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Abstract Background Saffron (Crocus sativus L.) is a valuable herb. With the increasing demand for saffron, people are starting to focus on how to increase its yields. Intercropping and microbial interactions have a positive effect on plant yield, including enhanced soil fertility, enriched microbial diversity, reduced pest and disease incidences, and improved plant growth. However, the impact of intercropping saffron with other plants on saffron yields and soil microbial community diversity remains unclear. In our study, we counted the number of saffron flowers in two cropping patterns (saffron monoculture and saffron-grape intercropping), and analyzed the microbial community diversity and composition using Illumina high-throughput sequencing methods based on 16 S and ITS amplicons. Results The results showed that saffron-grape intercropping significantly increased number of flowers compared to saffron monoculture (P

Potassium (K+) plays a role in enzyme activation, membrane transport, and osmotic regulation processes. An increase in potassium content can significantly improve the elasticity and combustibility of tobacco and reduce the content of harmful substances. Here, we report that the expression analysis of Nt GF14e, a 14-3-3 gene, increased markedly after low-potassium treatment (LK). Then, chlorophyll content, POD activity and potassium content, were significantly increased in overexpression of Nt GF14e transgenic tobacco lines compared with those in the wild type plants. The net K+ efflux rates were severely lower in the transgenic plants than in the wild type under LK stress. Furthermore, transcriptome analysis identified 5708 upregulated genes and 2787 downregulated genes between Nt GF14e overexpressing transgenic tobacco plants. The expression levels of some potassium-related genes were increased, such as CBL-interacting protein kinase 2 (CIPK2), Nt CIPK23, Nt CIPK25, H+-ATPase isoform 2 a (AHA2a), Nt AHA4a, Stelar K+ outward rectifier 1(SKOR1), and high affinity K+ transporter 5 (HAK5). The result of yeast two-hybrid and luciferase complementation imaging experiments suggested Nt GF14e could interact with CIPK2. Overall, these findings indicate that NtGF14e plays a vital roles in improving tobacco LK tolerance and enhancing potassium nutrition signaling pathways in tobacco plants.

Abstract Land-use change is the main driver of carbon storage change in terrestrial ecosystems. Currently, domestic and international studies mainly focus on the impact of carbon storage changes on climate, while studies on the impact of land-use changes on carbon storage in complex terrestrial ecosystems are few. The Jialing River Basin (JRB), with a total area of ~ 160,000 km2, diverse topography, and elevation differences exceeding 5 km, is an ideal case for understanding the complex interactions between land-use change and carbon storage dynamics. Taking the JRB as our study area, we analyzed land-use changes from 2000 to 2020. Subsequently, we simulated land-use patterns for business-as-usual (BAU), cropland protection (CP), and ecological priority (EP) scenarios in 2035 using the PLUS model. Additionally, we assessed carbon storage using the InVEST model. This approach helps us to accurately understand the carbon change processes in regional complex terrestrial ecosystems and to formulate scientifically informed land-use policies. The results revealed the following: (1) Cropland was the most dominant land-use type (LUT) in the region, and it was the only LUT experiencing net reduction, with 92.22% of newly designated construction land originating from cropland. (2) In the JRB, total carbon storage steadily decreased after 2005, with significant spatial heterogeneity. This pattern was marked by higher carbon storage levels in the north and lower levels in the south, with a distinct demarcation line. The conversion of cropland to construction land is the main factor driving the reduction in carbon storage. (3) Compared with the BAU and EP scenarios, the CP scenario demonstrated a smaller reduction in cropland area, a smaller addition to construction land area, and a lower depletion in the JRB total carbon storage from 2020 to 2035. This study demonstrates the effectiveness of the PLUS and InVEST models in analyzing complex ecosystems and offers data support for quantitatively assessing regional ecosystem services. Strict adherence to the cropland replenishment task mandated by the Chinese government is crucial to increase cropland areas in the JRB and consequently enhance the carbon sequestration capacity of its ecosystem. Such efforts are vital for ensuring the food and ecological security of the JRB, particularly in the pursuit of the “dual-carbon” objective.

Abstract Purpose Rot disease caused by Fusarium poses a formidable threat to the growth of saffron (Crocus sativus L.), resulting in substantial damage to both yield and quality. It is paramount to delve into the root causes of rot disease in saffron to optimize both yield and quality. Existing preventive and treatment modalities have exerted deleterious effects on corms and the natural environment. Consequently, the quest for efficacious and eco-friendly methods such as biological control agents has become an urgent imperative. Methods The disparate distribution of microbial communities between rhizospheric microorganisms and saffron serves as the foundational exploration for uncovering the underlying causes of rot disease. Samples from various saffron organs and rhizosphere soil were gathered, and the sequencing data from the microbial communities were interpreted using 16S rRNA and ITS gene sequencing methods. This facilitated an in-depth examination of the composition and changes of microorganisms in both healthy and diseased saffron plants. Results The findings indicated rot disease reduced the abundance and diversity of microorganisms in saffron, and the fungal co-occurrence networks were less stable and their communities were more sensitive to rot disease than the bacterial community. Fusarium was the predominant genus in diseased samples, accounting for 99.19% and 89.77% of the communities in diseased leaves and corms. With corms and leaves displaying heightened susceptibility to infection compared to other plant organs. Some of the beneficial bacterial taxa enriched in the diseased plants were also identified in networks, they showed an antagonistic relationship with Fusarium, suggesting a potential for these bacteria to be used in biologically based control strategies against rot disease. These insights could prove invaluable for the development of biocontrol agents aimed at combating this plant ailment. Conclusion These findings significantly advance our understanding of saffron-microbiome interactions and could provide fundamental and important data for improving saffron yield and quality in the process of sustainable development.

