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Bone regeneration is a well-orchestrated process synergistically involving inflammation, angiogenesis, and osteogenesis. Therefore, an effective bone graft should be designed to target multiple molecular events and biological demands during the bone healing process. In this study, a biodegradable gelatin methacryloyl (GelMA)-based Janus microsphere delivery system containing calcium phosphate oligomer (CPO) and bone morphogenetic protein-2 (BMP-2) is developed based on natural biological events. The exceptional adjustability of GelMA facilitates the controlled release and on-demand application of biomolecules, and optimized delivery profiles of CPO and BMP-2 are explored. The sustained release of CPO during the initial healing stages contributes to early immunomodulation and promotes mineralization in the late stage. Meanwhile, the administration of BMP-2 at a relatively high concentration within the therapeutic range enhances the osteoinductive property. This delivery system, with fine-tuned release patterns, induces M2 macrophage polarization and creates a conducive immuno-microenvironment, which in turn facilitates effective bone regeneration in vivo. Collectively, this study proposes a bottom-up concept, aiming to develop a user-friendly and easily controlled delivery system targeting individual biological events, which may offer a new perspective on developing function-optimized biomaterials for clinical use.
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Phospholipids (PL) have garnered significant attention due to their physiological activities. Milk and other dairy products are important dietary sources for humans and have been extensively used to analyze the presence of PL by various analytical techniques. In this paper, the analysis techniques of PL were reviewed with the eight trigrams of phospholipidomics and a comprehensive fingerprint of 1295 PLs covering 8 subclasses in milk and other dairy products, especially. Technology is the primary productive force. Based on phospholipidomics technology, we further review the relationship between the composition of PL and factors that may be involved in processing and experimental operation, and emphasized the significance of the biological role played by PL in dietary supplements and biomarkers (production, processing and clinical research), and providing the future research directions.
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In pharmaceutics, ingredients are classified as active ingredients and excipients. In topical/transdermal phytomedicines, an ingredient may serve both functions. Published information on these dual-purpose ingredients and their pharmacological relevance is limited. An intriguing scenario arises in traditional Chinese medicine (TCM) formulations, where active ingredients and excipients are undifferentiated. This study analyzes ingredients in TCM topical/transdermal formulations, aiming at harmonization of understanding of TCMs. The most commonly recorded ingredients from such formulations in the Chinese pharmacopoeia 2020 (ChP 2020) are reviewed, aiming at developing innovative topical/transdermal phytomedicines. Current editions of Chinese historical documents were reviewed to explore the principles underlying the use of these ingredients. TCM formulations containing botanical drugs for topical/transdermal application were selected from the ChP 2020. The use of botanical materials in TCM formulations is guided by the "Jun-Chen-Zuo-Shi" principle rooted in Yin-Yang and the five elements' theories. In the ChP 2020, 155 botanical drugs, along with 40 excipients (from the "procedure" section, focusing on processing and technical parameters), were identified from 34 botanical formulations intended for topical/transdermal application. Pungent and aromatic botanical materials were the most frequently recorded. Adhesive plasters were the most commonly recorded TCM dosage form, employing specific matrix blends. This new perspective of study reveals the prevalence of pungent and aromatic botanical materials, the common use of adhesive plasters, multifunctional properties of botanical oils, and formulation adaptability in TCM topical/transdermal products. These insights should inform novel formulation designs for both pharmaceutical and phytopharmacological research.
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Central nervous system (CNS)-related diseases have a high mortality rate, are a serious threat to physical and mental health, and have always been an important area of research. Gastrodin, the main active metabolite of Gastrodia elata Blume, used in Chinese medicine and food, has a wide range of pharmacological effects, mostly related to CNS disorders. This review aims to systematically summarize and discuss the effects and underlying mechanisms of gastrodin in the treatment of CNS diseases, and to assess its potential for further development as a lead drug in both biomedicine and traditional Chinese medicine. Studies on the pharmacological effects of gastrodin on the CNS indicate that it may exert anti-neurodegenerative, cerebrovascular protective, and ameliorative effects on diabetic encephalopathy, perioperative neurocognitive dysfunction, epilepsy, Tourette's syndrome, depression and anxiety, and sleep disorders through various mechanisms. To date, 110 gastrodin products have been approved for clinical use, but further multicenter clinical case-control studies are relatively scarce. Preclinical studies have confirmed that gastrodin can be used to treat CNS-related disorders. However, important concerns need to be addressed in the context of likely non-specific, assay interfering effects when gastrodin is studied using in vitro and in silico approaches, calling for a systematic assessment of the evidence to date. High-quality clinical trials should have priority to evaluate the therapeutic safety and clinical efficacy of gastrodin. Further experimental research using appropriate in vivo models is also needed, focusing on neurodegenerative diseases, cerebral ischemic and hypoxic diseases, brain damage caused by methamphetamine or heavy metals, and epilepsy.
