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Background: The intestinal mucosal immune system, renowned for its precise and sensitive regulation, can provide comprehensive and effective protection for the body, among which the ileum is a critical induction site for regulating mucosal immune homeostasis. Moniezia benedeni parasitizes the small intestine of sheep and can cause serious pathological damage or even death to the host when the infection is severe. In this study, 5 sheep infected with Moniezia benedeni were selected as the infected group, and 5 uninfected sheep were selected as the control group. The ileal transcriptome profile characteristics of Moniezia benedeni infection were analyzed based on RNA-seq sequencing technology, aiming to lay a foundation for further exploring the perception mechanism of sheep intestines to Moniezia benedeni infection and formulating effective prevention and control strategies., Results: The results showed that a total of 3,891 differentially expressed genes (DEGs) were detected in the ileum tissues of sheep between the infected and control groups with 2,429 up-regulated genes and 1,462 down-regulated genes. GO and KEGG pathway enrichment analysis of differential genes, as well as Clue GO analysis showed that differential genes were significantly enriched in immune and metabolic-related biological processes and signaling pathways. Particularly, in immune-related signaling pathways, the B cell receptor signaling pathway was significantly down-regulated, while in metabolic regulation related signaling pathways, Bile secretion, Fat digestion and absorption and Vitamin digestion and absorption were notably up-regulated. On this basis, the differential core genes related to immune metabolism were verified by qRT-PCR method. The results showed that OVAR, CD3E, CD8A, CD4 and CD28 were significantly up-regulated (P < 0.05), while CIITA, BLNK, BCL6 and CD79A were significantly down-regulated (P < 0.05), which were consistent with transcriptome sequencing data., Conclusions: The results demonstrated that Moniezia benedeni infection significantly affected the immune and metabolic processes in sheep ileum, particularly, it significantly inhibited the activation process of host B cells, and also led to an overactive function of bile acid metabolism. This finding provides a solid foundation for further elucidating the response mechanism of Peyer's patches in sheep ileum to Moniezia tapeworm infection., (© 2024. The Author(s).)

The insufficient abundance of electron acceptors for ammonia during electron transfer in constructed wetlands (CWs) results in low nitrification rates. This study developed a green, low-carbon CWs enhanced by a bio-electrochemical systems (BESs-CWs) to achieve efficient ammonia (NH 4 + -N) removal. Electrode enhancement significantly promoted NH 4 + -N removal. Compared with traditional CWs, the average removal efficiency of NH 4 + -N in the BESs-CWs increased from 62.9 % to 90.6 %. The intermittent voltage driven by the photovoltaic power system caused minimal plant stress. However, electrode enhancement significantly affected microbial communities involved in short-path nitrification and denitrification within the biofilm. Specifically, the removal rate of NH 4 + -N by BESs-CWs under electrode enhancement was increased by 27.7 % compared to traditional CWs, enhancing the electron output of NH 4 + -N in the BESs-CWs. This system provides a method of ammonia nitration for CWs under poor electron acceptor conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Emerging evidence suggests that circular RNAs (circRNAs) are involved in the regulation of tumourigenesis and progression of a variety of malignant tumours. In this study, we aimed to identify laryngeal squamous cell carcinoma (LSCC)-specific circRNAs and explore their biological functions and underlying molecular mechanisms. Employing microarray and qRT-PCR, hsa&#95;circ&#95;0000825 was found to be significantly increased in LSCC tissues versus para-cancerous tissues. High hsa&#95;circ&#95;0000825 expression was positively associated with advanced clinical stages, lymph node metastasis, and poor survival. Furthermore, the overexpression of hsa&#95;circ&#95;0000825 in TU177 and AMC-HN-8 cells promoted cell proliferation. Transwell assays showed enhanced migration and invasion of TU177 and AMC-HN-8 cells upon overexpression of hsa&#95;circ&#95;0000825. Conversely, the knockdown of hsa&#95;circ&#95;0000825 had the opposite effect. Xenograft tumours in BALB/c nude mice derived from hsa&#95;circ&#95;0000825-overexpressed TU177 cells showed greater volume and weight than those derived from control TU177 cells. Mechanistically, nuclear-cytoplasmic fractionation assay confirmed that hsa&#95;circ&#95;0000825 was mainly located in the cytoplasm of TU177 and AMC-HN-8 cells. The AGO2-RNA immunoprecipitation (RIP) assay revealed that hsa&#95;circ&#95;0000825 was significantly enriched in the AGO2-precipitated complex in both TU177 and AMC-HN-8 cells, suggesting that this circRNA may function via a competitive endogenous RNA (ceRNA) mechanism. Next, bioinformatics analysis, biotinylated-oligo pull-down assay and dual-luciferase reporter assay verified that miR-766 could be sponged by hsa&#95;circ&#95;0000825 and also target 3'UTR of HOXD10 mRNA. Moreover, miR-766 was shown to be involved in the pro-oncogenic effect of hsa&#95;circ&#95;0000825. This occurred via the mediation of hsa&#95;circ&#95;0000825-enhanced HOXD10 mRNA by the ceRNA mechanism in TU177 and AMC-HN-8 cells. Besides, RNA-binding protein (RBP) ELAVL1 interacted with hsa&#95;circ&#95;0000825 in TU177 and AMC-HN-8 cells, as revealed through bioinformatics analysis, biotinylated-oligo pull-down assays, and RIP assays. ELAVL1 knockdown decreased cell proliferation by 38 % and 34 % in hsa&#95;circ&#95;0000825-overexpressed TU177 and AMC-HN-8 cells ( P  < 0.05). Similarly, ELAVL1 was involved in the pro-migration and pro-invasion effects of hsa&#95;circ&#95;0000825 overexpression. In addition, comprehensive analysis of mRNA-seq in hsa&#95;circ&#95;0000825-overexpressed TU177 cells, as well as catRAPID and TCGA databases, suggested that ITGB2, HOXD10, and MTCL1 might be crucial downstream target mRNAs of ELAVL1 in LSCC, participating in the hsa&#95;circ&#95;0000825-ELAVL1 axis pro-oncogenic effect. Taken together, hsa&#95;circ&#95;0000825 plays a pro-oncogenic role in LSCC via the miR-766/HOXD10 axis and ELAVL1 and may serve as a promising specific biomarker and therapeutic target for LSCC., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Limited organic carbon in drinking water constrains the removal of nitrate‑nitrogen (NO 3 - -N) via aerobic denitrification. This paper reports the use of silica-coated nano zero-valent iron (nZVI@SiO 2 ) as a stable and sustainable electron donor to enhance aerobic denitrification. The nZVI@SiO 2 , synthesized via a one-step method, was resistant to oxidation and exhibited excellent stability. In conjunction with aerobic denitrifying bacteria, nZVI@SiO 2 achieved NO 3 - -N and total nitrogen TN removal efficiencies of 90.64 % and 80.94 %, respectively. This represents an increase of 24.15 % in the efficiency of TN removal compared with that of the nZVI system. The activity of the nZVI system diminished gradually after just three cycles, whereas nZVI@SiO 2 maintained NO 3 - -N and TN removal efficiencies of 89.33 % and 78.08 %, respectively, after four cycles, respectively, indicating its sustainable ability to enhance aerobic denitrification. Cyclic voltammetry and electrochemical impedance spectroscopy demonstrated enhanced electron transfer efficiency of nZVI@SiO 2 . Furthermore, nZVI@SiO 2 significantly promoted the activity of the electron transfer system, ATP levels, nitrate/nitrite reductase activity, contents of complexes I and III, and extracellular polymeric substances. nZVI@SiO 2 significantly enhanced electron generation, transfer, and consumption during biological denitrification by functioning as both an electron donor and mediator. The findings implicate nZVI@SiO 2 as a means to enhance nitrogen removal by aerobic denitrifying microorganisms in oligotrophic water via sustained donation of electrons., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Reducing nitrate contamination in drinking water has become a critical issue in urban water resource management. Here a novel oligotrophic aerobic denitrifying bacterium, Pelomonas puraquae WJ1, was isolated and purified from artificial lake sediments. For the first time, excellent aerobic denitrification capabilities were demonstrated. At a carbon-to‑nitrogen ratio of 5.0, strain WJ1 achieved 100.0 % nitrate removal and 84.92 % total nitrogen removal within 24 h, with no nitrite accumulation. PCR amplification and sequencing confirmed the presence of the denitrification genes napA, nirS, and nosZ in the strain. The nitrogen balance demonstrated that approximately 74.95 % of the initial nitrogen was eliminated as gaseous products under aerobic conditions. Furthermore, carbon balance analysis showed that most electron donors from strain WJ1 were directed towards oxygen, with limited availability for nitrate reduction. A combination of bio-ECO analysis and network modeling indicated that strain WJ1 has robust metabolic capabilities for diverse carbon sources and exhibits high adaptability to complex carbon environments. Overall, Pelomonas puraquae WJ1 removed approximately 45.89 % of the nitrates in raw water, demonstrating significant potential for practical applications in oligotrophic denitrification., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Limonium bicolor, known horticulturally as sea lavender, is a typical recretohalophyte with salt glands in its leaf epidermis that secrete excess Na+ out of the plant. Although many genes have been proposed to contribute to salt gland initiation and development, a detailed analysis of alternative splicing, alternative polyadenylation patterns, and long non-coding RNAs (lncRNAs) has been lacking. Here, we applied single-molecule long-read mRNA isoform sequencing (Iso-seq) to explore the complexity of the L. bicolor transcriptome in leaves during salt gland initiation (stage A) and salt gland differentiation (stage B) based on the reference genome. We identified alternative splicing events and the use of alternative poly(A) sites unique to stage A or stage B, leading to the hypothesis that they might contribute to the differentiation of salt glands. Based on the Iso-seq data and RNA in situ hybridization of candidate genes, we selected the lncRNA Btranscript&#95;153392 for gene editing and virus-induced gene silencing to dissect its function. In the absence of this transcript, we observed fewer salt glands on the leaf epidermis, leading to diminished salt secretion and salt tolerance. Our data provide transcriptome resources for unraveling the mechanisms behind salt gland development and furthering crop transformation efforts towards enhanced survivability in saline soils., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Heptafluoro-iso-butyronitrile ( i -C 3 F 7 CN) represents a feasible eco-friendly replacement gas for the most potent greenhouse gas sulfur hexafluoride in various high-voltage power transmission equipment. The reaction mechanisms for the in situ synthesis of i -C 3 F 7 CN from heptafluoro-iso-butyramide [ i -C 3 F 7 C(O)NH 2 ] in the presence of trifluoroacetic anhydride (TFAA) and pyridine (Py) in dimethylformamide solution have been studied within density functional theory with M06-2X exchange-correlation functional with the 6-311++G(d,p) basis set and the high-level ab initio complete basis set quadratic CBS-QB3 method. It is revealed that the unimolecular dehydration of i -C 3 F 7 C(O)NH 2 can be catalyzed efficiently by TFAA in terms of both kinetic and thermodynamic aspects, producing i -C 3 F 7 CN and trifluoroacetic acid (TFA). Furthermore, Py is capable of reducing the energy barrier of the rate-determining step through hydrogen abstraction to form pyridinium hydrogen. The synergic effect of the TFAA/Py co-catalyst plays a pivotal role in the production of i -C 3 F 7 CN as the Gibbs free energy barrier can be lowered by more than 40 kcal/mol with the ratio of TFAA:2Py, in accordance with the experimental observation. The present theoretical work provides new insights into the rational design on the novel catalysts for large-scale synthesis of the perfluorinated nitriles.

