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This study used the brilliant cresyl blue (BCB) staining method to group buffalo oocytes (BCB+ and BCB-) and perform in vitro maturation, in vitro fertilization and embryo culture. At the same time, molecular biology techniques were used to detect gap junction protein expression and oxidative stress-related indicators to explore the molecular mechanism of BCB staining to predict oocyte developmental potential. The techniques of buffalo oocytes to analyse their developmental potential and used immunofluorescence staining to detect the expression level of CX43 protein, DCFH-DA probe staining to detect ROS levels and qPCR to detect the expression levels of the antioxidant-related genes SOD2 and GPX1. Our results showed that the in vitro maturation rate, embryo cleavage rate and blastocyst rate of buffalo oocytes in the BCB+ group were significantly higher than those in the BCB- group and the control group (p < .05). The expression level of CX43 protein in the BCB+ group was higher than that in the BCB- group both before and after maturation (p < .05). The intensity of ROS in the BCB+ group was significantly lower than that in the BCB- group (p < .05), and the expression levels of the antioxidant-related genes SOD2 and GPX1 in the BCB+ group were significantly higher than those in the BCB- group (p < .05). Brilliant cresyl blue staining could effectively predict the developmental potential of buffalo oocytes. The results of BCB staining were positively correlated with the expression of gap junction protein and antioxidant-related genes and negatively correlated with the reactive oxygen species level, suggesting that the mechanism of BCB staining in predicting the developmental potential of buffalo oocytes might be closely related to antioxidant activity.
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Búfalos , Conexina 43 , Técnicas de Maturação in Vitro de Oócitos , Oócitos , Oxazinas , Estresse Oxidativo , Animais , Oócitos/metabolismo , Conexina 43/genética , Conexina 43/metabolismo , Feminino , Técnicas de Maturação in Vitro de Oócitos/veterinária , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Superóxido Dismutase/genética , Glutationa Peroxidase/metabolismo , Glutationa Peroxidase/genética , Fertilização in vitro/veterinária , Técnicas de Cultura Embrionária/veterinária , Glutationa Peroxidase GPX1 , Desenvolvimento Embrionário/fisiologia , Coloração e Rotulagem , Antioxidantes/metabolismoRESUMO
Atresia is a process in ovarian follicles that is regulated by hormone-induced apoptosis. During atresia, granulosa cell (GC) apoptosis is a key mechanism orchestrated through diverse signaling pathways. Cocaine- and amphetamine-regulated transcript (CART) signaling within ovarian GCs has been demonstrated to play a key role in the regulation of follicular atresia in cattle, pigs, and sheep. The present work aimed to investigate the potential local regulatory role of CART in GC apoptosis-induced follicular atresia in buffalo, focusing on the modulation of the AKT/GSK3ß/ß-catenin signaling pathways, which are the intracellular signaling pathways involved in cell viability. Our findings revealed increased expression of CARTPT and BAX and decreased levels of AKT, ß-catenin, and CYP19A1 genes in atretic follicles compared to healthy follicles. Subsequently, CART treatment in the presence of FSH inhibited the FSH-induced increase in GC viability by reducing estradiol production and increasing apoptosis. This change was accompanied by an increase in the gene expression levels of both CARTPT and BAX. At the protein level, treatment with CART in the presence of FSH negatively affected the activity of AKT, ß-catenin, and LEF1, while the activity of GSK3ß was enhanced. In conclusion, our study shows how CART negatively influences buffalo GC viability, underlying the modulation of the AKT/GSK3ß/ß-catenin pathway and promoting apoptosis-a key factor in follicular atresia.
