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In nature, biological nitrogen fixation is accomplished through the π-back-bonding mechanism of nitrogenase, which poses significant challenges for mimic artificial systems, thanks to the activation barrier associated with the N≡N bond. Consequently, this motivates us to develop efficient and reusable photocatalysts for artificial nitrogen fixation under mild conditions. We employ a charge-assisted self-assembly process toward encapsulating one polyoxometalate (POM) within a dehydrated Zr-based metal-organic framework (d-UiO-66) exhibiting nitrogen photofixation activities, thereby constructing an enzyme-mimicking photocatalyst. The dehydration of d-UiO-66 is favorable for facilitating nitrogen chemisorption and activation via the unpaired d-orbital electron at the [Zr6O6] cluster. The incorporation of POM guests enhanced the charge separation in the composites, thereby facilitating the transfer of photoexcited electrons into the π* antibonding orbital of chemisorbed N2 for efficient nitrogen fixation. Simultaneously, the catalytic efficiency of SiW9Fe3@d-UiO-66 is enhanced by 9.0 times compared to that of d-UiO-66. Moreover, SiW9Fe3@d-UiO-66 exhibits an apparent quantum efficiency (AQE) of 0.254% at 550 nm. The tactics of "working-in-tandem" achieved by POMs and d-UiO-66 are extremely vital for enhancing artificial ammonia synthesis. This study presents a paradigm for the development of an efficient artificial catalyst for nitrogen photofixation, aiming to mimic the process of biological nitrogen fixation.
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DDB1-Cullin-4-associated factor-2 (DCAF2, also known as DTL or CDT2), a conserved substrate recognition protein of Cullin-RING E3 ligase 4 (CRL4), recognizes and degrades several substrate proteins during the S phase to maintain cell cycle progression and genome stability. Dcaf2 mainly expressed in germ cells of human and mouse. Our study found that Dcaf2 was expressed in mouse spermatogonia and spermatocyte. The depletion of Dcaf2 in germ cells by crossing Dcaf2fl/fl mice with stimulated by retinoic acid gene 8(Stra8)-Cre mice caused a reduction in progenitor spermatogonia and differentiating spermatogonia, eventually leading to the failure of meiosis initiation and male infertility. Further studies showed that depletion of Dcaf2 in germ cells caused abnormal accumulation of the substrate proteins, cyclin-dependent kinase inhibitor 1A (p21) and thymine DNA glycosylase (TDG), decreasing of cell proliferation, increasing of DNA damage and apoptosis. Overexpression of p21 or TDG attenuates proliferation and increases DNA damage and apoptosis in GC-1 cells, which is exacerbated by co-overexpression of p21 and TDG. The findings indicate that DCAF2 maintains the proliferation and differentiation of progenitor spermatogonia by targeting the substrate proteins p21 and TDG during the S phase.
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Background: Aged women and premature ovarian insufficiency (POI) patients have residual dormant primordial follicles that are hard to be activated through a physiological process. However, there are no effective and safe drugs to help them. Methods: We used the in vitro culture model of newborn mouse ovaries to identify the drugs that promote primordial follicle activation and study its mechanisms. It was verified by in vivo injection model of newborn mice and in vitro culture model of human ovarian tissue. In addition, we used the aged mice as a low infertility model to verify the effects of primordial follicle activation, and fertility by drugs. Results: Eleven metallic compounds activated mouse primordial follicles, and the five most effective compounds were selected for further study. Thapsigargin (TG), CrCl3, MnCl2, FeCl3 and ZnSO4 increased the levels of the glycolysis-related proteins (glucose transporter type 4, GLUT4; hexokinase 1, HK1; pyruvate kinase M2, PKM2; phosphofructokinase, liver type, PFKL), phosphorylated mammalian target of rapamycin (p-mTOR) in cultured mouse ovaries. The compound-promoted p-mTOR levels could be completely blocked by 2-DG (the inhibitor of glycolysis). The compounds also increased the levels of phosphorylated protein kinase B (p-Akt). TG-, CrCl3- and FeCl3-promoted p-Akt levels, but not MnCl2- and ZnSO4- promoted p-Akt levels, could be completely blocked by ISCK03 (the inhibitor of proto-oncogenic receptor tyrosine kinase, KIT). The injection of newborn mice with the compounds also activated primordial follicles and increased the levels of the glycolysis-related proteins, p-mTOR, and p-Akt. The oral administration of the compounds in adolescent and aged mice promoted primordial follicle activation, and had no obvious side effect. Importantly, ZnSO4 also increased ovulated oocytes, oocyte quality and offspring in aged mice. Furthermore, the compounds promoted human primordial follicle activation and increased the levels of the glycolysis-related proteins, p-mTOR, and p-Akt. Conclusion: The metallic compounds activate primordial follicles through the glycolysis-dependent mTOR pathway and/or the PI3K/Akt pathway, and the oral administration of ZnSO4 enhances fertility in aged mice. We suggest that these metallic compounds may be oral drugs to ameliorate fertility deficits in aged women and POI patients.
