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In this study, we applied various thermal pretreatment methods (e.g., hot-air, microwave, and stir-frying) to process walnut kernels, and conducted comparative analysis of the physicochemical properties, nutritional components, in vitro antioxidant activity, and flavor substances of the extracted walnut oil (WO). The results indicated that, thermal pretreatment significantly increased the extraction of total trace nutrients (e.g., total phenols, tocopherols, and phytosterols) in WO. The WO produced using microwave had 2316.71 mg/kg of total trace nutrients, closely followed by the stir-frying method, which yielded an 11.22% increase compared to the untreated method. The WO obtained by the microwave method had a higher Oxidative inductance period (4.05 h) and oil yield (2.48%). After analyzing the flavor in WO, we found that aldehydes accounted for 28.77% of the 73 of volatile compounds and 58.12% of the total flavor compound content in microwave-pretreated WO, these percentages were higher than those recorded by using other methods. Based on the comprehensive score obtained by the PCA, microwave-pretreatment might be a promising strategy to improve the quality of WO based on aromatic characteristics.
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Aromatizantes , Juglans , Oxirredução , Óleos de Plantas , Paladar , Compostos Orgânicos Voláteis , Juglans/química , Compostos Orgânicos Voláteis/química , Compostos Orgânicos Voláteis/análise , Aromatizantes/química , Aromatizantes/análise , Óleos de Plantas/química , Antioxidantes/análise , Antioxidantes/química , Temperatura Alta , Micro-OndasRESUMO
The health benefits of fermented fruits have attracted consumers' attention. High levels of antioxidant ability in the fermented kiwifruit extract were found at the early stage of fermentation. The co-fermention with Lactobacillus paracasei LG0260 and Kluyveromyces marxianus J2853 showed the highest ABTS radical scavenging ability (ABTSâ +-SA) and superoxide dismutase (SOD) activity. Also, the typical antioxidant components of SOD activity, vitamin C concentration and total phenol content were highly correlated with ABTSâ +-SA. Obviously, polyphenols in the fermented kiwifruit extract evolved into monophenols during fermentation. Compared to undigested samples, the activity of ABTSâ +-SA and reducing power capacity (RP-CA) after the final intestinal digestion decreased and ranged 387.44-531.89 VCµg/mL, 650.95-981.63 VCµg/mL, respectively (P < 0.05). Meanwhile, SOD activity on the 10th day of fermentation were still remained 222.82 U/mL, 206.98 U/mL and 217.23 U/mL, respectively. These results suggested that the fermented kiwifruit extract could exhibit antioxidant activity through tolerance to the digestive environment.
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Antioxidantes , Benzotiazóis , Lactobacillales , Ácidos Sulfônicos , Antioxidantes/metabolismo , Lactobacillales/metabolismo , Extratos Vegetais , Superóxido Dismutase , Digestão , FermentaçãoRESUMO
Walnut oils were obtained by supercritical carbon dioxide extraction (SCB), cold-pressing (CP), hexane extraction (HE), and subcritical butane extraction (SBE), and walnut protein isolates (WPI) from the walnut cakes were performed. The results indicate that SCB has the highest oil yield for walnut oil, which was 62.72%, and the total content of trace nutrients (total tocopherols, total phytosterols, and total phenolic compounds) in SCB-walnut oil was also the highest at 2186.75 mg/kg, approximately 1.05 times higher than CP-walnut oil and 1.21 times higher than SBE-walnut oil. Meanwhile, the treatment of WPI with SCB results in a decrease in ß-Sheet and α-Helix structures and an increase in ß-Turn and Random coil structures. Thereby increasing its oil-holding capacity (OHC) and solubility by approximately 1.16 times and 1.27 times compared to CP, respectively. Interestingly, SCB as a green oil production technology, also has good prospects for retaining WPI functionality characteristics.
