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Aqueous zinc ion batteries (AZIBs) have emerged as a promising battery technology due to their excellent safety, high capacity, low cost, and eco-friendliness. However, the cycle life of AZIBs is limited by severe side reactions and zinc dendrite growth on the zinc electrode surface, hindering large-scale application. Here, an electrolyte optimization strategy utilizing the simplest dipeptide glycylglycine (Gly-Gly) additive is first proposed. Theoretical calculations and spectral analysis revealed that, due to the strong interaction between the amino group and Zn atoms, Gly-Gly preferentially adsorbs on zinc's surface, constructing a stable and adaptive interfacial layer that inhibits zinc side reactions and dendrite growth. Furthermore, Gly-Gly can regulate zinc ion solvation, leading to a deposition mode shift from dendritic to lamellar and limiting two-dimensional dendrite diffusion. The symmetric cell with the addition of a 20 g/L Gly-Gly additive exhibits a cycle life of up to 1100 h. Under a high current density of 10 mA cm-2, a cycle life of 750 cycles further demonstrates the reliable adaptability of the interfacial layer. This work highlights the potential of Gly-Gly as a promising solution for improving the performance of AZIBs.
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PURPOSE: To delineate the 10 years' trend regarding Taiwanese adolescents' health perspectives and compare the differences of six adolescent health aspects between Taiwan and the U.S. METHODS: The anonymous structured questionnaire was done every other year with representative sampling methods as Youth Risk Behavior Surveillance System in the United States. Twenty-one questions from six health aspects were extracted for further analysis. Multivariate regression analysis was performed to delineate the relationship among protective factors and risk-taking behaviors, respectively. RESULTS: Overall, 22,419 adolescents were recruited. There were decreasing trend in terms of risk-taking behaviors, such as early contact to pornography (< age 16) (70.6%-60.9%), early cigarette use (< age 13) (20.7%-14.0%), and seriously considering suicide (36.0%-17.8%). There was an increasing trend in behaviors harmful to health: current alcohol drinkers (18.9%-23.4%), and staying up late every day (15.2%-18.5%). Multivariate regression analysis after adjusting gender and grade; it disclosed an increasing trend in protective assets, such as having multiple intimate friends (75.8%-79.3%), satisfaction to body weight and body shape (31.5%-36.1% and 34.5%-40.7%), as well as always wearing a helmet while riding a bike (1.8%-3.0%). CONCLUSION: We should continuously monitor the health status trend of the adolescents to provide them with a healthier environment and well-being.
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Comportamento do Adolescente , Humanos , Adolescente , Estados Unidos , Comportamentos Relacionados com a Saúde , Comportamento Sexual , Assunção de Riscos , EstudantesRESUMO
In recent years, improving plants' resistance towards abiotic stresses with exogenous application of plant growth regulators and nutrients has emerged as a matter of great interest. The present study assessed the potential roles of kinetin (Kn, 0.2 mM) and calcium (Ca, 2 mM) in mitigating the salt (200 mM NaCl) induced inhibitory effects on seed germination and growth of choysum seedlings. The results indicated that NaCl stress significantly reduced the seed germination percentage (42.6%), germination potential (42.0%), germination index (52.1%), seedling vigor index (65.2%), and declined the fresh weight (43.8%), dry weight (52.2%), radicle length (37.2%), and plumule length (41.2%) of germinated seeds, compared to control treatment. The delayed germination and decrease in seedling growth were positively correlated with salinity-induced hormonal imbalance, ion toxicity, and oxidative stress. However, Kn and Ca pretreatment partially mitigated the adverse effects of NaCl stress, evident by early germination and enhanced seedling growth. Kn and Ca effectively increased the accumulation of proline, soluble protein, and soluble sugars, and upregulated the activities of superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase that significantly reduced the production of malondialdehyde, hydrogen peroxide, and superoxide anions in germinating seeds, thereby minimizing the NaCl-induced oxidative damages. Moreover, Kn and Ca pretreatment counteracted the NaCl-induced ionic toxicity by decreasing Na+ and increasing K+ contents and maintained a balanced Na+/K+ ratio in radicles and plumules of choysum seeds. Additionally, Kn and Ca under NaCl stress enhanced hormonal regulation by decreasing the ABA levels with a concomitant increase of GAs (especially GA4) levels and promoted early germination. Remarkably, the co-application of Kn and Ca was most effective by completely counteracting the inhibitory effects of NaCl and maintaining seed germination kinetics, seedling growth, and biochemical parameters almost similar to that in the stress-free control treatment. These results demonstrate that supplementation of Kn and Ca on choysum seeds is an effective chemical strategy regulating the various physiological and biochemical responses that would result in better germination and growth of seeds under stress conditions.
