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1.
J Therm Biol ; 119: 103752, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38194751

RESUMEN

Heat stress can lead to hormonal imbalances, weakened immune system, increased metabolic pressure on the liver, and ultimately higher animal mortality rates. This not only seriously impairs the welfare status of animals, but also causes significant economic losses to the livestock industry. Due to its rich residual bioactive components and good safety characteristics, traditional Chinese medicine (TCM) residue is expected to become a high-quality feed additive with anti-oxidative stress alleviating function. This study focuses on the potential of Shengxuebao mixture herbal residue (SXBR) as an anti-heat stress feed additive. Through the UPLC (ultra performance liquid chromatography) technology, the average residue rate of main active ingredients from SXBR were found to be 25.39%. SXBR were then added into the basal diet of heat stressed New Zealand rabbits at the rates of 5% (SXBRl), 10% (SXBRm) and 20% (SXBRh). Heat stress significantly decreased the weight gain, as well as increased neck and ear temperature, drip loss in meat, inflammation and oxidative stress. Also, the hormone levels were disrupted, with a significant increase in serum levels of CA, COR and INS. After the consumption of SXBR in the basal diet for 3 weeks, the weight of New Zealand rabbits increased significantly, and the SXBRh group restored the redness value of the meat to a similar level as the control group. Furthermore, the serum levels T3 thyroid hormone in the SXBRh group and T4 thyroid hormone in the SXBRm group increased significantly, the SXBRh group showed a significant restoration in inflammation markers (IL-1ß, IL-6, and TNF-α) and oxidative stress markers (total antioxidant capacity, HSP-70, MDA, and ROS) levels. Moreover, the real-time fluorescence quantitative PCR analysis found that, the expression levels of antioxidant genes such as Nrf2, HO-1, NQO1, and GPX1 were significantly upregulated in the SXBRh group, and the expression level of the Keap1 gene was significantly downregulated. Additionally, the SXBRm group showed significant upregulation in the expression levels of HO-1 and NQO1 genes. Western blot experiments further confirmed the up-regulation of Nrf2, Ho-1 and NQO1 proteins. This study provides a strategy for the utilization of SXBR and is of great significance for the green recycling of the TCM residues, improving the development of animal husbandry and animal welfare.


Asunto(s)
Antioxidantes , Trastornos de Estrés por Calor , Conejos , Animales , Antioxidantes/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Proteína 1 Asociada A ECH Tipo Kelch , Estrés Oxidativo , Respuesta al Choque Térmico , Inflamación , Trastornos de Estrés por Calor/veterinaria
2.
Neurochem Res ; 48(10): 3113-3128, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37338792

RESUMEN

More and more evidence shows that the pathological mechanism of vascular dementia (VD) is closely related to oxidative stress injury, cell apoptosis, autophagy, inflammatory response, excitatory amino acid toxicity, synaptic plasticity change, calcium overload, and other processes. Edaravone dexborneol (EDB) is a new type of neuroprotective agent that can improve the neurological damage caused by an ischemic stroke. Previous studies showed that EDB has effects on synergistic antioxidants and induces anti-apoptotic responses. However, it remains unclear whether EDB can affect apoptosis and autophagy by activating the PI3K/Akt/mTOR signaling pathway and its impact on the neuroglial cells. In this study, we established the VD model of rats by bilateral carotid artery occlusion to explore the neuroprotective effect of EDB and its mechanism. Morris Water Maze test was applied to assess the cognitive function of rats. H&E and TUNEL staining were applied to observe the cellular structure of the hippocampus. Immunofluorescence labeling was used to observe the proliferation of astrocytes and microglia. ELISA was applied to examine the levels of TNF-α, IL-1ß and IL-6, and RT-PCR was applied to examine their mRNA expression levels. Western blotting was applied to examine apoptosis-related proteins (Bax, Bcl-2, Caspase-3), autophagy-related proteins (Beclin-1, P62, LC3B), PI3K/Akt/mTOR signaling pathway proteins and their phosphorylation levels. The results indicated that EDB ameliorates learning and memory in rats subjected to the VD model, alleviates neuroinflammatory response by reducing the proliferation of the neuroglial cell and inhibits apoptosis and autophagy, which may be mediated by the PI3K/Akt/mTOR signaling pathway.


