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Abstract Low back pain (LBP) is largely attributed to intervertebral disc degeneration (IVDD), of which the endplate changes are an important component. However, the alterations in cell fate and properties within the endplates during degeneration remain unknown. Here, we firstly performed the single-cell RNA-sequencing analysis (scRNA-seq) of the cells focusing on degenerative human endplates. By unsupervised clustering of the 8,534 single-cell based on the gene expression, we identified nine distinct cell types. We employed Gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis, and the single-cell regulatory network inference and clustering (SCENIC) to determine the enriched pathways and transcriptional activities across seven chondrocyte subpopulations. Furthermore, two cell fates of chondrocyte differentiation were found by trajectory analysis, one was enriched in inflammation-related genes, and the other was related to extracellular matrix (ECM). Additionally, the intercellular interactions of macrophages (MA) and chondrocytes, T cells/natural killer cells (T/NK) and chondrocytes were examined by ligand-receptor pairs analysis, showing the important regulative function of FN1 from MA and CD74 from T/NK during endplate degeneration. Overall, our findings provide novel perspectives on the endplate degeneration at the single-cell level and a whole-transcriptome size.

Abstract Citrullination plays an essential role in various physiological or pathological processes, however, whether citrullination is involved in regulating tumour progression and the potential therapeutic significance have not been well explored. Here, we find that peptidyl arginine deiminase 4 (PADI4) directly interacts with and citrullinates hypoxia-inducible factor 1α (HIF-1α) at R698, promoting HIF-1α stabilization. Mechanistically, PADI4-mediated HIF-1αR698 citrullination blocks von Hippel-Lindau (VHL) binding, thereby antagonizing HIF-1α ubiquitination and subsequent proteasome degradation. We also show that citrullinated HIF-1αR698, HIF-1α and PADI4 are highly expressed in hepatocellular carcinoma (HCC) tumour tissues, suggesting a potential correlation between PADI4-mediated HIF-1αR698 citrullination and cancer development. Furthermore, we identify that dihydroergotamine mesylate (DHE) acts as an antagonist of PADI4, which ultimately suppresses tumour progression. Collectively, our results reveal citrullination as a posttranslational modification related to HIF-1α stability, and suggest that targeting PADI4-mediated HIF-1α citrullination is a promising therapeutic strategy for cancers with aberrant HIF-1α expression.

ABSTRACT Background Exosomes have been established to be enriched with various long noncoding RNAs (lncRNAs) and circular RNAs (circRNAs) that exert various biological effects. However, the lncRNA‐ and circRNA‐mediated coexpression competing endogenous RNA (ceRNA) regulatory network in exosomes derived from the plasma of patients with lung adenocarcinoma (LUAD) remains elusive. Methods and Results This study enrolled nine patients with lung adenocarcinoma and three healthy individuals, and the differential expression of messenger RNAs (mRNAs), lncRNAs, and circRNAs was detected using microarray analysis, while microRNAs (miRNAs) were detected through RNA sequencing. Additionally, bioinformatics algorithms were applied to evaluate the lncRNA–miRNA–mRNAs/circRNA–miRNA–mRNA network. Differentially expressed cicRNAs were identified via quantitative reverse transcription polymerase chain reaction (RT‐qPCR). A total of 1016 lncRNAs, 1396 circRNAs, 45 miRNAs, and 699 mRNAs were differentially expressed in the plasma exosomes of patients with LUAD compared with healthy controls. Among them, 881 lncRNAs were upregulated and 135 were downregulated, 916 circRNAs were upregulated while 480 were downregulated, 45 miRNAs were upregulated while none were downregulated, and 591 mRNAs were upregulated while 108 were downregulated (p ≤ 0.05, and fold change ≥ 2). Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed the biological functions of differentially expressed RNAs. Meanwhile, the RNA networks displayed the regulatory relationship between dysregulated RNAs. Finally, RT‐qPCR validated that the expression of circ‐0033861, circ‐0043273, and circ‐0011959 was upregulated in the plasma exosome of patients with LUAD compared to healthy controls (p = 0.0327, p = 0.0002, p = 0.0437, respectively). Conclusion This study proposed a newly discovered ncRNA–miRNA–mRNA/circRNA–miRNA–mRNA ceRNA network and identified that the expression of circulating circ‐0033861, circ‐0043273, and circ‐0011959 was up‐regulated in the plasma exosomes of patients with LUAD, offering valuable insights for exploring the potential function of exosomal noncoding RNA and identifying potential biomarkers for LUAD.

Plant basic helix-loop-helix (bHLH) transcription factors (TFs) play central roles in various abiotic stresses. However, its role in plant cold resistance is largely unknown. Previously, we characterised CaNAC035 in pepper, which positively regulates tolerance to cold, salt and drought stresses tolerance. Here, we identified CabHLH035, a CaNAC035-interacting protein in pepper. To explore its functions in cold stress tolerance, we silenced the gene in pepper via virus-induced gene silencing (VIGS) and overexpressed the gene in Arabidopsis. The results showed that CabHLH035 expression was induced by cold treatment, and silencing of CabHLH035 decreased cold stress tolerance. Conversely, overexpression of CabHLH035 in Arabidopsis increased cold stress tolerance. To investigate homologs genes of C-repeat binding factor (CBF) pathway proteins and reactive oxygen species (ROS) marker gene expression blocking by CabHLH035, we performed yeast one-hybrid (Y1H), dual luciferase and electrophoretic mobility shift assay experiments. The results showed that CabHLH035 bound to the region upstream of the CaCBF1A and CaAPX promoters. Additionally, CaCBF1A bound to the CaDHN4 promoter. Taken together, our results showed that CabHLH035 plays a crucial role in cold stress tolerance and its potential as a target for breeding cold-resistant crops. The findings provide a basis for studying the functions and regulatory network of cold stress tolerance in pepper.

Abstract Objective Clinically, it has been found that patients undergoing knee replacement have a high incidence of concomitant hallux valgus. In this study, we analyzed whether patients with osteoarthritis who underwent surgery and those patient who did not have surgery had an increased risk of hallux valgus by Mendelian randomization and performed reverse causal analysis. Design Genomewide association study (GWAS) data for osteoarthritis, categorized by knee arthritis with joint replacement, knee arthritis without joint replacement, hip arthritis with joint replacement, and hip arthritis without joint replacement.And acquired hallux valgus were downloaded for Mendelian randomized studies. MR analysis was performed using inverse variance-weighted (IVW), weighted median, and MR-Egger methods. MR-egger regression, MR pleiotropic residuals and outliers (MR-presso), and Cochran's Q statistical methods were used to evaluate heterogeneity and pleiotropy. Results The IVW results indicate that, compared to healthy individuals, patients who meet the criteria for knee osteoarthritis joint replacement surgery have a significantly higher risk of acquired hallux valgus. There were no significant causal relationships found for the remaining results. No significant heterogeneity or multiplicity was observed in all the Mr analyses. Conclusion Our study supports the increased risk of acquired hallux valgus in patients eligible for knee replacement. There is necessary for clinicians to be concerned about the hallux valgus status of patients undergoing knee arthroplasty.

Abstract Endplate sclerosis is a notable aspect of spine degeneration or aging, but the mechanisms remain unclear. Here, we report that senescent macrophages accumulate in the sclerotic endplates of lumbar spine instability (LSI) or aging male mouse model. Specifically, knockout of cdkn2a (p16) in macrophages abrogates LSI or aging-induced angiogenesis and sclerosis in the endplates. Furthermore, both in vivo and in vitro studies indicate that IL-10 is the primary elevated cytokine of senescence-related secretory phenotype (SASP). Mechanistically, IL-10 increases pSTAT3 in endothelial cells, leading to pSTAT3 directly binding to the promoters of Vegfa, Mmp2, and Pdgfb to encourage their production, resulting in angiogenesis. This study provides information on understanding the link between immune senescence and endplate sclerosis, which might be useful for therapeutic approaches.

