Tandem gene duplication selected by activation of horizontally transferred gene in bacteria.

Horizontal gene transfer occurs frequently in bacteria, but the mechanism driving activation and optimization of the expression of horizontally transferred genes (HTGs) in new recipient strains is not clear. Our previous study found that spontaneous tandem DNA duplication resulted in rapid activation of HTGs. Here, we took advantage of this finding to develop a novel technique for tandem gene duplication, named tandem gene duplication selected by activation of horizontally transferred gene in bacteria (TDAH), in which tandem duplication was selected by the activation of horizontally transferred selectable marker gene. TDAH construction does not contain any reported functional elements based on homologous or site-specific recombination and DNA amplification. TDAH only contains an essential selectable marker for copy number selection and 9-bp-microhomology border sequences for precise illegitimate recombination. One transformation and 3 days were enough to produce a high-copy strain, so its procedure is simple and fast. Without subsequent knockout of the endogenous recombination system, TDAH could also generate the relatively stable high-copy tandem duplication for plasmid-carried and genome-integrated DNA. TDAH also showed an excellent capacity for increase gene expression and worked well in different industrial bacteria. We also applied TDAH to select the optimal high copy number of ribA for vitamin B2 production in E. coli; the yield was improved by 3.5 times and remained stable even after 12 subcultures. TDAH is a useful tool for recombinant protein production and expression optimization of biosynthetic pathways. KEY POINTS: • We develop a novel and efficient technique (TDAH) for tandem gene duplication in bacterium. TDAH is based on the mechanism of HTG rapid activation. TDAH does not contain any reported functional elements based on homologous recombination and DNA amplification. TDAH only contains an essential selectable marker for copy number selection, so its construction and procedure are very simple and fast. • TDAH is the first reported selected and stable tandem-gene-duplication technique in which the selected high-copy plasmid-carried and genome-integrated DNA could remain stable without the subsequent knockout of recombination system. • TDAH showed an excellent capacity for regulating gene expression and worked well in different industrial bacteria, indicating it is a useful tool for recombinant protein production and expression optimization of biosynthetic pathways. • TDAH was applied to select the optimal high copy number of ribA for vitamin B2 production in E. coli; the yield was improved by 3.5-fold and remained stable even after 12 subcultures.

Effects of Artificial Sugar Supplementation on the Composition and Nutritional Potency of Honey from Apis cerana.

In the global apiculture industry, reward feeding and supplementary feeding are essential for maintaining bee colonies. Beekeepers provide artificial supplements to their colonies, typically in the form of either a honey-water solution or sugar syrup. Owing to cost considerations associated with beekeeping, most beekeepers opt for sugar syrup. However, the effects of different types of artificial sugar supplements on bee colonies and their subsequent impact on honey composition remain unclear. To address this gap, this study compared the chemical composition, antioxidant capacity, and nutritional potency of three types of honey: honey derived from colonies fed sugar syrup (sugar-based product, SP) or a honey-water solution (honey-sourced honey, HH) and naturally sourced honey (flower-sourced honey, FH), which served as the control. The results revealed that FH outperformed HH and SP in terms of total acidity, sugar content, total protein content, and antioxidant capacity, and HH outperformed SP. Regarding nutritional efficacy, including the lifespan and learning and memory capabilities of worker bees, FH exhibited the best outcomes, with no significant differences observed between HH and SP. This study underscores the importance of sugar source selection in influencing honey quality and emphasizes the potential consequences of substituting honey with sugar syrup in traditional apiculture practices.

Synergistic resistance of honeybee (Apis mellifera) and their gut microorganisms to fluvalinate stress.

Fluvalinate is widely used in the control of Varroa destructor, but its residues in colonies threaten honeybees. The effect of fluvalinate-induced dysbiosis on honeybee-related gene expression and the gut microenvironment of honeybees has not yet been fully elucidated. In this study, two-day-old larvae to seven-day-old adult worker bees were continuously fed different amounts of fluvalinate-sucrose solutions (0, 0.5, 5, and 50 mg/kg), after which the expression levels of two immune-related genes (Hymenoptaecin and Defensin1) and three detoxication-related genes (GSTS3, CAT, and CYP450) in worker bees (1, 7, and 20 days old) were measured. The effect of fluvalinate on the gut microbes of worker bees at seven days old also was explored using 16S rRNA Illumina deep sequencing. The results showed that exposure of honeybees to the insecticide fluvalinate affected their gene expression and gut microbial composition. As the age of honeybees increased, the effect of fluvalinate on the expression of Hymenoptaecin, CYP450, and CAT decreased, and the abundance of honeybee gut bacteria was affected by increasing the fluvalinate concentration. These findings provide insights into the synergistic defense of honeybee hosts against exogenous stresses in conjunction with honeybee gut microbes.