Background: Glycolysis is an indispensable process for tumor cell,but the effect of glycolysis on the prognosis and immune cell infiltration of head and neck squamous cell carcinoma is not clear., Methods: Based on RNA bulk and single cell RNA sequencing data of head and neck squamous cell carcinoma from The Cancer Genome Atlas(TCGA) and GSE195832, the effect of glycolysis level on immune cell infiltration was analyzed. Then, we obtained the prognostic genes related to glycolysis through survival analysis to construct prognostic risk signature. Our sample and GSE65858 datasets are used as external verification datasets to verify the validity of the signature. Finally, we used Western blot and cell function assays to determine the relationship between risk genes and glycolysis and the function of prognostic genes., Result: The level of glycolysis was related to the prognosis of head and neck tumors (P = 0.0044). The results of immune infiltration analysis of TCGA database showed that high level glycolysis subgroup had less infiltration of macrophages, T cells and monocytes. Results of single cell sequencing analysis validates the above results. Additionally, Five risk genes(MUCL1,TRIML2,RAB3B,SPINK6,IGSF11) were selected to construct signature.Risk score was an independent prognostic factor(P < 0.01). The external validation set also shows the same result. In vitro functional and Western blot assays confirmed that the above five genes affect tumor function and related to the process of glycolysis., Conclusion: Glycolysis-related risk signatures can be used to predict the prognosis and immune infiltration of head and neck squamous cell carcinoma., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024. Published by Elsevier Inc.)

Introduction: Immunoglobulin G (IgG) is important in mediating humoral immunity and in the maintenance of immune homeostasis in the intestinal mucosa. Oregano essential oil (OEO) is a natural herbal extract that possesses antimicrobial, antioxidant, anti-inflammatory, and immunomodulatory properties. As the effects of OEO on intestinal mucosal immunity in Holstein dairy bulls remained unclear, we investigated the effect of dietary supplementation of OEO on IgG levels and IgG + cells residing in the intestinal tract in Holstein dairy bulls., Methods: Twelve Holstein bulls in good health of approximately 10 months of age were selected for the experiment and randomly equally divided into two groups. The control (CK) group was fed a basal ration, and in the OEO group, the basal ration was supplemented with OEO (20 g/head/day). After 300 days of feeding, tissue samples of the jejunum, ileum, and colon of the bulls in each group were collected for histopathological analysis, immunohistochemistry, and enzyme-linked immunosorbent assays, respectively., Results: The jejunum, ileum, and colon of bulls in the CK group had obvious pathological damage, whereas the structure of each intestinal segment was clear and intact. In the OEO group, pathological damage was significantly reduced. IgG + plasma cells were diffusely distributed in the lamina propria of the jejunum, ileum, and colon in the CK and OEO groups, with no significant difference between the groups. OEO supplementation significantly reduced the number of IgG+ plasma cells in each intestinal segment, with the highest decrease rate being noted for the ileum (22.87%), followed by the colon (19.45%) and jejunum (8.52%). ELISA test results and immunohistochemical results were mutually verified. The change in IgG content was consistent with the trend of change in the number of IgG + plasma cells., Discussion: Our findings suggest that OEO supplementation does not alter the diffuse spatial distribution of IgG + plasma cells in the intestines of Holstein dairy bulls, but lowers immunoglobulin levels to normal levels, significantly reduces intestinal damage, and may enhance mucosal immune defence barrier function by inhibiting inflammatory reactions., Competing Interests: QC was employed by Jing Chuan Xu Kang Food Co., Ltd. ZZ was employed by Gansu Huarui Agriculture Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Xu, Zhang, Kong, Wang, Pan, Shi, Liu, He, Ma, Cheng, Zhang and Lei.)

Background: The neuroimmune network plays a crucial role in regulating mucosal immune homeostasis within the digestive tract. Synaptosome-associated protein 25 (SNAP-25) is a presynaptic membrane-binding protein that activates ILC2s, initiating the host's anti-parasitic immune response., Methods: To investigate the effect of Moniezia benedeni (M. benedeni) infection on the distribution of SNAP-25 in the sheep's small intestine, the recombinant plasmid pET-28a-SNAP-25 was constructed and expressed in BL21, yielding the recombinant protein. Then, the rabbit anti-sheep SNAP-25 polyclonal antibody was prepared and immunofluorescence staining was performed with it. The expression levels of SNAP-25 in the intestines of normal and M. benedeni-infected sheep were detected by ELISA., Results: The results showed that the SNAP-25 recombinant protein was 29.3 KDa, the titer of the prepared immune serum reached 1:128,000. It was demonstrated that the rabbit anti-sheep SNAP-25 polyclonal antibody could bind to the natural protein of sheep SNAP-25 specifically. The expression levels of SNAP-25 in the sheep's small intestine revealed its primary presence in the muscular layer and lamina propria, particularly around nerve fibers surrounding the intestinal glands. Average expression levels in the duodenum, jejunum, and ileum were 130.32 pg/mg, 185.71 pg/mg, and 172.68 pg/mg, respectively. Under conditions of M. benedeni infection, the spatial distribution of SNAP-25-expressing nerve fibers remained consistent, but its expression level in each intestine segment was increased significantly (P < 0.05), up to 262.02 pg/mg, 276.84 pg/mg, and 326.65 pg/mg in the duodenum, jejunum, and ileum, and it was increased by 101.06%, 49.07%, and 89.16% respectively., Conclusions: These findings suggest that M. benedeni could induce the SNAP-25 expression levels in sheep's intestinal nerves significantly. The results lay a foundation for further exploration of the molecular mechanism by which the gastrointestinal nerve-mucosal immune network perceives parasites in sheep., (© 2024. The Author(s).)