Objective Glucocorticoid (GC) overuse is strongly associated with steroid‐induced osteonecrosis of the femoral head (SINFH). However, the underlying mechanism of SINFH remains unclear. This study aims to investigate the effect of dexamethasone (Dex)‐induced oxidative stress on osteocyte apoptosis and the underlying mechanisms. Methods Ten patients with SINFH and 10 patients with developmental dysplasia of the hips (DDH) were enrolled in our study. Sixty rats were randomly assigned to the Control, Dex, Dex + N‐Acetyl‐L‐cysteine (NAC), Dex + Dibenziodolium chloride (DPI), NAC, and DPI groups. Magnetic resonance imaging (MRI) was used to examine edema in the femoral head of rats. Histopathological staining was performed to assess osteonecrosis. Immunofluorescence staining with TUNEL and 8‐OHdG was conducted to evaluate osteocyte apoptosis and oxidative damage. Immunohistochemical staining was carried out to detect the expression of NOX1, NOX2, and NOX4. Viability and apoptosis of MLO‐Y4 cells were measured using the CCK‐8 assay and TUNEL staining. 8‐OHdG staining was conducted to detect oxidative stress. 2′,7′‐Dichlorodihydrofluorescein diacetate (DCFH‐DA) staining was performed to measure reactive oxygen species (ROS). The expression of NOX1, NOX2, and NOX4 in MLO‐Y4 cells was analyzed by Western blotting. Multiple comparisons were performed using one‐way analysis of variance (ANOVA). Results In patients and the rat model, hematoxylin–eosin (HE) staining revealed a significantly higher rate of empty lacunae in the SINFH group than in the DDH group. Immunofluorescence staining indicated a significant increase in TUNEL‐positive cells and 8‐OHdG‐positive cells in the SINFH group compared to the DDH group. Immunohistochemical staining demonstrated a significant increase in the expression of NOX1, NOX2, and NOX4 proteins in SINFH patients compared to DDH patients. Moreover, immunohistochemical staining showed a significant increase in the proportion of NOX2‐positive cells compared to the Control group in the femoral head of rats. In vitro, Dex significantly inhibited the viability of osteocyte cells and induced apoptosis. After Dex treatment, the intracellular ROS level increased. However, Dex treatment did not alter the expression of NOX proteins in vitro. Additionally, NAC and DPI inhibited the generation of intracellular ROS and partially alleviated osteocyte apoptosis in vivo and in vitro. Conclusion This study demonstrates that GC promotes apoptosis of osteocyte cells through ROS‐induced oxidative stress. Furthermore, we found that the increased expression of NOXs induced by GC serves as an important source of ROS generation.

Summary: OXCT1 acts as a succinyltransferase to promote serine beta-lactamase-like protein (LACTB) K284 succinylation. Here, we present a protocol for detecting OXCT1-mediated LACTB succinylation levels and sites. We describe steps for using western blotting (WB) and mass spectrometry to determine OXCT1-mediated LACTB succinylation levels and sites in vitro. This protocol can be applied to detect and identify succinylation levels and sites on other proteins.For complete details on the use and execution of this protocol, please refer to Ma et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract Immune checkpoint blockade (ICB) has shown considerable promise for treating various malignancies, but only a subset of cancer patients benefit from immune checkpoint inhibitor therapy because of immune evasion and immune-related adverse events (irAEs). The mechanisms underlying how tumor cells regulate immune cell response remain largely unknown. Here we show that hexokinase domain component 1 (HKDC1) promotes tumor immune evasion in a CD8+ T cell-dependent manner by activating STAT1/PD-L1 in tumor cells. Mechanistically, HKDC1 binds to and presents cytosolic STAT1 to IFNGR1 on the plasma membrane following IFNγ-stimulation by associating with cytoskeleton protein ACTA2, resulting in STAT1 phosphorylation and nuclear translocation. HKDC1 inhibition in combination with anti-PD-1/PD-L1 enhances in vivo T cell antitumor response in liver cancer models in male mice. Clinical sample analysis indicates a correlation among HKDC1 expression, STAT1 phosphorylation, and survival in patients with hepatocellular carcinoma treated with atezolizumab (anti-PD-L1). These findings reveal a role for HKDC1 in regulating immune evasion by coupling cytoskeleton with STAT1 activation, providing a potential combination strategy to enhance antitumor immune responses.

Abstract Mechanical force contributes to perforin pore formation at immune synapses, thus facilitating the cytotoxic T lymphocytes (CTL)-mediated killing of tumor cells in a unidirectional fashion. How such mechanical cues affect CTL evasion of perforin-mediated autolysis remains unclear. Here we show that activated CTLs use their softness to evade perforin-mediated autolysis, which, however, is shared by T leukemic cells to evade CTL killing. Downregulation of filamin A is identified to induce softness via ZAP70-mediated YAP Y357 phosphorylation and activation. Despite the requirements of YAP in both cell types for softness induction, CTLs are more resistant to YAP inhibitors than malignant T cells, potentially due to the higher expression of the drug-resistant transporter, MDR1, in CTLs. As a result, moderate inhibition of YAP stiffens malignant T cells but spares CTLs, thus allowing CTLs to cytolyze malignant cells without autolysis. Our findings thus hint a mechanical force-based immunotherapeutic strategy against T cell leukemia.

Abstract Mitochondrial rRNA modifications are essential for mitoribosome assembly and its proper function. The m4C methyltransferase METTL15 maintains mitochondrial homeostasis by catalyzing m4C839 located in 12 S rRNA helix 44 (h44). This modification is essential to fine-tuning the ribosomal decoding center and increasing decoding fidelity according to studies of a conserved site in Escherichia coli. Here, we reported a series of crystal structures of human METTL15–hsRBFA–h44–SAM analog, METTL15–hsRBFA–SAM, METTL15–SAM and apo METTL15. The structures presented specific interactions of METTL15 with different substrates and revealed that hsRBFA recruits METTL15 to mitochondrial small subunit for further modification instead of 12 S rRNA. Finally, we found that METTL15 deficiency caused increased reactive oxygen species, decreased membrane potential and altered cellular metabolic state. Knocking down METTL15 caused an elevated lactate secretion and increased levels of histone H4K12-lactylation and H3K9-lactylation. METTL15 might be a suitable model to study the regulation between mitochondrial metabolism and histone lactylation.

Abstract Itaconate is a well-known immunomodulatory metabolite; however, its role in hepatocellular carcinoma (HCC) remains unclear. Here, we find that macrophage-derived itaconate promotes HCC by epigenetic induction of Eomesodermin (EOMES)-mediated CD8+ T-cell exhaustion. Our results show that the knockout of immune-responsive gene 1 (IRG1), responsible for itaconate production, suppresses HCC progression. Irg1 knockout leads to a decreased proportion of PD-1+ and TIM-3+ CD8+ T cells. Deletion or adoptive transfer of CD8+ T cells shows that IRG1-promoted tumorigenesis depends on CD8+ T-cell exhaustion. Mechanistically, itaconate upregulates PD-1 and TIM-3 expression levels by promoting succinate-dependent H3K4me3 of the Eomes promoter. Finally, ibuprofen is found to inhibit HCC progression by targeting IRG1/itaconate-dependent tumor immunoevasion, and high IRG1 expression in macrophages predicts poor prognosis in HCC patients. Taken together, our results uncover an epigenetic link between itaconate and HCC and suggest that targeting IRG1 or itaconate might be a promising strategy for HCC treatment.

