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1.
Cell Death Dis ; 12(1): 87, 2021 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-33462188

RESUMO

Wilson's disease (WD) is an inherited disorder characterized by excessive accumulation of copper in the body, particularly in the liver and brain. In the central nervous system (CNS), extracellular copper accumulation triggers pathological microglial activation and subsequent neurotoxicity. Growing evidence suggests that levels of inflammatory cytokines are elevated in the brain of murine WD models. However, the mechanisms associated with copper deposition to neuroinflammation have not been completely elucidated. In this study, we investigated how the activation of NLR family pyrin domain containing 3 (NLRP3) inflammasome contributes to copper-mediated neuroinflammation in an animal model of WD. Elevated levels of interleukin-1ß, interleukin-18, interleukin-6, and tumor necrosis factor-α were observed in the sera of WD patients and toxic milk (TX) mice. The protein levels of inflammasome adaptor molecule apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC), cleaved caspase-1, and interleukin-1ß were upregulated in the brain regions of the TX mice. The NLRP3 inflammasome was activated in the TX mice brains. Furthermore, the activation of NLRP3 inflammasome was noted in primary microglia treated with CuCl2, accompanied by the increased levels of cleaved caspase-1, ASC, and interleukin-1ß. Blocking NLRP3 inflammasome activation with siNlrp3 or MCC950 reduced interleukin-1ß and interleukin-18 production, thereby effectively mitigating cognitive decline, locomotor behavior impairment, and neurodegeneration in TX mice. Overall, our study demonstrates the contribution of copper overload-mediated activation of NLRP3 inflammasome to progressive neuropathology in the CNS of a murine model of WD. Therefore, blockade of the NLRP3 inflammasome activation could be a potential therapeutic strategy for WD.

2.
Free Radic Biol Med ; 2021 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-33450374

RESUMO

Metastasis is the major cause of death in women with advanced ovarian cancer. Epithelial ovarian cancer cells can dissociate directly from extracellular matrix (ECM) and form spheroids to spread through the peritoneal cavity. Loss of ECM hinders the survival of ECM-detached epithelial cells. It is still largely unknown how ovarian cancer spheroids maintain their viability after loss of ECM. We find that spheroids derived either from ovarian cancer ascites or cell lines are iron-replete. In accordance with iron-replete condition, proteins involved in iron uptake, transport and storage including divalent metal ion transporter 1 (DMT1), transferrin receptor 1 (TFR1), ferritin, poly(rC)-binding proteins 1 and 2 (PCBP1 and 2) and nuclear factor E2-related factor 2 (NRF2) all increase in ovarian cancer spheroids. Genes linking iron homeostasis and lipid metabolism including stearoyl coenzyme A desaturase 1 (SCD1) are up-regulated in ovarian cancer spheroids. The product of SCD1 oleic acid can restore the viability of ovarian cancer spheroids inhibited by deprivation of iron. Extracellular signal-regulated kinase (ERK) activation contributes to autophagy activation in ovarian cancer spheroids. Impairment of autophagy by U0126 or Olaparib results in lysosomal iron accumulation and decrease of the cytosolic labile iron pool, leading to reduction of SCD1, lipid level and cell viability. Combination of U0126 and Olaparib has synergistic cytotoxicity toward ovarian cancer spheroids. Our findings reveal that ovarian cancer spheroids develop efficient iron utilization system to survive. Targeting iron utilization in ovarian cancer spheroids may have the potential to become new treatment strategies for ovarian cancer metastasis.

3.
Food Chem ; 339: 127955, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32919344

RESUMO

Lateral flow assay (LFA), performed with simple devices and short detection time, is popular in field applications. Herein, a novel sandwich type-based LFA was constructed for high sensitivity and selectivity detection of Staphylococcus aureus (S. aureus). Vancomycin-immobilized gold nanoparticles (VAN-Au NPs) were utilized as the first identifier to capture S. aureus and the specificity was guaranteed by the second recognition agent of pig immunoglobulin G (IgG). In addition, gold growth was adopted for signal amplification to further improve the detection sensitivity. S. aureus could be directly assayed by this LFA within the concentration range of 1.0 × 103-1.0 × 108 cfu mL-1 with a detection limit of 1.0 × 103 cfu mL-1. Furthermore, the novel sandwich LFA realized S. aureus detection in food samples with admissible recoveries and established a rapid, simple, cost-effective and sensitive platform, could meet the demand for on-site testing of S. aureus.


