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Abstract Background The cellular origin of hypopharyngeal diseases is crucial for further diagnosis and treatment, and the microenvironment in tissues may also be associated with specific cell types at the same time. Normal adjacent tissues (NATs) of hypopharyngeal carcinoma differ from non-tumor-bearing tissues, and can influenced by the tumor. However, the heterogeneity in kinds of disease samples remains little known, and the transcriptomic profile about biological information associated with disease occurrence and clinical outcome contained in it has yet to be fully evaluated. For these reasons, we should quickly investigate the taxonomic and transcriptomic information of NATs in human hypopharynx. Results Single-cell suspensions of normal adjacent tissues (NATs) of hypopharyngeal carcinoma were obtained and single-cell RNA sequencing (scRNA-seq) was performed. We present scRNA-seq data from 39,315 high-quality cells in the hypopharyngeal from five human donors, nine clusters of normal adjacent human hypopharyngeal cells were presented, including epithelial cells, endothelial cells (ECs), mononuclear phagocyte system cells (MPs), fibroblasts, T cells, plasma cells, B cells, mural cells and mast cells. Nonimmune components in the microenvironment, including epithelial cells, endothelial cells, fibroblasts and the subpopulations of them were performed. Conclusions Our data provide a solid basis for the study of single-cell landscape in human normal adjacent hypopharyngeal tissues biology and related diseases.

Abstract Background Endodontic treatment has benefited from the development of new techniques and equipment. Few clinical studies have been published on the complications associated with root canal preparations performed by doctors with different working experiences using contemporary techniques. This study aimed to analyze the complications of endodontic treatment performed by residents and endodontic specialists in a teaching stomatology hospital using contemporary techniques. Methods Cases of root canal treatment (RCT) and non-surgical root canal retreatment (ReRCT) performed by residents with 1–3 years of experience and endodontic specialists with 5–7 years of experience were collected from the electronic medical system of the Department of Endodontics, Beijing Stomatology Hospital, from September 1, 2020 to August 31, 2021. The cases were examined in terms of patient age, sex, type of tooth, diagnosis, treatment modality (RCT or ReRCT), number of appointments, whether an operating microscope was used, presence of ledges, canal transportation, perforations, missed canals, separated instruments, flare-ups and clinical incidence of second mesiobuccal (MB2) root canal in the maxillary molars. Results In total, 859 teeth from 820 patients were included in the analysis. The overall incidence of complications in the resident group was significantly higher than that in the specialist group. More ledges and flare-ups were observed in the resident group (p

Chitosan-pectin gel beads (CPB) have high potential for removing heavy metals from food. This study aimed to improve their stability, recyclability, and adsorption capacity by modified CPB with gelatin (Gel) and carboxymethyl cellulose sodium (CMC). The structural characteristics of the modified CPBs were characterized using Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TG), Zeta potential, scanning electron microscopy (SEM), pore size distribution analysis (BET), X-ray photoelectron spectroscopy (XPS), specific surface area analysis. The adsorption-resolution conditions of modified CPB were optimized, and their actual removal efficiency for Pb(II) in C-phycocyanin was evaluated. Results showed that CMC-modified CPB (CMC-CPB) had higher thermal stability, rougher and more porous surface, larger specific surface area (20.28±1.35 m2/g), lower zeta potential, stronger metal ion adsorption capacity, and higher regeneration efficiency compared with CPB and Gel-CPB. FTIR showed the functional group of CPB had significant difference after modification, and the main group in CPB were carboxyl, hydroxyl, and amino groups. TG analysis presented the thermal stability of CMC-CPB was higher than that of CPB and Gel-CPB. XPS analysis showed CMC-CPB had the strongest absorption peak for Pb(II). The optimal pH and temperature for the three adsorbents (CPB, Gel-CPB, and CMC-CPB) to remove Pb(II) were 6.0 and 60 ℃, respectively. The Pb(II) adsorption process of all three adsorbents fit the Langmuir isotherm model (R2=0.9543~0.9811) and the pseudo-second-order kinetic model (R2=0.9963~0.9991), and the adsorption process belonged to the monolayer chemical adsorption, involving the complexation of -COO, -OH, -CO-NH, and Pb(II). Based on the Langmuir model curve, the maximum adsorption capacity (qmax) of CMC-CPB for Pb(II) was 69.37 mg/g, significantly higher than that of Gel-CPB and CPB (P

During the process of chip formation, the chip is subjected to extrusion pressure, friction, heat, and a strong chemical reaction. The chip’s macro and micro morphology, to a certain extent, reflect the condition of the tool during the cutting procedure. Therefore, researching the macroscopic and microscopic morphology of the chip’s surface in response to different tool wear conditions is of great significance to reproducing the cutting condition and analyzing the tool wear mechanism. This paper focuses on the chips formed by milling the difficult-to-machine material 508III high-strength steel. Firstly, the 508III steel milling experiment is carried out at the actual machining site to collect chip data under different tool wear conditions. Next, the free surface morphology of chips and the bottom surface morphology of chips are analyzed. Further, the chip edges are investigated, and their causes are analyzed. Finally, heavy milling 508III steel chip curl morphology analysis is performed. The research results play important roles in revealing the mechanism of tool wear and the relationship between chip morphology and tool wear. This information can be used to provide theoretical and technical support for monitoring the tool wear status based on chip morphology.

Abstract Head neck squamous cell carcinoma (HNSCC) is one of the most common malignant tumors which ranks the sixth incidence in the world. Although treatments for HNSCC have improved significantly in recent years, its recurrence rate and mortality rate remain high. Myosin genes have been studied in a variety of tumors, however its role in HNSCC has not been elucidated. GSE58911 and GSE30784 gene expression profile analysis were performed to detect significantly dys-regulated myosin genes in HNSCC. The Cancer Genome Atlas (TCGA) HNSCC database was used to verify the dys-regulated myosin genes and study the relationship between these genes and prognosis in HNSCC. The results showed that MYL1, MYL2, MYL3, MYH2, and MYH7 were down-regulated, while MYH10 was up-regulated in patients with HNSCC. Interestingly, MYL1, MYL2, MYH1, MYH2, and MYH7 were shown to be unfavorable prognostic markers in HNSCC. It is also worth noting that MYL1 was a specific unfavorable prognostic biomarker in HNSCC. MYL1, MYL2, MYL3, MYH2, MYH7, and MYH10 promoted CD4 + T cells activation in HNSCC. MYL1 was proved to be down-regulated in HNSCC tissues compared to normal tissues at protein levels. MYL1 overexpression had no effect on proliferation, but significantly promoted migration of Fadu cells. MYL1 increased EGF and EGFR protein expression levels. Moreover, there is a positive correlation between MYL1 expression and Tcm CD8 cells, Tcm CD4 + cells, NK cells, Mast cells, NKT cells, Tfh cells and Treg cells in HNSCC. Overall, MYL1 facilitates tumor metastasis and correlates with tumor immune infiltration in HNSCC and these effects may be associated with the EGF/EGFR pathway.