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Electrical impedance tomography (EIT) provides an indirect measure of the physiological state and growth of the maize ear by reconstructing the distribution of electrical impedance. However, the two-dimensional (2D) EIT within the electrode plane finds it challenging to comprehensively represent the spatial distribution of conductivity of the intact maize ear, including the husk, kernels, and cob. Therefore, an effective method for 3D conductivity reconstruction is necessary. In practical applications, fluctuations in the contact impedance of the maize ear occur, particularly with the increase in the number of grids and computational workload during the reconstruction of 3D spatial conductivity. These fluctuations may accentuate the ill-conditioning and nonlinearity of the EIT. To address these challenges, we introduce RFNetEIT, a novel computational framework specifically tailored for the absolute imaging of the three-dimensional electrical impedance of maize ear. This strategy transforms the reconstruction of 3D electrical conductivity into a regression process. Initially, a feature map is extracted from measured boundary voltage via a data reconstruction module, thereby enhancing the correlation among different dimensions. Subsequently, a nonlinear mapping model of the 3D spatial distribution of the boundary voltage and conductivity is established, utilizing the residual network. The performance of the proposed framework is assessed through numerical simulation experiments, acrylic model experiments, and maize ear experiments. Our experimental results indicate that our method yields superior reconstruction performance in terms of root-mean-square error (RMSE), correlation coefficient (CC), structural similarity index (SSIM), and inverse problem-solving time (IPST). Furthermore, the reconstruction experiments on maize ears demonstrate that the method can effectively reconstruct the 3D conductivity distribution.
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The conjugation of terminal ammonium salt groups with perovskite surfaces is a frequently employed technique that aims to enhance the overall performance of perovskite materials, encompassing both bulk and surface properties. Particularly, it exhibits heightened efficacy when applied to surface modification, due to its ability to mitigate defect accumulation and facilitate facile binding with the receptive sites inherent to the perovskite structure. However, the interaction of the bulk ammonium group with PbI2 has the potential to form a low-dimensional phase of perovskite, which may obstruct carrier extraction at the interface. Therefore, the surface passivators (MeO-PFACl) are designed through intramolecular potential manipulation. The combinations of the electron-donating methoxy group and π-π conjugation of the phenyl ring reduce the local potential at the reactive site of formamidinium group, making it less likely to form a low-dimension phase with perovskite. This surface passivation strategy effectively suppresses the surface nonradiative recombination and promotes the interface carrier extraction. The devices treated with MeO-PFACl have demonstrated exceptional performance, achieving a peak power conversion efficiency (PCE) of 25.88%, with an average PCE of 25.37%. These works offer a novel principle for enhancing both the efficiency and stability of PSCs using ammonium-incorporated molecules without the induction of an additional phase layer.
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The subcortical maternal complex, which consists of maternal-effect genes, plays a crucial role in the development of oocytes and preimplantation embryo until the activation of the zygote genome. One such gene, known as peptidyl-arginine deiminase VI (Padi6), is involved in the oocyte maturation, fertilization and embryonic development. However, the precise function of Padi6 gene in buffalo is still unclear and requires further investigation. In this study, the sequence, mRNA and protein expression patterns of Padi6 gene were analyzed in oocytes, preimplantation embryos and somatic tissues of buffalo. The coding sequence of gene was successfully cloned and characterized. Real-time quantitative PCR results indicated an absence of Padi6 transcripts in somatic tissues. Notably, the expression levels of Padi6 in oocytes showed an increased from the germinal vesicle stage to metaphase II stage, followed by a rapid decrease during the morula and blastocyst stages. Immunofluorescence analysis confirmed these findings, revealing a noticeable decline in protein expression levels. Our research provides the initial comprehensive expression profile of Padi6 in buffalo oocytes and preimplantation embryos, serving as a solid foundation for further investigations into the functionality of maternal-effect genes in buffalo.
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Surface properties of SnO2 and their effects on the growth of perovskite films play a crucial role for perovskite solar cells (PSCs). Herein, a facile strategy to synchronously regulate the buried interface defects and energy level arrangement, as well as improve the crystallinity of perovskite films with alleviated micro-strain by pre-modifying the SnO2 surface with ammonium hexafluorophosphate (NH4PF6) is proposed. The device achieved the promising PCE of 22.50% and improved stability.