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Candida albicans biofilms are responsible for oral candidiasis. Fluoxetine is a widely used antidepressant, with certain anti-Candida activities. The antifungal activity of fluoxetine combined with various antifungals against C. albicans biofilms and oral candidiasis was evaluated in this study. The morphological change in the inhibition of fluoxetine on C. albicans biofilms was observed using SEM. The interactions between fluoxetine and antifungals against C. albicans biofilms were evaluated using microdilution checkerboard methods, FICI and the ΔE model. The synergistic combination was tested in vivo on the mice model of oral candidiasis. SEM imaging showed fluoxetine inhibited hyphal growth and biofilm formation. Fluoxetine combined with caspofungin exhibited synergistic effects against C. albicans biofilms. Antagonistic effects occurred when fluoxetine was combined with amphotericin B or terbinafine. Further, the fluoxetine combined with caspofungin significantly reduced the lesion score and CFU of C. albicans on the murine tongue (p < 0.05), and relieved oral candidiasis of the infected mice.
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Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans , Candidíase Bucal , Fluoxetina/farmacologia , Animais , Camundongos , Testes de Sensibilidade MicrobianaRESUMO
Based upon the intramolecular charge transfer (ICT) mechanism, a novel ratiometric fluorescent probe EB was developed to detect SO32-/HSO3-. The probe displayed both colorimetric and ratiometric responses toward SO32-/HSO3-. It displayed a quick response (within 60 s), good selectivity and high sensitivity (a detection limit of 28 nM) towards SO32-/HSO3-. The SO32-/HSO3- sensing mechanism was confirmed as the Michael addition reaction by ESI-MS. Moreover, the probe could be applied to measure the level of sulfite in real samples, like sugar and chrysanthemum, and it could also be used to detect SO32-/HSO3- in HepG2 cells through confocal fluorescence microscopy, which proved its practical application in clinical diagnosis.
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Carbocianinas/química , Corantes Fluorescentes/química , Dióxido de Enxofre/química , Linhagem Celular Tumoral , Colorimetria/métodos , Fluorescência , Células Hep G2 , Humanos , Limite de Detecção , Sensibilidade e Especificidade , Sulfitos/químicaRESUMO
We investigated the effect of reduced contrast on speed perception for two types of tasks: (a) the speed of a rotating image, an example of "object-motion," and (b) speed of travel when viewing wide-screen videos recorded from inside a car, an example of "self-motion." Both types of stimuli were presented over a range of spatial contrasts. The results showed that reduced contrast caused significant decreases of perceived speed for the rotating disk, replicating the well known Thompson Effect. Reduced contrast had inconsistent effects on perceived speed of self-motion, however, resulting in perception of faster self-motion at the lowest speed, slower self-motion at higher speeds, and no effect at intermediate speed. Although further research is needed, the differential effects of reduced contrast on perceived speed of object-motion vs. self-motion are consistent with evidence for two modes of vision.
