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Coronavirus disease 2019 (COVID-19) is an infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Since the start of the pandemic, olfactory dysfunction (OD) has been reported as a common symptom of COVID-19. In some asymptomatic carriers, OD is often the first and even the only symptom. At the same time, persistent OD is also a long-term sequela seen after COVID-19 that can have a serious impact on the quality of life of patients. However, the pathogenesis of post-COVID-19 OD is still unclear, and there is no specific treatment for its patients. The aim of this paper was to review the research on OD caused by SARS-CoV-2 infection and to summarize the mechanism of action, the pathogenesis, and current treatments. [ABSTRACT FROM AUTHOR]

The olfactory nerve (ON) is the only cranial nerve exposed to the external environment. Hence, it is susceptible to damage from head trauma, viral infection, inflammatory stimulation, and chemical toxins, which can lead to olfactory dysfunction. However, compared with all other cranial nerves, the ON is unique due to its inherent ability to regenerate. This characteristic provides a theoretical basis for treatment of olfactory dysfunction. Olfactory training (OT) is one of the main treatments for olfactory dysfunction. It is easy to apply and has few side-effects, and has been shown to be efficacious for patients with olfactory dysfunction of various causes. To further understand the application value of ON regeneration and OT on olfactory dysfunction, we review the research progress on the mechanism of ON regeneration and OT. [ABSTRACT FROM AUTHOR]

A new sum frequency generation imaging microscope using a novel sampling theory, compressive sensing (CS), has been developed for surface studies. CS differentiates itself from the conventional sampling methods by collecting fewer measurements than the traditional methods to reconstruct a high quality image. Pseudorandom patterns were applied to a light modulator and reflected the sum frequency (SF) signal generated from the sample into a photomultiplier tube detector. The image of the sample was reconstructed using sparsity preserving algorithms from the SF signal. The influences of the number of CS testing patterns applied and the number of SF pulses acquired for each pattern on the quality of the images was investigated and a comparison of the image quality with the traditional raster scan was made at varying resolutions for a gold patterned Si surface. Our results demonstrate the CS technique achieved 16 times the pixel density beyond the resolution where the raster scan strategy lost its ability to image the sample due to the dilution of the SF signal below the detection limit of the detector. [ABSTRACT FROM AUTHOR]

CRISPR/Cas9-mediated programmed cell death protein 1 (PD-1) disruption in chimeric antigen receptor (CAR) T cells could be an appealing choice to improve the therapeutic efficacy of CAR T cells in an immunosuppressive tumor microenvironment. In most of the reported cases, Cas9 was delivered into T cells by way of electroporation with RNA or protein. However, transient expression of Cas9 by transfection with a plasmid encoding its gene is apparently simpler, as it avoids the steps of in vitro transcription of DNA or protein production. This study tried nucleofection into human primary T cells of plasmids encoding both CRISPR/Cas9 for disrupting the PD-1 gene and the piggyBac transposon system for expressing CD133-specific CAR in one reaction. Based on drug selection, CD133-specific CAR T cells were obtained in which, on average, 91.5% of the PD-1 gene sites were disrupted, but almost no Cas9 gene expression was found in the final engineered CAR T cells. The PD-1-deficient CD133-specific CAR T cells showed similar levels of cytokine secretion and improved proliferation and cytotoxicity in vitro, and enhanced inhibition of tumor growth in an orthotopic mouse model of glioma, compared to conventional CD133-CAR T cells. The described method could be useful for the production of PD-1-deficient CAR T cells for cancer immunotherapy. [ABSTRACT FROM AUTHOR]

Based on the analysis of band structure and edge states on zigzag graphene nanoribbons (ZGNRs), we can study theoretically the origination of two minimal quantum conductance. At the two energy points − 0.20 eV and 0.15 eV corresponding to the two dips of quantum conductance, the spin-polarized quantum conductance is about 45%. Furthermore, the two types of edge-localized carriers in the opposite transport directions along the two opposite edge sides form the quantum internal loop current, which can generate one big magnetic moment. At these two energy points − 0.17 eV and 0.15 eV the two induced magnetic moments are in opposite signals. [ABSTRACT FROM AUTHOR]