Acetylation of transcription factor BpTCP20 by acetyltransferase BpPDCE23 modulates salt tolerance in birch.

Teosinte branched 1/Cycloidea/Proliferating cell factor (TCP) transcription factors function in abiotic stress responses. However, how TCPs confer salt tolerance is unclear. Here, we characterized a TCP transcription factor, BpTCP20, that responds to salt stress in birch (Betula platyphylla Suk). Plants overexpressing BpTCP20 displayed increased salt tolerance, and Bptcp20 knockout mutants displayed reduced salt tolerance relative to the wild-type (WT) birch. BpTCP20 conferred salt tolerance by mediating stomatal closure and reducing reactive oxygen species accumulation. Chromatin immunoprecipitation sequencing showed that BpTCP20 binds to NeuroD1, T-box, and two unknown elements (termed TBS1 and TBS2) to regulate target genes. In birch, salt stress led to acetylation of BpTCP20 acetylation at lysine 259. A mutated BpTCP20 variant (abolished for acetylation, termed BpTCP20259) was overexpressed in birch, which led to decreased salt tolerance compared with plants overexpressing BpTCP20. However, BpTCP20259-overexpressing plants still displayed increased salt tolerance relative to untransformed wild-type plants. BpTCP20259 showed reduced binding to the promoters of target genes and decreased target gene activation, leading to decreased salt tolerance. In addition, we identified dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex (BpPDCE23), an acetyltransferase that interacts with and acetylates BpTCP20 to enhance its binding to DNA motifs. Together, these results suggest that BpTCP20 is a transcriptional regulator of salt tolerance, whose activity is modulated by BpPDCE23-mediated acetylation.

Comprehensive bioinformatics analysis reveals the role of cuproptosis-related gene Ube2d3 in myocardial infarction.

Background: Myocardial infarction (MI) caused by severe coronary artery disease has high incidence and mortality rates, making its prevention and treatment a central and challenging aspect of clinical work for cardiovascular practitioners. Recently, researchers have turned their attention to a novel mechanism of cell death caused by Cu2+, cuproptosis. Methods: This study integrated data from three MI-related bulk datasets downloaded from the Gene Expression Omnibus (GEO) database, and identified 16 differentially expressed genes (DEGs) related to cuproptosis by taking intersection of the 6378 DEGs obtained by differential analysis with 49 cuproptosis-related genes. Four hub genes, Dbt, Dlat, Ube2d1 and Ube2d3, were screened out through random forest analysis and Lasso analysis. In the disease group, Dbt, Dlat, and Ube2d1 showed low expression, while Ube2d3 exhibited high expression. Results: Focusing on Ube2d3 for subsequent functional studies, we confirmed its high expression in the MI group through qRT-PCR and Western Blot detection after successful construction of a MI mouse model by left anterior descending (LAD) coronary artery ligation, and further clarified the correlation of cuproptosis with MI development by detecting the levels of cuproptosis-related proteins. Moreover, through in vitro experiments, Ube2d3 was confirmed to be highly expressed in oxygen-glucose deprivation (OGD)-treated cardiomyocytes AC16. In order to further clarify the role of Ube2d3, we knocked down Ube2d3 expression in OGD-treated AC16 cells, and confirmed Ube2d3's promoting role in the hypoxia damage of AC16 cells by inducing cuproptosis, as evidenced by the detection of MTT, TUNEL, LDH release and cuproptosis-related proteins. Conclusion: In summary, our findings indicate that Ube2d3 regulates cuproptosis to affect the progression of MI.

Plasmon-Stimulated Colorimetry Biosensor Array for the Identification of Multiple Metabolites.

Rationally designing highly catalytic and stable nanozymes for metabolite monitoring is of great importance because of their huge potential in early disease diagnosis. Herein, a novel nanozyme based on hierarchically structured CuS/ZnS with a highly efficient peroxidase (POD)-mimic capability was developed and synthesized for multiple metabolite determination and recognition via the plasmon-stimulated biosensor array strategy. The designed nanozyme can simultaneously harvest plasmon triggered hot electron-hole pairs and generate photothermal properties, leading to a sharply boosted POD-mimic capability under 808 nm laser irradiation. Interestingly, because of the interaction diversity of the metabolite with POD-like nanomaterials, the unique inhibitory effect of metabolites on the POD-mimic activity could be the signal response as the differentiation. Thus, utilizing TMB as a typical chromogenic substrate in the addition of H2O2, the designed colorimetric biosensor array can produce diverse fingerprints for the three vital metabolisms (cysteine (Cys), ascorbic acid (AA), and glutathione (GSH)), which can be precisely identified by principal component analysis (PCA). Notably, a distinct fingerprint of a single metabolite with different levels and metabolite mixtures is also achieved with a detection limit of 1 µM. Most importantly, cell lysis could be effectively discriminated by the biosensor assay, implying its great potential in clinical diagnosis.

Rapid Analysis of Estrogens in Meat Samples by High Performance Liquid Chromatography with Fluorescence Detection.

A novel reagent named 4-(N-methyl-1,3-dioxo-benzoisoquinolin-6-yl-oxy)benzene sulfonyl chloride (MBIOBS-Cl) for the determination of estrogens in food samples by high-performance liquid chromatography (HPLC) with fluorescence detection has been developed. Estrogens could be easily labeled by MBIOBS-Cl in Na2CO3-NaHCO3 buffer solution at pH 10.0. The complete labeling reaction for estrogens could be accomplished within five minutes, the corresponding derivatives exhibited strong fluorescence with the maximum excitation and emission wavelengths at 249 nm and 443 nm, respectively. The derivatization conditions, such as the molar ratio of reagent to estrogens, derivatization time, pH, temperature, and buffers were optimized. Derivatives were sufficiently stable to be efficiently analyzed by HPLC with a reversed-phase Agilent ZORBAX 300SB-C18 column with a good baseline resolution. Excellent linear correlations were obtained for all estrogen derivatives with correlation coefficients greater than 0.9998. Ultrasonic-Assisted extraction was used to optimize the extraction of estrogens from meat samples with a recovery higher than 82%. The detection limits (LOD, S/N = 3) of the method ranged from 0.95 to 3.3 µg· kg-1. The established method, which is fast, simple, inexpensive, and environment friendly, can be successfully applied for the detection of four steroidal estrogens from meat samples with little matrix interference.

Fast clustering algorithm based on MST of representative points.

Minimum spanning tree (MST)-based clustering algorithms are widely used to detect clusters with diverse densities and irregular shapes. However, most algorithms require the entire dataset to construct an MST, which leads to significant computational overhead. To alleviate this issue, our proposed algorithm R-MST utilizes representative points instead of all sample points for constructing MST. Additionally, based on the density and nearest neighbor distance, we improved the representative point selection strategy to enhance the uniform distribution of representative points in sparse areas, enabling the algorithm to perform well on datasets with varying densities. Furthermore, traditional methods for eliminating inconsistent edges generally require prior knowledge about the number of clusters, which is not always readily available in practical applications. Therefore, we propose an adaptive method that employs mutual neighbors to identify inconsistent edges and determine the optimal number of clusters automatically. The experimental results indicate that the R-MST algorithm not only improves the efficiency of clustering but also enhances its accuracy.

Photo-Controlled Nano-Supramolecular Size and Reversible Luminescent Behaviors Based on Cucurbit[7]uril Cascaded Assembly.

Supramolecular luminescent material with switchable behavior and photo-induced aggregation with emission enhancement is a current research hot spot. Herein, a size-tunable nano-supramolecular assembly with reversible photoluminescent behavior was constructed by noncovalent polymerization of diarylethene-bridged bis(coumarin) derivative (DAE-CO), cucurbit[7]uril (CB[7]), and ß-cyclodextrin-grafted hyaluronic acid (HACD). Benefiting from the macrocyclic confinement effect, the guest molecule DAE-CO was included into the cavity of CB[7] to give enhanced fluorescence emission of the resulting DAE-CO⊂CB[7]2 with longer lifetime at 432 nm to 1.43 ns, thereby further enhancing fluorescence output and lifetime (1.46 ns) when further assembled with HACD, compared with the free DAE-CO (0.95 ns). In addition, DAE-CO, DAE-CO⊂CB[7]2, and DAE-CO⊂CB[7]2&HACD all possessed characteristics of aggregation-induced emission and reversible photo-switched structural interconversion, exhibiting an obvious photophysical activation phenomenon of self-aggregation into larger nanoparticles with increase in fluorescence emission intensity, lifetime, and size after irradiation, which could be increased step by step with the alternating irradiation of 254 nm (5 min) or >600 nm (30 s) repeated 7 times. These supramolecular assemblies were successfully used in the tumor cells' targeted imaging and anti-counterfeiting because of the capability of HACD for recognizing specific receptors overexpressed on the surface of tumor cells and the excellent photo-regulated switch ability of DAE-CO, providing an approach of constructing photo-induced emission-enhanced luminescent materials.

A novel density peaks clustering algorithm for automatic selection of clustering centers based on K-nearest neighbors.

The density peak clustering algorithm (DPC) requires manual determination of cluster centers, and poor performance on complex datasets with varying densities or non-convexity. Hence, a novel density peak clustering algorithm is proposed for the automatic selection of clustering centers based on K-nearest neighbors (AKDPC). First, the AKDPC classifies samples according to their mutual K-nearest neighbor values into core and non-core points. Second, the AKDPC uses the average distance of K nearest neighbors of a sample as its density. The smaller the average distance is, the higher the density. Subsequently, it selects the highest density sample among all unclassified core points as a center of the new cluster, and the core points that satisfy the merging condition are added to the cluster until no core points satisfy the condition. Afterwards, the above steps are repeated to complete the clustering of all core points. Lastly, the AKDPC labels the unclassified non-core points similar to the nearest points that have been classified. In addition, to prove the validity of AKDPC, experiments on manual and real datasets are conducted. By comparing the AKDPC with classical clustering algorithms and excellent DPC-variants, this paper demonstrates that AKDPC presents higher accuracy.

Honey bee maternal effects improve worker performance and reproductive ability in offspring.

Maternal effects are an evolutionary strategy used to improve offspring quality. In an example of maternal effects in honey bees (Apis mellifera), mother queens produce larger eggs in queen cells than in worker cells in order to breed better daughter queens. In our current study, morphological indexes, reproductive tissues, and the egg-laying ability of newly reared queens reared with eggs laid in queen cells (QE), eggs laid in worker cells (WE), and 2-day-old larvae in worker cells (2L) were evaluated. In addition, morphological indexes of offspring queens and working performance of offspring workers were examined. The thorax weight, number of ovarioles, egg length, and number of laid eggs and capped broods of QE were significantly higher than those of WE and 2L, indicating that the reproductive capacity of QE group was better than that of other groups. Furthermore, offspring queens from QE had larger thorax weights and sizes than those from the other two groups. Offspring worker bees from QE also had larger body sizes and greater pollen-collecting and royal jelly-producing abilities than those of other two groups. These results demonstrate that honey bees display profound maternal effects on queen quality that can be transmitted across generations. These findings provide a basis for improving queen quality, with implications in apicultural and agricultural production.

Metabolic profiling of Apis mellifera larvae treated with sublethal acetamiprid doses.

Acetamiprid is a neonicotinoid insecticide used in crop protection worldwide. Such widespread application can pose risks to pollinator insects, particularly to honeybees (Apis mellifera); therefore, the evaluation of the harmful effects of acetamiprid is necessary. Recent studies report behavior and gene expression dysfunction in honeybees, related to acetamiprid contamination. However, most studies do not consider potential metabolism disorders. To examine the effects of sublethal acetamiprid doses on the hemolymph metabolism of honeybees, worker bee larvae(2 days old) were fed with sucrose water containing different concentrations of acetamiprid (0, 5, and 25 mg/L) until capped (6 days old). The hemolymph (200 µL) of freshly capped larvae was collected for liquid chromatography-mass spectrometry (LC-MS). Overall, increasing acetamiprid exposure induced greater metabolic variations in worker bee larvae(treated groups compared to untreated). In the positive ion mode, 36 common differential metabolites in the acetamiprid-treated groups were screened from the identified differential metabolites. Of these, 19 metabolites were upregulated, and 17 were downregulated. 10 common differential metabolites were screened in the negative ion mode. 3 metabolites were upregulated, and 7 metabolites were downregulated. These common metabolites included traumatic acid, indole etc. These commonly differentiated metabolites were classified as compounds with biological roles, lipids, and phytochemical compounds, and others. The metabolic pathways of common differentiated metabolites with significant differences (P < 0.05) included the metabolism of tryptophan, purines, phenylalanine, etc. As the concentration of acetamiprid increased, the content of traumatic acid increased, the content of tryptophan metabolite l-kynurenine and indole decreased, and the content of lipids also decreased. Our results revealed that the damage to honeybee larvae increased when the acetamiprid solution formulations residue in their food had a concentration higher than 5 mg/L, causing metabolic disorders in various substances in larvae. Analysis of these metabolic processes can provide a theoretical basis for further research on the metabolism of acetamiprid-treated honeybees and elucidate the detoxification mechanisms.

Wnt signaling regulates MFSD2A-dependent drug delivery through endothelial transcytosis in glioma.

BACKGROUND: Systemic delivery of anti-tumor therapeutic agents to brain tumors is thwarted by the blood-brain barrier (BBB), an organotypic specialization of brain endothelial cells (ECs). A failure of pharmacological compounds to cross BBB is one culprit for the dismal prognosis of glioblastoma (GBM) patients. Identification of novel vascular targets to overcome the challenges posed by the BBB in tumors for GBM treatment is urgently needed. METHODS: Temozolomide (TMZ) delivery was investigated in CT2A and PDGFB-driven RCAS/tv-a orthotopic glioma models. Transcriptome analysis was performed on ECs from murine gliomas. Mfsd2a deficient, Cav1 deficient, and Mfsd2a EC-specific inducible mice were developed to study the underlying molecular mechanisms. RESULTS: We demonstrated that inhibiting Wnt signaling by LGK974 could increase TMZ delivery and sensitize glioma to chemotherapy in both murine glioma models. Transcriptome analysis of ECs from murine gliomas revealed that Wnt signaling inhibition enhanced vascular transcytosis as indicated by the upregulation of PLVAP and downregulation of MFSD2A. Mfsd2a deficiency in mice enhances TMZ delivery in tumors, whereas constitutive expression of Mfsd2a in ECs suppresses the enhanced TMZ delivery induced by Wnt pathway inhibition in murine glioma. In addition, Wnt signaling inhibition enhanced caveolin-1 (Cav1)-positive caveolae-mediated transcytosis in tumor ECs. Moreover, Wnt signaling inhibitor or Mfsd2a deficiency fails to enhance TMZ penetration in tumors from Cav1-deficient mice. CONCLUSIONS: These results demonstrated that Wnt signaling regulates MFSD2A-dependent TMZ delivery through a caveolae-mediated EC transcytosis pathway. Our findings identify Wnt signaling as a promising therapeutic target to improve drug delivery for GBM treatment.

Supplementing Diets with Agriophyllum squarrosum Reduced Blood Lipids, Enhanced Immunity and Anti-Inflammatory Capacities, and Mediated Lipid Metabolism in Tan Lambs.

Agriophyllum squarrosum (sand rice), a widespread desert plant, possesses anti-hyperglycemic and anti-inflammatory properties, and has been used in traditional Chinese medicine for many years. However, its effects on ruminants are unknown. To fill this gap, we examined the effects of A. squarrosum on the immune and anti-inflammatory responses of lambs. A total of 23, 6-month-old Tan ewe-lambs (27.6 ± 0.47 kg) were divided into four groups and offered a basic diet (C­control), or a diet that contained 10%, 20%, or 30% A. squarrosum, on a dry matter basis, for 128 days. Serum concentrations of total cholesterol were lower (p = 0.004) in the 30% supplemented lambs than controls, while concentrations of high-density lipoprotein cholesterol were lower (p = 0.006) in the 10% and 20%, but not in 30% supplemented lambs than controls. Serum-cortisol concentrations were lower (p = 0.012) in the 30% supplemented lambs and free fatty acid concentrations were higher in the 10% and 20% supplemented lambs than in control lambs (p < 0.001). Supplementation with A. squarrosum decreased (p < 0.05) the area of adipocytes in subcutaneous adipose tissue, but there was no difference between the 20% and 30% diets. Conversely, the area in visceral adipose tissue (VAT) increased (p < 0.05), especially for the 10% and 20% supplemented diets. Supplementation with A. squarrosum also enriched immune and anti-inflammatory related and lipid and glucose-metabolic pathways and associated differentially expressed gene expressions in adipose tissue. A total of 10 differential triacylglycerol, 34 differential phosphatidylcholines and seven differential phosphatidylethanolamines decreased in the diet with 30% supplementation, when compared to the other diets. Finally, adipocyte-differentiation genes, and immune and inflammatory response-related gene expression levels decreased in lamb adipocytes cultured with an aqueous A. squarrosum extract. In conclusion, supplementing lamb diets with A. squarrosum reduced blood lipids, enhanced immunity and anti-inflammatory capacities, and mediated lipid metabolism in adipose tissue and adipocytes of Tan lambs. A level of approximately 10% is recommended, but further research is required to determine the precise optimal level.

Association of coal mine dust lung disease with Nodular thyroid disease in coal miners: A retrospective observational study in China.

Background: Coal dust is a major risk factor for the occupational health of coal miners, and underground workers with coal mine dust lung disease (Coal miners with coal mine dust lung disease (CMDLD) may have a higher risk of developing Nodular thyroid disease (NTD). The aim of this study was to investigate the relationship between coal mine dust lung disease and the development of Nodular thyroid disease in coal miners. Methods: This was a clinical retrospective observational study that included 955 male coal miners from 31 different coal mining companies in Huainan, Anhui Province, China, who were examined in April 2021 at the Huainan Occupational Disease Prevention and Control Hospital to collect all their clinical physical examination data, including general conditions, laboratory test indices and imaging indices. Based on the presence or absence of Nodular thyroid disease, 429 cases with Nodular thyroid disease were classified as the diseased group and 526 cases without Nodular thyroid disease were classified as the control group. Logistic regression was used to analyse the correlation between the occurrence of Nodular thyroid disease in coal miners, and further single- and multi-factor logistic regression was used to screen the risk exposure factors for Nodular thyroid disease in coal miners. Results: Age, coal mine dust lung disease (CMDLD), red blood cells (RBC), mean red blood cell volume (MCV), albumin (ALB), albumin/globulin (A/G), indirect bilirubin (IBIL), globulin (GLOB), total bilirubin (TBil) and myeloperoxidase (MPO) were associated with the development of Nodular thyroid disease in coal miners (p < 0.05) The results of univariate and multifactorial logistic regression analysis showed that CMDLD (OR:4.5,95%CI:2.79-7.51) had the highest OR and CMDLD was the strongest independent risk exposure factor for the development of Nodular thyroid disease in coal miners. Conclusions: There is a strong correlation between coal mine dust lung disease and Nodular thyroid disease in underground coal miners, and clinicians need to be highly aware of the high risk of NTD in coal miners with CMDLD and adopt individualized clinical prevention strategies.

High-Quality Queens Produce High-Quality Offspring Queens.

Honey bees, rather than rear queens with eggs and larvae from worker cells, prefer to rear new queens with eggs form queen cells, if available. This may be a result of long-term evolutionary process for honey bee colonies. However, the exact mechanism of this phenomenon is unclear. In this study, queens were reared with eggs from queen cells (F1-QE), eggs from worker cells (F1-WE), and two-day-old larvae from worker cells (F1-2L). Physiological indexes and the expression of the development-related genes ((Hexamerin (Hex110, Hex70b), Transferrin (Trf), and Vitellogenin (Vg)) of reared F1 generation queens were measured and compared. Furthermore, F2 generation queens were reared with one-day-old larvae from F1 queens, and the weight and ovariole count of reared F2 generation daughter queens were examined. Meanwhile, the expression of the development- and reproduction-related genes (Hex110, Hex70b, Trf, Vg, and Juvenile Hormone (Jh)) and immune detoxication-related genes (Hymenoptaecin, Abeacin, and CytP450) of reared F2 queens were further explored. We found that the F1-QE queens had the highest physiological indexes and higher Hex110 and Trf expression levels, while no significant difference was found in the expression of Hex70b and Vg among the three groups of F1 queens. In addition, the reared queens of F2-QE had the highest quality, with the highest development, reproduction, immune-detoxication genes' expression levels. Our results revealed that the quality of reared offspring queens from high-quality mother queens was also high. These findings inform methods for rearing high-quality queens and highlight that a high-quality queen is essential for offspring colony growth and survival.

circ_0044516 functions in the progression of gastric cancer by modulating MicroRNA-149-5p/HuR axis.

Circular RNAs (circRNAs) have emerged as a multifunctional class of RNAs, while there is limited knowledge on their functions in the development of cancers. Herein, we performed the current study to probe into the regulatory mechanism of circ_0044516 in malignant behaviors of gastric cancer (GC) cells with the involvement of microRNA (miR)-149-5p/human antigen R (HuR) axis. Firstly, the expression levels of circ_0044516 in GC cell lines and normal gastric mucosal epithelial cells were determined by qRT-PCR, and GC cell lines with the highest expression of circ_0044516 were screened for further cell experiments. Subsequently, the biological functions of silenced circ_0044516 in GC were identified by CCK-8, colony formation, and transwell assays. Xenograft mouse models were established for in vivo verification. Furthermore, luciferase reporter, RIP, RNA pull-down assay and rescue experiments were performed to explore the sponge regulatory mechanism of circ_0044516. circ_0044516 was suggested to be highly expressed in GC cell lines, and circ_0044516 could promote GC cell proliferation, migration and invasion, as well as in vivo tumor growth. In addition, silenced circ-0044516 reversed the promotive roles in cell viability caused by overexpressed HuR. Furthermore, circ_0044516 mainly localized in the cytoplasm, which may act as a miR-149-5p sponge to modulate HuR expression, thereby playing an essential role in GC development. This study suggests that circ_0044516 may promote HuR expression through sponging miR-149-5p, thereby playing a part in GC progression.

School victimization and Internet addiction among Chinese adolescents: The mediating roles of life satisfaction and loneliness.

The present study investigated the possibility of life satisfaction and loneliness mediating the link between school victimization and Internet addiction. A total of 3,363 middle/high school students (45% males; Mage = 15.67 years old, SD = 1.58) completed a series of self-report questionnaires, which included school victimization, life satisfaction, loneliness, and Internet addiction. The findings demonstrated a positive relationship between school victimization and Internet addiction. In addition, life satisfaction and loneliness mediated the link between school victimization and Internet addiction. Overall, these findings contribute to a better understanding of the association between school victimization and Internet addiction. They also extended the GST, providing suggestions for preventing and managing adolescents' Internet addiction.

Development and validation of the Emotional Intelligence Test for Adolescents in a Chinese sample.

Emotional intelligence, or the ability to perceive, understand, and regulate emotions, has been identified as a protective factor for one's adaptation. Measuring emotional intelligence using performance-based approach is thought to be objective and effective. However, performance-based emotional intelligence instruments for non-Western adolescents are rarely available. To facilitate research on emotional intelligence, the present study developed and validated the emotional intelligence test for adolescents (A-EIT) using a Chinese sample. Study 1 (N = 1,536) showed that emotional intelligence consists of three subabilities (i.e., emotion perception, emotion understanding, and emotion regulation) and identified eligible items for each subtest through exploratory factor analysis (EFA). Study 2 (N = 2,568) confirmed the three-factor structure and its measurement invariance across gender and age groups by (multiple-group) confirmatory factor analyses. Both the full test and its three subtests showed acceptable internal consistencies, and their scores increased with age and showed female advantage. Also, the A-EIT scores were significantly correlated with indicators related to emotional and cognitive skills (convergent validity) and were independent of personality (discriminant validity). Additionally, higher scores on the A-EIT were related to better intrapersonal, interpersonal, and academic adaption (criterion validity), as expected. Study 3 (N = 163) provided further convergent validity evidence for each subtest by using validity tools not based on an emotional intelligence framework. Additionally, the A-EIT displayed satisfactory test-retest reliability. Generally, the A-EIT appears to be a valid and reliable instrument to assess adolescents' emotion intelligence, especially those with relatively low level. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Selective Chemical Labeling and Sequencing of 5-Hydroxymethylcytosine in DNA at Single-Base Resolution.

5-Hydroxymethylcytosine (5hmC), the oxidative product of 5-methylcytosine (5mC) catalyzed by ten-eleven translocation enzymes, plays an important role in many biological processes as an epigenetic mediator. Prior studies have shown that 5hmC can be selectively labeled with chemically modified glucose moieties and enriched using click chemistry with biotin affinity approaches. Besides, DNA deaminases of the AID/APOBEC family can discriminate modified 5hmC bases from cytosine (C) or 5mC. Herein, we developed a method based on embryonic stem cell (ESC) whole-genome analysis, which could enrich 5hmC-containing DNA by selective chemical labeling and locate 5hmC sites at single-base resolution with enzyme-based deamination. The combination experimental design is an extension of previous methods, and we hope that this cost-effective single-base resolution 5hmC sequencing method could be used to promote the mechanism and diagnosis research of 5hmC.