Assuntos
Antibacterianos/química , Imunoensaio/métodos , Imunoglobulina G/química , Staphylococcus aureus/isolamento & purificação , Animais , Microbiologia de Alimentos , Sucos de Frutas e Vegetais/microbiologia , Ouro/química , Imunoglobulina G/imunologia , Limite de Detecção , Nanopartículas Metálicas/química , Sistemas Automatizados de Assistência Junto ao Leito , Staphylococcus aureus/imunologia , Suínos , Vancomicina/química , Verduras/microbiologia
4.
Carbohydr Polym ; 251: 117097, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33142635

RESUMO

Inspired by the seashell nacre and seaweed, a novel GO-Ca2+-SA nacre-inspired hybrid mesh was prepared via an interfacial layer-by-layer self-assembly and cross-linking, using graphene oxide (GO) and sodium alginate (SA) as the building blocks and calcium chloride as the coordination agent, respectively. Hybrid mesh was characterized by FTIR, XPS, XRD, SEM and contact angel instrument, showing superhydrophilic and underwater superoleophobic property and low oil adhesion, due to its wrinkle and rough surface, and high hydration ability of GO-Ca-alginate nanohydrogels. The separation efficiencies of various oil-water mixtures were above 99 %, with a highest flux of 119,426 L m-2 h-1. Hybrid mesh showed an orderly layered "brick and mortar" microstructure with many ultrasmall nanoscaled protuberances. Ca2+ ions could chelate with SA to form the "egg-box" structure, and interact with GO nanosheets. Hybrid mesh possessed high salt/acid/alkaline tolerance, abrasion resistance, mechanical property with Young's modulus of 35.8 ± 4.9 GPa, and excellent cycling stability.

5.
J Agric Food Chem ; 69(1): 511-519, 2021 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-33373219

RESUMO

Gold nanoparticles (AuNPs) are the most commonly used signal materials in lateral flow immunoassay (LFIA). However, the assay sensitivity of traditional AuNP-based LFIA is usually limited by the incomplete competition between free target analytes and immobilized antigens for the binding of AuNP-labeled antibodies. To unfreeze this limitation, here, asymmetric Au-SiO2 Janus NPs (about 66 nm) were designed and synthesized. Au-SiO2 Janus NPs can assemble into snowman-like anisotropic structures and combine two different physicochemical properties at their opposite sides, where the AuNP side mainly possesses the antibody conjugating and signal providing functions and the SiO2 side primarily offers the stable function. In virtue of the unique asymmetric nanostructure, only the AuNP side can interact with target analytes by specific antigen-antibody interactions, which could significantly improve the efficiency of competition. Selecting furazolidone as a model analyte, the immunoassay biosensor showed a limit of detection as low as 0.08 ng/mL, 10-fold decreased than that of the AuNPs-LFIA. Moreover, the Au-SiO2 Janus NP lateral flow immunoassay was well applied in chicken, pork, honey, and beef food samples with visual detection limits of 0.8 ng/g, 0.16 ng/g, 0.4 ng/mL, and 0.16 ng/g, respectively. The Au-SiO2 Janus NPs possess the advantages of both materials, which will broaden their applications as a potential alternative in the rapid and sensitive detection of antibiotic residues.

6.
Am J Cancer Res ; 10(11): 3947-3972, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33294279

RESUMO

The AT-rich Interactive Domain 1A (ARID1A) is one of the most frequently mutated genes in gastric cancer. Here, we found that genetic variants in noncoding regions of ARID1A associated with altered protein levels by target sequencing. Notably, tumors with ARID1A variants in the 3'untranslated region (3'UTR) exhibited remarkably increased heterogeneity of ARID1A protein. In general, genetic variants and protein deficiency of ARID1A in tumors were associated with a better survival. Strikingly, altered patterns and heterogeneity of ARID1A protein expression were observed in peritumor tissues and carried significant implications in defining tumor immune contexture by multiplex immunohistochemistry. By analyzing the spatial distribution of TILs, we showed that reduced ARID1A protein levels in both tumor and peritumor tissues were significantly correlated with increased density and proximity of TILs to tumor cells. In contrast, high heterogeneity of ARID1A expression was associated with increased TIL density, but reduced proximity of TILs to tumor cells. Collectively, our study characterized ARID1A genetic alterations and its protein expression patterns in EOGC, demonstrating new strategies for clinically assessing its molecular impact on tumor onset and progression, tumor immune response, and patient survival.