Background: There are no direct comparisons across different endoscopic therapies for gastroesophageal reflux disease (GERD). This study aimed to evaluate the relative effects of different endoscopic therapies in GERD., Methods: Five databases were searched until August 2023 for randomized controlled trials (RCTs) that compared the efficacy of endoscopic band ligation (EBL), Stretta, endoscopic fundoplication (transoral incisionless fundoplication [TIF], endoscopic full-thickness plication [EFTP], and EndoCinch plication procedure [EndoCinch, CR BARD, Billerica, Mass., USA]), or proton pump inhibitors (PPIs)/sham procedure for GERD. Bayesian network meta-analysis was performed., Results: A total of 19 trials comprising 1181 patients were included. EBL (mean difference [MD], -7.75; 95% credible interval [CrI], -13.90 to -1.44), Stretta (MD, -9.86; 95% CrI, -19.05 to -0.58), and TIF (MD, -12.58; 95% CrI, -20.23 to -4.91) all significantly improved patients' health-related quality of life score with equivalent efficacy compared with PPIs. TIF and EBL achieved equivalent efficacy in reducing PPIs utility (risk ratio [RR], 0.66; 95% CrI, 0.40-1.05) and both were significantly superior to other endoscopic interventions (Stretta, EFTP, and EndoCinch). Besides, EBL and TIF also could significantly decrease the esophagitis incidence compared with PPIs (EBL [RR, 0.34; 95% CrI, 0.22-0.48] and TIF [RR, 0.38; 95% CrI, 0.15-0.88]). In terms of lower esophageal sphincter (LES) pressure, only TIF could significantly increase the LES pressure (MD, 6.53; 95% CrI, 3.65-9.40) to PPIs. In contrast, TIF was inferior to PPIs in decreasing esophageal acid exposure (MD, 2.57; 95% CrI, 0.77-4.36)., Conclusion: Combining the evidence, EBL and TIF may have comparable efficacy and both might be superior to Stretta, EFTP, or EndoCinch in GERD treatment., (Copyright © 2024 Society for Surgery of the Alimentary Tract. Published by Elsevier Inc. All rights reserved.)

Salt gland is an epidermal Na+ secretory structure that enhances salt resistance in the recretohalophyte sea lavender (Limonium bicolor). To elucidate the salt gland development trajectory and related molecular mechanisms, we performed single-cell RNA sequencing of L. bicolor protoplasts from young leaves at salt gland initiation and differentiation stages. Dimensionality reduction analyses defined 19 transcriptionally distinct cell clusters, which were assigned into 4 broad populations-promeristem, epidermis, mesophyll, and vascular tissue-verified by in situ hybridization. Cytokinin was further proposed to participate in salt gland development by the expression patterns of related genes and cytological evidence. By comparison analyses of Single-cell RNA sequencing with exogenous application of 6-benzylaminopurine, we delineated 5 salt gland development-associated subclusters and defined salt gland-specific differentiation trajectories from Subclusters 8, 4, and 6 to Subcluster 3 and 1. Additionally, we validated the participation of TRIPTYCHON and the interacting protein Lb7G34824 in salt gland development, which regulated the expression of cytokinin metabolism and signaling-related genes such as GLABROUS INFLORESCENCE STEMS 2 to maintain cytokinin homeostasis during salt gland development. Our results generated a gene expression map of young leaves at single-cell resolution for the comprehensive investigation of salt gland determinants and cytokinin participation that helps elucidate cell fate determination during epidermis formation and evolution in recretohalophytes., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Mineral oils and synthetic and natural esters are the predominant insulating liquids in electrical equipment. Structure-activity relationship models to predict the key properties of pure insulating liquids, including pulse breakdown strengths, AC breakdown voltages, dielectric constants, flash points, and kinematic viscosities, have been proposed for the first time. Dependence of the specific properties on the molecular structures has been illustrated quantitatively in terms of surface area, statistical total variance, and average deviation of positive and negative electrostatic potentials, as augmented by molecular weight, volume, and ovality. Moreover, the individual contribution of the functional groups to viscosity has been revealed by an additive approach. The predicted properties are in good agreement with the experimental data. The present theoretical work provides new insights on the development of novel dielectric fluids.

The rapid recombination of photogenerated electrons and holes greatly limits the efficiency of photocatalyst based on semiconductor. In order to address this issue, we predicted a novel ferroelectric polarized heterojunction photocatalyst, CdS/In 2 Se 3 , which enables the spontaneous overall water splitting reaction. The CdS/In 2 Se 3 heterojunction exhibits a band-edge staggered alignment and it is easy to form a direct Z-scheme charge transfer pathway. Besides, the built-in electric field ( E int ) in the CdS/In 2 Se 3 heterojunction promoted the charge transfer of CdS/In 2 Se 3 , leading to an improved separating efficiency of photo-generated carriers. Moreover, the vertical intrinsic polarized electric field ( E p ) not only alters the position of the band edge but also reduces the bandgap limitations commonly associated with photocatalytic materials. Furthermore, the CdS/In 2 Se 3 heterojunctions demonstrate separate catalytic activity for the hydrogen evolution reaction (HER) on the surface of the CdS monolayer and oxygen evolution reaction (OER) on the surface of In 2 Se 3 , respectively. Notably, the CdS/In 2 Se 3 -down configuration enables spontaneous photocatalytic water splitting in pH = 7, while the CdS/In 2 Se 3 -up configuration efficiently facilitates the HER process. This study highlights the significant advantages of CdS/In 2 Se 3 heterojunctions as photocatalytic materials, offering unique insights into the development and research of this promising heterojunction architecture.

Purpose: To analyze and compare clinical research trends and hot topics in allergic rhinitis (AR) and asthma and provide valuable theoretical data and references for future research., Methods: Clinical studies focusing on AR or asthma published from 2013 to 2023 were retrieved from the Web of Science Core Collection. Eligible articles were screened and analyzed using bibliometrics from multiple indicators., Results: A total of 261 eligible articles on AR and 991 qualified articles on asthma were screened. The following bibliometric analyses identified the Journal of Allergy and Clinical Immunology as the most influential publication on AR and asthma and proved the significant contributions of Harvard University in clinical studies on AR and asthma. The analyses also revealed that the top ten prolific authors for AR were from China, the United Kingdom, Japan, and Germany, whereas the top ten productive authors for asthma were mainly from the USA. Collaborations among countries for AR were relatively concentrated in the Occident, whereas international cooperation on asthma was mainly achieved by the Occident and certain Eastern countries., Conclusions: This study compared and analyzed the current status and evolution of AR and asthma-related clinical research using bibliometric analysis. Additionally, the study comprehensively summarized the impactful authors, institutions, and countries, and revealed the replacement and evolution of hotspots., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

The recretohalophyte Limonium bicolor thrives in high-salinity environments because salt glands on the above-ground parts of the plant help to expel excess salt. Here, we characterize a nucleus-localized C3HC4 (RING-HC)-type zinc finger protein of L. bicolor named  RING  ZINC  FINGER PROTEIN  1 (LbRZF1). LbRZF1 was expressed in salt glands and in response to NaCl treatment. LbRZF1 showed no E3 ubiquitin ligase activity. The phenotypes of overexpression and knockout lines for LbRZF1 indicated that LbRZF1 positively regulated salt gland development and salt tolerance in L. bicolor. lbrzf1 mutants had fewer salt glands and secreted less salt than did the wild-type, whereas LbRZF1-overexpressing lines had opposite phenotypes, in keeping with the overall salt tolerance of these plants. A yeast two-hybrid screen revealed that LbRZF1 interacted with LbCATALASE2 (LbCAT2) and the transcription factor LbMYB113, leading to their stabilization. Silencing of LbCAT2 or LbMYB113 decreased salt gland density and salt tolerance. The heterologous expression of LbRZF1 in Arabidopsis thaliana conferred salt tolerance to this non-halophyte. We also identified the transcription factor LbMYB48 as an upstream regulator of LbRZF1 transcription. The study of LbRZF1 in the regulation network of salt gland development also provides a good foundation for transforming crops and improving their salt resistance., (© 2024 The Authors. Journal of Integrative Plant Biology published by John Wiley & Sons Australia, Ltd on behalf of Institute of Botany, Chinese Academy of Sciences.)

NAC transcription factors regulate plant growth, development, and stress responses. However, the number, types, and biological functions of Limonium bicolor LbNAC genes have remained elusive. L. bicolor secretes excessive salt ions through salt glands on its stems and leaves to reduce salt-induced damage. Here, we identified 63 NAC members (LbNAC1-63) in L. bicolor, which were unevenly distributed across eight chromosomes. Cis-elements in the LbNAC promoters were related to growth and development, stress responses, and phytohormone responses. We observed strong colinearity between LbNACs and GmNACs from soybean (Glycine max). Thus, LbNAC genes may share similar functions with GmNAC genes. Expression analysis indicated that 16 LbNAC genes are highly expressed in roots, stems, leaves, and flowers, whereas 17 LbNAC genes were highly expressed throughout salt gland development, suggesting that they may regulate this developmental stage. Silencing LbNAC54 in L. bicolor decreased salt gland density, salt secretion from leaves, and overall salt tolerance. In agreement, genes related to salt gland development were significantly downregulated in LbNAC54-silenced lines. Our findings shed light on LbNAC genes and help elucidate salt gland development and salt secretion in L. bicolor. Our data also provide insight into NAC functions in halophytes., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Tetrafluoropropyne (C 3 F 4 ) is a potential dielectric in various electrical insulating equipment to replace the most potent industrial greenhouse gas, sulfur hexafluoride. Atmospheric oxidation of C 3 F 4 by OH radicals in the presence of molecular O 2 has been investigated theoretically in order to clarify the lifetime and degradation products at mechanistic and kinetic aspects. Energetic minimum-energy pathways for the C 3 F 4 + OH/O 2 reactions were calculated in detail using various theoretical methods including density functional M06-2X and CCSD for geometries, CBS-QB3, CCSD(T), and multireference RS2 with extrapolation to the complete basis-set limit for energies. It has been demonstrated that the C 3 F 4 + OH reaction takes place via the bifurcated C-O addition/elimination routes leading to CF 3 C(OH)═CF and CF 3 C═C(OH)F radical adducts, where the latter is more preferable in view of the difference in barrier heights (1.3 vs 0.3 kcal/mol), followed by H-migration, HF-elimination, and C-C and C-F bond fission. The atmospheric lifetime of C 3 F 4 was estimated to be about 13 days, which is indicative of a very short-lived substance in the atmosphere. Further degradation of the energy-rich C 3 F 4 OH* intermediates by O 2 takes place spontaneously in view of the successive barrier-free and highly exothermic pathways, producing a variety of fluorinated acids, anhydrides, biacetyls, and regenerating OH radicals. For comparison, the reactions of C 3 H 4 , CF 3 CCH, and CH 3 CCF with OH radicals were examined to clarify the F-substitution effect. It is revealed that the reactivity of fluoropropynes could be either reduced by CF 3 or enhanced by atomic F attached to the acetylenic carbon. The present work provides a fundamental understanding of the reactions of fluoroalkynes with OH/O 2 . The use of C 3 F 4 as a promising eco-friendly gaseous dielectric alternative to SF 6 has been supported.

