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1.
Talanta ; 232: 122395, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34074391

RESUMO

Mycotoxins contamination in agricultural products poses a serious threat to human and animal health, so rapid and sensitive nanosensors for simultaneous determination of multiple mycotoxins in food samples are highly desirable for food safety monitoring. Herein, we report the fabrication of functional dual-colored persistent luminescence nanoparticles (PLNPs) in conjunction with Fe3O4 magnetic nanoparticles as a nanosensor for the simultaneous biosensing of aflatoxin B1 (AFB1) and zearalenone (ZEN) in food samples. Two types of PLNPs with a single excitation wavelength, Zn2GeO4:Mn2+ and Zn1.25Ga1.5Ge0.25O4:Cr3+,Yb3+,Er3+, are employed as the signal units, and aptamers with high affinity and specificity to the corresponding mycotoxins are used as the recognition units. The nanosensor was fabricated by hybridizing the aptamer modified PLNPs with the complementary DNA modified Fe3O4. The developed nanosensor offers the integrated merits of autofluorescence-free detection of persistent luminescence, the high specificity of aptamer and the high speed of magnetic separation, allowing highly sensitive and selective detection of AFB1 and ZEN in food samples with the limits of detection of 0.29 pg mL-1 for AFB1 and 0.22 pg mL-1 for ZEN and the recoveries of 93.6%-103.2% for AFB1 and 94.7%-105.1% for ZEN. This work also provides a novel universal PLNPs-based optical platform for the simultaneous detection of multiple contaminants in complex samples.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Micotoxinas , Zearalenona , Aflatoxina B1/análise , Animais , Contaminação de Alimentos/análise , Humanos , Limite de Detecção , Luminescência , Micotoxinas/análise , Zearalenona/análise
3.
Huan Jing Ke Xue ; 42(6): 2966-2974, 2021 Jun 08.
Artigo em Chinês | MEDLINE | ID: mdl-34032096

RESUMO

Migration characteristics of the heavy metals Fe, Zn, Mn, and Ni during the preparation of biochar from municipal sludge were studied, and the optimal pyrolysis temperature for the preparation of biochar was determined based on potential environmental risks. Four heavy metals (Fe, Zn, Mn, and Ni) with high total contents in the biochar were selected to determine their species and content changes under different pyrolysis temperatures using the BCR extraction method. An environmental risk assessment for sludge-based biochar was also carried out using the potential ecological risk index (PERI) and risk assessment code (RAC). The results showed that the volatility of the four metals is ranked as follows:Zn>Mn>Fe>Ni. The distribution and transformation of the four metal species were different, but their migration paths shared similar characteristics. In the pyrolysis stage at low temperatures (<500℃), unstable fractions gradually changed into more stable species; under high temperatures (>500℃), some of the oxidizable and residual fractions were broken, which transformed into reducible fractions, and other fractions escaped into the atmosphere. In the environmental risk assessment, biochar prepared under high pyrolysis temperatures (>500℃) showed lower environmental risks, with the best outcomes at 500℃.

4.
Inflammation ; 44(1): 249-260, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33098521

RESUMO

In order to investigate efficacy of FGF21 combine dexamethasone (Dex) on rheumatoid arthritis (RA) meanwhile reduce side effects of dexamethasone. We used combination therapy (Dex 15 mg/kg + FGF21 0.25 mg/kg, Dex 15 mg/kg + FGF21 0.5 mg/kg or Dex 15 mg/kg + FGF21 1 mg/kg) and monotherapy (Dex 15 mg/kg or FGF21 1 mg/kg) to treat CIA mice induced by chicken type II collagen, respectively. The effects of treatment were determined by arthritis severity score, histological damage, and cytokine production. The levels of oxidative stress parameters, liver functions, and other blood biochemical indexes were detected to determine FGF21 efficiency to side effects of dexamethasone. Oil red O was performed to detect the effects of FGF21 and dexamethasone on fat accumulation in HepG2 cells. The mechanism of FGF21 improves the side effects of dexamethasone which was analyzed by Western blotting. This combination proved to be therapeutically more effective than dexamethasone or FGF21 used singly. FGF21 regulates oxidative stress and lipid metabolism by upregulating dexamethasone-inhibited SIRT-1 and then activating downstream Nrf-2/HO-1and PGC-1. FGF21 and dexamethasone are highly effective in the treatment of arthritis; meanwhile, FGF21 may overcome the limited therapeutic response and Cushing syndrome associated with dexamethasone.