Objective: To evaluate the dynamic expression profiling alterations of SARS-CoV-2-associated molecules within the fertile human endometrium throughout the menstrual cycle. Furthermore, to explore the inherent vulnerability of the endometrium to SARS-CoV-2 infection among women experiencing recurrent pregnancy failure, including both recurrent implantation failures (RIF) and recurrent pregnancy losses (RPL).Method: The present study employed multiple datasets to investigate the expression patterns of SARS-CoV-2-associated genes. Firstly, a single-cell RNA-sequencing dataset comprising endometrial samples from 19 healthy women across the menstrual cycle was utilized. Additionally, two microarray datasets encompassing 24 women with RIF, and 24 women with RPL during the peri-implantation phase were included. To complement these analyses, immunohistochemical (IHC) staining was performed on endometrial samples collected from 30 women with RIF, 30 women with RPL, and 20 fertile controls recruited specifically during the implantation period.Results: The investigation revealed a moderate expression percentage of CTSL (22%), TMPRSS4 (15%), FURIN (16%) and MX1 (9%) in endometrium. Conversely, the expression percentages of ACE2 (1%) and TMPRSS2 (4%) were relatively low. Notably, the expression of BSG exhibited an increment towards the window of implantation, reaching its peak during the middle secretary phase. Furthermore, a significant reduction (p < 0.05) in TMPRSS2 expression was observed in the RIF group compared to the control group. While the expression of BSG was significantly increased (p < 0.05) in the RPL group, findings that were corroborated by the IHC staining results.Conclusion: The findings of this study indicate a noteworthy upregulation of BSG expression in the endometrium of women with RPL. These results suggest an augmented susceptibility of endometrium to SARS-CoV-2 infection, potentially contributing to unfavorable pregnancy outcomes.

Herein, the effect of current on the solidification microstructure and properties of 35CrMo structural steel has been studied. The effect of an electric field on the solidification structure of an ingot was investigated by immersing two parallel electrodes into the free surface of molten steel. Using the interaction between the current and melt as well as the Lorentz force generated by its own induced magnetic field, the whole region of the melt was covered with an eddy current. The numerical simulation of the ingot solidification process has been carried out and its influence on the inner flow field during the ingot solidification control process discussed. The results showed that an applied electric field caused turbulence inside the ingot, which drove the molten alloys to rotate and stir, refined the solidification structure, reduced the solidification defects, such as shrinkage cavity and segregation, and increased from 549.9 MPa at the top edge of the ingot and 411.4 MPa at the middle edge to 560.2 and 510.2 MPa, respectively. In addition, the electric field made the hardness and strength of each part of the ingot more uniform and improved the quality of its rigidity for the steel production process.

In order to solve the problems of poor hole-filling performance and the high seed-breakage rate of conventional rice bud seed precision hole direct seed-metering devices, a side-filled rice precision hole direct seed-metering device was developed, and the mechanism and force analyses for seeding operations were carried out. The key factors affecting seeding quality were determined: rotation speed, seeding angle and seeding height. By coupling the discrete element method (DEM) and multi-rigid body dynamics (MBD), the seed breakage rate and seeding performance at different rotation speeds were analyzed. Single-factor bench testing was used to analyze the effect of a duckbill unit on seeding performance under different factor levels. The three-factor and five-level quadratic regression orthogonal rotation center combination test methods were used to obtain the optimal working parameter combination. The test results showed that when the rotation speed was 47 r/min, the seeding angle was 19°, and the seeding height was 180 mm, the qualified index of seeding was 92.03%, the hole diameter qualified index was 91.62%, and the hole distance variation index was 7.17%. This study provides a reference for the research of mechanical rice sprouting seed-metering devices.

BackgroundHuman hypopharygeal squamous cell carcinoma (HSCC) is a common head and neck cancer with a poor prognosis in advanced stages. The occurrence and development of tumor is the result of mutual influence and co-evolution between tumor cells and tumor microenvironment (TME). Tumor immune microenvironment (TIME) refers to the immune microenvironment surrounding tumor cells. Studying TIME in HSCC could provide new targets and therapeutic strategies for HSCC.MethodsWe performed single-cell RNA sequencing (scRNA-seq) and analysis of hypopharyngeal carcinoma, paracancerous, and lymphoid tissues from five HSCC patients. Subdivide of B cells, T cells, macrophages cells, and monocytes and their distribution in three kinds of tissues as well as marker genes were analyzed. Different genes of IGHG1 plasma cells and SPP1+ macrophages between HSCC tissues, adjacent normal tissues and lymphatic tissues were analyzed. Additionally, we studied proliferating lymphocytes, T cells exhaustion, and T cell receptor (TCR) repertoire in three kinds of tissues.ResultsTranscriptome profiles of 132,869 single cells were obtained and grouped into seven cell clusters, including epithelial cells, lymphocytes, mononuclear phagocytics system (MPs), fibroblasts, endothelial cells (ECs), plasmacytoid dendritic cells (pDCs), and mast cells. Tumor metastasis occurred in three lymphoid tissues. Four distinct populations were identified from lymphocytes, including B cells, plasma cells, T cells and proliferating lymphocytes. We found IGHA1 and IGHG1 specific plasma cells significantly overexpressed in HSCC tissues compared with normal hypopharygeal tissues and lymphatic tissues. Five distinct populations from MPs were identified, including macrophages, monocytes, mature dendritic cells (DCs), Type 1 conventional dendritic cells (cDC1) and Type 2 conventional dendritic cells (cDC2). SPP1+ macrophages were significantly overexpressed in HSCC tissues and lymphatic tissues compared with normal hypopharygeal tissues, which are thought to be M2-type macrophages. Exhaustion of CD8+ Teff cells occurred in HSCC tissues. At last, we verified that IgA and IgG1 protein expression levels were significantly up-regulated in HSCC tissues compared to adjacent normal tissues.ConclusionOverall, this study revealed TIME in HSCC and lymphatic metastasis, and provided potential therapeutic targets for HSCC.