Laryngeal squamous cell carcinoma (LSCC) is a common malignancy of the head and neck. Recently, circular RNA (circRNA) has been studied extensively in multisystem diseases. However, there are few research on biological functions and molecular mechanisms of circRNAs in LSCC. CircRNA array was used to detect the differentially expressed circRNAs. Kaplan-Meier and cox regression analysis were used to identify survival based on circMMP9. The qRT-PCR, RNase R treatment, sanger sequencing and in situ hybridization were used to verify circMMP9 expression, characteristics and localization in LSCC tissues and cells. Functionally, colony formation, MTS, transwell and in vivo assays were proceeded to detect the biological function of circMMP9 in LSCC progression. The RNA-seq was conducted to identify the molecular targets of circMMP9. Mechanically, MeRIP, RNA Immunoprecipitation (RIP), RNA pulldown, Chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were carried on to verify the regulatory mechanism of circMMP9. CircMMP9 was discovered upregulated in LSCC tissues and cells, and high level of circMMP9 was associated with poor prognosis, low degree of pathological grading, high TNM stage and lymph node metastasis of LSCC. CircMMP9 knockdown prevented LSCC progression both in vitro and in vivo, whereas, circMMP9 overexpression had the opposite effect. CircMMP9 was stabilized by IGF2BP2 in m6A-dependent manner. TRIM59 was identified as downstream target of circMMP9. CircMMP9 recruited ETS1 to stimulate TRIM59 transcription. Moreover, TRIM59 accelerated LSCC progression via activating the PI3K/AKT signal pathway. Our findings offered a unique regulatory mechanism for circMMP9 in LSCC, as well as a novel proof that circMMP9 may be utilize as a diagnostic marker and therapeutic target for LSCC patients., (© 2024. The Author(s).)

In situ observations under typhoon conditions are sparse and limited. Distributed acoustic sensing (DAS) is an emerging technology that uses submarine optical-fiber (OF) cables to monitor the sea state. Here, we present DAS-based ocean current observations when a super typhoon passed overhead. The microseismic noise induced by ocean surface gravity waves (OSGWs) during Typhoon Muifa (2022) is observed in the ~0.08-0.38 Hz frequency band, with high-frequency (>0.3 Hz) component being tidally modulated. The OSGW propagation along the entire cable is successfully revealed via frequency-wavenumber analysis. Further, a method based on the current-induced Doppler shifts of DAS-recorded OSGW dispersions is proposed to calculate both speeds and directions of horizontal ocean currents. The measured current is consistent with the tidally induced sea-level fluctuations and sea-surface winds observed at a nearby ocean buoy. These observations demonstrate the feasibility of monitoring the ocean current under typhoon conditions using DAS-instrumented cables., (© 2024. The Author(s).)

Transcription factors with basic helix-loop-helix (bHLH) structures regulate plant growth, epidermal structure development, metabolic processes, and responses to stress extensively. Sea lavender ( Limonium bicolor ) is a recretohalophyte with unique salt glands in the epidermis that make it highly resistant to salt stress, contributing to the improvement of saline lands. However, the features of the bHLH transcription factor family in L. bicolor are largely unknown. Here, we systematically analyzed the characteristics, localization, and phylogenetic relationships of 187 identified bHLH family genes throughout the L. bicolor genome, as well as their cis -regulatory promoter elements, expression patterns, and key roles in salt gland development or salt tolerance by genetic analysis. Nine verified L. bicolor bHLH genes are expressed and the encoded proteins function in the nucleus, among which the proteins encoded by Lb2G14060 and Lb1G07934 also localize to salt glands. Analysis of CRISPR-Cas9-generated knockout mutants and overexpression lines indicated that the protein encoded by Lb1G07934 is involved in the formation of salt glands, salt secretion, and salt resistance, indicating that bHLH genes strongly influence epidermal structure development and stress responses. The current study lays the foundation for further investigation of the effects and functional mechanisms of bHLH genes in L. bicolor and paves the way for selecting salt-tolerance genes that will enhance salt resistance in crops and for the improvement of saline soils., Competing Interests: The authors state that they do not have conflicts of interest related to this work., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nanjing Agricultural University.)

Introduction: T cells are the core of the cellular immunity and play a key role in the regulation of intestinal immune homeostasis. In order to explore the impact Moniezia benedeni ( M. benedeni ) infection on distributions of CD3 + T cells in the small intestine of the sheep., Methods: In this study, sheep pET-28a-CD3 recombinant plasmid were constructed and expressed in BL21 receptor cells, then the rabbit anti-sheep CD3 polyclonal antibody was prepared through recombinant protein inducing. The M. benedeni -infected sheep (infection group, n  = 6) and healthy sheep (control group, n  = 6) were selected, and the distributions of CD3 + T cells in intestinal laminae propria (LP) and mucous epitheliums were observed and analyzed systematically., Results: The results showed that the rabbit anti-sheep CD3 polyclonal antibody had good potency and specificity. In the effector area of small intestine, a large number of CD3 + T cells were mainly diffusely distributed in the intestinal LP as well as in the mucous epitheliums, and the densities of intestinal LP from duodenum to jejunum to ileum were 6.01 cells/10 4  μm 2 , 7.01 cells/10 4  μm 2 and 6.43 cells/10 4  μm 2 , respectively. Their distribution densities in mucous epitheliums were 6.71 cells/10 4  μm 2 , 7.93 cells/10 4  μm 2 and 7.21 cells/10 4  μm 2 , respectively; in the infected group, the distributions of CD3 + T cells were similar to that of the control group, and the densities in each intestinal segment were all significantly increased ( p  < 0.05), meanwhile, the total densities of CD3 + T cells in duodenum, jejunum and ileum were increased by 33.43%, 14.50%, and 34.19%. In LP and mucous epitheliums, it was increased by 33.57% and 27.92% in duodenum; by 25.82% and 7.07% in jejunum, and by 27.07% and 19.23% in ileum, respectively., Discussion: It was suggested that M. benedeni infection did not change the spatial distributions of CD3 + T cells in the small intestine of sheep, but significantly increased their densities, which lays a foundation for further research on the regulatory mechanism of sheep intestinal mucosal immune system against M. benedeni infection., Competing Interests: BX was employed by Lanzhou Safari Park Management Co. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Chai, Yao, Pan, Huang, Wang, Xu, Fan, He, Wang and Zhang.)

As the downstream component of the mitogen-activated protein kinases (MAPK) pathway, the extracellular signal-regulated kinase (ERK) is responsible for phosphorylating a broad range of substrates in cell proliferation, differentiation, and survival. Direct targeting the ERK proteins by the piperidinopyrimidine urea-based inhibitors has been demonstrated to be an effective way to block the MAPK signaling pathway in inhibiting tumor growth. In order to discover better inhibitors, a computer-aided drug design (CADD) approach was employed to reveal the pharmacological characteristics and mechanisms of action. The pharmacophore model was generated on the basis of the compounds with eight features, i.e., four hydrogen bond acceptor atoms, one hydrogen bond donor atom, and three hydrophobic centers. A total of 14 hit compounds were obtained through virtual screening. Two potential inhibitors, namely VS01 and VS02, have been identified by molecular docking and molecular dynamics simulations. Both compounds are capable of attaching to the ERK pocket precisely. The binding free energies of VS01 and VS02 are about 15 kJ/mol and 4 kJ/mol stronger than that of the clinic Ulixertinib because of the characteristic hydrogen bonding, electrostatic, and hydrophilic interactions. The present theoretical investigations shed new light on the rational design of the potential ERK inhibitors to stimulate further experimental tests.Communicated by Ramaswamy H. Sarma.

