Analysis of swallowing and voice-related quality of life in patients after supracricoid partial laryngectomy.

OBJECTIVE: This study evaluated the swallowing and voice function of laryngeal cancer patients after Supracricoid Partial Laryngectomy(SCPL), and its influence on quality of life to provide a reference for the selection of surgical methods for laryngeal cancer patients. METHODS: Twenty-one patients who received SCPL between April 2015 and November 2021 were included. Each patient's swallowing function and quality of life were assessed through fiberoptic endoscopic examination of swallowing (FEES) and the M.D. Anderson Dysphagia Inventory (MDADI). Fundamental, jitter, shimmer, maximum phonation time (MPT), and voice handicap index-10 (VHI-10) were performed to assess voice function and voice-related quality of life. RESULTS: The results of the FEES of the 21 patients were as follows: the rates of pharyngeal residue after swallowing solid, semiliquid, and liquid food were 0%, 28.57%, and 38.09%, respectively; the rates of laryngeal infiltration after swallowing solid, semiliquid, and liquid food were 0%, 28.57%, and 4.76%, respectively; and aspiration did not occur in any of the patients. In the evaluation of swallowing quality of life, the mean total MDADI score was 92.6 ± 6.32. The voice function evaluation showed that the mean F0, jitter, shimmer, and MPT values were 156.01 ± 120.87 (HZ), 11.57 ± 6.21 (%), 35.37 ± 14.16 (%) and 7.85 ± 6.08 (s), respectively. The mean total VHI-10 score was 7.14 ± 4.84. CONCLUSION: SCPL provides patients with satisfactory swallowing and voice function. The patients in this study were satisfied with their quality of life in terms of swallowing and voice. SCPL can be used as a surgical method to preserve laryngeal function in patients with laryngeal cancer.

Simulated microgravity-induced oxidative stress and loss of osteogenic potential of osteoblasts can be prevented by protection of primary cilia.

Oxidative stress has been considered to be closely related to spaceflight-induced bone loss; however, mechanism is elusive and there are no effective countermeasures. Using cultured rat calvarial osteoblasts exposed to microgravity simulated by a random positioning machine, this study addressed the hypotheses that microgravity-induced shortening of primary cilia leads to oxidative stress and that primary cilium protection prevents oxidative stress and osteogenesis loss. Microgravity was found to induce oxidative stress (as represented by increased levels of reactive oxygen species (ROS) and malondialdehyde production, and decreased activities of antioxidant enzymes), which was perfectly replicated in osteoblasts growing in NG with abrogated primary cilia (created by transfection of an interfering RNA), suggesting the possibility that shortening of primary cilia leads to oxidative stress. Oxidative stress was accompanied by mitochondrial dysfunction (represented by increased mitochondrial ROS and decreased mitochondrial membrane potential) and intracellular Ca2+ overload, and the latter was found to be caused by increased activity of Ca2+ channel transient receptor potential vanilloid 4 (TRPV4), as also evidenced by TRPV4 agonist GSK1016790A-elicited Ca2+ influx. Supplementation of HC-067047, a specific antagonist of TRPV4, attenuated microgravity-induced mitochondrial dysfunction, oxidative stress, and osteogenesis loss. Although TRPV4 was found localized in primary cilia and expressed at low levels in NG, microgravity-induced shortening of primary cilia led to increased TRPV4 levels and Ca2+ influx. When primary cilia were protected by miR-129-3p overexpression or supplementation with a natural flavonoid moslosooflavone, microgravity-induced increased TRPV4 expression, mitochondrial dysfunction, oxidative stress, and osteogenesis loss were all prevented. Our data revealed a new mechanism that primary cilia function as a controller for TRPV4 expression. Microgravity-induced injury on primary cilia leads to increased expression and overactive channel of TRPV4, causing intracellular Ca2+ overload and oxidative stress, and primary cilium protection could be an effective countermeasure against microgravity-induced oxidative stress and loss of osteogenic potential of osteoblasts.

Pseudo-bistability of viscoelastic shells.

Viscoelastic shells subjected to a pressure loading exhibit rich and complex time-dependent responses. Here we focus on the phenomenon of pseudo-bistability, i.e. a viscoelastic shell can stay inverted when pressure is removed, and snap to its natural shape after a delay time. We model and explain the mechanism of pseudo-bistability with a viscoelastic shell model. It combines the small strain, moderate rotation shell theory with the standard linear solid as the viscoelastic constitutive law, and is applicable to shells with arbitrary axisymmetric shapes. As a case study, we investigate the pseudo-bistable behaviour of viscoelastic ellipsoidal shells. Using the proposed model, we successfully predict buckling of a viscoelastic ellipsoidal shell into its inverted configuration when subjected to an instantaneous pressure, creeping when the pressure is held, staying inverted after the pressure is removed, and eventually snapping back after a delay time. The stability transition of the shell from a monostable, temporarily bistable and eventually back to the monostable state is captured by examining the evolution of the instantaneous pressure-volume change relation at different time of the holding and releasing process. A systematic parametric study is conducted to investigate the effect of geometry, viscoelastic properties and loading history on the pseudo-bistable behaviour. This article is part of the theme issue 'Probing and dynamics of shock sensitive shells'.