7.
J Agric Food Chem ; 68(52): 15509-15515, 2020 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-33331779

RESUMO

An innovative lateral flow competition immunoassay (LFCIA) for detecting clenbuterol (CL) was developed by employing the advantages of the coomassie brilliant blue (CBB) staining method. An antibody stained by CBB was used both as a recognition reagent and as a chromogenic probe, enabling the simple but sensitive LFCIA of CL. The CBB-based LFCIA exhibited sensitivity for CL with a detection limit of 2 ng mL-1. Furthermore, this strategy was preliminarily verified by screening for CL in milk, pork tenderloin, and swine liver with recoveries ranging from 81 to 102%. Compared with conventional LFCIAs, the use of CBB as a signal label not only avoided the complicated material synthesis and surface modification process but also simplified the cross-linking with antibodies, meanwhile reducing the steric hindrance and increasing the possibility of immune recognition reactions, which was propitious for the effective utilization of antibodies. Taking advantages of the simplicity, rapidity, and cost-effectiveness, the CBB-based LFCIA may have potential for on-demand monitoring of general harmful small molecules by changing the kind of the staining antibody.

8.
J Biochem Mol Toxicol ; : e22672, 2020 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-33270355

RESUMO

Brahma-related gene 1 (Brg-1) is perceived as a cytoprotective protein due to its role in alleviating oxidative stress and apoptosis. Our study aimed to explore the role and mechanism of Brg-1 in high glucose (HG)-stimulated podocytes. The HG exposure downregulated Brg-1 and inactivated the protein kinase B (Akt) pathway in podocytes. Restoration of Brg-1 inhibited HG-induced viability reduction of podocytes. The HG-induced increase of reactive oxygen species and malondialdehyde levels and decrease of superoxide dismutase activity in podocytes were reversed by the Brg-1 overexpression. The Brg-1 overexpression terminated the HG-induced production of fibronectin, collagen IV, transforming growth factor-ß1, and connective tissue growth factor. In addition, the Brg-1 overexpression activated Akt-dependent nuclear factor E2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling in HG-stimulated podocytes. However, inhibition of the Akt pathway or Nrf2 silencing counteracted the protective effects of Brg-1 in HG-stimulated podocytes. In conclusion, the Brg-1 overexpression suppressed HG-induced oxidative stress and extracellular matrix accumulation by activation of Akt-dependent Nrf2/ARE signaling in podocytes.

9.
J Ethnopharmacol ; 269: 113749, 2020 Dec 25.
Artigo em Inglês | MEDLINE | ID: mdl-33359861

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Aralia echinocaulis has been used in traditional medicines in China and exhibits good effects on rheumatoid arthritis (RA). AIM OF THE STUDY: Aralia echinocaulis is rich in polysaccharides and glycosides. This study aims to explore the effect of total polysaccharide and glycoside (TPG) from A. echinocaulis on an RA rat model and the role of alterations in gut microbes mediated by TPG. MATERIALS AND METHODS: In this study, a collagen-induced arthritis (CIA) rat model was constructed and used to evaluate the effects of TPG in vivo. 16S rRNA sequencing was used to detect the changes in the gut microbiota. A cooccurrence analysis was conducted by calculating Spearman's rank correlations. Microbial functions were predicted using PICRUSt with the KEGG and COG databases. RESULTS: The results showed that TPG from A. echinocaulis could inhibit arthritis, reduce serum IL-1ß and TNF-α levels, and improve synovial pathology in the RA rat model but failed to produce the same results in a pseudoaseptic RA rat model. 16S rRNA sequencing verified that TPG could modulate the gut microbiota community structure of RA rats. The cooccurrence analysis found 19 out of the 50 most abundant genera in a cooccurrence network, of which 16 showed a positive correlation and 3 showed a negative correlation. KEGG pathway and COG function analyses found that TPG-induced alterations in the gut microbiota might be correlated with the circulatory system, excretory system, metabolic diseases, signaling molecules and interactions, coenzyme transport and metabolism, and nucleotide transport and metabolism. CONCLUSIONS: TPG from A. echinocaulis had significant effects on the RA rat model, which are related to the modulation of the gut microbiota. These results are useful to better understanding the mechanisms of TPG in RA.