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Insuficiência Ovariana Primária , Proteínas Proto-Oncogênicas c-akt , Animais , Humanos , Feminino , Camundongos , Adolescente , Idoso , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Fosfatidilinositol 3-Quinases/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Insuficiência Ovariana Primária/tratamento farmacológico , Fertilidade , Mamíferos/metabolismoRESUMO
A tomato (Lycopersicon esculentum Mill.) monodehydroascorbate reductase gene (LeMDAR) was isolated. The LeMDAR-green fluorescence protein (GFP) fusion protein was targeted to chloroplast in Arabidopsis mesophyll protoplast. RNA and protein gel blot analyses confirmed that the sense- and antisense- LeMDAR were integrated into the tomato genome. The MDAR activities and the levels of reduced ascorbate (AsA) were markedly increased in sense transgenic lines and decreased in antisense transgenic lines compared with wild-type (WT) plants. Under low and high temperature stresses, the sense transgenic plants showed lower level of hydrogen peroxide (H(2)O(2)), lower thiobarbituric acid reactive substance (TBARS) content, higher net photosynthetic rate (P(n)), higher maximal photochemical efficiency of PSII (F(v)/F(m)) and fresh weight compared with WT plants. The oxidizable P700 decreased more obviously in WT and antisense plants than that in sense plants at chilling temperature under low irradiance. Furthermore, the sense transgenic plants exhibited significantly lower H(2)O(2) level, higher ascorbate peroxidase (APX) activity, greater P(n) and F(v)/F(m) under methyl viologen (MV)-mediated oxidative stresses. These results indicated that overexpression of chloroplastic MDAR played an important role in alleviating photoinhibition of PSI and PSII and enhancing the tolerance to various abiotic stresses by elevating AsA level.
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NADH NADPH Oxirredutases/metabolismo , Estresse Oxidativo , Paraquat/farmacologia , Solanum lycopersicum/enzimologia , Temperatura , Sequência de Aminoácidos , Ascorbato Peroxidases , Ácido Ascórbico/metabolismo , Cloroplastos/enzimologia , Regulação da Expressão Gênica de Plantas , Peróxido de Hidrogênio/metabolismo , Solanum lycopersicum/genética , Dados de Sequência Molecular , NADH NADPH Oxirredutases/genética , Peroxidases/metabolismo , Fotossíntese , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , RNA de Plantas/genética , Alinhamento de Sequência , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
Paraquat (PQ) is a widely used non-selective and oxidizing herbicide in farmland, orchards, flower nursery, and grassland. Overuse of PQ will accumulate in the body and affect the reproduction in mammals. In this study, we found that PQ could reduce the female fertility by oral administration for 21 days in mice. PQ exposure could impair the nuclear maturation by perturbing the spindle assembly and kinetochore-microtubule attachment to cause the misaligned chromosomes during meiosis. In the meantime, PQ exposure disturbed the mitochondrial distribution and enhanced the level of reactive oxygen species and early apoptosis, which thereby deteriorated the early embryo development. Also, PQ administration could cause some changes in epigenetic modifications such as the level of H3K9me2 and H3K27me3. Therefore, PQ administration reduces the female fertility by impairing the nuclear and cytoplasmic maturation of oocytes in mice.
