Arabidopsis mutants produced by constitutive overexpression of the CRISPR/Cas9 genome editing system are usually mosaics in the T1 generation. In this study, we used egg cell-specific promoters to ...drive the expression of Cas9 and obtained non-mosaic T1 mutants for multiple target genes with high efficiency. Comparisons of 12 combinations of eight promoters and two terminators found that the efficiency of the egg cell-specific promoter-controlled CRISPR/Cas9 system depended on the presence of a suitable terminator, and the composite promoter generated by fusing two egg cell-specific promoters resulted in much higher efficiency of mutation in the T1 generation compared with the single promoters.
To accelerate the application of the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/ CRISPR-associated protein 9) system to a variety of plant species, a toolkit with ...additional plant selectable markers, more gRNA modules, and easier methods for the assembly of one or more gRNA expression cassettes is required.
We developed a CRISPR/Cas9 binary vector set based on the pGreen or pCAMBIA backbone, as well as a gRNA (guide RNA) module vector set, as a toolkit for multiplex genome editing in plants. This toolkit requires no restriction enzymes besides BsaI to generate final constructs harboring maize-codon optimized Cas9 and one or more gRNAs with high efficiency in as little as one cloning step. The toolkit was validated using maize protoplasts, transgenic maize lines, and transgenic Arabidopsis lines and was shown to exhibit high efficiency and specificity. More importantly, using this toolkit, targeted mutations of three Arabidopsis genes were detected in transgenic seedlings of the T1 generation. Moreover, the multiple-gene mutations could be inherited by the next generation.
We developed a toolkit that facilitates transient or stable expression of the CRISPR/Cas9 system in a variety of plant species, which will facilitate plant research, as it enables high efficiency generation of mutants bearing multiple gene mutations.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Liver sinusoidal endothelial cells (LSECs) critically regulate liver homeostasis and diseases through angiocrine factors. Notch is critical in endothelial cells (ECs). In the current study, Notch ...signaling was activated by inducible EC‐specific expression of the Notch intracellular domain (NIC). We found that endothelial Notch activation damaged liver homeostasis. Notch activation resulted in decreased fenestration and increased basement membrane, and a gene expression profile with decreased LSEC‐associated genes and increased continuous EC‐associated genes, suggesting LSEC dedifferentiation. Consistently, endothelial Notch activation enhanced hepatic fibrosis (HF) induced by CCl4. Notch activation attenuated endothelial nitric oxide synthase (eNOS)/soluble guanylate cyclase (sGC) signaling, and activation of sGC by 3‐(5′‐hydroxymethyl‐2′‐furyl)‐1‐benzylindazole (YC‐1) reversed the dedifferentiation phenotype. In addition, Notch activation subverted the hepatocyte‐supporting angiocrine profile of LSECs by down‐regulating critical hepatocyte mitogens, including Wnt2a, Wnt9b, and hepatocyte growth factor (HGF). This led to compromised hepatocyte proliferation under both quiescent and regenerating conditions. Whereas expression of Wnt2a and Wnt9b was dependent on eNOS‐sGC signaling, HGF expression was not rescued by the sGC activator, suggesting heterogeneous mechanisms of LSECs to maintain hepatocyte homeostasis. Conclusion: Endothelial Notch activation results in LSEC dedifferentiation and accelerated liver fibrogenesis through eNOS‐sGC signaling, and alters the angiocrine profile of LSECs to compromise hepatocyte proliferation and liver regeneration (LR). (Hepatology 2018).
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Salvia miltiorrhiza Bunge (SM) is a very popular medicinal plant that has been extensively applied for many years to treat various diseases, especially coronary heart diseases and cerebrovascular ...diseases, either alone or in combination with other Chinese plant-based medicines. Although a large number of studies on SM have been performed, they are scattered across a variety of publications. The present review is an up-to-date summary of the published scientific information about the traditional uses, chemical constituents, pharmacological effects, side effects, and drug interactions with SM, in order to lay the foundation for further investigations and better utilization of SM. SM contains diverse chemical components including diterpenoid quinones, hydrophilic phenolic acids, and essential oils. Many pharmacological studies have been done on SM during the last 30 years, focusing on the cardiovascular and cerebrovascular effects, and the antioxidative, neuroprotective, antifibrotic, anti-inflammatory, and antineoplastic activities. The research results strongly support the notion that SM has beneficial therapeutic properties and has a potential of being an effective adaptogenic remedy.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
Background and Purpose
Icariin, a major active ingredient in traditional Chinese medicines, is attracting increasing attention because of its unique pharmacological effects against ischaemic heart ...disease. The histone deacetylase, sirtuin‐1, plays a protective role in ischaemia/reperfusion (I/R) injury, and this study was designed to investigate the protective role of icariin in models of cardiac I/R injury and to elucidate the potential involvement of sirtuin‐1.