The variation of frequency is a significant indicator of the operation status of rotating machinery. Generally, the frequency is extracted and tracked in real-time based on the vibration signal produced by the rotating machinery. However, various interferences generated from contact sampling and transmitting result in difficulty in obtaining the frequency correctly in real-time from the vibration. To solve this problem, this paper presents an interpolated Kalman filter (IKF) based on the vibration and surface noise signals for real-time frequency tracking. First, the cross-correlation operation is performed on the vibration and surface noise signals sampled synchronously to enhance the energy concentration. After that, a frequency search procedure is carried out to calculate the input of the tracking task. Finally, an IKF-based frequency lock procedure is applied to eliminate the interferences and track the frequency in real time. Besides, a correction procedure is added to prevent the measurement process from tracking the frequency incorrectly. The performance of the proposed method is verified by the experiments based on an ARM-based test bench using standard signals and actual test signals. [ABSTRACT FROM AUTHOR]

Frequency is one of the most significant indicators of power system, which needs accurate estimation to provide a reliable basis for monitoring, control, and protection. Owning to the efficiency, DFT is widely used in the frequency measurement of power systems. However, the inherent defects, that is, the spectral leakage and picket fence effect, decrease the accuracy of DFT. One of the variants, named the windowed phase difference method (WPD), addresses the problem using the phase variation of two windowed sequences in the frequency domain and achieves accurate frequency estimation. Suffering from the ignored negative component and contamination of noise, there are still undesired errors in WPD-based frequency estimation. To reduce the influence of the undesired errors, this paper first analyzes the systematic error of WPD introduced by spectrum leakage and proposes an improved WPD (IWPD) through systematic error compensation to achieve more accurate frequency estimations. Then, the windowing effect on IWPD-based frequency estimation of a noisy signal is studied by deducing the theoretical expression of frequency estimation variance. Finally, the proposed IWPD is validated by extensive computer simulations and practical experiments. [ABSTRACT FROM AUTHOR]

Background: Dynamic N6-methyladenosine (m6A) modification was previously identified as a ubiquitous post-transcriptional regulation that affected mRNA homeostasis. However, the m6A-related epitranscriptomic alterations and functions remain elusive in human cancer. Here we aim to identify the profile and outcome of m6A-methylation in hepatocellular carcinoma (HCC). Results: Using liquid chromatography-tandem mass spectrometry and m6A-immunoprecipitation in combination with high-throughput sequencing, we determined the m6A-mRNA levels in human HCC. Human HCC exhibited a characteristic gain of m6A modification in tandem with an increase of mRNA expression, owing to YTH domain family 2 (YTHDF2) reduction. The latter predicted poor classification and prognosis of HCC patients, and highly correlated with HCC m6A landscape. YTHDF2 silenced in human HCC cells or ablated in mouse hepatocytes provoked inflammation, vascular reconstruction and metastatic progression. Mechanistically, YTHDF2 processed the decay of m6A-containing interleukin 11 (IL11) and serpin family E member 2 (SERPINE2) mRNAs, which were responsible for the inflammation-mediated malignancy and disruption of vascular normalization. Reciprocally, YTHDF2 transcription succumbed to hypoxia-inducible factor-2α (HIF-2α). Administration of a HIF-2α antagonist (PT2385) restored YTHDF2-programed epigenetic machinery and repressed liver cancer. Conclusion: Our results have characterized the m6A-mRNA landscape in human HCC and revealed YTHDF2 as a molecular 'rheostat' in epitranscriptome and cancer progression. [ABSTRACT FROM AUTHOR]

The CRISPR/Cas9 system has been developed as an easy-handle and multiplexable approach for engineering eukaryotic genomes by zygote microinjection of Cas9 and sgRNA, while preparing Cas9 for microinjection is laborious and introducing inconsistency into the experiment. Here, we describe a modified strategy for gene targeting through using oocyte-specific Cas9 transgenic mouse. With this mouse line, we successfully achieve precise gene targeting by injection of sgRNAs only into one-cell-stage embryos. Through comprehensive analysis, we also show allele complexity and off-target mutagenesis induced by this strategy is obviously lower than Cas9 mRNA/sgRNA injection. Thus, injection of sgRNAs into oocyte-specific Cas9 transgenic mouse embryo provides a convenient, efficient and reliable approach for mouse genome editing. [ABSTRACT FROM AUTHOR]