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After ovulation, mammalian oocytes will undergo a time-dependent process of aging if they are not fertilized. This postovulatory aging (POA) seriously affects the oocyte quality and then impairs the subsequent fertilization and early embryo development, which should be avoided especially in assisted reproductive technology (ART). Resveratrol is an antioxidant substance that can scavenge free radicals and is effective in improving ovary functions. Here, mouse oocytes were used to investigate the effects and mechanisms of resveratrol on POA oocytes in vitro. With 1.0 µM resveratrol treatment during aging process, the rates of fertilization and blastocyst in POA oocytes increased significantly compared with those in the POA group. Resveratrol can reduce the loss of sperm binding sites by stabilizing Juno. Resveratrol can maintain the normal morphology of spindle and mitochondrion distribution and alleviate the levels of ROS and early apoptosis. Additionally, resveratrol can reduce the changes of H3K9me2. Therefore, resveratrol can significantly improve the quality of POA oocytes in vitro to enhance the rates of fertilization and blastocyst, which may be very helpful during the ART process.
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Antioxidantes/farmacologia , Fertilização in vitro/efeitos dos fármacos , Oócitos/citologia , Resveratrol/farmacologia , Animais , Células Cultivadas , Senescência Celular/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Histonas/metabolismo , Técnicas In Vitro , Fase Luteal , Camundongos , Oócitos/efeitos dos fármacos , Receptores de Superfície Celular/metabolismoRESUMO
The aim of the present study was to investigate the effects of phosphorylatable nucleus localization signal linked nucleic kinase substrate short peptide (pNNS)-conjugated chitosan (pNNS-CS) mediated miR-140 and IGF-1 in both rabbit chondrocytes and cartilage defects model. pNNS-CS was combined with pBudCE4.1-IGF-1, pBudCE4.1-miR-140, and negative control pBudCE4.1 to form pDNA/pNNS-CS complexes. Then these complexes were transfected into chondrocytes or injected intra-articularly into the knee joints. High levels of IGF-1 and miR-140 expression were detected both in vitro and in vivo. Compared with pBudCE4.1 group, in vitro, the transgenic groups significantly promoted chondrocyte proliferation, increased glycosaminoglycan (GAG) synthesis, and ACAN, COL2A1, and TIMP-1 levels, and reduced the levels of nitric oxide (NO), MMP-13, and ADAMTS-5. In vivo, the exogenous genes enhanced COL2A1, ACAN, and TIMP-1 expression in cartilage and reduced cartilage Mankin score and the contents of NO, IL-1ß, TNF-α, and GAG contents in synovial fluid of rabbits, MMP-13, ADAMTS-5, COL1A2, and COL10A1 levels in cartilage. Double gene combination showed better results than single gene. This study indicate that pNNS-CS is a better gene delivery vehicle in gene therapy for cartilage defects and that miR-140 combination IGF-1 transfection has better biologic effects on cartilage defects.
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Doenças das Cartilagens/tratamento farmacológico , Cartilagem Articular/efeitos dos fármacos , Quitosana/farmacologia , Condrócitos/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/metabolismo , MicroRNAs/metabolismo , Peptídeos/farmacologia , Animais , Doenças das Cartilagens/metabolismo , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Técnicas de Transferência de Genes , Humanos , Articulação do Joelho/metabolismo , Metaloproteinase 13 da Matriz/metabolismo , Óxido Nítrico/metabolismo , Coelhos , Líquido Sinovial/efeitos dos fármacos , Líquido Sinovial/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Transfecção/métodosRESUMO
Characterization of phosphorus (P) and its risk of loss in urban soils in Kaifeng City, Henan Province were studied through field sampling and laboratory experiments. The spatial distribution of P and the map of risk of loss were obtained using geostatistical and spatial analysis techniques. The P content in urban soils ranged from 400 to 1427 mg·kg-1, the proportions of inorganic P in total P ranged from 65% to 99%, and Olsen-P and CaCl2-P in soils were 3.41-115.03 mg·kg-1 and 0.01-9.40 mg·kg-1, respectively. The composition of P was consistent in different urban areas and P concentrations were higher in residential areas. Spatial variations in P concentrations in soils were significant; the concentrations of P in eastern Kaifeng City were higher than those in western Kaifeng and the highest concentrations were detected in central Kaifeng. Olsen-P can be used as an indicator of the leaching risk of soil P. The critical value of leaching P from the soil was 22.18 mg·kg-1 and the concentration of Olsen-P in 33.64% of urban soil samples exceeded the critical value. The highest risk of P loss existed in central Kaifeng City.