Assuntos
Anti-Inflamatórios/administração & dosagem , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Dexametasona/administração & dosagem , Fatores de Crescimento de Fibroblastos/administração & dosagem , Animais , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Galinhas , Dexametasona/efeitos adversos , Sinergismo Farmacológico , Quimioterapia Combinada , Feminino , Células Hep G2 , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Resultado do Tratamento
5.
J Asian Nat Prod Res ; 23(7): 675-680, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32869691

RESUMO

Two new selariscinins named selariscinin F (1) and selariscinin G (2), along with one known selariscinin D (3) were isolated from Selaginella tamariscina. The structures of 1-3 were elucidated on the basis of chemical and spectral analysis, including 1D, 2D NMR analyses and HRESIMS.


Assuntos
Selaginellaceae , Espectroscopia de Ressonância Magnética , Estrutura Molecular
6.
Ann Transl Med ; 8(20): 1295, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33209875

RESUMO

Background: In Chinese herbal medicine, Tanshinone IIA (Tan-IIA) is one of the main compounds extracted from Salvia miltiorrhiza Bunge. Tan-IIA has been demonstrated to inhibit the growth of various tumors. However, the detailed molecular and cellular mechanisms of the antitumor effect of Tan-IIA have yet to be fully illuminated. Methods: A2780 and ID-8 were treated with 0, 1.2, 2.4, 4.8, or 9.6 µg/mL Tan-IIA for 24 hours. Cell counting Kit-8 assay and EdU staining were used to evaluate cell proliferation. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and flow cytometry were performed to analyze apoptosis. Western blot was carried out to determine the protein levels. Flow cytometry was used for cell cycle analysis. The levels of mRNA expression were analyzed by real-time polymerase chain reaction. The anti-tumor effect of Tan-IIA was observed in a tumor-bearing mouse model. Results: Tan-IIA inhibited the proliferation of ovarian cancer cells in a dose-dependent manner by inducing G2/M phase arrest. It also down-regulated B-cell lymphoma 2 (Bcl-2) and up-regulated Bcl-2-associated X protein (Bax) in ovarian cancer cells to induce apoptosis, and suppressed cell migration by inhibiting focal adhesion kinase phosphorylation. Tan-IIA significantly reduced vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX2) mRNA expression in ovarian cancer cells. In vivo, Tan-IIA significantly inhibited tumor growth by inducing apoptosis and promoting anti-angiogenesis. Conclusions: The results of this study shed light on the molecular and cellular mechanisms for the antitumor effect of Tan-IIA.

7.
Genome Biol ; 21(1): 257, 2020 10 06.
Artigo em Inglês | MEDLINE | ID: mdl-33023639

RESUMO

Prime editing is a novel and universal CRISPR/Cas-derived precision genome-editing technology that has been recently developed. However, low efficiency of prime editing has been shown in transgenic rice lines. We hypothesize that enhancing pegRNA expression could improve prime-editing efficiency. In this report, we describe two strategies for enhancing pegRNA expression. We construct a prime editing vector harboring two pegRNA variants for W542L and S621I double mutations in ZmALS1 and ZmALS2. Compared with previous reports in rice, we achieve much higher prime-editing efficiency in maize. Our results are inspiring and provide a direction for the optimization of plant prime editors.


Assuntos
Acetolactato Sintase/genética , Edição de Genes/métodos , Mutagênese Sítio-Dirigida/métodos , RNA Guia/metabolismo , Zea mays/genética , Edição de Genes/estatística & dados numéricos , Vetores Genéticos , Plantas Geneticamente Modificadas , RNA Guia/genética , Zea mays/enzimologia
8.
Sci Rep ; 10(1): 16943, 2020 10 09.
Artigo em Inglês | MEDLINE | ID: mdl-33037234