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BACKGROUND: During mammalian pre-implantation embryonic development (PED), the process of maternal-to-zygote transition (MZT) is well orchestrated by epigenetic modification and gene sequential expression, and it is related to the embryonic genome activation (EGA). During MZT, the embryos are sensitive to the environment and easy to arrest at this stage in vitro. However, the timing and regulation mechanism of EGA in buffaloes remain obscure. RESULTS: Buffalo pre-implantation embryos were subjected to trace cell based RNA-seq and whole-genome bisulfite sequencing (WGBS) to draw landscapes of transcription and DNA-methylation. Four typical developmental steps were classified during buffalo PED. Buffalo major EGA was identified at the 16-cell stage by the comprehensive analysis of gene expression and DNA methylation dynamics. By weighted gene co-expression network analysis, stage-specific modules were identified during buffalo maternal-to-zygotic transition, and key signaling pathways and biological process events were further revealed. Programmed and continuous activation of these pathways was necessary for success of buffalo EGA. In addition, the hub gene, CDK1, was identified to play a critical role in buffalo EGA. CONCLUSIONS: Our study provides a landscape of transcription and DNA methylation in buffalo PED and reveals deeply the molecular mechanism of the buffalo EGA and genetic programming during buffalo MZT. It will lay a foundation for improving the in vitro development of buffalo embryos.
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Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) modulates T helper cell differentiation and nuclear factor-κB (NF-κB) pathway-mediated inflammation and potentially regulates lipid metabolism, which are all critical factors involved in atherosclerosis. The present study aimed to investigate the effect of MALT1 on the cellular functions of proatherogenic vascular smooth muscle cells (VSMCs). Therefore, to establish a human proatherogenic VSMC model, VSMCs were treated with different doses of oxidized low-density lipoprotein (oxLDL). Subsequently, the effect of MALT1 overexpression or knockdown in proatherogenic VSMCs treated with or without NF-κB activator was also explored. The results showed that treatment of proatherogenic VSMCs with oxLDL significantly elevated the mRNA and protein expression levels of MALT1 in a dose-dependent manner. Furthermore, MALT1 overexpression enhanced cell viability, invasion and phenotype switching and reduced apoptosis in proatherogenic VSMCs. However, MALT1 knockdown exerted the opposite effect on the above cellular functions. Additionally, the results revealed that MALT1 could positively regulate the NF-κB pathway in proatherogenic VSMCs. Moreover, treatment of proatherogenic VSMCs with NF-κB activator not only exacerbated the dysregulation of cellular functions, but also hampered the effect of MALT1 knockdown on attenuating cell growth, invasion and synthetic phenotype switching, thus suggesting that NF-κB was essential for the regulation of MALT1-triggered functions in proatherogenic VSMCs. In conclusion, the current study suggested that MALT1 could exacerbate cell viability, mobility and synthetic phenotype switching of proatherogenic VSMCs in a NF-κB signaling-dependent manner. Therefore, MALT1 could be considered as a potential therapeutic target for atherosclerosis.
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Apelin (APLN) was believed to be an adipokine secreted from adipose tissue. However, studies demonstrate that it is a pleiotropic peptide and has several effects on the female reproductive system. In this study, We examined the effects of different doses of IGF1 and FSH in the presence of APLN-13 on the production of progesterone in buffalo ovary granulosa cells. Furthermore, different doses of APLN isoforms (APLN-13 and APLN-17) were tested on proliferation, Bax protein expression, and antioxidant capacity in the same cells. Granulosa cells of buffalo ovaries were cultured in the presence of different doses of IGF1 and FSH with or without APLN-13 (10-9 M) to evaluate its effect on the secretion of progesterone tested by ELISA assay. The WST-1 method was used to survey the effect of APLN on granulosa cell proliferation and cytotoxicity. In addition, the antioxidant capacity of the cells in the presence of APLN was assessed using the FRAP method. mRNA and Bax protein levels were measured in granulosa cells treated with APLN using real-time PCR and western blot techniques. APLN-13 (10-9) stimulated the effect of IGF1 on the production of progesterone, and its levels were affected by APLN-13 dose-dependently. However, it did not significantly stimulate the effect of FSH on the secretion of progesterone. APLN-13 (all doses) and APLN-17 (10-8 and 10-9 M) improved the proliferation of granulosa cells. Moreover, preincubation of the cells for an hour by APLN receptor antagonist (ML221, 10 µM) did not significantly affect the proliferation of cells induced by APLN. Neither APLN-13 nor APLN-17 were not cytotoxic for the cells compared to the control treatment. APLN-13 at the doses of 10-6 and 10-8 M substantially up and down-regulated Bax protein expression; however, such effects were not observed when the cells were preincubated with ML221. In addition, APLN-17 did not influence the expression amount of Bax. Furthermore, both APLN-13 and -17 improved the total antioxidant capacity of the ovarian granulosa cells, but such effects were not seen when the cells were preincubated with ML221. According to these results, APLN enhanced the steroidogenesis induced by IGF1 but did not affect the steroidogenesis induced by FSH. APLN also enhanced the cell proliferation and antioxidant capacity of buffalo ovaries follicular granulosa cells; however, its effect on Bax expression was different.
