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Abstract Pleiotropy is frequently detected in agronomic traits of wheat (Triticum aestivum). A locus on chromosome 4B, QTn/Ptn/Sl/Sns/Al/Tgw/Gl/Gw.caas-4B, proved to show pleiotropic effects on tiller, spike, and grain traits using a recombinant inbred line (RIL) population of Qingxinmai × 041133. The allele from Qingxinmai increased tiller numbers, and the allele from line 041133 produced better performances of spike traits and grain traits. Another 52 QTL for the eight traits investigated were detected on 18 chromosomes, except for chromosomes 5D, 6D, and 7B. Several genes in the genomic interval of the locus on chromosome 4B were differentially expressed in crown and inflorescence samples between Qingxinmai and line 041133. The development of the KASP marker specific for the locus on chromosome 4B is useful for molecular marker-assisted selection in wheat breeding.

Introduction Heart failure with preserved ejection fraction (HFpEF) is a prevalent comorbidity among patients with end-stage kidney disease. Although sodium-glucose cotransporter 2 inhibitors are validated in treating heart failure and ameliorating left ventricular hypertrophy among non-dialysis patients, the effects on dialysis patients are unknown. We previously investigated the pharmacokinetics of henagliflozin in patients undergoing haemodialysis (HD) or peritoneal dialysis (PD) and clarified its safety.Methods and analysis This multicentre, randomised, double-blind, placebo-controlled trial is being conducted at three hospitals in Shanghai, China. A target of 108 HD or PD patients with HFpEF are randomly allocated to treatment group (henagliflozin 5 mg/day in addition to standard therapy) or control group (placebo with standard therapy) at a ratio of 1:1. All subjects will be followed up for 24 weeks. The primary outcome is change in echocardiography-measured left ventricular mass index. The secondary interests include changes in left atrial volume index, E/e’, e’ and N-terminal pro-B-type natriuretic peptide (NT-proBNP). Intergroup comparisons of change in echocardiography-related outcomes from baseline to 24 weeks are based on a linear regression model adjusted for baseline values (analysis of covariance), and repeated measure analysis of variance with Bonferroni adjustment is employed for comparison of change in NT-proBNP. Subgroup analyses of the primary and secondary outcomes are conducted to determine whether the effect of henagliflozin varies according to dialysis modality. The χ2 method is used to compare the occurrence of adverse events and severe adverse events.Ethics and dissemination This trial has been approved by the Ethics Committee of Renji Hospital, School of Medicine, Shanghai Jiao Tong University (LY2023-127-B). All participants provide written informed consent before screening. The results of the trial will be disclosed completely in international peer-reviewed journals. Both positive and negative results will be reported.Trial registration number ChiCTR2300073169.

Most directed energy deposition (DED) implementations employ a rigid end effector to deposit material layer-by-layer, which is insufficient in cases with space constraints. To enhance the flexibility, this paper proposes a novel soft-robotic gun (SRG) based DED technology. The ideology is to replace the rigid weld gun with a tendon-driven omnidirectional-steering continuum manipulator, which a computer can numerically control. To this end, both requirements and concepts of the SRG-DED are introduced. A prototype system is designed and implemented before dedicated kinematic modelling and trajectory planning mechanisms are presented. As demonstrated by the experimental results, the developed trajectory planning algorithm improves the computational efficiency of tendon variables and enhances the consistency of trajectory tracking. The prototype system can deposit a complex path on an inclined substrate and fabricate a thin-wall component within a pipeline. The validation work confirms the feasibility and applicability of the prototype system for metal additive manufacturing applications.

ABSTRACTIn multi-robot cooperative wire and arc additive manufacturing (MRC-WAAM) systems, allocating and scheduling deposition tasks is necessary. The challenge comes from the factorial computational complexity when optimising robot time, workload, and deposition order. This work presents a novel method, ‘top k%’. The main idea is to iteratively assign k% amount of paths from a warehouse to the robot having the least workload until all paths are set. A collision-free deposition process is scheduled by checking the overlapping conditions of robot swap areas. Both simulative testing and actual depositions were conducted to validate the applicability and effectiveness of the method. The results indicated that the ‘top k%’ method can facilitate the equality of robot workload and enhance the adjacency of paths. The top k% method outperforms the existing solutions concerning a reduction of turns and start-arc points, which improves the forming quality of MRC-WAAM.

Objective To delineate the efficacy and safety profile of hemodiafiltration with endogenous reinfusion (HFR) for uremic toxin removal in patients undergoing maintenance hemodialysis (MHD).Methods Patients who have been on MHD for a period of at least 3 months were enrolled. Each subject underwent one HFR and one hemodiafiltration (HDF) treatment. Blood samples were collected before and after a single HFR or HDF treatment to test uremic toxin levels and to calculate clearance rate. The primary efficacy endpoint was to compare uremic toxin levels of indoxyl sulfate (IS), λ-free light chains (λFLC), and β2-microglobulin (β2-MG) before and after HFR treatment. Secondary efficacy endpoints was to compare the levels of urea, interleukin-6 (IL-6), P-cresol, chitinase-3-like protein 1 (YKL-40), leptin (LEP), hippuric acid (HPA), trimethylamine N-oxide (TMAO), asymmetric dimethylarginine (ADMA), tumor necrosis factor-α (TNF-α), fibroblast growth factor 23 (FGF23) before and after HFR treatment. The study also undertook a comparative analysis of uremic toxin clearance between a single HFR and HDF treatment. Meanwhile, the lever of serum albumin and branched-chain amino acids before and after a single HFR or HDF treatment were compared. In terms of safety, the study was meticulous in recording vital signs and the incidence of adverse events throughout its duration.Results The study enrolled 20 patients. After a single HFR treatment, levels of IS, λFLC, β2-MG, IL-6, P-cresol, YKL-40, LEP, HPA, TMAO, ADMA, TNF-α, and FGF23 significantly decreased (p 0.05), while post-HDF, levels of serum albumin significantly decreased (p = 0.000).Conclusion HFR treatment effectively eliminates uremic toxins from the bloodstream of patients undergoing MHD, especially protein-bound toxins and large middle-molecule toxins. Additionally, it retains essential physiological compounds like albumin and branched-chain amino acids, underscoring its commendable safety profile.

Purpose: To evaluate the potential efficacy of Triptolide (TP) on cerebral ischemia/reperfusion injury (CIRI) and to uncover the underlying mechanism through which TP regulates CIRI. Methods: We constructed a middle cerebral artery occlusion/reperfusion (MCAO/R) mouse model to simulate CIRI, and established a lipopolysaccharide (LPS)-stimulated BV-2 cell model to mimic the inflammatory state during CIRI. The neurological deficits score (NS) of mice were measured for assessment of neurologic functions. Both the severity of cerebral infarction and the apoptosis level in mouse brain tissues or cells were respectively evaluated using corresponding techniques. The expression levels of Ionized calcium binding adapter molecule 1 (IBA-1), Inductible Nitric Oxide Synthase (iNOS), Arginase 1 (Arg-1), Tumor necrosis factor-α (TNF-α), Interleukin 1β (IL-1β), Cysteine histoproteinase S (CTSS), Fractalkine, chemokine C-X3-C motif receptor 1 (CX3CR1), BCL-2-associated X protein (BAX), and antiapoptotic proteins (Bcl-2) were detected using immunofluorescence, qRT-PCR as well as Western blot, respectively. Results: Relative to the Sham group, treatment with TP attenuated the increased NS, infarct area and apoptosis levels observed in MCAO/R mice. Upregulated expression levels of IBA-1, iNOS, Arg-1, TNF-α and IL-1β were found in MCAO/R mice, while TP suppressed iNOS, TNF-α and IL-1β expression, and enhanced Arg-1 expression in both MCAO/R mice and LPS-stimulated BV-2 cells. Besides, TP inhibited the CTSS/Fractalkine/CX3CR1 pathway activation in both MCAO/R mice and LPS-induced BV-2 cells, while overexpression of CTSS reversed such effect. Co-culturing HT-22 cells with TP+LPS-treated BV-2 cells led to enhanced cell viability and decreased apoptosis levels. However, overexpression of CTSS further aggravated HT-22 cell injury. Conclusion: TP inhibits not only microglia polarization towards the M1 phenotype by suppressing the CTSS/Fractalkine/CX3CR1 pathway activation, but also HT-22 apoptosis by crosstalk with BV-2 cells, thereby ameliorating CIRI. These findings reveal a novel mechanism of TP in improving CIRI, and offer potential implications for addressing the preventive and therapeutic strategies of CIRI.