RESUMO

Mutations that lead to constitutive activation of key regulators in cellular processes are one of the most important drivers behind vigorous growth of cancer cells, and are thus prime targets in cancer treatment. BRAF V600E mutation transduces strong growth and survival signals for cancer cells, and is widely present in various types of cancers including lung cancer. A combination of BRAF inhibitor (dabrafenib) and MEK inhibitor (trametinib) has recently been approved and significantly improved the survival of patients with advanced NSCLC harboring BRAF V600E/K mutation. To improve the detection of BRAF V600E/K mutation and investigate the incidence and clinicopathological features of the mutation in lung cancer patients of southern Taiwan, a highly sensitive and specific real-time quantitative PCR (RT-qPCR) method, able to detect single-digit copies of mutant DNA, was established and compared with BRAF V600E-specific immunohistochemistry. Results showed that the BRAF V600E mutation was present at low frequency (0.65%, 2/306) in the studied patient group, and the detection sensitivity and specificity of the new RT-qPCR and V600E-specific immunohistochemistry both reached 100% and 97.6%, respectively. Screening the BRAF V600E/K mutation with the RT-qPCR and V600E-specific immunohistochemistry simultaneously could help improve detection accuracy.


Assuntos
Neoplasias Pulmonares/genética , Mutação/genética , Proteínas Proto-Oncogênicas B-raf/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Idoso , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Humanos , Imidazóis/uso terapêutico , Imuno-Histoquímica/métodos , Neoplasias Pulmonares/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Oximas/uso terapêutico , Inibidores de Proteínas Quinases/uso terapêutico , Piridonas/uso terapêutico , Pirimidinonas/uso terapêutico , Sensibilidade e Especificidade , Taiwan
9.
Talanta ; 218: 121101, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32797868

RESUMO

Serious ochratoxin A (OTA) contamination necessitates the development of rapid, sensitive and selective analytical methods for its determination in food safety. Herein, we report a persistent luminescence resonance energy transfer (LRET) based aptasensor for the autofluorescence-free detection of OTA. OTA aptamer functionalized persistent luminescence nanorod (PLNR) Zn2GeO4:Mn2+ and the aptamer complementary DNA modified gold nanoparticle (AuNP) were used as the donor and the acceptor, respectively. The developed LRET aptasensor integrated the advantages of the long-lasting persistent luminescence of PLNR, the high selectivity of aptamer and the low probe background of LRET sensors, allowing autofluorescence-free detection of OTA in biological samples with high sensitivity and selectivity. The developed LRET aptasensor gave an excellent linearity in the range of 0.01-10 ng mL-1, the detection limit of 3 pg mL-1 and the precision of 2.7% (RSD, n = 11) at 1 ng mL-1 level. The applicability of the developed aptasensor was demonstrated by analyzing beer samples for OTA with the recoveries of 92.3%-104%.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Micotoxinas , Nanotubos , Ocratoxinas , Ouro , Limite de Detecção , Luminescência , Ocratoxinas/análise
10.
Front Psychiatry ; 11: 608, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32733289

RESUMO

Background: Depressive symptoms are common in empty-nest elderly in China, but the reported prevalence rates across studies are mixed. This is a meta-analysis of the pooled prevalence of depressive symptoms (depression hereafter) in empty-nest elderly in China. Methods: Two investigators independently conducted a systematic literature search in both English (PubMed, EMBASE, PsycINFO, Web of Science, and Cochrane Library) and Chinese (CNKI and Wan Fang) databases. Data were analyzed using the Comprehensive Meta-Analysis program. Results: A total of 46 studies with 36,791 subjects were included. The pooled prevalence of depression was 38.6% (95%CI: 31.5-46.3%). Compared with non-empty-nest elderly, empty-nest elderly were more likely to suffer from depression (OR=2.0, 95%CI: 1.4 to 2.8, P<0.001). Subgroup and meta-regression analyses revealed that mild depression were more common in empty-nest elderly than moderate or severe depression (P<0.001). In addition, living alone (P=0.002), higher male proportion (ß=0.04, P<0.001), later year of publication (ß=0.09, P<0.001) and higher study quality score (ß=0.62, P<0.001) were significantly associated with higher prevalence of depression. Conclusion: In this meta-analysis, the prevalence of depression in empty-nest elderly was high in China. Considering the negative impact of depression on health outcomes and well-being, regular screening and appropriate interventions need to be delivered for this vulnerable segment of the population.