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Brassica rapa , Plântula , Antioxidantes , Cálcio , Germinação , Cinetina/farmacologia , Sementes , Cloreto de SódioRESUMO
The cultivation of leafy vegetables on metal contaminated soil embodies a serious threat to yield and quality. In the present study, the potential role of exogenous jasmonic acid (JA; 0, 5, 10, and 20 µM) on mitigating chromium toxicity (Cr; 0, 150, and 300 µM) was investigated in choysum (Brassica parachinensis L.). With exposure to increasing Cr stress levels, a dose-dependent decline in growth, photosynthesis, and physio-biochemical attributes of choysum plants was observed. An increase in Cr levels also resulted in oxidative stress closely associated with higher lipoxygenase activity (LOX), hydrogen peroxide (H2O2) generation, lipid peroxidation (MDA), and methylglyoxal (MG) levels. Exogenous application of JA alleviated the Cr-induced phytotoxic effects on photosynthetic pigments, gas exchange parameters, and restored growth of choysum plants. While exposed to Cr stress, JA supplementation induced plant defense system via enhanced regulation of antioxidant enzymes, ascorbate and glutathione pool, and the glyoxalase system enzymes. The coordinated regulation of antioxidant and glyoxalase systems expressively suppressed the oxidative and carbonyl stress at both Cr stress levels. More importantly, JA restored the mineral nutrient contents, restricted Cr uptake, and accumulation in roots and shoots of choysum plants when compared to the only Cr-stressed plants. Overall, the application of JA2 treatment (10 µM JA) was more effective and counteracted the detrimental effects of 150 µM Cr stress by restoring the growth and physio-biochemical attributes to the level of control plants, while partially mitigated the detrimental effects of 300 µM Cr stress. Hence, JA application might be considered as an effective approach for minimizing Cr uptake and its detrimental effects in choysum plants grown on contaminated soils.
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Antioxidantes/farmacologia , Brassica/fisiologia , Cromo/toxicidade , Ciclopentanos/farmacologia , Oxilipinas/farmacologia , Poluentes do Solo/toxicidade , Antioxidantes/metabolismo , Ácido Ascórbico/metabolismo , Brassica/efeitos dos fármacos , Brassica/metabolismo , Glutationa/metabolismo , Peróxido de Hidrogênio/metabolismo , Oxirredução , Estresse Oxidativo/fisiologia , Fotossíntese/efeitos dos fármacos , Folhas de Planta/metabolismoRESUMO
BACKGROUND: Interferon alpha (IFNα) is a well-established regulator of immunosuppression in head and neck squamous cell carcinoma (HNSCC), while the role of long noncoding RNAs (lncRNAs) in immunosuppression remains largely unknown. METHODS: Differentially expressed lncRNAs were screened under IFNα stimulation using lncRNA sequencing. The role and mechanism of lncRNA in immunosuppression were investigated in HNSCC in vitro and in vivo. RESULTS: We identified a novel IFNα-induced upregulated lncRNA, lncMX1-215, in HNSCC. LncMX1-215 was primarily located in the cell nucleus. Ectopic expression of lncMX1-215 markedly inhibited expression of the IFNα-induced, immunosuppression-related molecules programmed cell death 1 ligand 1 (PD-L1) and galectin-9, and vice versa. Subsequently, histone deacetylase (HDAC) inhibitors promoted the expression of PD-L1 and galectin-9. Binding sites for H3K27 acetylation were found on PD-L1 and galectin-9 promoters. Mechanistically, we found that lncMX1-215 directly interacted with GCN5, a known H3K27 acetylase, to interrupt its binding to H3K27 acetylation. Clinically, negative correlations between lncMX1-215 and PD-L1 and galectin-9 expression were observed. Finally, overexpression of lncMX1-215 suppressed HNSCC proliferation and metastasis capacity in vitro and in vivo. CONCLUSIONS: Our results suggest that lncMX1-215 negatively regulates immunosuppression by interrupting GCN5/H3K27ac binding in HNSCC, thus providing novel insights into immune checkpoint blockade treatment.