Asunto(s)
Demencia Vascular , Fármacos Neuroprotectores , Ratas , Animales , Demencia Vascular/tratamiento farmacológico , Proteínas Proto-Oncogénicas c-akt/metabolismo , Edaravona/farmacología , Ratas Sprague-Dawley , Fosfatidilinositol 3-Quinasas/metabolismo , Enfermedades Neuroinflamatorias , Serina-Treonina Quinasas TOR/metabolismo , Proliferación Celular , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Apoptosis , Autofagia
3.
Zhongguo Zhong Yao Za Zhi ; 47(23): 6380-6390, 2022 Dec.
Artículo en Zh | MEDLINE | ID: mdl-36604883

RESUMEN

Wuling Capsules is one of the commonly used drugs for the clinical treatment of chronic hepatitis B with the syndrome of liver Qi stagnation, spleen deficiency, and blood stasis. However, the present preparation method of Wuling Capsules ignores some macromolecules like polysaccharides. In this study, the influences of different ethanol concentrations in the preparation process on the extraction rates of macro-and micro-molecules were investigated. Further, the therapeutic efficacy of Wuling Capsules was evaluated with the reserpine-induced rat model of liver Qi stagnation, spleen deficiency, and blood stasis. When 50% ethanol was used for the last time of extraction, the concentrations of polysaccharides, salvianolic acid B, and schisandrin in the extract, as well as the dry extract yield, increased significantly compared with those of the original preparation method. However, the fingerprints of micro-molecules showed little difference between the two methods, with a similarity of 0.862. The study then set the 50% ethanol extraction as the new preparation method. The pharmacodynamics evaluation showed that the Wuling Capsules prepared with the original and new methods both significantly alleviated the emotional depression and metabolic disturbance in model rats, demonstrating good performance in protecting the rats against gastric mucosal injuries, modulating intestinal function, and activating blood circulation. The mechanism of action may be related to the regulation of gastrointestinal hormone secretion, reduction of inflammation, and promotion of dopamine synthesis in cortex and hippocampus. At the same dose, the Wuling Capsules prepared with the original and new methods showed roughly the same overall therapeutic efficacy. However, the Wuling Capsules prepared with the new method had stronger effect in activating blood circulation and modulating inflammation, but weaker effects in regulating gastrin and dopamine. The present study provides basis data for optimizing the preparation process of Wuling Capsules and deciphering the mechanism of its therapeutic effect on liver Qi stagnation, spleen deficiency, and blood stasis.


Asunto(s)
Medicina Tradicional China , Qi , Animales , Ratas , Bazo , Cápsulas , Dopamina , Síndrome , Hígado , Inflamación
4.
J Dairy Sci ; 104(3): 2855-2865, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33455801

RESUMEN

Aroma is an important property of fermented milk, and it directly affects consumer acceptance. However, previous studies have mainly focused on analyzing the composition of aroma compounds in fermented milk in vitro, and the composition may be different from the real aroma composition that stimulates the sense of smell. Furthermore, the relationship between olfactory attributes and the release of aroma compounds was not fully understood. In this study, we selected 6 samples of fermented milk differing in aroma perception intensity based on our pretest. A descriptive sensory analysis focusing on orthonasal and retronasal olfaction of fermented milk was first conducted by semitrained panelists. Artificial saliva was mixed with the fermented milk samples and continuously stirred at 37°C for 15 s to simulate oral processing conditions. Headspace solid-phase microextraction-gas chromatography coupled with quadrupole time-of-flight mass spectrometry was applied to identify the head space composition of 6 kinds of fermented milk before and after the simulated oral processing. Twenty-five volatile compounds were identified in the fermented milks, 15 of which were predicted to have an influence on the olfactory perception of fermented milks during oral processing. Partial least squares regression analysis based on chemical and sensory data was then applied to explore the correlation between sensory perception and volatile aroma release. The results showed that oral processing greatly increased the perception of creamy aroma compounds, such as diacetyl and acetone, but did not increase the perception of dairy sour aroma compounds, such as butanoic acid and hexanoic acid. This study can help improve our understanding of the relationship between olfactory perceptions and the release of volatile aroma compounds under oral processing. It might also contribute to the design of palatable fermented milks catering to specific consumer preferences.