11.
Sci Rep ; 10(1): 12266, 2020 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-32703962

RESUMO

Poor sleep quality is associated with negative health outcomes and high treatment burden. This study investigated the prevalence of poor sleep quality and its socio-demographic correlates among older adults in Hebei province, which is a predominantly agricultural region of China. A large-scale cross-sectional epidemiological survey was conducted from April to August 2016. The study used a multistage, stratified, cluster random sampling method. Sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI). A total of 3,911 participants were included. The prevalence of poor sleep quality (defined as PSQI > 7) was 21.0% (95% CI 19.7-22.2%), with 22.3% (95% CI 20.9-23.8%) in rural areas and 15.9% (95% CI 13.4-18.4%) in urban areas. Multivariable logistic regression analyses found that female gender (P < 0.001, OR 2.4, 95% CI 2.00-2.82), rural areas (P = 0.002, OR 1.5, 95% CI 1.14-1.86), presence of major medical conditions (P < 0.001, OR 2.4, 95% CI 2.02-2.96) and family history of psychiatric disorders (P < 0.001, OR 2.7, 95% CI 1.60-4.39) were independently associated with higher risk of poor sleep quality. Poor sleep quality was common among older adults in Hebei province of China. Regular assessment of sleep quality and accessible sleep treatments for older population should be provided in agricultural areas of China.


Assuntos
Qualidade de Vida , Transtornos do Sono-Vigília/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Razão de Chances , Prevalência , Vigilância em Saúde Pública , Fatores de Risco , Fatores Sexuais , Fatores Socioeconômicos
12.
Ann Transl Med ; 8(12): 752, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32647677

RESUMO

Background: Paclitaxel is a widely used clinical first line chemotherapy drug for ovarian carcinoma. Tanshinone I (Tan-I) is one of the vital fat-soluble components, which derived from Chinese herbal medicine, Salvia miltiorrhiza Bunge. Herein, we evaluated whether Tan-I could enhance the efficacy of ovarian cancer to chemotherapy of Paclitaxel. Methods: Ovarian cancer cells A2780 and ID-8 were exposed with Tan-I (4.8 µg/mL), Paclitaxel (0.1 µg/mL), or Tan-I combination with Paclitaxel for 24 hours. The cell proliferation was analyzed by CCK8 and EdU staining. Cell apoptosis was analyzed by the TUNEL assay and flow cytometry. The protein levels were determined by western blot. Cell migration was analyzed by Transwell and wound healing. Cell senescence was analyzed by senescence-associated b-galactosidase staining. Antitumor activity was analyzed by a subcutaneous tumor xenograft model of human ovarian cancer in nude mice. The protein expression and apoptosis level of tumor tissues were analyzed by immunohistochemistry and TUNEL staining. Results: Tan-I treatment significantly elevated the Paclitaxel-cause reduction of A2780 and ID-8 cell proliferation and cell migration. Tan-I combination with Paclitaxel promotes apoptosis of cancer cells by promoting Bax expression and Bcl-2 expression. Besides, Tan-I treatment can notably increase Paclitaxel-inducing cell senescence by promoting DNA damage and senescence-associated proteins such as p21 and p16. Furthermore, the result of the transplanted tumor model indicated that Tan-I combination with Paclitaxel could inhibit tumor growth in vivo by inhibiting cell proliferation and inducing cell apoptosis. Conclusions: Natural compound Tan-I enhances the efficacy of ovarian cancer to Paclitaxel chemotherapy. The results will help to supply the potential clinical use of ovarian carcinoma cells.

13.
Int J Biol Macromol ; 157: 434-443, 2020 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-32315678

RESUMO

The fractional polysaccharide SMWP-U&E was isolated from Salvia miltiorrhiza residue. SMWP-U&E consists of 91.40% carbohydrates and has an average molecular weight of 5.07 × 105 Da. The polysaccharides are mainly composed of arabinose (Ara), fructose (Fru), mannose (Man), glucose (Glc), and galactose (Gal), and their mole percentages are 3.72%, 4.11%, 6.18%, 32.08% and 53.91%, respectively. When effected on weaned piglets, 1.5 g/kg SMWP-U&E supplementation significantly increased the villus height to crypt depth ratio in ileum. PCR-denaturing gradient gel electrophoresis and qRT-PCR results indicated that SMWP-U&E supplementation could change the density of intestinal microbiota and the populations of Lactobacillus and Escherichia coli in jejunum, ileum, caecum, and colon. The supplementation also increased contents of IgA, IgG, IgM, IL-2, IFN-γ, and IL-10; promoted T-AOC and SOD activities; and reduced MDA level in the serum. These findings suggest that SMWP-U&E improves digestion and nutrient absorption in weaned piglets, exerts beneficial effects on intestinal morphology and microflora, and enhances the immune and antioxidant capabilities in mode of weaned piglets. Thus, SMWP-U&E exhibits potential as a new type of plant-derived additive and novel prebiotics.