Pantana phyllostachysae Chao is a leaf-eating pest that poses a significant threat to bamboo forest health. Current research mainly focuses on statically identifying damage using remote sensing images. However, the mechanism behind the damage's traceability remains unclear, making it difficult to pinpoint early infestation sources accurately. Additionally, our understanding of the pest's spreading laws is limited. This study leverages Sentinel-2A/B images from February to November 2021 to investigate P. phyllostachysae infestation traceability through the dynamic age algorithm and indicator analysis method. The results shed light on the distribution of early pest sources over the study period. By analyzing both the overall pest infestation “cluster” and its center of gravity, we dissect P. phyllostachysae infestation characteristics and paths monthly throughout the study period. Our findings reveal three zones with strong spreading momentum, three with slow spreading momentum, and two transitional zones during the February-November period, aligning with P. phyllostachysae occurrence patterns. However, the direction of P. phyllostachysae spreading varies, likely due to a combination of meteorological, topographical, vegetative biochemical, and human activity factors. This study introduces innovative approaches for identifying early pest source points and understand their spreading laws, contributing to more effective pest prevention and control in forest ecosystems.

Abstract Background The relationship between aging and osteoporosis is well established. However, the relationship between the body's physiological age, i.e. epigenetic age, and osteoporosis is not known. Our goal is to analyze the bidirectional causal relationship between epigenetic clocks and osteoporosis using a bidirectional Mendelian randomization study. Methods We used SNPs closely associated with GrimAge, Hannum, PhenoAge, and HorvathAge in epigenetic age and SNPs closely associated with femoral neck bone mineral density, lumbar spine bone mineral density, and forearm bone mineral density as instrumental variables, respectively, using the inverse variance weighting method and several other MR methods to assess the bidirectional causal relationship between epigenetic age and osteoporosis. Result There was no evidence of a clear causal relationship of epigenetic age (GrimAge, Hannum, PhenoAge, and HorvathAge) on femoral neck bone mineral density, lumbar spine bone mineral density, and forearm bone mineral density. In reverse Mendelian randomization analysis showed a significant causal effect of lumbar spine bone mineral density on GrimAge: odds ratio (OR) = 0.692, 95% confidence interval (CI) = (0.538–0.890), p = 0.004. The results suggest that a decrease in lumbar spine bone mineral density promotes an acceleration of GrimAge. Conclusion There was no significant bidirectional causal relationship between epigenetic age and osteoporosis A decrease in lumbar spine bone density may lead to an acceleration of the epigenetic clock "GrimAge". Our study provides partial evidence for a bidirectional causal effect between epigenetic age and Osteoporosis.

Abstract Background Postoperative delirium (POD) is an acute form of brain dysfunction that can result in serious adverse consequences. There has been a link between cognitive dysfunction and poor sleep. The present study aimed to determine the association and prediction of subjective sleep quality and postoperative delirium during major non-cardiac surgery. Methods One hundred and thirty-four patients, aged 60 years or older, were scheduled for elective laparotomy or orthopaedic procedures. The Pittsburgh Sleep Quality Index (PSQI) and sleep log were used to assess perioperative subjective sleep quality in participants. Nursing Delirium Screening Checklist (NU-DESC) was used for screening, and the Confusion Assessment Method (CAM) was used to diagnose POD during the first seven days following surgery. The association between subjective sleep quality and POD was assessed using a multivariate logistic regression model. Thereafter, the prediction performance of subjective sleep quality was evaluated using a receiver operating characteristic (ROC) curve. Results All assessments were completed on 119 patients who had an average PSQI score of 7.0 ± 2.4 before surgery. 23 patients (19.3%) suffered from POD. The multivariate logistic regression analysis showed that the occurrence of POD was closely related to age, BMI, PSQI and operation time. After adjusting for related factors, there was a statistically significant association between PSQI and POD occurrence (OR = 1.422, 95%CI 1.079–1.873, per 1-point increase in PSQI). The ROC curve analysis showed that the optimal PSQI cutoff value was 8.0 for predicting POD, and the area under the ROC (AUROC) value of PSQI was 0.741 (95%CI 0.635 to 0.817). The AUROC of the model developed by the multivariate logistic regression analysis was 0.870 (95%CI 0.797 to 0.925). Conclusions The study found that preoperative subjective sleep quality was strongly associated with POD during major non-cardiac surgery. Additionally, PSQI combined with age, BMI, and operation time improved POD prediction.

Moso bamboo (Phyllostachys pubescens) stands as a pivotal economic bamboo species globally, holding substantial potential for carbon sequestration. Accurate estimation of aboveground biomass (AGB) in Moso bamboo forests is crucial due to its close ties with the ecosystem's carbon cycle. Despite the maturation of monitoring techniques for Pantana phyllostachysae Chao, a significant pest of Moso bamboo, its interplay with AGB in these forests remains enigmatic. This study addressed this gap by categorizing P. phyllostachysae's impact on Moso bamboo forests into four levels: healthy, mild damage, moderate damage, and severe damage. By scrutinizing field data, we delved into the shifts in Moso bamboo leaf biomass under P. phyllostachysae stress. Leveraging Sentinel-2A/B imagery, we extracted diverse correlation factors, including original wave bands, vegetation indices, texture attributes, and vegetation's physical and chemical parameters. Subsequently, machine learning algorithms-namely, random forest (RF), support vector machine (SVM), and extreme gradient boosting (XGB) were employed to achieve remote sensing inversion of AGB in Moso bamboo forests, accounting for the presence of insect pests. We analyzed the response of Moso bamboo biomass sensitive factors and to further clarify the changes of AGB of Moso bamboo forests under insect pest stress at the remote sensing level ultimately. The results showed that (1) the degree of Moso bamboo leaf biomass damage was positively related to the damage level, which gradually increased from 15.15 % to 59.42 %; (2) the RF algorithm excelled in estimating Moso bamboo forest AGB, particularly in May, and inclusion of insect pest considerations enhanced AGB estimation accuracy; (3) among the four factor types, Band information and vegetation indices emerged as most impactful, and Band5, Band11, Band12, NDVI68a and MSAVI were selected the most often; (4) at the remote sensing level, AGB in Moso bamboo forests significantly varies under P. phyllostachysae stress. Healthy areas demonstrate an AGB of 66.9037 Mg ha−1, while heavily affected regions drop to 52.6591 Mg ha−1. It can be seen that combining pest factors for Moso bamboo biomass estimation solves the problem of rough biomass estimation, and this study provides a more promising method for forest growth monitoring.

Bamboo groves predominantly thrive in tropical or subtropical regions. Assessing the efficacy of remote sensing data of various types in extracting bamboo forest information from bright and shadow areas is a critical issue for achieving precise identification of bamboo forests in complex terrain. In this study, 34 features were obtained from Sentinel-1 SAR and Sentinel-2 optical images using the Google Earth Engine platform. The normalized shaded vegetation index (NSVI) was then employed to segment the bright and shadow woodlands. Different features from diverse data sources were evaluated to extract bamboo forest information in the bright and shadow areas, then use the random forest (RF) classification algorithm to extract bamboo forest. The results showed that (1) the red-edge and short-wave infrared bands of Sentinel-2 optical images and their corresponding vegetation indices are significant in bamboo forest information extraction. (2) The dissimilarity and homogeneity of Sentinel-2 texture features in the bright area and dissimilarity in the shadow area, the Sentinel-1 backscatter features in the bright area and the VV and VH in the bright area and VV-VH in the shadow area have some variability between bamboo and nonbamboo forests, which can be used as effective features for bamboo forest extraction. (3) The combination of spectral, texture and backscatter features yields the highest overall classification accuracy and Kappa coefficient, at 87.96% and 0.7435, respectively. This study has the potential for remote sensing refinement of bamboo forest identification in complex terrain areas by utilizing subregion classification methods combined with optical and radar image features.

