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BACKGROUND: In response to growing concerns regarding heavy metal contamination in food, particularly chromium (Cr)(VI) contamination, this study presented a simple, sensitive and practical method for Cr(VI) detection. RESULTS: A magnetic separation-based capture-exponential enrichment ligand system evolution (SELEX) method was used to identify and characterize DNA aptamers with a high affinity for Cr(VI). An aptamer, Cr-15, with a dissociation constant (Kd) of 4.42 ± 0.44 µmol L-1 was obtained after only eight rounds of selection. Further innovative methods combining molecular docking, dynamic simulation and thermodynamic analysis revealed that CrO4 2- could bind to the 19th and 20th guanine bases of Cr-15 via hydrogen bonds. Crucially, a label-free fluorometric aptasensor based on SYBR Green I was successfully constructed to detect CrO4 2-, achieving a linear detection range of 60-300 nmol L-1 with a lower limit of detection of 44.31 nmol L-1. Additionally, this aptasensor was able to quantitatively detect CrO4 2- in grapes and broccoli within 40 min, with spike recovery rates ranging from 89.22% to 108.05%. The designed fluorometric aptasensor exhibited high selectivity and could detect CrO4 2- in real samples without sample processing or target pre-enrichment. CONCLUSION: The aptasensor demonstrated its potential as a reliable tool for monitoring Cr(VI) contamination in fruit and vegetable products. © 2024 Society of Chemical Industry.
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Aptamers are functional single-stranded oligonucleotide fragments isolated from randomized libraries by Systematic Evolution of Ligands by Exponential Enrichment (SELEX), exhibiting excellent affinity and specificity toward targets. Compared with traditional antibody reagents, aptamers display many desirable properties, such as low variation and high flexibility, and they are suitable for artificial and large-scale synthesis. These advantages make aptamers have a broad application potential ranging from biosensors, bioimaging to therapeutics and other areas of application. However, the overall performance of aptamer pre-selected by SELEX screening is far from being satisfactory. To improve aptamer performance and applicability, various post-SELEX optimization methods have been developed in the last decade. In this review, we first discuss the key factors that influence the performance or properties of aptamers, and then we summarize the key strategies of post-SELEX optimization which have been successfully used to improve aptamer performance, such as truncation, extension, mutagenesis and modification, splitting, and multivalent integration. This review shall provide a comprehensive summary and discussion of post-SELEX optimization methods developed in recent years. Moreover, by discussing the mechanism of each approach, we highlight the importance of choosing the proper method to perform post-SELEX optimization.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Técnica SELEX de Producción de Aptámeros/métodos , Ligandos , AnticuerposRESUMEN
Black tea is one of the six major tea categories and has a variety of bioactivities. However, little is known about its comprehensive evaluation of hypoglycemic effects and potential mechanisms. In this study, we investigated the in vivo hypoglycemic activity and potential mechanism for aqueous extracts of ordinary black tea (BT) and selenium-enriched black tea (Se-BT) by using an established high-fat diet together with streptozotocin (STZ)-induced hyperglycemic mouse model. Additionally, we also explored their α-glucosidase inhibition activity. The results show that both BT and Se-BT had a favorable glycosidase inhibitory activity. Moreover, the intervention of BT and Se-BT could regulate the mRNA expression and the level of serum parameters related to glucose and lipid metabolisms. Accordingly, they could activate the phosphoinositide-3-kinase/protein kinase B (PI3K/Akt) signaling pathway and alleviate insulin resistance (IR) and hyperglycemia. Moreover, supplementation of BT and Se-BT increased the richness and diversity of intestinal flora and altered the abundance of beneficial and harmful bacteria. Both BT and Se-BT could regulate glucose metabolism, alleviate tissue damage, and restore intestinal flora dysbiosis, suggesting that they could be used as a natural functional food for preventing hyperglycemia.