Experimental Approach
I/R injury was simulated in vivo (mouse hearts), ex vivo (isolated rat hearts) and in vitro (neonatal rat cardiomyocytes and H9c2 cells). Prior to I/R injury, animals or cells were exposed to icariin, with or without inhibitors of sirtuin‐1 (sirtinol and SIRT1 siRNA).
Key Results
In vivo and in vitro, icariin given before I/R significantly improved post‐I/R heart contraction and limited the infarct size and leakage of creatine kinase‐MB and LDH from the damaged myocardium. Icariin also attenuated I/R‐induced mitochondrial oxidative damage, decreasing malondialdehyde content and increasing superoxide dismutase activity and expression of Mn‐superoxide dismutase. Icariin significantly improved mitochondrial membrane homeostasis by increasing mitochondrial membrane potential and cytochrome C stabilization, which further inhibited cell apoptosis. Sirtuin‐1 was significantly up‐regulated in hearts treated with icariin, whereas Ac‐FOXO1 was simultaneously down‐regulated. Importantly, sirtinol and SIRT1 siRNA either blocked icariin‐induced cardioprotection or disrupted icariin‐mediated mitochondrial homeostasis.
Conclusions and Implications
Pretreatment with icariin protected cardiomyocytes from I/R‐induced oxidative stress through activation of sirtuin‐1 /FOXO1 signalling.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
By adopting the differential age method, we select 17 832 luminous red galaxies from the Sloan Digital Sky Survey Data Release Seven covering redshift 0 < z < 0.4 to measure the Hubble parameter. ...Using the full spectrum fitting package UlySS, these spectra are reduced with single stellar population models and optimal age information our selected sample is derived. With the decreasing age-redshift relation, four new observational H(z) data (OHD) points are obtained, which are H(z) = 69.0 + or - 19.6 km s super(-1) Mpc super(-1) at z = 0.07, H(z) = 68.6 + or - 26.2 km s super(-1) Mpc super(-1) at z = 0.12, H(z)=72.9 + or - 29.6 km s super(-1) Mpc super(-1) at z = 0.2 and H(z)=88.8 + or - 36.6 km s super(-1) Mpc super(-1) at z = 0.28, respectively. Combined with 21 other available OHD data points, the performance of the constraint on both flat and non-flat LambdaCDM models is presented.
Near infrared light photodiodes have been attracting increasing research interest due to their wide application in various fields. In this study, the fabrication of a new n‐type GaAs nanocone ...(GaAsNCs) array/monolayer graphene (MLG) Schottky junction is reported for NIR light detection. The NIR photodetector (NIRPD) shows obvious rectifying behavior with a turn‐on voltage of 0.6 V. Further device analysis reveals that the photovoltaic NIRPDs are highly sensitive to 850 nm light illumination, with a fast response speed and good spectral selectivity at zero bias voltage. It is also revealed that the NIRPD is capable of monitoring high‐switching frequency optical signals (∼2000 Hz) with a high relative balance. Theoretical simulations based on finite difference time domain (FDTD) analysis finds that the high device performance is partially associated with the optical property, which can trap most incident photons in an efficient way. It is expected that such a self‐driven NIRPD will have potential application in future optoelectronic devices.
A new Schottky junction near‐infrared light photodetector is fabricated by coating a GaAs nanocone array with a monolayer graphene film, which shows high sensitivity to near‐infrared light irradiation, with good reproducibility, excellent selectivity, and rapid response speed.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Background
Diabetic retinopathy (DR) is a common complication of diabetes mellitus that poses a threat to adults. MicroRNAs (miRNAs) play a key role in DR progression. However, the role and mechanism ...of miR‐192‐5p in DR remain unclear. We aimed to investigate the effect of miR‐192‐5p on cell proliferation, migration and angiogenesis in DR.