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AIM: To investigate the effect of dopamine on bone morphogenesis protein-2 (BMP-2) expression in retinal pigment epithelium (RPE) cells in vitro. METHODS: ARPE-19 cells as a human RPE cell line were cultured with dopamine for different times (2, 4, 6, 8, 12, 16 and 24h) or with different concentrations (0.1, 1, 2, 5, 10, 20, and 100 µg/mL) in vitro. BMP-2 mRNA expression level in ARPE-19 cells was analyzed with real-time polymerase chain reaction (PCR) analysis and BMP-2 protein level was measured with Western blot analysis. The active form of BMP-2 in the culture medium was measured with enzyme-linked immunosorbent assay (ELISA). RESULTS: The expression level of BMP-2 increased significantly cultured with 20 µg/mL dopamine, at different time points (P<0.05). BMP-2 mRNA level peaked 2h and the protein level peaked at 6 and 8h after treatment. The concentrations of secreted BMP-2 elevated at 12h and peaked at 24h (P<0.05) in a time-dependent manner. Treated with 100 µg/mL dopamine for 6h, the expression levels of BMP-2 mRNA and protein in ARPE-19 cells were enhanced significantly compared to that in the untreated cells (P<0.05). And secreted BMP-2 protein in the cell culture supernatant was also increased (P<0.05). CONCLUSION: Dopamine up-regulate BMP-2 expression in RPE cells, and this may be associated with its inhibitive effect on myopia development.
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Ninety-nine topsoil (0-15 cm) samples were collected from Kaifeng City, China using the grid method, and then the concentrations of As, Cd, Cr, Cu, Ni, Pb and Zn in the samples were measured by standard methods. Soil pollution levels and potential ecological risks of the heavy metals were assessed using the pollution load index (PLI) and potential ecological risk index (RI), respectively. Ordinary Kriging interpolation technique was employed to investigate the spatial distribution of PLI and RI of the city. The results showed that high pollution of Cd occurred in Kaifeng urban soils, and there was moderate pollution of Zn, slight pollution of Pb and Cu, and no pollution of Ni, Cr and As. Very high ecological risk was posed by Cd and low risk by other metals. The mean PLI of the 7 metals from all sample points was 2.53, which was categorized as moderate pollution. The average RI was 344.58 which represented a considerable ecological risk. PLI and RI shared a similar spatial distribution with high values centralized in the old industrial area in the southeast and railway stations for passengers and goods in the south of the city, followed by the old town within the ancient city wall, and low values located in the north and west areas. Cadmium was the main factor for both soil pollution and potential ecological risk primarily due to farmland topsoil in the eastern suburb of Kaifeng City with high Cd concentrations resulted from sewage irrigation deposited in the urban area by wind, human activities such as soot discharged from the chemical fertilizer plant of Kaifeng, transportation and coal combustion.
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Monitoramento Ambiental , Metais Pesados/análise , Poluentes do Solo/análise , Solo/química , Cádmio , China , Cidades , Medição de Risco , EsgotosRESUMO
OBJECTIVE: To investigate the expression of CD44 in leukemia cell lines and its role in adhesion, migration and infiltration of leukemia cells. METHODS: The expression levels of CD44 in four leukemia cell lines SHI-1, THP-1, NB4 and K562 were assayed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot when they were in logarithmic phase. And these cell lines were divided into control group (treated with same species and isotype IgG) and experimental group (treated with anti-CD44 mono-clonal antibody). The assays of cell-cell adhesion to endothelial cells line ECV304, migration through the artificial matrix membrane and infiltration through the Matrigel were performed. RESULTS: The relative expression ratios of CD44 to GAPDH in SHI-1, THP-1, NB4 cells were 0.0731 ± 0.0072, 0.0827 ± 0.0151 and 0.1473 ± 0.0365, respectively, which were significantly higher than that in K562 cells (0.0002 ± 0.0000, P < 0.01). Cell-cell adhesion assay showed that the adhesion rates of SHI-1, THP-1 and NB4 cells in the experimental group decreased to 72.78%, 64.09% and 57.42%, respectively, and were lower than those of the control groups, while that of K562 cells in the experimental group was 106.16%. Migration assay showed that the transmembrane rates of SHI-1,THP-1 and NB4 cells were 55%, 29% and 25% in the control group, respectively, and decreased to 32%, 18% and 12% in the experimental group, respectively, while those of K562 cells in both control group and experimental group remained 2%. The infiltration rates of SHI-1, THP-1 and NB4 cells decreased from 24%, 15% and 13% in the control group to 12%, 8% and 4% in the experimental group, respectively, while K562 cells in both groups could not pass through the Matrigel. CONCLUSION: CD44 antigen might play an important role in the adhesion, migration and infiltration of leukemia cells and be involved in the extra-medullary infiltration of leukemia cells.