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Camellia sinensis , Microbioma Gastrointestinal , Hiperglucemia , Selenio , Animales , Ratones , Glucosa , Té , Ratones Obesos , Glucemia/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Hipoglucemiantes/farmacología , Camellia sinensis/metabolismo , LípidosRESUMEN
Various functional components in tea have been well developed, but less research has been explored on glycoproteins in tea. In this paper, three types of glycoprotein fractions, namely tea selenium-binding glycoprotein1-1 (TSBGP1-1), TSBGP2-1, and TSBGP3-1, respectively, were extracted and purified from selenium-enriched coarse green tea. Chemical analysis revealed that three fractions were glycoproteins, but their selenium content, molecular weight, and monosaccharide composition were significantly different. Fourier transforms infrared (FT-IR) analysis indicated that three fractions contained characteristic absorption peaks of glycoproteins but differed in secondary structural composition. Thermogravimetric (TG) analysis showed that the thermal stability of the three fractions was dramatically distinct. The in vitro hypoglycemic activity showed that TSBGPs significantly activated the insulin receptor substrate 2 (IRS2)/protein kinase B (Akt) pathway in LO2 cells, then enhanced glucose metabolism and inhibited gluconeogenesis, and finally ameliorated insulin resistance (IR) and glucose metabolism disorders. Furthermore, Pearson correlation analysis reveals that the hypoglycemic activity was significantly correlated with Se, protein, monosaccharide composition (especially glucose), molecular weight, and secondary structure. Our results show that Se-enriched tea glycoprotein is a desirable candidate for developing anti-diabetic food, and TSBGP-2 and TSBGP-3 had a better regulation effect. Our results can provide a research reference for the extraction, physicochemical property, and function of selenium-enriched plant glycoproteins.
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Selenio , Glicoproteínas , Hipoglucemiantes/análisis , Hipoglucemiantes/farmacología , Monosacáridos/análisis , Selenio/análisis , Espectroscopía Infrarroja por Transformada de Fourier , Té/químicaRESUMEN
BACKGROUND: Dark tea, comprising one of the six major teas, has many biological activities, which originate from their active substrates, such as polyphenols, polysaccharides, and so on. The hypoglycemic effect is one of its most prominent activities, although less is known about their evaluation and potential role in the hypoglycemic mechanism. RESULTS: In the present study, we separately analyzed the phytochemical composition, glycosidase inhibition and free radical scavenging activities, and hypoglycemic activity in type 2 diabetes mellitus mice, as well as the alleviation of insulin resistance in HepG2 cells of four dark tea aqueous extracts. The results showed that the phytochemical composition of dark tea aqueous extracts was significantly different, and they all had good glycosidase inhibition and free radical scavenging activities, in vivo hypoglycemic activity and alleviation of insulin resistance, and could also activate the phosphatidylinositol 3-kinase-Akt-perixisome proliferation-activated receptor cascade signaling pathway to regulate glucose and lipid metabolism, change the key enzyme activities related to glucose metabolism and antioxidant activity, and reduce oxidative stress and inflammatory factor levels. Among them, Liubao brick tea (LBT) and Pu-erh tea (PET) possessed better glycosidase inhibitory activity, in vivo hypoglycemic activity and improved insulin resistance activity, whereas Qingzhuan brick tea and Fuzhuan brick tea had better free radical scavenging activity, which may be explained by their distinct phytochemical compositions, such as tea proteins, polysaccharides, polyphenols, catechins, and tea pigments and some elements. CONCLUSION: Dark tea is a highly attractive candidate for developing antidiabetic food, LBT and PET may be good natural sources of agricultural products with anti-diabetic effects. © 2021 Society of Chemical Industry.