Asunto(s)
Percepción Olfatoria , Compuestos Orgánicos Volátiles , Animales , Cromatografía de Gases y Espectrometría de Masas/veterinaria , Odorantes/análisis , Olfato , Gusto , Compuestos Orgánicos Volátiles/análisis
5.
Arch Environ Contam Toxicol ; 81(4): 553-563, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-33386941

RESUMEN

The concentrations of dissolved yttrium and rare earth elements (REY) in sediment pore water provide important geochemical information. However, due to the low REY concentration, complex matrix, and limited sample volume (often only a few milliliters), analysis of the REY in pore water often is highly challenging. In this study, a method was established to determine the dissolved REY in pore water of marine sediments using an offline preconcentration step with the ethylenediaminetriacetate chelating resin, followed by inductively coupled plasma-mass spectrometry. In addition, using a commercially available automated trace-element preconcentration system, the preconcentration step can be fully automated, saving labor and providing a better control of the final elution volume. The experimental conditions (pH, elution volume, elution acid concentration, and organic complexation effect) were assessed, and the optimal conditions were chosen. In particular, the organic complexation effect was found to be negligible. The procedure blank and limit of detection were satisfactory for studying REY in pore water of marine sediments, and the method also yielded satisfactory recoveries for the REY elements (83-110%). The method was then applied to analyze the dissolved REY concentrations of pore water samples collected in a sediment core (~ 30 cm) in the central Indian Ocean. The vertical distribution, dissolved REY concentration, and the average Post Archean Australian Shale-normalized pattern of the REY showed similarities to the previously published pore water REY data. This method provides an accurate yet facile approach for the analysis of all 15 REY in marine pore water samples using the sample volume of only ~ 5 mL.


Asunto(s)
Metales de Tierras Raras , Oligoelementos , Australia , Sedimentos Geológicos , Oligoelementos/análisis , Agua
6.
J Nanobiotechnology ; 18(1): 62, 2020 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-32316985

RESUMEN

Nucleic acid is the main material for storing, copying, and transmitting genetic information. Gene sequencing is of great significance in DNA damage research, gene therapy, mutation analysis, bacterial infection, drug development, and clinical diagnosis. Gene detection has a wide range of applications, such as environmental, biomedical, pharmaceutical, agriculture and forensic medicine to name a few. Compared with Sanger sequencing, high-throughput sequencing technology has the advantages of larger output, high resolution, and low cost which greatly promotes the application of sequencing technology in life science research. Magnetic nanoparticles, as an important part of nanomaterials, have been widely used in various applications because of their good dispersion, high surface area, low cost, easy separation in buffer systems and signal detection. Based on the above, the application of magnetic nanoparticles in nucleic acid detection was reviewed.


Asunto(s)
Nanopartículas de Magnetita/química , Ácidos Nucleicos/análisis , Bacterias/genética , ADN Bacteriano/análisis , ADN Bacteriano/aislamiento & purificación , ADN Bacteriano/metabolismo , Óxido Ferrosoférrico/química , Humanos , MicroARNs/análisis , Técnicas de Amplificación de Ácido Nucleico , Ácidos Nucleicos/aislamiento & purificación , Polimorfismo de Nucleótido Simple
7.
Zhongguo Zhong Yao Za Zhi ; 45(9): 2115-2121, 2020 May.
Artículo en Zh | MEDLINE | ID: mdl-32495560

RESUMEN

A rapid analysis method based on ultraviolet-visual(UV-Vis) spectroscopy, near infrared(NIR) spectroscopy and multivariable data analysis was established for quality evaluation of Shengxuebao Mixture. The contents of eight active ingredients of Shengxuebao Mixture including albiflorin, paeoniflorin, 2, 3, 5, 4'-tetra-hydroxy-stilbene-2-O-ß-D-glucopyranoside, specnuezhenide,ecliptasaponin D, emodin, calycosin-7-glucoside and astragaloside Ⅳ were simultaneously detected by using this method. HPLC-UV-MS was used as a reference method for determining the contents of these ingredients. Partial least squares(PLS) analysis was implemented as a linear method for multivariate models calibrated between UV spectrum/NIR spectrum and contents of 8 ingredients. Finally, the performance of the model was evaluated by 24 batches of test samples. The results showed that both UV-Vis and NIR models gave a good calibration ability with an R~2 value above 0.9, and the prediction ability was also satisfactory, with an R~2 value higher than 0.83 for UV-Vis model and higher than 0.79 for NIR model. The overall results demonstrate that the established method is accurate, robust and fast, therefore, it can be used for rapid quality evaluation of Shengxuebao Mixture.