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Percepção de Movimento/fisiologia , Movimento , Percepção Visual/fisiologia , Condução de Veículo , Sensibilidades de Contraste , Humanos , Ilusões Ópticas/fisiologia , Gravação em VídeoRESUMO
As the most prevalent mycotoxin in agricultural products, aflatoxin B1 not only causes significant economic losses but also poses a substantial threat to human and animal health. AFB1 has been shown to increase the risk of hepatocellular carcinoma (HCC) but the underlying mechanism is not thoroughly researched. Here, we explored the toxicity mechanism of AFB1 on human hepatocytes following low-dose exposure based on transcriptomics and lipidomics. Apoptosis-related pathways were significantly upregulated after AFB1 exposure in all three hES-Hep, HepaRG, and HepG2 hepatogenic cell lines. By conducting a comparative analysis with the TCGA-LIHC database, four biomarkers (MTCH1, PPM1D, TP53I3, and UBC) shared by AFB1 and HCC were identified (hazard ratio > 1), which can be used to monitor the degree of AFB1-induced hepatotoxicity. Simultaneously, AFB1 induced abnormal metabolism of glycerolipids, sphingolipids, and glycerophospholipids in HepG2 cells (FDR < 0.05, impact > 0.1). Furthermore, combined analysis revealed strong regulatory effects between PIK3R1 and sphingolipids (correlation coefficient > 0.9), suggesting potential mediation by the phosphatidylinositol 3 kinase (PI3K) /protein kinase B (AKT) signaling pathway within mitochondria. This study revealed the dysregulation of lipid metabolism induced by AFB1 and found novel target genes associated with AFB-induced HCC development, providing reliable evidence for elucidating the hepatotoxicity of AFB as well as assessing food safety risks.
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Goat milk is considered the optimal substitute for human milk and is characterized by variations in the lipid composition of its fat globules across lactation phases. Therefore, the objective of this study was to thoroughly analyze the differences between goat milk during different lactations and human milk, aiming to offer scientific guidance for the production of functional dairy products. Compared with transitional and mature milk, the findings indicated that the total membrane protein content in goat colostrum exhibited greater similarity to that found in human milk. Additionally, goat milk exhibited higher milk fat globule size, as well as a higher total lipid and protein content than human milk. A total of 1461 lipid molecules across 61 subclasses were identified in goat milk and human milk. The contents of glycerides and glycerophospholipids were higher in goat colostrum, whereas sphingolipids and fatty acids were more abundant in human milk. Meanwhile, the compositions of lipid subclasses were inconsistent. There were 584 differentially expressed lipids identified between human and goat milk, including 47 subclasses that were primarily involved in the metabolism of glycerophospholipids, sphingolipids, and triglycerides. In summary, for both the membrane protein and the lipid composition, there were differences between the milk of different goat lactations and human milk.
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Fate and transport of nanoplastics in aquatic environments are affected by their heteroaggregation with minerals in the presence of macromolecules. This study investigated the heteroaggregation of polystyrene nanoplastics (PSNPs) with goethite nanoparticles (GNPs) under the influence of macromolecules [humic acid (HA), bovine serum albumin (BSA), and DNA] and electrolytes. Under 1 mg C/L macromolecule, raising electrolyte concentration promoted heteroaggregation via charge screening, except that calcium bridging with HA also enhanced heteroaggregation at CaCl2 concentration above 5 mM. At all NaCl concentrations and CaCl2 concentration below 5 mM, 1 mg C/L macromolecules strongly retarded heteroaggregation, ranking BSA > DNA > HA. Raising macromolecule concentration strengthened such stabilization effect of all macromolecules in NaCl solution and that of DNA and BSA in CaCl2 solution by enhancing steric hindrance. However, 0.1 mg C/L BSA slightly promoted heteroaggregation in CaCl2 solution due to stronger electrostatic attraction than steric hindrance. In CaCl2 solution, raising HA concentration strengthened its destabilization effect via calcium bridging. Macromolecules having more compact globular structure and higher molecular weight may exert greater steric hindrance to inhibit heteroaggregation more effectively. This study provides new insights on the effects of macromolecules and electrolytes on heteroaggregation between nanoplastics and iron minerals in aquatic environments.