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Juglans , Juglans/química , Óleos de Plantas/química , Tocoferóis , Antioxidantes/química , NutrientesRESUMO
Phosphogypsum (PG), which has great environmental harm and restricts the sustainable development of phosphorus chemical industry, is a solid waste produced in phosphoric acid production. Thermal decomposition of PG is an extensive way to reutilization of resource, and the key point is to establish an appropriate decomposition path and catalyst system of PG. In the work, the strategy for low-temperature and highly-efficient decomposition of PG is established based on the thermodynamic analysis and the experimental research by metal ions to reduce the decomposition temperature. Meanwhile, SEM(Scanning Electron Microscope) is used to characterize the composition and morphology of PG in the various conditions, also the decomposition temperature is analyzed by TGA(Thermogravimetric Analysis). Then, the decomposition ratio via Fe3+/Co2+/Ni2+ unitary/ternary catalyst is obtained by precipitation method. Through kinetic analysis combined with XRF(X-ray Fluorescence Spectrometer) and EDX(Energy Dispersive X-Ray Spectroscopy) results, it is found that there is a reaction competition in the decomposition process by Fe3+/Co2+/Ni2+ ternary catalyst. Further the mechanism of catalytic system on PG is derived. The present work can be concluded that Fe3+/Co2+/Ni2+ can effectively reduce the decomposition temperature of PG, and the effect of ternary metal is more obvious than that of unitary metal. Finally, pomelo peel is used instead of coke to successfully decompose PG at low temperature by one step method. The establishment of low temperature decomposition system of PG has potential application in phosphorus chemical industry and is in line with sustainable development.
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Metais , Fósforo , Temperatura , Cinética , Fósforo/análiseRESUMO
OBJECTIVE: The drug-refractory epilepsy (DRE) in children is commonly observed but the underlying mechanisms remain elusive. We examined whether fatty acids (FAs) and lipids are potentially associated with the pharmacoresistance to valproic acid (VPA) therapy. METHODS: This single-center, retrospective cohort study was conducted using data from pediatric patients collected between May 2019 and December 2019 at the Children's Hospital of Nanjing Medical University. Ninety plasma samples from 53 responders with VPA monotherapy (RE group) and 37 non-responders with VPA polytherapy (NR group) were collected. Non-targeted metabolomics and lipidomics analysis for those plasma samples were performed to compare the potential differences of small metabolites and lipids between the two groups. Plasma metabolites and lipids passing the threshold of variable importance in projection value >1, fold change >1.2 or <0.8, and p-value <0.05 were regarded as statistically different substances. RESULTS: A total of 204 small metabolites and 433 lipids comprising 16 different lipid subclasses were identified. The well-established partial least squares-discriminant analysis (PLS-DA) revealed a good separation of the RE from the NR group. The FAs and glycerophospholipids status were significantly decreased in the NR group, but their triglycerides (TG) levels were significantly increased. The trend of TG levels in routine laboratory tests was in line with the lipidomics analysis. Meanwhile, cases from the NR group were characterized by a decreased level of citric acid and L-thyroxine, but with an increased level of glucose and 2-oxoglutarate. The top two enriched metabolic pathways involved in the DRE condition were biosynthesis of unsaturated FAs and linoleic acid metabolism. SIGNIFICANCE: The results of this study suggested an association between metabolism of FAs and the medically intractable epilepsy. Such novel findings might propose a potential mechanism linked to the energy metabolism. Ketogenic acid and FAs supplementation might therefore be high-priority strategies for DRE management.
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Epilepsia Resistente a Medicamentos , Humanos , Criança , Triglicerídeos , Epilepsia Resistente a Medicamentos/tratamento farmacológico , Lipidômica , Ácidos Graxos , Estudos Retrospectivos , Ácido Valproico/uso terapêuticoRESUMO
Tumor protein D52-like 2 or simply TPD52L2 belongs to the TPD52 family which has been implicated in a variety of human carcinomas. However, the TPD52L2 function in the gastric carcinoma oxaliplatin (OXA) resistance remains elusive. The main objective of this study is to evaluate the TPD52L2 effect in OXA-resistant gastric carcinoma cells in vitro. Oxaliplatin-resistant gastric carcinoma cells were generated in MGC-803 and SGC-7901 cells. siRNA-mediated knockdown of TPD52L2 was investigated in OXA-resistant MGC-803-OXA and SGC-7901-OXA cells. qRT-PCR was performed to assess the expression level of TPD52L2 mRNA. TPD52L2 protein expression level, apoptosis, and endoplasmic reticulum (ER) stress-associated proteins were identified via immunoblotting analysis. MTT assay was conducted for the evaluation of cell viability, while colony-forming activity was carried out via crystal violet staining. SGC-7901-OXA and MGC-803-OXA cells were found to be more resistant to OXA, as compared to the parental cell lines. The expression of TPD52L2 was found to be upregulated in OXA-resistant cells. Knockdown of TPD52L2 suppressed cell colony-forming potency, cell growth, and development in OXA-resistant cells. TPD52L2 knockdown also enhanced the PARP and caspase-3 cleavage. ER-associated proteins such as PERK, GRP78, CHOP, and IRE1α were found to be elevated in TPD52L2 knockdown cells. ER stress might be involved in TPD52L2 knockdown-induced apoptosis in OXA-resistant gastric carcinoma cells.