Asunto(s)
Medicamentos Herbarios Chinos , Espectroscopía Infrarroja Corta , Calibración , Cromatografía Líquida de Alta Presión , Análisis de los Mínimos Cuadrados , Espectrometría de Masas
8.
J Transl Med ; 17(1): 47, 2019 02 18.
Artículo en Inglés | MEDLINE | ID: mdl-30777078

RESUMEN

BACKGROUND: Immunology research, particularly next generation sequencing (NGS) of the immune T-cell receptor ß (TCRß) repertoire, has advanced progression in several fields, including treatment of various cancers and autoimmune diseases. This study aimed to identify the TCR repertoires from dry blood spots (DBS), a method that will help collecting real-world data for biomarker applications. METHODS: Finger-prick blood was collected onto a Whatman filter card. RNA was extracted from DBS of the filter card, and fully automated multiplex PCR was performed to generate a TCRß chain library for next generation sequencing (NGS) analysis of unique CDR3s (uCDR3). RESULTS: We demonstrated that the dominant clonotypes from the DBS results recapitulated those found in whole blood. According to the statistical analysis and laboratory confirmation, 40 of 2-mm punch disks from the filter cards were enough to detect the shared top clones and have strong correlation in the uCDR3 discovery with whole blood. uCDR3 discovery was neither affected by storage temperatures (room temperature versus - 20 °C) nor storage durations (1, 14, and 28 days) when compared to whole blood. About 74-90% of top 50 uCDR3 clones of whole blood could also be detected from DBS. A low rate of clonotype sharing, 0.03-1.5%, was found among different individuals. CONCLUSIONS: The DBS-based TCR repertoire profiling method is minimally invasive, provides convenient sampling, and incorporates fully automated library preparation. The system is sensitive to low RNA input, and the results are highly correlated with whole blood uCDR3 discovery allowing study scale-up to better understand the relationship and mutual influences between the immune and diseases.


Asunto(s)
Pruebas con Sangre Seca/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Regiones Determinantes de Complementariedad/genética , Humanos , Preservación Biológica , ARN/aislamiento & purificación , Temperatura
9.
J Nanosci Nanotechnol ; 19(4): 2054-2059, 2019 04 01.
Artículo en Inglés | MEDLINE | ID: mdl-30486947

RESUMEN

To develop a software program used for controlling the portable on-site pathogen detection system which is based on real-time PCR, this software is developed by C# over the Microsoft Visual Studio 2013 and used on local computers which are equipped with Windows systems. Taking the actual demand as the guidance, it constructs according to the framework design and unifies the barcode technology, the cloud service, and the Bluetooth technology. Based on the above methods, this software can meet the demands of the system such as free experiment design, auto-control of the hardware and efficient data management. During the operation of this program, the serial port and network communication between it and the device and web server remains stable, no matter it is processing the data collection or data analysis and display. It is feasible and practical to apply this software to the portable on-site pathogen detection system in clinical practices and research work.


Asunto(s)
Computadores , Programas Informáticos
10.
J Nanosci Nanotechnol ; 19(4): 2049-2053, 2019 04 01.
Artículo en Inglés | MEDLINE | ID: mdl-30486946

RESUMEN

Micro-liquid distribution operation has been a core part of the fully automatic liquid handling platform in clinical testing. Accurate and fast automatic detection of liquid level with disposable tip is the precondition to ensure the stability of the whole operation of liquid handling platform. By further research on the liquid level sensing technology of pressure sensor, in this paper we designed a new type of accurate and stable micro-liquid level detection system and verified its stability, instantaneity and accuracy by setting up a small reagent distribution platform. It was shown that the liquid level detection system designed in this paper has reliable stability, good instantaneity and high accuracy, with the liquid level detecting accuracy up to 100%, the response speed after detecting the liquid level reaching millisecond and the depth in the liquid level less than 0.3 mm. Besides, there is no limit on the physical and chemical properties of the liquids to be tested, which is consistent with the technical indicators of the liquid handling platform.