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Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias de Cabeça e Pescoço/patologia , Histonas/metabolismo , Interferon-alfa/farmacologia , RNA Longo não Codificante/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/secundário , Acetilação , Animais , Antivirais/farmacologia , Apoptose , Antígeno B7-H1/genética , Antígeno B7-H1/metabolismo , Biomarcadores Tumorais/genética , Movimento Celular , Proliferação de Células , Galectinas/genética , Galectinas/metabolismo , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Neoplasias de Cabeça e Pescoço/imunologia , Neoplasias de Cabeça e Pescoço/metabolismo , Histonas/química , Histonas/genética , Humanos , Terapia de Imunossupressão , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/tratamento farmacológico , Carcinoma de Células Escamosas de Cabeça e Pescoço/imunologia , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismo , Ativação Transcricional , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto , Fatores de Transcrição de p300-CBP/genética , Fatores de Transcrição de p300-CBP/metabolismoRESUMO
Despite the recognition of the lethality of cancer metastasis and the importance of developing specific anti-metastasis therapies directed at the cancer metastatic cascade, the dynamics of cancer metastasis remains poorly understood. In this study, we examined the dynamics of circulating tumor cell (CTC) survival in the bloodstream using experimental mouse models. CTCs were arrested in the capillaries by adhesion to vascular endothelium within a few minutes after injection into the bloodstream. The loss of CTCs from the circulation followed a bi-phasic decay pattern, with the number of CTCs in the bloodstream being closely associated with the number of blood circulation cycles. The calculated in vivo Vd (apparent volume of distribution) of the CTC revealed organ specific binding of the CTCs. Moreover, confocal microscopy, in vivo fluorescence imaging in syngeneic mouse metastatic models and analysis of blood circulation patterns support the notion of organ-specific tumor metastasis. The present study suggests that organ-specific tumor metastasis is influenced by cooperation between blood circulation patterns and 'seed-soil' compatibility factors. These new findings provide further insights for optimized cancer metastatic prevention strategies such as by creating a hostile circulation microenvironment and targeting the organ-specific 'seed-soil' compatibility factors.
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Carcinoma Pulmonar de Lewis/genética , Melanoma Experimental/genética , Metástase Neoplásica/genética , Células Neoplásicas Circulantes , Animais , Carcinoma Pulmonar de Lewis/patologia , Adesão Celular/genética , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Humanos , Melanoma Experimental/patologia , Camundongos , Metástase Neoplásica/patologia , Especificidade de Órgãos/genética , Microambiente Tumoral/genéticaRESUMO
Salinity represents a serious environmental threat to crop production and by extension, to world food supply, social and economic prosperity of the developing world. Salicylic acid (SA) is an endogenous plant signal molecule involved in regulating various plant responses to stress. In the present study, we characterized the regulatory role of exogenous SA for their ability to ameliorate deleterious effects of salt stress (0, 100, 150, 200â¯mM NaCl) in choysum plants through coordinated induction of antioxidants, ascorbate glutathione (AsA-GSH) cycle, and the glyoxalase enzymes. An increase in salt stress dramatically declined root and shoot growth, leaf chlorophyll and relative water content (RWC), subsequently increased electrolyte leakage (EL) and osmolytes accumulation in choysum plants. Salt stress disrupted the antioxidant and glyoxalase defense systems which persuaded oxidative damages and carbonyl toxicity, indicated by increased H2O2 generation, lipid peroxidation, and methylglyoxal (MG) content. However, application of SA had an additive effect on the growth of salt-affected choysum plants, which enhanced root length, plant biomass, chlorophyll contents, leaf area, and RWC. Moreover, SA application effectively eliminated the oxidative and carbonyl stress by improving AsA and GSH pool, upregulating the activities of antioxidant enzymes and the enzymes associated with AsA-GSH cycle and glyoxalase system. Overall, SA application completely counteracted the salinity-induced deleterious effects of 100 and 150â¯mM NaCl and partially mediated that of 200â¯mM NaCl stress. Therefore, we concluded that SA application induced tolerance to salinity stress in choysum plants due to the synchronized increase in activities of enzymatic and non-enzymatic antioxidants, enhanced efficiency of AsA-GSH cycle and the MG detoxification systems.