Abstract Background Emerging evidences suggest that aberrant metabolites contributes to the immunosuppressive microenvironment that leads to cancer immune evasion. Among tumor immunosuppressive cells, myeloid-derived suppressor cells (MDSCs) are pathologically activated and extremely immunosuppressive, which are closely associated with poor clinical outcomes of cancer patients. However, the correlation between MDSCs mediated immunosuppression and particular cancer metabolism remained elusive. Methods Spontaneous lung adenocarcinoma and subcutaneous mouse tumor models, gas chromatography–mass spectrometry (GC–MS) and immunofluorescence assay of patient-derived lung adenocarcinoma tissues, and flow cytometry, RNA sequencing and Western blotting of immune cells, were utilized. Results Metabolite profiling revealed a significant accumulation of acetic acids in tumor tissues from both patients and mouse model, which contribute to immune suppression and cancer progression significantly through free fatty acid receptor 2 (FFAR2). Furthermore, FFAR2 is highly expressed in the myeloid-derived suppressor cells (MDSCs) from the tumor of lung adenocarcinoma (LUAD) patients which is greatly associated with poor prognosis. Surprisingly, whole or myeloid Ffar2 gene deletion markedly inhibited urethane-induced lung carcinogenesis and syngeneic tumor growth with reduced MDSCs and increased CD8+ T cell infiltration. Mechanistically, FFAR2 deficiency in MDSCs significantly reduced the expression of Arg1 through Gαq/Calcium/PPAR-γ axis, which eliminated T cell dysfunction through relieving L-Arginine consumption in tumor microenvironment. Therefore, replenishment of L-Arginine or inhibition to PPAR-γ restored acetic acids/FFAR2 mediated suppression to T cells significantly. Finally, FFAR2 inhibition overcame resistance to immune checkpoint blockade through enhancing the recruitment and cytotoxicity of tumor-infiltrating T cells. Conclusion Altogether, our results demonstrate that the acetic acids/FFAR2 axis enhances MDSCs mediated immunosuppression through Gαq/calcium/PPAR-γ/Arg1 signaling pathway, thus contributing to cancer progression. Therefore, FFAR2 may serve as a potential new target to eliminate pathologically activated MDSCs and reverse immunosuppressive tumor microenvironment, which has great potential in improving clinical outcomes of cancer immunotherapy.

Background N-Ethylmaleimide (NEM), an agonist of the potassium chloride cotransporters 2 (KCC2) receptor, has been correlated with neurosuppressive outcomes, including decreased pain perception and the prevention of epileptic seizures. Nevertheless, its relationship with sleep-inducing effects remains unreported. Objective The present study aimed to investigate the potential enhancement of NEM on the sleep-inducing properties of alprazolam (Alp). Methods The test of the righting reflex was used to identify the appropriate concentrations of Alp and NEM for inducing sleep-promoting effects in mice. Total sleep duration and sleep quality were evaluated through EEG/EMG analysis. The neural mechanism underlying the sleep-promoting effect was examined through c-fos immunoreactivity in the brain using immunofluorescence. Furthermore, potential CNS-side effects of the combination Alp and NEM were assessed using LABORAS automated home-cage behavioral phenotyping. Results Combination administration of Alp (1.84 mg/kg) and NEM (1.0 mg/kg) significantly decreased sleep latency and increased sleep duration in comparison to administering 1.84 mg/kg Alp alone. This effect was characterized by a notable increase in REM duration. The findings from c-fos immunoreactivity indicated that NEM significantly suppressed neuron activation in brain regions associated with wakefulness. Additionally, combination administration of Alp and NEM showed no effects on mouse neural behaviors during automated home cage monitoring. Conclusions This study is the first to propose and demonstrate a combination therapy involving Alp and NEM that not only enhances the hypnotic effect but also mitigates potential CNS side effects, suggesting its potential application in treating insomnia.

Among the bioactive compounds, lipid-soluble tanshinone is present in Salvia miltiorrhiza, a medicinal plant species. While it is known that ethephon has the ability to inhibit the tanshinones biosynthesis in the S. miltiorrhiza hairy root, however the underlying regulatory mechanism remains obscure. In this study, using the transcriptome dataset of the S. miltiorrhiza hairy root induced by ethephon, an ethylene-responsive transcriptional factor EIN3-like 1 (SmEIL1) was identified. The SmEIL1 protein was found to be localized in the nuclei, and confirmed by the transient transformation observed in tobacco leaves. The overexpression of SmEIL1 was able to inhibit the tanshinones accumulation to a large degree, as well as down-regulate tanshinones biosynthetic genes including SmGGPPS1, SmHMGR1, SmHMGS1, SmCPS1, SmKSL1 and SmCYP76AH1. These are well recognized participants in the tanshinones biosynthesis pathway. Further investigation on the SmEIL1 was observed to inhibit the transcription of the CPS1 gene by the Dual-Luciferase (Dual-LUC) and yeast one-hybrid (Y1H) assays. The data in this work will be of value regarding the involvement of EILs in regulating the biosynthesis of tanshinones and lay the foundation for the metabolic engineering of bioactive ingredients in S. miltiorrhiza.

Potatoes are widely planted worldwide and are the third most important food crop. Mutation breeding involves the artificial use of various physical and chemical factors to induce plants to produce new genotypes. In this study, we established a potato radiation mutation system and analysed the features of a radiation mutant. The LD50 of the potato callus was 24.8 Gy after linear regression analysis. The radiation mutant 1 (RM1) showed significant dwarfism and strong growth; RM1 plants decreased in height by 31%. The root and leaf fresh weights of the RM1 increased 1.8-fold compared to the wild-type (WT, Chuanyu 10) cultivar. Leaf microstructure results showed that the thicknesses of the upper epidermis and lower epidermis of RM1 plants were greater than those of the WT cultivar. Transcriptome analysis of seeding revealed 1179 upregulated and 1641 downregulated differentially expressed genes (DEGs) in RM1 plants. Further analysis showed that the expression levels of WRKY and MYB transcription factors, CIPK and MAPK protein kinases, ABC transporters, hormones, and ROS pathway genes were altered. These results provide theoretical support for potato breeding research and enrich the functional network of vital breeding-related genes.