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Búfalos , Progesterona , Feminino , Animais , Antioxidantes/farmacologia , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo , Proteína X Associada a bcl-2/farmacologia , Apelina/genética , Apelina/metabolismo , Apelina/farmacologia , Folículo Ovariano/metabolismo , Células da Granulosa/metabolismo , Proliferação de Células , Hormônio Foliculoestimulante/farmacologia , Hormônio Foliculoestimulante/metabolismo , Células CultivadasRESUMO
Defect accumulation and nonradiative recombination at the interface of the electron-transport layer (ETL) and the photosensitive layer are inevitable obstacles to efficient and stable perovskite solar cells (PSCs). Herein, we reported a dual-effect interface modification strategy that employs potassium tetrafluoroborate (KBF4) molecules for the simultaneous passivation of the SnO2/perovskite interface and perovskite grain boundaries. The introduced highly electronegative BF4- enriched at the SnO2 surface and the chemical bond interaction between them can effectively reduce the hydroxyl (-OH) group defects on the surface of SnO2, improve electron mobility, and reduce nonradiative recombination. Meanwhile, partial K+ diffuses into the grain boundaries, causing the halogen ions to be uniformly distributed in the perovskite film and resulting in better crystallinity. Therefore, the performance of the experimental device was improved from 20.34 to 22.90% compared with the reference device, with a high electrical performance (JSC = 25.1 mA cm-2, VOC = 1.137 V). In particular, the unencapsulated target PSCs retained 85% of their original PCE after aging for 1000 h under ambient conditions (70 ± 10% RH) in the dark.
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Aneuploidy is one of the main causes of fetal and embryonic mortality in mammals. Nonetheless, its incidence in domestic ruminants has been investigated little. Indeed, no incidence data have ever been reported for water buffalo. To establish the incidence of aneuploidy in this species, we analysed in vitro matured metaphase II (MII) oocytes with corresponding first polar bodies (I PB) of the river (2n = 50) and swamp (2n = 48) buffaloes. For the first time, six river type probes (corresponding to chromosomes 1-5 and heterosome X), were tested on swamp buffalo metaphases using Multicolor-Fluorescent In Situ Hybridization (M-FISH) before their use on oocytes MII metaphases. Of the 120 total Cumulus Oocyte Complexes (COCs, 60 for each buffalo type) subjected to in vitro maturation, 104 reached the MII stage and were analysed by M-FISH. Haploid chromosome arrangement and visible I PB were observed in 89 of the oocytes (45 in river and 44 in swamp type). In the river type, the analysis revealed one oocyte was disomic for the chromosome X (2.22%). In the swamp type, one oocyte was found to be nullisomic for chromosome X (2.27%); another was found to be nullisomic for chromosome 5 (2.27%). We also observed one oocyte affected by a premature separation of sister chromatids (PSSC) on the chromosome X (2.27%). In both buffalo types, no abnormalities were detected in other investigated chromosomes. Based on merged data, the overall aneuploidy rate for the species was 3.37%. Oocytes with unreduced chromosomes averaged 1.92% across the two types, with 1.96% in river and 1.88% in swamp. The interspecies comparison between these data and cattle and pig published data revealed substantial difference in both total aneuploidy and diploidy rates. Reducing the negative impact of the meiotic segregation errors on the fertility is key to more sustainable breeding, an efficient embryo transfer industry and ex-situ bio-conservation. In this respect, additional M-FISH studies are needed on oocytes of domestic species using larger sets of probes and/or applying next generation sequencing technologies.