Salt glands are the unique epidermal structures present in recretohalophytes, plants that actively excrete excess Na + by salt secretory structures to avoid salt damage. Here, we describe a transmembrane protein that localizes to the plasma membrane of the recretohalophyte Limonium bicolor. As virus-induced gene silencing of the corresponding gene LbRSG in L. bicolor decreased the number of salt glands, we named the gene Reduced Salt Gland. We detected LbRSG transcripts in salt glands by in situ hybridization and transient transformation. Overexpression and silencing of LbRSG in L. bicolor pointed to a positive role in salt gland development and salt secretion by interacting with Lb3G16832. Heterologous LbRSG expression in Arabidopsis enhanced salt tolerance during germination and the seedling stage by alleviating NaCl-induced ion stress and osmotic stress after replacing or deleting the (highly) negatively charged region of extramembranous loop. After screened by immunoprecipitation-mass spectrometry and verified using yeast two-hybrid, PGK1 and BGLU18 were proposed to interact with LbRSG to strengthen salt tolerance. Therefore, we identified (highly) negatively charged regions in the extramembrane loop that may play an essential role in salt tolerance, offering hints about LbRSG function and its potential to confer salt resistance., (© 2023 Society for Experimental Biology and John Wiley & Sons Ltd.)

Here, we aimed to optimize the ethanol extraction technology for Yujin powder (YJP) and evaluate its safety. The ultrasonic-assisted ethanol reflux extraction method refluxing was used to extract YJP. The parameters were optimized through a combination of single-factor and response surface methodology (RSM). The comprehensive Y value score calculated using the content of 13 active ingredients in YJP ethanolic extracts (YEEs) and the yield of the dry extract were used as measuring criteria. RSM with a Box-Behnken design using three factors and three levels was adopted to optimize the ethanol extraction technology for YJP. Finally, acute and subchronic toxicity tests were performed to evaluate its safety. The results revealed the best technological parameters: a liquid-material ratio of 24:1, an ethanol concentration of 69%, assistance of ultrasound (40 °C, 50 kHZ, 30 min), reflux time of 53 min, and reflux temperature of 50 °C. In acute toxicity tests, the maximum administration dosage in mice was 28.21 g/kg, which is higher than 10 times the clinical dosage. Adverse effects in the acute and subchronic toxicity tests were not observed. All clinical indexes were normal. In conclusion, the RSM based on AHP-CRITIC weight analysis could be used to optimize the ethanol extraction technology for YJP and YEEs prepared under the above conditions and ensure high safety.

The aim of this study was to investigate the effect of oregano essential oil on IgA + , IgG + , and IgM + cells in the jejunum of castrated Holstein bulls. Twelve castrated Holstein bulls were randomly divided into control (YCK) and oregano essential oil (YEO) groups. Pathological changes in the jejunum were observed by HE staining, and the expression levels of IgA, IgG, and IgM in the jejunum were detected by ELISA. The distributions of IgA + , IgG + , and IgM + cells in the jejunum were analysed by multiplex immunofluorescence and immunohistochemistry. The results showed that the jejunal villi were detached in the YCK group, which may have been related to inflammation, while the intestinal epithelium was clear and intact in the YEO group. The expressions of IgA, IgG, and IgM were significantly reduced by 40.75%, 30.76%, and 50.87%. The IgA + , IgG + , and IgM + cells were diffusely distributed in the lamina propria of the jejunum, and were reduced by 17.07%, 6.44%, and 6.15%, respectively. Oregano essential oil did not alter the distribution characteristics of IgA + , IgG + , or IgM + cells in the jejunum, but it suppressed inflammatory response, decreased immunoglobulin content, and significantly enhanced the formation of an immune barrier in the gastrointestinal mucosa.

In recent years, numerous genes that encode proteins with specific domains that participate in different biological processes or have different molecular functions have been identified. A class of genes with typical domains whose function has rarely been identified and another type of genes with no typical domains have attracted increasing attentions. As many of these so-called as unknown/uncharacterized (U/U) genes are involved in important processes, such as plant growth and plant stress resistance, there is much interest in deciphering their molecular roles. Here, we summarize our current understanding of these genes, including their structures, classifications, and roles in plant growth and stress resistance, summarize progress in the methods used to decipher the roles of these genes, and provide new research perspectives. Unveiling the molecular functions of unknown/uncharacterized genes may suggest strategies to fine-tune important physiological processes in plants, which will enrich the functional network system of plants and provide more possibilities for adaptive improvement of plants., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Wang, Wang and Yuan.)

Hepatic ischemia-reperfusion injury (HIRI) is a complex pathological phenomenon dominated by the innate immune system and involves a variety of immune cells. This condition frequently occurs during hepatectomy, liver transplantation or hemorrhagic shock. HIRI represents an important factor in the poor prognosis of patients after liver surgery. However, there is still a lack of effective intervention to reduce the incidence of HIRI. In this study, we aimed to describe the overall structure of scientific research on HIRI over the past 20 years and provide valuable information and guidelines for future researchers. Bibliometric analysis was used to comprehensively review developments in HIRI and changes in our understanding of HIRI over the past two decades. We identified a total of 4267 articles on HIRI that were published over the past 20 years of which basic research was predominant. Collaboration network analysis revealed that China, the University of California Los Angeles, and Ronald W Busuttil were the most influential country, institute, and scholar, respectively. Co-occurrence cluster analysis revealed that ischemic preconditioning, liver cirrhosis, hepatic I/R injury, autophagy, acute liver failure, oxygen, donation after circulatory death, Nlrp3, remote organ, and microdialysis were the top 10 clusters. Keyword burst detection indicated that autophagy, inflammation, and early allograft dysfunction represent the current research hotspots. In summary, this is the first bibliometric analysis of HIRI research. Our timely analysis of these hotpots and research trends may provide a framework for future researchers and further promote research on the key mechanisms and therapeutic measures in this field., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Background: Circular RNA (circRNA) is a novel noncoding RNA (ncRNA) that plays a critical role in various cancers. However, the clinical significance, biological function, and molecular mechanisms of circRNAs in laryngeal squamous cell carcinoma (LSCC) remain unclear., Methods: A circRNA array was performed to identify the differentially expressed circRNAs. In vitro and in vivo assays were proceeded to verify the biological function of circCDK1 in LSCC. RNA pulldown assays and RNA immunoprecipitation (RIP) were used to confirm the binding between circCDK1 and insulin-like growth factor 2 mRNA binding protein 2(IGF2BP2). The MeRIP assay was then used to identified the N6-methyladenisine (m6A) methylation of calcineurin like phosphatase domain containing1 (CPPED1)., Results: Hsa&#95;circ&#95;0005774 (circCDK1) was found upregulated in LSCC tissues compared to adjacent normal tissues. The level of circCDK1 was positively correlated with poor prognosisof LSCC patients. In vitro and in vivo, circCDK1 promoted migration and invasion of LSCC cells. Mechanistically, eukaryotic translation initiation factor4A3(EIF4A3) induced biogenesis of circCDK1 by binding to its flanking. By competitively binding to IGF2BP2, circCDK1 blocked the m6A modification of CPPED1 in IGF2BP2-dependent manner. Moreover, the circCDK1-mediated decrease of CPPED1 activated the PI3K/AKT signal pathway to facilitate progression of LSCC., Conclusions: Our findings demonstrated that EIF4A3-induced upregulation of circCDK1 promoted LSCC metastasis via EIF4A3-circCDK1-IGF2BP2-CPPED1 to activate PI3K-AKT signal pathway. CircCDK1 might serve as a new diagnostic and prognostic marker or potential therapeutic target for LSCC., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

As an important cancer therapeutic target, extracellular signal-regulated kinases (ERK) are involved in triggering various cellular responses in tumors. Regulation of the ERK signaling pathway by the small molecular inhibitors is highly desired for the sake of cancer therapy. In contrast to the routine inhibitors targeting ERKs through long-range non-bonding interactions, Ponatinib, a covalent inhibitor to ERK2 with a macrocyclic structure characterized by the α,β-C=C unsaturated ketone, can form the stable -C(S)-C(H)-type complex via the four-center barrier due to the nucleophilic addition reaction of the thiol group of the Cys166 residue of ERK2 with the C=C double bond of Ponatinib with reaction free-energy barrier of 47.2 kcal/mol. Reaction mechanisms for the covalent binding were calculated using QM/MM methods and molecular dynamics simulations. The interaction modes and the corresponding binding free energies were obtained for the non-covalent and covalent complexation. The binding free energies of the non-covalent and covalent inhibitions are 14.8 kcal/mol and 33.4 kcal/mol, respectively. The mechanistic study stimulated a rational design on the modified Ponatinib structure by substituting the C=C bond with the C=N bond. It was demonstrated that the new compound exhibits better inhibition activity toward ERK2 in term of both thermodynamic and kinetic aspects through the covalent binding with a lower reaction free-energy barrier of 23.1 kcal/mol. The present theoretical work sheds new light on the development of the covalent inhibitors for the regulation of ERKs.