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BACKGROUND: Acupuncture is widely used for treating migraine; however, evidence of its efficacy when compared with sham acupuncture is equivocal. This study aimed to compare the evaluate efficacy of acupuncture versus sham acupuncture in migraine treatment. METHODS: We searched PubMed, the Cochrane Library, Web of Science, and EMBASE databases from their inception to April 25, 2021, for randomized controlled trials. The outcome measurement included response rate, migraine days, intensity, and frequency. RESULTS: Twenty randomized controlled trials involving 2725 patients were included. Pooled data suggested that acupuncture was superior to sham acupuncture [mean difference (MD)=-0.52, 95% confidence interval (CI): -0.71 to -0.34, P<0.00001] in reducing the migraine frequency after treatment and follow-up (MD=-0.51, 95% CI: -0.70 to -0.32, P<0.00001). In reducing Visual Analog Scale, acupuncture was superior to sham acupuncture (MD=-0.72, 95% CI: -1.17 to -0.27, P=0.002) after treatment, and also superior to sham acupuncture (MD=-0.82, 95% CI: -1.31 to -0.33, P=0.001) on the follow-up. As for responder rate, acupuncture's efficacy was better than sham acupuncture (relative risk=1.28, 95% CI: 1.00-1.64, P=0.05). However, the reduction of migraine days in the acupuncture group was not significantly different from that in the sham acupuncture group after treatment (MD=-0.62; 95% CI: -1.31 to 0.08; P=0.08) and on the follow-up (MD=-0.68; 95% CI: -1.52 to 0.17; P=0.12). CONCLUSIONS: Acupuncture appears to be an effective treatment modality for migraine. Compared with sham acupuncture, acupuncture had greater improvements in migraine frequency, Visual Analog Scale, and responder rate, but there was no statistically significant difference for the number of migraine days.
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Terapia por Acupuntura , Transtornos de Enxaqueca , Humanos , Transtornos de Enxaqueca/terapia , Revisões Sistemáticas como Assunto , Resultado do TratamentoRESUMO
To explore the action mechanism of Taohong Siwu Decoction(THSWD) in the treatment of soft tissue injury(STI) based on UPLC-Q-TOF-MS technique, network pharmacology and experimental verification method. UPLC-Q-TOF-MS technique was used to identify the chemical constituents of THSWD. The active ingredients and predicted target proteins of THSWD were screened out through TCMSP database. Cytoscape software was used to construct the active component-target-pathway network, and STRING database was used for protein interaction analysis. GeneCards and CTD databases were used to screen out relevant targets of STI. GO function and KEGG pathway enrichment analysis were performed through DAVID database. The rat model of STI was constructed, and Western blot was used to verify the effect of THSWD on key targets of relevant pathways. The results showed 40 active ingredients in THSWD, and 141 potential targets and 20 targets of STI. Target enrichment analysis of the active components produced 128 KEGG pathways, which were mainly concentrated in amino acid synthesis and metabolism, disease signaling pathways, apoptosis, inflammation and other relevant pathways. Western blot showed that THSWD intervention could significantly decrease PTGS2, CASP3, NFKB1, p-CASP3 and p-NFKB1, while enhancing the expression of TP53 protein in the STI samples of rats. According to the results of UPLC-Q-TOF-MS, network pharmacology and experimental verification, active ingredients in THSWD may play anti-inflammatory and antioxidant effects in NF-κB signaling pathway and apoptotic pathway, thus playing a role in the treatment of STI.