Assuntos
Extratos Vegetais/química , Extratos Vegetais/farmacologia , Polissacarídeos/química , Polissacarídeos/farmacologia , Salvia miltiorrhiza/química , Animais , Biomarcadores , Microbioma Gastrointestinal/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Monossacarídeos/química , Extratos Vegetais/isolamento & purificação , Polissacarídeos/isolamento & purificação , Espectroscopia de Infravermelho com Transformada de Fourier , Relação Estrutura-Atividade , Suínos
14.
Korean J Orthod ; 50(2): 86-97, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32257934

RESUMO

Objective: To propose a three-dimensional (3D) method for evaluating temporomandibular joint (TMJ) changes during Twin-block treatment. Methods: Seventeen patients with Class II division 1 malocclusion treated using Twin-block and nine untreated patients with a similar malocclusion were included in this research. We collected their cone beam computed tomography (CBCT) data from before and 8 months after treatment. Segmentations were constructed using ITK-SNAP. Condylar volume and superficial area were measured using 3D Slicer. The 3D landmarks were identified on CBCT images by using Dolphin software to assess the condylar positional relationship. 3D models of the mandible and glenoid fossa of the patients were constructed and registered via voxel-based superimposition using 3D Slicer. Thereafter, skeletal changes could be visualized using 3DMeshMetric in any direction of the superimposition on a color-coded map. All the superimpositions were measured using the same scale on the distance color-coded map, in which red color represents overgrowth and blue color represents resorption. Results: Significant differences were observed in condylar volume, superficial area, and condylar position in both groups after 8 months. Compared with the control group (CG), the Twin-block group exhibited more obvious condyle-fossa modifications and joint positional changes. Moreover, on the color-coded map, more obvious condyle-fossa modifications could be observed in the posterior and superior directions in the Twin-block group than in the CG. Conclusions: We successfully established a 3D method for measuring and evaluating TMJ changes caused by Twin-block treatment. The treatment produced a larger condylar size and caused condylar positional changes.

15.
J Chromatogr A ; 1620: 461036, 2020 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-32201039

RESUMO

Leaves, flowers, fruits and stems (44 sample groups) were collected from mature Camptotheca acuminate during 2017.3-2018.3 and classified by ultra-high performance liquid chromatography coupled with quadrupole-time of flight-mass spectrometry based metabolomics. One hundred metabolites including forty-seven alkaloids, fifteen terpenes, thirty-two polyphenols and six other metabolites were rapidly identified through the in-house database alignment at first glance. Thirty-three alkaloids classified into five groups including camptothecin group (CG1-13), pumiloside group (PG1-5), strictosidinic acid group (SG1-3), vincosamide group (VG1-7), and a new hybrid group, vincosamide-camptothecin group (VC1-5) were mined and further characterized by MS/MS analyses. The identification of two untapped biosynthetic precursors, 2-hydroxypumiloside (PG2) and 16­hydroxy­15, 16-dihydrocamptothecoside (CG3), along with sixteen new alkaloids enables us for a better understanding of camptothecin biogenetic reasoning. The underlying enzymes involved in camptothecin biosynthesis were also proposed according to the guiding metabolic map, thus purposefully mining of enzymes involved in the downstream biosynthetic pathway of camptothecin could be initiated with the help of this map.


Assuntos
Alcaloides/análise , Vias Biossintéticas , Camptotheca/química , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas em Tandem/métodos , Camptotecina/análogos & derivados , Camptotecina/análise , Camptotecina/química , Camptotecina/metabolismo , Carbolinas/análise , Carbolinas/química , Bases de Dados como Assunto , Análise Discriminante , Glicosídeos/análise , Glicosídeos/química , Alcaloides Indólicos/análise , Alcaloides Indólicos/química , Análise dos Mínimos Quadrados , Redes e Vias Metabólicas , Metaboloma , Metabolômica , Análise Multivariada , Análise de Componente Principal
16.
Nat Prod Res ; 34(22): 3285-3288, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30931646

RESUMO

In this study, the chemical composition, antimicrobial and antioxidant activities of Litsea cubeba essential oils extracted in different months were analysed. Results showed that the essential oil contents of fruits collected in June, July and August were 3.47%, 5.02% and 5.64%, respectively, and contained 13, 17 and 17 components, respectively. Neral and geranial were the main components and accounted for 54.76%. The essential oil extracted from fruits collected in July had the highest antimicrobial activity against Staphylococcus aureus, Escherichia coli, and Salmonella typhimurium, and it was the most effective based on the OH· scavenging activity test. The essential oil extracted from fruits collected in August was the most effective based on the test for DPPH· scavenging activity and ferric reducing antioxidant power. Considering the contents, chemical compositions and antimicrobial and antioxidant activities, the appropriate harvest time for L. cubeba essential oils is from July to August.