Large-scale genome editing in plants: approaches, applications, and future perspectives.

As a powerful genome editing technology, CRISPR/Cas is revolutionizing both fundamental research and crop breeding, and has now evolved into large-scale editing tools that are efficient, simple, and programmable. With such CRISPR screening technologies, the numbers of genome-edited crops are rapidly increasing. Here, we describe the general workflow of a CRISPR screen in plants, including the selection of appropriate editors, genome-wide guide RNA design, pooled library construction, massive transformation, and high-throughput genotyping. We also discuss applications for the screening of candidate genes, the optimization of spatiotemporal expression, the evolution of protein activities, and the establishment of genome-wide libraries of knockout mutant. After considering the current challenges and limitations, we finally envision a virus-mediated strategy to improve CRISPR screens.

ESCRT-III component OsSNF7.2 modulates leaf rolling by trafficking and endosomal degradation of auxin biosynthetic enzyme OsYUC8 in rice.

The endosomal sorting complex required for transport (ESCRT) is highly conserved in eukaryotic cells and plays an essential role in the biogenesis of multivesicular bodies and cargo degradation to the plant vacuole or lysosomes. Although ESCRT components affect a variety of plant growth and development processes, their impact on leaf development is rarely reported. Here, we found that OsSNF7.2, an ESCRT-III component, controls leaf rolling in rice (Oryza sativa). The Ossnf7.2 mutant rolled leaf 17 (rl17) has adaxially rolled leaves due to the decreased number and size of the bulliform cells. OsSNF7.2 is expressed ubiquitously in all tissues, and its protein is localized in the endosomal compartments. OsSNF7.2 homologs, including OsSNF7, OsSNF7.3, and OsSNF7.4, can physically interact with OsSNF7.2, but their single mutation did not result in leaf rolling. Other ESCRT complex subunits, namely OsVPS20, OsVPS24, and OsBRO1, also interact with OsSNF7.2. Further assays revealed that OsSNF7.2 interacts with OsYUC8 and aids its vacuolar degradation. Both Osyuc8 and rl17 Osyuc8 showed rolled leaves, indicating that OsYUC8 and OsSNF7.2 function in the same pathway, conferring leaf development. This study reveals a new biological function for the ESCRT-III components, and provides new insights into the molecular mechanisms underlying leaf rolling.

FRET-based fluorescent probe with favorable water solubility for simultaneous detection of SO2 derivatives and viscosity.

The intracellular viscosity is an important parameter of the microenvironment and SO2 is a vital gas signal molecule. At present, some dual-response fluorescence probes for simultaneous measurements of viscosity and SO2 derivatives (HSO3-/SO32-) possessed poor water solubility. In this work, we developed a water-soluble fluorescence probe CIJ (0.0864 g/100 mL of water at 20 °C) for simultaneous measurements of viscosity and SO2 derivatives. CIJ exhibited a sensitive fluorescence enhancement to environmental viscosity from 0.97 to 28.04 cP based on a twisted intramolecular charge transfer mechanism and was applied to effective measurement of viscosity in vitro and in vivo. CIJ could also respond to SO2 derivatives with a low detection limit (44 nM) and a fast response time (5 min) based on the nucleophilic addition reaction. Furthermore, CIJ was applied to monitor SO2 derivatives in ratiometric response manner in living cells.

An imidazo[1,5-α]pyridines-based ratiometric fluorescent probe for sensing sulfur dioxide derivatives in real samples based on a FRET mechanism.

A novel fluorescence resonance energy transfer (FRET)-based ratiometric emission fluorescent probe AT was designed and developed in which the imidazo[1,5-α]pyridine was served as a FRET donor and tricyanofuran (TCF) as the FRET acceptor to detect SO32-/HSO3- based on the Michael addition reaction. Probe AT had a high energy transfer efficiency (95%) and a large pseudo-Stokes shift (259 nm) in EtOH/PBS buffer (5/5, v/v). It also possessed good selectivity and quick response to SO32-/HSO3-. There was good linearity between the ratio of fluorescence intensity (F499/F645) and the concentrations of SO32-/HSO3- in the ranges of 1.5-7.5 µM and 9-20 µM, with calculated detection limits (LOD) of 55 nM. In addition, the probe could also detect the concentrations of SO32-/HSO3- in real samples such as environmental water and sugar, allowing the probe to be used in a variety of applications.