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Medicamentos de Ervas Chinesas , Lesões dos Tecidos Moles , Animais , Apoptose , Ratos , Transdução de SinaisRESUMO
BACKGROUND: To observe the therapeutic effect of conventional decompressive craniectomy with hematoma evacuation and frame-based stereotactic minimally invasive surgery (MIS) for supratentorial intracranial hematoma with herniation. METHODS: One hundred forty-nine patients with hypertensive ICH complicated with tentorial herniation were reviewed and analyzed in the present study. The intracranial hematoma was evacuated by emergency surgery within 6 h after admission. According to the authorized representatives' wishes and consent, 74 of the 149 patients were treated by conventional decompressive craniectomy followed by hematoma removal, defined as the CDC group, and the remaining 75 patients underwent frame-based stereotactic MIS for ICH evacuation, defined as the MIS group. The intervals between the admission to surgery, the duration of surgery, the amount of iatrogenic bleeding, the occurrence of postoperative rebleeding, and the recovery of neurological functions were compared between the two groups. All patients were followed up for 3 months. Secondary epilepsy, survival in a vegetative state, severe pulmonary complications, mortality, and activities of daily living (ADL) classification were also recorded and compared. RESULTS: The interval between admission and surgery, the duration of surgery, and intraoperative blood loss in the MIS group were significantly decreased compared to the CDC group. The mortality rate, the rate of rebleeding, prevalence of vegetative state, and severe pulmonary complications in the MIS group were remarkably decreased compared to the CDC group. In the MIS group, the survivors' (ADL) grade also showed advantages. CONCLUSIONS: In the surgical treatment of hypertensive ICH complicated with tentorial herniation, frame-based stereotactic MIS for ICH showed advantages compared to conventional open surgery.
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A novel gill cell line from pearl gentian grouper (Epinephelus lanceolatusâ×Epinephelus fuscoguttatusâ, PGGG cell line) was established, its application in cadmium (Cd) toxicology was demonstrated in this study. Primary cultures and PGGG subcultures were carried out at 25 °C in Dulbecco's Modified Eagle medium/F12 medium (1:1; pH 7.2) supplemented with 15% fetal bovine serum (FBS). Primary PGGG cells were spindle-shaped, proliferated into a confluent monolayer within two weeks and were continuously subcultured over passage 60. The growth of cells at passages 20, 40, and 60 was examined. Chromosome analysis revealed that the chromosomal number of normal PGGG cells was 48, but the number of cells with the normal chromosomes number decreased during the passaging process. Cadmium is one of the most toxic metals in aquatic systems and has been associated with multiple animal and human health problems. To interpret the cytotoxicity and related mechanisms of cadmium, PGGG cells were used as an in vitro model. After treatment with cadmium at concentrations ranging from 1 µM to 500 µM, PGGG cells demonstrated dose- and time-dependent cytotoxicity, manifested as morphological abnormalities and a viability decline. Further, it was found that the reactive oxygen species (ROS) and malondialdehyde (MDA) levels were elevated following cadmium exposure, and related genes involved in the antioxidant system, including those encoding catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and Kelch-like- ECH-associated protein 1 (Keap1), were regulated differently. In addition, PGGG cells treated with cadmium had the typical features associated with apoptosis, including phosphatidylserine (PS) externalization; upregulated expression of caspase-3, -8, and -9; and apoptotic body formation. In general, the PGGG cell line may serve as a useful tool for studying the toxic mechanisms of cadmium or other toxicants or for toxicity testing and environment monitoring.