Assuntos
Antibacterianos/farmacologia , Antioxidantes/farmacologia , Litsea/química , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Monoterpenos Acíclicos/análise , Antibacterianos/química , Antioxidantes/química , China , Escherichia coli/efeitos dos fármacos , Frutas/química , Testes de Sensibilidade Microbiana , Salmonella typhimurium/efeitos dos fármacos , Fatores de Tempo
17.
Cell Prolif ; 53(2): e12739, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31820522

RESUMO

OBJECTIVES: Tanshinone I (Tan-I) is one of the vital fatsoluble monomer components, which extracted from Chinese medicinal herb Salvia miltiorrhiza Bunge. It has been shown that Tan-I exhibited anti-tumour activities on different types of cancers. However, the underlying mechanisms by which Tan-Ⅰ regulates apoptosis and autophagy in ovarian cancer remain unclear. Thus, this study aimed to access the therapy effect of Tan-Ⅰ and the underlying mechanisms. METHODS: Ovarian cancer cells A2780 and ID-8 were treated with different concentrations of Tan-Ⅰ (0, 1.2, 2.4, 4.8 and 9.6 µg/mL) for 24 hours. The cell proliferation was analysed by CCK8 assay, EdU staining and clone formation assay. Apoptosis was assessed by the TUNEL assay and flow cytometry. The protein levels of apoptosis protein (Caspase-3), autophagy protein (Beclin1, ATG7, p62 and LC3II/LC3I) and PI3K/AKT/mTOR pathway were determined by Western blot. Autophagic vacuoles in cells were observed with LC3 dyeing using confocal fluorescent microscopy. Anti-tumour activity of Tan-Ⅰ was accessed by subcutaneous xeno-transplanted tumour model of human ovarian cancer in nude mice. The Ki67, Caspase-3 level and apoptosis level were analysed by immunohistochemistry and TUNEL staining. RESULTS: Tan-Ⅰ inhibited the proliferation of ovarian cancer cells A2780 and ID-8 in a dose-dependent manner, based on CCK8 assay, EdU staining and clone formation assay. In additional, Tan-Ⅰ induced cancer cell apoptosis and autophagy in a dose-dependent manner in ovarian cancer cells by TUNEL assay, flow cytometry and Western blot. Tan-Ⅰ significantly inhibited tumour growth by inducing cell apoptosis and autophagy. Mechanistically, Tan-Ⅰ activated apoptosis-associated protein Caspase-3 cleavage to promote cell apoptosis and inhibited PI3K/AKT/mTOR pathway to induce autophagy. CONCLUSIONS: This is the first evidence that Tan-Ⅰ induced apoptosis and promoted autophagy via the inactivation of PI3K/AKT/mTOR pathway on ovarian cancer and further inhibited tumour growth, which might be considered as effective strategy.


Assuntos
Abietanos/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Neoplasias Ovarianas/tratamento farmacológico , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Animais , Antineoplásicos Fitogênicos/farmacologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias Ovarianas/metabolismo , Transdução de Sinais/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
18.
Plant Physiol ; 181(4): 1441-1448, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31558579

RESUMO

The lack of efficient delivery methods is a major barrier to clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas)-mediated genome editing in many plant species. Combinations of morphogenic regulator (MR) genes and ternary vector systems are promising solutions to this problem. In this study, we first demonstrated that MR vectors greatly enhance maize (Zea mays) transformation. We then tested a CRISPR/Cas9 MR vector in maize and found that the MR and CRISPR/Cas9 modules have no negative influence on each other. Finally, we developed a novel ternary vector system to integrate the MR and CRISPR/Cas modules. Our ternary vector system is composed of new pGreen-like binary vectors, here named pGreen3, and a pVS1-based virulence helper plasmid, which also functions as a replication helper for the pGreen3 vectors in Agrobacterium tumefaciens The pGreen3 vectors were derived from the plasmid pRK2 and display advantages over pGreen2 vectors regarding both compatibility and stability. We demonstrated that the union of our ternary vector system with MR gene modules has additive effects in enhancing maize transformation and that this enhancement is especially evident in the transformation of recalcitrant maize inbred lines. Collectively, our ternary vector system-based tools provide a user-friendly solution to the low efficiency of CRISPR/Cas delivery in maize and represent a basic platform for developing efficient delivery tools to use in other plant species recalcitrant to transformation.