Abstract Background Camellia oleifera (C. oleifera) is a woody edible oil crop of great economic importance. Because of the lack of modern biotechnology research, C. oleifera faces huge challenges in both breeding and basic research. The protoplast and transient transformation system plays an important role in biological breeding, plant regeneration and somatic cell fusion. The objective of this present study was to develop a highly efficient protocol for isolating and purifying mesophyll protoplasts and transient transformation of C. oleifera. Several critical factors for mesophyll protoplast isolation from C. oleifera, including starting material (leaf age), pretreatment, enzymatic treatment (type of enzyme, concentration and digestion time), osmotic pressure and purification were optimized. Then the factors affecting the transient transformation rate of mesophyll protoplasts such as PEG molecular weights, PEG4000 concentration, plasmid concentration and incubation time were explored. Results The in vitro grown seedlings of C. oleifera ‘Huashuo’ were treated in the dark for 24 h, then the 1st to 2nd true leaves were picked and vacuumed at − 0.07 MPa for 20 min. The maximum yield (3.5 × 107/g·FW) and viability (90.9%) of protoplast were reached when the 1st to 2nd true leaves were digested in the enzymatic solution containing1.5% (w/v) Cellulase R-10, 0.5% (w/v) Macerozyme R-10 and 0.25% (w/v) Snailase and 0.4 M mannitol for 10 h. Moreover, the protoplast isolation method was also applicable to the other two cultivars, the protoplast yield for ‘TXP14’ and ‘DP47’ was 1.1 × 107/g·FW and 2.6 × 107/g·FW, the protoplast viability for ‘TXP14’ and ‘DP47’ was 90.0% and 88.2%. The purification effect was the best when using W buffer as a cleaning agent by centrifugal precipitation. The maximum transfection efficiency (70.6%) was obtained with the incubation of the protoplasts with 15 µg plasmid and 40% PEG4000 for 20 min. Conclusion In summary, a simple and efficient system for isolation and transient transformation of C. oleifera mesophyll protoplast is proposed, which is of great significance in various aspects of C. oleifera research, including the study of somatic cell fusion, genome editing, protein function, signal transduction, transcriptional regulation and multi-omics analyses.

Abstract Pelvic injuries refer to the disruption of the inherent structural and mechanical integrity of the pelvic ring. Sacroiliac screw fixation technique is often applied for the treatment of posterior pelvic ring injury, which is prone to the iatrogenic injury. This study will compare the intraoperative and postoperative variables of patients underwent sacroiliac screw fixation with intraoperative CT and conventional imaging to evaluate the effect. Thirty-two patients with posterior pelvic ring injury treated by sacroiliac screw fixation from January 2019 to December 2020 were enrolled in this retrospective study. All patients were divided into two groups according to the different assistance of intraoperative imaging. Subsequently, the correlative data were compared and analysed statistically. Twelves cases were assigned to intraoperative CT group, and the remaining twenty cases were assigned to conventional group. There was no significant difference in duration of surgery, fracture healing time, time to ambulation, complications, and Matta radiological score. There was statistically significant difference (p

The utilization of primary school buildings is multifaceted, primarily due to the high occupancy density, varying thermal preferences among occupants, diverse indoor activities (such as walking, sports, and conversation), and a constant flow of individuals entering and exiting the building. This results in the frequent opening and closing of external windows and doors and fluctuations in internal heat gain. Consequently, frequent interactions between the indoor and outdoor microenvironments lead to energy losses. This study conducts a comprehensive literature review on building energy loss stemming from occupant behavior and the interactions between indoor and outdoor microenvironments. Furthermore, it proposes a dynamic real-time monitoring system based on a foundation of computer data capture and a visualization platform for building energy loss. The research methods include data crawling, data association rule mining, and data association analysis. The research findings yield a universally applicable and informative building energy-saving design system based on extensive data analysis. Additionally, the system presents information on occupants’ behavior and the microclimate data of indoor and outdoor environments on a computer screen, facilitating human–machine communication and enabling timely adjustments to be made, thus facilitating the construction of design strategies for new buildings and operation and maintenance strategies for existing buildings.

Abstract Background Between 2012 and 2020, the obesity rate increased among Chinese people aged 18 years and above, along with blood pressure, fasting blood glucose, serum total cholesterol, and triglycerides. Purpose Our objective was to compare the effects of a combined intervention, including individualized exercise prescription plus remote management versus individualized exercise prescription only, on cardiovascular risk factors in patients who are overweight or obese, with the aim of establishing a more effective remote model of health management than self-management. Methods This nonrandomized controlled trial (ChiCTR2100046307) studied patients who are overweight or obese at model labor health management centers from January 2019 to December 2019, including 55 people in the experimental group and 34 in the control group. The relevant indexes of all the research objects from both experimental group and control group were examined. Participants in the experimental group were given individualized exercise prescription combined with remote exercise management over a period of 3 months. The control group was prescribed exercise only at time of enrollment and taught about exercise once, followed by voluntary exercise and self-management for 3 months. Result After adjusting for baseline differences, the changes in weight (−2.72 ± 4.03 kg versus 0.32 ± 2.50 kg, P

The wild species field cress (Lepidium campestre) has the potential to become a novel cover and oilseed crop for the Nordic climate. Its seed oil is however currently unsuitable for most food, feed, and industrial applications, due to the high contents of polyunsaturated fatty acids (PUFAs) and erucic acid (C22:1). As the biosynthesis of these undesirable fatty acids is controlled by a few well-known major dominant genes, knockout of these genes using CRISPR/Cas9 would thus be more effective in improving the seed oil quality. In order to increase the level of the desirable oleic acid (C18:1), and reduce the contents of PUFAs and C22:1, we targeted three important genes FATTY ACID ELONGASE1 (FAE1), FATTY ACID DESATURASE2 (FAD2), and REDUCED OLEATE DESATURASE1 (ROD1) using a protoplast-based CRISPR/Cas9 gene knockout system. By knocking out FAE1, we obtained a mutated line with almost no C22:1, but an increase in C18:1 to 30% compared with 13% in the wild type. Knocking out ROD1 resulted in an increase of C18:1 to 23%, and a moderate, but significant, reduction of PUFAs. Knockout of FAD2, in combination with heterozygous FAE1fae1 genotype, resulted in mutated lines with up to 66% C18:1, very low contents of PUFAs, and a significant reduction of C22:1. Our results clearly show the potential of CRISPR/Cas9 for rapid trait improvement of field cress which would speed up its domestication process. The mutated lines produced in this study can be used for further breeding to develop field cress into a viable crop.