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Apoptose/efeitos dos fármacos , Bass , Cádmio/toxicidade , Expressão Gênica/efeitos dos fármacos , Brânquias , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Poluentes Químicos da Água/toxicidade , Animais , Antioxidantes/metabolismo , Catalase/genética , Catalase/metabolismo , Técnicas de Cultura de Células , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Brânquias/citologia , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismoRESUMO
A novel acidogenic phosphorus recovery (APR) process was developed in combination with Fe(III)-based chemical phosphorus removal and a membrane bioreactor (MBR) for enhanced wastewater treatment and effective P recovery. Two different system configurations were evaluated: Fe-dosing MBR (Fe-MBR), with the Fe-dosing into the MBR, and Fe-enhanced primary sedimentation followed by the MBR (FeP-MBR). The results show that both systems performed well for enhanced nutrient (N and P) removals and P recovery, with approximately 50% of the total P recovered from the municipal wastewater in the form of vivianite. Compared to the Fe-MBR system, FeP-MBR achieved more efficient P recovery under low food-waste loading conditions, maintained a higher ratio of biomass in activated sludge and experienced a slower rate of membrane fouling. Important functional bacteria were identified, including Prevotella and Selenomonas, which are active in hydrolysis and acidogenesis of sludge, and Aeromonas and Sulfurospirillum, which are involved in dissimilatory iron reduction.
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Reatores Biológicos , Ferro/química , Fósforo/isolamento & purificação , Esgotos/microbiologia , Águas Residuárias/química , Compostos Ferrosos/metabolismo , Hidrólise , Fosfatos/metabolismo , Fósforo/metabolismo , Eliminação de Resíduos Líquidos/métodosRESUMO
Tea polyphenols have received much attention from the pharmaceutical and food industries owing to their extraordinary antioxidant and antibacterial characteristics. However, tea polyphenols are very unstable in processing and storage, since they are sensitive to the environmental factors like temperature, light and pH. Therefore, the effective application of tea polyphenols requires a protective mechanism to maintain its activity. The utilization of compounded tea polyphenols, instead of raw materials, can potentially help to improve their stability. This review focuses on the summarization of the compounding technologies for tea polyphenols, including physical technologies, chemical-interfacial technologies and nano-scale compounding technologies. Of which, the emerging nano cellulose bio-carrier, as a promising technology, is particularly proposed.
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Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Neoplasias/tratamento farmacológico , Polifenóis/farmacologia , Chá/química , Antibacterianos/química , Antineoplásicos/química , Antioxidantes/química , Bactérias/efeitos dos fármacos , Doenças Cardiovasculares/tratamento farmacológico , Estabilidade de Medicamentos , Doenças Neurodegenerativas/tratamento farmacológico , Polifenóis/químicaRESUMO
A major shortcoming of empirical scoring functions is that they often fail to predict binding affinity properly. Removing false positives of docking results is one of the most challenging works in structure-based virtual screening. Postdocking filters, making use of all kinds of experimental structure and activity information, may help in solving the issue. We describe a new method based on detailed protein-ligand interaction decomposition and machine learning. Protein-ligand empirical interaction components (PLEIC) are used as descriptors for support vector machine learning to develop a classification model (PLEIC-SVM) to discriminate false positives from true positives. Experimentally derived activity information is used for model training. An extensive benchmark study on 36 diverse data sets from the DUD-E database has been performed to evaluate the performance of the new method. The results show that the new method performs much better than standard empirical scoring functions in structure-based virtual screening. The trained PLEIC-SVM model is able to capture important interaction patterns between ligand and protein residues for one specific target, which is helpful in discarding false positives in postdocking filtering.