11.
Zhongguo Zhong Yao Za Zhi ; 43(21): 4248-4254, 2018 Nov.
Artículo en Zh | MEDLINE | ID: mdl-30583625

RESUMEN

In this study, the HPLC-UV-MS method for the simultaneous determination of eight active ingredients of Shengxuebao Mixture were developed based on the concept of quality by design(QbD)with a stepwise optimization approach. After the analytical target profile(ATP)had been defined, albiflorin, paeoniflorin, 2, 3, 5, 4'-tetra-hydroxy-stilbene-2-O-ß-D-glucopyranoside, specnuezhenide, ecliptasaponin D, emodin, calycosin-7-glucoside, and astragaloside Ⅳ were identified as the indicator components. The resolution and the signal-to-noise ratio of indicator components were then selected as critical method attributes (CMA) for the first step optimization. According to the results collected from fractional factorial design, critical method parameters (CMP) were determined with a multiple linear regression method, which included the amount of acid addition in the mobile phase, temperature, gradient, and wavelength. After that, the amount of acid addition and the wavelength were optimized to improve the resolution and the signal-to-noise ratio of the indicator components. The peak symmetry factors of specnuezhenide and emodin were then set as CMA for the second step optimization. The Box-Behnken designed experiments were conducted. The temperature and gradient were optimized after modelling. The design space were calculated and verified. The optimized analytical method was validated, and the results showed a good precision, accuracy and stability, which means that it can be used for the quantification of the indicator components in Shengxuebao Mixture.


Asunto(s)
Medicamentos Herbarios Chinos/análisis , Fitoquímicos/análisis , Cromatografía Líquida de Alta Presión , Espectrometría de Masas , Reproducibilidad de los Resultados
12.
BMC Microbiol ; 15: 54, 2015 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-25887661

RESUMEN

BACKGROUND: Bifidobacteria are key commensals in human gut, and their abundance is associated with the health of their hosts. Although they are dominant in infant gut, their number becomes lower in adult gut. The changes of the diet are considered to be main reason for this difference. Large amounts of whole-genomic sequence data of bifidobacteria make it possible to elucidate the genetic interpretation of their adaptation to the nutrient environment. Among the nutrients in human gut, starch is a highly fermentable substrate and can exert beneficial effects by increasing bifidobacteria and/or being fermented to short chain fatty acids. RESULTS: In order to determine the potential substrate preference of bifidobacteria, we compared the glycoside hydrolase (GH) profiles of a pooled-bifidobacterial genome (PBG) with a representative microbiome (RM) of the human gut. In bifidobacterial genomes, only 15% of GHs contained signal peptides, suggesting their weakness in utilization of complex carbohydrate, such as plant cell wall polysaccharides. However, compared with other intestinal bacteria, bifidobacteiral genomes encoded more GH genes for degrading starch and starch hydrolysates, indicating that they have genetic advantages in utilizing these substrates. Bifidobacterium longum subsp. longum BBMN68 isolated from centenarian's faeces was used as a model strain to further investigate the carbohydrate utilization. The pathway for degrading starch and starch hydrolysates was the only complete pathway for complex carbohydrates in human gut. It is noteworthy that all of the GH genes for degrading starch and starch hydrolysates in the BBMN68 genome were conserved in all studied bifidobacterial strains. The in silico analyses of BBMN68 were further confirmed by growth experiments, proteomic and real-time quantitative PCR (RT-PCR) analyses. CONCLUSIONS: Our results demonstrated that starch and starch hydrolysates were the most universal and favorable carbon sources for bifidobacteria. The low amount of these carbon sources in adult intestine was speculated to contribute to the low relative abundance of bifidobacteria.


Asunto(s)
Bifidobacterium/metabolismo , Carbono/metabolismo , Tracto Gastrointestinal/microbiología , Almidón/metabolismo , Bifidobacterium/genética , Bifidobacterium/aislamiento & purificación , Metabolismo de los Hidratos de Carbono , Humanos , Hidrólisis , Redes y Vías Metabólicas/genética
13.
J Nanosci Nanotechnol ; 15(7): 4815-28, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-26373042