Methods
Expression of miR‐192‐5p, ELAV‐like RNA binding protein 1 (ELAVL1) and phosphoinositide 3‐kinase delta (PI3Kδ) in human retinal fibrovascular membrane (FVM) samples and human retinal microvascular endothelial cells (HRMECs) was assessed using RT‐qPCR. ELAVL1 and PI3Kδ protein levels were evaluated by Western blot. RIP and dual luciferase reporter assays were performed to confirm the miR‐192‐5p/ELAVL1/PI3Kδ regulatory networks. Cell proliferation, migration and angiogenesis were assessed by CCK8, transwell and tube formation assays.
Results
MiR‐192‐5p was decreased in FVM samples from DR patients and high glucose (HG)‐treated HRMECs. Functionally, overexpressed miR‐192‐5p inhibited cell proliferation, migration and angiogenesis in HG‐treated HRMECs. Mechanically, miR‐192‐5p directly targeted ELAVL1 and decreased its expression. We further verified that ELAVL1 bound to PI3Kδ and maintained PI3Kδ mRNA stability. Rescue analysis demonstrated that the suppressive effects of HG‐treated HRMECs caused by miR‐192‐5p up‐regulation were overturned by overexpressed ELAVL1 or PI3Kδ.
Conclusion
MiR‐192‐5p attenuates DR progression by targeting ELAVL1 and reducing PI3Kδ expression, suggesting a biomarker for the treatment of DR.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
The mammalian target of rapamycin (mTOR) pathway plays critical roles in intrinsic chemoresistance by regulating Fanconi anaemia complementation group D2 (FANCD2) expression. However, the mechanisms ...by which mTOR regulates FANCD2 expression and related inhibitors are not clearly elucidated. Extracts of Centipeda minima (C. minima) showed promising chemosensitizing effects by inhibiting FANCD2 activity. Here, we have aimed to identify the bioactive chemosensitizer in C. minima extracts and elucidate its underlying mechanism.
The chemosensitizing effects of arnicolide C (ArC), a bioactive compound in C. minima, on non-small cell lung cancer (NSCLC) were investigated using immunoblotting, immunofluorescence, flow cytometry, the comet assay, small interfering RNA (siRNA) transfection and animal models. The online SynergyFinder software was used to determine the synergistic effects of ArC and chemotherapeutic drugs on NSCLC cells.
ArC had synergistic cytotoxic effects with DNA cross-linking drugs such as cisplatin and mitomycin C in NSCLC cells. ArC treatment markedly decreased FANCD2 expression in NSCLC cells, thus attenuating cisplatin-induced FANCD2 nuclear foci formation, leading to DNA damage and apoptosis. ArC inhibited the mTOR pathway and attenuated mTOR-mediated expression of E2F1, a critical transcription factor of FANCD2. Co-administration of ArC and cisplatin exerted synergistic anticancer effects in the A549 xenograft mouse model by suppressing mTOR/FANCD2 signalling in tumour tissues.
ArC suppressed DNA cross-linking drug-induced DNA damage response by inhibiting the mTOR/E2F1/FANCD2 signalling axis, serving as a chemosensitizing agent. This provides insight into the anticancer mechanisms of ArC and offers a potential combinatorial anticancer therapeutic strategy.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
We report on the simple fabrication of monolayer graphene (MLG)/germanium (Ge) heterojunction for infrared (IR) light sensing. It is found that the as-fabricated Schottky junction detector exhibits ...obvious photovoltaic characteristics, and is sensitive to IR light with high I light/I dark ratio of 2 × 104 at zero bias voltage. The responsivity and detectivity are as high as 51.8 mA W–1 and 1.38 × 1010 cm Hz1/2 W–1, respectively. Further photoresponse study reveals that the photovoltaic IR detector displays excellent spectral selectivity with peak sensitivity at 1400 nm, and a fast light response speed of microsecond rise/fall time with good reproducibility and long-term stability. The generality of the above results suggests that the present MLG/Ge IR photodetector would have great potential for future optoelectronic device applications.
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IJS, KILJ, NUK, PNG, UL, UM
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