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Ácido Ascórbico/genética , Brassica rapa/efeitos dos fármacos , Brassica rapa/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Glutationa/genética , Ácido Salicílico/farmacologia , Estresse Salino/efeitos dos fármacos , Antioxidantes/metabolismo , Ácido Ascórbico/metabolismo , Brassica rapa/crescimento & desenvolvimento , Glutationa/metabolismo , Peróxido de Hidrogênio/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Aldeído Pirúvico/metabolismoRESUMO
In this study, the phytochemical profiles, total and cellular antioxidant activities of five different Chinese chestnut (Castanea mollissima BL.) cultivars were analyzed. Phenolics, flavonoids as well as phytochemical compounds in five cultivars of chestnut kernels were determined. Results showed that the free forms played a dominant role in total phenolics, flavonoids and antioxidant activities of all five cultivars of chestnut kernels. The cultivar 'Fyou' showed the highest total and free phenolic contents, 'Heguoyihao' showed the highest total and free flavonoids contents, and 'Chushuhong' showed the highest total and cellular antioxidant activities. Eight phenolic compounds were detected, and chlorogenic acid, gallic acid, and quercetin were shown as three predominant components in all five cultivars. These results provide valuable information which may be a guidance for selection of good chestnut variety to be used as functional food.
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Antioxidantes/análise , Antioxidantes/farmacologia , Fagaceae/química , Flavonoides/análise , Fenóis/análise , Compostos Fitoquímicos/análise , Antioxidantes/química , Flavonoides/química , Frutas/química , Células Hep G2 , Humanos , Fenóis/química , Compostos Fitoquímicos/química , Extratos Vegetais/químicaRESUMO
BACKGROUND: Late diagnosis of a salivary fistula increases the risk of wound infection and scarring. The purpose of the present study was to identify a quantitative predictor of postoperative salivary fistula for cases treated with surgery. METHODS: Demographic, intraoperative and postoperative parameters for 57 cases that received surgery for benign parotid tumours were recorded from June 2017 to June 2018, of which 18 cases developed salivary fistulas. These data were analysed using univariate and binary logistic regression analyses as well as receiver operating curve analysis. RESULTS: Drain fluid amylase concentration was positively correlated with salivary fistula development (p < 0.001), with an odds ratio of 1.14 for a 1 KU/L increase in concentration and an optimal receiver operating curve cut-off value of 51,100 U/L for predicting salivary fistula development. Cases wherein the parotid-masseteric fascia remained intact were associated with a lower risk of salivary fistula development (p = 0.006). CONCLUSION: Drain fluid amylase concentration may be a valuable predictor of postoperative salivary fistula in cases with benign parotid tumours.
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Amilases , Glândula Parótida/cirurgia , Neoplasias Parotídeas/cirurgia , Complicações Pós-Operatórias , Fístula das Glândulas Salivares/etiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Amilases/sangue , Drenagem , Humanos , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Otorrinolaringológicos/métodos , Pancreaticoduodenectomia/efeitos adversos , Glândula Parótida/patologia , Valor Preditivo dos Testes , Resultado do Tratamento , Adulto JovemRESUMO
Dual-targeted nanoparticles are gaining increasing importance as a more effective anticancer strategy by attacking double key sites of tumor cells, especially in chemophotodynamic therapy. To retain the nuclei inhibition effect and enhance doxorubicin (DOX)-induced apoptosis by mitochondrial pathways simultaneously, we synthesized the novel nanocarrier (HKH) based on hollow carbon nitride nanosphere (HCNS) modified with hyaluronic acid (HA) and the mitochondrial localizing peptide D[KLAKLAK]2 (KLA). DOX-loaded HKH nanoparticles (HKHDs) showed satisfactory drug-loading efficiency, excellent solubility, and very low hemolytic effect. HA/CD44 binding and electrostatic attraction between positively charged KLA and A549 cells facilitated HKHD uptake via the endocytosis mechanism. Acidic microenvironment, hyaluronidase, and KLA targeting together facilitate doxorubicin toward the mitochondria and nuclei, resulting in apoptosis, DNA intercalation, cell-cycle arrest at the S phase, and light-induced reactive oxygen species production. Intravascular HKHD inhibited tumor growth in A549-implanted mice with good safety. The present study, for the first time, systemically reveals biostability, targetability, chemophotodynamics, and safety of the functionalized novel HKHD.