RESUMEN

China is one of the countries with the highest incidence of gastric cancer, and accounts for over 40% of all new gastric cancer cases in the world. Genetic factors as well as environmental factors play a role in development of gastric cancer. To investigate the independent roles of single nucleotide polymorphisms (SNPs) in base excision repair (BER) genes (APE1 and NEIL2), carcinogen metabolism gene (CYP2E1) and tumor suppressor pathway gene (MDM2) for gastric cancer susceptibility in a Chinese population, we conducted a hospital based case-control study to evaluate the potential association between these polymorphisms and susceptibility to gastric cancer in a Northern Jiangsu population. We also associated the NEIL-2 mRNA expression with the studied NEIL2 SNP genotypes to assess whether the genotypes have influence on the NEIL2 mRNA (hence protein) expression. Five SNPs, APE 1 (rs2275008), NEIL 2 (rs804270), MDM2 (rs2279744), and CYP 2E1 (rs2480256 and rs2031920), were genotyped by TaqMan assays in 105 gastric cancer cases and 118 controls. Genotype frequency distribution showed that the APE 1 SNP (rs2275008), NEIL 2 SNP (rs804270), MDM2 SNP (rs2279744), and CYP 2E1 SNP (rs2031920) had more mutant alleles in gastric cancer cases than controls (76.19, 68.57, 54.29, and 43.81%, respectively), while CYP 2E1 SNP (rs2480256) had large percentage of both alleles (43.81%). Risk analysis revealed that there was increased risk for gastric cancer in subjects with mutant alleles in APE 1 (rs2275008: OR 5.49, 95% CI = 2.6-5.7, p <.0001), NEIL 2 (rs804270: OR 2.3, 95% CI = 1.22-4.3, p=0.01), MDM2 (rs2279744: OR 14.65, 95% CI = 5.63-8.15, p < .0001), and CYP 2E1 (rs2031920: OR 8.385, 95% CI = 3.2-5.3, p < .0001) SNPs. Moreover, the NEIL2 mRNA expression analysis showed that there was significant differential expression of NEIL2 mRNA among the randomly tested NEIL2 genotypes (p = 0.005), with low expression seen in variant genotypes than in other genotypes. In conclusion, variant alleles in the NEIL2 (rs804270), APE1 (rs2275008), CYP2E1 (rs2031920) and MDM2 (rs2279744) SNPs may independently influence susceptibility to gastric cancer in a Northern Jiangsu Chinese population. The genotypes may also independently influence their respective gene mRNA expression, as seen in our study, where there was differential expression of the NEIL2 mRNA among the genotypes, with low NEIL2 mRNA expression seen in the variant genotype.


Asunto(s)
Citocromo P-450 CYP2E1/genética , ADN Glicosilasas/genética , ADN-(Sitio Apurínico o Apirimidínico) Liasa/genética , Predisposición Genética a la Enfermedad , Polimorfismo de Nucleótido Simple , Proteínas Proto-Oncogénicas c-mdm2/genética , Neoplasias Gástricas/genética , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Pueblo Asiatico , Estudios de Casos y Controles , China/epidemiología , Citocromo P-450 CYP2E1/biosíntesis , ADN Glicosilasas/biosíntesis , ADN-(Sitio Apurínico o Apirimidínico) Liasa/biosíntesis , Femenino , Regulación de la Expresión Génica , Frecuencia de los Genes , Humanos , Masculino , Persona de Mediana Edad , Proteínas Proto-Oncogénicas c-mdm2/biosíntesis , Neoplasias Gástricas/epidemiología , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología
14.
J Nanosci Nanotechnol ; 15(2): 1123-9, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26353621

RESUMEN

In this research, a novel method for relative fluorescent quantification of DNA based on Fe3O4@SiO2@Au gold-coated magnetic nanocomposites (GMNPs) and multiplex ligation- dependent probe amplification (MLPA) has been developed. With the help of self-assembly, seed-mediated growth and chemical reduction method, core-shell Fe3O4@SiO2@Au GMNPs were synthesized. Through modified streptavidin on the GMNPs surface, we obtained a bead chip which can capture the biotinylated probes. Then we designed MLPA probes which were tagged with biotin or Cy3 and target DNA on the basis of human APP gene sequence. The products from the thermostable DNA ligase induced ligation reactions and PCR amplifications were incubated with SA-GMNPs. After washing, magnetic separation, spotting, the fluorescent scanning results showed our method can be used for the relative quantitative analysis of the target DNA in the concentration range of 03004~0.5 µM.