The secretome of hepatocytes and hepatocellular carcinoma (HCC) cells can contribute to cancer progression. Here the authors show that PRSS35 inhibits HCC progression through proteolytic depletion of CXCL2 and subsequently decreased neutrophil recruitment to tumours.

Abstract The ribosomal protein contains complex structures that belong to polypeptide glycoprotein family, which are involved in plant growth and responses to various stresses. In this study, we found that capsicum annuum 40S ribosomal protein SA-like (CaSLP) was extensively accumulated in the cell nucleus and cell membrane, and the expression level of CaSLP was up-regulated by Salicylic acid (SA) and drought treatment. Significantly fewer peppers plants could withstand drought stress after CaSLP gene knockout. The transient expression of CaSLP leads to drought tolerance in pepper, and Arabidopsis’s ability to withstand drought stress was greatly improved by overexpressing the CaSLP gene. Exogenous application of SA during spraying season enhanced drought tolerance. CaSLP-knockdown pepper plants demonstrated a decreased resistance of Pseudomonas syringae PV.tomato (Pst) DC3000 (Pst.DC3000), whereas ectopic expression of CaSLP increased the Pst.DC3000 stress resistance in Arabidopsis. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) results showed that CaNAC035 physically interacts with CaSLP in the cell nucleus. CaNAC035 was identified as an upstream partner of the CaPR1 promoter and activated transcription. Collectively the findings demonstrated that CaSLP plays an essential role in the regulation of drought and Pst.DC3000 stress resistance. Graphical Abstract

The objective of this study was to deeply understand the adaptation mechanism of the functional traits of Moso bamboo Phyllostachys pubescens syn. edulis (Poales: Poaceae) leaves to the environment under different Pantana phyllostachysae Chao damage levels, analyzing the changes in the relationship between specific leaf area (SLA) and leaf dry matter content (LDMC). We combined different machine learning models (decision tree, RF, XGBoost, and CatBoost regression models), and used different canopy heights and different levels of infestation, to analyze the changes in the relationship between the two under different levels of infestation based on the results of the best estimation model. The results showed the following: (1) The SLA of Ph. pubescens showed a decreasing trend with the increase om insect pest degree, and LDMC showed an inverse trend. (2) The SLA of bamboo leaves was negatively correlated with the LDMC under different insect pest degrees; the correlation of the data under the healthy class was higher than that of other insect pest levels, and at the same time better than that of the full sample, which laterally confirmed the effect of insect pest stress on the functional traits of Ph. pubescens leaves. (3) When modeling under different infestation levels, the CatBoost model was used for heavy damage and the RF model was used for the rest of the cases; the decision tree regression model was used when modeling different canopy heights. The findings contribute certain insights into the nuanced responses and adaptive mechanisms of Ph. pubescens forests to environmental fluctuations. Moreover, these results furnish a robust scientific foundation, essential for ensuring the enduring sustainability of Ph. pubescens forest ecosystems.

In order to improve the real-time performance of gesture recognition by a micro-Doppler map of mmWave radar, the point cloud based gesture recognition for mmWave radar is proposed in this paper. Two steps are carried out for mmWave radar-based gesture recognition. The first step is to estimate the point cloud of the gestures by 3D-FFT and the peak grouping. The second step is to train the TRANS-CNN model by combining the multi-head self-attention and the 1D-convolutional network so as to extract the features in the point cloud data at a deeper level to categorize the gestures. In the experiments, TI mmWave radar sensor IWR1642 is used as a benchmark to evaluate the feasibility of the proposed approach. The results show that the accuracy of the gesture recognition reaches 98.5%. In order to prove the effectiveness of our approach, a simply 2Tx2Rx radar sensor is developed in our lab, and the accuracy of recognition reaches 97.1%. The results show that our proposed gesture recognition approach achieves the best performance in real time with limited training data in comparison with the existing methods.

The purpose of this meta‐analysis was to identify if patient‐specific instrumentation (PSI) could increase the accuracy of the correction in high tibial osteotomy (HTO) and to explore the assessment indices and the necessity of using a PSI in HTO. A systematic search was carried out using online databases. A total of 466 patients were included in 11 papers that matched the inclusion criteria. To evaluate the accuracy of PSI‐assisted HTO, the weight bearing line ratio (WBL%), hip‐knee‐ankle angle (HKA), mechanical medial proximal tibial angle (mMPTA), and posterior tibial slope angle (PTSA) were measured preoperatively and postoperatively and compared to the designed target values. Statistical analysis was performed after strict data extraction with Review Manager (version 5.4). Significant differences were detected in WBL% (MD = −36.41; 95% CI: −42.30 to −30.53; p

Objective: In order to provide evidence to scientific exploitation of Yaowang tea endemic to Qinling mountain, nutritional value, health benefits and their material basis of Yaowang tea were investigated. Methods: Chemical composition as well as nutrients and bioactive constituents profile of Yaowang tea was analyzed by high performance liquid chromatography and plasma emission spectroscopy, etc. Antioxidant activity was determined by free radical scavenging experiment, and protective effect on DNA damage was studied by agarose gel eletrophoresis. Results: Quercitrin, quercetin and gallic acid were determined in Yaowang tea, and their contents were 0.17%±0.00%, 0.29%±0.01% and 0.28%±0.01%, respectively. Content of essential amino acids was 65.65±1.97 mg/g. Contents of manganese, selenium and magnesium were 1.19±0.04, 0.43±0.02 and 2.80±0.05 mg/kg, respectively, while heavy metals (e.g., arsenic, cadmium, mercury, lead) did not exceed the national permissible limit. Yaowang tea extract exhibited the ability to scavenge DPPH and ABTS radicals with EC50 values of 0.0704 and 0.3810 mg/mL, respectively. Protective activity of Yaowang tea extract against DNA damage was in dose-dependent manner. Protective effect of 1×10−2 mg/mL Yaowang tea extract against DNA damage was higher than that of ascorbic acid. Conclusion: Yaowang tea contains several bioactive constituents, and it is safe in respect to heavy metals. Yaowang tea possesses in vitro antioxidant activity and protective effect against DNA damage.

Abstract Macrophages in tumors (tumor-associated macrophages, TAMs), a major population within most tumors, play key homeostatic functions by stimulating angiogenesis, enhancing tumor cell growth, and suppressing antitumor immunity. Resetting TAMs by simple, efficacious and safe approach(s) is highly desirable to enhance antitumor immunity and attenuate tumor cell malignancy. Previously, we used tumor cell-derived microparticles to package chemotherapeutic drugs (drug-MPs), which resulted in a significant treatment outcome in human malignant pleural effusions via neutrophil recruitments, implicating that drug-MPs might reset TAMs, considering the inhibitory effects of M2 macrophages on neutrophil recruitment and activation. Here, we show that drug-MPs can function as an antitumor immunomodulator by resetting TAMs with M1 phenotype and IFN-β release. Mechanistically, drug molecules in tumor MPs activate macrophage lysosomal P450 monooxygenases, resulting in superoxide anion formation, which further amplifies lysosomal ROS production and pH value by activating lysosomal NOX2. Consequently, lysosomal Ca2+ signaling is activated, thus polarizing macrophages towards M1. Meanwhile, the drug molecules are delivered from lysosomes into the nucleus where they activate DNA sensor hnRNPA2B1 for IFN-β production. This lysosomal-nuclear machinery fully arouses the antitumor activity of macrophages by targeting both lysosomal pH and the nuclear innate immunity. These findings highlight that drug-MPs can act as a new immunotherapeutic approach by revitalizing antitumor activity of macrophages. This mechanistic elucidation can be translated to treat malignant ascites by drug-MPs combined with PD-1 blockade.