10.
J Colloid Interface Sci ; 587: 202-213, 2020 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-33360893

RESUMO

Recently, the development of dual functional catalytic membrane for the synergistic degradation and filtration of persistent pollutants has attracted considerable attention in environmental remediation. Herein, novel CoFe alloy and CoFe2O4 nanoparticles encapsulated in N-doped microtube composites (CoFe-NMTs) were firstly fabricated through in-situ pyrolysis of simple-source Prussian blue analogues (PBA). As expected, this unique structure not only inhibited the nanoparticles agglomeration, but also provided a "highway" that accelerated the Co3+/Co2+ and Fe3+/Fe2+ redox cycles. Therefore, CoFe-NMT-800 (0.1 g/L, pyrolyzed at 800 °C) achieved over 90% tetracycline (TC, 30 mg/L, 0.1821 min-1) removal after 30 min at a wide pH (2.55-9.55) by coupling with peroxymonosulfate (PMS, 0.3 g/L), which dramatically outperformed the majority of the reported catalysts (such as Co3O4, CoFe alloy, CoFe2O4 and N-doped carbon nanotubes, etc.). Additionally, CoFe-NMTs-800 also exhibited excellent catalytic activity in the existence of inorganic anions (Cl-, HCO3- and H2PO4-) and natural organic matters (humic acid (HA)). Subsequently, CoFe-NMTs-800 was immobilized into polyvinylidene fluoride (PVDF) membrane as catalytic self-cleaning membrane via applying phase-inversion technology. It was found that CoFe-NMTs-800/PVDF membrane not only maintained high removal efficiency for TC degradation (over 90%) in TC/HA coexistence system, but also effectively eliminated the adverse effect of membrane fouling. Besides, the fabricated membrane also showed desirable reusability and neglectable metal leaching (0.003 mg/L Fe and 0.015 mg/L) with almost constant flux after five cycles. The quenching experiments and electron paramagnetic resonance (EPR) results clearly indicated that sulfate radicals (SO4-), hydroxyl radicals (OH and singlet oxygen (1O2) were responsible for TC degradation and SO4- was a major contributor. Significantly, this work was very meaningful to construct novel catalytic self-cleaning membrane for water purification.

11.
Artigo em Inglês | MEDLINE | ID: mdl-33189323

RESUMO

Currently, carbon-based catalysts integrated with macroporous catalytic membrane have aroused considerable attention for environmental remediation because of its practicability and high efficiency. Herein, nitrogen doped carbon nanotube hybrids (Fe-Co@NC-CNTs) decorated with multiple active species (Fe3Co7/CoFe2O4@Fe/CoNC) were designed through N-molecule assisted pyrolysis of bimetallic (Fe/Co) metal-organic frameworks, and then immobilized on poly(vinylidene fluoride) (PVDF) membrane to construct macroporous Fe-Co@NC-CNTs/PVDF catalytic membrane via directional freezing technique, where active sites were efficiently exposed for oxidants and target pollutants. As expected, Fe-Co@NC-CNTs/PVDF membrane successfully achieved almost 100% bisphenol A (BPA) degradation after 40 min via PMS activation, which was significantly overperformed the majority of conventional carbon-based catalysts. Besides, we found that Fe-Co@NC-CNTs/PVDF membrane not only exhibited ideal catalytic and self-cleaning property in humic acid (HA)-BPA coexistence system, but also maintained the excellent reusability and ultrahigh water flux (10464.45 L m-2 h-1) even after 5 cycles. Notably, in EPR analysis and quenching experiments, it was found that sulfate radicals (SO4·- and ·OH) and singlet oxygen (1O2) participated the degradation process while 1O2 made a major contribution. More significantly, this study is very meaningful for the development of novel catalytic self-cleaning membranes with PMS activation.