Asunto(s)
ADN/análisis , ADN/genética , Nanopartículas del Metal/química , Microscopía Fluorescente/instrumentación , Reacción en Cadena de la Polimerasa Multiplex/instrumentación , Nanocompuestos/ultraestructura , Secuencia de Bases , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Diseño de Equipo , Análisis de Falla de Equipo , Oro/química , Nanopartículas del Metal/ultraestructura , Microscopía Fluorescente/métodos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa Multiplex/métodos , Nanocompuestos/química , Análisis de Secuencia de ADN/instrumentación , Análisis de Secuencia de ADN/métodos , Dióxido de Silicio/química
15.
J Nanosci Nanotechnol ; 14(5): 3337-42, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24734549

RESUMEN

A rapid, ultrasensitive and economical Pseudorabies virus (PRV) detection system based on magnetic beads (MBs) and chemiluminescence was developed in this paper. The carboxyl functionalized MBs (MBs-COOH) were covalently coupled with aminated DNA probes for capturing PRV biotinylated amplicon, the product of polymerase chain reaction (PCR). Agarose gel electrophoresis analysis approved the reliability of biotinylated amplicon. The MBs composites were incubated with alkaline phosphatase labeled streptavidin (ALP-SA) and chemiluminescene was determined by subsequently adding 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl-1,2-dioxetane (AMPPD). The optimal conditions of the PRV detection method were 10 microM for probe concentration, 50 degrees C for hybridization temperature and 30 min for hybridization time. The limit of detection (LOD) was as low as 100 amol/5 pM of amplicon which proved that this approach for PRV detection was ultrasensitive.


Asunto(s)
Herpesvirus Suido 1/aislamiento & purificación , Magnetismo , Secuencia de Bases , Cartilla de ADN , Sondas de ADN , Electroforesis en Gel de Agar , Herpesvirus Suido 1/genética , Humanos , Límite de Detección , Luminiscencia , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa
16.
Guang Pu Xue Yu Guang Pu Fen Xi ; 34(1): 92-7, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24783540

RESUMEN

The effects of calcium ion on the structural properties of casein micelles in the course of heat treatment were synthetically examined by non-structure-invasive spectrometry. The hydrophobicity, reflected by extrinsic fluorescence (ANS fluorescence), was positively correlated with the concentration of the calcium ion, within the range of 0 to 12 mmol x L(-1). Meanwhile, the turbidity and stability of casein micelles also increased with the growth of calcium concentrations. However, opposite results were observed for hydrodynamic diameter and polydispersity index. Compared with the calcium ion, the calcium-chelator (citrate) has an opposite effect on the structural characteristics of casein micelles. Within the calcium concentrations range of 0 to 12 mmol x L(-1), the hydrophobicity, stability and turbidity were negatively correlated with the concentration of the calcium ion, nevertheless, opposite results were observed for hydrodynamic diameter and polydispersity index. All the results indicate that the calcium ion could be used to modify the structures of casein micelles during heat heatment.


Asunto(s)
Calcio/química , Caseínas/química , Micelas , Ácido Cítrico , Calor , Interacciones Hidrofóbicas e Hidrofílicas , Iones , Análisis Espectral
17.
Front Bioeng Biotechnol ; 12: 1393789, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38725992

RESUMEN

The significance of point-of-care testing (POCT) in early clinical diagnosis and personalized patient care is increasingly recognized as a crucial tool in reducing disease outbreaks and improving patient survival rates. Within the realm of POCT, biosensors utilizing magnetic nanoparticles (MNPs) have emerged as a subject of substantial interest. This review aims to provide a comprehensive evaluation of the current landscape of POCT, emphasizing its growing significance within clinical practice. Subsequently, the current status of the combination of MNPs in the Biological detection has been presented. Furthermore, it delves into the specific domain of MNP-based biosensors, assessing their potential impact on POCT. By combining existing research and spotlighting pivotal discoveries, this review enhances our comprehension of the advancements and promising prospects offered by MNP-based biosensors in the context of POCT. It seeks to facilitate informed decision-making among healthcare professionals and researchers while also promoting further exploration in this promising field of study.