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Eletrólitos , Compostos de Ferro , Minerais , Nanopartículas , Poliestirenos , Poluentes Químicos da Água , Poliestirenos/química , Minerais/química , Eletrólitos/química , Compostos de Ferro/química , Nanopartículas/química , Poluentes Químicos da Água/química , Cinética , DNA/química , DNA/efeitos dos fármacos , Soroalbumina Bovina/química , Cloreto de Cálcio/químicaRESUMO
PURPOSE: Apically extruded debris, canal transportation and shaping ability were compared between contracted endodontic cavities (CECs) and traditional endodontic cavities (TECs) after instrumentation with XP-endo Shaper (XPS), ProTaper Gold (PTG), ProTaper for hand-use (HPT) and Hero Shaper. METHODS: The CECs or TECs groups were sub-divided into 24 groups according to root canal morphology and nickel-titanium (Ni-Ti) instruments. The weight of apically extruded debris was calculated using the Myers and Montgomery model. Pre- and postoperative images of teeth were scanned using micro-CT and the three-dimensional models were constructed and compared. RESULTS: Under CECs or TECs, XPS and PTG produced less apical debris and formed less canal transportation than HPT and Hero Shaper (P < 0.05). XPS group under CECs extruded less apical debris than that under TCEs for round canals with curvature of 20°-35° (P < 0.05). The centering ratios of four tested instruments were higher under TECs than those under CECs (P < 0.05). The HPT and Hero Shaper had more transportation under CECs than that under TCEs (P < 0.05). No statistical difference was found regarding shaping ability among all the groups. CONCLUSION: Under CECs, XPS preserves the original root canal anatomy, meanwhile it produces less apical debris than the other instruments.
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Níquel , Titânio , Cavidade Pulpar/cirurgia , Cavidade Pulpar/anatomia & histologia , Preparo de Canal Radicular , Dente Molar/cirurgia , Desenho de EquipamentoRESUMO
Mycotoxin aflatoxin B1 (AFB1) has been proven to cause neurotoxicity, but its potential interference with the normal function of brain tissue is not fully defined. As the indispensable role of lipids in maintaining the normal function of brain tissue, the aim of this study is to clarify the effect of AFB1 short-term (7 days) exposure on brain tissue from the perspective of lipid metabolism. In this study, zebrafish were exposed to two concentrations (5, 20 µg/L). Through quantitative analysis of AFB1, the detection of AFB1 in zebrafish brain tissue was discovered for the first time, combined with the changes in zebrafish neurobehavior, the occurrence of brain injury was deduced. Subsequently, 1734 lipids in zebrafish brain tissue were mapped using ion mobility time-of-flight mass spectrometry (UPLC-QTOF-IMS-MS), which has great advantages in lipid detection. Comparative analysis of the abnormal lipid metabolism in zebrafish brain revealed 114 significantly changed lipids, mainly involving two pathways of sphingolipid metabolism and fatty acid degradation. This study discovered the detection of AFB1 in the brain and revealed a potential link between AFB1-induced behavioral abnormalities and lipid metabolism disorders in brain tissue, providing reliable evidence for elucidating the neurotoxicity of AFB1.
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Micotoxinas , Peixe-Zebra , Animais , Peixe-Zebra/metabolismo , Aflatoxina B1/toxicidade , Lipidômica , Micotoxinas/metabolismo , LipídeosRESUMO
The effects of antibiotic contamination on vegetable safety and the ecological risks of soil after returning livestock and poultry manure to the land require sufficient future attention. Tetracycline antibiotics (TCs) are often detected at high concentrations in livestock manure and vegetable production soils. Recently, pot experiments and field investigation methods have often been used to understand the effects of TCs contamination on the vegetable safety and ecological risks of soil, whereas field experiments are employed less frequently. This study investigated the distribution characteristics of TCs in the soil-vegetable system following manure application using a combination of pot and field experiments. The human health risks of the edible parts of Chinese flowing cabbage were assessed using the health risk quotient method based on the acceptable daily intake (ADI-HQ), and the ecological risks of TCs-contaminated soils were evaluated using the risk quotient method associated with the species sensitivity distribution model (SSD-RQ). The results showed that oxytetracycline (OTC) was the major type of TCs in Chinese flowering cabbage based on both the pot and field experiments. The maximum contents (dry weight) of OTC in the aboveground parts of the Chinese flowering cabbage for the pot and field experiments were 29.25 µg·kg-1 and 45.03 µg·kg-1, respectively, whereas those of their underground parts were 87.32 µg·kg-1 and 135.44 µg·kg-1, respectively. Meanwhile, higher contents of TCs were detected in Chinese flowering cabbage collected from the field experiment than those from the pot experiment. OTC was also the major type of TCs in soil from both the pot and field experiments, with their contents up to 604.30 µg·kg-1 and 1013.68 µg·kg-1, respectively. Higher residual contents of three TCs were detected in soils collected from the field experiment than those from the pot experiment. Under the experimental conditions, with the except that OTC in Chinese flowering cabbage from the field experiment would pose medium health risks (HQ>0.1) to children, the contents of three TCs in other treated Chinese flowering cabbage would pose low health risks (HQ ≤ 0.1) to adults and children. In the pot experiments, three TCs present in Chinese flowering cabbage would pose low health risks (HQ ≤ 0.1) to adults and children. Additionally, the TCs in soils with manure application from the pot and field experiments may have posed both moderate or high levels of ecological risks (HQ>0.1 or HQ>1). Therefore, the effects of antibiotic contamination on vegetable safety and their potential ecological risks on soil following manure fertilization need to be given special attention.