Abstract Background Polycystic ovary syndrome (PCOS) is a common endocrine disorder among reproductive-age women. The mechanism by which circular RNA (circRNA) drives PCOS development remains unclear. Thus, the study is designed to explore the role of a novel circRNA, circ_FURIN, in the PCOS cell model and the underlying mechanism. Methods PCOS cell model was established by treating human ovarian granulosa-like tumor cells (KGN) with Testosterone (TTR). RNA expressions of circ_FURIN, microRNA-423-5p (miR-423-5p) and myotubularin 1 (MTM1) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was checked by Western blot. Cell proliferation was investigated by a 5-Ethynyl-29-deoxyuridine assay, 3-(4,5-Dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis for cell cycle. Apoptotic cells were quantified by flow cytometry analysis for cell apoptosis. The interplay between miR-423-5p and circ_FURIN or MTM1 was identified by dual-luciferase reporter and RNA pull-down assays. Results Circ_FURIN and MTM1 expressions were significantly upregulated, whereas miR-423-5p was downregulated in the ovarian cortex tissues of PCOS patients and TTR-treated KGN cells compared with controls. Circ_FURIN depletion relieved TTR-induced proliferation inhibition and apoptosis promotion. Besides, knockdown of miR-423-5p, a target miRNA of circ_FURIN, rescued circ_FURIN knockdown-mediated effects under TTR treatment. MiR-423-5p remitted TTR-induced cell disorders by binding to MTM1. Moreover, circ_FURIN modulated MTM1 expression through miR-423-5p. Conclusion Circ_FURIN silencing protected against TTR-induced dysfunction by the miR-423-5p/MTM1 pathway in human ovarian granulosa-like tumor cells.

The development trend of power converter is high frequency, high efficiency and high-power density. Compared with silicon-based power devices, SiC MOSFET has been widely used because of its fast-switching speed, low switching loss and high operating temperature. However, voltage spikes and oscillations are easy to occur during the high-speed turn-off of devices, which will seriously threaten the safe operation of power electronic converters. To solve this problem, based on the detailed analysis of the device turn-off process, a closed-loop active gate driver circuit based on dv/dt voltage feedback is proposed. Based on the small signal analysis of control oriented MOSFET driver circuit, the stability of dv/dt control is studied, and the influence of key parameters of driver circuit on the stability of control system is analyzed. The experimental results show that compared with the conventional gate driver, the proposed active gate driver can effectively suppress the peak voltage in the turn-off process without significantly increasing the switching loss.

Plants and algae associate with microbial communities that affect their growth and development. Here, the authors characterize the microbiota associated with a unicellular alga in soil, revealing extensive taxonomic and functional overlap with the root microbiota of land plants.

BackgroundHuman papillomavirus (HPV) infection is an important carcinogenic infection highly prevalent among many populations. However, independent influencing factors and predictive models for HPV infection in both U.S. and Beijing females are rarely confirmed. In this study, our first objective was to explore the overlapping HPV infection-related factors in U.S. and Beijing females. Secondly, we aimed to develop an R package for identifying the top-performing prediction models and build the predictive models for HPV infection using this R package.MethodsThis cross-sectional study used data from the 2009–2016 NHANES (a national population-based study) and the 2019 data on Beijing female union workers from various industries. Prevalence, potential influencing factors, and predictive models for HPV infection in both cohorts were explored.ResultsThere were 2,259 (NHANES cohort, age: 20–59 years) and 1,593 (Beijing female cohort, age: 20–70 years) participants included in analyses. The HPV infection rate of U.S. NHANES and Beijing females were, respectively 45.73 and 8.22%. The number of male sex partners, marital status, and history of HPV infection were the predominant factors that influenced HPV infection in both NHANES and Beijing female cohorts. However, condom application was not an independent influencing factor for HPV infection in both cohorts. R package Modelbest was established. The nomogram developed based on Modelbest package showed better performance than the nomogram which only included significant factors in multivariate regression analysis.ConclusionCollectively, despite the widespread availability of HPV vaccines, HPV infection is still prevalent. Compared with condom promotion, avoidance of multiple sexual partners seems to be more effective for preventing HPV infection. Nomograms developed based on Modelbest can provide improved personalized risk assessment for HPV infection. Our R package Modelbest has potential to be a powerful tool for future predictive model studies.

Abstract Background Heterotopic ossification (HO) represents pathological lesions that refer to the development of heterotopic bone in extraskeletal tissues around joints. This study investigates the genetic characteristics of bone marrow mesenchymal stem cells (BMSCs) from HO tissues and explores the potential pathways involved in this ailment. Methods Gene expression profiles (GSE94683) were obtained from the Gene Expression Omnibus (GEO), including 9 normal specimens and 7 HO specimens, and differentially expressed genes (DEGs) were identified. Then, protein–protein interaction (PPI) networks and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed for further analysis. Results In total, 275 DEGs were differentially expressed, of which 153 were upregulated and 122 were downregulated. In the biological process (BP) category, the majority of DEGs, including EFNB3, UNC5C, TMEFF2, PTH2, KIT, FGF13, and WISP3, were intensively enriched in aspects of cell signal transmission, including axon guidance, negative regulation of cell migration, peptidyl-tyrosine phosphorylation, and cell-cell signaling. Moreover, KEGG analysis indicated that the majority of DEGs, including EFNB3, UNC5C, FGF13, MAPK10, DDIT3, KIT, COL4A4, and DKK2, were primarily involved in the mitogen-activated protein kinase (MAPK) signaling pathway, Ras signaling pathway, phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) signaling pathway, and Wnt signaling pathway. Ten hub genes were identified, including CX3CL1, CXCL1, ADAMTS3, ADAMTS16, ADAMTSL2, ADAMTSL3, ADAMTSL5, PENK, GPR18, and CALB2. Conclusions This study presented novel insight into the pathogenesis of HO. Ten hub genes and most of the DEGs intensively involved in enrichment analyses may be new candidate targets for the prevention and treatment of HO in the future.