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Receptores de Hialuronatos/metabolismo , Leucemia/metabolismo , Leucemia/patologia , Adesão Celular , Movimento Celular , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Células K562 , Invasividade NeoplásicaRESUMO
OBJECTIVE: To assess the efficacy and safety of local application of intraoperative mitomycin C (MMC) at the osteotomy site in primary external dacryocystorhinostomy (EX-DCR). METHODS: A comprehensive literature search of the Cochrane Library, PubMed and Embase was undertaken to identify relevant trials comparing EX-DCR with MMC (MMC group, from 0.2-1.0 mg/mL) and without MMC (control group). A total of nine randomized controlled trials (RCTs) were selected and a meta-analysis performed on the results of success rates, which were defined as patency of the nasolacrimal canal and symptomatic improvement. Statistical analysis was performed using RevMan 5.0 software. RESULTS: Nine RCTs reporting on a total of 562 DCRs including patients in the age range 30-57 years were included in the meta-analysis. However, the total number of males and females could not be determined as only four RCTs reported on this aspect. There was a significantly higher success rate in the MMC group in comparison with the control group (odds ratio, OR, 2.11; 95% confidence interval, CI, 1.19-3.74, P = 0. 01). In two RCTs, the mean osteotomy size 6 months postoperatively was significantly larger in the MMC group than in the control group (about 27mm(2) in the MMC group versus about 12mm(2) in the control group in the first study, and about 22mm(2) in the MMC group versus about 18mm(2) in the control group in the second study, P < 0.005). No intraoperative or postoperative complications except two cases with delayed healing of the external skin wound were recorded in the MMC group. CONCLUSION: Intraoperative MMC application seems to be a safe adjuvant that could reduce the closure rate of the osteotomy site after primary EX-DCR. Further well-organized, prospective, randomized studies involving larger patient numbers divided into subgroups for different concentrations of locally applied MMC are warranted.
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Alquilantes/administração & dosagem , Dacriocistorinostomia/métodos , Obstrução dos Ductos Lacrimais/tratamento farmacológico , Mitomicina/administração & dosagem , Adulto , Terapia Combinada , Bases de Dados Factuais , Feminino , Humanos , Cuidados Intraoperatórios , Masculino , Pessoa de Meia-Idade , Ducto Nasolacrimal/efeitos dos fármacos , Ducto Nasolacrimal/cirurgia , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do TratamentoRESUMO
Monodisperse Fe3O4 magnetic nanoparticles (NPs) were prepared under facile solvothermal conditions and successively functionalized with silica and Au to form core/shell Fe3O4@silica@Au NPs. Furthermore, the samples were used as matrix to construct a glucose sensor based on glucose oxidase (GOD). The immobilized GOD retained its bioactivity with high protein load of 3.92×10(-9) mol·cm(-2), and exhibited a surface-controlled quasi-reversible redox reaction, with a fast heterogeneous electron transfer rate of 7.98±0.6 s(-1). The glucose biosensor showed a broad linear range up to 3.97 mM with high sensitivity of 62.45 µA·mM(-1) cm(-2) and fast response (less than 5s).
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Compostos Férricos/química , Compostos Ferrosos/química , Glucose Oxidase/química , Glucose/química , Ouro/química , Nanopartículas de Magnetita/química , Oxigênio/química , Dióxido de Silício/química , Adsorção , Técnicas Biossensoriais/métodos , Catálise , OxirreduçãoRESUMO
Monodehydroascorbate reductase (MDAR; EC 1.6.5.4) is crucial for AsA regeneration and essential for maintaining the reduced pool of AsA. And the amino acid residue C117 of chloroplastic MDAR is the conserved cysteine residue in MDAR isoforms. A series mutation of conserved amino acid residue cysteine117 (C117) was constructed to investigate its role in MDAR structural stability and activity. Our study revealed that mutation in this conserved residue could cause pronounced loss of activity and conformational changes. Spectroscopic experiments indicated that these mutations influenced transition from the molten globule intermediate to the native state in folding process. These results suggested that amino acid residue C117 played a relatively important role in keeping MDAR structural stability and activity.