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Núcleo Celular , Doxorrubicina , Sistemas de Liberação de Medicamentos , Mitocôndrias , Nanosferas , Nitrilas , Fotoquimioterapia , Animais , Feminino , Humanos , Camundongos , Células A549 , Peptídeos Catiônicos Antimicrobianos/química , Apoptose/efeitos dos fármacos , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Doxorrubicina/química , Doxorrubicina/farmacologia , Doxorrubicina/uso terapêutico , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos , Endocitose/efeitos dos fármacos , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/química , Camundongos Endogâmicos BALB C , Camundongos Nus , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Nanosferas/química , Nitrilas/química , Fotoquimioterapia/métodos , Espécies Reativas de Oxigênio/metabolismo , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos , Solubilidade , Distribuição Tecidual , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
A perfect microenvironment facilitates the activated circulating tumor cells (CTCs) to spark the adhesion-invasion-extravasation metastatic cascade in their premetastatic niche. Platelet-CTC interaction contributes to the progression of tumor malignancy by protecting CTCs from shear stress and immunological assault, aiding CTCs entrapment in the capillary bed, enabling CTCs to successfully exit the bloodstream and enter the tissue, inducing epithelial-mesenchymal-like transition (EMT), and assisting in the establishment of metastatic foci. To prevent the cascade from sparking, we show that, the multifunctional S-nitrosocaptopril (CapNO) acts on both CTCs and platelets to interrupt platelet/CTCs interplay and adhesion to endothelium, thus inhibiting CTC-based pulmonary metastasis in vivo. The activated platelets cloak cancer HT29 cells, resulting in HT29-exhibiting platelet biomarkers CD61 and P-selectin positive. CapNO inhibits both sialyl Lewisx (Slex) expression on HT29 and ADP-induced activation of platelets through P-selectin- and GPIIb/IIIa-dependent mechanisms, confirmed by the corresponding antibody assay. CapNO inhibits platelet- or interleukin (IL)-1ß-mediated adhesion between HT29 and endothelial cells, and micrometastatic formation in the lungs of immunocompetent syngeneic mouse models. CapNO have also shown the effects of vasodilation, anticoagulation, inhibition of matrix metalloproteinase-2 (MMP2) expression on cancer cells, and inhibition of cell adhesion molecules (CAMs) expression on vascular endothelium. Due to a series of the beneficial effects of CapNO, CTCs remain exposed to the hostile bloodstream environment and are vulnerable to death induced by shear stress and immune elimination. This new discovery provides a basis for CapNO used for cancer metastatic chemoprevention, and might suggest regulation of the CTCs bloodstream microenvironment as a new avenue for cancer metastatic prevention.
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Antineoplásicos/uso terapêutico , Captopril/análogos & derivados , Neoplasias/tratamento farmacológico , Células Neoplásicas Circulantes/efeitos dos fármacos , Animais , Antineoplásicos/farmacologia , Plaquetas/efeitos dos fármacos , Plaquetas/fisiologia , Captopril/farmacologia , Captopril/uso terapêutico , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Feminino , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Masculino , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Neoplasias/metabolismo , Neoplasias/patologia , Selectina-P/metabolismo , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/metabolismoRESUMO
BACKGROUND: Molybdenum disulfide (MoS2) has been widely explored for biomedical applications due to its brilliant photothermal conversion ability. In this paper, we report a novel multifunctional MoS2-based drug delivery system (MoS2-SS-HA). By decorating MoS2 nanosheets with hyaluronic acid (HA), these functionalized MoS2 nanosheets have been developed as a tumor-targeting chemotherapeutic nanocarrier for near-infrared (NIR) photothermal-triggered drug delivery, facilitating the combination of chemotherapy and photothermal therapy into one system for cancer therapy. RESULTS: The nanocomposites (MoS2-SS-HA) generated a uniform diameter (ca. 125 nm), exhibited great biocompatibility as well as high stability in physiological solutions, and could be loaded with the insoluble anti-cancer drug erlotinib (Er). The release of Er was greatly accelerated under near infrared laser (NIR) irradiation, showing that the composites can be used as responsive systems, with Er release controllable through NIR irradiation. MTT assays and confocal imaging results showed that the MoS2-based nanoplatform could selectively target and kill CD44-positive lung cancer cells, especially drug resistant cells (A549 and H1975). In vivo tumor ablation studies prove a better synergistic therapeutic effect of the joint treatment, compared with either chemotherapy or photothermal therapy alone. CONCLUSION: The functionalized MoS2 nanoplatform developed in this work could be a potent system for targeted drug delivery and synergistic chemo-photothermal cancer therapy.