Hypopharyngeal squamous cell carcinoma (HSCC) is a kind of head and neck squamous cell carcinoma (HNSCC) with poor prognosis. Metabolic reprogramming may regulate the tumor microenvironment (TME) by adapting quickly to cellular stress and regulating immune response, but its role in HSCC has not been reported. We used the nCounter® Metabolic Pathways Panel to investigate metabolic reprogramming, cellular stress, and their relationship in HSCC tissues and adjacent normal tissues. Metabolism-related pathways nucleotide synthesis and glycolysis pathways were significantly upregulated, while amino acid synthesis and fatty acid oxidation pathways were significantly downregulated in HSCC tissues compared to adjacent normal tissues. There is a significant correlation between metabolism-related pathways and cellular stress pathways. Enrichment of immune cell and tumor infiltrating lymphocyte (TIL) analysis showed changes in immune responses between HSCC tissues and adjacent normal tissues. Overall survival analysis showed that upregulated genes CD276, LDHB, SLC3A2, EGFR, SLC7A5, and HPRT1 are potential unfavorable prognostic markers in HNSCC, while downregulated genes EEA1, IDO1, NCOA2, REST, CCL19, and ALDH2 are potential favorable prognostic markers in HNSCC. Moreover, metabolism-related genes IDO1, ALDH2, NCOA2, SLC7A5, SLC3A2, LDHB, and HPRT1 are correlated with immune infiltrates in HNSCC. These results suggest that metabolic reprogramming occurs and correlates with cellular stress and immune response in HSCC, which may help researchers understand mechanisms of metabolic reprogramming and develop effective immunotherapeutic strategies in HNSCC.

In machining 508III steel, the cemented carbide tool is subjected to a strong periodic thermal load impact, leading to serious tool-chip adhesion and shortening the tool life. Considering the influence of cutting parameters on temperature, temperature experiments and finite element (FE) simulations were carried out based on Box-Behnken experimental design criteria in the response surface method (RSM). Based on the experimental results, A second-order polynomial regression prediction model for temperature was constructed as the optimization objective function based on RSM. A temperature prediction model based on GA-SVM was established to predict temperature change. Taking cutting temperature and efficiency as evaluation indicators, the elitist nondominated sorting genetic algorithm was used to optimize cutting parameters. These findings may be important for the tool life improvement and reasonable parameter selection.

Research has shown that the lipid microenvironment surrounding colorectal cancer (CRC) is closely associated with the occurrence, development, and metastasis of CRC. According to pathological images from the National Center for Tumor diseases (NCT), the University Medical Center Mannheim (UMM) database and the ImageNet data set, a model called VGG19 was pre-trained. A deep convolutional neural network (CNN), VGG19CRC, was trained by the migration learning method. According to the VGG19CRC model, adipose tissue scores were calculated for TCGA-CRC hematoxylin and eosin (H&E) images and images from patients at Zhujiang Hospital of Southern Medical University and First People's Hospital of Chenzhou. Kaplan-Meier (KM) analysis was used to compare the overall survival (OS) of patients. The XCell and MCP-Counter algorithms were used to evaluate the immune cell scores of the patients. Gene set enrichment analysis (GSEA) and single-sample GSEA (ssGSEA) were used to analyze upregulated and downregulated pathways. In TCGA-CRC, patients with high-adipocytes (high-ADI) CRC had significantly shorter OS times than those with low-ADI CRC. In a validation queue from Zhujiang Hospital of Southern Medical University (Local-CRC1), patients with high-ADI had worse OS than CRC patients with low-ADI. In another validation queue from First People's Hospital of Chenzhou (Local-CRC2), patients with low-ADI CRC had significantly longer OS than patients with high-ADI CRC. We developed a deep convolution network to segment various tissues from pathological H&E images of CRC and automatically quantify ADI. This allowed us to further analyze and predict the survival of CRC patients according to information from their segmented pathological tissue images, such as tissue components and the tumor microenvironment.

ABSTRACT Plant roots are colonized by microorganisms from the surrounding soil that belong to different kingdoms and form a multikingdom microbial community called the root microbiota. Despite their importance for plant growth, the relationship between soil management, the root microbiota, and plant performance remains unknown. Here, we characterize the maize root-associated bacterial, fungal, and oomycetal communities during the vegetative and reproductive growth stages of four maize inbred lines and the pht1;6 phosphate transporter mutant. These plants were grown in two long-term experimental fields under four contrasting soil managements, including phosphate-deficient and -sufficient conditions. We showed that the maize root-associated microbiota is influenced by soil management and changes during host growth stages. We identified stable bacterial and fungal root-associated taxa that persist throughout the host life cycle. These taxa were accompanied by dynamic members that covary with changes in root metabolites. We observed an inverse stable-to-dynamic ratio between root-associated bacterial and fungal communities. We also found a host footprint on the soil biota, characterized by a convergence between soil, rhizosphere, and root bacterial communities during reproductive maize growth. Our study reveals the spatiotemporal dynamics of the maize root-associated microbiota and suggests that the fungal assemblage is less responsive to changes in root metabolites than the bacterial community. IMPORTANCE Plant roots are inhabited by microbial communities called the root microbiota, which supports plant growth and health. We show in a maize field study that the root microbiota consists of stable and dynamic members. The dynamics of the microbial community appear to be driven by changes in the metabolic state of the roots over the life cycle of maize.