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Avaliação Pré-Clínica de Medicamentos/métodos , Proteínas/metabolismo , Sítios de Ligação , Receptores ErbB/química , Receptores ErbB/metabolismo , Ligantes , Simulação de Acoplamento Molecular , Ligação Proteica , Conformação Proteica , Proteínas/química , Máquina de Vetores de Suporte , Interface Usuário-ComputadorRESUMO
The anticancer agent 3-bromopyruvate (3-BP) is viewed as a glycolytic inhibitor that preferentially kills glycolytic cancer cells through energy depletion. However, its cytotoxic activity is dependent on cellular drug import through transmembrane monocarboxylate transporter 1 (MCT-1), which restricts its anticancer potential to MCT-1-positive tumor cells. We created and characterized an MCT-1-independent analog of 3-BP, called NEO218. NEO218 was synthesized by covalently conjugating 3-BP to perillyl alcohol (POH), a natural monoterpene. The responses of various tumor cell lines to treatment with either compound were characterized in the presence or absence of supplemental pyruvate or antioxidants N-acetyl-cysteine (NAC) and glutathione (GSH). Drug effects on glyceraldehyde 3-phosphate dehydrogenase (GAPDH) enzyme activity were investigated by mass spectrometric analysis. The development of 3-BP resistance was investigated in MCT-1-positive HCT116 colon carcinoma cells in vitro. Our results show that NEO218: (i) pyruvylated GAPDH on all 4 of its cysteine residues and shut down enzymatic activity; (ii) severely lowered cellular ATP content below life-sustaining levels, and (iii) triggered rapid necrosis. Intriguingly, supplemental antioxidants effectively prevented cytotoxic activity of NEO218 as well as 3-BP, but supplemental pyruvate powerfully protected cells only from 3-BP, not from NEO218. Unlike 3-BP, NEO218 exerted its potent cytotoxic activity irrespective of cellular MCT-1 status. Treatment of HCT116 cells with 3-BP resulted in prompt development of resistance, based on the emergence of MCT-1-negative cells. This was not the case with NEO218, and highly 3-BP-resistant cells remained exquisitely sensitive to NEO218. Thus, our study identifies a mechanism by which tumor cells develop rapid resistance to 3-BP, and presents NEO218 as a superior agent not subject to this cellular defense. Furthermore, our results offer alternative interpretations of previously published models on the role of supplemental antioxidants: Rather than quenching reactive oxygen species (ROS), supplemental NAC or GSH directly interact with 3-BP, thereby neutralizing the drug's cytotoxic potential before it can trigger ROS production. Altogether, our study introduces new aspects of the cytotoxic mechanism of 3-BP, and characterizes NEO218 as an analog able to overcome a key cellular defense mechanism towards this drug.
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Antineoplásicos/farmacologia , Resistencia a Medicamentos Antineoplásicos , Transportadores de Ácidos Monocarboxílicos/metabolismo , Monoterpenos/farmacologia , Neoplasias/tratamento farmacológico , Piruvatos/farmacologia , Simportadores/metabolismo , Trifosfato de Adenosina/metabolismo , Alquilação , Antioxidantes/farmacologia , Relação Dose-Resposta a Droga , Gliceraldeído-3-Fosfato Desidrogenases , Glicólise/efeitos dos fármacos , Células HCT116 , Humanos , Células MCF-7 , Transportadores de Ácidos Monocarboxílicos/genética , Necrose , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patologia , Interferência de RNA , Transdução de Sinais/efeitos dos fármacos , Simportadores/genética , TransfecçãoRESUMO
To explore the effects of stand age on variation patterns of leaf C, N, P stoichiometric characteristics of Machilus pauhoi, two stands, i.e., 9 and 13 years old, were selected. The relationships between leaf nutrient contents (C, N and P) and diameters at breast height (DBH) of individual plants were analyzed. The data revealed that the individual variations of seedlings in M. pauhoi stands were strengthened with the stand development. The stand age had significant effects on leaf C, N, P contents and C:N ratio but not on C:P and N: P ratios. Specifically, the mean values of leaf C, N, P contents and N:P ratio in the 9-year-old stand were lower than those in the 13-year-old stand, whereas, inverse pattern of C:N and C:P ratios were found in the two stands. Furthermore, leaf N and P stoichiometry varied significantly within the stand. Specifically, leaf N and P contents, as well as their stoichiometric ratios, linearly correlated with DBH in the 9-year-old stand. On the contrary, leaf N and P stoichiometry showed quadratic correlation in 13-year-old stand (except leaf C:N which linearly correlated with DBH). Lastly, nutrient transfer rates of leaf N and P in the 9-year-old stand were higher than that in 13-year-old stand, and the discrepancies of leaf nutrient transfer strategy between growing and non-growing seasons were caused by the different growth phases and environmental conditions.