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Bison , Búfalos , Aneuploidia , Animais , Búfalos/genética , Bovinos , Hibridização in Situ Fluorescente , Oócitos , Rios , Suínos , Cromossomo XRESUMO
All-inorganic CsPbI3 perovskite solar cells (PSCs) are becoming desirable for their excellent photovoltaic ability and adjustable crystal structure distortion. However, the unsatisfactory crystallization of the perovskite phase is unavoidable and leads to challenges on the road to the development of high-quality CsPbI3 perovskite films. Here, we reported the intermediate-phase-modified crystallization (IPMC) method, which introduces pyrrolidine hydroiodide (PI) before the formation of the perovskite phase. The hydrogen bonding, which originates from the interaction between the -NH in PI and the dimethylammonium iodide (DMAI) from the precursor solution, improved the crystallization conditions and further prompted the transition from the DMAPbI3 phase to CsPbI3 perovskite phase. The application of the IPMC method not only decreased the trap density but also changed the energy alignment for better separation of electron-hole pairs. As a result, the devices based on the PI-CsPbI3 perovskite films reached an efficiency of 18.72% and maintained 85% of their initial PCE after 1000 h of being stored in an ambient environment (â¼25% RH, 25 °C). This work stimulates inspiration on how to conveniently fabricate high-quality perovskite films in industry.
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Apelin (APLN), as a ligand for APJ (an orphan G-protein-coupled receptor), is an adipokine with pleiotropic effects in many physiological processes of the body. It has an important role in the control of reproduction particularly in females (mainly in control of ovarian function). This study was carried out to investigate the mRNA and protein amounts of APLN/APJ in granulose cells (GCs) of ovarian follicles with small (SF), medium (MF), and large (LF) sizes of buffalo (Bubalus bubalis) and the effect of IGF1 and follicle-stimulating hormone (FSH) on the expression levels of APLN/APJ. In addition, we evaluated the effect of various doses of APLN (isoforms -13 and -17) singly or in combination with IGF1 and FSH on estradiol (E2) and progesterone (P4) secretion in GCs. The mRNA and protein abundance of APLN was the highest in GCs of LF while the APJ expression enhanced with follicle enlargement in GCs (p-value <0.01). IGF1 and FSH elevated the mRNA and protein amounts of APLN and FSH, and IGF1 increased the expression of APJ in buffalo GCs (p-value <0.01). Both isoforms of APLN (-13/-17) singly or in the presence of IGF1 or FSH increased the secretion of E2 and P4 with or without preincubation of cells with APJ antagonist (ML221 10 µM), although we had some variation in the effects. Concurrently, APLN-13/-17 significantly increased the mRNA and protein expression of CYP19A1 and StAR (p-value <0.01). ML221 substantially diminished the secretion of E2 and P4 and also the expression of CY19A1 and StAR in buffalo GCs (p-value <0.01). We also revealed that APLN-13/-17 (10-9 M), singly or in response to IGF1 and FSH, increased the production of E2 and P4 in different times of stimulation. In conclusion, APLN may play a crucial role in steroidogenesis and follicular development in ovarian GCs of buffalo.
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Búfalos , Ovário , Animais , Apelina/genética , Apelina/metabolismo , Apelina/farmacologia , Receptores de Apelina/metabolismo , Feminino , Células da GranulosaRESUMO
Two-dimensional/three-dimensional (2D/3D) Ruddlesden-Popper perovskite materials have shown the enormous potential to achieve both efficient and stable photovoltaic devices for commercial applications. Unfortunately, the single function of spacer cations limits their further improvements in efficiency to reach values as high as those of 3D perovskites. Herein, we developed a new-type multifunctional heterocyclic-based spacer cation of 2-(methylthio)-4,5-dihydro-1H-imidazole (MTIm+) to achieve a synchronous improvement of efficiency and stability for 2D/3D perovskite solar cells (PSCs). Owing to the presence of special chemical groups (imidazole and methylthio), strong interactions have been found between MTIm+ and the 3D perovskite component, leading to an excellent passivation effect. More important, at the initial stage of crystallization, uniform nucleation distribution would be generated around the spacer cation, which is helpful for improved crystallinity and reduced growth defects. The smaller layer space compared to that of cations based on aromatic hydrocarbons caused effective carrier transfer between inorganic layers in 2D/3D perovskites. As a result, the 2D/3D (n = 30) PSCs based on MTIm exhibit a champion PCE up to 21.25% with a high Voc of 1.14 V. Besides, the 2D/3D perovskite devices have realized dramatically enhanced humidity and thermal stability, maintaining 94% of the starting PCE enduring aging at about 50% RH for 2880 h and at 85 °C for 360 h, respectively. We believe that it would provide a significant strategy to further promote the photovoltaic performances and the long-term stability of 2D/3D perovskite devices toward future practical applications.
