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Abstract Abundant infiltration of tumor-associated macrophages (TAMs) within the tumor stroma plays a pivotal role in inducing immune escape in pancreatic cancer (PC). Lactate serves as a direct regulator of macrophage polarization and functions, although the precise regulation mechanism remains inadequately understood. Our study revealed that PC cells (PCs) promote macrophage polarization toward M2d through high lactate secretion. M2d is characterized by elevated secretion of IL-10 and VEGF-A, which diminish CD8+T cells cytotoxicity and promote tumor neoangiogenesis simultaneously. Additionally, we identify 2,5’-oligoadenylate synthase 3 (OAS3) as an essential regulator of M2d polarization, upregulated by PCs via lactate/METTL3/OAS3 axis. Increased OAS3 expression in TAMs correlates with m6A modification mediated by METTL3 on OAS3 mRNA and is associated with poorer prognosis in PC patients. OAS3 deficiency in macrophages substantially impairs IL-10highVEGF-AhighM2d polarization and their pro-tumor functions while enhancing the therapeutic efficacy of gemcitabine and anti-PD-L1 mAb in humanized mouse models. In conclusion, OAS3 presents as a promising immune therapeutic target for reversing IL-10highVEGF-AhighM2d infiltration and restoring CD8+T cell-mediated anti-tumor immunity in pancreatic cancer. Graphical abstract

Bladder cancer (BLCA) remains a significant health risk despite advancements in medical science that have led to reduced incidence and death rates. While the molecular regulatory mechanisms of BLCA are not yet fully understood, HSPE1, a member of the heat shock protein family, is regarded as a reliable prognostic target for BLCA. Using data from The Cancer Genome Atlas (TCGA) database, the differential expression levels of HSPE1 and its relationship to GPX4 were examined. Gene Set Enrichment Analysis was used to carry out HSPE1 pathway enrichment analysis. HSPE1 and GPX4 expressions in cells were assessed using quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and western blotting. Cell cycle alternations and apoptosis were evaluated using flow cytometry. Cell proliferation was assessed using EdU and colony formation assays. The lactate dehydrogenase (LDH) test, glutathione (GSH) measurement, and Liperfluo assay were utilized to evaluate the presence of ferroptosis in cells. BLCA tissues and cells had significantly elevated levels of HSPE1. In BLCA, high expression of HSPE1 inhibited apoptosis while promoting cell proliferation and cell cycle progression. Significant enrichment of HSPE1 was found in the GSH metabolism and ferroptosis pathways, according to pathway enrichment analysis. In cancer cells, HSPE1 promoted GSH accumulation, decreased lipid peroxidation, and inhibited cell ferroptosis, as demonstrated in a rescue experiment with the ferroptosis inhibitor Fer-1. Pearson correlation analysis unveiled a substantial positive correlation between HSPE1 and the ferroptosis regulator GPX4. According to the results of rescue experiments, HSPE1 regulated GPX4 to affect cell lipid peroxidation levels and GSH accumulation. HSPE1 plays a crucial role in regulating GPX4 to prevent BLCA cells from undergoing ferroptosis, with this control mechanism dependent on GSH.

IntroductionWe recently showed that sub-kilohertz electrical stimulation of the afferent somata in the dorsal root ganglia (DRG) reversibly blocks afferent transmission. Here, we further investigated whether similar conduction block can be achieved by stimulating the nerve trunk with electrical peripheral nerve stimulation (ePNS).MethodsWe explored the mechanisms and parameters of conduction block by ePNS via ex vivo single-fiber recordings from two somatic (sciatic and saphenous) and one autonomic (vagal) nerves harvested from mice. Action potentials were evoked on one end of the nerve and recorded on the other end from teased nerve filaments, i.e., single-fiber recordings. ePNS was delivered in the middle of the nerve trunk using a glass suction electrode at frequencies of 5, 10, 50, 100, 500, and 1000 Hz.ResultsSuprathreshold ePNS reversibly blocks axonal neural transmission of both thinly myelinated Aδ-fiber axons and unmyelinated C-fiber axons. ePNS leads to a progressive decrease in conduction velocity (CV) until transmission blockage, suggesting activity-dependent conduction slowing. The blocking efficiency is dependent on the axonal conduction velocity, with Aδ-fibers efficiently blocked by 50–1000 Hz stimulation and C-fibers blocked by 10–50 Hz. The corresponding NEURON simulation of action potential transmission indicates that the disrupted transmembrane sodium and potassium concentration gradients underly the transmission block by the ePNS.DiscussionThe current study provides direct evidence of reversible Aδ- and C-fiber transmission blockage by low-frequency (

Abstract Objective (1) This study aims to identify distinct serum metabolites in gastric cancer patients compared to healthy individuals, providing valuable insights into postoperative efficacy evaluation and monitoring of gastric cancer recurrence; (2) Methods: Serum samples were collected from 15 healthy individuals, 16 gastric cancer patients before surgery, 3 months after surgery, 6 months after surgery, and 15 gastric cancer recurrence patients. T-test and analysis of variance (ANOVA) were performed to screen 489 differential metabolites between the preoperative group and the healthy control group. Based on the level of the above metabolites in the recurrence, preoperative, three-month postoperative, and six-month postoperative groups, we further selected 18 significant differential metabolites by ANOVA and partial least squares discriminant analysis (PLS-DA). The result of hierarchical clustering analysis about the above metabolites showed that the samples were regrouped into the tumor-bearing group (comprising the original recurrence and preoperative groups) and the tumor-free group (comprising the original three-month postoperative and six-month postoperative groups). Based on the results of PLS-DA, 7 differential metabolites (VIP > 1.0) were further selected to distinguish the tumor-bearing group and the tumor-free group. Finally, the results of hierarchical clustering analysis showed that these 7 metabolites could well identify gastric cancer recurrence; (3) Results: Lysophosphatidic acids, triglycerides, lysine, and sphingosine-1-phosphate were significantly elevated in the three-month postoperative, six-month postoperative, and healthy control groups, compared to the preoperative and recurrence groups. Conversely, phosphatidylcholine, oxidized ceramide, and phosphatidylglycerol were significantly reduced in the three-month postoperative, six-month postoperative, and healthy control groups compared to the preoperative and recurrence groups. However, these substances did not show significant differences between the preoperative and recurrence groups, nor between the three-month postoperative, six-month postoperative, and healthy control groups; (4) Conclusions: Our findings demonstrate the presence of distinct metabolites in the serum of gastric cancer patients compared to healthy individuals. Lysophosphatidic acid, triglycerides, lysine, sphingosine-1-phosphate, phosphatidylcholine, oxidized ceramide, and phosphatidylglycerol hold potential as biomarkers for evaluating postoperative efficacy and monitoring recurrence in gastric cancer patients. These metabolites exhibit varying concentrations across different sample categories.

Panax japonicus C. A. Meyer is renowned for its significant therapeutic effects and is commonly used worldwide. Its active ingredients, triterpenoid saponins, show variation in content among different tissues. The tissue-specific distribution of saponins is potentially related to the expression of vital genes in the biosynthesis pathway. In this study, the contents of five saponins (ginsenoside Ro, chikusetsusaponin IV, chikusetsusaponin IVa, ginsenoside Rg1, and ginsenoside Rb1) in three different tissues were determined by HPLC. Transcriptome sequencing analysis identified differentially expressed genes (DEGs) involved in triterpenoid saponin biosynthesis, highlighting significant correlations between saponin contents and the expression levels of 10 cytochrome p450 monooxygenase (CYP) and 3 UDP-glycosyltransferase (UGT) genes. Cloning, sequencing, and prokaryotic expression of UGT genes confirmed the molecular weights of UGT proteins. Gene sequence alignment and phylogenetic analysis provided preliminary insights into UGT gene functions. Meanwhile, the function of one UGT gene was characterized in the yeast. These findings advance our understanding of the triterpenoid saponin biosynthesis in P. japonicus and support future research in traditional Chinese medicine (TCM) and synthetic biology.

Using Panax japonicus as research material, callus induction and culture were carried out, and high-yielding cell lines were screened to establish a suspension culture system that promotes callus growth and the accumulation of the “total saponins” (total content of triterpenoid glycosides or ginsenosides). Using the root as an explant, the medium for callus induction and proliferation was optimized by adjusting culture conditions (initial inoculation amount, carbon source, shaking speed, hormone concentration, culture time) and a high-yielding cell line with efficient proliferation and high total saponins content was screened out. The conditions of suspension culture were refined to find out the most suitable conditions for the suspension culture of callus, and finally, the suspension culture system was established. We found that the lowest (5%) contamination rate was achieved by disinfecting the fresh roots with 75% alcohol for 60 s, followed by soaking in 10% NaClO for 15 min. The highest induction rate (88.17%) of callus was obtained using the medium MS + 16.11 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 sucrose + 7.5 g·L−1 agar. The callus was loose when the callus subcultured on the proliferation medium (MS + 5.37 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 sucrose + 3.8 g·L−1 gellan gum) for 21 days. The callus growth was cultured in a liquid growth medium (MS + 5.37 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 sucrose) with an initial inoculation amount of 40 g·L−1, a shaking speed of 110 r/min and darkness. Cell growth was fastest with a culture period of 21 days. We replaced the growth medium with the production medium (MS + 5.37 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 glucose) for maximum accumulation of total saponins. [Conclusion] A callus induction and suspension culture system for the root of P. japonicus was established. In this way, we can promote the accumulation of total saponins in callus cells and provide a basis for large-scale cell culture and industrial production of medicinal total saponins.

This paper proposes a theoretical framework for assessing ecological protection and restoration from the perspective of ecological efficiency. We applied the super-efficiency Slack-based measure model to examine the social and economic impacts of ecological resource consumption transformation in Heilongjiang Province, China. Additionally, a convergence analysis was used to evaluate and test the impact of the standard deviation ellipse method on regional sustainability. The results indicated that the land use structure was unstable; the conversion rate of resource consumption was low; and the average Ecological efficiency was only 0.343 in terms of the land use structure. Funds for forest ecological restoration have a significant impact on the effectiveness of ecological resource transformation. Implementing the Chinese ecological restoration project improves the ecological efficiency level of the communities. The center of gravity of ecological efficiency moved greatly in the years when forestry investment increased. Technological transfer and diffusion, experience imitation in environmental regulation, and eventually convergent steady-state levels of the ecological efficiency of different regions are necessary to improve the economic and social development level of regions with low environmental quality efforts should be made to reduce resource consumption intensity, increase fund utilization efficiency, and form a comprehensive and systematic system of ecological environment governance through reasonable enhancement of regional environmental regulations, increased investment in technological advancement, and funds for ecological protection and restoration.

In recent years, offshore wind farms have frequently encountered engineering geological disasters such as seabed liquefaction and scouring. Consequently, in situ monitoring has become essential for the safe siting, construction, and operation of these installations. Current technologies are hampered by limitations in single-parameter monitoring and insufficient probe-penetration depth, hindering comprehensive multi-parameter dynamic monitoring of seabed sediments. To address these challenges, we propose a foldable multi-sensor probe and establish an underwater adaptive continuous penetration system capable of concurrently measuring seabed elevation changes and sediment pore water pressure profiles. The reliability of the equipment design is confirmed through static analysis of the frame structure and sealed cabin. Furthermore, laboratory tests validate the stability and accuracy of the electrical and mechanical sensor measurements. Preliminary tests conducted in a harbor environment demonstrate the system’s effectiveness.

This paper presents a two-stage method combining data envelopment analysis (DEA) and a Tobit model to analyze the comprehensive operating efficiency of 28 airports in China in 2016. At the first stage, the DEA-BCC (Banker-Charnes-Cooper) model was employed to obtain the comprehensive operating efficiency of the combination of flight departure punctuality, non-cancellations, landing bridge rates from the perspective of airport infrastructure, surrounding airspace, route layouts, flight volume and weather. At the second stage, a Tobit model was used to analyze the influence of nine input variables from four aspects on obtained comprehensive operating efficiency, ultimately providing a clear and straightforward basis for formulating and testing policies. The comprehensive operating efficiency with this combination was further compared with each of the three efficiencies respectively. The important findings included the following: (1) The comprehensive operation efficiencies of most airports were greater than the individual efficiency; (2) These four types of operation efficiencies for most airports did not achieved DEA validity (100% efficiency), except for six airports (i.e., Haikou, Dalian, Jinan, Fuzhou, Nanning and Lanzhou); (3) These factors affecting each of the four types of operation efficiencies were different in that the number of terminals, duration of impact and average daily inbound and outbound flights had a negative impact on airport operational efficiency, while the average number of overnight aircraft per day and peak hour sorties had positive effects.

Abstract Background Rectal cancer is one of the most lethal of gastrointestinal malignancies. Metabonomics has gradually developed as a convenient, inexpensive and non-destructive technique for the study of cancers. Methods A total of 150 tissue samples from 25 rectal cancer patients were analyzed by liquid chromatography–mass spectrometry (LC–MS), and 6 tissue samples were collected from each patient (group 1: tumor; group 2: 0.5 cm from tumor; group 3:1 cm from tumor; group 4:2 cm from tumor; group 5:3 cm from tumor and group 6:5 cm from tumor). The differential metabolites of tumor tissues and 5 cm from the tumor (normal tissues) were first selected. The differential metabolites between tumor tissues and normal tissues were regrouped by hierarchical clustering analysis, and further selected by discriminant analysis according to the regrouping of clustering results. The potential safe margin of clinical T(cT)1,cT2 stage rectal cancer and cT3,cT4 stage rectal cancer at the metabolomic level was further identified by observing the changes in the level of differential metabolites within the samples from group 1 to group 6. Results We found 22 specific metabolites to distinguish tumor tissue and normal tissue. The most significant changes in metabolite levels were observed at 0.5 cm (cT1, cT2) and 2.0 cm (cT3, cT4) from the tumor, while the changes in the tissues afterwards showed a stable trend. Conclusions There are differential metabolites between tumor tissues and normal tissues in rectal cancer. Based on our limited sample size, the safe distal incision margin for rectal cancer surgery in metabolites may be 0.5 cm in patients with cT1 and cT2 stage rectal cancer and 2.0 cm in patients with cT3 and cT4 stage rectal cancer.

Airports, as integral components of the global aviation industry, experience dynamic changes in air passenger traffic load, which are also related to the trends of local urban development and airspace restrictions of nearby airports. Using time-series data from 2006 to 2019, this study comprehensively applied the autoregressive distributed lag and vector auto-regression mode approaches to identify causal relationships between the urban development factors, including GDP, population, tourism industry, industrial structure, etc., of Tianjin city (China); the Tianjin airport passenger flow; and the Beijing Capital airport's airspace restriction factor, namely, airport aircraft sorties. The results show that the growth of Tianjin city's GDP, primary industry and disposable income per capita was accompanied by a long-term decline in the passenger flow at the Tianjin airport. In addition, increased aircraft sorties of Tianjin airport, as well as the growth of primary, secondary and tertiary industries in Tianjin city, led to a short-term decline in passenger flow at the Tianjin airport. In general, there is variability in the long- and short-term impacts of urban economic structure on airport passenger flow, and this variability applies to other airports. The increased aircraft sorties at the Beijing Capital airport had a short-term positive impact on passenger flow at the Tianjin airport but resulted in a long-term decline of the latter's aircraft sorties. This phenomenon indicates that there is interaction between airports and that this influence varies depending on the competition and cooperation mechanisms between airports. The findings of this study are considered instrumental in guiding the competitive and cooperative strategies of nearby airports and predicting the coupled trends of the airport and urban development.

IntroductionVisceral motor responses (VMR) to graded colorectal distension (CRD) have been extensively implemented to assess the level of visceral pain in awake rodents, which are inevitably confounded by movement artifacts and cannot be conveniently implemented to assess invasive neuromodulation protocols for treating visceral pain. In this report, we present an optimized protocol with prolonged urethane infusion that enables robust and repeatable recordings of VMR to CRD in mice under deep anesthesia, providing a two-hour window to objectively assess the efficacy of visceral pain management strategies.MethodsDuring all surgical procedures, C57BL/6 mice of both sexes (8–12 weeks, 25–35 g) were anesthetized with 2% isoflurane inhalation. An abdominal incision was made to allow Teflon-coated stainless steel wire electrodes to be sutured to the oblique abdominal musculature. A thin polyethylene catheter (Φ 0.2 mm) was placed intraperitoneally and externalized from the abdominal incision for delivering the prolonged urethane infusion. A cylindric plastic-film balloon (Φ 8 mm x 15 mm when distended) was inserted intra-anally, and its depth into the colorectum was precisely controlled by measuring the distance between the end of the balloon and the anus. Subsequently, the mouse was switched from isoflurane anesthesia to the new urethane anesthesia protocol, which consisted of a bout of infusion (0.6 g urethane per kg weight, g/kg) administered intraperitoneally via the catheter and continuous low-dose infusion throughout the experiment at 0.15–0.23 g per kg weight per hour (g/kg/h).ResultsUsing this new anesthesia protocol, we systematically investigated the significant impact of balloon depth into the colorectum on evoked VMR, which showed a progressive reduction with increased balloon insertion depth from the rectal region into the distal colonic region. Intracolonic TNBS treatment induced enhanced VMR to CRD of the colonic region (>10 mm from the anus) only in male mice, whereas colonic VMR was not significantly altered by TNBS in female mice.DiscussionConducting VMR to CRD in anesthetized mice using the current protocol will enable future objective assessments of various invasive neuromodulatory strategies for alleviating visceral pain.

A new magnetic adsorbent, namely, MnFe2O4/D201, with deep-treatment ability and high selectivity adsorption for As(V) was prepared. According to isotherm adsorption and kinetics, As(V) adsorption is primarily used for chemical bonding throughout the single-layer adsorbing process. The maximum As(V) adsorption capacity of MnFe2O4/D201 can reach 35.8 mg/g at pH 3. MnFe2O4/D201 also exhibits higher selectivity adsorption against Cl-, NO3-, SO42-, and PO43-. According to the thermodynamic results, the adsorption process was spontaneous and endothermic. The adsorption capacity is maintained at 81% of the initial after ten adsorption-desorption cycles. As(V) concentrations ranging from 1 mg/L to 10 μg/L can be treated in fixed-bed column experiments. The effectual cure volume of As(V) reaches 1332 BV (26.64 L). The removal mechanism primarily comprises electrostatic attraction and complexation.

Abstract Background and Aims Accumulating studies identified that BUB1 mitotic checkpoint serine/threonine kinase B (BUB1B) is integrally involved in the initiation and development of tumors. Nevertheless, the precise biological role and underlying mechanisms of BUB1B in hepatocellular carcinoma (HCC) remain indistinct. Method To figure out the role of BUB1B in HCC, we first assessed its expression using The Cancer Genome Atlas (TCGA) and Gene Expression Profiling Interactive Analysis (GEPIA) databases. We then verified BUB1B expression in HCC tissues, nontumor tissues, and HCC cell lines through western blotting, quantitative reverse transcription‐polymerase chain reaction, and immunohistochemistry. To explore the specific function of BUB1B in HCC in vivo and in vitro, we performed the flow cytometry, Cell Counting Kit‐8, 5‐ethynyl‐2′‐deoxyuridine incorporation, colony formation, Transwell, wound‐healing, subcutaneous tumor growth, and metastasis assays. Additionally, we identified the BUB1B‐regulated pathways involved in HCC by using gene set enrichment analysis. Results Our data displayed that higher BUB1B expression was detected in HCC tissues and HCC cell lines. The overexpression of BUB1B was positively correlated with adverse clinicopathological characteristics. Survival analyses showed that lower recurrence‐free and overall survival rates were correlated with the overexpression of BUB1B in patients with HCC. Moreover, the malignancy of HCC was facilitated by BUB1B both in vivo and in vitro. Lastly, the results were confirmed by western blots, which showed that BUB1B upregulated mTORC1 signaling pathway in HCC. Meanwhile, the oncogenic effect of BUB1B will be impaired when the mTORC1 signaling pathway was inhibited by rapamycin. Conclusion We highlighted that BUB1B played an oncogenic role in HCC and was identified as a possible clinical prognostic factor and a potential novel therapeutic target for HCC.

Improvisation is an effective way to cope with rapid changes and obtain unexpected opportunities in a complex environment. Based on the cognitive-affective system theory, this study investigates the dual mediating role of cognitive flexibility and emotional intelligence between shared leadership and improvisation and the moderating role of promotion focus. We used multilevel and multi-sourced data to test the theoretical model and used a social network approach to measure shared leadership in teams. Our sample was comprised of 40 teams and 240 team members. The empirical findings indicated that cognitive flexibility and emotional intelligence mediated the relationship between shared leadership and improvisation; promotion focus moderated the relationship between shared leadership and improvisation, and the mediation effect via cognitive flexibility. This study contributes to expanding on improvisation research from the perspective of shared leadership and incorporating both the cognitive and the emotional process of the generation of improvisation into a theoretical framework from a compound perspective, which will open the black box for the mediation mechanism from shared leadership to improvisation. Furthermore, promotion focus is introduced into the research and creatively corresponds to the cognition-affection mediation mechanism, which expands the applicable scope of the regulatory focus theory.

A rational urban spatial development pattern is necessary for China to optimize urban resource allocation and promote low-carbon urban development. Although research on carbon emission reduction has increased, few studies examine the impact of urban spatial development patterns from a spatial–structural perspective. This study uses static and dynamic spatial Durbin models to analyze the dynamic impact of urban spatial development patterns on carbon emissions based on municipal-level statistical data and LandScan high-resolution global population distribution data from 2004 to 2019. The empirical results show that (1) urban spatial development patterns characterized by coefficients of variation have an inverted U-shaped relationship with carbon emissions; (2) direct spatial spillover effects have a long-term U-shaped relationship, while indirect effects have an inverted U-shaped relationship; (3) by analyzing the heterogeneity of city sizes and geographical area, the smaller the city is, the more effectively the compact urban spatial development pattern reduces carbon emissions; and (4) compact urban spatial development patterns in the west suppress carbon emissions compared to the east and central regions. The findings of this paper have policy implications for optimizing spatial development patterns and achieving low-carbon development in Chinese cities.

The precise cellular patterning and decisions of early embryogenesis have been hard to mimic in vitro. Here, the authors culture murine embryonic and trophoblast stem cells together with extra-embryonic endoderm stem cells to form embryo-like structures (ETX-embryoids), which can initiate an implantation response.

Abstract It is not fully clear why there is a higher contribution of pluripotent stem cells (PSCs) to the chimera produced by injection of PSCs into 4-cell or 8-cell stage embryos compared with blastocyst injection. Here, we show that not only embryonic stem cells (ESCs) but also induced pluripotent stem cells (iPSCs) can generate F0 nearly 100% donor cell-derived mice by 4-cell stage embryo injection, and the approach has a “dose effect”. Through an analysis of the PSC-secreted proteins, Activin A was found to impede epiblast (EPI) lineage development while promoting trophectoderm (TE) differentiation, resulting in replacement of the EPI lineage of host embryos with PSCs. Interestingly, the injection of ESCs into blastocysts cultured with Activin A (cultured from 4-cell stage to early blastocyst at E3.5) could increase the contribution of ESCs to the chimera. The results indicated that PSCs secrete protein Activin A to improve their EPI competency after injection into recipient embryos through influencing the development of mouse early embryos. This result is useful for optimizing the chimera production system and for a deep understanding of PSCs effects on early embryo development.

Abstract Our study examined the in vivo chimeric and survival capacities of chimeras created by injecting tetraploid embryonic stem cells (ESCs) expressing green fluorescent protein (GFP) into diploid embryos. At 3.5 days post-coitum (dpc) and 4.5 dpc, the tetraploid ESCs were able to contribute to the inner cell mass (ICM) just as diploid ESCs tagged with GFP. At 6.5 dpc, 8.0 dpc and 10.5 dpc, the tetraploid ESCs manifested in the same location as the diploid ESCs. The GFP cells in the extraembryonic tissues and fetuses of tetraploid ESC chimeras were tetraploid as determined by fluorescence activated cell sorting (FACS). Furthermore, tetraploid ESCs contributed to the development of the placenta, embryolemma and umbilical cord at 13.5 dpc and 16.5 dpc; however, very less GFP cells were found in the fetuses of tetraploid ESC chimeras. We further found that the proliferation of tetraploid ESCs was slower than that of diploid ESCs. In addition, the relative mRNA expression in the three germ layers and the trophoblast was abnormal in the EBs of tetraploid ESCs compared with diploid ESCs. In short, slower proliferation and abnormal differentiation potential of tetraploid ESCs might be two of the reasons for their poor survival and chimeric capacities.

We experimentally isolated and characterized a CFEM protein with putative GPI-anchored site BcCFEM1 in Botrytis cinerea. BcCFEM1 contains a CFEM (common in several fungal extracellular membrane proteins) domain with the characteristic eight cysteine residues at N terminus, and a predicted GPI modification site at C terminus. BcCFEM1 was significantly up-regulated during early stage of infection on bean leaves and induced chlorosis in Nicotiana benthamiana leaves using Agrobacterium infiltration method. Targeted deletion of BcCFEM1 in B. cinerea affected virulence, conidial production and stress tolerance, but not growth rate, conidial germination, colony morphology, and sclerotial formation. However, over expression of BcCFEM1 did not make any observable phenotype change. Therefore, our data suggested that BcCFEM1 contributes to virulence, conidial production, and stress tolerance. These findings further enhance our understanding on the sophisticated pathogenicity of B. cinerea beyond necrotrophic stage, highlighting the importance of CFEM protein to B. cinerea and other broad-host-range necrotrophic pathogens.

Mining at the fully mechanized working face below the goaf of the short-distance coal seam is influenced by the upper goaf. To address this problem, methods such as theoretical analyses, numerical simulation, and on-site measurement are used to study the strata behavior characteristics of the Ningxia Lingxin Coal Mine 051508 working face in this study. The roof weighting intervals of the working faces below the goaf and the non-goaf are obtained via theoretical calculations. The stoping processes of the working faces below the goaf and the non-goaf are simulated with FLAC3D to obtain the distribution law of the bearing pressure and plastic zones before the working face. Based on the statistical analysis of the measured working resistance of the supports and its distribution, the roof weighting interval of the working face mining below the goaf is obtained. The results show that the roof weighting interval and the advanced abutment pressure during mining at the working face below the goaf are smaller than those below the non-goaf, providing a reasonable theoretical basis for mining below the goaf, and having important significance for safe and efficient mining.

Soybean cyst nematode (SCN), Heterodera glycines, is the most devastating pathogen of soybean worldwide. MicroRNAs (miRNAs) are a class of small, non-coding RNAs that are known to play important role in plant stress response. However, there are few reports profiling the miRNA expression patterns during pathogen stress. We sequenced four small RNA libraries from two soybean cultivar (Hairbin xiaoheidou, SCN race 3 resistant, Liaodou 10, SCN race 3 susceptible) that grown under un-inoculated and SCN-inoculated soil. Small RNAs were mapped to soybean genome sequence, 364 known soybean miRNA genes were identified in total. In addition, 21 potential miRNA candidates were identified. Comparative analysis of miRNA profiling indicated 101 miRNAs belong to 40 families were SCN-responsive. We also found 20 miRNAs with different express pattern even between two cultivars of the same species. These findings suggest that miRNA paly important role in soybean response to SCN and have important implications for further identification of miRNAs under pathogen stress.

The enteric nervous system (ENS) functions largely independently of the central nervous system (CNS). Glutamate, the dominant neurotransmitter in the CNS and sensory afferents, is not a primary neurotransmitter in the ENS. Only a fraction (-2%) of myenteric neurons in the mouse distal colon and rectum (colorectum) are positive for vesicular glutamate transporter type 2 (VGLUT2), the structure and function of which remain undetermined. Here, we systematically characterized VGLUT2-positive enteric neurons (VGLUT2-ENs) through sparse labeling with adeno-associated virus, single-cell mRNA sequencing (scRNAseq), and GCaMP6f calcium imaging. Our results reveal that the majority of VGLUT2-ENs (29 of 31, 93.5%) exhibited Dogiel type I morphology with a single aborally projecting axon; most axons (26 of 29, 89.7%) are between 4 and 10 mm long, each traversing 19 to 34 myenteric ganglia. These anatomical features exclude the VGLUT2-ENs from being intrinsic primary afferent or motor neurons. The scRNA-seq conducted on 52 VGLUT2-ENs suggests different expression profiles from conventional descending interneurons. Ex vivo GCaMP6f recordings from flattened colorectum indicate that almost all VGLUT2-EN (181 of 215, 84.2%) are indirectly activated by colorectal stretch via nicotinic cholinergic neural transmission. In conclusion, VGLUT2-ENs are a functionally unique group of enteric neurons with single aborally projecting long axons that traverse multiple myenteric ganglia and are activated indirectly by colorectal mechanical stretch. This knowledge will provide a solid foundation for subsequent studies on the potential interactions of VGLUT2-EN with extrinsic colorectal afferents via glutamatergic neurotransmission. [ABSTRACT FROM AUTHOR]

Improving corporate green innovation is the key support for achieving sustainable economic development under the concept of green development. However, existing research lacks a theoretical framework and empirical evidence on how public procurement affects corporate green innovation. Drawing on institutional and stakeholder theory, this study examines whether and how public procurement affects corporate green innovation. Using a sample of Chinese listed companies from 2010 to 2020, we find a positive correlation between public procurement and firm's green innovation. Further analysis reveals that corporate social responsibility (CSR) acts as a mediator between public procurement and business green innovation. Additionally, we argue that CEO green experience and environmental regulation strengthen the positive relationship between public procurement and green innovation. This study contributes to the theoretical understanding of how public procurement influences corporate green innovation and offers valuable insights for public procurement agencies and enterprises in practice. [ABSTRACT FROM AUTHOR]