12.
J Hazard Mater ; : 124402, 2020 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-33189469

RESUMO

Thallium (Tl) pollution caused by the exploitation of uranium (U) mines has long been neglected due to its low crustal abundance. However, Tl may be enriched in minerals of U ore because Tl has both sulfurophile and lithophile properties. Herein, a semi-dynamic leaching experiment combined with statistical analysis, geochemical speciation and multi-characterization provided novel insight into the distinct features and mechanisms of Tl release from uranium mill tailings (UMT). The results showed that particle size effects prevail over the pH on Tl release, and surface dissolution is the pivotal mechanism controlling Tl release based on Fick's diffusion model. The study revealed that long-term leaching and weathering can lead to the increased acid-extractable and oxidizable fractions of Tl in UMT, and that the exposure and dissolution of Tl-containing sulfides would largely enhance the flux of Tl release. The findings indicate that UMT containing (abundant) pyrite should be paid particular attention due to Tl exposure. Besides, critical concern over the potential Tl pollution in universal U mining and hydrometallurgical areas likewise may need to be seriously reconsidered.

13.
Environ Int ; 146: 106207, 2020 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-33197789

RESUMO

Industrial activities tend to deteriorate adjacent agricultural lands due to accumulation of potentially toxic elements in soils and crops. However, better understanding of their distinctive source partitions and transfer process remains insufficient in steel-making area. The paper focuses on the pollution levels, health risks, and provenance identification of Tl, As, Pb, Cu, Ni, Co, Sb, Cd, Zn, Be, Cr, Fe, Mn, Mo, Sn, and V in common vegetables from different farmlands near a steel-making plant. The results showed that the Tl, As, Pb, Cd, Cr, Cu and Mn were of high-level contamination in soils and generally above the maximum permissible level (MPL). Calculation using hazard quotients (HQ) exhibited that consumption of the studied vegetables may entail significant health risks to residents, especially for children, resulting from the elevated contents of Tl, As and associated toxic elements. Calculation by binary mixing model using Pb isotopic compositions suggested that steel-making activities contributed to 35-80% of the contamination of Pb and As in vegetables. It is necessary to adopt appropriate remediation measures to mitigate the farmland contamination and ensure the food safety of the agricultural products.

14.
Transgenic Res ; 29(5-6): 587-598, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33170439

RESUMO

Precise gene editing of model organisms is required for accurately modeling human diseases and deciphering gene functions. In this study, we used a pair of guide RNAs (sgRNAs), which in vitro transcribed along with other CRISPR RNA components, to generate two cleavage sites flanking pig GJB2 (pGJB2) CDS. By using long single-stranded DNAs (lssDNA) as homology-directed repair (HDR) templates, we efficiently obtained two gene-edited pigs, of which GJB2 CDS replaced with CDSs containing human GJB2 c.235delC mutation and orthologous human p.V37I mutation, respectively. These mutations were commonly observed in patients with hearing loss. Genetic analysis of the two gene-edited pigs showed that the HDR-derived gene-editing efficiency were as high as 80% (4/5) and 50% (2/4), respectively. While no mutation was observed in the group of single cutting with one sgRNA covering the 235th nucleotide C in pGJB2 CDS, using a short single-stranded oligo DNA containing c.235delC mutation as HDR template. Extra experiments proved that the intended mutations were successfully transmitted to offspring or extensively integrated into various tissues including gonad of founder pigs. Our work indicated that the new "double cutting with lssDNA template" gene editing method can expand sgRNA selection scope and avoids direct cutting of gene CDS. Additionally, can introduce precise mutations into mammalian genomic sites, especially those with unavailable proper protospacer sequence or being resistant to gene editing. Moreover, this method can be performed with CRISPR RNA reagents instead of CRISPR ribonucleoproteins applied in previous reports.

15.
J Biol Chem ; 2020 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-33208464

RESUMO

Rabies virus (RABV) matrix protein (M) plays crucial roles in viral transcription, replication, assembly, and budding; however, its function during the early stage of virus replication remains unknown. Here, we mapped the protein interactome between RABV M and human host factors using a proteomic approach, finding a link to the V-type proton ATPase (V-ATPase) catalytic subunit A (ATP6V1A) which is located in the endosomes where RABV first enters. By downregulating or upregulating ATP6V1A expression in HEK293T cells, we found that ATP6V1A facilitated RABV replication. We further found that ATP6V1A was involved in the dissociation of incoming viral M proteins during viral uncoating. Co-immunoprecipitation demonstrated that M interacted with the full length or middle domain of ATP6V1A, which was dependent on the lysine residue at position 256 and the glutamic acid residue at position 279. RABV growth and uncoating in ATP6V1A-depleted cells was restored by trans-complementation with the full length or interaction domain of ATP6V1A. Moreover, stably overexpressed ATP6V1A enhanced RABV growth in Vero cells which are used for the production of rabies vaccine. Our findings identify a new partner for RABV M proteins and establish a new role of ATP6V1A by promoting virion uncoating during RABV replication.