Precision cancer medicine necessitates a personalized treatment plan for each individual patient. Given cancer’s heterogeneity and dynamic nature, the plot of patient-specific signatures composed of multiple cancer circulating biomarkers is useful to reveal the complete tumor landscape for guiding precision medicine. As an emerging new technology, surface-enhanced Raman scattering (SERS) shows the intrinsic advantage of performing multiplexed detection with the extremely narrow Raman spectral line widths. In this review, we first discuss the design principle of SERS nanotags to enable the detection of multiple circulating biomarkers, highlighting the important roles of plasmonic nanostructures and triple bond-modulated Raman reporters. Following this, we detail the use of isotropic and anisotropic nanostructures as SERS enhancement substrates for amplifying Raman signals in multi-biomarker detection. Furthermore, we present the triple bond-modulated molecules as Raman reporters in SERS nanotags to expand the multiplexing capability for biomarker measurements. Finally, we offer critical insights into the challenges and perspectives of SERS nanotags for cancer diagnosis, particularly from the aspect of future clinical transition. It is expected that this review can facilitate the design of more functional SERS nanotags with high sensitivity and multiplexing capability to assist early and accurate cancer screening. We also believe our review will be of interest in the fields of molecular imaging, biomedicine, and analytical chemistry.

In order to find out the changes of flavor substances in the processing of Colla corii asini (Ejiao) and provide reference for the flavor and quality control in the production of Ejiao, gas chromatography-ion mobility spectrometry (GC-IMS) was used to analyze the Ejiao products in different processing stages and establish the fingerprint. The differences among these stages were analyzed using multivariate statistical analysis, and the mechanism underlying volatile flavor compound formation was explored by discriminant analysis of Ejiao at different processing stages. The results indicated that Ejiao contains 47 volatile flavor compounds at different processing stages; they mainly include aldehydes, alcohols, esters, ketones, dimethyl disulfide, thiazole, and pyrazines. During Ejiao processing, the formation of these substances is mainly attributable to the Maillard reaction, amino acid or protein-oxidized lipid interaction, lipid oxidation and degradation, and long-chain compound degradation during heating. Principal component analysis results showed that volatile flavor compounds could be used to distinguish different Ejiao processing stages. The current results provide some reference for flavor and quality control of Ejiao products.

Small-signal stability analysis of the voltage source converter (VSC) based multi-terminal DC (MTDC) grids has been mainly focused on the dynamics of AC/DC converters while there is a lack of consideration of DC networks with DC power-flow controllers. In this paper, an integrated small-signal model is established to investigate the interactions between VSC and DC current flow controllers (CFC). The impacts of the control system of CFC on small-signal stability of the integrated system and the impacts of the control system of VSC on the DC networks with CFC are discussed via eigenvalue analysis. The above interactions are demonstrated through dynamic simulations against both small and large disturbances using real time digital simulator (RTDS).

With the completion of the relocation task, fully promoting the non-agricultural employment of relocated women is of great significance for improving the livelihood sustainability of relocated peasant households’ and consolidating the results of poverty alleviation. Based on the sample data of 1616 rural households in southern Shaanxi Province, China, using the IV-Probit model, IV-Tobit model, and mediation effect model, this study empirically analyzes the impact and mechanism of participation in relocation on women’s non-agricultural employment. The results show that by introducing an instrumental variable to solve endogeneity, participation in relocation has a significant positive impact on the behavior and intensity of women’s non-agricultural employment. A mechanism analysis shows that training, public services, and land abandonment play a mediation role in the relationship between participation in relocation and women’s non-agricultural employment. In terms of women’s non-agricultural employment behavior, the mediation effects are 56.9%, 15.0%, and 11.0%, respectively. In terms of women’s non-agricultural employment intensity, the mediation effects are 58.5%, 15.7%, and 11.6%, respectively. Based on the above findings, this study puts forward policy implications for follow-up relocation support, to further release the surplus women’s labor forces and promote women’s non-agricultural employment.

Difficulty in protoplast regeneration is a major obstacle to apply the CRISPR/Cas9 gene editing technique effectively in research and breeding of rapeseed (Brassica napus L.). The present study describes for the first time a rapid and efficient protocol for the isolation, regeneration and transfection of protoplasts of rapeseed cv. Kumily, and its application in gene editing. Protoplasts isolated from leaves of 3–4 weeks old were cultured in MI and MII liquid media for cell wall formation and cell division, followed by subculture on shoot induction medium and shoot regeneration medium for shoot production. Different basal media, types and combinations of plant growth regulators, and protoplast culture duration on each type of media were investigated in relation to protoplast regeneration. The results showed that relatively high concentrations of NAA (0.5 mg l−1) and 2,4-D (0.5 mg l−1) in the MI medium were essential for protoplasts to form cell walls and maintain cell divisions, and thereafter auxin should be reduced for callus formation and shoot induction. For shoot regeneration, relatively high concentrations of cytokinin were required, and among all the combinations tested, 2.2 mg l−1 TDZ in combination with auxin 0.5 mg l−1 NAA gave the best result with up to 45% shoot regeneration. Our results also showed the duration of protoplast culture on different media was critical, as longer culture durations would significantly reduce the shoot regeneration frequency. In addition, we have optimized the transfection protocol for rapeseed. Using this optimized protocol, we have successfully edited the BnGTR genes controlling glucosinolate transport in rapeseed with a high mutation frequency.

This paper aims to solve the problem of high reseeding rates and mis-seeding rates in the rice multi-grain hole direct seeding process. A multi grain cluster air suction type rice hill direct seed metering device was developed, and its seeding mechanism was analyzed. Based on CFD-DEM coupling simulation and bench tests, this study explored and optimized the performance of the seed metering device, and carried out the seeding adaptability test. The simulation results were as follows: when the negative pressure was −5 kPa, the static pressure, dynamic pressure and velocity of the flow field reached the maximum. When the negative pressure was −4 kPa, the qualification index was 89.62%, the reseeding index was 4.36%, and the mis-seeding index was 6.02%. The results of the orthogonal rotation combination test of three factors and five levels showed that when the rotation speed, negative pressure and the length of stirring brush were 20.70 rpm, −4.0 kPa and 10.50 mm respectively, the seed metering performance was the best, the qualification index was 90.85%, the reseeding index was 4.41% and the mis-seeding index was 4.74%. The seed metering device had strong adaptability to the sowing of different rice varieties, and met the agronomic requirements of direct seeding and seeding in rice holes.