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Carbono/química , Lauraceae/crescimento & desenvolvimento , Nitrogênio/química , Fósforo/química , Folhas de Planta/química , Ecossistema , Lauraceae/química , Fatores de TempoRESUMO
Hoodia gordonii extract contains steroid glycosides, fatty acids, plant sterols and polar organic material. Certain steroid glycosides show appetite suppressant activities following oral ingestion. This study describes the validation of a bioanalytical method for the quantification of one of the steroid glycosides, H.g.-12 (≈ 10% (w/w) of the extract), in mouse, rat, rabbit and human plasma. The method utilises a liquid-liquid extraction with methyl-tert-butyl ether followed by chromatographic separation on a 2.1 × 50 mm C(18) Genesis high performance liquid chromatography (HPLC) column and detection on a triple quadrupole mass spectrometer. Detection of H.g.-12 and its stable isotope internal standards is performed using positive TurboIonspray™ ionisation in multiple reaction monitoring mode. The validation procedure demonstrated assay sensitivity, linearity, accuracy, precision and selectivity over the calibration range of 0.5-150 ng/mL in human plasma (500 µL sample volume), 1.0-100 ng/mL in rat and rabbit plasma (150 µL sample volume) and 1.0-250 ng/mL in mouse plasma (150 µL sample volume) with good recoveries (≥ 77%). H.g.-12 was stable in plasma for ≥ 6 months at -20°C, for up to 4h at ambient temperature (ca22°C) and after 3 freeze-thaw cycles. Plasma extracts were stable for up to 24h at ambient temperature.
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Apocynaceae/química , Depressores do Apetite/química , Glicosídeos/sangue , Animais , Fracionamento Químico , Cromatografia Líquida de Alta Pressão , Glicosídeos/análise , Humanos , Limite de Detecção , Camundongos , Extratos Vegetais/sangue , Extratos Vegetais/química , Coelhos , Ratos , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em TandemRESUMO
The alkylating agent temozolomide, in combination with surgery and radiation, is the current standard of care for patients with glioblastoma. However, despite this extensive therapeutic effort, the inclusion of temozolomide extends survival only by a few short months. Among the factors contributing to chemoresistance is elevated expression of the endoplasmic reticulum (ER) chaperone GRP78 (glucose-regulated protein 78; BiP), a key pro-survival component of the ER stress response system. Because the green tea component EGCG (epigallocatechin 3-gallate) had been shown to inhibit GRP78 function, we investigated whether this polyphenolic agent would be able to increase the therapeutic efficacy of temozolomide in preclinical models of glioblastoma. Mice with intracranially implanted human U87 (p53 wild type) or U251 (p53 mutant) glioblastoma cells were treated with temozolomide and EGCG, alone and in combination. We found that EGCG alone did not provide survival benefit, but significantly improved the existing therapeutic effect of temozolomide, i.e., life extension was substantially greater under combination therapy as compared to temozolomide therapy alone. Immunohistochemical analysis of tumor tissue revealed increased expression levels of GRP78 in temozolomide-treated animals, which was diminished when temozolomide was combined with EGCG. Parallel in vitro experiments with siRNA targeting GRP78 or its major pro-apoptotic antagonist CHOP (CCAAT/enhancer binding protein homologous protein/GADD153) further established a critical role of the ER stress response system, where si-GRP78 sensitized cells to treatment with temozolomide, and si-CHOP provided protection from drug-induced toxicity. Thus, ER stress-regulatory components affect the chemotherapeutic response of glioblastoma cells to treatment with temozolomide, and inclusion of EGCG is able to increase the therapeutic efficacy of this DNA-damaging agent.
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Neoplasias Encefálicas/tratamento farmacológico , Camellia sinensis/química , Catequina/análogos & derivados , Dacarbazina/análogos & derivados , Glioblastoma/tratamento farmacológico , Animais , Antineoplásicos Alquilantes/uso terapêutico , Catequina/uso terapêutico , Linhagem Celular Tumoral , Dacarbazina/uso terapêutico , Modelos Animais de Doenças , Sinergismo Farmacológico , Chaperona BiP do Retículo Endoplasmático , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Knockout , TemozolomidaRESUMO
OBJECTIVE: To explore the effect of acupuncture-drug compound anesthesia on immune function in patients with extracorporeal circulation undergoing cardiac surgery. METHODS: Thirty cases undergoing cardiac surgery which included atrial septal defect neoplasty, ventricular septal defect neoplasty, mitral valve replacement and pulmonary valve coarctotomy were randomly divided into group A and group B, 15 cases in each group. Group A was given general anesthesia plus acupuncture at Neiguan (PC 6), Lieque (LU 7) and Yunmen (LU 2), and group B was given simple general anesthesia. Tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2) and interleukin-10 (IL-10) levels before and after surgery were compared. RESULTS: The level of TNF-alpha was increased and the levels of IL-2 and IL-10 in the serum were decreased in both groups after extracorporeal circulation for 2 h and 24 h, and the ranges of all changes were more less in group A (all P < 0.05). CONCLUSION: Compared with simple general anesthesia, acupuncture-drug compound anesthesia can improve immune suppression partially in the perioperative periods under the same conditions of controlling anesthesia degree.