Background: Metabolic reprogramming is a key marker in the occurrence and development of tumors. This process generates more reactive oxygen species (ROS), promoting the development of oxidative stress. To prevent ROS from harming tumor cells, tumor cells can increase the production of reducing agents to counteract excessive ROS. NMRAL2P has been shown to promote the production of reductive mRNA and plays an important role in the process of oxidative stress., Methods: In this study, the clinical data and RNA sequencing of head and neck tumors were obtained from The Cancer Genome Atlas data set. The long non-coding RNA (LncRNA) related to oxidative stress were then identified using differential and correlation analyses. The differential expression and prognosis of the identified lncRNA were then verified using samples from the library of the Second Hospital of Hebei Medical University. Only NMRAL2P was substantially expressed in cancer tissues and predicted a poor prognosis. The tumor-promoting impact of NMRAL2P was then confirmed using in vitro functional assays. The data set was then split into high- and low-expression subgroups based on the median gene expression of NMRAL2P to obtain the mRNA that had a large difference between the two groups, and examine the mechanism of NMRAL2P on GPX2 using quantitative real-time PCR, RNA binding protein immunoprecipitation assay, and chromatin immunoprecipitation. Mass spectrometry was used to identify NMRAL2P -binding proteins and western blotting was used to investigate probable mechanisms., Results: The lncRNA NMRAL2P is associated with oxidative stress in head and neck tumors. In vitro functional assays showed that the gene has a cancer-promoting effect, increasing lactic acid and superoxide dismutase production, and reducing the production of ROS and malondialdehyde. NMRAL2P promotes the transcription of GPX2 by binding to transcription factor Nrf2. The gene also inhibits the degradation of ENO1, a crucial enzyme in glycolysis, by binding to protein ENO1., Conclusions: This study shows that NMRAL2P can promote glycolysis and reduce the harm to tumor cells caused by ROS. The gene can also be used as a possible target for the treatment of head and neck tumors., Competing Interests: The authors declare there are no competing interests., (©2023 Nie et al.)

Gases for electric insulation are essential for various types of high-voltage power equipment. Sulfur hexafluoride (SF 6 ) has been a dielectric medium commonly used in electrical grids for decades but it is the most potent industrial greenhouse gas. The continuous increase of SF 6 emissions in the atmosphere exerts a significant impact on global warming. The identification of suitable drop-in replacements for all SF 6 -filled apparatuses has been elusive experimentally and theoretically. We claim that tetrafluoropropyne, C 3 F 4 , is a breakthrough in chemical alternatives to SF 6 . The performance of C 3 F 4 was assessed systematically in a 6-dimensional manner, including dielectric strength, liquefaction temperature, global warming potential, thermal stability, toxicity, and arc interruption. On the basis of the extensive ab initio calculations, it has been demonstrated rigorously that C 3 F 4 is an environmentally sustainable solution that may fulfill the complex combination of performance, stability, safety, and environmental properties, namely, the dielectric strength is about 50% higher than that of SF 6 , the boiling point is -50 °C, the GWP for 100 year time horizons is only 3, the decomposition temperature is above 600 °C, the toxicity is as low as HFOs, and the interruption capability is two-thirds of SF 6 . Two protocols are suggested for the practical use of C 3 F 4 . First, equivalence to 0.5 MPa SF 6 could be obtained by filling 0.33 MPa C 3 F 4 pure gas and lead minimum operating temperature down to -21 °C. Second, by taking advantage of synergism effect, the 40% C 3 F 4 /60% CO 2 mixture is a viable alternative to SF 6 with the operating temperature -30 °C without causing any environmental and safety concerns. The present theoretical work sheds new light on the challenging topic of the development of alternative dielectric gases and may stimulate experimental tests on the electrical applications of C 3 F 4 in the future.

The PI3K/Akt signalling pathway is associated with the occurrence and development of tumours and significantly affects the prognosis of patients. We established a predictive signature based on the PI3K/Akt pathway to predict the prognosis of patients. The RNA-seq and clinical data of laryngeal cancer patients were downloaded from The Cancer Genome Atlas (TCGA) database. Three lncRNAs (MNX1-AS1, LINC00330, LSAMP-AS1) were selected through univariate, multivariate Cox and log-rank test analysis to establish a prognostic signature. The patients were then divided into high-risk and low-risk groups based on their risk score. In the TCGA training set, the survival time of the high-risk group was shorter than that of the low-risk group (P < 0.01). Follicular helper T cells were lower in the high-risk group (P = 0.022), and CCR, inflammation promotion, parainflammation, and type I IFN immune function were suppressed. The results of the drug sensitivity analysis suggest that the high-risk group is sensitive to AKT inhibitors. The establishment of the signature was also verified based on the clinical data. Three lncRNAs can facilitate the migration, invasion, and vitality of cancer cells in vitro, and vice versa. Moreover, p-AKT (Ser473) and p-PI3K were highly activated in the cells overexpressing the abovementioned three lncRNAs. The PI3K/Akt signalling pathway-associated prognosis signature has a good predictive effect., (© 2023. Springer Nature Limited.)

Laryngeal squamous cell carcinoma (LSCC) is a common malignant tumor in the head and neck, the 5-year relative survival rate of patients diagnosed with laryngeal cancer was estimated to be 61% from 2012 to 2018. An increasing number of studies have shown that circular RNAs (circRNAs) play a key role in the occurrence and development of cancer and may function as cancer biomarkers and new therapeutic targets. At present, the research on the relationship between circRNAs and LSCC is still in its infancy and needs further exploration. In this study, we found a circRNA (hsa&#95;circ&#95;0001445) associated with LSCC based on bioinformatics analysis. Quantitative real-time polymerase chain reaction (qRT-PCR) assay indicated that the expression of hsa&#95;circ&#95;0001445 was down-regulated in LSCC tissues and cell lines. Notably, the expression of hsa&#95;circ&#95;0001445 was negatively correlated with aggressive clinicopathological features and poor prognosis. Then, functional experiments found that overexpression of hsa&#95;circ&#95;0001445 inhibited the proliferation, migration and invasion of LSCC cells and tumor growth in vivo. Mechanistically, RNA immunoprecipitation (RIP), biotin-labeled probe pull-down, luciferase reporter assay and western blot experiments were employed and found that EIF4A3 reduced the expression of hsa&#95;circ&#95;0001445, and the direct binding of hsa&#95;circ&#95;0001445 to hsa-miR-432-5p attenuated the inhibitory effect of hsa-miR-432-5p on RGMA. In summary, our research suggests that hsa&#95;circ&#95;0001445 may be used as a potential prognostic biomarker and therapeutic target for LSCC.

High hydrostatic pressure (HHP) could induce changes in the structure and properties of starch. Native corn starch was treated and octenyl succinic anhydride (OSA)-modified corn starch was prepared under different pressures (200, 350, 500 and 600 MPa) at 40℃ for 20 min. The mechanism of HHP on the chemical activity of starch was elucidated by analyzing the relationship between the changes of native starch structure and properties and the quality of OSA-modified starch. Results showed that HHP not only helped water and OSA to penetrate the starch granules but also made the structure of starch granules undergone three changes similar to mechanochemical effects. The starch granules treated by 200 MPa were in the stress stage, and the starch granules treated by 500 MPa were in the transition stage from aggregation to agglomeration. Proper pressure treatment could significantly improve chemical activity of starch and quality of OSA-modified starch., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

In view of the significant greenhouse effect of sulfur hexafluoride and the potential biotoxic hazard of perfluorinated substances, we proposed that nitryl cyanide (NCNO 2 ), a nearly nonpolar molecule with a unique combination of two strongly electronegative and polarized functional groups, is a novel fluorine-free replacement to be used as the insulating gas in green electrical grids. Atmospheric chemistry of NCNO 2 has been investigated theoretically to assess its environmental impact if emitted into the atmosphere. Potential energy surfaces for the reaction of NCNO 2 with OH in the presence of O 2 were calculated using the restricted open-shell complete basis set quadratic Becke3 and Gaussian-4 methods on the basis of the density functional (M06-2X) and couple-cluster (CCSD) optimized geometrical parameters. The oxidation of NCNO 2 takes place via the nearly zero-barrier association of OH with the cyano-C to form energy-rich adducts NC(OH)NO 2 , followed by C-N bond rupture to the major HOCN + NO 2 and the minor HONO + NCO products. Interception of the adduct by O 2 can result in OH-regeneration together with further degradation to CO and NO x . Moreover, photolysis of NCNO 2 under tropospheric sunlight conditions might compete with OH-oxidation. The atmospheric lifetime and radiative efficiency of NCNO 2 were computed to be far less than those of either nitriles or nitro compounds. The global warming potential of NCNO 2 was estimated to be in the range of 0-5 for a 100 year time horizon. However, the secondary chemistry of NCNO 2 should be treated with caution in view of the production of NO x in the atmosphere.