Type 2 diabetes mellitus (T2DM) has become a worldwide public health problem. Epimedin C is considered one of the most important flavonoids in Epimedium, a famous edible herb in China and Southeast Asia that is traditionally used in herbal medicine to treat diabetes. In the present study, the therapeutic potential of epimedin C against T2DM was ascertained using a mouse model, and the mechanism underlying the hypoglycemic activity of epimedin C was explored using a label-free proteomic technique for the first time. Levels of fasting blood glucose (FBG), homeostasis model assessment of insulin resistance (HOMA-IR), and oral glucose tolerance, as well as contents of malondialdehyde (MDA) and low-density lipoprotein cholesterol (LDL-C) in the 30 mg·kg−1 epimedin C group (EC30 group), were significantly lower than those in the model control group (MC group) (p < 0.05), while the contents of hepatic glycogen, insulin, and high-density lipoprotein cholesterol (HDL-C), as well as activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the EC30 group were notably higher than those in the MC group (p < 0.05). The structures of liver cells and tissues were greatly destroyed in the MC group, whereas the structures of cells and tissues were basically complete in the EC30 group, which were similar to those in the normal control group (NC group). A total of 92 differentially expressed proteins (DEPs) were enriched in the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. In the EC30 vs. MC groups, the expression level of cytosolic phosphoenolpyruvate carboxykinase (Pck1) was down-regulated, while the expression levels of group XIIB secretory phospholipase A2-like protein (Pla2g12b), apolipoprotein B-100 (Apob), and cytochrome P450 4A14 (Cyp4a14) were up-regulated. According to the KEGG pathway assay, Pck1 participated in the gluconeogenesis and insulin signaling pathways, and Pla2g12b, Apob, and Cyp4a14 were the key proteins in the fat digestion and fatty acid degradation pathways. Pck1, Pla2g12b, Apob, and Cyp4a14 seemed to play important roles in the prevention and treatment of T2DM. In summary, epimedin C inhibited Pck1 expression to maintain FBG at a relatively stable level, promoted Pla2g12b, Apob, and Cyp4a14 expressions to alleviate liver lipotoxicity, and protected liver tissues and cells from oxidant stress possibly by its phenolic hydroxyl groups.

IntroductionThe causal relationship between Coronavirus disease 2019 (COVID-19) and osteoporosis (OP) remains uncertain. We aimed to assess the effect of COVID-19 severity (severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, COVID-19 hospitalization, and severe COVID-19) on OP by a two-sample Mendelian randomization (MR) study.MethodsWe conducted a two-sample MR analysis using publicly available genome-wide association study (GWAS) data. Inverse variance weighting (IVW) was used as the main analysis method. Four complementary methods were used for our MR analysis, which included the MR–Egger regression method, the weighted median method, the simple mode method, and the weighted mode method. We utilized the MR-Egger intercept test and MR pleiotropy residual sum and outlier (MR-PRESSO) global test to identify the presence of horizontal pleiotropy. Cochran’s Q statistics were employed to assess the existence of instrument heterogeneity. We conducted a sensitivity analysis using the leave-one-out method.ResultsThe primary results of IVW showed that COVID-19 severity was not statistically related to OP (SARS-CoV-2 infection: OR (95% CI) = 0.998 (0.995 ~ 1.001), p = 0.201403; COVID-19 hospitalization: OR (95% CI) =1.001 (0.999 ~ 1.003), p = 0.504735; severe COVID-19: OR (95% CI) = 1.000 (0.998 ~ 1.001), p = 0.965383). In addition, the MR-Egger regression, weighted median, simple mode and weighted mode methods showed consistent results. The results were robust under all sensitivity analyses.ConclusionThe results of the MR analysis provide preliminary evidence that a genetic causal link between the severity of COVID-19 and OP may be absent.

BackgroundThe Achilles tendon is the strongest and most susceptible tendon in humans. Achilles tendon injuries and ruptures have gradually attracted research attention. However, a bibliometric analysis of global research in this field is lacking. This study involved a bibliometric analysis of the developmental trends and research hotspots in Achilles tendon injuries/ruptures from 2000 to 2021.MethodsArticles published between 2001 and 2021 were retrieved from an extended database of the Science Citation Index using Web of Science. VOSviewer and CiteSpace were used to analyze the relationships between publications, countries, institutions, journals, authors, references, and keywords.ResultsThis study included 3,505 studies of 73 countries, 3,274 institutions, and 12,298 authors and explored the cooperation between them and the relationships between citations. Over the past 22 years, the number of publications has significantly increased. Foot Ankle International has published the most papers on Achilles tendon injuries/ruptures, and British Journal of Sports Medicine is the most famous journal. Re-rupture, exosomes, acute Achilles tendon rupture, and tendon adhesions gradually become the research focus over the past few years.ConclusionAchilles tendon injury and rupture are important research topics. A vast number of newly published papers on this topic have demonstrated that clinicians and researchers are interested in their study. Over time, these recent studies will be widely cited; therefore, this bibliometric analysis should be constantly updated.

ObjectiveThis study aimed to investigate the relationship of the K-line with sagittal cervical curvature changes and surgical outcomes in patients with cervical ossification of the posterior longitudinal ligament (OPLL).MethodsWe retrospectively reviewed 84 patients with OPLL who underwent posterior cervical single-door laminoplasty. The patients were divided into a K-line-positive (+) group and a K-line-negative (−) group. Perioperative data, radiographic parameters, and clinical outcomes were compared between the two groups.ResultsOf 84 total patients, 50 patients were in the K (+) group and 29 patients were in the K (–) group. Neurological function improved in both groups after laminoplasty. The C2–7 Cobb angle, T1 slope, and C2–7 sagittal vertical axis were significantly changed in the K(−) group compared with those in the K (+) group before the operation and at the 3-month and final follow-ups.ConclusionNeurological function was recovered in both groups, and the clinical effect on the K (+) group was better than that on the K (−) group. The cervical curvature in patients with OPLL tends to be anteverted and kyphotic after laminoplasty and is an important factor in reducing the clinical effect.

Significant progress has been achieved in universal basic education in African countries since the late 1990s. This study provides empirical evidence on the within- and across-country variation in numeracy skills performance among children based on nationally representative data from eight African countries (DR Congo, The Gambia, Ghana, Lesotho, Sierra Leone, Togo, Tunisia, and Zimbabwe). We assess whether and to what extent children with disabilities lag in numeracy skills and how much it depends on their type of disabilities. More specifically, we explore whether disabled children benefit equally from better school system quality. The assessment is analysed as a natural experiment using the performance of non-disabled children as a benchmark and considering the different types of disabilities as random treatments. We first evaluate the variation in average numeracy skills in the eight African countries. They can roughly be divided into low- and high-numeracy countries. We apply Instrumental Variable (IV) methods to control the endogeneity of completed school years when assessing subjects' school performance and heterogeneous disability effects. Children with vision and hearing disabilities are not especially challenged in numeracy skills performance. The low numeracy skills among physically and intellectually disabled children are mainly attributable to their limited school attendance. Children with multiple disabilities are constrained both by low school attendance and by poor numeracy skills return to schooling. The average differences in school performance across the high- versus low-numeracy skill country groups are larger than the within-group average differences for disabled versus non-disabled kids. This indicates that school enrolment and quality are crucial for children's learning of numeracy skills, and that disabled children benefit equally from better school quality across these African countries.