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Glucemia/metabolismo , Camellia sinensis/metabolismo , Diabetes Mellitus Tipo 2/dietoterapia , Hipoglucemiantes/metabolismo , Resistencia a la Insulina , Hígado/metabolismo , Fitoquímicos/metabolismo , Animales , Antioxidantes/química , Antioxidantes/metabolismo , Camellia sinensis/química , China , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/metabolismo , Glicósido Hidrolasas/genética , Glicósido Hidrolasas/metabolismo , Células Hep G2 , Humanos , Hipoglucemiantes/química , Masculino , Ratones , Ratones Endogámicos C57BL , Estrés Oxidativo , Receptores Activados del Proliferador del Peroxisoma/genética , Receptores Activados del Proliferador del Peroxisoma/metabolismo , Fitoquímicos/química , Extractos Vegetales/química , Extractos Vegetales/metabolismo , TéRESUMEN
Coarse tea is made of mature tea plant (Camellia sinensis L.) shoots and is generally discarded as a worthless crop product, but has been proved an excellent material for the treatment of diabetes. This study aims to evaluate the effects of the extraction techniques WE (water extraction), UAE (ultrasound-assisted extraction), MAE (microwave-assisted extraction), and EE (enzyme extraction) on the physicochemical properties and antidiabetic activities of polysaccharides from coarse tea (CTPSs). The results showed that all four CTPSs had homogeneity in the monosaccharide types and similar IR (Infrared spectroscopy) characteristic absorption peaks, but differed in monosaccharide proportion and molecular weight distribution. Compared with the other three extraction techniques, CCTPS extracted by EE had the lowest protein content, the highest total sugar content of 71.83% and a polysaccharide yield of 4.52%. In addition, EE-CTPS had the best hypoglycemic activity that was better than ordinary green tea polysaccharides, the α-glucosidase and α-amylase inhibitory activities of EE-CTPS were highest in the range of 2-10 mg/mL compared with the other three CTPSs, which may be related to its smaller molecular weight and porous structure. The results suggested that the EE method was a good way to extract polysaccharides from coarse tea for food and pharmaceutical production.
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Fraccionamiento Químico/métodos , Inhibidores Enzimáticos/análisis , Hipoglucemiantes/análisis , Polisacáridos/análisis , Té/química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Glucosidasas/antagonistas & inhibidores , Hipoglucemiantes/química , Hipoglucemiantes/farmacología , Microondas , Polisacáridos/química , Polisacáridos/farmacología , Ondas Ultrasónicas , alfa-Amilasas/antagonistas & inhibidoresRESUMEN
Pancreatic lipase (PL) and cholesterol esterase (CE) are vital digestive enzymes that regulate lipid digestion. Three bioactive peptides (LFCMH, RIPAGSPF, YFRPR), possessing enzyme inhibitory activities, were identified in the seed proteins of R. roxburghii. It is hypothesized that these peptides could inhibit the activities of these enzymes by binding to their active sites or altering their conformation. The results showed that LFCMH exhibited superior inhibitory activity against these enzymes compared to the other peptides. The inhibition mechanisms of the three peptides were identified as either competitive or mixed, according to inhibition models. Further studies have shown that peptides could bind to the active sites of enzymes, thus affecting their spatial conformation and restricting substrate entry into the active site. Molecular simulation further proved that hydrogen bonds and hydrophobic interactions played a vital role in the binding of peptides to enzymes. This study enriches our understanding of interaction mechanisms of peptides on PL and CE.
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Inhibidores Enzimáticos , Esterol Esterasa , Inhibidores Enzimáticos/farmacología , Lipasa/química , Péptidos/farmacología , TermodinámicaRESUMEN
Cadmium (Cd2+) is a highly toxic heavy metal that can accumulate in the human body through contaminated food and water, posing great health risks. In this study, a label-free fluorescent aptasensor based on SYBR Green I (SGI) for the rapid and sensitive detection of Cd2+ in food samples was designed. The aptasensor utilizes a Cd2+-specific aptamer (Cd-(21)) and its complementary strand (CSCd-(21)) to form a double-stranded DNA (dsDNA) structure in the absence of Cd2+. SGI intercalates into the dsDNA, resulting in a strong fluorescence signal. In the presence of Cd2+, the aptamer undergoes a conformational change, preventing the formation of dsDNA and leading to a decrease in fluorescence intensity. Under optimized conditions, the aptasensor exhibited a linear response to Cd2+ concentrations ranging from 0.11 to 157.37 ng mL-1, with a limit of detection (LOD) of 0.07 ng mL-1. The aptasensor demonstrated high specificity and was successfully applied to detect Cd2+ in fruits and vegetables, with satisfactory recovery rates (95-111%). The proposed aptasensor provides a promising tool for the rapid and sensitive detection of Cd2+ in food.