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Curcumin (Cur) is a flavonoid derived from Curcuma longa L. that has been shown to have a variety of biological activities, but some previous studies have described its non-negligible negative effects on female reproduction and embryo development. To further explore the toxic stress effect, this study investigated apoptosis and autophagy of healthy buffalo (Bubalus bubalis) derived granulosa cells (GCs) exposed to Cur and/or autophagy inhibitors. Results showed that Cur declined viability of GCs in a concentration-dependent manner. Apoptosis was observed in Cur-treated GCs from 3 h. Meanwhile, under Cur stress, autophagosomes accumulated in cells, and the expression levels of autophagy key proteins LC3 and Beclin 1 were up-regulated, suggesting that Cur could induce autophagy in GCs. Early autophagy inhibitor 3-methyladenine (3-MA) increased the apoptosis rate of Cur exposed GCs, but the autophagosome degradation inhibitor chloroquine (CQ) had no effect on the apoptosis rate. The network pharmacological and molecular docking analysis indicated that the perturbation of IKK/NF-κB might be the cause of Cur toxicity toward GCs. This study unveiled another side of Cur pharmacological effects that programmed cell death can be induced by Cur in GCs, suggesting that it should be prudent to use Cur as a clinical drug for its side effects on the female reproductive system.
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Curcumina , Feminino , Animais , Curcumina/toxicidade , Simulação de Acoplamento Molecular , Proteína Beclina-1/farmacologia , NF-kappa B , Farmacologia em Rede , Autofagia , Apoptose , Células da Granulosa/metabolismo , Flavonoides/farmacologia , Cloroquina/toxicidadeRESUMO
OBJECTIVES: Long non-coding RNAs (lncRNAs) play a crucial part in cancer progression. However, in epithelial ovarian carcinoma (EOC), the role of SNHG22 needs to be further explained. METHODS: Quantitative real-time PCR was used to detect the expression of SNHG22. EOC cells were stably transfected with lentivirus approach and cell proliferation, glycolysis and cell apoptosis, as well as tumorigenesis in animal were performed to assess the effects of SNHG22 in EOC. Chromatin immunoprecipitation (ChIP) and luciferase reporter assay were conducted to confirm the relationship between SP1 and SNHG22. RESULTS: Higher expressed SNHG22 was associated with a poor prognosis in EOC tissues. SNHG22 facilitated glycolysis and proliferation. Mechanistically, LDHA deficiency and glycolysis inhibitor (2-DG, 3-BG) partly rescued proliferation. SP1 mediated SNHG22 expression at the transcriptional level and the SNHG22 promoter region (-900~ -600) was necessary for SP1 binding. Hypoxia and HIF-1α also upregulated SNHG22 expression. CONCLUSION: SNHG22 is an independent prognostic biomarker for EOC. SNHG22 promotes EOC progression and is a prospective therapeutic target.
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Although the power conversion efficiencies (PCEs) of perovskite solar cells (PSCs) have made great progress, the surface and interface defects still affect their PCE and stability and hinder the commercialization. To overcome this problem, 1-methylimidazole (1-MIm) and 1-methylbenzimidazole (1-MBIm) were used as the interfacial passivation agents to passivate the defects at surface and interface. The results indicated that, in contrast to 1-MIm, 1-MBIm displayed a stronger Lewis coordination interaction with the uncoordinated Pb2+ to reduce the non-radiative recombination and also effectively improved the charge transfer capacity of perovskite films due to its strong π-π conjugate interaction, resulting in the better photovoltaic performance. As a result, the PCE of the champion 1-MBIm PSC was improved from 19.48 (pristine) to 21.22 % with a dramatically enhanced open-circuit voltage (Voc =1.15â V). More importantly, a significant improvement in long-term stability was achieved for 1-MBIm perovskite devices, which was attributed to the high-quality perovskite film caused by the strong passivation effect of 1-MBIm and the hydrogen bond with water molecules. The results offers an efficient and facile strategy by interface engineering to fabricate high-performance and stable PSCs for commercial application.