Age is a potential predictive marker for the prognosis of cancer patients treated with immune checkpoint inhibitors (ICIs), but the appropriate age cutoff point is still controversial. We aimed to explore the influence of different age cutoff points on the prediction of prognosis for patients receiving ICIs and explore the mechanism underlying the appropriate age cutoff point from the aspects of gene mutation and expression, immune cell infiltration and so on. We applied cutoff points of 50, 55, 60, 65, 70, and 75 years old to divide 1660 patients from the Memorial Sloan-Kettering Cancer Center (MSKCC) immunotherapy cohort into older and younger groups and performed survival analysis of the six subgroups. The results showed that older patients had better survival than younger patients in accordance with the cutoff point of 50 years old [median overall survival (OS) (95% CI): 13.0 (10.5-15.5) months vs. 20.0 (16.7-23.3) months; p=0.002; unadjusted hazard ratio (HR) (95% CI): 0.77 (0.65-0.91)], whereas no significant difference was observed with other cutoff points. Further analysis of The Cancer Genome Atlas (TCGA) database and the MSKCC immunotherapy cohort data showed that the tumor mutation burden (TMB), neoantigen load (NAL), DNA damage response and repair (DDR) pathway mutation status, mutation frequencies of most genes (except IDH1, BRAF and ATRX), the expression of most immune-related genes and the degree of infiltration of most immune cells (such as CD8+ T cells and M1 macrophages) were higher in the elderly group (aged ≥50 years).

Abstract Aims This real‐world study is conducted to evaluate the efficacy and safety of recombinant human endostatin (rh‐endostatin) combined with chemotherapy as first‐line treatment for non‐driver genes mutation non‐small cell lung cancer (NSCLC) patients, and establish evidence‐based optimal regimen for rh‐endostatin. Patients and Methods Using propensity score matching (cut‐off: 0.01), 88 patients were eligible for our study, 34 of which received platinum‐based chemotherapy alone (chemotherapy group), 54 patients received platinum‐based chemotherapy plus rh‐endostatin (rh‐endostatin group). Among those 54 patients in the rh‐endostatin group, 27 patients received rh‐endostatin administered at 7.5 mg/m2 from day 1 to day 14 (rh‐endostatin 14d group), and the other 27 patients were administered at 15 mg/m2 from day 1 to day 7 (rh‐endostatin 7d group). The primary endpoint was progression‐free survival (PFS) and secondary endpoints were overall survival (OS), overall response rate (ORR), disease control rate (DCR), and safety. Results There were no differences in clinic characteristics among 3 groups. Compared with chemotherapy group, rh‐endostatin group improved PFS and OS significantly. The median PFS was 6 months vs 4.5 months (P = 0.047), and median OS was 20 months vs 10 months (P

Ischemia/reperfusion of organ systems in trauma patients with resuscitated hemorrhagic shock (HSR) contributes to tissue injury and organ dysfunction. Previous studies using a murine model of HSR showed that remote ischemic preconditioning (RIC) protected against organ injury and that the plasma was able to prevent neutrophil migration in a zebrafish tailfin-cut inflammation model. In this study, we hypothesized that RIC plasma inhibits neutrophil function through a decrease in reactive oxygen species (ROS) production via the upregulation of the transcription factor Nrf2 and downstream antioxidative genes. Plasma from mice subjected to RIC (4 cycles of 5-min hindlimb ischemia/reperfusion) was microinjected into zebrafish. The results show that RIC plasma caused a reduction of ROS generation in response to tail injury. In addition, RIC plasma protected the fish larvae in the survival studies when exposed to either H2O2 or LPS. Oxidative stress PCR Array showed that RIC plasma treatment led to upregulation of antioxidative related genes including hsp70, hmox1a, nqo1 as well as downregulation of duox, the producer of H2O2. To explore the role of nrf2 in RIC, RIC plasma from Nrf2 KO mice were injected to the zebrafish and showed no inhibitory effect on neutrophil migration. Moreover, knockdown of nrf2a attenuated the anti-inflammatory and protective effect of RIC plasma. The downregulation of duox and upregulation of hmox1a were confirmed to require the activation of nrf2a. Therefore, we show that the protective effect of RIC may be related to the elaboration of humoral factors which counter injury-induced ROS generation in a nrf2-dependent fashion.

Preparations from Arabian Gulf catfish (Arius bilineatus, Val) epidermal gel secretion (PCEGS) effectively heal chronic wounds in diabetic patients. However, specific lipid components of PCEGS that are responsible for various aspects of wound healing are unknown. Here, we report for the first time that, i) a unique preparation containing only proteins and lipids (Fraction B, FB), derived from the PCEGS accelerated the healing of experimental dermal wounds in female rats (transdermal punch biopsy) in vivo. Histological analyses showed that topical treatment of these wounds with FB promoted the migration of fibroblasts, facilitated the production of extracellular matrix (collagen, fibronectin), induced capillary formation and recruitment of immune cells, and accelerated overall wound healing by day 4 (tested at 1, 2, 3, 4, and 10 days; n=15 for vehicle; n=15 for FB treatment), ii) the lipids responsible for different stages of wound healing were separated into a protein-free bioactive lipid fraction, Ft, which contained a few common long-chain fatty acids, a unique furan fatty acid (F6) and a cholesterol metabolite, cholesta-3,5-diene (S5). Ft (the partially purified lipid fraction of PCEGS), and F6 and S5 present in Ft, proved to be bioactive for wound healing in human dermal fibroblasts. Ft increased the production and extracellular deposition of collagen and fibronectin, ex vivo, iii) Ft and its subcomponents, pure F6 and S5, also promoted human dermal fibroblast migration into the scratch wound gaps, ex vivo, iv) Ft, F6, and S5 promoted the recruitment of neutrophils (Green fluorescence protein labeled) to the site of injury in the transected tailfins of transgenic zebrafish, in vivo, v) Ft, but not F6 or S5, promoted the regeneration of tissues at the wound site in the transgenic zebrafish tailfin, in vivo. Therefore, we conclude that lipid fraction Ft from PCEGS contains the components necessary to promote complete wound healing, and F6 and S5 are responsible for promoting fibroblast and neutrophil recruitment to the site of wounds.

Abstract Assessing the CRC subtypes that can predict the outcome of colorectal cancer (CRC) in patients with immunogenicity seems to be a promising strategy to develop new drugs that target the antitumoral immune response. In particular, the disinhibition of the antitumoral T‐cell response by immune checkpoint blockade has shown remarkable therapeutic promise for patients with mismatch repair (MMR) deficient CRC. In this review, the authors provide the update of the molecular features and immunogenicity of CRC, discuss the role of possible predictive biomarkers, illustrate the modern immunotherapeutic approaches, and introduce the most relevant ongoing preclinical study and clinical trials such as the use of the combination therapy with immunotherapy. Furthermore, this work is further to understand the complex interactions between the immune surveillance and develop resistance in tumor cells. As expected, if the promise of these developments is fulfilled, it could develop the effective therapeutic strategies and novel combinations to overcome immune resistance and enhance effector responses, which guide clinicians toward a more “personalized” treatment for advanced CRC patients.