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Fermented foods have shown promise in preventing or treating ulcerative colitis (UC) via regulating intestinal flora and correcting metabolic disorders. However, the prevention effect of fermented Wallace melon juice (FMJ) on UC is unclear. In this study, the effects of FMJ on dextran sodium sulfate (DSS)-induced UC were investigated via 16S rRNA sequencing and non-targeted metabolomics. The results showed that FMJ was effective in alleviating the symptoms of UC, reducing histological damage and oxidative stress, decreasing the levels of pro-inflammatory cytokines. After FMJ treatment, the level of propionic acid, butyric acid, and valeric acid increased by 14.1%, 44.4%, and 52.4% compared to DSS-induced UC mice. Meanwhile, the levels of harmful bacteria such as Oscillospira, Bacteroidetes, and Erysipelotrichaceae and Clostridium decreased, while the levels of beneficial bacteria such as Akkermansia, Lactobacillus, and Bifidobacterium increased. Fecal metabolomics analysis identified 31 differential metabolites, which could regulate metabolic disorders in UC mice by controlling the primary bile acid biosynthesis, purine metabolism, and pantothenate and CoA biosynthesis pathway. Additionally, the abundances of butyric acid, bile acids, and pantothenic acid were positively correlated with Allobaculum, Bifidobacterium, and other beneficial bacteria (R2 > 0.80, p < 0.01). The results indicated that FMJ played a role in regulating the structure of intestinal flora, which in turn helped in repairing metabolic disorders and alleviated colitis inflammation.
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Colitis Ulcerosa , Colitis , Microbioma Gastrointestinal , Enfermedades Metabólicas , Animales , Ratones , Lactobacillus , Colitis Ulcerosa/inducido químicamente , Sulfato de Dextran/efectos adversos , ARN Ribosómico 16S , Ácido Butírico , Bifidobacterium , Firmicutes , Ratones Endogámicos C57BL , Modelos Animales de Enfermedad , ColonRESUMEN
A high-sugar and -fat diet (HSFD) has become a primary risk factor for diabetes, and dietary intervention shows a substantial effect on the prevention and management of hyperglycemia. In this study, the chemical compositions of the aqueous extracts of stir-fried green tea (GT) and congou black tea (BT) were compared. Moreover, their potential mechanisms and regulatory effects on hepatic glycolipid metabolism and gut microbiota disorders in hyperglycemic mice were further explored. Our results show that GT or BT intervention had a prominent regulatory effect on glycolipid metabolism. Moreover, they could significantly regulate the levels of serum metabolic signatures, the activities of key enzymes in liver glucose metabolism, and the expression of genes or proteins related to glycolipid metabolism via activating the IRS-1-PI3K/AKT-GLUT2 signaling pathway. Significantly, GT or BT administration adjusted the composition and diversity of the gut microbiota, mainly reflecting a significant increase in the abundance of beneficial bacteria (including Allobaculum, Lactobacillus, and Turicibacter) and reducing the abundance of harmful or conditionally pathogenic bacteria (mainly including Clostridiales and Bacteroides). Our results suggest that dietary supplementation with GT or BT could exert a practical anti-diabetic effect. Meanwhile, BT intervention showed a better regulation effect on glycolipid metabolism. This study reveals that GT and BT have excellent potential for developing anti-diabetic food.
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Camellia sinensis , Microbioma Gastrointestinal , Ratones , Animales , Té/química , Ratones Obesos , Fosfatidilinositol 3-Quinasas , Camellia sinensis/química , Dieta Alta en Grasa/efectos adversos , Glucolípidos/farmacología , Ratones Endogámicos C57BLRESUMEN
Heavy metal pollution has become a growing concern in industrial, agricultural, and manufacturing processes, posing a significant threat to human health. Among these heavy metals, arsenic (As) is highly toxic and shares similar chemical properties and environmental behavior with other heavy metals. As(III) is particularly toxic compared to other forms of arsenic. Therefore, it is essential to develop a real-time, rapid, and sensitive method for the determination of As(III). In this study, we employed a unique bifunctional chelator, 1-(4-isothiocyanobenzyl)-ethylenediamine N,N,N',N'-tetraacetic acid (ITCBE), to prepare a complete antigen. Through a series of tests including balb/c mouse immunization, cell fusion (mouse L2041 spleen cells with mouse myeloma cells SP2/0), and subcloning, we generated four monoclonal cell lines (1C1, 2C2, 3A9, and 4A11). These cell lines demonstrated high purity, high affinity, and IC50 values of less than 50 µg mL-1. Monoclonal antibody 4A11, which exhibited a strong Raman signal, was selected as the probe, and Au@Ag 200 was utilized as the surface-enhanced Raman scattering (SERS) substrate for the preliminary establishment of SERS immunochromatographic test strips. The sensitivity of the SERS immunochromatographic test strips, measured through Raman signal detection, showed a significant improvement compared to the SERS immunochromatographic test strips analyzed by colorimetry (LOD = 49.43 µg mL-1 and LDR = 5.32-81.31 µg mL-1). The SERS immunochromatographic test strips achieved a LOD of 7.62 µg mL-1 and an LDR of 12.66-71.84 µg mL-1. This study presents innovative methodologies for the rapid detection of As(III) using SERS immunochromatographic test strips.