Background and Objective: Primary stability evaluation of screw implants through pull-out or push-in experiments is commonly used to investigate the mechanism of screw loosening. Numerical models simulating these testing methods could provide an enhanced understanding of the underlying attachment mechanisms as well as save time and cost in the development of new screws. However, previous numerical models have been limited by compromises between modelling the trabecular structure at high resolution versus incorporating sophisticated mechanical properties and boundary conditions, leading to overestimated mechanical performance. The aim of this study was to overcome these limitations. Methods: We developed explicit models incorporating the microstructure of trabecular bone, with frictional contact, and a non-linear material model incorporating damage. One model digitally inserted the screw into the trabecular bone structure using Boolean operations, while another model simulated the screw's rotational insertion. Results: The results showed a strong correlation between numerical and experimental results (R2: 0.54–0.93) for force-displacement response in terms of stiffness and strength. We found that the damage induced by the screw insertion process is an important factor to be considered, as the absence of modelling it led to an overestimated stiffness in previous studies. Conclusions: The study highlights the importance of including frictional contact and also identified screw insertion damage as an important part of the simulating screw-bone interaction. Our findings demonstrate the potential of explicit finite element models for accurately replicating experimental push-in results and optimizing orthopaedic screws. The code is available at https://github.com/zhou436/Bone-Screw-Constructs-eFEM.

A recombinant inbred line (RIL) population derived from wheat landrace Qingxinmai and breeding line 041133 exhibited segregation in resistance to powdery mildew and stripe rust in five and three field tests, respectively. A 16K genotyping by target sequencing (GBTS) single-nucleotide polymorphism (SNP) array-based genetic linkage map was used to dissect the quantitative trait loci (QTLs) for disease resistance. Four and seven QTLs were identified for adult-plant resistance (APR) against powdery mildew and stripe rust. QPm.caas-1B and QPm.caas-5A on chromosomes 1B and 5A were responsible for the APR against powdery mildew in line 041133. QYr.caas-1B, QYr.caas-3B, QYr.caas-4B, QYr.caas-6B.1, QYr.caas-6B.2, and QYr.caas-7B detected on the five B-genome chromosomes of line 041133 conferred its APR to stripe rust. QPm.caas-1B and QYr.caas.1B were co-localized with the pleiotropic locus Lr46/Yr29/Sr58/Pm39/Ltn2. A Kompetitive Allele Specific Polymorphic (KASP) marker KASP_1B_668028290 was developed to trace QPm/Yr.caas.1B. Four lines pyramiding six major disease resistance loci, PmQ, Yr041133, QPm/Yr.caas-1B, QPm.caas-2B.1, QYr.caas-3B, and QPm.caas-6B, were developed. They displayed effective resistance against both powdery mildew and stripe rust at the seedling and adult-plant stages.

AbstractZygophyllum brachypterum Karelin & Kirilov belongs to Zygophyllaceae and is mainly distributed in the desert regions of Central Asia, Mongolia, and Northwest China. The species is valuable in exploring the adaptations of Zygophyllaceae plants to salt stress in ecological environments. In this study, we report the complete chloroplast (cp) genome of Z. brachypterum. The entire cp genome was 104590 bp in length, with a large single-copy region (LSC, 79170 bp), a small single-copy region (SSC, 16778 bp), and two inverted repeats (IRa/IRb) of 4321 bp each. A total of 106 genes were detected, among which seven were located in the IRs, and 65, 30, and 4 were protein-coding, tRNA, and rRNA genes, respectively. Notably, eleven genes encoding the subunits of NAD(P)H dehydrogenase complex (NDH) were absent. Phylogenetic analysis indicated that Z. brachypterum belonged to Zygophylloideae (Zygophyllaceae). Furthermore, it was closely related to Z. fabago and Z. kansuense.

ABSTRACT Purpose: To investigate the protective effect of breviscapine on myocardial ischemia-reperfusion injury (MIRI) in diabetes rats. Methods: Forty rats were divided into control, diabetes, MIRI of diabetes, and treatment groups. The MIRI of diabetes model was established in the latter two groups. Then, the treatment group was treated with 100 mg/kg breviscapine by intraperitoneal injection for 14 consecutive days. Results: After treatment, compared with MIRI of diabetes group, in treatment group the serum fasting blood glucose, fasting insulin, homeostasis model assessment of insulin resistance, and glycosylated hemoglobin levels decreased, the serum total cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels decreased, the serum high-density lipoprotein cholesterol level increased, the heart rate decreased, the mean arterial pressure, left ventricular ejection fraction, and fractional shortening increased, the serum cardiac troponin I, and creatine kinase-MB levels decreased, the myocardial tumor necrosis factor α and interleukin-6 levels decreased, the myocardial superoxide dismutase level increased, and the myocardial malondialdehyde level decreased (all P < 0.05). Conclusions: For treating MIRI of diabetes in rats, the breviscapine can reduce the blood glucose and lipid levels, improve the cardiac function, reduce the myocardial injury, and decrease the inflammatory response and oxidative stress, thus exerting the alleviating effect.

3D seismic data recently acquired from the Ordos Basin shows three sets of regularly distributed fault systems, which overrides previous understanding that no faults were developed in this basin. Seismic interpretation suggests that the faults in the southwestern Ordos Basin have three basic characteristics, namely extreme micro-scale, distinct vertical stratification, and regularity of planar distribution. These NS-, NW-, and NE-trending fault systems developed in the Meso-Neoproterozoic – Lower Ordovician strata. Of these, the NS-trending fault system mainly consists of consequent and antithetic faults which show clear syndepositional deformation. The fault systems in the Carboniferous – Middle-Lower Triassic strata are not clear on seismic reflection profiles. The NW- and NE-trending fault systems are developed in the Upper Triassic – Middle Jurassic strata. Of these, the NW-trending fault system appears as a negative flower structure in sectional view and in an en echelon pattern in plan-view; they show transtensional deformation. A NE-trending fault system that developed in the Lower Cretaceous – Cenozoic strata shows a Y-shaped structural style and tension-shear properties. A comprehensive analysis of the regional stress fields at different geologic times is essential to determine the development, distribution direction, and intensity of the activity of fault systems in the Ordos Basin. Current exploration suggests three aspects in which the faults within the Ordos Basin are crucial to oil and gas accumulation. Firstly, these faults serve as vertical barriers that cause the formation of two sets of relatively independent petroleum systems in the Paleozoic and Mesozoic strata respectively; this is the basis for the ‘upper oil and lower gas’ distribution pattern. Secondly, the vertical communication of these faults is favorable for oil and gas migration, thus contributing to the typical characteristics of multiple oil and gas fields within the basin, i.e. oil and gas reservoirs with multiple superimposed strata. Finally, these faults and their associated fractures improve the permeability of Mesozoic tight reservoirs, providing favorable conditions for oil enrichment in areas around the fault systems.

Abstract The transition from deep dormancy to seed germination is essential for the life cycle of plants, but how this process occurs in the gymnosperm Chinese yew (Taxus chinensis var mairei), the natural source of the anticancer drug paclitaxel, remains unclear. Herein, we analyse the transcriptome, proteome, spatial metabolome, and spatial lipidome of the Chinese yew and present the multi-omics profiles of dormant and germinating seeds. Our results show that abscisic acid and gibberellic acid 12 homoeostasis is closely associated with gene transcription and protein translation, and the balance between these phytohormones thereby determines if seeds remain dormant or germinate. We find that an energy supply of carbohydrates from glycolysis and the TCA cycle feed into the pentose phosphate pathway during seed germination, and energy supplied from lipids are mainly derived from the lipolysis of triacylglycerols. Using mass spectrometry imaging, we demonstrate that the spatial distribution of plant hormones and phospholipids has a remarkable influence on embryo development. We also provide an atlas of the spatial distribution of paclitaxel C in Chinese yew seeds for the first time. The data from this study enable exploration of the germination mechanism of Chinese yew seeds across several omics levels.