Mei (Prunus mume) is a woody tree that produces ornamental blossoms which symbolize spring in East Asia. Here, Zhang et al. resequence wild and domesticated mei to reveal considerable admixture and introgression from other Prunus species and identify loci associated with floral traits.

Their optical clarity as larvae and embryos, small size, and high fecundity make zebrafish ideal for whole animal high throughput screening. A high-throughput drug discovery platform (HTP) has been built to perform fully automated screens of compound libraries with zebrafish embryos. A Tg(kdrl:EGFP) line, marking endothelial cell cytoplasm, was used in this work to help develop protocols and functional algorithms for the system, with the intent of screening for angiogenesis inhibitors. Indirubin 3' Monoxime (I3M), a known angiogenesis inhibitor, was used at various concentrations to validate the protocols. Consistent with previous studies, a dose dependant inhibitory effect of I3M on angiogenesis was confirmed. The methods and protocols developed here could significantly increase the throughput of drug screens, while limiting human errors. These methods are expected to facilitate the discovery of novel anti-angiogenesis compounds and can be adapted for many other applications in which samples have a good fluorescent signal.

Identifying novel anti-cancer drugs is important for devising better cancer treatment options. In a series of studies designed to identify novel therapeutic compounds, we recently showed that a C-20 fatty acid (12,15-epoxy-13,14-dimethyleicosa-12,14-dienoic acid, a furanoic acid or F-6) present in the lipid fraction of the secretions of the Arabian Gulf catfish skin (Arius bilineatus Val.; AGCS) robustly induces neutrophil extracellular trap formation. Here, we demonstrate that a lipid mix (Ft-3) extracted from AGCS and F-6, a component of Ft-3, dose dependently kill two cancer cell lines (leukemic K-562 and breast MDA MB-231). Pure F-6 is approximately 3.5 to 16 times more effective than Ft-3 in killing these cancer cells, respectively. Multiplex assays and network analyses show that F-6 promotes the activation of MAPKs such as Erk, JNK, and p38, and specifically suppresses JNK-mediated c-Jun activation necessary for AP-1-mediated cell survival pathways. In both cell lines, F-6 suppresses PI3K-Akt-mTOR pathway specific proteins, indicating that cell proliferation and Akt-mediated protection of mitochondrial stability are compromised by this treatment. Western blot analyses of cleaved caspase 3 (cCasp3) and poly ADP ribose polymerase (PARP) confirmed that F-6 dose-dependently induced apoptosis in both of these cell lines. In 14-day cell recovery experiments, cells treated with increasing doses of F-6 and Ft-3 fail to recover after subsequent drug washout. In summary, this study demonstrates that C-20 furanoic acid F-6, suppresses cancer cell proliferation and promotes apoptotic cell death in leukemic and breast cancer cells, and prevents cell recovery. Therefore, F-6 is a potential anti-cancer drug candidate.

Correct specification of the left-right (L-R) axis is important for organ morphogenesis. Conserved mechanisms involving cilia rotation inside node-like structures and asymmetric Nodal signaling in the lateral plate mesoderm (LPM), which are important symmetry-breaking events, have been intensively studied. In zebrafish, the clustering and migration of dorsal forerunner cells (DFCs) is critical for the formation of the Kuppfer's vesicle (KV). However, molecular events underlying DFC clustering and migration are less understood. The WD-repeat proteins function in a variety of biological processes, including cytoskeleton assembly, intracellular trafficking, mRNA splicing, transcriptional regulation and cell migration. However, little is known about the function of WD-repeat proteins in L-R asymmetry determination. Here, we report the identification and functional analyses of zebrafish wdr18, a novel gene that encodes a WD-repeat protein that is highly conserved among vertebrate species. wdr18 was identified from a Tol2 transposon-mediated enhancer trap screen. Follow-up analysis of wdr18 mRNA expression showed that it was detected in DFCs or the KV progenitor cells and later in the KV at early somitogenesis stages. Morpholino knockdown of wdr18 resulted in laterality defects in the visceral organs, which were preceded by the mis-expression of Nodal-related genes, including spaw and pitx2. Examination of morphants at earlier stages revealed that the KV had fewer and shorter cilia which are immotile and a smaller cavity. We further investigated the organization of DFCs in wdr18 morphant embryos using ntl and sox17 as specific markers and found that the clustering and migration of DFC was altered, leading to a disorganized KV. Finally, through a combination of wdr18 and itgb1b morpholino injections, we provided evidence that wdr18 and itgb1b genetically interact in the laterality determination process. Thus, we reveal a new and essential role for WD-repeat proteins in the determination and regulation of L-R asymmetry and propose a potential mechanism for wdr18 in the regulation of DFC clustering and migration and KV formation.

Macrosegregation is a critical factor that limits the mechanical properties of materials. The impact of equiaxed crystal sedimentation on macrosegregation has been extensively studied, as it plays a significant role in determining the distribution of alloying elements and impurities within a material. To improve macrosegregation in steel connecting shafts, a multiphase solidification model that couples melt flow, heat transfer, microstructure evolution, and solute transport was established based on the volume-averaged Eulerian-Eulerian approach. In this model, the effects of liquid phase, equiaxed crystals, columnar dendrites, and columnar-to-equiaxed transition (CET) during solidification and evolution of microstructure can be considered simultaneously. The sedimentation of equiaxed crystals contributes to negative macrosegregation, where regions between columnar dendrites and equiaxed crystals undergo significant A-type positive macrosegregation due to the CET. Additionally, noticeable positive macrosegregation occurs in the area of final solidification in the ingot. The improvement in macrosegregation is beneficial for enhancing the mechanical properties of connecting shafts. To mitigate the thermal convection of molten steel resulting from excessive superheating, reducing the superheating during casting without employing external fields or altering the design of the ingot mold is indeed an effective approach to control macrosegregation. [ABSTRACT FROM AUTHOR]