Dwarf and High Tillering1 represses rice tillering through mediating the splicing of D14 pre-mRNA.

Strigolactones (SLs) constitute a class of plant hormones that regulate many aspects of plant development, including repressing tillering in rice (Oryza sativa). However, how SL pathways are regulated is still poorly understood. Here, we describe a rice mutant dwarf and high tillering1 (dht1), which exhibits pleiotropic phenotypes (such as dwarfism and increased tiller numbers) similar to those of mutants defective in SL signaling. We show that DHT1 encodes a monocotyledon-specific hnRNP-like protein that acts as a previously unrecognized intron splicing factor for many precursor mRNAs (pre-mRNAs), including for the SL receptor gene D14. We find that the dht1 (DHT1I232F) mutant protein is impaired in its stability and RNA binding activity, causing defective splicing of D14 pre-mRNA and reduced D14 expression, and consequently leading to the SL signaling-defective phenotypes. Overall, our findings deepen our understanding of the functional diversification of hnRNP-like proteins and establish a connection between posttranscriptional splicing and SL signaling in the regulation of plant development.

Two simple but effective turn-on benzothiazole-based fluorescent probes for detecting hydrogen sulfide in real water samples and HeLa cells.

Two simple turn-on fluorescent probes, containing a benzothiazole and the 2,4-dinitrobenzenesulfonyl group, were designed for detecting H2S. Two probes exhibited good selectivity and high sensitivity, which were applied to detect the H2S in real water samples. Probe P2 with a positive charge had better solubility than probe P1 in water; therefore, probe P2 was successfully applied to detect both the endogenous and exogenous H2S in lysosomes of living HeLa cells.

Metal-Free Catalyzed Synthesis of Fluorescent Indolizine Derivatives.

A mild and high efficient method to prepare indolizines by two-component reaction with the acid as the catalyst was developed. In this reaction, a new ring efficiently formed in one-step reaction. A wide range of substrates could be applied and the desired products were obtained in 8-95% yields under metal-free conditions. Different indolizine derivatives (compounds 3a-3n) were synthesized by general conditions and microwave irradiation conditions, and compound 3a gave the best results with an isolated yield of 95% and 82%, respectively. The structures of synthesized compounds were characterized by spectral analysis, and compound 3m was confirmed by single crystal X-ray analysis. UV-vis absorption and fluorescence properties of these compounds were correlated with substituent groups on indolizine rings.

Establishment of a Stable ß-Casein Protein-Secreted Laoshan Dairy Goat Mammary Epithelial Cell Line.

Mammary epithelial cells are widely used as models in mastitis research and as tools for mammalian bioreactors; however, the short lifespan of these cells limits their utility. Several mammal epithelial cell line models have been established; however, the secretion capacity and the bacterial sensitivity of these lines have not been effectively evaluated. In this study, a stable immortalized goat mammary epithelial cell (GMEC) line was constructed by transfection with the SV40 gene. The monoclonal cells were then passaged through more than 50 generations after puromycin selection. The GMEC line was evaluated by reverse transcriptase polymerase chain reaction, the cell cycle, karyotype analysis, detection of apoptosis, Western blotting, and ß-casein (CSN2) inducible assays. The GMEC line had a strong proliferation capacity relative to the primary GMECs. GMECs had the same karyotype as the primary cells. The GMEC lines maintained basic biological properties and had estrogen, prolactin, and progesterone receptors as same the primary cells. Additionally, the cells and the cell line could synthesize and secrete ß-casein proteins. Finally, the rate of apoptosis of the transfected cells suggested that the cell line could provide a useful tool for signal research and mammary gland bioreactors.

Ubiquitin Specific Protease 15 Has an Important Role in Regulating Grain Width and Size in Rice.

Ubiquitination and deubiquitination are reversible processes that play crucial roles in regulating organ size in plants. However, information linking deubiquitination and seed size in rice (Oryza sativa) is limited. Here, we characterized a dominant large-grain mutant, large grain1-D (lg1-D), with a 30.8% increase in seed width and a 34.5% increase in 1,000-grain weight relative to the wild type. The lg1-D mutant had more cells oriented in the lateral direction of the spikelet hull compared with the wild type. Map-based cloning showed that LG1 encodes a constitutively expressed ubiquitin-specific protease15 (OsUBP15) that possesses deubiquitination activity in vitro. Loss-of-function and down-regulated expression of OsUBP15 produced narrower and smaller grains than the control. A set of in vivo experiments indicated that the mutant Osubp15 had enhanced protein stability relative to wild-type OsUBP15. Further experiments verified that OsDA1 directly interacted with OsUBP15. Genetic data indicated that OsUBP15 and GRAIN WIDTH 2 (GW2) were not independent in regulating grain width and size. In summary, we identified OsUBP15 as a positive regulator of grain width and size in rice and provide a promising strategy for improvement of grain yield by pyramiding OsUBP15 and gw2.