Root hairs tie the root system to the soil substrate, facilitate water and nutrient absorption, and enable the interaction with microbes in the soil. Root hair development can be classified into three main development types (I-III). Root hair development type III has been extensively studied, mainly represented using the model plant Arabidopsis thaliana . Transcription factors, plant hormones, and proteins are involved at different root hair developmental stages. The mechanisms underlying development in types I and II have been examined using other representative plant species but have not been studied as intensively. Many key developmental genes in types I and II are highly homologous with those in type III, exhibiting conservation of related mechanisms. Root hairs are also involved in the regulation of plant response to abiotic stress by altering developmental patterns. Abiotic stress, regulatory genes, and plant hormones jointly regulate root hair development and growth; however, few studies have focused on how root hair recognizes abiotic stress signals. This review examines the molecular mechanisms of root hair development and adaptations under stress, and prospective future developments in root hair research are also discussed.

Laryngeal squamous cell carcinoma (LSCC) is a common malignancy among men in the anatomical position of head and neck. Hoarseness, pharyngalgia, and dyspnea are common symptoms. LSCC is a complex polygenic carcinoma that is caused by many factors involving polygenic alteration, environmental pollution, tobacco, and human papillomavirus. Classical protein tyrosine phosphatase nonreceptor type 12 (PTPN12) has been extensively studied to decipher its mechanism as a tumor suppressor gene in various human carcinomas; however, there is no comprehensive elucidation of the PTPN12 expression and its regulatory mechanisms in LSCC. As such, we expect to provide new insights for finding new biomarkers and effective therapeutic targets in LSCC. Immunohistochemical staining, western blot (WB), and quantitative real-time RT-PCR (qRT-PCR) were used for the messenger RNA (mRNA) and protein expression analyses of PTPN12, respectively. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H -tetrazolium, clone formation, transwell migration, and transwell invasion assays were used to assess the proliferation, migration, and invasion ability of LSCC cells. Online prediction and design software tools (http://www.targetscan.org/ and http://www.microRNA.org) were used to predict associated miRNA. Studying the targeted regulatory relationship between miR-146b-3p and PTPN12 was based on dual luciferase reporter gene analysis. qRT-PCR was used to assess miR-146b-3p expression in LSCC. miR-146b-3p inhibitor and mimic were transfected, followed by qRT-PCR and WB assays to detect the expression of PTPN12. The gain and loss functional experiments were used to investigate the effects of miR-146b-3p transfection on the proliferation, migration, and invasion of tumor cells. Online bioinformatics prediction software (https://cn.string-db.org/ and https://www.genecards.org/) was used to determine potential downstream target genes of PTPN12. qRT-PCR and WB analyses were used to assess the mRNA and protein expression levels of target genes. Our study showed significantly decreased mRNA and protein expression levels of PTPN12 in LSCC compared with the adjacent normal tissues. The lower PTPN12 mRNA expression was correlated with pathological differentiation, and lower PTPN12 protein expression was correlated with the TNM stage in LSCC tissues. The subsequent in vitro functional analyses showed the inhibitory effect of PTPN12 over-expression on the proliferation, migration, and invasiveness abilities of LSCC cell line. Using online prediction and design software, miR-146b-3p was searched to target PTPN12. The miR-146b-3p was expressed at a high level in LSCC tissues and cell lines. Luciferase reporter assay exhibited that miR-146b-3p inhibited the luciferase activity of PTPN12 markedly. The functional analyses showed the tumor-promoting role of miR-146b-3p on the proliferation, migration, and invasiveness abilities of LSCC cell. Furthermore, co-transfection of cells with miR-146b-3p and PTPN12 significantly restored the inhibitory effect of PTPN12 on LSCC cell growth, migration, and invasiveness. This phenomenon unveiled that miR-146b-3p regulated the proliferation, migration, and invasion of LSCC cells by targeting PTPN12. EGFR and ERBB2 were selected as the downstream-regulation target genes. Up-regulation of PTPN12 significantly suppressed EGFR expression. Accordingly, the miR-146b-3p mimic significantly up-regulated the EGFR expression. However, up-regulation of PTPN12 and miR-146b-3p mimic suppressed ERBB2 protein expression but induced its gene expression. Down-regulation of PTPN12 is associated with up-regulation of miR-146b-3p in LSCC. Moreover, PTPN12 serves as a tumor suppressor gene through regulating the proliferation, migration, and invasion of LSCC cells. miR-146b-3p/PTPN12 axis is expected to be a novel therapeutic target in LSCC., Competing Interests: Conflict of interest: The authors declare no conflict of interest., (© 2023 the author(s), published by De Gruyter.)

Halophytes complete their life cycles in saline environments. The recretohalophyte Limonium bicolor has evolved a specialized salt secretory structure, the salt gland, which excretes Na + to avoid salt damage. Typical L. bicolor salt glands consist of 16 cells with four fluorescent foci and four secretory pores. Here, we describe a special type of salt gland at the base of the L. bicolor leaf petiole named bracelet salt glands due to their beaded-bracelet-like shape of blue auto-fluorescence. Bracelet salt glands contain more than 16 cells and more than four secretory pores. Leaf disc secretion measurements and non-invasive micro-test techniques indicated that bracelet salt glands secrete more salt than normal salt glands, which helps maintain low Na + levels at the leaf blade to protect the leaf. Cytokinin treatment induced bracelet salt gland differentiation, and the developed ones showed no further differentiation when traced with a living fluorescence microscopy imager, even though new salt gland development and leaf expansion were observed. Transcriptome revealed a NAC transcription factor gene that participates in bracelet salt gland development, as confirmed by its genome editing and overexpression in L. bicolor. These findings shed light on bracelet salt gland development and may facilitate the engineering of salt-tolerant crops., (© 2022 The Authors. Journal of Integrative Plant Biology published by John Wiley & Sons Australia, Ltd on behalf of Institute of Botany, Chinese Academy of Sciences.)

Long non-coding RNAs (lncRNAs) are RNAs of more than 200 nucleotides in length that are not (or very rarely) translated into proteins. In eukaryotes, lncRNAs regulate gene expression at the transcriptional, post-transcriptional, and epigenetic levels. lncRNAs are categorized according to their genomic position and molecular mechanism. This review summarized the characteristics and mechanisms of plant lncRNAs involved in vegetative growth, reproduction, and stress responses. Our discussion and model provide a theoretical basis for further studies of lncRNAs in plant breeding., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Wang, Fan, Wang and Yuan.)

As a potential insulation gas to replace sulfur hexafluoride (SF 6 ) due to environmental concerns, trifluoromethanesulonyl fluoride (CF 3 SO 2 F) has attracted great interests in various high-voltage electric applications. Thermal stability of CF 3 SO 2 F plays an important role in the rational design of the gas-insulated electric equipment. Unimolecular decomposition of CF 3 SO 2 F was investigated using high-level ab initio methods including the explicitly correlated RCCSD(T)-F12, the composite ROCBS-QB3, and the multireference RS2 extrapolated to complete basis set limit on the basis of M06-2X-, B2PLYPD3-, and CCSD-optimized geometrical parameters. Rate coefficients and decomposition temperatures were simulated using master equations. CF 3 SO 2 F decomposes predominantly via a simple C-S bond cleavage to form CF 3 and SO 2 F, accompanied by a roaming induced F-abstraction detour to release CF 4 and SO 2 , or isomerizes via CF 3 migration to the more stable CF 3 OSFO followed by the production of CF 2 O and SOF 2 . Various characteristic decomposition products (e.g., CF 4 , C 2 F 6 , CF 2 O, SO 2 , SOF 2 , SO 2 F 2 , CF 3 H, and so on.) have been identified theoretically through secondary reactions and hydrolysis of CF 3 SO 2 F in the presence of water vapor. Electronic structures and stability of CF 3 SO 2 F could be affected significantly by the external electric field orientated along the S-C bond. The field-dependent electron-molecule capture rates support that CF 3 SO 2 F is superior to SF 6 in dielectric strength. The present computational findings shed light on the practical use of CF 3 SO 2 F as the replacement gas for SF 6 .