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Oxitetraciclina , Verduras , Adulto , Animais , Criança , Humanos , Esterco , Antibacterianos , Medição de Risco , Gado , Solo , FertilizaçãoRESUMO
Epidemiological studies have suggested an association between obesity and periodontal disease. Brown adipose tissue (BAT) has an anti-obesity effect. However, the effects of periodontitis on obesity and BAT remain unclear. Therefore, the present study aimed to determine the effects of lipopolysaccharide derived from Porphyromonas gingivalis (P. gingivalis LPS) on brown adipocytes. For this purpose, the present study examined the effects of the intravenous administration of Porphyromonas gingivalis (P. gingivalis) in mice, the treatment of brown adipocytes with P. gingivalis LPS during differentiation, and the administration of small interfering RNA targeting interferon on brown preadipocytes by assessing the expression of genes involved in differentiation, using a long non-coding (lnc)RNA, and pro-inflammatory factors using reverse transcription-quantitative PCR. In addition, the accumulation of lipid droplets was examined using Oil Red O staining. P. gingivalis LPS reduced the expression of uncoupling protein 1 (UCP1) and lncRNA-BATE10 in brown adipocytes during differentiation. Consistent with this finding, P. gingivalis reduced UCP1 and lncRNA-BATE10 expression in the BAT of mice. lncRNA-BATE10 may thus be involved in the regulation of UCP1 expression that occurs during the differentiation of brown adipocytes treated with P. gingivalis LPS. Thus, P. gingivalis LPS may inhibit BAT differentiation by reducing lncRNA-BATE10 expression.
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Tetracycline antibiotics (TCs) introduced into agricultural fields via manure application tend to accumulate in soils and further reach water environments via surface runoff and leachate, posing potential risks to regional water environment. This study investigated the loss of tetracycline (TC), oxytetracycline (OTC), and chlortetracycline (CTC) in surface runoff and leachate samples collected from a vegetable farmland with manure application in Guangzhou, South China. A risk assessment method was constructed for evaluating the ecological and health risks of manure-associated antibiotics released from soil into water environment. The results showed that the concentrations of three TCs in surface runoff, 30-cm leachate, and 60-cm leachate after the first rainfall event were 2.79-35.97, 1.71-18.44, and 0.4-2.66 µg/L, respectively, which all decreased with sampling depth and the time after rainfall events. Up to 0.13% of TCs were transported into the surface water through surface runoff, while less than 0.01% of TCs were transported into the groundwater through leachate at 60 cm. OTC had a higher total mass percentage (0.13%) into surface water via runoff than CTC (0.11%) and TC (0.07%) likely due to its smallest Kd value and largest input mass. Based on loss percentages, their predicted environmental concentrations (PEC) ranged from 4.87 (TC) to 16.91 (OTC) ng/L in regional surface water and 1.42 (TC) to 5.20 (CTC) ng/L in regional groundwater. The risk assessment based on PEC results suggested non-negligible health risk (HQ > 1.0 × 10-6) and low ecological risk (RQ < 0.1) in both regional surface water and groundwater, drawing concerns on the potential hazards of TCs released from manure-amended soil into water environments.