18.
Indian J Microbiol ; 53(4): 492-5, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24426157

RESUMEN

In order to improve the production of the milk-clotting enzyme under submerged fermentation, two statistical methods were applied to optimize the culture conditions of Bacillus amyloliquefaciens D4 using wheat bran as nutrient source. First, initial pH, agitation speed, and fermentation time were shown to have significant effects on D4 enzyme production using the Plackett-Burman experimental design. Subsequently, optimal conditions were obtained using the Box-Behnken method, which were as follows: initial pH 7.57, agitation speed 241 rpm, fermentation time 53.3 h. Under these conditions, the milk-clotting enzyme production was remarkably enhanced. The milk-clotting enzyme activity reached 1996.9 SU/mL, which was 2.92-fold higher than that of the initial culture conditions, showing that the Plackett-Burman design and Box-Behnken response surface method are effective to optimize culture conditions. The research can provide a reference for full utilization of wheat bran and the production of milk-clotting enzyme by B. amyloliquefaciens D4 under submerged fermentation.

19.
Transl Neurosci ; 14(1): 20220295, 2023 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-37600117

RESUMEN

Objective: This research was conducted to discuss the recent prognosis of patients with acute cerebral infarction (ACI) combined with cerebral-cardiac syndrome (CCS). Method: Eighty-seven patients with ACI were selected, which were divided into the ACI group (52 patients) and the CCS group (35 patients) according to whether the CCS was combined, and another 30 health controls were selected as the control group. Serum hypoxia-inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF) levels of subjects in each group at the 1st day, the 3rd day, and the 7th day after admission were measured by enzyme-linked immunosorbent assay. After discharge for 30 days, the National Institutes of Health Stroke Scale (NIHSS) and the modified Rankin Scale (mRS) score were utilized to evaluate the prognosis of patients. The role of serum HIF-1α and VEGF levels in the prognosis of ACI combined with CCS patients was assessed by receiver operating characteristic curve and the binary logistic regression analysis. Results: Higher serum HIF-1α and VEGF levels were observed in the CCS and ACI groups versus the control group, and the levels of which were even higher in the CCS group in comparison to the ACI group. According to the prognosis of the NIHSS score, fasting blood glucose (FBG), Acute Physiology and Chronic Health Evaluation II score, creatine kinase-MB (CK-MB), and HIF-1α and VEGF levels at the 7th day of admission were higher while Glasgow coma scale (GCS) score was lower in the poor prognosis group than those in the good prognosis group, and the area under the curve (AUC) of serum HIF-1α and VEGF levels was 0.895 (95% confident interval [CI], 0.786-1.000), and 0.855 (95% CI, 0.731-0.980). According to the prognosis of the mRS score, FBG, CK-MB, and HIF-1α and VEGF levels at the 7th day of admission were higher while GCS score was lower in the poor prognosis group than those in the good prognosis group, and the AUC of serum HIF-1α and VEGF levels was 0.850 (95% CI, 0.722-0.979) and 0.901 (95% CI, 0.798-1.000). The results of the binary logistic regression analysis revealed that HIF-1α and VEGF levels may be independent risk factors influencing the prognosis of ACI combined with CCS. Conclusion: Serum HIF-1α and VEGF have a good predictive value for assessing the recent prognosis of patients with ACI combined with CCS, which could be independent risk factors influencing the prognosis of disease.

20.
Biosensors (Basel) ; 13(2)2023 Jan 19.
Artículo en Inglés | MEDLINE | ID: mdl-36831926

RESUMEN

Infectious diseases and tumors have become the biggest medical challenges in the 21st century. They are driven by multiple factors such as population growth, aging, climate change, genetic predispositions and more. Nucleic acid amplification technologies (NAATs) are used for rapid and accurate diagnostic testing, providing critical information in order to facilitate better follow-up treatment and prognosis. NAATs are widely used due their high sensitivity, specificity, rapid amplification and detection. It should be noted that different NAATs can be selected according to different environments and research fields; for example, isothermal amplification with a simple operation can be preferred in developing countries or resource-poor areas. In the field of translational medicine, CRISPR has shown great prospects. The core component of NAAT lies in the activity of different enzymes. As the most critical material of nucleic acid amplification, the key role of the enzyme is self-evident, playing the upmost important role in molecular diagnosis. In this review, several common enzymes used in NAATs are compared and described in detail. Furthermore, we summarize both the advances and common issues of NAATs in clinical application.


Asunto(s)
Enfermedades Transmisibles , Ácidos Nucleicos , Humanos , Técnicas de Amplificación de Ácido Nucleico
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