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Arsénico , Nanotubos , Animales , Ratones , Anticuerpos Monoclonales , Oro/química , Inmunoensayo/métodos , Plata/químicaRESUMEN
The research on the activity of selenium (Se)-enriched agricultural products is receiving increasing attention since Se was recognized for its antioxidant activities and for its enhancement of immunity in trace elements. In this study, antioxidant Se-containing peptides, namely, Se-TAPepI-1 and Se-TAPepI-2, were optimally separated and prepared from Se-enriched tea protein hydrolysates by ultrafiltration and Sephadex G-25 purification, and subsequently, their physicochemical properties, oligopeptide sequence, and potential antioxidant mechanism were analyzed. Through the optimization of enzymatic hydrolysis conditions, the Se-enriched tea protein hydrolyzed by papain exhibited a better free radical scavenging activity. After separation and purification of hydrolysates, the two peptide fractions obtained showed significant differences in selenium content, amino acid composition, apparent morphology, peptide sequence, and free radical scavenging activity. Therein, two peptides from Se-TAPepI-1 included LPMFG (563.27 Da) and YPQSFIR (909.47 Da), and three peptides from Se-TAPepI-2 included GVNVPYK (775.42 Da), KGGPGG (552.24 Da), and GDEPPIVK (853.45 Da). Se-TAPepI-1 and Se-TAPepI-2 could ameliorate the cell peroxidation damage and inflammation by regulating NRF2/ARE pathway expression. Comparably, Se-TAPepI-1 showed a better regulatory effect than Se-TAPepI-2 due to their higher Se content, typical amino acid composition and sequence, higher surface roughness, and a looser arrangement in their apparent morphology. These results expanded the functional activities of tea peptide and provided the theoretical basis for the development of Se-containing peptides from Se-enriched tea as a potential natural source of antioxidant dietary supplements.
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Heavy metal ion contamination poses significant environmental and health risks, necessitating rapid and efficient detection methods. In the last decade, colorimetric aptasensors have emerged as powerful tools for heavy metal ion detection, owing to their notable attributes such as high specificity, facile synthesis, adaptability to modifications, long-term stability, and heightened sensitivity. This comprehensive overview summarizes the key developments in this field over the past ten years. It discusses the principles, design strategies, and innovative techniques employed in colorimetric aptasensors using nanomaterials. Recent advancements in enhancing sensitivity, selectivity, and on-site applicability are highlighted. The review also presents application studies of successful heavy metal ion detection using colorimetric aptasensors, underlining their potential for environmental monitoring and health protection. Finally, future directions and challenges in the continued evolution of these aptasensors are outlined.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Metales Pesados , Nanoestructuras , Colorimetría/métodos , Técnicas Biosensibles/métodosRESUMEN
Green tea (GT) and oolong tea (OLT) are widely consumed beverages, and their preventive and regulatory effects on hypertension have been reported. However, the interventional effects of GT and OLT on hypertension induced by a high-salt diet and its mechanism have not been fully explored. This study evaluated the anti-hypertensive effects of GT and OLT and their underlying mechanisms. The in vivo anti-hypertensive effects of GT and OLT and their capability to prevent hypertension and regulate the intestinal microbiota in Wistar rats fed with a high-salt diet were evaluated. Our results show that GT and OLT supplementations could regulate oxidative stress, inflammation, gene expression, and parameter levels related to blood pressure (BP) and prevent the increase in BP induced by a high-salt diet. Furthermore, both GT and OLT boosted the richness and diversity of intestinal microbiota, increased the abundance of beneficial bacteria and reduced the abundance of harmful bacteria and conditionally pathogenic bacteria, and regulated the intestinal microbial metabolism pathway related to BP. Among them, OLT presented better effects than GT. These findings indicate that GT and OLT can prevent hypertension caused by high-salt diets, which may be due to the regulation of intestinal flora by GT and OLT.