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Clortetraciclina , Oxitetraciclina , Poluentes do Solo , Antibacterianos/análise , Clortetraciclina/análise , Esterco , Medição de Risco , Solo , Poluentes do Solo/análise , Tetraciclina , ÁguaRESUMO
The present study was designed to explore the in vivo-antioxidant capacity and the probable mechanism of AAPs-H, prepared from Auricularia auricula polysaccharides with the optimal extraction conditions by Box-Behnken design and acid hydrolysis, using Caenorhabditis elegans as a model organism. The effects of AAPs-H on the locomotion behavior, life span, antioxidant-related enzymes activities, and antioxidants levels in C. elegans were studied. Furthermore, the potentials of AAPs-H in up-regulating the expression of antioxidant-related genes in C. elegans, such as skn-1, sod-3 and sir-2.1, were also discussed. AAPs-H demonstrated a highly significant protective effect against the damage caused by paraquat, could significantly increase U-Turn frequency of worms (p < 0.01), extend their lifespan, enhance antioxidant systems including GR by 63.96% (p < 0.05), GSH-Px by 71.16% (p < 0.01), SOD by 78.65% (p < 0.01) and CAT by 98.52% (p < 0.01), increase the level of GSH by 28.12% (p < 0.05), and decrease the level of MDA by 39.29% (p < 0.01). The qRT-PCR results showed that AAPs-H could up regulate mRNA expression levels of skn-1, sod-1, sod-2, sod-3 and sir-2.1 in wild-type C. elegans (>1.6 fold) when treated with the concentration of 0.4 mg/mL (p < 0.05 or p < 0.01). Our studies provide evidence that AAPs-H improves antioxidant defense system, and up-regulation of oxidative stress related genes for prevention of stress damage in C. elegans.
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Auricularia/química , Produtos Biológicos/isolamento & purificação , Produtos Biológicos/farmacologia , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/fisiologia , Polissacarídeos Fúngicos/isolamento & purificação , Polissacarídeos Fúngicos/farmacologia , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Produtos Biológicos/química , Fracionamento Químico/métodos , Polissacarídeos Fúngicos/química , Regulação da Expressão Gênica/efeitos dos fármacos , Hidrólise , Locomoção/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Paraquat/química , Paraquat/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Temperatura , Ondas UltrassônicasRESUMO
Arsenic trioxide (ATO), an ancient traditional Chinese medicine, has been successfully used as a therapeutic agent for leukemia. Drug resistance and toxicity are major concerns with the treatment. MicroRNAs (miRNAs) are endogenous small non-coding RNA molecules that might modulate cellular sensitivity to anticancer drugs. miRNA-21 (miR-21) is one of the most prominent miRNAs involved in various aspects of human cancers. However, miR-21 has been rarely characterized in chronic myelogenous leukemia (CML). Here, we used a specific anti-miR-21 oligonucleotide (AMO-miR-21) to sensitize K562 cells to ATO by degradation of miR-21. The results showed that both AMO-miR-21 and ATO caused growth inhibition, apoptosis, and G1-phase arrest in K562 cells. Meanwhile, AMO-miR-21 significantly promoted ATO-mediated growth inhibition and apotosis without affecting the G1 phase. Apoptotic cells were confirmed morphologically with Giemsa's staining. Furthermore, dual-luciferase reporter vector, containing two tandem miR-21 binding sites from PDCD4 3'UTR, validated that PDCD4 was directly regulated by miR-21. Therefore, AMO-miR-21 sensitized leukemic K562 cells to ATO by inducing apoptosis partially due to its up-regulation of PDCD4 protein level. The combination of ATO and AMO-miR-21 present therapeutic potential for CML.
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Antineoplásicos/farmacologia , Arsenicais/uso terapêutico , Leucemia Mielogênica Crônica BCR-ABL Positiva/terapia , MicroRNAs/farmacologia , Oligonucleotídeos/farmacologia , Óxidos/uso terapêutico , Apoptose , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Trióxido de Arsênio , Ciclo Celular/genética , Fase G1/genética , Humanos , Células K562 , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Regulação para CimaRESUMO
OBJECTIVE: To study the effect of antisense oligonucleotide targeted on miRNA-21 (AMO-miR-21) for enhancing the arsenic trioxide (As2O3) sensitivity of leukemic K562 cells and its possible acting mechanism. METHODS: Chemosynthetic AMO-miR-21 was transfected to K562 cells using Lipofectamine TM 2000. The inhibitory effects of As2O3 and AMO-miR-21, used singly or in combining, on cell proliferation were detected by MTT, their inhibition rate and IC50 were calculated. Cell cycle and apoptosis were assessed with PI stain; expression of miRNA-21 in cells was detected quantitatively by real-time PCR, and the potential target gene PDCD, protein expression was detected by immuno-fluorimetry. RESULTS: Used in combining with AMO-miR-21, the IC50 of As2O3, could be lowered from 2.1 micromol/L to 1.23 micromol/L, and the sensitivity of cells to As2O3 increased to 1.78-fold; with the amount of apoptotic cells increased significantly. Transfection with AMO-miR-21 alone could downregulate the expression of miRNA-21 in cells (P < 0.01), and up-regulate PDCD, protein expression level significantly. CONCLUSIONS: Combined use of AMO-miR-21 and As2O3 could increase the sensitivity of K562 cells to As2O3, which provides a novel potential approach for treatment of leukemia. AMO-miR-21 realizes it anti-tumor action by way of targeted inhibition on miRNA-21, and further up-regulates the expression of anti-tumor gene PDCD4.
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Antineoplásicos/farmacologia , Arsenicais/farmacologia , MicroRNAs/efeitos dos fármacos , Óxidos/farmacologia , Proteínas Reguladoras de Apoptose/metabolismo , Trióxido de Arsênio , Regulação Leucêmica da Expressão Gênica , Humanos , Células K562 , Oligonucleotídeos Antissenso/farmacologia , Proteínas de Ligação a RNA/metabolismo , TransfecçãoRESUMO
To explore how hypochlorous acid (HClO) affects human health, a highly sensitive, selective, and trace detection method for hypochlorite (ClO-) is crucial for determining its non-negligible function in both environment and living systems. Herein, a dicyanoisophorone-phenylboronic acid-based novel ratiometric near-infrared fluorescent probe (Probe 1) was designed for the rapid and specific detection of ClO- based on the intramolecular charge transfer (ICT) mechanism. Excess addition of HClO to the Probe 1 solution, 186-times ratio (I652/I582) augment were gained. And this probe provided a colorimetric and ratiometric fluorescence response to ClO- with a high selectivity, a rapid response (within 30â¯s), and had an extremely low detection limit (15.7â¯nM). In addition, owing to the good sensing properties and low cytotoxicity of Probe 1, it can be used to expediently visualize exogenous ClO- in HepG2 cells and endogenous ClO- in RAW264.7 macrophage cells. Furthermore, the probe was successfully used for the bioimaging of zebrafish with an acute inflammation. Thus, Probe 1 is a promising vehicle to identify the level of HClO in animals with associated diseases.
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Ácidos Borônicos/química , Corantes Fluorescentes/química , Ácido Hipocloroso/análise , Inflamação/metabolismo , Nitrilas/química , Animais , Ácidos Borônicos/síntese química , Ácidos Borônicos/toxicidade , Colorimetria/métodos , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/toxicidade , Células Hep G2 , Humanos , Ácido Hipocloroso/metabolismo , Inflamação/induzido quimicamente , Limite de Detecção , Lipopolissacarídeos , Camundongos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Nitrilas/síntese química , Nitrilas/toxicidade , Células RAW 264.7 , Peixe-ZebraRESUMO
We employed a single factor and response surface methodology based on Box-Behnken design (BBD) to optimize the extraction of pumpkin polysaccharides. We then simulated pumpkin polysaccharide gastrointestinal digestion in vitro and investigated their biodistribution in mice. The optimal extraction conditions, with a yield of 7.38⯱â¯0.21%, were as follows: a concentration of NaOH 1.20%, a ratio of material to liquid of 1:11, and an extraction time of 2.1â¯h, respectively, according to single factor and BBD experiments. In the gastrointestinal experiment in vitro, the molecular weight of the polysaccharides markedly decreased after gastric digestion for 30â¯min, suggesting the decline is due to the breakdown of polysaccharide glycosidic bonds. The simulated intestinal fluid had little effect on polysaccharides digestion within 240â¯min. Analysis of the biodistribution in mice indicated that the polysaccharides distribute in the duodenum, jejunum, and ileum 30 to 60â¯min after intragastrical administration, and are absorbed in the jejunum and ileum after 60 to 360â¯min. These results provide information on the digestion and biodistribution of pumpkin polysaccharides and offer a theoretical basis for further understanding the absorption mechanisms in vivo.