DELAYED HEADING DATE1 interacts with OsHAP5C/D, delays flowering time and enhances yield in rice.

Heading date is an important agronomic trait affecting crop yield. The GRAS protein family is a plant-specific super family extensively involved in plant growth and signal transduction. However, GRAS proteins are rarely reported have a role in regulating rice heading date. Here, we report a GRAS protein DHD1 (Delayed Heading Date1) delays heading and enhances yield in rice. Biochemical assays showed DHD1 physically interacts with OsHAP5C/D both in vitro and in vivo. DHD1 and OsHAP5C/D located in the nucleus and showed that rhythmic expression. Both DHD1 and OsHAP5C/D affect heading date by regulating expression of Ehd1. We propose that DHD1 interacts with OsHAP5C/D to delay heading date by inhibiting expression of Ehd1.

Transesterification of para-hydroxybenzoic acid esters (parabens) in the activated sludge.

Hydrolysis is generally considered as the main pathway for the degradation of ester-type pollutants in aquatic environments. In this study, we found that when methanol or ethanol presented as the external carbon in the activated sludge, transesterification is very important for the degradation of para-hydroxybenzoic acid esters (parabens). In the activated sludge solutions with 1% methanol added, contribution of transesterification to the degradation of the propyl substituted paraben (PrP) and ethyl substituted paraben (EtP) accounted for 46% and 83%, respectively, in the early stage of the reaction. This indicates that in aquatic environments with alcohols presence, parabens prefer to form small molecule homologues than hydrolysis to acid. The predominant transesterification in the activated sludge is related to enzyme preference. Amano lipase from Pseudomonas fluorescens was verified to catalyze hydrolysis and transesterification of parabens, while the latter was dominant in water solution with 1% methanol or ethanol. Considering the common application of small molecular alcohols as the external carbon sources in wastewater treatment plants, transesterification might be an important pathway for the degradation of parabens pollutants in these engineering aquatic environments.

Three-Dimensional Reduced Graphene Hydrogels Using Various Carbohydrates for High Performance Supercapacitors.

In present work, three-dimensional (3D) reduced graphene hydrogels (RGHs) are prepared through an efficient and facile strategy by employing three types of carbohydrates (glucose, fructose and sucrose) as reducing agents in aqueous solution of graphene oxide (GO) with ammonia. The formation of RGHs could be confirmed by X-ray powder diffraction (XRD), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FT-IR). The structures and porosity were characterized by field emission scanning electron microscopy (FESEM), high resolution transmission electron microscopy (HRTEM) and N2 sorption experiments. Benefiting from the abundant porous architectures as fast ionic channels for electrochemical energy storage, the prepared RGHs exhibited a high specific capacitance up to 153.5, 145.0 and 150.3 F g−1 at 0.3 A g−1 for FRGHs (fructose), GRGHs (glucose) and SRGHs (sucrose), which can be maintained for 61.4, 61.5 and 46.9% as the discharging current density was increased up to 20 A g−1. Moreover, it also showed that the electrode based on RGHs has good electrochemical stability and high degree of reversibility in the charge/discharge cycling test.

Occurrence and Profile Characteristics of the Pesticide Imidacloprid, Preservative Parabens, and Their Metabolites in Human Urine from Rural and Urban China.

Knowledge of human exposure to imidacloprid, the most extensively used insecticide, and para-hydroxybenzoic acid esters (parabens), the most extensively used preservative, is insufficient. In this study, 295 urine samples collected from subjects in rural and urban areas in China were analyzed for imidacloprid and four parabens (namely, methyl paraben, ethyl paraben, propyl paraben, and butyl paraben) as well as their major metabolites (namely, 6-chloronicotinic acid (6-ClNA) and para-hydroxybenzoic acid (p-HB)). Imidacloprid was detected in 100% of the urine samples from rural Chinese subjects and 95% of the urine samples from urban Chinese subjects. Concentrations of urinary imidacloprid detected in rural Chinese subjects (geometric mean (GM) = 0.18 ng/mL) were slightly higher than those detected in urban Chinese subjects (GM = 0.15 ng/mL) when the effect of pesticide spraying was excluded. However, concentrations of urinary imidacloprid detected in rural adults increased significantly in the subsequent days of pesticide spraying (GM = 0.62 ng/mL), which could return to the normal levels within 3 days. In contrast, concentrations of urinary parabens detected in rural Chinese subjects (GM = 6.90 ng/mL) were lower than that in urban Chinese subjects (GM = 30.5 ng/mL). In addition, the metabolism characteristics of imidacloprid to 6-ClNA and parabens to p-HB were discussed preliminarily.