orange-spotted groupers caused by overexpressing anti-Müllerian hormone (Amh). Tissue distribution analyses showed that amh and amhrII primarily expressed in the gonad, and expression level in the testis was much higher than that in the ovary. In gonads, the expression of amh was located in the Sertoli cells around spermatogonia of the testis and in the zona pellucida of the mature ovary, and the expression of amhrII was located in the Sertoli cells of the testis and in the oocytes of the ovary. Decrease in female-related genes and serum 17ß-estradiol level, increase in male-related genes and serum 11-ketotestosterone, ovarian regression, and spermatogonia proliferation were observed during plasmid feeding experiment. These results illustrate that amh overexpression plasmid feeding can induce a female-to-male transition in grouper.


Assuntos
Hormônio Antimülleriano/metabolismo , Perciformes/fisiologia , Processos de Determinação Sexual/fisiologia , Animais , Hormônio Antimülleriano/genética , Estradiol/sangue , Feminino , Regulação da Expressão Gênica , Masculino , Ovário/metabolismo , Plasmídeos , Receptores de Peptídeos/genética , Receptores de Peptídeos/metabolismo , Receptores de Fatores de Crescimento Transformadores beta/genética , Receptores de Fatores de Crescimento Transformadores beta/metabolismo , Diferenciação Sexual , Testículo/metabolismo , Testosterona/análogos & derivados , Testosterona/sangue , Transcriptoma
12.
Gen Comp Endocrinol ; 267: 90-97, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-29913168

RESUMO

Luteinizing hormone receptor (LHR) plays a critical role in reproduction by mediating LH signaling in the gonad. In this study, we cloned a novel lhr gene from the orange-spotted grouper, named glhr2. The cloned complete open reading frame sequence of glhr2 was 2082 bp in length, encoding a protein of 693 amino acids, sharing approximately 50% amino acid identity with glhr1. glhr1 and glhr2 were primarily expressed in gonad, brain and hypothalamus with low expression in other tissues such as gill, spleen, etc. The expressions of both glhr1 and glhr2 increased during vitellogenesis, while decreased during natural female to male sex change. The two gLHRs both could be activated by equine LH or human chorionic gonadotropin, but not by human follicle stimulating hormone. Both gLHR1 and gLHR2 activation stimulated the expression of cAMP response element driven reporter gene in a dose-dependent manner, while gLHR2 but not gLHR1 also activated serum response element driven reporter gene expression. This was the first study to demonstrate that two active LHRs exist in fish with possible different functional roles.


Assuntos
Perciformes/genética , Receptores do LH/genética , Receptores do LH/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Gonadotropina Coriônica/farmacologia , Clonagem Molecular , DNA Complementar/genética , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Gônadas/citologia , Gônadas/efeitos dos fármacos , Gônadas/metabolismo , Cavalos , Humanos , Ligantes , Luciferases/metabolismo , Masculino , Fases de Leitura Aberta/genética , Perciformes/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores do LH/química
13.
J Virol ; 89(15): 7722-34, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25972549

RESUMO

UNLABELLED: Human noroviruses (HuNoVs) are positive-sense RNA viruses that can cause severe, highly infectious gastroenteritis. HuNoV outbreaks are frequently associated with recombination between circulating strains. Strain genotyping and phylogenetic analyses show that noroviruses often recombine in a highly conserved region near the junction of the viral polyprotein (open reading frame 1 [ORF1]) and capsid (ORF2) genes and occasionally within the RNA-dependent RNA polymerase (RdRP) gene. Although genotyping methods are useful for tracking changes in circulating viral populations, they report only the dominant recombinant strains and do not elucidate the frequency or range of recombination events. Furthermore, the relatively low frequency of recombination in RNA viruses has limited studies to cell culture or in vitro systems, which do not reflect the complexities and selective pressures present in an infected organism. Using two murine norovirus (MNV) strains to model coinfection, we developed a microfluidic platform to amplify, detect, and recover individual recombinants following in vitro and in vivo coinfection. One-step reverse transcriptase PCR (RT-PCR) was performed in picoliter drops with primers that identified the wild-type and recombinant progenies and scanned for recombination breakpoints at ∼1-kb intervals. We detected recombination between MNV strains at multiple loci spanning the viral protease, RdRP, and capsid ORFs and isolated individual recombinant RNA genomes that were present at a frequency of 1/300,000 or higher. This study is the first to examine norovirus recombination following coinfection of an animal and suggests that the exchange of RNA among viral genomes in an infected host occurs in multiple locations and is an important driver of genetic diversity. IMPORTANCE: RNA viruses increase diversity and escape host immune barriers by genomic recombination. Studies using a number of viral systems indicate that recombination occurs via template switching by the virus-encoded RNA-dependent RNA polymerase (RdRP). However, factors that govern the frequency and positions of recombination in an infected organism remain largely unknown. This work leverages advances in the applied physics of drop-based microfluidics to isolate and sequence rare recombinants arising from the coinfection of mice with two distinct strains of murine norovirus. This study is the first to detect and analyze norovirus recombination in an animal model.