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Netilmicin (NET) is an antibiotic widely used in healthcare and agriculture, but it can accumulate in the environment to threat human health. Netilmicin (NET) is an antibiotic used for veterinary purposes, for human therapy and for agricultural purposes. Therefore, there is a need to develop high-sensitive measuring methods to detect NET. Aptamer-based detecting methods are highly sensitive, inexpensive, and portable. In this study, we developed an aptamer-based fluorescence method to detect and quantify NET. NET was first conjugated to magnetic beads by amidation reaction and then NET-coated beads were used as the stationary phase to isolate aptamers by systematic evolution of ligands by exponential enrichment (SELEX) screening method. After ten rounds of SELEX screening, 32 aptamers with NET-binding affinity were obtained and the candidate aptamer APT-21 was finally chosen by comprehensively comparing their secondary structure characters and NET-binding affinity. APT-21 bound to NET with high affinity (Kd = 194.1 nmol/L) and high specificity that it displayed low cross-binding activities on 7 different structural analogs. We also developed a fluorometric assay using SYBR Green I (SG-I) and the APT-21. Key experimental parameters were optimized, including buffer system, SG-I and APT-21 reaction time, SG-I concentration, and aptamer concentration, to improve the detecting sensitivity. Our results suggest that the low limit of detection (LOD) of this method reached a low level of 1.95 nM and it also exhibited a good linear range up to 200 nM. Moreover, we successfully applied our method to detect the NET spiked in tap water and river water with good recoveries in the range from 97% to 111%. In conclusion, our current study isolated a NET-specific aptamer and developed an aptamer-based quantification method, which is promising to apply to detect NET in environmental samples.
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Aptámeros de Nucleótidos , Antibacterianos/análisis , Aptámeros de Nucleótidos/química , Dimaprit/análogos & derivados , Humanos , Netilmicina , Técnica SELEX de Producción de Aptámeros/métodos , AguaRESUMEN
Selenium-enriched mung bean (Se-MB) is a combination of mung bean (MB) and selenium (Se), which have a variety of potential biological activities. However, little is known about the skincare activity of Se-MB. The chemical composition of Se-MB fermentation broth (Se-MBFB) was analyzed to investigate the whitening, moisturizing, and anti-aging activities of Se-MBFB. The tyrosinase inhibition, anti-melanogenic in melanocytes (B16F10 cells), and moisturizing effect in human dermal fibroblasts (HDFs) were analyzed. Besides, the free radical scavenging activity of Se-MBFB was assessed in vitro. To verify the in vivo effects and the potential of practical applications of Se-MBFB, a clinical trial was conducted on the participants: 31 Chinese women aged 25-60 years, with no pigmentation disorder, no illness, no history of hypersensitivity reaction, and no use of skincare product on the face. The participants used an Se-MBFB masque for 15-20 min after cleaning the face. The measurement points were Week 0, 2, and 4 (W0, W2, and W4) after using the masque, and target sites were cheek and canthus. The following parameters were recorded on the target sites at each visit: melanin index, skin color, cuticle moisture content, transepidermal water loss, and crow's feet. The results demonstrated that Se-MBFB was rich in polyphenols, peptides, and γ-aminobutyric acid (GABA), displayed significant free radical scavenging and tyrosinase inhibiting activities, decreased the synthesis of melanin, and upregulated the aquaporin-3 (AQP3) expression. The test of the Se-MBFB mask showed that after 4 weeks of using the Se-MBFB facemask, the faces of the participants became whiter with reduced wrinkles and increased moisture content. Se-MB possessed the excellent whitening, moisturizing, and antioxidant efficacy, which could lay a scientific foundation for utilization and development of skincare products of Se-MB and its related industrial cosmetics products.