16.
J Food Biochem ; : e13556, 2020 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-33152804

RESUMO

The cellular defense pathway plays a key role in maintaining the homeostasis, tissues and organisms. Nuclear factor E2-related factor 2 (Nrf2), as a key cell signaling pathway, plays an important role in encoding detoxification enzymes and other stress response mediators. Recent studies have shown that it is closely related to the prevention and treatment of acute kidney injury (AKI). Therefore, this article reviews the protective effects of Nrf2-related signaling pathways on acute kidney injury, and summarizes the strategies of natural pharmaceutical ingredients such as flavonoids, alkaloids, terpenes, phenylpropionic acid, polyphenols, and polysaccharides to prevent and treat acute kidney injury. It is of great significance to further study the relationship between Nrf2 regulated signal pathway and kidney disease and the development of new medicines for acute kidney injury treatment. It can also provide new ideas and treatment strategies for clinical treatment of acute kidney injury. PRACTICAL APPLICATIONS: This article reviewed the mechanisms by which the active ingredients of natural medicines slow down acute kidney injury through the Nrf2 pathway. It will help us to understand the regulatory role of the Nrf2 pathway in AKI more comprehensively, and provide a theoretical basis for further exploring the mechanism of more natural drugs to reduce acute kidney injury.

17.
Biomed Pharmacother ; 131: 110796, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33152952

RESUMO

The acoustic radiation forces produced by ultrasonic stimulation induce shear stress on objects in the acoustic field. Piezo1, a mechanosensitive ion channel protein that is expressed on the plasma membranes of vertebrate cells, can sense shear stress and transduce it into downstream signaling. In this study, we examined the sensitivity of Piezo1 to ultrasonic stimulation and assessed its downstream biological functions in human umbilical vein endothelial cells (HUVECs). Ultrasonic stimulation using a stimulation power of 0.2 W and a frequency of 1 MHz for 10 s did not induce cell damage. However, ultrasonic stimulation induced an influx of calcium ions, which increased with an increase in the stimulation duration. Knockdown of Piezo1 protein decreased the influx of calcium ions during ultrasonic stimulation, which demonstrated that Piezo1 may be activated by the shear stress produced by ultrasonic stimulation. The influx of calcium ions in response to ultrasonic stimulation could be modulated by the Piezo1 protein level. Additionally, ultrasonic stimulation reduced the levels of downstream factors such as MLCK and ATP, which are involved in the Ca2+/CaM/MLCK pathway, by suppressing Piezo1. As the Ca2+/CaM/MLCK pathway influences the permeability of the cell membrane, the internalization of FITC-Dextran into cells under ultrasonic stimulation was validated. Ultrasonic stimulation was demonstrated to promote the increase in cell permeability, and the suppression of Piezo1 was shown to induce the decrease in cell permeability. Therefore, this study shows that ultrasonic stimulation may modulate the permeability of the membrane of HUVECs by modulating the expression of Piezo1 protein.