Assuntos
Infecções por Caliciviridae/virologia , Norovirus/genética , Norovirus/isolamento & purificação , Recombinação Genética , Animais , Variação Genética , Genótipo , Humanos , Camundongos , Microfluídica , Dados de Sequência Molecular , Norovirus/classificação , Filogenia
14.
Zhongguo Zhong Yao Za Zhi ; 40(22): 4446-51, 2015 Nov.
Artigo em Zh | MEDLINE | ID: mdl-27097422

RESUMO

To investigate the practicability of establishing zebrafish lipid-lowering drug screening model and the effect of berberine (BBR) on hyperlipidemic zebrafish. Three-month-old zebrafishes were fed with 4% cholesterol for 0, 2, 4, 8, 14, 20, 25, 30 days, and the level of total cholesterol in serum was measured. Zebrafish were randomly divided into four groups: the control group, the high cholesterol diet group, the 0.01% simvastatin-treated group, the 0.1% berberine-treated group and the 0.2% berberine-treated group. The levels of total cholesterol (TC), triglyceride (TC), low density lipoprotein cholesterol (LDL-c) and high-density lipoprotein cholesterol (HDL-c) in serum were measured; the expression of hepatic HMGCR, LDLR and CYP7A1a mRNA expressions were detected by real time PCR. Oil red O staining was performed to observe the changes in fat content in the liver. According to the result, the level of serum TC in the 4% cholesterol diet group significantly was higher than that of the normal control group in a time-dependent manner and reached a stable level at the 20th day. The BBR group showed significant decreases in the levels of TC, TG and LDL-c, HMGCR mRNA expression and fat content and increases in LDLR and CYP7A1a mRNA. The hyperlipidemia zebrafish model was successfully established by feeding with 4% cholesterol for 20 days. The findings lay a foundation for further screenings on lipid-lowering drugs.


Assuntos
Berberina/administração & dosagem , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/administração & dosagem , Hiperlipidemias/tratamento farmacológico , Hipolipemiantes/administração & dosagem , Peixe-Zebra/metabolismo , Animais , Colesterol/metabolismo , Feminino , Humanos , Hiperlipidemias/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Triglicerídeos/metabolismo
15.
Biosci Biotechnol Biochem ; 78(3): 438-47, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25036830

RESUMO

Protein kinases are key regulators of cell function that constitute one of the largest and most functionally diverse gene families. We developed a novel assay system, based on the bimolecular fluorescence complementation (BiFC) technique in Escherichia coli, for detecting transient interactions such as those between kinases and their substrates. This system detected the interaction between OsMEK1 and its direct target OsMAP1. By contrast, BiFC fluorescence was not observed when OsMAP2 or OsMAP3, which are not substrates of OsMEK1, were used as prey proteins. We also screened for interacting proteins of calcium-dependent protein kinase 8 (OsCPK8), a regulator of plant immune responses, and identified three proteins as interacting molecules of OsCPK8. The interaction between OsCPK8 and two of these proteins (ARF-GEF and peptidyl prolyl isomerase) was confirmed in rice cells by means of BiFC technology. These results indicate that our new assay system has the potential to screen for protein kinase target molecules.