CBL-interacting protein kinases (CIPKs) play important regulatory roles in plant growth development and abiotic stress tolerance. However, the biological roles of these genes in response to low-nitrate (LN) stress in potato plants have not been determined. Here, we reported that StCIPK23 was expressed mainly in roots and leaves. StCIPK23 was located mainly in the cell membrane, nucleus, and cytoplasm. Further research suggested that, compared with wild-type (WT) plants, StCIPK23-overexpressing plants were taller and had significantly greater nitrate and ammonium nitrogen contents under LN stress. StCIPK23 overexpression can increase StAT, StNRT2.1, StNR, StGS1-3, and StGOGAT expression levels in StCIPK23 transgenic seedlings compared to those in WT plants under LN stress. The results of yeast two-hybrid and luciferase complementation imaging experiments suggested that StCIPK23 could interact with StCBL3. Real-time reverse transcription–PCR revealed the StCIPK23 expression level peaked at 6 h and the StCBL3 expression level peaked at 9 h in the roots under LN stress. In conclusion, we found that StCIPK23 and StCBL3 form a complex to regulate the expression of key genes in the nitrogen metabolism pathway to improve LN tolerance in potato plants.

The BTB/POZ (broad-complex, tramtrack, and bric-a-brac) family of proteins is widespread in plants and animals and plays important roles in growth, development, metabolism, and environmental responses. There are few reports on BTB family genes in potato. In this study, 34 sequences containing conserved BTB domains were obtained from the potato gene database, and the phylogenetic, physical, and chemical properties, gene structure, conserved motif, domain, and chromosomal localization of the potato BTB protein family were analyzed via bioinformatics methods. In addition, we used qRT-PCR to detect 12 selected StBTB genes. The results confirmed that these genes are involved in cold, ABA, salt, hydrogen peroxide (H2O2), drought, and low-nitrogen stress, which is highly important for elucidating BTB family members and studying stress response and tolerance mechanisms. This study provides a theoretical basis for the study of the function and expression of potato BTB and lays a solid foundation for further understanding the molecular mechanism of the potato BTB gene under various environmental stresses.

Plants and insects are the most diverse and ecologically important organisms in the terrestrial biosphere. Their interactions are also among the richest biotic relationships, and offer significant insights into the evolution of terrestrial ecosystem complexity through the geological record. This investigation of the late Rhaetian Xujiahe and the earliest Jurassic Zhenzhuchong floral assemblages provides the first data on foliar herbivory generated by terrestrial arthropods across the Triassic–Jurassic transition in the eastern Tethys (East Asia) region. The damage types from two fossil assemblages are collectively attributed to seven functional feeding and egg-laying categories (i.e., hole feeding, margin feeding, surface feeding, skeletonization, piercing and sucking, oviposition, and galling). Most feeding strategies are spread across the major plant groups and persist through the Triassic–Jurassic boundary, with the exception of skeletonization (a category of external foliage feeding), which was restricted to the latest Triassic within dipteridacean ferns. The survey reveals that the respective frequency and diversity of interactions between plants and insects prior to and following the end-Triassic mass extinction event are almost the same, despite a substantial turnover of floral components. This suggest that insect herbivores were largely able to transfer to alternative (but commonly related) plant groups during the dramatic floristic turnover and environmental changes at the end of the Triassic. Sporadic occurrences of foliar modifications, such as marginal cusps on pinnules of Pterophyllum and prominent ridges on the rachises of some ferns and bennettites are interpreted as adaptations for defense against insect herbivores. A few differences in taxonomic composition and herbivory representation between the latest Triassic Xujiahe flora and the earliest Jurassic Zhenzhuchong flora are more likely to be related to collection and preservational biases rather than reflecting palaeoecological changes. We encourage further investigations exploring the distribution of insect damage in fossil floras from other palaeolatitudinal zones and spanning other major extinction events to develop a better understanding of terrestrial ecosystem responses to major crises in Earth’s history.

BackgroundGitelman Syndrome (GS) patients frequently exhibit disrupted glucose metabolism, attributed to hypokalemia, hypomagnesemia and heightened aldosterone. This study delved into the genetic underpinnings linked to insulin resistance and diabetes in a GS patient, contextualized within his family history.MethodsThe hydrochlorothiazide and furosemide loading test were performed to ascertain the presence of GS. Oral glucose tolerance test (OGTT) evaluated glucose metabolism and insulin sensitivity. Whole-exome sequencing, validated by Sanger sequencing, was employed to confirm gene mutations, which were then tracked among the patient’s relatives.ResultsSymptoms and laboratory examination confirmed the clinical diagnosis of GS. Comprehensive whole-exome sequencing, augmented by Sanger sequencing validation, revealed a compound heterozygous mutation within the SLC12A3 gene (c.1108G>C in exon 9, c.676G>A in exon 5 and c.2398G>A in exon 20) in the patient. The OGTT affirmed diabetes and heightened insulin resistance, distinct from previous patients with GS we evaluated. Further genetic analysis identified a missense heterozygous mutation (c.97C>G in exon 1) within the PDX1 gene, inherited from the patient’s diabetic mother without GS. Furthermore, the patient’s brother, with impaired glucose tolerance but regular potassium levels, also bore this mutation, hinting at additional impacts of the PDX1 gene mutation on glucose metabolism regulation beyond the known impacts of GS.ConclusionThis study unveils unprecedented compound heterozygous mutations in the SLC12A3 and PDX1 genes in a GS patient. These findings illuminate the potential complex genetic factors influencing glucose metabolism disruptions in GS.Take-home messageThis research uncovers a novel combination of SLC12A3 and PDX1 gene mutations in a Gitelman Syndrome patient, revealing intricate genetic factors that potentially disrupt glucose metabolism and shedding light on familial diabetes links.

IntroductionAcute lung injury (ALI) is a severe respiratory tract disorder facilitated by dysregulated inflammation, oxidative stress and intestinal ecosystem. Fecal microbiota transplantation (FMT) is a rapid method for gut microbiota (GM) reconstruction. Furthermore, our previous studies have confirmed that human umbilical cord mesenchymal stromal cells (HUC-MSCs) can alleviate ALI by improving GM composition. Therefore, we aimed to explore the efficacy and mechanism of FMT from HUC-MSCs-treated mice on ALI.MethodsIn brief, fresh feces from HUC-MSCs-treated mice were collected for FMT, and the mice were randomly assigned into NC, FMT, LPS, ABX-LPS, and ABX-LPS-FMT groups (n = 12/group). Subsequently, the mice were administrated with antibiotic mixtures to deplete GM, and given lipopolysaccharide and FMT to induce ALI and rebuild GM. Next, the therapeutic effect was evaluated by bronchoalveolar lavage fluid (BALF) and histopathology. Immune cells in peripheral blood and apoptosis in lung tissues were measured. Furthermore, oxidative stress- and inflammation-related parameter levels were tested in BALF, serum, lung and ileal tissues. The expressions of apoptosis-associated, TLR4/NF-κB pathway-associated, Nrf2/HO-1 pathway related and tightly linked proteins in the lung and ileal tissues were assessed. Moreover, 16S rRNA was conducted to assess GM composition and distribution.ResultsOur results revealed that FMT obviously improved the pathological damage of lung and ileum, recovered the immune system of peripheral blood, decreased the cell apoptosis of lung, and inhibited inflammation and oxidative stress in BALF, serum, lung and ileum tissues. Moreover, FMT also elevated ZO-1, claudin-1, and occludin protein expressions, activating the Nrf2/HO-1 pathway but hindering the TLR4/NF-κB pathway. Of note, the relative abundances of Bacteroides, Christensenella, Coprococcus, and Roseburia were decreased, while the relative abundances of Xenorhabdus, Sutterella, and Acinetobacter were increased in the ABX-LPS-FMT group.ConclusionFMT from HUC-MSCs-treated mice may alleviate ALI by inhibiting inflammation and reconstructing GM, additionally, we also found that the TLR4/NF-κB and Nrf2/HO-1 pathways may involve in the improvement of FMT on ALI, which offers novel insights for the functions and mechanisms of FMT from HUC-MSCs-treated mice on ALI.

Although many carriers for the delivery of chemotherapeutic drugs have been investigated, the disadvantages of passive targeting and uncontrolled drug release limit their utility. Herein, hyaluronic acid (HA) was hydrophobically modified to serve as a carrier for binding to cluster determinant 44 (CD44) overexpressed on tumor cell surfaces. Specifically, after deacetylation, HA was grafted to dodecylamine or tetradecylamine to afford amphiphilic zwitterionic polymer micelles, designated dHAD and dHAT, respectively, for the delivery of paclitaxel (PTX). The micelles were negatively charged at pH 7.4 and positively charged at pH 5.6, and this pH sensitivity facilitated PTX release under acidic conditions. The cell uptake efficiencies of the dHAD-PTX and dHAT-PTX micelles by MCF-7 cells after 4 h of incubation were 96.9% and 95.4%, respectively, and their affinities for CD44 were twice that of HA. Furthermore, the micelles markedly inhibited tumor growth both in vitro and in vivo, with IC50 values of 1.943 μg/mL for dHAD-PTX and 1.874 μg/mL for dHAT-PTX for MCF-7 cells; the tumor inhibition rate of dHAD-PTX (92.96%) was higher than that of dHAT-PTX (78.65%). Importantly, dHAD and dHAT micelles showed negligible systemic toxicity. Our findings suggest that these micelles are promising delivery vehicles for antitumor drugs.

Research and promotion of green technology can improve energy efficiency and help to optimize energy structure, which are key considerations in achieving the “dual carbon” goals of carbon peaking and carbon neutrality. Owing to the “dual externality” arising from green technology innovation (GTI), government policies have an important impact on this area. Environmental taxes and government subsidies are important means for governments to improve GTI and optimize energy utilization, with different environmental taxes and subsidies having different impacts. This study evaluates the impact of environmental taxes and government subsidies in China from 2009 to 2019, and assesses how these policies interacted with each other to determine China’s efficiency in GTI over this period. The study’s findings based on analysis of 30 provinces and cities in China indicate that environmental taxes have a threshold effect on the development of green technologies, and that the current level of taxation is insufficient to encourage optimized development in this area. Government subsidies can also help to reduce the negative effects arising from the current environmental tax burden. To optimize government policy implementation in this area, improve the level of GTI, and improve energy use efficiency, this study proposes countermeasures based on differentiated policies, including gradually increasing the rate and scope of environmental taxation, and strengthening the supervision of government subsidies.

Abstract Background Ampullary adenocarcinoma (AAC) arises from the ampulla of Vater where the pancreatic duct and bile duct join and empty into the duodenum. It can be classified into intestinal and pancreatobiliary types based on histopathology or immunohistochemistry. However, there are no biomarkers for further classification of pancreatobiliary-type AAC which has important implications for its treatment. We aimed to identify the tumor origin of pancreatobiliary-type AAC by systematically analyzing whole-slide images (WSIs), survival data, and genome sequencing data collected from multiple centers. Methods This study involved three experiments. First, we extracted quantitative and highly interpretable features from the tumor region in WSIs and constructed a histologic classifier to differentiate between pancreatic adenocarcinoma (PAC) and cholangiocarcinoma. The histologic classifier was then applied to patients with pancreatobiliary-type AAC to infer the tumor origin. Secondly, we compared the overall survival of patients with pancreatobiliary-type AAC stratified by the adjuvant chemotherapy regimens designed for PAC or cholangiocarcinoma. Finally, we compared the mutation landscape of pancreatobiliary-type AAC with those of PAC and cholangiocarcinoma. Results The histologic classifier accurately classified PAC and cholangiocarcinoma in both the internal and external validation sets (AUC > 0.99). All pancreatobiliary-type AACs (n = 45) were classified as PAC. The patients with pancreatobiliary-type AAC receiving regimens designed for PAC showed more favorable overall survival than those receiving regimens designed for cholangiocarcinoma in a multivariable Cox regression (hazard ratio = 7.24, 95% confidence interval: 1.28–40.78, P = 0.025). The results of mutation analysis showed that the mutation landscape of AAC was very similar to that of PAC but distinct from that of cholangiocarcinoma. Conclusions This multi-center study provides compelling evidence that pancreatobiliary-type AAC resembles PAC instead of cholangiocarcinoma in different aspects, which can guide the treatment selection and clinical trials planning for pancreatobiliary-type AAC.

Tetrodotoxin (TTX) is an exceedingly toxic non-protein biotoxin that demonstrates remarkable selectivity and affinity for sodium channels on the excitation membrane of nerves. This property allows TTX to effectively obstruct nerve conduction, resulting in nerve paralysis and fatality. Although the mechanistic aspects of its toxicity are well understood, there is a dearth of literature addressing alterations in the neural microenvironment subsequent to TTX poisoning. In this research endeavor, we harnessed human pluripotent induced stem cells to generate cerebral organoids—an innovative model closely mirroring the structural and functional intricacies of the human brain. This model was employed to scrutinize the comprehensive transcriptomic shifts induced by TTX exposure, thereby delving into the neurotoxic properties of TTX and its potential underlying mechanisms. Our findings revealed 455 differentially expressed mRNAs (DEmRNAs), 212 differentially expressed lncRNAs (DElncRNAs), and 18 differentially expressed miRNAs (DEmiRNAs) in the TTX-exposed group when juxtaposed with the control cohort. Through meticulous Gene Ontology (GO) annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, and protein–protein interaction (PPI) analysis, we ascertained that these differential genes predominantly participate in the regulation of voltage-gated channels and synaptic homeostasis. A comprehensive ceRNA network analysis unveiled that DEmRNAs exert control over the expression of ion channels and neurocytokines, suggesting their potential role in mediating apoptosis.

With the aging of many populations, cognitive and motor dysfunction caused by ischemic stroke (IS) secondary to long-term chronic cerebral ischemia presents a global problem. Enriched environment (EE), a classic paradigm of environment response and genetic interaction, has shown tremendous influence on the brain. This research aimed to investigate the potential effect of EE on cognitive and motor function in mice with chronic cerebral ischemia and secondary IS. In the chronic cerebral hypoperfusion (CCH) phase, EE treatment improved behavior performance by alleviating neuronal loss and white matter myelin damage, promoting the expression of brain-derived neurotrophic factor (BDNF) and phosphor-cAMP response element binding protein (p-CREB). Furthermore, infiltration of microglia/macrophages and astrocytes was inhibited, and the levels of IL-1β and TNFα were decreased. In the IS phase, EE altered the neuronal outcome on day 21 but not on day one after IS. In addition, EE inhibited IS-induced infiltration of microglia/macrophages and astrocytes, mediated the polarization of microglia/macrophages, and reduced pro-inflammatory factors. Importantly, EE improved IS-induced cognitive and motor deficits on day 21. Collectively, our work demonstrates that EE protects mice from cognitive and motor dysfunction and inhibits neuroinflammation caused by CCH and IS.

The work shows that volcanic-related elevated continental chemical weathering could have played a significant role in global environmental perturbations during the Triassic-Jurassic mass extinction.

Ischemic stroke is one of the world’s leading causes of death and disability. It has been established that gender differences in stroke outcomes prevail, and the immune response after stroke is an important factor affecting patient outcomes. However, gender disparities lead to different immune metabolic tendencies closely related to immune regulation after stroke. The present review provides a comprehensive overview of the role and mechanism of immune regulation based on sex differences in ischemic stroke pathology.

Nitrogen is one of the most important elements for improving potato yield. However, excessive application of nitrogenous fertilizer not only produces river and other environmental pollution but also increases agricultural production costs. In recent years, to explain the molecular mechanisms of nitrogen metabolites, some vital genes involved have been reported; however, only limited success has been achieved in potato. Here, we report that the expression of cytosolic glyceraldehyde-3-phosphate dehydrogenase 1 (StGAPC1) is increased under low-nitrogen stress. StGAPC1-overexpressing potato seedlings had more biomass and a significant increase in total nitrogen content and root nitrate influx rate compared to the wild type. The overexpression of StGAPC1 also increased the expression of nitrate transporters and increased ROS system activity to reduce hydrogen peroxide content under low-nitrogen stress. Our results provide a foundation for further research on StGAPC1 function in nitrogen absorption and utilization mechanisms in potato.

Adenylate kinase (ADK) is a key enzyme that is widely distributed in animals and plants. It plays an important role in growth and stress response. However, ADK genes in potato (StADK) have been little reported. It is of great significance to identify ADK members and understand the molecular mechanism of stress response and tolerance. Based on the potato genome data, 23 StADK genes were identified at a genome-wide level. We then performed a comprehensive study using a bioinformatics method. The results of the evolutionary tree showed that StADK proteins were divided into four groups, and they were highly homologous to the Arabidopsis thaliana ADK members. Meanwhile, our study found that they existed on eight chromosomes, and we obtained three pairs of fragment duplications. Furthermore, we detected the six selected StADK genes using qRT-PCR, and the results confirmed that the genes are involved in the regulation of cold, ABA, salt, H2O2 and drought stresses. Our study provides a theoretical basis for studying the function of the potato ADK genes and lays a solid foundation for further understanding the molecular mechanism of the potato ADK genes under various environmental stresses.

Potato tubers are rich in starch, vitamins, protein, minerals, and other nutrients. However, tuber sprouting produces solanine and reduces the commodity value of potatoes during storage. At present, it is known that some plant essential oils can inhibit tuber sprouting. It has been reported that grape seed oil (GSO) has antioxidant, anti-inflammatory, and anticancer characteristics, reducing blood lipids and delaying aging. In this study, we found for the first time that GSO delayed tuber sprouting, and the soluble sugar content and peroxidase activity changed after 60 days of GSO treatment. Furthermore, a comparative proteomic analysis of tuber bud eyes showed that after 30 days of GSO treatment, there were 206 and 129 differentially abundant proteins (DAPs) with increased and decreased abundance levels, respectively. After analysis, we found that 15 ROS-related proteins and 14 proteins involved in energy metabolism were DAPs. Among them, gamma aminobutyrate transaminase 1 had decreased abundance after GSO treatment. Meanwhile, the transcription level of genes related to GABA synthesis increased significantly according to qRT-PCR analysis. Our results provide new approaches to the proteomic mechanism of potato sprouting after GSO treatment and provide a theoretical basis for the application of GSO in inhibiting potato seed sprouts.

Novel nereistoxin derivatives containing phosphonate were synthesized and characterized via 31P, 1H and 13C NMR and HRMS. The anticholinesterase activity of the synthesized compounds was evaluated on human acetylcholinesterase (AChE) using the in vitro Ellman method. Most of the compounds exhibited good inhibition of acetylcholinesterase. All of these compounds were selected to assess their insecticidal activity (in vivo) against Mythimna separata Walker, Myzus persicae Sulzer and Rhopalosiphum padi. Most of the tested compounds displayed potent insecticidal activity against these three species. Compound 7f displayed good activity against all three insect species, showing LC50 values of 136.86 μg/mL for M. separata, 138.37 μg/mL for M. persicae and 131.64 μg/mL for R. padi. Compound 7b had the highest activity against M. persicae and R. padi, with LC50 values of 42.93 μg/mL and 58.19 μg/mL, respectively. Docking studies were performed to speculate the possible binding sites of the compounds and explain the reasons for the activity of the compounds. The results showed that the compounds had lower binding energies with AChE than with the acetylcholine receptor (AchR), suggesting that compounds are more easily bound with AChE.

Spirea (Spiraea sp.) plants are popular landscape plants in Utah and the Intermountain West United States. Spiraea betulifolia, S. japonica, S. media, S. nipponica, and S. thunbergii were evaluated for salinity tolerance in a greenhouse experiment. Plants were irrigated weekly with a nutrient solution at an electrical conductivity (EC) of 1.2 dS·m−1 (control) or saline solution at an EC of 3.0 or 6.0 dS·m−1 for 8 weeks. At the end of the experiment, all spirea plants survived and retained good visual quality, with average visual scores greater than 4 (0 = dead, 5 = excellent) when irrigated with saline solution at an EC of 3.0 dS·m−1, with the exception of S. thunbergii, which showed slight foliar salt damage and an average visual score of 3.8. When irrigated with saline solution at an EC of 6.0 dS·m−1, all S. thunbergii plants died, S. media exhibited severe foliar salt damage and an average visual score of 1.5, and S. betulifolia, S. japonica, and S. nipponica displayed slight-to-moderate foliar salt damage and average visual scores greater than 3. Regardless of spirea species, shoot dry weight decreased by 20% and 48% when irrigated with saline solution at ECs of 3.0 and 6.0 dS·m−1, respectively, compared with the control. Saline solution at an EC of 3.0 dS·m−1 did not affect net photosynthesis (Pn) of all spirea species except S. nipponica, but saline solution at an EC of 6.0 dS·m−1 decreased the Pn of all species by 36% to 60%. There were 37, 7, 36, 21, and 104 times more sodium (Na+) concentrations in leaf and 29, 28, 28, 13, and 69 times more chloride (Cl−) concentrations in leaf than in the control when S. betulifolia, S. japonica, S. media, S. nipponica, and S. thunbergii were irrigated with saline solution at an EC of 6.0 dS·m−1. Correlation analyses indicated that foliar salt damage and reduced plant growth and photosynthesis were induced mainly by Cl− ions accumulated in the spirea leaves. S. thunbergii was the most sensitive species; it had high mortality and low visual quality at both salinity levels. Spiraea japonica, S. nipponica, and S. betulifolia were relatively more tolerant and had good visual quality at elevated salinity compared with S. media and S. thunbergii. These research results are valuable for growers and landscape professionals during plant selection for nursery production using low-quality water and landscapes in salt-prone areas.

Sarin is a potent organophosphorus nerve agent that causes cognitive dysfunction, but its underlying molecular mechanisms are poorly understood. In this study, a rat model of repeated low-level sarin exposure was established using the subcutaneous injection of 0.4 × LD50 for 21 consecutive days. Sarin-exposed rats showed persistent learning and memory impairment and reduced hippocampal dendritic spine density. A whole-transcriptome analysis was applied to study the mechanism of sarin-induced cognitive impairment, and a total of 1035 differentially expressed mRNA (DEmRNA), including 44 DEmiRNA, 305 DElncRNA, and 412 DEcircRNA, were found in the hippocampus of sarin-treated rats. According to Gene Ontology (GO) annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and Protein–Protein Interaction (PPI) analysis, these DERNAs were mainly involved in neuronal synaptic plasticity and were related to the pathogenesis of neurodegenerative diseases. The circRNA/lncRNA–miRNA–mRNA ceRNA network was constructed, in which Circ_Fmn1, miR-741-3p, miR-764-3p, miR-871-3p, KIF1A, PTPN11, SYN1, and MT-CO3 formed one circuit, and Circ_Cacna1c, miR-10b-5p, miR-18a-5p, CACNA1C, PRKCD, and RASGRP1 constituted another circuit. The balance between the two circuits was crucial for maintaining synaptic plasticity and may be the regulatory mechanism by which sarin causes cognitive impairment. Our study reveals the ceRNA regulation mechanism of sarin exposure for the first time and provides new insights into the molecular mechanisms of other organophosphorus toxicants.

Sucrose phosphate synthase (SPS), a crucial rate-limiting enzyme that catalyzes the synthesis of precursors of sucrose, plays an indispensable role in the regulation of seed potato vigor. Nonetheless, the genes that encode SPS in potato have not undergone complete analysis, and the primary genes responsible for the regulation of seed potato vigor have not been screened and identified. In this study, four StSPS family members were identified by comparing the potato genome database with homologous proteins. Using bioinformatics, the physicochemical properties, subcellular localization, sequence structure, conserved motifs, and phylogenetics of StSPS were analyzed. The expression levels of StSPS in different potato tissues, from dormancy to sprouting in the seed potato tubers, were measured via qRT-PCR. The expression data from the potato genome database and previous transcriptome and proteome studies of dormancy to sprouting were also compared. After combining the analysis of SPS enzyme activity in diverse tuber tissues and the correlation analysis between multiple varieties with different dormancy periods and the expression of StSPS1, the primary gene StSPS1 that might regulate seed potato vigor was identified. This study set a theoretical and experimental groundwork for further verification and clarification of the regulatory function of StSPS1 in alterations in seed potato vigor.

TUBA1C, a microtubule component, contributes to the development of several cancers. Our purpose was to study the expression of TUBA1C, its potential prognostic value, and its effects on the infiltration of immune cells of low-grade glioma (LGG). Through applying multiple bioinformatics analyses, we extracted and analyzed datasets from TCGA, TIMER, GTEx, GEPIA, and HPA to investigate the potential oncogenic mechanisms of TUBA1C, including the correlation between TUBA1C and prognosis, immune-checkpoints, tumor microenvironment (TME), and infiltration of immune cells in LGG. GO functional annotations and KEGG pathway analyses were further applied to investigate the potential action of TUBA1C in LGG. We revealed that the mRNA levels of TUBA1C were increased in LGG tumor tissues than in normal tissues. Additionally, TUBA1C was up-regulated in the grade III of LGG than in grade II. Moreover, we found that TUBA1C may be an independent prognostic factor of LGG, and high TUBA1C expression correlated to a poor prognosis of LGG. TUBA1C expression was positively associated with the infiltration of B cells, CD8 T+ cells, CD4+ T cells, macrophages, dendritic cells, and neutrophils. TUBA1C was also verified to be co-expressed with immune-related genes and immune-checkpoints. GO and KEGG pathway analyses indicated that TUBA1C may potentially regulate the pathogenesis of LGG through immune-related pathways, including chemokine pathway; JAK-STAT pathway; natural killer cell mediated cytotoxicity; T cell receptor pathway; leukocyte migration; negative regulation of immune system process; regulation of lymphocyte activation; T cell activation and other pathways. In conclusion, TUBA1C expression is increased in LGG and high TUAB1C expression is related to a poor prognosis. TUBA1C may influence tumor development by regulating the tumor-infiltrating cells in the TME. TUBA1C may be a potential target for immunotherapy.

Abstract Background Saffron crocus (Crocus sativus) is a valuable spice with medicinal uses in gynaecopathia and nervous system diseases. Identify flowering regulatory genes plays a vital role in increasing flower numbers, thereby resulting in high saffron yield. Results Two full length transcriptome gene sets of flowering and non-flowering saffron crocus were established separately using the single-molecule real-time (SMRT) sequencing method. A total of sixteen SMRT cells generated 22.85 GB data and 75,351 full-length saffron crocus unigenes on the PacBio RS II panel and further obtained 79,028 SSRs, 72,603 lncRNAs and 25,400 alternative splicing (AS) events. Using an Illumina RNA-seq platform, an additional fifteen corms with different flower numbers were sequenced. Many differential expression unigenes (DEGs) were screened separately between flowering and matched non-flowering top buds with cold treatment (1677), flowering top buds of 20 g corms and non-flowering top buds of 6 g corms (1086), and flowering and matched non-flowering lateral buds (267). A total of 62 putative flower-related genes that played important roles in vernalization (VRNs), gibberellins (G3OX, G2OX), photoperiod (PHYB, TEM1, PIF4), autonomous (FCA) and age (SPLs) pathways were identified and a schematic representation of the flowering gene regulatory network in saffron crocus was reported for the first time. After validation by real-time qPCR in 30 samples, two novel genes, PB.20221.2 (p = 0.004, r = 0.52) and PB.38952.1 (p = 0.023, r = 0.41), showed significantly higher expression levels in flowering plants. Tissue distribution showed specifically high expression in flower organs and time course expression analysis suggested that the transcripts increasingly accumulated during the flower development period. Conclusions Full-length transcriptomes of flowering and non-flowering saffron crocus were obtained using a combined NGS short-read and SMRT long-read sequencing approach. This report is the first to describe the flowering gene regulatory network of saffron crocus and establishes a reference full-length transcriptome for future studies on saffron crocus and other Iridaceae plants.

Spirea (Spiraea sp.) plants are commonly used in landscapes in Utah and the intermountain western United States. The relative salt tolerance of seven japanese spirea (Spiraea japonica) cultivars (Galen, Minspi, NCSX1, NCSX2, SMNSJMFP, Tracy, and Yan) were evaluated in a greenhouse. Plants were irrigated with a nutrient solution with an electrical conductivity (EC) of 1.2 dS·m−1 (control) or saline solutions with an EC of 3.0 or 6.0 dS·m−1 once per week for 8 weeks. At 8 weeks after the initiation of treatment, all japanese spirea cultivars irrigated with saline solution with an EC of 3.0 dS·m−1 still exhibited good or excellent visual quality, with all plants having visual scores of 4 or 5 (0 = dead, 1 = severe foliar salt damage, 2 = moderate foliar salt damage, 3 = slight foliar salt damage, 4 = minimal foliar salt damage, 5 = excellent), except for Tracy and Yan, with only 29% and 64%, respectively, of plants with visual scores less than 3. When irrigated with saline solution with an EC of 6.0 dS·m−1, both ‘Tracy’ and ‘Yan’ plants died, and 75% of ‘NCSX2’ plants died. ‘Minspi’ showed severe foliar salt damage, with 32% of plants having a visual score of 1; 25% of plants died. ‘Galen’ and ‘NCSX1’ had slight-to-moderate foliar salt damage, with 25% and 21%, respectively, of plants with visual scores of 2 or less. However, 64% of ‘SMNSJMFP’ plants had good or excellent visual quality, with visual scores more than 4. Saline irrigation water with an EC of 3.0 dS·m−1 decreased the shoot dry weight of ‘Galen’, ‘Minspi’, ‘SMNSJMFP’, and ‘Yan’ by 27%, 22%, 28%, and 35%, respectively, compared with that of the control. All japanese spirea cultivars had 35% to 56% lower shoot dry weight than the control when they were irrigated with saline irrigation water with an EC of 6.0 dS·m−1. The japanese spirea were moderately sensitive to the salinity levels in this experiment. ‘Galen’ and ‘SMNSJMFP’ japanese spirea exhibited less foliar salt damage and reductions in shoot dry weight and were relatively more tolerant to the increased salinity levels tested in this study than the remaining five cultivars (Minspi, NCSX1, NCSX2, Tracy, and Yan).

BackgroundSaffron crocus (Crocus sativus) is an expensive and valuable species that presents preventive and curative effects. This study aimed to screen the key proteins affecting the floral initiation of saffron under cold stress and thus increasing yield by regulating the temperature.ResultsProtein expression profiles in flowering and non-flowering saffron buds were established using isobaric tags for relative or absolute quantitation (iTRAQ). A total of 5,624 proteins were identified, and 201 differentially abundant protein species (DAPs) were further obtained between the flowering and non-flowering groups. The most important functions of the upregulated DAPs were “sucrose metabolic process,” “lipid transport,” “glutathione metabolic process,” and “gene silencing by RNA.” Downregulated DAPs were significantly enriched in “starch biosynthetic process” and several oxidative stress response pathways. Three new flower-related proteins, CsFLK, CseIF4a, and CsHUA1, were identified in this study. The following eight key genes were validated by real-time qPCR in flowering and non-flowering top buds from five different growth phases: floral induction- and floral organ development-related genes CsFLK, CseIF4A, CsHUA1, and CsGSTU7; sucrose synthase activity-related genes CsSUS1 and CsSUS2; and starch synthase activity-related genes CsGBSS1 and CsPU1. These findings demonstrate the important roles played by sucrose/starch biosynthesis pathways in floral development at the mRNA level. During normal floral organ development, the sucrose contents in the top buds of saffron increased, and the starch contents decreased. In contrast, non-flowering buds showed significantly decreased sucrose contents under cold stress and no significant changes in starch contents compared with those in the dormancy stage.ConclusionIn this report, the protein profiles of saffron under cold stress and a normal environment were revealed for the first time by iTRAQ. A possible “reactive oxygen species–antioxidant system–starch/sugar interconversion flowering pathway” was established to explain the phenomenon that saffron does not bloom due to low temperature treatment.

This paper presents an experimental investigation on the effect of the replacement of natural coarse aggregate (NCA) with either recycled concrete aggregate (RCA) or recycled clay brick aggregate (RBA) on the compressive strengths of the hardened concrete. Two grades (C25 and C50) of concrete were investigated, which were achieved by using different water-to-cement ratios. In each grade concrete five different replacement rates, 0%, 25%, 50%, 75% and 100% were considered. In order to improve the performance of the recycled aggregates in the concrete mixes, the RCA and RBA were carefully sieved by using the optimal degradation. In this way the largest reduction in the 28-day compressive strength was found to be only 7.2% and 9.6% for C25 and C50 recycled concrete when the NCA was replaced 100% by RCA, and 11% and 13% for C25 and C50 recycled concrete when the NCA was replaced 100% by RBA. In general, the concrete with RCA has better performance than the concrete with RBA. The comparison of the present experimental results with those reported in literature for hardened concrete with either RCA or RBA demonstrates the effectiveness in improving the compressive strength by using the optimal gradation of recycled aggregates. Keywords: Recycled aggregate concrete, Waste concrete aggregate, Clay brick aggregate, Mechanical properties, Experiments

Abstract Non-small cell lung cancer (NSCLC) is one of the most common malignancies worldwide. In this study, we investigated Ultrahigh Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry and Gas Chromatography Time-of-Flight/Mass Spectrometry-based non-targeted metabolomic profiles of serum samples obtained from early-stage NSCLC patients and healthy controls (HC). Metabolic pathways and the biological relevance of potential biomarkers were extensively studied to gain insights into dysregulated metabolism in NSCLC. The identified biomarker candidates were further externally validated via a targeted metabolomics analysis. The global metabolomics profiles could clearly distinguish NSCLC patients from HC. Phosphatidylcholine (PC) levels were found to be dysregulated in glycerophospholipid (GPL) metabolism, which was the top altered pathway in early-stage NSCLC. Compared with those in HC, significant increases in the levels of saturated and monounsaturated PCs such as PC (15:0/18:1), PC (18:0/16:0) and PC (18:0/20:1) were observed in NSCLC. Additionally, relative to those in HC, the levels of 9 polyunsaturated PCs, namely, PC (17:2/2:0), PC (18:4/3:0), and PC (15:0/18:2), and so on were significantly decreased in NSCLC patients. A panel of 12 altered PCs had good diagnostic performance in differentiating early-stage NSCLC patients from HC, and these PCs may thus be used as serum biomarkers for the early diagnosis of NSCLC.

Chinese sacbrood virus (CSBV) infects Apis cerana larvae, resulting in the inability of the larvae to pupate and their consequent death, which may pose a serious threat to entire colonies. As there is no effective medical treatment for CSBV infections, further studies are necessary. In this study, an effective treatment for CSBV is described, based on a specific immunoglobulin Y (IgY) from egg yolk against CSBV. The inactivated vaccine was produced by ultracentrifugation and formalin treatment, using CSBV purified from a natural outbreak. The specific IgY was produced by immunization of white leghorn hens with the vaccine. An enzyme-linked immunosorbent assay using purified CSBV as the coating antigen revealed that the anti-CSBV IgY titer began increasing in the egg yolk on the 14th day post-immunization, reaching a peak on day 42, and anti-CSBV IgY remained at a high level until day 91. IgY isolated from the combinations of egg yolk collected between days 42–91 was purified by PEG and ammonium sulfate precipitation. In three repeated protection experiments using A. cerana larvae inoculated with CSBV, the survival rate of larvae was more than 80%, and the titer of anti-CSBV IgY was more than 25 and 24 when the larvae were fed IgY 24 h after and before inoculation with CSBV, respectively. Therefore, 400 colonies infected with CSBV were treated by feeding sugar containing IgY solutions with an antibody titer of 25, and the cure rate was 95–100%. Three hundred susceptible colonies were protected by feeding the larvae with sugar containing IgY solutions with an antibody titer of 24, and the protection rate was 97%. The results clearly suggest that a specific IgY was obtained from hens immunized with an inactivated-CSBV vaccine; this may be a novel method for controlling CSBV infection.

Being an important probability distribution with moderate skewness, the two-parameter gamma distribution is widely used in statistics. However, the maximum likelihood estimators (MLEs) of its parameters do not have closed forms, making them difficult to be implemented in applications. Moreover, the MLE of its shape parameter has low estimation efficiency due to its considerable bias. Thus, many other estimators have been investigated in the literature. We propose an easy computation of the MLEs in this article, where the MLE of the shape parameter is modified to be highly efficient and significantly better than most of existing estimators. [ABSTRACT FROM AUTHOR]