BackgroundOsteoarthritis has the characteristics of degenerative changes in articular cartilage and secondary osteoporosis, and it is a common chronic irreversible joint disease. In addition to affecting articular cartilage, subchondral bone, joint capsule and synovial membrane also undergo pathological changes during the development of the disease. Multiple studies have revealed that patients with osteoarthritis were found to have a significantly increased risk of osteoporosis, which also contributes to the progression of osteoarthritis. However, in the current existing studies, we found that no scholars have used bibliometric analysis in the study of the relationship between osteoarthritis and osteoporosis. From the perspective of bibliometrics, this study summarizes in detail the degree of cooperation between countries, research institutions, authors, and related journals in the field of osteoarthritis and osteoporosis research and their respective influence. In this way, the evolution of knowledge structure, the change of research focus and the hot topics with research potential in the future can be further visualized and analyzed.MethodsSearch the Web of Science core collection in Science Citation Index Expanded for articles and reviews of research on osteoarthritis and osteoporosis from 1998 to 2021. Bibliometric tools such as VOSviewer, CiteSpace, were be frequently used in our study. They are mainly used to analyze collaborations between countries, research institutions, and publication authors. Meantime, co-citation analysis of journals, co-occurrence analysis of keywords and subject categories will also be reflected in the study.ResultsAccording to the search strategy, 1,078 publications were included during the period 1998–2021. And the number of annual publications on the relationship between osteoarthritis and osteoporosis is on the rise. The United States has achieved the most and contributed the most in this field and the Boston University was the most prolific institution. For the statistical analysis of published publications, Reginster JY had the highest number of publications, while Felson DT had the highest co-citation frequency. Respectively, Osteoarthritis And Cartilage was the most productive journal in this area of research. The keywords “inflammation,” “expression,” and “mesenchymal stem cells” may also be the development trend and research hotspot of the future research direction in this field.ConclusionsIn our study, the relationship between osteoarthritis and osteoporosis was analyzed by using literature measurement. These analysis results can lead researchers to learn more directly about the trend in this area and provide guidance for determining popular research directions.

Abstract Metformin, the first-line drug for type II diabetes, has recently been considered an anticancer agent. However, the molecular target and underlying mechanism of metformin’s anti-cancer effects remain largely unclear. Herein, we report that metformin treatment increases the sensitivity of hepatocarcinoma cells to methotrexate (MTX) by suppressing the expression of the one-carbon metabolism enzyme DHFR. We show that the combination of metformin and MTX blocks nucleotide metabolism and thus effectively inhibits cell cycle progression and tumorigenesis. Mechanistically, metformin not only transcriptionally represses DHFR via E2F4 but also promotes lysosomal degradation of the DHFR protein. Notably, metformin dramatically increases the response of patient-derived hepatocarcinoma organoids to MTX without obvious toxicity to organoids derived from normal liver tissue. Taken together, our findings identify an important role for DHFR in the suppressive effects of metformin on therapeutic resistance, thus revealing a therapeutically targetable potential vulnerability in hepatocarcinoma.

Abstract Background Long noncoding RNAs (lncRNAs) are important regulators in tumor progression. However, their biological functions and underlying mechanisms in hypoxia adaptation remain largely unclear. Results Here, we established a correlation between a Chr3q29-derived lncRNA gene and tongue squamous carcinoma (TSCC) by genome-wide analyses. Using RACE, we determined that two novel variants of this lncRNA gene are generated in TSCC, namely LINC00887_TSCC_short (887S) and LINC00887_TSCC_long (887L). RNA-sequencing in 887S or 887L loss-of-function cells identified their common downstream target as Carbonic Anhydrase IX (CA9), a gene known to be upregulated by hypoxia during tumor progression. Mechanistically, our results showed that the hypoxia-augmented 887S and constitutively expressed 887L functioned in opposite directions on tumor progression through the common target CA9. Upon normoxia, 887S and 887L interacted. Upon hypoxia, the two variants were separated. Each RNA recognized and bound to their responsive DNA cis-acting elements on CA9 promoter: 887L activated CA9’s transcription through recruiting HIF1α, while 887S suppressed CA9 through DNMT1-mediated DNA methylation. Conclusions We provided hypoxia-permitted functions of two antagonistic lncRNA variants to fine control the hypoxia adaptation through CA9.

Abstract Background Histone deacetylases (HDACs) engage in the regulation of various cellular processes by controlling global gene expression. The dysregulation of HDACs leads to carcinogenesis, making HDACs ideal targets for cancer therapy. However, the use of HDAC inhibitors (HDACi) as single agents has been shown to have limited success in treating solid tumors in clinical studies. This study aimed to identify a novel downstream effector of HDACs to provide a potential target for combination therapy. Methods Transcriptome sequencing and bioinformatics analysis were performed to screen for genes responsive to HDACi in breast cancer cells. The effects of HDACi on cell viability were detected using the MTT assay. The mRNA and protein levels of genes were determined by quantitative reverse transcription‐PCR (qRT‐PCR) and Western blotting. Cell cycle distribution and apoptosis were analyzed by flow cytometry. The binding of CREB1 (cAMP‐response element binding protein 1) to the promoter of the KDELR (The KDEL (Lys‐Asp‐Glu‐Leu) receptor) gene was validated by the ChIP (chromatin immunoprecipitation assay). The association between KDELR2 and protein of centriole 5 (POC5) was detected by immunoprecipitation. A breast cancer‐bearing mouse model was employed to analyze the effect of the HDAC3‐KDELR2 axis on tumor growth. Results KDELR2 was identified as a novel target of HDAC3, and its aberrant expression indicated the poor prognosis of breast cancer patients. We found a strong correlation between the protein expression patterns of HADC3 and KDELR2 in tumor tissues from breast cancer patients. The results of the ChIP assay and qRT‐PCR analysis validated that HDAC3 transactivated KDELR2 via CREB1. The HDAC3‐KDELR2 axis accelerated the cell cycle progression of cancer cells by protecting the centrosomal protein POC5 from proteasomal degradation. Moreover, the HDAC3‐KDELR2 axis promoted breast cancer cell proliferation and tumorigenesis in vitro and in vivo. Conclusion Our results uncovered a previously unappreciated function of KDELR2 in tumorigenesis, linking a critical Golgi‐the endoplasmic reticulum traffic transport protein to HDAC‐controlled cell cycle progression on the path of cancer development and thus revealing a potential therapeutical target for breast cancer.

The carbon emission trading scheme (ETS) is an important measure to implement China’s “double carbon” strategy.We use “China’s carbon emission trading pilot policy” as a quasi-natural experiment to identify theeffect of this market-based environmental regulation on a firm’s export and its impacting mechanisms.Based on the Propensity score matching and difference-in-differences (PSM-DID) method, we observe robust evidence that the carbon emissions trading pilot policy significantly increases the export of regulated firms. And also find that this policy positivelyaffects the exports of both SOEs and non-SOEs. Considering enterprise heterogeneity, the policy positivelyimpacts the exports of FDI firms, large firms, and low industrial concentrations. Moreover, we examine how environmental regulation could affect firmexport through technological innovation, productivity, and product research. The observable evidence leads us to cautiously conclude thatmarket-based environmental regulations in even developing countries could achieve export growth.Based on our findings, we suggest that: 1) policymakers should limit CO2 emissions quotas to ensure an appropriate increase in the price of CO2 emissions; 2) to design a unified carbon ETS market, researchers should explore ways to activate market-oriented environmental regulation tools based on the carbon emission price.