Assuntos
Oryza/metabolismo , Proteínas de Plantas/metabolismo , Proteínas Quinases/metabolismo , Escherichia coli , Microscopia de Fluorescência , Oryza/genética , Proteínas de Plantas/genética , Proteínas de Plantas/isolamento & purificação , Mapas de Interação de Proteínas/genética , Proteínas Quinases/genética
16.
J Nanosci Nanotechnol ; 13(2): 1208-11, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23646604

RESUMO

Osteosarcoma and chondrosarcoma are malignant bone tumors, and they significantly affect the life quality of patients including children and adults. The main treatment method is surgical amputation of the malignant lesion, despite that recurrence often occurs. Recently, it has been observed that TiO2 NPs killed HeLa cells effectively via photocatalysis in vitro, which indicates titanium dioxide (TiO2) nanoparticles (NPs) might be used to reduce the recurrence of osteosarcoma and chondrosarcoma by inducing cytotoxicity to bone tumor cells. In this study, we investigated the potential effects of TiO2 NPs in two cancer cell lines in vitro: U-2 OS (osteosarcoma) and SW 1353 (chondrosarcoma). We assessed cell viability, the levels of reactive oxygen species (ROS) and glutathione (GSH) after exposure to TiO2 NPs at different concentrations (0.1-100 microg/ml) for varying exposure periods (12-48 hours). Compared to the NP-free control, TiO2 NPs induced cell death in a dosage-dependent and time-dependent manner. The median inhibitory concentration (IC50) of TiO2 NPs at 24 hours was 211.3 +/- 15.2 microg/ml and 5408.8 +/- 45.9 microg/ml for SW 1353 and U-2 OS cell lines, respectively. TiO2 NPs concentrations above 1 microg/ml were more efficient to reduce the cell viability of SW 1353 than U-2 OS of NPs at all exposure times. The increased ROS and reduced GSH levels indicated that TiO2 NPs killed cancer cells through oxidative stress. These results suggested that the TiO2 NPs can be potentially used to minimize/prevent the recurrence of osteosarcoma and chondrosarcoma.


Assuntos
Condrossarcoma/prevenção & controle , Nanopartículas Metálicas , Recidiva Local de Neoplasia , Osteossarcoma/prevenção & controle , Titânio/química , Condrossarcoma/metabolismo , Condrossarcoma/patologia , Glutationa/metabolismo , Humanos , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Espécies Reativas de Oxigênio/metabolismo
17.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi ; 30(4): 782-8, 2013 Aug.
Artigo em Zh | MEDLINE | ID: mdl-24059056

RESUMO

To develop standard in vitro chondrosarcoma models, we synthesized three hydrogels (i. e., PDMAAm, PNaAMPS and PMETAC) and investigated the influence of Young's modulus, swelling ratio and electric charges on the behavior of chondrosarcoma cells seeded on the hydrogels, including morphology, adhesion and aggregation. Results showed that the morphology of chondrosarcoma cells at 6h was dependent on the charges of hydrogels; cells present spindle-shaped and round-shaped morphology on negative charged and neutral hydrogel, respectively, while no cells spreaded on positive charged hydrogel. Chondrosarcoma cells formed aggregates on neutral PDMAAm after further culture. The hydrogels can be synthesized easily and has the characteristics of ease at use with defined components, which holds great potential for developing standard chondrosarcoma models in vitro.


Assuntos
Neoplasias Ósseas/patologia , Proliferação de Células/efeitos dos fármacos , Condrossarcoma/patologia , Hidrogéis/farmacologia , Linhagem Celular Tumoral , Humanos , Hidrogéis/química , Metacrilatos/farmacologia , Nylons/farmacologia , Eletricidade Estática
18.
Medicine (Baltimore) ; 102(40): e35338, 2023 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-37800786