Investigation of the dynamic properties of implants is essential to ensure safety and compatibility with the host's natural spinal tissue. This paper presents a simplified model of a cantilever beam to investigate the effects of holes/pores on the structures. Free vibration test is one of the most effective methods to measure the dynamic response of a cantilever beam, such as natural frequency and damping ratio. In this study, the natural frequencies of cantilever beams made of polycarbonate (PC) containing various circular open holes were investigated numerically, analytically, and experimentally. The experimental data confirmed the accuracy of the natural frequencies of the cantilever beam with open holes calculated by finite element and analytical models. In addition, two finite element simulation methods, the dynamic explicit and modal dynamic methods, were applied to determine the damping ratios of cantilever beams with open holes. Finite element analysis accurately simulated the damped vibration behavior of cantilever beams with open holes when known material damping properties were applied. The damping behavior of cantilever beams with random pores was simulated, highlighting a completely different relationship between porosity, natural frequency and damping response. The latter highlights the potential of finite element methods to analyze the dynamic response of arbitrary and complex structures, towards improved implant design.

The NAC family of transcription factors (TFs) is recognized as a significant group within the plant kingdom, contributing crucially to managing growth and development processes in plants, as well as to their response and adaptation to various environmental stressors. Ammopiptanthus mongolicus, a temperate evergreen shrub renowned for its remarkable resilience to low temperatures and drought stress, presents an ideal subject for investigating the potential involvement of NAC TFs in stress response mechanisms. Here, the structure, evolution, and expression profiles of NAC family TFs were analyzed systematically, and a cold and osmotic stress-induced member, AmNAC24, was selected and functionally characterized. A total of 86 NAC genes were identified in A. mongolicus, and these were divided into 15 groups. Up to 48 and 8 NAC genes were generated by segmental duplication and tandem duplication, respectively, indicating that segmental duplication is a predominant mechanism in the expansion of the NAC gene family in A. mongolicus. A considerable amount of NAC genes, including AmNAC24, exhibited upregulation in response to cold and osmotic stress. This observation is in line with the detection of numerous cis-acting elements linked to abiotic stress response in the promoters of A. mongolicus NAC genes. Subcellular localization revealed the nuclear residence of the AmNAC24 protein, coupled with demonstrable transcriptional activation activity. AmNAC24 overexpression enhanced the tolerance of cold and osmotic stresses in Arabidopsis thaliana, possibly by maintaining ROS homeostasis. The present study provided essential data for understanding the biological functions of NAC TFs in plants.

Plant cuticular wax forms a hydrophobic structure in the cuticle layer covering epidermis as the first barrier between plants and environments. Ammopiptanthus mongolicus, a leguminous desert shrub, exhibits high tolerances to multiple abiotic stress. The physiological, chemical, and transcriptomic analyses of epidermal permeability, cuticular wax metabolism and related gene expression profiles under osmotic stress in A. mongolicus leaves were performed. Physiological analyses revealed decreased leaf epidermal permeability under osmotic stress. Chemical analyses revealed saturated straight-chain alkanes as major components of leaf cuticular wax, and under osmotic stress, the contents of total wax and multiple alkane components significantly increased. Transcriptome analyses revealed the up-regulation of genes involved in biosynthesis of very-long-chain fatty acids and alkanes and wax transportation under osmotic stress. Weighted gene co-expression network analysis identified 17 modules and 6 hub genes related to wax accumulation, including 5 enzyme genes coding KCS, KCR, WAX2, FAR, and LACS, and an ABCG transporter gene. Our findings indicated that the leaf epidermal permeability of A. mongolicus decreased under osmotic stress to inhibit water loss via regulating the expression of wax-related enzyme and transporter genes, further promoting cuticular wax accumulation. This study provided new evidence for understanding the roles of cuticle lipids in abiotic stress tolerance of desert plants.

Apocynum venetum L. is a natural fiber and medicinal plant species with significant economic value. Jasmonic acid is an endogenous growth regulatory substance present in higher plants that participate in plant growth, development, and defense. As important endogenous single-stranded RNA molecules, microRNA (miRNA) plays an important role in the post-transcriptional regulation of plant genes. A combination of miRNA and mRNA sequencing techniques was used to systematically identify the methyl jasmonate miRNAs and mRNAs in A. venetum. Up to 135 conserved and 26 species-specific miRNAs have been identified in A. venetum. These miRNAs mainly target genes that encode transcription factors and enzymes. The expression levels of 23 miRNAs, including miR398 and miR482, significantly changed after MeJA treatment. A total of 1778 genes were differentially expressed under MeJA treatment, of which 825 were upregulated and 953 were downregulated. The main biological processes enriched in these differentially expressed genes were redox balance, secondary metabolism, photosynthesis, and plant hormone signal transduction. Joint analysis of the miRNAs and mRNA revealed that MeJA-responsive miRNAs function by forming regulatory modules, including miR398-CSD, miR482-NBS-LRR, miR156-SPL10, and miR164-NAC056, which further regulate multiple biological processes, including redox balance, disease resistance, and morphogenesis in A. venetum. This study provides important information to understand the biological roles of miRNAs in A. venetum.

Background Diagnosis of asthma and chronic obstructive pulmonary disease (COPD) in the community is variable, often without spirometry. Some studies report that adults with both diagnostic labels (asthma+COPD) have worse health outcomes than those with asthma or COPD only, but data for Australian adults are limited. We investigated the relationship between clinical characteristics and self-reported diagnoses of asthma, COPD and both. Method We used data from the BOLD Australia study, which included randomly selected adults aged ≥40 years from six study sites. The BOLD questionnaires and spirometry test were used in all sites. Participants were grouped by self-reported diagnosis. Demographic and clinical characteristics and lung function were compared between groups. Results Of the study sample (n=3522), 336 reported asthma only, 172 reported COPD only, 77 reported asthma+COPD and 2937 reported neither. Fewer than half of participants with a COPD diagnosis (with or without asthma) had airflow limitation. Participants with asthma+COPD had more respiratory symptoms and greater airflow limitation than those with either diagnosis alone. Having asthma+COPD was independently associated with a higher probability of having clinically important breathlessness (modified Medical Research Council score ≥2) than asthma only (adjusted OR 3.44, 95% CI 1.86–6.33) or COPD only (adjusted OR 3.28, 95% CI 1.69–6.39). Airflow limitation (Global Initiative for Chronic Obstructive Lung Disease 2 or higher, using post-bronchodilator forced expiratory volume in 1 s/forced vital capacity ratio

Thermopsis lanceolata R. Br. belongs to the genus Thermopsis, Fabaceae. The alkaloids of T. lanceolata have anti-cancer, anti-heart rate disorders and other pharmacological effects. To explore the chloroplast genome of T. lanceolata and its phylogenetic relationship, a complete chloroplast genome of T. lanceolata was sequenced and annotated, and a phylogenetic tree was constructed. The complete chloroplast genome of T. lanceolata is a circular molecule of 151,526 bp, consisting of a large single copy (LSC) region of 83,780 bp, a small single copy (SSC) region of 15,566 bp, and a pair of inverted repeats (IRa and IRb) of 26,090 bp. The chloroplast genome of T. lanceolata contained 130 genes, including 85 protein-coding genes, 37 transfer RNA genes, and 8 ribosomal RNA genes. Phylogenetic analysis revealed a close relationship between T. lanceolata and T. turkestanica.

Abstract Background Observational studies have shown home hemodialysis (HHD) to be associated with better survival than facility hemodialysis (HD) and peritoneal dialysis (PD). Patients on HHD have reported higher quality of life and independence. HHD is considered to be an economical way to manage end-stage kidney disease (ESKD). The coronavirus disease 2019 pandemic has had a significant impact on patients with ESKD. Patients on HHD may have an advantage over in-center HD patients because of a lower risk of exposure to infection. Participants and methods We enrolled HD patients from our dialysis center. We first established the HHD training center. The training center was approved by the Chinese government. Doctors, nurses and engineers train and assess patients separately. There are three forms of patient monitoring: home visits, internet remote monitoring, and outpatient services. Demographic and medical data included age, sex, blood pressure, and dialysis-related data. Laboratory tests were conducted in our central testing laboratory, including hemoglobin (Hgb), serum creatinine (Cr), urea nitrogen (BUN), uric acid (UA), albumin (Alb), calcium (Ca), phosphorus (P), parathyroid hormone (PTH), and brain natriuretic peptide (BNP) levels. Results Six patients who underwent regular dialysis in the HD center of our hospital were selected for HHD training. We enrolled 6 patients, including 4 males and 2 females. The mean age of the patients was 47.5 (34.7-55.7) years, and the mean dialysis age was 33.5 (11.2-41.5) months. After an average of 16.0 (11.2-25.5) months of training, Alb, P and BNP levels were improved compared with the baseline values. After training, three patients returned home to begin independent HD. During the follow-up, there were no serious adverse events leading to hospitalization or death, but there were several adverse events. They were solved quickly by extra home visits of the technicians or online by remote monitoring. During the follow-up time, the laboratory indicators of all the patients, including Hgb, Alb, Ca, P, PTH, BNP, and β2-MG levels, remained stable before and after HHD treatment. Conclusion HHD is feasible and safe for ESKD in China, but larger-scale and longer-term studies are needed for further confirmation.

ABSTRACT Purifying extracellular vesicles (EVs) has been challenging because EVs are heterogeneous in cargo yet share similar sizes and densities. Most surface marker‐based affinity separation methods are limited to research or diagnostic scales. We report that heparin chromatography can separate purified EVs into two distinct subpopulations as ascertained by MS/MS: a non‐heparin‐binding (NHB) fraction that contains classical EV markers such as tetraspanins and a heparin‐binding (HB) fraction enriched in fibronectins and histones. Both fractions were similarly fusogenic but induced different transcriptional responses in endothelial cells. While EVs that were purified by conventional, non‐affinity methods alone induced ERK1/2 phosphorylation and Ki67, the NHB fraction did not. This result suggests heparin chromatography as an additional novel fractionation step that is inherently scalable, does not lead to loss of material, and separates inflammatory and pyrogenic EVs from unreactive EVs, which will improve clinical applications.

IntroductionHyperbaric oxygen therapy (HBOT) is one of the common clinical treatments, but adverse effects have hampered and limited the clinical application and promotion of hyperbaric oxygen therapy. A systematic review and meta-analysis of the adverse effects of hyperbaric oxygen therapy have conducted by our group to provide a theoretical basis for clinical treatment.MethodsThree electronic databases (PubMed, Web of Science, and The Cochrane Library) were comprehensively searched for randomized clinical trials (RCTs) from March 2012 to October 2022. Two reviewers independently screened titles and abstracts for eligibility and assessed the quality of the included studies. The meta-analysis was performed using RevMan 5.3.ResultsA total of 24 RCTs involving 1,497 participants were identified. ① The HBOT group reported more adverse effects (30.11% vs. 10.43%, p 10 sessions, the incidence of adverse effects was higher than that of the control group; when the course of HBOT was ≤10 sessions, the adverse effects caused by hyperbaric oxygen were comparatively lower. ④ When the chamber pressure is above 2.0 ATA, the incidence of adverse effects is higher than that of the control group. While the chamber pressure is lower than 2.0 ATA, HBOT is relatively safe compared with the previous one.ConclusionHyperbaric oxygen therapy (HBOT) is more likely to cause adverse reactions when the chamber pressure is above 2.0 ATA. More attention should be paid to the possible occurrence of related adverse effects if the treatment course is >10 sessions.Systematic review registrationhttps://www.crd.york.ac.uk/PROSPERO/, identifier CRD42022316605.

R2R3-MYB transcription factors (TFs) participate in the modulation of plant development, secondary metabolism, and responses to environmental stresses. Ammopiptanthus nanus, a leguminous dryland shrub, tolerates a high degree of environmental stress, including drought and low-temperature stress. The systematic identification, structural analysis, evolutionary analysis, and gene profiling of R2R3-MYB TFs under cold and osmotic stress in A. nanus were performed. Up to 137 R2R3-MYB TFs were identified and clustered into nine clades, with most A. nanus R2R3-MYB members belonging to clade VIII. Tandem and segmental duplication events drove the expansion of the A. nanus R2R3-MYB family. Expression profiling revealed that multiple R2R3-MYB genes significantly changed under osmotic and cold stress conditions. MiR858 and miR159 targeted 88 R2R3-MYB genes. AnaMYB87, an miR858-targeted clade VIII R2R3-MYB TF, was up-regulated under both osmotic and cold stress. A transient expression assay in apples showed that the overexpression of AnaMYB87 promoted anthocyanin accumulation. A luciferase reporter assay in tobacco demonstrated that AnaMYB87 positively affected the transactivation of the dihydroflavonol reductase gene, indicating that the miR858-MYB87 module mediates anthocyanin accumulation under osmotic stress by regulating the dihydroflavonol reductase gene in A. nanus. This study provides new data to understand the roles of R2R3-MYB in plant stress responses.

Ammopiptanthus mongolicus, a traditional ethnic medicinal herb, is a rare broad-leaved evergreen shrub in the arid region of central Asia that can survive under extremely low temperatures during winter. In this study, we investigated the cold stress response of A. mongolicus leaves using physiological and proteomic approaches. Cold stress treatment increased the relative electrolyte leakage, proline, and soluble sugar levels and decreased the chlorophyll content in A. mongolicus leaves. Moreover, 93 differentially abundant proteins (DAPs) were identified using two-dimensional electrophoresis, of which 79 were further confirmed via tandem mass spectrometric analysis. The predicted functions of DAPs were mainly associated with photosynthesis in chloroplasts, reactive oxygen species scavenging, defense, and protein synthesis, folding, and degradation. A. mongolicus chitinase A (AmCHIA) is a cold-induced apoplast protein whose transcription is upregulated under cold, osmotic, high-salinity, and mechanical stresses. Recombinant AmCHIA expressed in Escherichia coli exhibits chitinase activity. Here, AmCHIA expression enhanced the cold tolerance of E. coli cells, suggesting that it may contribute to the cold adaptation of A. mongolicus after cold treatment. The present study not only provides important data for understanding the cold stress responses in plants but also serves as a basis for further exploration of the biological functions of cold-induced proteins in A. mongolicus using genetic approaches.

Accurate phenotypic identification is the basis of research for resistance genetics and rice breeding for resistance to RBSDV disease. Obtaining rice black-streaked dwarf virus (RBSDV) viruliferous small brown planthoppers (SBPHs) with high transmission efficiency is an essential part of accurate phenotypic identification. Here, through quantifying number of RBSDV copies in infected rice plants, optimizing times of SBPHs fed on RBSDV-infected rice plants and leaf stage of rice seedlings, a method to acquire an RBSDV-carrying SBPH population more efficiently was improved. The results showed that rate of viruliferous SBPHs was significantly higher when fed on RBSDV-infected rice plants that had the copy numbers of RBSDV S10 of 3.0*104 and 1.1*104 than 8.3*102. Therefore, it is more efficient for SBPHs to acquire the virus when fed on RBSDV-infected rice plants that have copy numbers of RBSDV S10 above 1.1*104. The rate of viruliferous SBPHs were 50% and 54%, respectively, after the insects fed on RBSDV-infected rice plants for 7 and 9 days and being transferred to healthy rice seedlings for 5 and 3 days, which was significantly higher than those at other feeding times. The optimal inoculation leaf stage of rice seedlings was the 2-3-leaf stage (3 effective SBPHs per seedling for 72 h), but a high rate of viruliferous SBPHs may be suggested for inoculation of older rice seedlings.

Abstract Background Understanding the host genetic architecture and viral immunity contributes to the development of effective vaccines and therapeutics for controlling the COVID-19 pandemic. Alterations of immune responses in peripheral blood mononuclear cells play a crucial role in the detrimental progression of COVID-19. However, the effects of host genetic factors on immune responses for severe COVID-19 remain largely unknown. Methods We constructed a computational framework to characterize the host genetics that influence immune cell subpopulations for severe COVID-19 by integrating GWAS summary statistics (N = 969,689 samples) with four independent scRNA-seq datasets containing healthy controls and patients with mild, moderate, and severe symptom (N = 606,534 cells). We collected 10 predefined gene sets including inflammatory and cytokine genes to calculate cell state score for evaluating the immunological features of individual immune cells. Results We found that 34 risk genes were significantly associated with severe COVID-19, and the number of highly expressed genes increased with the severity of COVID-19. Three cell subtypes that are CD16+monocytes, megakaryocytes, and memory CD8+T cells were significantly enriched by COVID-19-related genetic association signals. Notably, three causal risk genes of CCR1, CXCR6, and ABO were highly expressed in these three cell types, respectively. CCR1 +CD16+monocytes and ABO + megakaryocytes with significantly up-regulated genes, including S100A12, S100A8, S100A9, and IFITM1, confer higher risk to the dysregulated immune response among severe patients. CXCR6 + memory CD8+ T cells exhibit a notable polyfunctionality including elevation of proliferation, migration, and chemotaxis. Moreover, we observed an increase in cell-cell interactions of both CCR1 + CD16+monocytes and CXCR6 + memory CD8+T cells in severe patients compared to normal controls among both PBMCs and lung tissues. The enhanced interactions of CXCR6 + memory CD8+T cells with epithelial cells facilitate the recruitment of this specific population of T cells to airways, promoting CD8+T cell-mediated immunity against COVID-19 infection. Conclusions We uncover a major genetics-modulated immunological shift between mild and severe infection, including an elevated expression of genetics-risk genes, increase in inflammatory cytokines, and of functional immune cell subsets aggravating disease severity, which provides novel insights into parsing the host genetic determinants that influence peripheral immune cells in severe COVID-19.

Abstract Background Primary biliary cholangitis (PBC) is a classical autoimmune disease, which is highly influenced by genetic determinants. Many genome-wide association studies (GWAS) have reported that numerous genetic loci were significantly associated with PBC susceptibility. However, the effects of genetic determinants on liver cells and its immune microenvironment for PBC remain unclear. Results We constructed a powerful computational framework to integrate GWAS summary statistics with scRNA-seq data to uncover genetics-modulated liver cell subpopulations for PBC. Based on our multi-omics integrative analysis, 29 risk genes including ORMDL3, GSNK2B, and DDAH2 were significantly associated with PBC susceptibility. By combining GWAS summary statistics with scRNA-seq data, we found that cholangiocytes exhibited a notable enrichment by PBC-related genetic association signals (Permuted P

Abstract Background Infiltrating immune and stromal cells are important components of the endometrial cancer (EC) microenvironment, which has a significant effect on the biological behavior of EC, suggesting that unique immune-related genes may be associated with the prognosis of EC. However, the association of immune-related genes with the prognosis of EC has not been elucidated. We attempted to identify immune-related genes with potentially prognostic value in EC using The Cancer Genome Atlas database and the relationship between immune microenvironment and EC. Methods We analyzed 578 EC samples from TCGA database and used weighted gene co-expression network analysis to screen out immune-related genes. We constructed a protein–protein interaction network and analyzed it using STRING and Cytoscape. Immune-related genes were analyzed through conjoint Cox regression and random forest algorithm analysis were to identify a multi-gene prediction model and stratify low-risk and high-risk groups of EC patients. Based on these data, we constructed a nomogram prediction model to improve prognosis assessment. Evaluation of Immunological, gene mutations and gene enrichment analysis were applied on these groups to quantify additional differences. Results Using conjoint Cox regression and random forest algorithm, we found that TRBC2, TRAC, LPXN, and ARHGAP30 were associated with the prognosis of EC and constructed four gene risk models for overall survival and a consistent nomogram. The time-dependent receiver operating characteristic curve analysis revealed that the area under the curve for 1-, 3-, and 5-y overall survival was 0.687, 0.699, and 0.76, respectively. These results were validated using a validation cohort. Immune-related pathways were mostly enriched in the low-risk group, which had higher levels of immune infiltration and immune status. Conclusion Our study provides new insights for novel biomarkers and immunotherapy targets in EC.

Abstract Background Anemia is one of the main complications of chronic kidney disease especially kidney failure, which includes treatment with erythropoiesis-stimulating agents and iron supplementation, including intravenous and oral iron. However, intravenous iron may pose limitations, such as potential infusion reactions. Oral iron is mainly composed of divalent iron, which can excessively stimulate the gastrointestinal tract. Iron polysaccharide complex capsules are a novel oral iron trivalent supplement with higher iron content and lower gastrointestinal irritation. However, since high-quality evidence-based medicinal support is lacking, it is necessary to conduct clinical studies to further evaluate the effectiveness and safety of oral iron polysaccharide complex in chronic kidney disease patients. Methods This randomized controlled trial uses an open-label, parallel group design, where the efficacy and safety of maintenance hemodialysis (MHD) participants is evaluated. The experimental group is assigned erythropoietins and iron polysaccharide complex (two capsules each time, bid), and the control group is assigned erythropoietin and sucrose iron (100mg, 2w) injection. Participants (aged 18–75 years) undergoing maintenance hemodialysis were considered for screening. Inclusion criteria included hemoglobin (Hb) ≥110g/L and < 130g/L, transferrin saturation (TSAT) > 20% and < 50%, and serum ferritin (SF) > 200μg/L and < 500μg/L. Exclusion criteria included acute or chronic bleeding, serum albumin < 35g/L, hypersensitive C-reactive protein (HsCRP) > 10 mg/L, and severe secondary hyperparathyroidism (iPTH ≥ 800 pg/mL). Full inclusion and exclusion criteria are described in the “Methods” section. The primary endpoint is TSAT of the participants at week 12. Secondary endpoints include Hb, SF, hematocrit (Hct), HsCRP, pharmacoeconomic evaluation, drug costs, quality of life, and indicators of oxidative stress. The treatment will last for 24 weeks with a follow-up visit at baseline (within 7 days prior to initial treatment) and weeks 4, 8, 12, 16, 20, and 24 after initial treatment. This clinical research includes 9 hemodialysis centers in mainland China and plans to enroll 186 participants. Discussion It is expected that it will provide strong evidence to reveal the clinical efficacy and safety of oral iron in the treatment of chronic CKD-related anemia in MHD patients through this clinical trial. Trial registration Chinese Clinical Trial Registry ChiCTR2000031166 . Registered on March 23, 2020

BackgroundCongenital pseudarthrosis of the tibia (CPT) is a refractory and rare disease. Because of its extremely low incidence, little is known about its clinical features. In this retrospective study, we aim to analyze the clinical characteristics of patients with CPT.Materials and methodsThis is a retrospective study of children with CPT identified by the radiological review. Investigations of CPT included general conditions, the characteristics of CPT, treatment methods, and surgical complications.ResultsWe collected 514 CPT cases from March 1999 to March 2020 in our hospital, such as 317 (61.67%) boys, 197 (38.33%) girls; 330 (62.86%) in Crawford IV; 510 (97.14%) in mid and distal 1/3 tibia; 481 (93.58%) in less than 3 years at onset age; 297 (57.78%) in less than 3 years at the first outpatient visit. The most common post-operative complication was ankle valgus (101, 39.60%), followed by limb length discrepancy (91, 35.69%), refracture (38, 14.90%), osteomyelitis (15, 5.88%), and removal of internal fixation (10, 3.93%).ConclusionsCPT with a higher incidence of Crawford IV frequently occurs in boys and the middle or distal part of the tibia; most patients have the onset age and first outpatient visit before 3 years; the major surgical complications are ankle valgus and limb length discrepancy.

Summary: Background: White matter (WM) microstructural abnormalities have been observed in diabetes. However, evidence of prediabetes is currently lacking. This study aims to investigate the WM integrity in prediabetes and diabetes. We also assess the association of WM abnormalities with glucose metabolism status and continuous glucose measures. Methods: The WM integrity was analyzed using cross-sectional baseline data from a population-based PolyvasculaR Evaluation for Cognitive Impairment and vaScular Events (PRECISE) study. The cohort, including a total of 2218 cases with the mean age of 61.3 ± 6.6 years and 54.1% female, consisted of 1205 prediabetes which are categorized into two subgroups (a group of 254 prediabetes with combined impaired fasting glucose (IFG) and impaired glucose tolerance (IGT) and the other group of 951 prediabetes without combined IFG/IGT), 504 diabetes, and 509 normal control subjects. Alterations of WM integrity were determined by diffusion tensor imaging along with tract-based spatial statistics analysis to compare diffusion metrics on WM skeletons between groups. The mixed-effects multivariate linear regression models were used to assess the association between WM microstructural alterations and glucose status. Findings: Microstructural abnormalities distributed in local WM tracts in prediabetes with combined IFG/IGT and spread widely in diabetes. These WM abnormalities are associated with higher glucose measures. Interpretation: Our findings suggest that WM microstructural abnormalities are already present at the prediabetes with combined IFG/IGT stage. Preventative strategies should begin early to maintain normal glucose metabolism and avert further destruction of WM integrity. Funding: Partially supported by National Key R&D Program of China (2016YFC0901002).

As a new support structure system, the double-row pile is widely used in foundations in pit engineering, slope engineering, water conservancy engineering, and other fields. It has a big horizontal stiffness and strong anti-overturning ability and has less impact on the surrounding neighboring existing buildings (structures) when it is applied. It must be mentioned that the design experience of the double-row pile is ahead of the theory, and some research has been carried out on double-row pile support structures. This work reviews and evaluates the force deformation and earth pressure on piles of double-row piles, including pile deformation damage mechanism, soil pressure distribution law, and double-row pile structure form. In addition, the problems and shortcomings of the current double-row pile research are concluded. These results have enriched the study of the mechanism of double-row piles, provided the theoretical guidance for double-row pile support structures, and laid the foundation for further theoretical research.

The response of soil microorganisms to altered nitrogen (N) and rainfall patterns plays an important role in understanding ecosystem carbon and nitrogen cycling processes under global change. Previous studies have separately focused on the effects of N addition and rainfall on soil microbial diversity and community composition. However, the combined and interactive impact of N addition and rainfall on soil microbial diversity and function mediated by plant and soil processes have been poorly investigated for grassland ecosystems. Here, we conducted a field experiment with simulated N addition (N addition: 10 g N m–2yr–1) and altered rainfall pattern [control, rainfall reduction (compared to control –50%); rainfall addition (compared to control + 50%)] to study their interactive effects on soil microbial diversity and function in a temperate steppe of Inner Mongolia. Our results showed that N addition and rainfall addition significantly increased soil bacterial diversity, and the bacterial diversity was positively correlated with soil microbial biomass nitrogen, inorganic nitrogen, and Stipa krylovii root exudate C:N ratio, Allium polyrhizum root exudate C and N, and A. polyrhizum root exudate C:N ratio. N addition and rainfall reduction significantly reduced fungal diversity, which correlated closely with soil microbial biomass carbon and the C:N ratio of A. polyrhizum root exudates. Bacteria were mainly eutrophic r-strategists, and the responses of bacterial function guilds to the interaction between N addition and rainfall pattern were not significant. However, the arbuscular mycorrhizal fungi (AMF), in the functional classification of fungi, were significantly reduced under the condition of N addition and rainfall reduction, and the absolute abundance of the phylum Glomeromycota increased under rainfall addition, suggesting that AMFs are sensitive to altered N and rainfall patterns over short timescales (1 year). Collectively, our results have important implications for understanding the plant–soil–microbe system of grasslands under climate change.

MicroRNAs (miRNAs) are small endogenous single-stranded RNAs that regulate plant growth, development, and environmental stress response posttranscriptionally. Ammopiptanthus nanus, a rare evergreen broad-leaved shrub in the temperate area of Central Asia, can tolerate freezing stress as low as –30 degrees centigrade in winter, and miRNA might be involved in the cold acclimation which enables A. nanus to obtain tolerance to freezing stress. Systematic identification and functional analysis of the miRNAs involved in the cold acclimation in A. nanus may promote understanding of the miRNA-mediated gene regulation network underlying cold acclimation. Here, based on small RNA and degradome sequencing, 256 miRNAs and 1,808 miRNA-target pairs were identified in A. nanus. A total of 39 cold-responsive miRNAs were identified, of which 29 were upregulated and ten were downregulated. These cold-responsive miRNAs may participate in the cold acclimation by regulating redox homeostasis (miR398, miR4415, and miR408), calcium signaling (miR5225 and miR5211), growth and development (miR159 and miR390), and small RNA–mediated gene silencing (miR168 and miR1515). We found that miR4415, a legume-specific miRNA, is involved in the cold acclimation of A. nanus by targeting an L-ascorbate oxidase gene and then regulating the redox state of the apoplast. Our study provides important data for understanding the regulatory role of miRNA in the cold acclimation of A. nanus.

Objective This study aimed to explore the effectiveness of thiamin and folic acid supplementation on the improvement of the cognitive function in patients with maintenance hemodialysis. Method In the present study, we randomly assigned patients undergoing hemodialysis who had the Montreal Cognitive Assessment (MoCA) score lower than 26 to treatment group (n = 25, thiamin 90 mg/day combined with folic acid 30 mg/day) or control group (n = 25, nonintervention). All subjects were followed up for 96 weeks. The primary outcome was the improvement of the MoCA score. The secondary outcomes included homocysteine level, survival and safety. Results Patients in treatment group had an increase of the MoCA score from 21.95 ± 3.81 at baseline to 25.68 ± 1.96 at week 96 (p

Long non-coding RNAs (lncRNAs) have been shown to play critical regulatory roles in plants. Ammopiptanthus nanus can survive under severe low-temperature stress, and lncRNAs may play crucial roles in the gene regulation network underlying the cold stress response in A. nanus. To investigate the roles of lncRNAs in the cold stress response of A. nanus, a combined lncRNA and mRNA expression profiling under cold stress was conducted. Up to 4890 novel lncRNAs were identified in A. nanus and 1322 of them were differentially expressed under cold stress, including 543 up-regulated and 779 down-regulated lncRNAs. A total of 421 lncRNAs were found to participate in the cold stress response by forming lncRNA–mRNA modules and regulating the genes encoding the stress-related transcription factors and enzymes in a cis-acting manner. We found that 31 lncRNAs acting as miRNA precursors and 8 lncRNAs acting as endogenous competitive targets of miRNAs participated in the cold stress response by forming lncRNA–miRNA–mRNA regulatory modules. In particular, a cold stress-responsive lncRNA, TCONS00065739, which was experimentally proven to be an endogenous competitive target of miR530, contributed to the cold stress adaptation by regulating TZP in A. nanus. These results provide new data for understanding the biological roles of lncRNAs in response to cold stress in plants.

The uplift of the Qinghai Tibet Plateau has led to a drastic change in the climate in Central Asia, from warm and rainy, to dry and less rainfall. Ammopiptanthus nanus, a rare evergreen broad-leaved shrub distributed in the temperate desert region of Central Asia, has survived the drastic climate change in Central Asia caused by the uplift of the Qinghai-Tibet Plateau. Ascorbate oxidase (AO) regulates the redox status of the apoplast by catalyzing the oxidation of ascorbate acid to dehydroascorbic acid, and plays a key role in the adaptation of plants to environmental changes. Analyzing the evolution, environmental response, and biological functions of the AO family of A. nanus is helpful for understanding how plant genome evolution responds to climate change in Central Asia. A total of 16 AOs were identified in A. nanus, all of which contained the ascorbate oxidase domain, most of which contained transmembrane domain, and many were predicted to be localized in the apoplast. Segmental duplication and tandem duplication are the main factors driving the gene amplification of the AO gene family in A. nanus. Gene expression analysis based on transcriptome data and fluorescence quantitative PCR, as well as enzyme activity measurements, showed that the expression levels of AO genes and total enzyme activity decreased under short-term osmotic stress and low-temperature stress, but the expression of some AO genes (AnAO5, AnAO13, and AnAO16) and total enzyme activity increased under 7 days of cold stress. AnAO5 and AnAO11 are targeted by miR4415. Further functional studies on AnAO5 showed that AnAO5 protein was localized in the apoplast. The expression of AnAO5 in yeast cells and the transient expression in tobacco enhanced the tolerance of yeast and tobacco to low-temperature stress, and the overexpression of AnAO5 enhanced the tolerance of Arabidopsis seedlings to cold stress. Our research provides important data for understanding the role of AOs in plant adaptation to environmental change.

Thaumatin-like proteins (TLPs), a family of proteins with high sequence similarity to thaumatin, are shown to be involved in plant defense, and are thus classified into the pathogenesis related protein family 5. Ammopiptanthus nanus is a rare evergreen broad-leaved shrub distributed in the temperate zone of Central Asia, which has a high tolerance to low-temperature stress. To characterize A. nanus TLPs and understand their roles in low-temperature response in A. nanus, a comprehensive analysis of the structure, evolution, and expression of TLP family proteins was performed. A total of 31 TLP genes were detected in the A. nanus genome, and they were divided into four groups based on their phylogenetic positions. The majority of the AnTLPs contained the conserved cysteine residues and were predicted to have the typical three-dimensional structure of plant TLPs. The primary modes of gene duplication of the AnTLP family genes were segmental duplication. The promoter regions of most AnTLP genes contain multiple cis-acting elements related to environmental stress response. Gene expression analysis based on transcriptome data and fluorescence quantitative PCR analysis revealed that several AnTLP genes were involved in cold-stress response. We further showed that a cold-induced AnTLP gene, AnTLP13, was localized in apoplast, and heterologous expression of the AnTLP13 in Escherichia coli and yeast cells and tobacco leaves enhanced low-temperature stress tolerance when compared with the control cells or seedlings. Our study provided important data for understanding the roles of TLPs in plant response to abiotic stress.

Abstract Extracellular vesicles (EVs) are secreted from all cell types and are intimately involved in tissue homeostasis. They are being explored as vaccine and gene therapy platforms, as well as potential biomarkers. As their size is below the diffraction limit of light microscopy, direct visualizations have been daunting and single‐particle studies under physiological conditions have been hampered. Here, direct stochastic optical reconstruction microscopy (dSTORM) was employed to visualize EVs in three‐dimensions and to localize molecule clusters such as the tetraspanins CD81 and CD9 on the surface of individual EVs. These studies demonstrate the existence of membrane microdomains on EVs. These were confirmed by Cryo‐EM. Individual particle visualization provided insights into the heterogeneity, structure, and complexity of EVs not previously appreciated

Drug-resistant epilepsy remains a significant clinical and societal burden, with one third of people with epilepsy continuing to experience seizures despite the availability of around 30 anti-seizure drugs (ASDs). Further, ASDs often have substantial adverse effects, including impacts on learning and memory. Therefore, it is important to develop new ASDs, which may be more potent or better tolerated. Here, we report the preliminary preclinical evaluation of BICS01, a synthetic product based on a natural compound, as a potential ASD. To model seizure-like activity in vitro, we prepared hippocampal slices from adult male Sprague Dawley rats, and elicited epileptiform bursting using high extracellular potassium. BICS01 (200 μM) rapidly and reversibly reduced the frequency of epileptiform bursting but did not change broad measures of network excitability or affect short-term synaptic facilitation. BICS01 was well tolerated following systemic injection at up to 1,000 mg/kg. However, we did not observe any protective effect of systemic BICS01 injection against acute seizures evoked by pentylenetetrazol. These results indicate that BICS01 is able to acutely reduce epileptiform activity in hippocampal networks. Further preclinical development studies to enhance pharmacokinetics and accumulation in the brain, as well as studies to understand the mechanism of action, are now required.

Tetraena mongolica, Zygophyllum xanthoxylon, and Z. fabago are three typical dryland plants with important ecological values in subfamily Zygophylloideae of Zygophyllaceae. Studies on the chloroplast genomes of them are favorable for understanding the diversity and phylogeny of Zygophyllaceae. Here, we sequenced and assembled the whole chloroplast genomes of T. mongolica, Z. xanthoxylon, and Z. fabago, and performed comparative genomic and phylogenetic analysis. The total size, structure, gene content and orders of these three chloroplast genomes were similar, and the three chloroplast genomes exhibited a typical quadripartite structure with a large single-copy region (LSC; 79,696-80,291 bp), a small single-copy region (SSC; 16,462-17,162 bp), and two inverted repeats (IRs; 4,288-4,413 bp). A total of 107 unique genes were identified from the three chloroplast genomes, including 70 protein-coding genes, 33 tRNAs, and 4 rRNAs. Compared with other angiosperms, the three chloroplast genomes were significantly reduced in overall length due to an unusual 16-24 kb shrinkage of IR regions and loss of the 11 genes which encoded subunits of NADH dehydrogenase. Genome-wide comparisons revealed similarities and variations between the three species and others. Phylogenetic analysis based on the three chloroplast genomes supported the opinion that Zygophyllaceae belonged to Zygophyllales in Fabids, and Z. xanthoxylon and Z. fabago belonged to Zygophyllum. The genome-wide comparisons revealed the similarity and variations between the chloroplast genomes of the three Zygophylloideae species and other plant species. This study provides a valuable molecular biology evidence for further studies of phylogenetic status of Zygophyllaceae.

The double-row pile supporting structure has been widely used in foundation pit excavations. When analyzing the effect of earth pressure on the pile structure, previous research only considered the double-row piles as the rigid body and the pile-soil interaction has not been examined. In this study, a theoretical model was developed based on Duncan-Chang’s hyperbolic theory to calculate earth pressures in the active and passive zones of the double-row pile supporting structure. The model considered the nonlinear effect of the pile deformation on the active and passive earth pressures. The macroscopic pile-soil interaction was converted into a microscopic stress-strain relationship at a certain point in the soil body, reflecting the nonlinear effect of pile deformation on earth pressure. Numerical simulation and large-scale field tests have been conducted to verify the proposed model. The results show that the average values of the parameters obtained by numerical simulation are a¯=0.38, b¯=−0.253 for the active zone and a¯=0.00612, b¯=−0.729 for the passive zone. Based on the values of a¯ and b¯, the predicted active and passive earth pressures stemming from the developed model agreed well with those obtained from field tests. The developed model in this study can be used to predict the distribution of active and passive earth pressures for double-row pile supporting structures.

This study solidifies the aeolian sand by microbial-induced carbonate precipitation (MICP) technique. The effects of cementation solution with different concentrations, particle size, and grouting batches are examined via the bender element, unconfined compressive test, and scanning electron microscope (SEM). The bender element results show that the wave speed of loose aeolian sand is 200 m/s; however, after solidification of the aeolian sand, the speed of P-wave is about 450-600 m/s and S-wave is about 350-500 m/s. Additionally, the unconfined compressive strength (UCS) results indicate that when the concentration of cementation solution is 0.75 mol/L, the UCS of biosolidified sand sample is the highest. Then, compared with the aeolian sand with original grade, the particles ranging from 0.1 to 0.4 mm have a better cementation effect. Moreover, the UCS of biosolidified sand samples increases along with the grouting batch. From the SEM images, it can be seen that when the grouting batch reaches to five times, the particles are almost completely covered by CaCO3 crystals compared with the three batches and four batches.

Abstract Background Rice black-streaked dwarf virus (RBSDV) causes one of the most important rice virus diseases of plants in East Asia. However, molecular mechanism(s)controlling rice resistance to infection is largely unknown. Results In this paper, we showed that RBSDV infection in rice significantly induced nitric oxide (NO) production. This finding was further validated through a genetic approach using a RBSDV susceptible (Nipponbare) and a RBSDV resistant (15HPO187) cultivar. The production of endogenous NO was muchhigher in the 15HPO187 plants, leading to a much lower RBSDV disease incidence. Pharmacological studies showed that the applications of NO-releasingcompounds (i.e., sodium nitroprusside [SNP] and nitrosoglutathione [GSNO]) to rice plants reduced RBSDV disease incidence. After RBSDV infection, the levels of OsICS1, OsPR1b and OsWRKY 45 transcripts were significantly up-regulated by NO in Nipponbare. The increased salicylic acid contents were also observed. After the SNP treatment, protein S-nitrosylation in rice plants was also increased, suggesting that the NO-triggered resistance to RBSDV infection was partially mediated at the post-translational level. Although Osnia2 mutant rice produced less endogenous NO after RBSDV inoculation and showed a higher RBSDV disease incidence, its RBSDV susceptibility could be reduced by SNP treatment. Conclusions Collectively, our genetic and molecular evidence revealed that endogenous NO was a vital signal responsible for rice resistance to RBSDV infection.

Abstract Rice stripe virus (RSV) causes one of the most important rice virus diseases of plants in East Asia. However, the molecular mechanisms controlling rice resistance to RSV infection are largely unknown. Recently, several studies presented a novel model that melatonin (MT) and nitric oxide (NO) participate in the plant-pathogen interaction in a synergetic manner. In this study, there was a difference in MT content between two rice varieties that correlated with one being susceptible and one being resistant to RSV, which suggested that MT is related to RSV resistance. In addition, a test with two NO biosynthesis inhibitors revealed that NO inhibitor were able to increase the disease incidence of RSV. A pharmacological experiment with exogenous MT and NO showed that increased MT and NO in the MT-pretreated plants led to lower disease incidences; however, only NO increased in a NO-releasing reagent [sodium nitroprusside (SNP)] pretreated plants. The expressions level of OsPR1b and OsWRKY 45 were significantly induced by MT and NO. These results suggest that rice resistance to RSV can be improved by increased MT through a NO-dependent pathway.

In this study, UPLC-MS/MS was used to study the effects of smoking duration and temperature on the formation of heterocyclic amines (HAs) in smoke-processed meat patties. Four kinds of free HAs—including F-7,8-DiMeIQx; F-MeAαC; F-Harman and F-Norharman—and six kinds of protein-bound HAs—including B-AαC; B-7,8-DiMeIQx; B-Glu-p-1; B-MeAαC; B-Harman and B-Norharman—were detected and quantified. Among the free HAs, we observed a 23-fold content increase (p < 0.05), from 0–4 h (at 0 h and 4 h they were 4.24 ng·g−1 and 98.33 ng·g−1, respectively), and the content of the free HAs decreased to 78.80 ng·g−1, at 5 h. At the same time, the free HAs content increased from 53.52 ng·g−1, at 50 °C, to 127.16 ng·g−1, at 60 °C, and then decreased continuously. The content of the free HAs was the highest at 60 °C. For the protein-bound HAs, their content was found to generally decrease with the increase in smoking duration and temperature. However, at 5 h, the content of protein-bound HAs slightly increased to 984.2 ng·g−1. Meanwhile, at 90 °C, it increased to 1643.53 ng·g−1. Additionally, a total of 16 volatile organic compounds (VOCs) were found in all of the meat samples, of which 10 VOCs (one acid, three aldehydes and seven phenols) were significantly related to the formation of free HAs. These findings showed that all the different types of HAs were produced under low-temperature processing, which provided scientific insights into the potential generation of HAs during meat smoking processes and could be used as a reference to minimize the risks of cancer related to the consumption of smoked meat products.

The Shandong Peninsula is the largest peninsula in China and has played a vital part in Chinese civilisation. The ancient independent Laizi kingdom was located on the Shandong peninsula. While large demographic changes have happened at this peninsula throughout history, the genetic landscape of modern populations on this peninsula has never been clarified. The aims of our study were to investigate population genetic characteristics of the populations on the Shandong peninsula and to reveal their genetic affinities with other populations around the world. Allele frequencies, Hardy–Weinberg equilibrium, and forensic parameters of 15 autosomal STRs in the AmpFlSTR® Identifiler system were obtained from the studied populations with 2441 individuals in total. Allele frequencies were used to reveal the phylogenetic relationships among 287 worldwide populations. The combined power of discrimination (CPD) and the combined power of exclusion (CPE) in the whole of the Han population on the Shandong Peninsula were 0.999999999999999983 and 0.999998155, respectively. The 15 autosomal loci were polymorphic and informative among our studied populations. Genetic homogeneities were revealed between the modern populations on the Shandong Peninsula and Han nationalities from Northeastern China as well as East China.

Geminiviruses cause serious symptoms and devastating losses in crop plants. With a circular, single-stranded DNA genome, geminiviruses multiply their genomic DNA in the nucleus, requiring the nuclear shuttling of viral proteins and viral genomic DNAs. Many host factors, acting as proviral or antiviral factors, play key roles in geminivirus infections. Here, we report the roles of a tomato glutaredoxin (GRX), SlGRXC6, in the infection of Tomato yellow leaf curl virus (TYLCV), a single-component geminivirus. The V2 protein of TYLCV specifically and preferentially interacts with SlGRXC6 among the 55-member tomato GRX family that are broadly involved in oxidative stress responses, plant development, and pathogen responses. We show that overexpressed SlGRXC6 increases the nuclear accumulation of V2 by inhibiting its nuclear export and, in turn, inhibits trafficking of the V1 protein and viral genomic DNA. Conversely, the silenced expression of SlGRXC6 leads to an enhanced susceptibility to TYLCV. SlGRXC6 is also involved in symptom development as we observed a positive correlation where overexpression of SlGRXC6 promotes while knockdown of SlGRXC6 expression inhibits plant growth. We further showed that SlGRXC6 works with SlNTRC80, a tomato NADPH-dependent thioredoxin reductase, to regulate plant growth. V2 didn't interact with SlNTRC80 but competed with SlNTR80 for binding to SlGRXC6, suggesting that the V2-disrupted SlGRXC6-SlNTRC80 interaction is partially responsible for the virus-caused symptoms. These results suggest that SlGRXC6 functions as a host restriction factor that inhibits the nuclear trafficking of viral components and point out a new way to control TYLCV infection by targeting the V2-SlGRXC6 interaction.

A recessive gene PmQ conferring powdery mildew resistance was previously localized on the long arm of chromosome 2B in winter wheat landrace Qingxinmai. Breeder-friendly molecular markers are necessary for introgressing this gene into adapted wheat backgrounds for developing disease-resistant wheat cultivars. Three Kompetitive allele-specific PCR (KASP) markers were developed and validated based on SNP variants detected by Bulked segregant analysis-RNA-Seq (BSR-Seq) analysis using a recombinant inbred population derived from cross Qingxinmai × 041133. Two polymorphic SSR markers were also developed from the motifs in the Chinese Spring reference genome sequences of the target genomic region. Those markers were incorporated into a more saturated genetic linkage map for PmQ. The two flanking markers, Xicsq405 and Xicsk18, are 1.1 and 0.9 cM from PmQ, respectively. The KASP marker Xicsk19 produced unique amplification pattern in 158 out of 160 wheat cultivars or breeding lines. This marker with the gene-linked SSR markers Xicsqc, Xicsqd and Xicsq405 provides an efficient means in molecular marker-assisted selection for PmQ in wheat breeding. The corresponding genomic region of PmQ in the Chinese Spring reference genome has a conserved synteny with the genomes of sequenced wheat cultivars and Triticum durum, T. diccocoides, T. uratu, and barley. The annotation of the two genes, TraesCS2B01G517400.1 and TraesCS2B01G517700.1, associated with plant defense against pathogens placed a start for cloning PmQ.

Silicon nitride (Si3N4) is a promising biomaterial, currently used in spinal fusion implants. Such implants should result in high vertebral union rates without major complications. However, pseudarthrosis remains an important complication that could lead to a need for implant replacement. Making silicon nitride implants more bioactive could lead to higher fusion rates, and reduce the incidence of pseudarthrosis. In this study, it was hypothesized that creating a highly negatively charged Si3N4 surface would enhance its bioactivity without affecting the antibacterial nature of the material. To this end, samples were thermally, chemically, and thermochemically treated. Apatite formation was examined for a 21-day immersion period as an in-vitro estimate of bioactivity. Staphylococcus aureus bacteria were inoculated on the surface of the samples, and their viability was investigated. It was found that the thermochemically and chemically treated samples exhibited enhanced bioactivity, as demonstrated by the increased spontaneous formation of apatite on their surface. All modified samples showed a reduction in the bacterial population; however, no statistically significant differences were noticed between groups. This study successfully demonstrated a simple method to improve the in vitro bioactivity of Si3N4 implants while maintaining the bacteriostatic properties.

In order to provide more evidence for the evaluation of the ecological risks of transgenic maize, arthropod population dynamics and biodiversity in fields planted with two kinds of transgenic maize (DBN9868, expressing the PAT and EPSPS genes, and DBN9936, expressing the Cry1Ab and EPSPS gene) were investigated by direct observation and trapping for three years. The recorded arthropod species belonged to 19 orders and 87 families, including Aphidoidea, Chrysomelidae, Coccinellidae, Chrysopidae and Araneae. The species richness, Shannon–Wiener diversity index, Pielou evenness index, dominance index and community similarity index of arthropod communities in maize fields were statistically analyzed, and the results showed that (1) the biodiversity difference of arthropod communities between transgenic maize and non-transgenic maize was smaller than that between different conventional cultivars; (2) the differences between ground-dwelling arthropod communities were less obvious than those between plant-inhabiting arthropod communities; and (3) Lepidoptera, the target pests of Bt maize, were not the dominant population in maize fields, and the dominant arthropod population in maize fields varied greatly between years and months. Combining those results, we concluded that the transgenic maize DBN9868 and DBN9936 had no significant effect on the arthropod communities in the field.