ObjectiveTo investigate the association between the structural deformity and foot pain in hallux valgus (HV) patients using a multi-variate pattern analysis (MVPA) approach.MethodsPlain radiographic metrics were calculated from 36 painful and 36 pain-free HV feet. In analysis 1, univariate analyses were performed to investigate the clinical and radiographic differences between painful and pain-free HV. In analysis 2, we investigated the pattern differences for radiographic metrics between these two groups using a MVPA approach utilizing a support vector machine. In analysis 3, sequential backward selection and exhaustive search were performed as a feature-selection procedure to identify an optimal feature subtype. In analysis 4, hierarchical clustering analysis was used to identify the optimal radiographic HV subtype associated with pain in HV.ResultsWe found that: (1) relative to feet with pain-free HV, the painful ones exhibited a higher hallux valgus angle, i.e., the magnitude of distal metatarsal and phalangeal deviation; (2) painful HV could be accurately differentiated from pain-free HV via MVPA. Using sequential backward selection and exhaustive search, a 5-feature subset was identified with optimal performance for classifying HV as either painful or pain-free; and (3) by applying hierarchical clustering analysis, a radiographic subtype with an 80% pain incidence was identified.ConclusionThe pain in HV is multifactorial and associated with a radiographic pattern measured by various angles on plain radiographs. The combination of hallux valgus angle, inter-phalangeal angle, distal metatarsal articular angle, metatarsal cuneiform angle and metatarsal protrusion distance showed the optimal classification performance between painful and pain-free HV.

The m6A methylation is reported to function in multiple physiological and pathological processes. However, the functional relevance of m6A modification to post-spinal cord injured (SCI) damage is not yet clear. In the present study, methylated RNA immunoprecipitation combined with microarray analysis showed that the global RNA m6A levels were decreased following SCI. Then, gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) analyses were conducted to demonstrate the potential function of differential m6A-tagged transcripts and the altered transcripts with differential m6A levels. In addition, we found that the m6A “writer,” METTL3, significantly decreased after SCI in mice. The immunostaining validated that the expression of METTL3 mainly changed in GFAP or Iba-1+ cells. Together, this study shows the alteration of m6A modification following SCI in mice, which might contribute to the pathophysiology of the spinal cord after trauma.

Leaf water content (LWC) is very important in the growth of vegetation. LWC and leaf spectra change when the leaves are under pest stress; exploring the change mechanism between LWC, leaf spectra, and pest stress can lay the foundation for pest detection. In this study, we measured the LWC and leaf spectra of moso bamboo leaves under different damage levels, used the Pearson–Lasso method to screen the features, and established a multiple linear regression (MLR) and random forest regression (RFR) model to estimate the LWC. We analyzed the relationship between LWC and spectral features of moso bamboo leaves under Pantana phyllostachysae Chao (PPC) stress and their changes. The results showed that: (1) the LWC showed a decreasing trend as the pest level increased. (2) The spectra changed substantially when the leaves were under pest stress. (3) The number and significance of response features associated with the LWC were diverse under different damage levels. (4) The estimation of LWC under different damage levels differed significantly. LWC, leaf spectra, response features, and the model estimation effect were diverse under different damage levels. The correlation between LWC and features was higher for healthy leaves than for damaged and off-year leaves. The two models were more effective in estimating the LWC of healthy leaves but less effective for damaged and off-year leaves. This study provides theoretical support for the prediction of PPC stress and lays the foundation for remote sensing monitoring.

Abstract A novel planar distributed three-beam electron gun with narrow beam separations is designed based on grids loaded sheet beam method. The dimensions of the three-beam gun in the y–O-z plane are determined using our basic theoretical design method developed for sheet beam gun. The results show that the profile of focusing electrode in the y–O-z plane is related to the beam width in the x-O-z plane. Then, the characteristics and parameters of three-beam array formation with their stability are analyzed thoroughly by adjustment of control grids in the x-O-z plane. Each of the beamlet obtained is with a small axial deviation of the two transverse waists. Based on the theoretical analysis and simulations, the planar three-beam electron gun is constructed with the beam voltage of 22 kV and the current of 3 $$\times$$ × 0.15 A. The average radius of 0.08 mm at each beam waist is obtained with the compression factor of 4 for the 0.18 mm beam tunnel radius. The beam waist can be achieved at about 4.4 mm away from the cathode with the axis separation about 0.46 mm for each of beamlet. Thus, the design method can be generally used to construct such type of narrow beam separation and planar distributed multiple beam electron gun for the miniaturization and integrated vacuum electron devices in millimeter wave and terahertz band.

Abstract The health‐promoting benefits of medicine and food homology (MFH) are known for thousands of years in China. However, active compounds and biological mechanisms are unclear, greatly limiting clinical practice of MFH. The advent of gut microbiota analysis and metabolomics emerge as key tools to discover functional compounds, therapeutic targets, and mechanisms of benefits of MFH. Such studies hold great promise to promote and optimize functional efficacy and development of MFH‐based products, for example, foods for daily dietary supplements or for special medical purposes. In this review, we summarized pharmacological effects of 109 species of MFH approved by the Health and Fitness Commission in 2015. Recent studies applying genome sequencing of gut microbiota and metabolomics to explain the activity of MFH in prevention and management of health consequences were extensively reviewed. We discussed the potentiality in future to decipher functional activities of MFH by applying metabolomics‐based polypharmacokinetic strategy and multiomics technologies. The needs for personalized MFH recommendations and comprehensive databases have also been highlighted. This review emphasizes current achievements and challenges of the analysis of gut microbiota and metabolomics as a new avenue to understand MFH.

Glycogen can be metabolized via glycogenolysis and the pentose phosphate pathway as well as into the production of UDP glucose, which when secreted can bind the P2Y14 receptor. Here the authors show how these glycogen metabolism pathways contribute to proinflammatory macrophage activation and susceptibility to sepsis.

Aurora-A kinase is frequently over-expressed in tumours. Here, the authors show that it modulates the activity of lactate dehydrogenase B, resulting in enhanced glycolysis, bio-synthesis and tumour growth.

Beach-bar sand in lacustrine facies represents one of the most significant reservoirs. Depending on the depositional characteristics, it can be further divided into two different sedimentary microfacies, beach sand and the bar sand. Favorable reservoirs are often developed in bar sand. The lower section of the upper part of the 4th member of the Shahejie Formation in the Gao89-1 block is a typical nearshore shallow water beach-bar deposit. Oil distribution is influenced by lithofacies and physical properties. In order to better characterize the heterogeneity within beach-bar sandbodies, a modeling method based on the depositional mode and sandbody volume is proposed. Firstly, a sandbody model is established. On this basis, an algorithm for distinguishing between beach and bar sand based on vertical thickness is proposed. The model is post processed based on the sandbody volume to remove unreasonable sandbodies. The method allows for a more realistic three-dimensional geological model of the beach-bar sands in the study area than the classical two-point geostatistical, object-based, and multi-point simulation method. A facies-controlled modeling approach is used to establish a petrophysical property model on this foundation; the result shows that the property models better reflect the characteristics of the petrophysical distribution in the Gao89-1 block.

Prucalopride was widely used for chronic constipation, which is difficult to be adequately relieved by laxatives in adult patients in clinic. Due to the difficulty in metabolite identification, metabolic process of prucalopride had not been investigated in vivo. In this study, an efficient strategy was proposed for comprehensive metabolite profiling of prucalopride after oral administration in rat plasma, urine, and feces samples. This strategy was composed of five steps. First, the samples at multiple time points after oral administration were collected to increase the representativeness of the samples. Second, different sample preparation methods were investigated to obtain superior extraction efficiency. Third, the raw data of test sample and blank sample were acquired using ultra-performance liquid chromatography with Q-Exactive hybrid quadrupole–orbitrap high-resolution accurate mass spectrometry under the positive and negative full-scan/dd MS2 mode. Fourth, combined mass defect filter with background subtraction model in soft of compound discovery, all peaks were constructed to filter potential metabolites after retention time alignment and ion filtration, which could remove large amounts of interference ions. Besides, it can predict potential biotransformation, promoting to understand how to metabolize the drug. This provides multiple possibilities and prevents us conjecturing the potential metabolites blindly. Finally, the verification procedure was implemented through exporting the structure and MS2 spectrum to the analytical tool of Mass Frontier. The proposed strategy significantly improved the targeted detection and identification for metabolites in vivo. A total of 47 metabolites were tentatively characterized in the plasma, urine, and feces samples after oral administration of prucalopride. This study could provide a valuable reference for systematic metabolite profile of drug in vivo.