RESUMO

The monocyte-to-high-density lipoprotein cholesterol (HDL) ratio (MHR) is accepted as a novel marker for demonstrating inflammation. We investigated whether the monocyte-to-HDL ratio is related to the 90-day clinical prognosis of acute ischemic stroke (AIS) after intravenous thrombolysis (IVT). Patients with AIS treated with alteplase IVT were included consecutively, and clinical information and laboratory data were collected. The 90-day prognosis of all patients was determined with a clinical assessment using the modified Rankin Scale (mRS). The optimal cutoff values for patients were evaluated by the receiver operating characteristic (ROC) curve method. Then, a multivariate logistic regression model was used to evaluate the risk factors for poor prognosis of IVT in AIS. We retrospectively enrolled 192 patients who were diagnosed with AIS and received IVT between February 2020 and July 2022, with final follow-up on September 30, 2022. The patients in the poor prognosis group had significantly higher monocyte counts, lower HDL levels, and higher MHR values than the good prognosis group. The optimal cutoff value of the MHR for predicting the 3-month outcome of acute pontine infarction was 0.621. Multivariate logistic regression revealed that the MHR (OR = 4.626, 95% CI: 1.156-18.512, P = .030) was strongly associated with poor prognosis in AIS. The MHR is an independent risk factor for the clinical prognosis of AIS patients receiving IVT therapy and shows a certain predictive value.


Assuntos
Isquemia Encefálica , AVC Isquêmico , Acidente Vascular Cerebral , Humanos , Monócitos , HDL-Colesterol , AVC Isquêmico/tratamento farmacológico , Acidente Vascular Cerebral/tratamento farmacológico , Isquemia Encefálica/tratamento farmacológico , Estudos Retrospectivos , Prognóstico , Terapia Trombolítica
19.
Exploration (Beijing) ; 3(5): 20220161, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37933283

RESUMO

Tumor cells may be eliminated by increasing their temperature. This is achieved via photothermal therapy (PTT) by penetrating the tumor tissue with near-infrared light and converting light energy into heat using photothermal agents. Copper sulfide nanoparticles (CuS NPs) are commonly used as PTAs in PTT. In this review, we aimed to discuss the synergism between tumor PTT with CuS NPs and other therapies such as chemotherapy, radiotherapy, dynamic therapies (photodynamic, chemodynamic, and sonodynamic therapy), immunotherapy, gene therapy, gas therapy, and magnetic hyperthermia. Furthermore, we summarized the results obtained with a combination of two treatments and at least two therapies, with PTT as one of the included therapies. Finally, we summarized the benefits and drawbacks of various therapeutic options and state of the art CuS-based PTT and provided future directions for such therapies.

20.
Animals (Basel) ; 13(13)2023 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-37443982

RESUMO

The objective of this study was to investigate the effects of dietary supplementation with different types of potassium and magnesium on the reproductive performance, antioxidant capacity, and immunity of sows. Forty-five Landrace × Yorkshire sows at the late gestation stage (85 d) were randomly assigned to three groups (n = 15). Sows in the control group (CON), potassium chloride and magnesium sulfate group (PM), and potassium-magnesium sulfate group (PMS) were fed with a basal diet, a basal diet supplemented with magnesium sulfate (0.20%) and potassium chloride (0.15%), or a basal diet supplemented with potassium-magnesium sulfate (0.45%), respectively. The results showed that dietary supplementation with PMS did not yield significant effects on the reproductive performance compared with the CON group (p > 0.05). However, it significantly elevated the level of insulin-like growth factor 1 (IGF-1) in plasma and immunoglobulin A (IgA) in colostrum (p < 0.05). Furthermore, PMS significantly augmented the activities of catalase (CAT) and superoxide dismutase (SOD) while reducing the levels of malondialdehyde (MDA) in comparison to the CON group (p < 0.05). Compared with the PM group, the PMS group significantly reduced the incidence rate of intrauterine growth restriction (IUGR) (p < 0.05) and significantly decreased the concentration of the proinflammatory cytokine (TNF-α) level in plasma (p < 0.05). These results indicated that dietary supplementation with PMS during late gestation could enhance sows' antioxidant capacity and the IgA level in colostrum. These findings will provide a theoretical reference for the use of magnesium and potassium in sow production to improve sows' health.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA