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ABSTRACT Background Lung adenocarcinoma (LUAD) is a fatal disease with metabolic abnormalities. The dysregulation of S100 calcium‐binding protein A2 (S100A2), a member of the S100 protein family, is connected to the development of various cancers. The impact of S100A2 on the LUAD occurrence and metastasis, however, has not yet been reported. The functional mechanism of S100A2 on LUAD cell metastasis was examined in this article. Methods The expression of TFAP2A and S100A2 in LUAD tissues and cells was analyzed by bioinformatics and qRT‐PCR, respectively. The enrichment pathway analysis was performed on S100A2. Bioinformatics analysis determined the binding relationship between TFAP2A and S100A2, and their interaction was validated through dual‐luciferase and chromatin immunoprecipitation experiments. Cell viability was determined using cell counting kit‐8 (CCK‐8). A transwell assay was performed to analyze the invasion and migration of cells. Immunofluorescence was conducted to obtain vimentin and E‐cadherin expression, and a western blot was used to detect the expression of MMP‐2, MMP‐9, GLS, and GLUD1. The kits measured the NADPH/NADP ratio, glutathione (GSH)/glutathione disulfide (GSSG) levels, and the contents of glutamine, α‐KG, and glutamate. Results S100A2 was upregulated in LUAD tissues and cells, and S100A2 mediated glutamine metabolism to induce LUAD metastasis. Additionally, the transcriptional regulator TFAP2A was discovered upstream of S100A2, and TFAP2A expression was upregulated in LUAD, which indicated that TFAP2A promoted the S100A2 expression. The rescue experiment found that upregulation of S100A2 could reverse the inhibitory effects of silencing TFAP2A on glutamine metabolism and cell metastasis. Conclusion In conclusion, by regulating glutamine metabolism, the TFAP2A/S100A2 axis facilitated LUAD metastasis. This suggested that targeting S100A2 could be beneficial for LUAD treatment.

Background & Aims: Acetaminophen (APAP) overdose is the most common cause of drug-induced liver injury worldwide. Uric acid (UA) is involved in sterile inflammation in many organs, but its role in APAP-induced liver injury remains elusive. Methods: We quantified the concentration of UA in the serum and liver tissues of APAP-overdosed mice and explored the changes in proteins involved in UA synthesis, absorption, and degeneration on APAP stimulation. We also examined the effects of inhibiting hepatocyte UA synthesis or reabsorption on APAP-induced liver injury in mice. Furthermore, we explored the process of UA clearance by peripheral macrophages. Results: APAP overdose significantly increased intrahepatic UA contents, which occurred earlier than apparent hepatocyte injury in APAP-overdosed mice. APAP overdose induced significant DNA leakage and may thereby increase the substrate of UA synthesis. APAP overdose also significantly increased the enzymatic activity of xanthine oxidase and urate oxidase and decreased the expression of the UA reabsorption transporter GLUT9 in hepatocytes. Inhibiting hepatocyte UA synthesis by febuxostat or reabsorption by hepatic-specific knockout of GLUT9 alleviated APAP-induced liver injury. Further experiments showed that monosodium urate but not soluble UA may be a major form of UA mediating hepatocyte injury. Additionally, monosodium urate further recruited circulating macrophages into the liver and then aggravated inflammation by increasing the levels of inflammatory factors and reactive oxygen species. Deletion of macrophages significantly ameliorated APAP-induced liver injury in mice. Conclusions: APAP overdose induces excessive UA production and leads to local high concentrations in the liver, which further injures cells and induces liver inflammation. Inhibiting the production of UA may be a potential therapeutic option for treating APAP-induced liver injury.

Abstract Background Since China adopted a policy to eliminate rural learning centers, boarding has become an important feature of the current rural student community. However, there is a lack of consensus on the impact of boarding schools on students' cognitive and non-cognitive development. This study investigates the effect of boarding schools on the development of cognitive and non-cognitive abilities of junior high school students in rural northwest China. Methods Using a sample of 5,660 seventh-grade students from 160 rural junior high schools across 19 counties, we identify a causal relationship between boarding and student abilities with the instrumental variables (IV) approach. Results The results suggest that boarding positively influences memory and attention, while it has no significant effect on other cognitive abilities such as reasoning, transcription speed, and accuracy. Furthermore, we find no significant association between boarding and the development of non-cognitive skills. Conclusions Given the widespread prevalence of boarding schools in rural regions, our study highlights the growing importance of improving school management to promote the development of students’ cognitive abilities and integrating the development of non-cognitive or social-emotional abilities into students’ daily routines.

Concerns persist about attentional engagement in online learning. The inter-subject correlation of eye movements (ISC) has shown promise as an accessible and effective method for attention assessment in online learning. This study extends previous studies investigating ISC of eye movements in online learning by addressing two research questions. Firstly, can ISC predict students’ attentional states at a finer level beyond a simple dichotomy of attention states (e.g., attending and distracted states)? Secondly, whether learners’ learning styles affect ISC’s prediction rate of attention assessment in video learning? Previous studies have shown that learners of different learning styles have different eye movement patterns when viewing static materials. However, limited research has explored the impact of learning styles on viewing patterns in video learning. An eye tracking experiment with participants watching lecture videos demonstrated a connection between ISC and self-reported attention states at a finer level. We also demonstrated that learning styles did not significantly affect ISC’s prediction rate of attention assessment in video learning, suggesting that ISC of eye movements can be effectively used without considering learners’ learning styles. These findings contribute to the ongoing discourse on optimizing attention assessment in the evolving landscape of online education.

An improved understanding of the environmental transmission of Taenia spp. is key to control of the parasite. Methods to detect and quantify Taenia eggs in different environmental matrices, including sludge and water, currently lack performance validation with regard to the recovery efficiency and process ease of use. Therefore, this study aimed to assess the recovery efficiency and process duration of commonly used methods for the detection of Taenia eggs in sludge and water samples. Ten detection methods for Taenia spp. eggs were selected from a systematic review. Sludge and water samples were spiked with a high dose of Taenia saginata eggs, i.e., around 200 eggs/g sludge and 50 eggs/ml water, and were tested using five methods each. The two methods with the highest egg recovery efficiencies were selected per matrix for assessment with a lower spiking dose, i.e., 4 eggs/g sludge and 1 egg/ml water. Each time five replicates were used. Recovery efficiency was defined as the proportion of the number of eggs recovered to the total number of eggs spiked. Using the high spiking dose, all samples tested positive for all the methods. The mean egg recovery efficiency varied from 4% to 69% for sludge samples and from 3% to 68% for water samples. Using the lower spiking dose, one of the methods performed on sludge samples was able to detect all replicates, whereas only one replicate was positive using the other method. For water, all low dose samples tested positive using both methods. In conclusion, most methods performed inadequately in recovering Taenia eggs from sludge and water, with half of the methods performed on the high dose samples having a mean egg recovery efficiency of approximately 10% or less. The assessed recovery methods were generally time-consuming and labourious. A more thorough validation of existing recovery methods and improvement of method protocols to increase recovery efficiency is thus urgently needed.

Awake rodent fMRI is becoming a promising non-invasive brain imaging module when investigating large-scale brain function given behavioral tasks. Previous studies have either applied sedatives during scanning or pre-treatment of anesthetics, e.g., isoflurane, to reduce the motion of animals, which could confound the brain function of “awake” states in rodents. Here, we have established a long training awake mouse fMRI-pupillometry paradigm/setup without the initial use of anesthesia. To validate the awake mouse fMRI platform, evoked BOLD-fMRI was performed to identify brain activation in the visual cortex, dorsal lateral geniculate nuclei, and superior colliculus. Furthermore, pupil signal fluctuation was investigated during scanning, showing a less dilated pupil after 5–8 weeks of intermittent training. Thus, using the awake mouse fMRI with real-time pupillometry provides a longitudinal functional mapping tool to study fully conscious mice.

Abstract Background Sarcocystis species are obligatorily heteroxenous parasites, of which some are zoonotic, representing a public health and economic impact. This study investigated the occurrence of Sarcocystis spp. in cattle sampled from a Belgian slaughterhouse. Methods A total of 200 carcasses were included in the study, sampled during 10 sampling days. The sedimentation method was applied to isolate the sarcocysts from both heart and diaphragm muscles collected from each carcass. Multiplex PCR, PCR–RFLP as well as cox1 gene sequencing techniques were applied serially on collected sarcocysts for species identification. Results Sarcocystis spp. were detected in 64% (128/200; 95% CI 57–71%) of the sampled carcasses. Female dairy cattle presented the highest Sarcocystis occurrence rate (91%) as well as the highest Sarcocystis species diversity compared to female beef and male beef. Sarcocystis spp. were detected more often in the heart muscles than in the diaphragm among female beef (p

Despite extensive efforts to increase the signal-to-noise ratio (SNR) of fMRI images for brain-wide mapping, technical advances of focal brain signal enhancement are lacking, in particular, for animal brain imaging. Emerging studies have combined fMRI with fiber optic-based optogenetics to decipher circuit-specific neuromodulation from meso to macroscales. High-resolution fMRI is needed to integrate hemodynamic responses into cross-scale functional dynamics, but the SNR remains a limiting factor given the complex implantation setup of animal brains. Here, we developed a multimodal fMRI imaging platform with an implanted inductive coil detector. This detector boosts the tSNR of MRI images, showing a 2–3-fold sensitivity gain over conventional coil configuration. In contrast to the cryoprobe or array coils with limited spaces for implanted brain interface, this setup offers a unique advantage to study brain circuit connectivity with optogenetic stimulation and can be further extended to other multimodal fMRI mapping schemes.

Abstract Avian pathogenic Escherichia coli (APEC), a pathotype of extraintestinal pathogenic E. coli, causes one of the most serious infectious diseases of poultry and shares some common virulence genes with neonatal meningitis-associated E. coli. TonB-dependent receptors (TBDRs) are ubiquitous outer membrane β-barrel proteins; they play an important role in the recognition of siderophores during iron uptake. Here, in the APEC strain DE205B, we investigated the role of four putative TBDRs—ireA, 0007, 0008, and 2235—in iron uptake. Glutathione-S-transferase pulldown assays indicated that the proteins encoded by these genes directly interact with TonB. Moreover, the expression levels of all four genes were significantly upregulated under iron-depleted conditions compared with iron-rich conditions. The expression levels of several iron uptake-related genes were significantly increased in the ireA, 0007, 0008, and 2235 deletion strains, with the upregulation being the most prominent in the ireA deletion mutant. Furthermore, iron uptake by the ireA deletion strain was significantly increased compared to that by the wild-type strain. Moreover, a tonB mutant strain was constructed to study the effect of tonB deletion on the TBDRs. We found that regardless of the presence of tonB, the expression levels of the genes encoding the four TBDRs were regulated by fur. In conclusion, our findings indicated that ireA, 0007, 0008, and 2235 indeed encode TBDRs, with ireA having the most important role in iron uptake. These results should help future studies explore the mechanisms underlying the TonB-dependent iron uptake pathway.

Accurate prediction of aviation safety levels is significant for the efficient early warning and prevention of incidents. However, the causal mechanism and temporal character of aviation accidents are complex and not fully understood, which increases the operation cost of accurate aviation safety prediction. This paper adopts an innovative statistical method involving a least absolute shrinkage and selection operator (LASSO) and long short-term memory (LSTM). We compiled and calculated 138 monthly aviation insecure events collected from the Aviation Safety Reporting System (ASRS) and took minor accidents as the predictor. Firstly, this paper introduced the group variables and the weight matrix into LASSO to realize the adaptive variable selection. Furthermore, it took the selected variable into multistep stacked LSTM (MSSLSTM) to predict the monthly accidents in 2020. Finally, the proposed method was compared with multiple existing variable selection and prediction methods. The results demonstrate that the RMSE (root mean square error) of the MSSLSTM is reduced by 41.98%, compared with the original model; on the other hand, the key variable selected by the adaptive spare group lasso (ADSGL) can reduce the elapsed time by 42.67% (13 s). This shows that aviation safety prediction based on ADSGL and MSSLSTM can improve the prediction efficiency of the model while keeping excellent generalization ability and robustness.

Prior knowledge alters perception already on early levels of processing. For instance, judging the display size of an object is affected by its familiar size. Using functional magnetic resonance imaging, we investigated the neural processes involved in resolving ambiguities between familiar object size and physical object size in 33 healthy human subjects. The familiar size was either small or large, and the object was displayed as either small or large. Thus, the size of the displayed object was either congruent or incongruent with its internally stored canonical size representation. Subjects were asked to indicate where the stimuli appeared on the screen as quickly and accurately as possible, thereby ensuring that differential activations cannot be ascribed to explicit object size judgments. Incongruent (relative to congruent) object displays were associated with enhanced activation of the right intraparietal sulcus (IPS). These data are consistent with but extend previous patient studies, which found the right parietal cortex involved in matching visual objects presented atypically to prototypical object representations, suggesting that the right IPS supports view normalization of objects. In a second experiment, using a parametric design, a region-of-interest analysis supported this notion by showing that increases in size mismatch between the displayed size of an object and its familiar viewing size were associated with an increased right IPS activation. We conclude that the right IPS performs view normalization of mismatched information about the internally stored prototypical size and the current viewing size of an object.

Recently, the homogenous flood generating mechanism assumption has become questionable due to changes in the underlying surface. In addition, flood is a multifaced natural phenomenon and should be characterized by both peak discharge and flood volume, especially for flood protection structures. Hence, in this study, data relating to the 55-year reservoir inflow, annual maximum flood peak (AMFP), and annual maximum flood volume (AMFV) for the Longmen Reservoir in North China have been utilized. The 1-day AMFV exhibits a significant correlation with AMFP. The extreme flood peak-volume pairs are then used to detect the heterogeneity and to perform nonstationary flood risk assessment using mixture distribution as the univariate marginal distribution. Moreover, a copula-based bivariate nonstationary flood frequency analysis is developed to investigate environmental effects on the dependence of flood peak and volume. The results indicate that the univariate nonstationary return period is between the joint OR and the AND return periods. The conditional probabilities of 1-day AMFV, when AMFP exceeds a certain threshold, are likely to be high, and the design flood values estimated by joint distribution are larger than the ones in the univariate nonstationary context. This study can provide useful information for engineers and decision-makers to improve reservoir flood control operations.

Illusory figures demonstrate the visual system’s ability to integrate separate parts into coherent, whole objects. The present study was performed to track the neuronal object construction process in human observers, by incrementally manipulating the grouping strength within a given configuration until the emergence of a whole-object representation. Two tasks were employed: First, in the spatial localization task, object completion could facilitate performance and was task-relevant, whereas it was irrelevant in the second, luminance discrimination task. Concurrent functional magnetic resonance imaging (fMRI) used spatial localizers to locate brain regions representing task-critical illusory-figure parts to investigate whether the step-wise object construction process would modulate neural activity in these localized brain regions. The results revealed that both V1 and the lateral occipital complex (LOC, with sub-regions LO1 and LO2) were involved in Kanizsa figure processing. However, completion-specific activations were found predominantly in LOC, where neural activity exhibited a modulation in accord with the configuration’s grouping strength, whether or not the configuration was relevant to performing the task at hand. Moreover, right LOC activations were confined to LO2 and responded primarily to surface and shape completions, whereas left LOC exhibited activations in both LO1 and LO2 and was related to encoding shape structures with more detail. Together, these results demonstrate that various grouping properties within a visual scene are integrated automatically in LOC, with sub-regions located in different hemispheres specializing in the component sub-processes that render completed objects.

The accurate design flood of hydraulic engineering is an important precondition to ensure the safety of residents, and the high precision estimation of flood frequency is a vital perquisite. The Xiangjiang River basin, which is the largest river in Hunan Province of China, is highly inclined to floods. This paper aims to investigate the annual maximum flood peak (AMFP) risk of Xiangjiang River basin under the climate context employing the Bayesian nonstationary time-varying moment models. Two climate covariates, i.e., the average June-July-August Artic Oscillation and sea level pressure in the Northwest Pacific Ocean, are selected and found to exhibit significant positive correlation with AMFP through a rigorous statistical analysis. The proposed models are tested with three cases, namely, stationary, linear-temporal and climate-based conditions. The results both indicate that the climate-informed model demonstrates the best performance as well as sufficiently explain the variability of extreme flood risk. The nonstationary return periods estimated by the expected number of exceedances method are larger than traditional ones built on the stationary assumption. In addition, the design flood could vary with the climate drivers which has great implication when applied in the context of climate change. This study suggests that nonstationary Bayesian modelling with climatic covariates could provide useful information for flood risk management.

To identify the major source of Salmonella contamination in a pig slaughterhouse, samples were collected from the clean and unclean area and Salmonella isolates were further typed. Carcasses entering the clean area showed a Salmonella contamination rate of 96.7% in the oral cavity and 55.0% in the rectum content samples. Evisceration seemed not to be critical as the contamination rate of the carcasses was similar before (16.7%) and after (18.3%) this slaughter step. In the unclean area, a limited number of oral cavity samples were positive after bleeding, while a dramatic increase of positives was observed after dehairing. Salmonella was detected in up to 0.01 mL of the recycled water collected from the dehairing machine. Genotyping of Salmonella isolates showed that similar pulsotypes were present in the oral cavity and recycled water. Based on these observations it can be concluded that the recycled water used in the dehairing machine was the major source for the carcass contamination in this slaughterhouse.

The pregnane X receptor (PXR) has been well-established as a critical mediator in regulating important drug metabolizing enzymes and transporter proteins, including cytochrome P450 3A4 (CYP3A4) and P-glycoprotein (P-gp). Previous studies identified that PXR served as a molecular target of SUMOylation. However, the impact of SUMOylation of PXR on its transcriptional activity in regulating the expression/activity of the target genes is poorly investigated. In the current study, we established cell-based models of SUMOylated PXR in LS174T cells to investigate the impact of SUMOylation of PXR on regulating the expression/activity of CYP3A4 and P-gp. Our results revealed that rifampicin-induced PXR transactivation of the CYP3A4 and P-gp promoter was suppressed by SUMOylation of PXR in reporter gene assay. The mRNA and protein expression of CYP3A4 and P-gp was also suppressed. Moreover, CYP3A4 enzymatic assay and Rho123 intracellular assay revealed that rifampicin-induced CYP3A4 and P-gp activity was also suppressed by SUMOylated PXR. Our data collectively indicated for the first time that SUMOylation of PXR exerts suppressive effect on rifampicin-induced expression and activity of CYP3A4 and P-gp, which suggest that alteration in the SUMOylation status of PXR will be expected to affect the CYP3A4 mediated drug metabolism and P-gp mediated drug transport.

Background/Aims: To explore whether Nrf2 was associated with drug-resistance in cisplatin resistant A549 (A549/DDP) cells, and if cryptotanshinone (CTS), one of the bioactive compounds isolated from the roots of Salvia miltiorrhiza Bunge (Danshen), could enhance the sensitivity in A549/DDP cells towards cisplatin. Methods: A549 and A549/DDP cells were subjected to various treatments, and then Sulforhodamine B (SRB) assay, flow cytometry analysis and western immunoblotting analysis were applied to determine IC50, apoptotic status and expressions of Nrf2 and its downstream genes. Results: The endogenous expression levels of Nrf2 as well as its target genes including GCLC, GCLM, HO-1, NQO1 and MRP1 were much higher in A549/DDP cells than those of A549 cells and the susceptibility of A549/DDP cells to cisplatin was partially restored by silencing Nrf2. The combination of CTS and cisplatin led to cell death and apoptosis through sensitizing A549/DDP cells towards cisplatin compared with cisplatin mono-treatment, however, this reversal role could be abolished by Nrf2 knockdown. Specifically, CTS obviously diminished Nrf2 expression, thus contributing to the decrease of Nrf2-target genes expression levels. Meanwhile, we also discovered that CTS triggered several other signals involving in chemoresistance such as MAPKs, Akt and STAT3 pathway. Conclusion: Our data indicated CTS may be developed as a potential sensitizer cooperating with anticancer drugs to combat chemoresistant carcinoma through the inhibition of the Nrf2 pathway.

Background: Aberrant expression of Nicotinamide N-methyltransferase (NNMT) has been reported in pancreatic cancer. However, the role of NNMT in pancreatic cancer development remains elusive. Therefore, the present study was to investigate the impact of NNMT on pancreatic cancer cell proliferation, metastatic potential and survival under metabolic stress. Methods: Pancreatic cancer cell line PANC-1 was transfected with NNMT expression plasmid or small interfering RNA of NNMT to overexpress or knockdown intracellular NNMT expression, respectively. Rate of cell proliferation was monitored. Transwell migration and matrigel invasion assays were conducted to assess cell migration and invasion capacity. Resistance to glucose deprivation, sensitivity to glycolytic inhibition, mitochondrial inhibtion and resistance to rapamycin were examined to evaluate cell survival under metabolic stress. Results: NNMT silencing markedly reduced cell proliferation, whereas NNMT overexpression promoted cell growth moderately. Knocking down NNMT also significantly suppressed the migration and invasion capacities of PANC-1 cells. Conversely, NNMT upregulation enhanced cell migration and invasion capacities. In addition, NNMT knockdown cells were much less resistant to glucose deprivation and rapamycin as well as glycolytic inhibitor 2-deoxyglucose whereas NNMT-expressing cells showed opposite effects although the effects were not so striking. Conclusions: These data sugguest that NNMT plays an important role in PANC-1 cell proliferation, metastatic potential and survival under metabolic stress.

This study focused on investigating the pathogenesis seen in specific-pathogen-free (SPF) rabbits following infection with a homologous rabbit HEV isolate (CHN-BJ-rb14) and comparing it to that seen following infection with a heterologous swine genotype 4 HEV isolate (CHN-XJ-SW13). Three of the four animals inoculated with the homologous rabbit HEV became infected, exhibiting an intermittent viremia, obvious fluctuations of liver function biomarkers alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and persistent fecal virus shedding throughout the nine month study. In addition, liver histopathology showed both chronic inflammation and some degree of fibrosis. Both positive and negative-stranded HEV RNA and HEV antigen expression were detected in liver, brain, stomach, duodenum and kidney from the necropsied rabbits. Inflammation of extrahepatic tissue (duodenum and kidney) was also observed. Three of the four rabbits inoculated with the heterologous genotype 4 swine HEV also became infected, showing similar levels of anti-HEV antibody to that generated following infection with the homologous virus isolate. The duration of both viremia and fecal shedding of virus was however shorter following infection with the heterologous virus and there was no significant elevation of liver function biomarkers. These results suggest that rabbit HEV infection may cause more severe hepatitis and prolong the course of the disease, with a possible chronic trend of hepatitis in SPF rabbits.