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Antineoplásicos/farmacologia , Dissulfetos/química , Portadores de Fármacos/química , Cloridrato de Erlotinib/farmacologia , Hipertermia Induzida , Molibdênio/química , Nanocompostos/química , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Liberação Controlada de Fármacos , Cloridrato de Erlotinib/química , Feminino , Humanos , Ácido Hialurônico/química , Concentração de Íons de Hidrogênio , Raios Infravermelhos , Camundongos Endogâmicos BALB C , Camundongos Nus , FototerapiaRESUMO
We report on a novel method for saliva identification by reverse transcription-loop-mediated isothermal amplification (RT-LAMP). In our previous report, real-time RT-LAMP was used for blood identification by using HBB detection as a model but in this advanced study, this method was refined for the identification of the more challenging body fluid of saliva. Expression of the18S rRNA gene was used as the internal control and the Statherin (STATH) gene as the saliva-specific marker. A turbidimeter was used for real-time detection of the RT-LAMP products, and confirmation was obtained that the real products were generated using: agarose gel electrophoresis, calcein fluorescence detection and/or enzymatic digestion. The specificity of the test was performed using 42 samples including 7 different body fluids, and the expression of STATH was only observed in all the saliva samples (6) with a threshold time of 39.4 ± 2.9 min. Sensitivity testing showed that RT-LAMP products for STATH were stably detected when the RNA template was not less than 6.25 ng. When the primer concentrations for STATH were two times that of 18S rRNA, saliva could be identified in the body fluid mixtures even at a ratio (saliva:semen) of 1:3 (without loop primer)/1:5 (with loop primer). A multiplex RT-LAMP was established to simultaneously amplify the 18S rRNA and STATH genes, and applied to the identification of saliva on ten non-probative cigarette butts. A positive result for saliva was obtained from all ten butts, even for those that returned a negative or ambiguous result using the amylase test. A direct RT-LAMP test is also reported where the RNA extraction step was omitted to speed the collection of data and all tests using either the simplex or multiplex RT-LAMP resulted in a positive response if saliva was present. Our data provide a simple and effective means to detect the presence of saliva.
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Técnicas de Amplificação de Ácido Nucleico/métodos , Saliva/química , Amilases/análise , Biomarcadores/análise , Medicina Legal/métodos , Marcadores Genéticos , Humanos , RNA Ribossômico 18S/metabolismo , Proteínas e Peptídeos Salivares/genética , Sensibilidade e EspecificidadeRESUMO
The circulating tumor cells (CTCs) existing in cancer survivors are considered the root cause of cancer metastasis. To prevent the devastating metastasis cascade from initiation, we hypothesize that a biodegradable nanomaterial loaded with the abortifacient mifepristone (MIF) and conjugated with the epithelial cell adhesion molecule antibody (aEpCAM) may serve as a safe and effective cancer metastatic preventive agent by targeting CTCs and preventing their adhesion-invasion to vascular intima. It is demonstrated that MIF-loaded mesoporous silica nanoparticles (MSN) coated with aEpCAM (aE-MSN-M) can specifically target and bind colorectal cancer cells in either cell medium or blood through EpCAM recognition proven by quantitative flow cytometric detection and free aEpCAM competitive assay. The specific binding results in downregulation of the captured cells and drives them into G0/G1 phase primarily attributed to the effect of aEpCAM. The functional nanoparticles significantly inhibit the heteroadhesion between cancer cells and endothelial cells, suggesting the combined inhibition effects of aEpCAM and MIF on E-selectin and ICAM-1 expression. The functionalized nanoparticles circulate in mouse blood long enough to deliver MIF and inhibit lung metastasis. The present proof-of-concept study shows that the aE-MSN-M can prevent cancer metastasis by restraining CTC activity and their adhesion-invasion to vascular intima.