18.
Acta Pharm Sin B ; 10(9): 1769-1783, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33088695

RESUMO

Atherosclerosis (AS) is the leading cause of heart attacks, stroke, and peripheral vascular disease. Berberine (BBR), a botanical medicine, has diversified anti-atherosclerotic effects but with poor absorption. The aim of this study was to develop an effective BBR-entrapped nano-system for treating AS in high-fat diet (HFD)-fed Apoe (-/-) mice, and also explore the possible underlying mechanisms involved. Three d-α-tocopherol polyethylene glycol (PEG) succinate (TPGS) analogues with different PEG chain lengths were synthesized to formulate BBR-entrapped micelles. HFD-fed Apoe (-/-) mice were administered with optimized formula (BBR, 100 mg/kg/day) orally for 5 months. The artery plaque onset and related metabolic disorders were evaluated, and the underlying mechanisms were studied. Our data showed that, BT1500M increased BBR deposition in liver and adipose by 107.6% and 172.3%, respectively. In the Apoe (-/-) mice, BT1500M ameliorated HFD-induced hyperlipidemia and lipid accumulation in liver and adipose. BT1500M also suppressed HFD-induced chronic inflammation as evidenced by the reduced liver and adipose levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß); and decreased plasma level of TNF-α, IL-6, IL-1ß, interferon-γ (IFN-γ), monocyte chemotactic protein (MCP), and macrophage inflammatory factor (MIP). The mechanism study showed that BT1500M changed Ampk and Nf-κb gene expression, and interrupted a crosstalk process between adipocytes and macrophages. Further investigation proved that BT1500M decreased endothelial lesion and subsequent macrophage activation, cytokines release, as well as cholesteryl ester gathering in the aortic arch, resulting in ameliorated artery plaque build-up. Our results provide a practical strategy for treating AS using a BBR-entrapped nano-system.

19.
J Assist Reprod Genet ; 37(12): 3057-3067, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33089439

RESUMO

PURPOSE: This study aimed to determine the effects of drilling and thinning treatment of laser-assisted hatching on the expression and methylation of imprinted gene IGF2/H19 in embryos and offspring. METHODS: The prehatching blastocysts with treatment of drilling or thinning, or control prehatching blastocysts, were transplanted in surrogate uteri. The DNA methylation of IGF2/H19 imprinting control region (ICR) and the expression of IGF2 and H19 were respectively evaluated using bisulfite conversion-mediated sequencing and real-time PCR. RESULTS: The drilling group showed a significant increase in the development rate of hatched blastocysts in comparison with the control and thinning group. DNA methylation level of IGF2/H19 ICR of hatched blastocysts in the thinning group was 27.33% in comparison with the 38.67% and 36% observed in the control and drilling group. The thinning treatment increased the DNA methylation level of IGF2/H19 ICR in the placenta in comparison with the control and drilling group. The drilling and thinning treatment decreased the expression level of H19 mRNA in prehatching and hatched blastocysts as well as placenta, while a significant increase in the expression level of H19 mRNA of offspring was observed in the thinning group. The thinning treatment increased the expression level of IGF2 mRNA of prehatching blastocysts and offspring and a significant decrease in placenta, while the drilling treatment resulted in a significant increase in the expression level of IGF2 mRNA of hatched blastocysts and placenta. CONCLUSION: These observations suggested that drilling used for hatching of in vitro cultured mouse blastocysts may improve the production of offspring.

20.
J Clin Invest ; 130(11): 5951-5966, 2020 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-33016929

RESUMO

ARID1A, a component of the chromatin-remodeling complex SWI/SNF, is one of the most frequently mutated genes in human cancer. We sought to develop rational combination therapy to potentiate the efficacy of immune checkpoint blockade in ARID1A-deficient tumors. In a proteomic analysis of a data set from The Cancer Genomic Atlas, we found enhanced expression of Chk2, a DNA damage checkpoint kinase, in ARID1A-mutated/deficient tumors. Surprisingly, we found that ARID1A targets the nonchromatin substrate Chk2 for ubiquitination. Loss of ARID1A increased the Chk2 level through modulating autoubiquitination of the E3-ligase RNF8 and thereby reducing RNF8-mediated Chk2 degradation. Inhibition of the ATM/Chk2 DNA damage checkpoint axis led to replication stress and accumulation of cytosolic DNA, which subsequently activated the DNA sensor STING-mediated innate immune response in ARID1A-deficient tumors. As expected, tumors with mutation or low expression of both ARID1A and ATM/Chk2 exhibited increased tumor-infiltrating lymphocytes and were associated with longer patient survival. Notably, an ATM inhibitor selectively potentiated the efficacy of immune checkpoint blockade in ARID1A-depleted tumors but not in WT tumors. Together, these results suggest that ARID1A's targeting of the nonchromatin substrate Chk2 for ubiquitination makes it possible to selectively modulate cancer cell-intrinsic innate immunity to enhance the antitumor activity of immune checkpoint blockade.

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