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This study was performed to investigate the difference in developmental competence of oocytes derived from ovum pick-up (OPU) and slaughterhouse ovaries (SLH), and its underlying mechanisms. The OPU and SLH oocytes were in-vitro maturated and fertilized to produce blastocysts, and these blastoycsts were collected to explore the expression of key genes for developmental potential and telomere (Oct-4, Sox2, Nanog, Cdx2, Gata3, E-cadherin, ß-catenin, TERT, TERF1 and TERF2). The results showed that both the cleavage and blastocyst rates were significantly higher for the OPU group (68.31%, 39.48%, respectively) than SLH group (57.59%, 26.50%, respectively) (P < 0.01). The relative mRNA abundances of Sox2, Oct-4, Nanog and E-cadherin were significantly higher in the OPU blastocysts than the SLH ones (P < 0.01). Protein expression analysis by Western blot and immunofluorescence also revealed that the expression of E-cadherin and Sox2 was significantly higher in OPU blastocysts than SLH ones. However, there was no significant differences between the two groups in the expression of Cdx2, ß-catenin, Gata3, TERT, TERF1, TERF2. These results imply oocyte sources modify the expression of development and adhesion related genes in blastocysts, which may elucidate a possible reasoning for the low development competence of buffalo SLH embryos.
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Matadouros , Búfalos , Animais , Blastocisto , Búfalos/genética , Fertilização in vitro/veterinária , Oócitos , ÓvuloRESUMO
This work aimed to investigate the effect of fucoidan (FPS) on urate transporters induced by uric acid (UA). The results showed that UA stimulated the expression of glucose transporter 9 (GLUT9) and urate transporter 1 (URAT1) in HK-2 cells, and FPS could reverse the effect. Moreover, UA could activate NF-κB, JNK and PI3K/Akt pathways, but both pathway inhibitors and FPS inhibited the UA-induced activation of these three pathways. These data suggested that FPS effectively inhibited the expression induction of reabsorption transporters URAT1 and GLUT9 by UA, through repressing the activation of NF-κB, JNK and PI3K/Akt signal pathways in HK-2 cells. The in vitro research findings support the in vivo results that FPS reduces serum uric acid content in hyperuricemia mice and rats through inhibiting the expression of URAT1 and GLUT9 in renal tubular epithelial cells. This study provides a theoretical basis for the application of FPS in the treatment of hyperuricemia.
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Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Supressores da Gota/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Túbulos Renais Proximais/efeitos dos fármacos , Laminaria , NF-kappa B/metabolismo , Transportadores de Ânions Orgânicos/metabolismo , Proteínas de Transporte de Cátions Orgânicos/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Polissacarídeos/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Linhagem Celular , Supressores da Gota/isolamento & purificação , Humanos , Túbulos Renais Proximais/enzimologia , Laminaria/química , Polissacarídeos/isolamento & purificação , Transdução de Sinais , Ácido Úrico/toxicidadeRESUMO
OBJECTIVE: We aimed to investigate the expression of long noncoding RNA- (lncRNA-) HEIH in patients with coronary artery disease (CAD) and its impact on patients' prognosis. Patients and Methods. From July 2015 to December 2018, 250 patients who underwent coronary angiography, including 50 in the control group and 150 in the CAD group, were collected for detection of the expression of lncRNA-HEIH by real-time quantitative polymerase chain reaction (qPCR). The severity of CAD was evaluated through SYNTAX scoring system. In addition, these patients with CAD were followed up for 3 years, and the major cardiac adverse events such as myocardial infarction and revascularization were recorded. RESULTS: The expression of lncRNA-HEIH in plasma of patients with CAD was remarkably higher than that in the control subjects and was verified to be relevant to the severity of CAD. Meanwhile, it was found that CAD patients with high expression of lncRNA-HEIH had higher rates of dyslipidemia as well as CAD family history and higher overall incidence of major cardiac adverse events than those with low expression of lncRNA-HEIH. CONCLUSIONS: lncRNA-HEIH expression is upregulated in the plasma of CAD patients, which is capable of affecting the prognosis of patients.