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Anticorpos Monoclonais/imunologia , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/secundário , Molécula de Adesão da Célula Epitelial/imunologia , Mifepristona/administração & dosagem , Nanocápsulas/química , Dióxido de Silício/química , Abortivos Esteroides/administração & dosagem , Abortivos Esteroides/química , Absorção Fisico-Química , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/química , Apoptose/efeitos dos fármacos , Neoplasias Colorretais/imunologia , Difusão , Sistemas de Liberação de Medicamentos/métodos , Reposicionamento de Medicamentos , Células HT29 , Humanos , Camundongos , Mifepristona/química , Nanocápsulas/ultraestrutura , Nanoconjugados/administração & dosagem , Nanoconjugados/química , Nanoporos/ultraestrutura , Resultado do TratamentoRESUMO
BACKGROUND: This study was aimed at establishing a sensitive and specific isolation, characterization, and enumeration method for living circulating tumor cells (CTCs) in patients with colorectal carcinoma. METHODS: Quantitative isolation and characterization of CTCs were performed through a combination of immunomagnetic negative enrichment and fluorescence-activated cell sorting. Isolated CTCs were identified by immunofluorescence staining. The viability and purity of the sorted cells were determined by flow cytometry. Blood samples spiked with HCT116 cells (range, 3-250 cells) were used to determine specificity, recovery, and sensitivity. The method was used to enumerate, characterize, and isolate living CTCs in 10 mL of blood from patients with colorectal carcinoma. RESULTS: The average recovery of HCT116 cells was 61% or more at each spiking level, and the correlation coefficient was 0.992. An analysis of samples from all 18 patients with colorectal carcinoma revealed that 94.4% were positive for CTCs with an average of 33 ± 24 CTCs per 10 mL of blood and with a diameter of 14 to 20 µm (vs 8-12 µm for lymphoma). All patients were CD47(+) , with only 4.3% to 61.2% being CD44(+) . The number of CTCs was well correlated with the patient TNM stage and could be detected in patients at an early cancer stage. The sorted cells could be recultured, and their viability was preserved. CONCLUSIONS: This method provides a novel technique for highly sensitive and specific detection and isolation of CTCs in patients with colorectal carcinoma. This method complements the existing approaches for the de novo functional identification of a wide variety of CTC types. It is likely to help in predicting a patient's disease progression and potentially in selecting the appropriate treatment.
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Neoplasias Colorretais/patologia , Células Neoplásicas Circulantes/patologia , Antígeno CD47/metabolismo , Linhagem Celular Tumoral , Neoplasias Colorretais/metabolismo , Citometria de Fluxo , Humanos , Receptores de Hialuronatos/metabolismo , Separação Imunomagnética , Estadiamento de Neoplasias , Células Neoplásicas Circulantes/metabolismo , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Cancer metastasis caused by circulating tumor cells (CTCs) accounts for 90% cancer-related death worldwide. Blocking the circulation of CTCs in bloodstream and their hetero-adhesion to vascular endothelia of the distant metastatic organs may prevent cancer metastasis. Nanomaterial-based intervention with adhesion between CTCs and endothelia has not been reported. Driven by the novel idea that multivalent conjugation of EpCAM and Slex antibodies to dendrimer surface may enhance the capacity and specificity of the nanomaterial conjugates for capturing and down-regulating colorectal CTCs, we conjugated the dendrimer nanomaterial with the EpCAM and Slex antibodies, and examined the capacity of the dual antibody-coated nanomaterial for their roles in interrupting CTCs-related cancer metastasis. RESULTS: The antibody-coated nanomaterial was synthesized and characterized. The conjugates specifically bound and captured colon cancer cells SW620. The conjugate inhibited the cells' viability and their adhesion to fibronectin (Fn)-coated substrate or human umbilical vein endothelial cells (HUVECs) in a concentration-dependent manner. In comparison with SW480 and LoVo cell lines, the activity and adhesion of SW620 to Fn-coated substrate and HUVECs were more specifically inhibited by the dual antibody conjugate because of the higher levels of EpCAM and Slex on SW620 cell surface. The hetero-adhesion between SW620 and Fn-coated substrate, or HUVECs was inhibited by about 60-70%. The dual conjugate showed the inhibition capacity more significant than its corresponding single antibody conjugates. CONCLUSIONS: The present study provides the new evidence that coating nanomaterials with more than one antibody against CTCs may effectively interfere with the interaction between SW620 and HUVECs.