Sweet sorghum [Sorghum bicolor (L.) Moench], a C4 crop with high biomass and strong resistance to multiple stresses, can grow and reproduce in saline-alkaline soil and is an ideal raw material for biofuels. Under high-salinity conditions, sweet sorghum shows extensive salt exclusion. However, the specific molecular mechanism of the apoplastic barrier in salt exclusion is unknown. In this study, SbCASP-LP1C1 (a CASP-like protein1C1) was localized in the plasma membrane of sweet sorghum root endodermal cells, and its function was further studied by heterologous expression in Arabidopsis (35 S:SbCASP-LP1C1-GFP). When germinated and grown on 50 mM NaCl, the SbCASP-LP1C1-expressing lines had longer roots and a higher salinity threshold compared with wild-type (Col-0) plant and the casp-lp T-DNA insertion mutant in Arabidopsis. The 35 S:SbCASP-LP1C1-GFP lines also suffered less oxidative damage as determined by DAB and NBT staining, and the expression levels of several antioxidant genes were higher in these lines. Moreover, the stele of 35 S:SbCASP-LP1C1-GFP lines was less permeable to propidium iodide, and these plants contained less Na + in their shoots and roots compared to wild type and casp-lp. In the 35 S:SbCASP-LP1C1-GFP lines, the expression levels of two Casparian strip synthesis genes, MYB36 and ESB1, were increased. These results indicate that SbCASP-LP1C1 may be involved in the polymerization of lignin monomers in the Casparian strip of sweet sorghum, thereby regulating salt tolerance. These results provide a theoretical basis to understand the role of plant roots in salt exclusion and a means by which to improve the salt tolerance of crops., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Long noncoding RNAs (lncRNAs) have an effect on the occurrence and progression of a considerable number of diseases, especially cancer. Existing research has suggested that MAGI2 antisense RNA 3 (MAGI2-AS3) takes on a critical significance in the development of hepatocellular carcinoma and lung cancer. However, the functions of MAGI2-AS3 in laryngeal squamous cell carcinoma (LSCC) remain unclear. In this study, MAGI2-AS3 expression level in LSCC tissue and cell lines was detected, and the effect of MAGI2-AS3 overexpressed on LSCC phenotypes and the possible influence mechanisms were examined. MAGI2-AS3 was downregulated in the tissues of LSCC patients versus non-tumor tissues, and it was correlated with advanced TNM (tumor, node, metastasis) stage and lymph node metastases, as indicated by the results of this study. MAGI2-AS3 inhibited the proliferation, migration, and invasion of LSCC cells in vitro and in vivo . Furthermore, the hypermethylation level of the MAGI2-AS3 promoter region was indicated by bisulfite genomic sequencing and methylation-specific polymerase chain reaction, such that MAGI2-AS3 expression was downregulated. Besides, MAGI2-AS3 promoter hypermethylation was regulated by DNA methyltransferase 1 (DNMT1), and MAGI2-AS3 expression was reversed by 5-Aza-2'-deoxycytidine (5-Aza). Moreover, the result of the RNA pull-down experiment suggested that 38 proteins were enriched in the MAGI2-AS3 group versus the control group in TU177 cells. To be specific, SPT6 (ie, a conserved protein) was enriched by fold change >10. SPT6 knockdown reduced the antitumor effect of MAGI2-AS3 in TU177 and AMC-HN-8 cells. Meanwhile, SPT6 overexpression inhibited the proliferation, metastasis, and invasion of TU177 and AMC-HN-8 cells. As revealed by the above findings, DNMT1-regulated MAGI2-AS3 promoter hypermethylation led to downregulated MAGI2-AS3 expression, such that the presence and progression of LSCC were inhibited in an SPT6 binding-dependent manner.

Halophytes can grow and reproduce in high-salinity environments, making them an important reservoir of genes conferring salt tolerance. With the expansion of saline soils worldwide, exploring the mechanisms of salt tolerance in halophytes and improving the salt tolerance of crops have become increasingly urgent. Limonium bicolor is a halophyte with salt glands that secrete excess Na + through leaves. Here, we identified an uncharacterized gene Lb1G04794 , which showed increased expression after NaCl treatment and was high during salt gland development in L. bicolor . Overexpression of Lb1G04794 in L. bicolor showed promoted salt gland development, indicating that this gene may promote salt gland differentiation. Transgenic Arabidopsis strains overexpressing Lb1G04794 showed increased trichomes and decreased root hairs under normal conditions. Compared with wild type (WT), root growth in the transgenic lines was less inhibited by NaCl treatment. Transgenic seedlings accumulated less fresh/dry weight reductions under long-term salt treatment, accompanied by lower Na + and malondialdehyde accumulation than WT, indicating that these transgenic lines behave better growth and undergo less cellular damage under NaCl stress. These results were consistent with the low expression levels of salt-tolerance marker genes in the transgenic lines upon salt stress. We conclude that the unknown gene Lb1G04794 positively regulated salt gland development, and promoted salt tolerance of Arabidopsis, offering a new direction for improving salt tolerance of non-halophytes and crops., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Jiao, Zhao, Wang and Yuan.)

In the context of plant breeding, bioinformatics can empower genetic and genomic selection to determine the optimal combination of genotypes that will produce a desired phenotype and help expedite the isolation of these new varieties. Bioinformatics is also instrumental in collecting and processing plant phenotypes, which facilitates plant breeding. Robots that use automated and digital technologies to collect and analyze different types of information to monitor the environment in which plants grow, analyze the environmental stresses they face, and promptly optimize suboptimal and adverse growth conditions accordingly, have helped plant research and saved human resources. In this paper, we describe the use of various bioinformatics databases and algorithms and explore their potential applications in plant breeding and for research on plant disease resistance.

Soil salinization is one of the major factors restricting crop growth and agricultural production worldwide. Recretohalophytes have developed unique epidermal structures in their aboveground tissues, such as salt glands or salt bladders, to secrete excess salt out of the plant body as a protective mechanism from ion damage. Three hypotheses were proposed to explain how salt glands secrete salts: the osmotic hypothesis, a hypothesis similar to animal fluid transport, and vesicle-mediated exocytosis. However, there is no direct evidence to show whether the salt gland-secreted liquid contains landmark proteins or peptides which would elucidate the salt secretion mechanism. In this study, we collected the secreted liquid of salt glands from Limonium bicolor , followed by extraction and identification of its constituent proteins and peptides by SDS-PAGE and mass spectrometry. We detected 214 proteins and 440 polypeptides in the salt gland-secreted droplets of plants grown under control conditions. Unexpectedly, the proportion of energy metabolism-related proteins increased significantly though only 16 proteins and 35 polypeptides in the droplets of salt-treated plants were detected. In addition, vesicle transport proteins such as the Golgi marker enzyme glycosyltransferase were present in the secreted sap of salt glands from both control and salt-treated plants. These results suggest that trans-Golgi network-mediated vesicular transport and energy production contributes to salt secretion in salt glands.

Transmembrane proteins participate in various physiological activities in plants, including signal transduction, substance transport, and energy conversion. Although more than 20% of gene products are predicted to be transmembrane proteins in the genome era, due to the complexity of transmembrane domains they are difficult to reliably identify in the predicted protein, and they may have different overall three-dimensional structures. Therefore, it is challenging to study their biological function. In this review, we describe the typical structures of transmembrane proteins and their roles in plant growth, development, and stress responses. We propose a model illustrating the roles of transmembrane proteins during plant growth and response to various stresses, which will provide important references for crop breeding.

Background: Age-related hearing loss is an increasing sensorineural hearing loss. But the pathogenesis of ARHL has not been clarified. Herein, we studied the role and significance of PIN1 in regulating autophagy activity in senescence HEI-OC1cells and HCs., Methods and Results: C57BL/6 mice and HEI-OC1 cells were contained in our research. Transfection of plasmids and juglone were used to upregulate or inhibit the PIN 1 expression. Immunofluorescence and Western blot were used to detect the expression of PIN1, LC3, p62, p21 and p16 protein levels in the hair cells of C57BL/6 mice cochleae and HEI-OC1 cells. Senescence-associated β -galactosidase (SA- β -gal) staining was used to investigate the senescent level.The results of this study showed that the level of autophagy increased in the senescent auditory hair cells. When inhibited the autophagy level with 3-MA, the senescent HEI-OC1 cells were alleviated. The autophagy activity in senescent HEI-OC1 cells also could be reduced by overexpressing PIN1 protein. On the contrary, inhibiting PIN1 could increase the autophagy level of senescent cells and cochlear hair cells., Conclusion: PIN1 might regulate autophagy activity to induce the senescent of HEI-OC1cells and HCs, which will provide a theoretical support for the prevention and treatment of age-related hearing loss., Competing Interests: The authors declare there are no competing interests., (©2022 Lv et al.)

Limonium bicolor is a dicotyledonous recretohalophyte with several multicellular salt glands on the leaves. The plant can directly secrete excess salt onto the leaf surface through the salt glands to maintain ion homeostasis under salt stress. Therefore, it is of great significance to study the functions of genes related to salt gland development and salt tolerance. In this study, an R1-type MYB transcription factor gene was screened from L. bicolor , named LbMYB48 , and its expression was strongly induced by salt stress. Subcellular localization analysis showed that LbMYB48 was localized in the nucleus. LbMYB48 protein has transcriptional activation activity shown by transcriptional activation experiments. The density of salt glands in the leaves and the salt secretion capacity of LbMYB48 -silenced lines were decremented, as demonstrated by the leaf disc method to detect sodium ion secretion. Furthermore, salt stress index experiments revealed that the ability of LbMYB48 -silenced lines to resist salt stress was significantly reduced. LbMYB48 regulates salt gland development and salt tolerance in L. bicolor mainly by regulating the expression of epidermal cell development related genes such as LbCPC-like and LbDIS3 and salt stress-related genes ( LbSOSs , LbRLKs , and LbGSTs ) as demonstrated by RNA-seq analysis of LbMYB48-silenced lines. The heterologous over-expression of LbMYB48 in Arabidopsis thaliana improves salt tolerance of plants by stabilizing ion and osmotic balance and is likely to be involved in the abscisic acid signaling pathway. Therefore, LbMYB48 , a transcriptional activator regulates the salt gland development of L. bicolor and salt tolerance of L. bicolor and A. thaliana., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Han, Qiao, Li, Yang, Zhang, Zhang, Guo, Liu, Wang and Wang.)