Assuntos
Sistemas CRISPR-Cas/genética , Genes de Plantas , Vetores Genéticos/genética , Morfogênese/genética , Zea mays/crescimento & desenvolvimento , Zea mays/genética , Agrobacterium tumefaciens/genética , Transformação Genética
19.
PLoS One ; 14(8): e0220670, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31369639

RESUMO

Somatic mutations of MET gene are emerging as important driver mutations for lung cancers. To identify the common clinicopathological features of MET exon 14 skipping mutations and amplification and clarify whether the two MET gene alterations cause protein overexpression were investigated using 196 lung cancer samples of Taiwan through real time-qPCR/sequencing, fluorescence in situ hybridization, and immunohistochemistry. The two MET gene alterations are both present in low frequency, ~1%, in the studied lung cancer population of Taiwan. MET exon 14 skipping mutations were identified from two early-stage patients, who were both relatively advanced in age, and did not carry other driver mutations. One was an adenocarcinoma and the other was a rare carcinosarcoma. Three gene amplifications cases were identified. Neither of the two MET gene alterations would lead to protein overexpression; hence, direct detection in nucleic acid level would be a preferred and straightforward solution for the identification of skipping mutations. The presence of MET exon 14 mutations in minor histological types of lung cancers urge to extend screening scope of this mutation in lung cancer and treatment response evaluation in clinical trials. These would be important next steps for the success of MET target therapy in clinical practice.


Assuntos
Éxons/genética , Amplificação de Genes/genética , Neoplasias Pulmonares/genética , Mutação/genética , Proteínas Proto-Oncogênicas c-met/genética , Adenocarcinoma/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinossarcoma/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Hibridização in Situ Fluorescente , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Taiwan
20.
Cancers (Basel) ; 11(6)2019 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-31185703

RESUMO

Mutations in the epidermal growth factor receptor (EGFR) are associated with various solid tumors. This study aimed to compare two methods for the detection of EGFR mutations in circulating tumor DNA (ctDNA) from lung adenocarcinoma (LUAD) patients and to evaluate the clinical significance of EGFR mutations in ctDNA. In this prospective cohort study, the EGFR mutation status of 77 patients with stage IIIB or IV LUAD was first determined using lung cancer tissue. The amplification refractory mutation system (ARMS) and single allele base extension reaction combined with mass spectroscopy (SABER/MassARRAY) methods were also used to detect EGFR mutations in plasma ctDNA from these patients and then compared using the EGFR mutation status in lung cancer tissue as a standard. Furthermore, the relationship between the presence of EGFR mutations in ctDNA after receiving first-line EGFR-tyrosine kinase inhibitor (EGFR-TKI) therapy and survival was evaluated. The overall sensitivity and specificity for the detection of EGFR mutations in plasma ctDNA by ARMS and SABER/MassARRAY were 49.1% vs. 56% and 90% vs. 95%, respectively. The agreement level between these methods was very high, with a kappa-value of 0.88 (95% CI 0.77-0.99). Moreover, 43 of the patients who carried EGFR mutations also received first-line EGFR-TKI therapy. Notably, patients with EGFR mutations in plasma ctDNA had significantly shorter progression-free survival (9.0 months, 95% CI 7.0-11.8, vs. 15.0 months, 95% CI 11.7-28.2; p = 0.02) and overall survival (30.6 months, 95% CI 12.4-37.2, vs. 55.6 months, 95% CI 25.8-61.8; p = 0.03) compared to those without detectable EGFR mutations. The detection of EGFR mutations in plasma ctDNA is a promising, minimally invasive, and reliable alternative to tumor biopsy, and the presence of EGFR mutations in plasma ctDNA after first-line EGFR-TKI therapy is associated with poor prognosis.

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