Summary: Monkeys serve as important model species for studying human diseases and developing therapeutic strategies, yet the application of monkeys in biomedical researches has been significantly hindered by the difficulties in producing animals genetically modified at the desired target sites. Here, we first applied the CRISPR/Cas9 system, a versatile tool for editing the genes of different organisms, to target monkey genomes. By coinjection of Cas9 mRNA and sgRNAs into one-cell-stage embryos, we successfully achieve precise gene targeting in cynomolgus monkeys. We also show that this system enables simultaneous disruption of two target genes (Ppar-γ and Rag1) in one step, and no off-target mutagenesis was detected by comprehensive analysis. Thus, coinjection of one-cell-stage embryos with Cas9 mRNA and sgRNAs is an efficient and reliable approach for gene-modified cynomolgus monkey generation. [ABSTRACT FROM AUTHOR]

Highlights: [•] Embryo co-microinjection of Cas9 mRNA and multiple sgRNAs leads to heritable multiple gene modifications. [•] One-step generating multiple immunodeficient mouse models with multiallelic mutations. [•] Optimal concentration of Cas9 mRNA and sgRNAs improved targeting efficiency. [•] Targeting one site with two adjacent sgRNAs increases Cas9:sgRNA-mediated targeting efficiency and bi-allelic mutations. [ABSTRACT FROM AUTHOR]

An amendment to this paper has been published and can be accessed via the original article. [ABSTRACT FROM AUTHOR]

Genetically engineered T cell therapy is emerging as a potent strategy for treating hematological and solid malignancies. Although lentivirus is the most common vector for T cell gene modification, its transduction efficacy remains unsatisfied especially during the manufacturing process. Herein, glycometabolic bioorthogonal chemistry is utilized to establish a highly efficient viral transduction system for human primary T lymphocytes. Azide motifs are anchored on the T cell surface via the intrinsic glycometabolism of exogenous azide–glucose, serving as an artificial ligand for viral binding. The complementary functional moiety dibenzocyclooctyl (DBCO)‐conjugated PEI1.8K (PEI‐DBCO) is then coated on lentiviral surface, which strengthens the virus–T cell interaction through DBCO/azide bioorthogonal chemistry. The results show that the PEI‐DBCO/azide–glucose system effectively facilitates viral binding to T cells and elevates the transduction efficiency of the lentivirus from 20% to 80% without any effect on T cell proliferation and activity. More importantly, the PEI‐DBCO/azide–glucose system significantly doubles the yield of anti‐CD19 chimeric antigen receptor T (CAR‐T) cells and robustly boosts their antitumor capability compared to polybrene‐assisted lentiviral transduction both in vitro and in vivo. Overall, the bioorthogonal PEI‐DBCO/azide–glucose system significantly boosts viral transduction efficacy and exhibits a powerful gene‐manipulating capability in human primary T cells, thereby showing a great potential for clinical‐engineered T lymphocytes manufacture. [ABSTRACT FROM AUTHOR]

The successful use of immune cell checkpoint inhibitors PD-1 and PD-L1, over the past 5 y has raised the concern of using immunotherapy to treat various cancers. Epstein-Barr virus-associated gastric cancer (EBVaGC) exhibits high infiltration of lymphocytes and high amplification of immune-related genes including PD-L1 as distinguished from Epstein-Barr virus-non-associated gastric cancer (EBVnGC). Here, we presume that this PD-1/PD-L1 pathway may hinder the efficacy of adoptive T cell therapy toward EBVaGC. These studies reveal possibility of generating PD-1-disrupted CTL by CRISPR-Cas9 system and demonstrate enhanced immune response of these PD-1-disrupted CTLs to the EBV-LMP2A antigen and superior cytotoxicity to the EBV-positive gastric cancer cell. In addition, when combined with low-dose radiotherapy, these PD-1-disrupted CTLs mediated an impressive antitumor effect in a xenograft mouse model of EBVaGC. Taken together, these studies illustrate PD-1/PD-L1-mediated immune tolerance of EBVaGC and provide a new strategy for targeting immune checkpoints to break the tolerance for the T cell-based adoptive therapy. [ABSTRACT FROM AUTHOR]

Co-injection of zygotes with Cas9 mRNA and sgRNA has been proven to be an efficient gene-editing strategy for genome modification of different species. Genetic engineering in pigs holds a great promise in biomedical research. By co-injection of one-cell stage embryos with Cas9 mRNA and Npc1l1 sgRNA, we achieved precise Npc1l1 targeting in Chinese Bama miniature pigs at the efficiency as high as 100%. Meanwhile, we carefully analyzed the Npc1l1 sgRNA:Cas9-mediated on- and off-target mutations in various somatic tissues and ovaries, and demonstrated that injection of zygotes with Cas9 mRNA and sgRNA is an efficient and reliable approach for generation of gene-modified pigs. [ABSTRACT FROM AUTHOR]