Acute spinal cord injury (ASCI) is a severe traumatic disease of the central nervous system, the underlying mechanism of which is unclear. This study was intended to study the role of EZH2 and miR-146a-5p/HIF-1α in inflammation and glycolysis after ASCI, providing reference and basis for the clinical treatment and prognosis of ASCI injury. We used lipopolysaccharide (LPS) to induce inflammation of microglia, and we constructed the ASCI animal model. qRT-PCR detected the relative expression levels of EZH2, HIF-1α, miR-146a-5p, IL-6, TNF-α, IL-17, PKM2, GLUT1, and HK2 in cells and tissues. Western blot was performed to detect the expression levels of EZH2, HIF-1α, H3K27me3, IL-6, TNF-α, IL-17, PKM2, GLUT1, and HK2. ChIP verified the enrichment of H3K27me3 in the miR-146a-5p promoter region. Bioinformatics predicted the binding sites of HIF-1α and miR-146a-5p, and dual-luciferase reporter assay verified the binding of HIF-1α and miR-146a-5p. ELISA detects the levels of inflammatory factors IL-6, TNF-α, and IL-17 in the cerebrospinal fluid of rats. The GC-TOFMS was used to detect the changes of glycolytic metabolites in the cerebrospinal fluid of rats. EZH2 could mediate inflammation and glycolysis of microglia. EZH2 regulates inflammation and glycolysis through HIF-1α. EZH2 indirectly regulated the HIF-1α expression by mediating miR-146a-5p. EZH2 mediates miR-146a-5p/HIF-1α to alleviate inflammation and glycolysis in ASCI rats. In the present study, our results demonstrated that EZH2 could mediate miR-146a-5p/HIF-1α to alleviate the inflammation and glycolysis after ASCI. Therefore, EZH2/miR-146a-5p/HIF-1α might be a novel potential target for treating ASCI.

NAC (NAM, ATAF1/2, and CUC2) proteins are the plant-specific transcription factors (TFs) which are important in plant response to abiotic stresses. However, knowledge about the functional role that NACs play in pepper abiotic stress tolerance is limited. In this study, we isolated a NAC TF gene, CaNAC035, from pepper (Capsicum annuum L.), where the protein is localized in the nucleus and functions as a transcriptional activator. CaNAC035 expression is induced by low and high temperatures, osmotic stress, salt, gibberellic acid (GA), methyl-jasmonic acid (MeJA), salicylic acid (SA), and abscisic acid (ABA). To understand the function of CaNAC035 in the abiotic stress responsep, we used virus-induced gene silencing in pepper to knockdown the CaNAC035 and overexpressed the CaNAC035 in Arabidopsis. The results showed that pepper seedlings in which CaNAC035 was silenced, showed more damage than the control pepper plants after cold, NaCl, and mannitol treatments. Correspondingly increased electrolyte leakage, a higher level of malondialdehyde (MDA), H2O2, and superoxide radicals were found after cold treatments. CaNAC035-silenced seedlings exhibited lower chlorophyll content while CaNAC035-overexpressed Arabidopsis plants had higher germination rate and fresh weight after mannitol and NaCl treatments. We also reported 18 proteins that potentially interact with CaNAC035 and may participate in processes such as the stress response, resistance, and photosynthesis. Our results suggest that CaNAC035 is a positive regulator of abiotic stress tolerance in pepper which acts through multiple signaling pathways.

Alcohol (ethanol) is one of the most common addictive psychoactive substances in the world, and alcoholism may result in harmful effects on human health, especially on the nervous system. Flavonoids are regarded as the main active constituent in Epimedium, which has been used to cure some nervous system diseases such as amnesia for over 1000 years. Here, the protective effects of Epimedium flavonoids against ethanol-induced toxicity in retinoic acid (RA)-treated SH-SY5Y cells were investigated. Their mechanism was explored by a label-free proteomic approach combined with bioinformatic analysis for the first time. The results showed that ethanol treatment decreased cell viability by 18%, whereas the viability increased significantly after intervention with Epimedium flavonoids (p < 0.01). According to proteomic and bioinformatic analyses, hundreds of differentially expressed proteins (DEPs) were identified and classified as biological process (GO_BP), cellular component (GO_CC) and molecular function (GO_MF). Among them, GO_MF of DEPs, especially molecular function relevant to G proteins, greatly changed in SH-SY5Y cells pretreated by Epimedium flavonoids. In the alcoholism pathway, the expression of the Gi protein was up-regulated under the influence of ethanol, whereas Epimedium flavonoids could reverse the expression profile, both of which were validated by Western blot assay. In conclusion, Gi protein seemed to be an important factor in the alcoholism pathway to suppress the ethanol-induced toxicity of SH-SY5Y cells. These findings suggest a protective potential of Epimedium flavonoids against ethanol-induced toxicity to neurons via the regulation of Gi protein function.

Cold stress is one of the main restricting factors affecting plant growth and agricultural production. Complex cold signaling pathways induce the expression of hundreds of cold-sensitive genes. The NAC transcription factor CaNAC035 has previously been reported to significantly influence the response of pepper to cold stress. Here, using Yeast one-hybrid (Y1H) library screened to search for other relevant molecular factors, we identified that CabHLH79 directly binds to the CaNAC035 promoter. Different basic helix–loop–helix (bHLH) transcription factors (TFs) in plants significantly respond to multiple plant stresses, but the mechanism of bHLHs in the cold tolerance of pepper is still unclear. This study investigated the functional characterization of CabHLH79 in the regulation of cold resistance in pepper. Down-regulation of CabHLH79 in pepper by virus-induced gene silencing (VIGS) increased its sensitivity to low temperature, whereas overexpression of CabHLH79 in pepper or Arabidopsis enhanced cold resistance. Compared with control plants, VIGS mediated of CabHLH79 had lower enzyme activity and related gene expression levels, accompanied by higher reactive oxygen species (ROS) accumulation, relative electrolyte leakage (REL), and malondialdehyde accumulation (MDA) contents. Transient overexpression of CabHLH79 pepper positively regulated cold stress response genes and ROS genes, which reduced REL and MDA contents. Similarly, ectopic expression of CabHLH79 in Arabidopsis showed less ROS accumulation, and higher enzymes activities and expression levels. These results indicated that CabHLH79 enhanced cold tolerance by enhancing the expression of ROS-related and other cold stress tolerance-related genes. Taken together, our results showed a multifaceted module of bHLH79-NAC035 in the cold stress of pepper.

Tumour-associated macrophages (TAMs) display an M2 phenotype that promote tumour immune escape. Here the authors show that Chloroquine (CQ), a lysosome inhibitor used against malaria, inhibits tumour growth by switching TAMs into an M1 tumor-killing phenotype by repolarizing macrophages metabolism.

Polo-like kinase 1 (Plk1) is a key regulator of mitosis. Here, the authors show that Plk1 activates the pentose phosphate pathway in cancer cells by directly phosphorylating glucose-6-phosphate dehydrogenase (G6PD) and that such activation is critical for cell cycle progression and cancer cell growth.

Menin is a protein with context-dependent oncogenic or oncosuppressive roles; the oncogenic activity is mainly due to its function as a cofactor of the MLL1 histone methyltransferase complex. Here the authors show that Menin regulates c-Myc-dependent transformation independently of the MLL complex.