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Diabetes and its related metabolic disorders are worldwide public health issues. Many studies have shown that changes in the structure and composition of the intestinal flora are closely related to the host's physiological and pathological processes. In this study, we aim to explore the effect of Liubao tea (LBT) extract on hyperglycemic mice with metabolic disorders and intestinal flora dysbiosis and to further study its regulatory effect on insulin resistance and its potential regulatory mechanism. Our results show that LBT had a good hypoglycemic effect and could significantly alleviate the metabolic disorder evoked by hyperglycemia. The gut microbial sequencing showed that LBT treatment increased the diversity of intestinal flora, increased the abundance of beneficial bacteria, and reduced the abundance of harmful or conditional pathogenic bacteria, as well as significantly altered 39 of the top 50 OTUs with abundance. Besides, LBT could activate the PI3K-Akt-PPARs-GLUT2 cascade signaling pathway to improve metabolic disorders, thereby alleviating insulin resistance. These results suggest that LBT has excellent potential to become a natural functional food for the prevention of hyperglycemia and insulin resistance.
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Microbioma Gastrointestinal , Resistencia a la Insulina , Enfermedades Metabólicas , Animales , Glucemia , Enfermedades Metabólicas/tratamiento farmacológico , Ratones , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , TéRESUMEN
Recently, concerns about heavy metal cadmium ion (Cd2+) residue in asparagus have been frequently reported, and there is an urgent need to develop an effective, sensitive, and rapid detection method for Cd2+. In this study, we innovatively combined molecular microbiology to carry out the comparative screening of Cd2+ chelators in a green, efficient, and specific way. The knock-out putative copper-transporter gene (pca1Δ) yeast strain with high sensitivity to Cd2+ was first used to screen the Cd2+ chelator, and the optimum chelator 1-(4-Isothiocyanatobenzyl)ethylenediamine-N,N,N,N'-tetraacetic acid (ITCBE) was obtained. Additionally, a rapid latex microsphere immunochromatographic assay (LMIA) was developed, based on the obtained monoclonal antibody (mAb) with high specificity and high affinity (affinity constant Ka = 1.83 × 1010 L/mol), to detect Cd2+ in asparagus. The 50% inhibitive concentration (IC50) of test strip was measured to be 0.2 ng/mL, and the limit of detection (IC10) for qualitative (LOD, for visual observation) and quantitative detection (LOQ, for data simulation) of the test strip was 2 ng/mL and 0.054 ng/mL, respectively. In all, the developed mAb-based LMIA shows a great potential for monitoring Cd2+ in asparagus, even in vegetable samples.
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The structure and hypoglycemic activity of tea polysaccharides has been extensively studied, while there are few reports on the characterization and hypoglycemic activity of dark tea polysaccharides. The crude dark tea polysaccharide (CDTPS) was optimally extracted from Fuzhuan dark tea. Six polysaccharide fractions (namely DTPS-1, DTPS-2, DTPS-3, DTPS-4, DTPS-5, and DTPS-6) were isolated from CDTPS, and their physicochemical, structural, and biological properties were compared and analyzed. The results revealed that the compositions, structural characteristics, and biological properties of the six DTPSs were different. Therein, DTPS-4 and DTPS-6 had looser morphology, faster solubility, and a more stable structure. Additionally, DTPS-4 had the optimum in vitro antioxidant capabilities, and DTPS-6 had the strongest in vitro hypoglycemic capabilities. In addition, a correlation analysis revealed that the molecular weight and uronic acid content were significantly related to their antioxidant and hypoglycemic activities. Our results indicated that DTPS-4 and DTPS-6 could be further developed into functional foods or additives, respectively.
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Qingzhuan tea (QZT) with longer aging year is usually believed to have higher quality and commercial value. In this study, a 20 years sequence of aged QZT were subjected to an electronic tongue and liquid chromatography-mass spectrometry to investigate the effect of storage age on its metabolic profile and taste quality. The changes in both taste quality and metabolic profile exhibited a parabolic trend in the 20 years of QZT aging and reached the maximum at the 10th year. A total of 47 compounds were identified as critical metabolites responsible for the age variation of QZT quality, with the methylation of catechins, glycosylation of flavonoids, degradation of flavoalkaloids, biosynthesis of triterpenoids, and formation of theabrownins. These results suggested that the taste of QZT was improved after 10 years of storage, with the reduction of bitterness and astringency and a general increase of key quality-related compounds.