Assuntos
Analgesia por Acupuntura , Anestesia Geral , Cardiopatias/imunologia , Cardiopatias/cirurgia , Mediadores da Inflamação/sangue , Adulto , Procedimentos Cirúrgicos Cardíacos , Feminino , Cardiopatias/sangue , Humanos , Interleucina-10/sangue , Interleucina-2/sangue , Masculino , Pessoa de Meia-Idade , Assistência Perioperatória , Fator de Necrose Tumoral alfa/sangue , Adulto JovemRESUMO
OBJECTIVE: To observe the effect of electroacupuncture (EA) on serum tumor necrosis factor-alpha (TNF-alpha), IL-1 and IL-4 levels in rats with chronic inflammatory pain, so as to investigate its underlying mechanism in relieving pain. Methods Forty-eight SD rats were randomized into control, model, EA-unilateral ST 36, and EA-bilateral ST 36 groups, with 12 rats in each. Chronic inflammatory pain model was established by intradermal injection of Freund's complete adjuvant (0.1 mL) into the right paw. EA (2 Hz/100 Hz, 1-2-3 mA) was applied to the unilateral "Zusanli" (ST 36) and bilateral "Zusanli" (ST 36) for 30 min, once every 3 days and continuously for 4 weeks. Serum TNF-alpha, IL-1 and IL-4 contents were detected by enzyme linked immunosorbent assay. Results Compared with the control group, serum TNF-alpha, IL-1 and IL-4 contents of the model group increased significantly (P < 0.01), while in comparison with the model group, the levels of the 3 indexes were decreased considerably in both unilateral and bilateral ST 36 groups (P < 0.01). No significant differences were found between the unilateral ST 36 and bilateral ST 36 groups in serum TNF-alpha, IL-1 and IL-4 contents (P > 0.05). CONCLUSION: EA can effectively lower serum TNF-alpha, IL-1 and IL-4 levels in inflammatory pain rats, which may contribute to its effect in relieving inflammatory pain.
Assuntos
Pontos de Acupuntura , Eletroacupuntura , Interleucina-1/sangue , Interleucina-4/sangue , Manejo da Dor , Dor/imunologia , Fator de Necrose Tumoral alfa/sangue , Animais , Doença Crônica/terapia , Modelos Animais de Doenças , Humanos , Interleucina-1/imunologia , Interleucina-4/imunologia , Masculino , Dor/sangue , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/imunologiaRESUMO
High-performance liquid chromatography (HPLC)-UV and HPLC-Mass Spectrometry (MS) methods were developed for the quantitative analysis of the family of Hoodia gordonii steroid glycosides with appetite suppressing properties in dried plant material, in purified and enriched extracts and in various prototype food-products fortified with H. gordonii extracts. For solid materials, e.g. dried plants or for non-fatty foods, extraction of the steroid glycosides is performed using methanol. For products where the steroid glycosides are present in an oil matrix, direct injection of the oil after dilution in tetrahydrofuran is applied. The HPLC separation is performed on an octyl-modified reversed-phase column in the gradient mode with UV detection at lambda = 220 nm. Quantification is performed against an external calibration line prepared using either one of the pure steroid glycosides or geranyl-tiglate. Short- and long-term repeatabilities of the methods are better than 3 and 6%, respectively. Recoveries are better than 85%, even in the analysis of the least abundant steroid glycosides in a complex yoghurt drink. Linearity is better than 3-4 orders of magnitude and the detection limits are below approximately 2 microg g(-1) for the individual steroid glycosides in dried plant material and food products. HPLC-MS is used to confirm that the steroid glycosides contain the characteristic steroid core, the carbohydrate chain and the tigloyl group.