Assuntos
Antineoplásicos/farmacologia , Adesão Celular/efeitos dos fármacos , Endotélio Vascular/citologia , Nanoestruturas , Células Neoplásicas Circulantes/efeitos dos fármacos , Anticorpos/química , Antígenos de Neoplasias/imunologia , Antineoplásicos/química , Moléculas de Adesão Celular/imunologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Proliferação de Células , Dendrímeros/química , Relação Dose-Resposta a Droga , Molécula de Adesão da Célula Epitelial , Fibronectinas/química , Fibronectinas/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nanoestruturas/química , Metástase Neoplásica/prevenção & controle , Células Neoplásicas Circulantes/metabolismo , Células Neoplásicas Circulantes/patologiaRESUMO
Glycolate oxidase (GLO) is a key enzyme for photorespiration in plants. Previous studies have demonstrated that suppression of GLO causes photosynthetic inhibition, and the accumulated glycolate with the deactivated Rubisco is likely involved in the regulation. Using isolated Rubisco and chloroplasts, it has been found that only glyoxylate can effectively inactivate Rubisco and meanwhile inhibit photosynthesis, but little in vivo evidence has been acquired and reported. In this study, we have generated the transgenic rice (Oryza sativa) plants with GLO being constitutively silenced, and conducted the physiological and biochemical analyses on these plants to explore the regulatory mechanism. When GLO was downregulated, the net photosynthetic rate (Pn) was reduced and the plant growth was correspondingly stunted. Surprisingly, glyoxylate, as a product of the GLO catalysis, was accumulated in response to the GLO suppression, like its substrate glycolate. Furthermore, the glyoxylate content was found to be inversely proportional to the Pn while the Pn is directly proportional to the Rubisco activation state in the GLO-suppressed plants. A mathematical fitting equation using least square method also demonstrated that the Rubisco activation state was inversely proportional to the glyoxylate content. Despite that the further analyses we have conducted failed to reveal how glyoxylate was accumulated in response to the GLO suppression, the current results do strongly suggest that there may exist an unidentified, alternative pathway to produce glyoxylate, and that the accumulated glyoxylate inhibits photosynthesis by deactivating Rubisco, and causes the photorespiratory phenotype in the GLO-suppressed rice plants.
Assuntos
Oxirredutases do Álcool/metabolismo , Glicolatos/metabolismo , Oryza/metabolismo , Fotossíntese , Proteínas de Plantas/metabolismo , Ribulose-Bifosfato Carboxilase/metabolismo , Oxirredutases do Álcool/genética , Western Blotting , Cloroplastos/genética , Cloroplastos/metabolismo , Inativação Gênica , Oryza/genética , Fenótipo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ribulose-Bifosfato Carboxilase/genética , Transdução de Sinais/genéticaRESUMO
CoCrNi medium-entropy alloys (MEAs) have attracted extensive attention and research because of their superior mechanical properties, such as higher ductility, strength, and toughness. This study uses molecular dynamics (MD) simulations to investigate the cutting behavior of a gradient nanograined (GNG) CoCrNi MEA. Moreover, it explores the influence of relative tool sharpness and rake angle on the cutting process. The results show that an increase in the average grain size of the GNG samples leads to a decrease in the average resultant cutting force, as predicted by the Hall-Petch relationship. The deformation behavior shows that grain boundaries are crucial in inhibiting the propagation of strain and stress. As the average grain size of the GNG sample increases, the range of shear strain distribution and average von Mises stress decreases. Moreover, the cutting chips become thinner and longer. The subsurface damage is limited to a shallow layer at the surface. Since thermal energy is generated in the high grain boundary density, the temperature of the contact zone between the substrate and the cutting tool increases as the GNG size decreases. The cutting chips removed from the GNG CoCrNi MEA substrates will transform into a mixed structure of face-centered cubic and hexagonally close-packed phases. The sliding and twisting of grain boundaries and the merging of grains are essential mechanisms for polycrystalline deformation. Regarding the cutting parameters, the average resultant force, the material accumulation, and the chip volume increase significantly with the increase in cutting depth. In contrast to sharp tools, which mainly use shear deformation, blunt tools remove material by plowing, and the cutting force increases with the increase in cutting-edge radius and negative rake angle.
RESUMO
Liquid biopsies utilizing tumor exosomes offer a noninvasive approach for cancer diagnosis. However, validation studies consistently report that in the early stages of cancer, the secretion of exosomes by cancer cells is relatively low, while bodily fluids exhibit a high abundance of other interfering biomolecules. Additionally, target mutations or differences in biomarker expression among various lung cancer subtypes may contribute to detection failures. In this study, we propose a targeted nanoarray-based early cancer diagnostic approach for multiple subtypes of lung cancer. The targeted nanoarray was constructed by modifying five targeting aptamers onto mesoporous silica nanoparticles through the conjugation between amino and carboxyl groups. The flow cytometry experiments demonstrated the specific recognition ability of the targeted nanoarray to tumor exosomes in PBS, even at biomarker expression levels as low as 1.5 %. Moreover, the TEM results indicated that the targeted nanoarray could isolate tumor exosomes in the blood of tumor-bearing mice. Furthermore, the targeted nanoarray could detect tumor exosomes in the blood of various lung cancer bearing mice, including at the early stages of cancer, which has just been established for 7 days. Overall, the targeted nanoarray represents a promising tool for the early detection of various subtypes of lung cancer.