Epithelial to mesenchymal transition during mammalian neural crest cell delamination.

Epithelial to mesenchymal transition (EMT) is a well-defined cellular process that was discovered in chicken embryos and described as "epithelial to mesenchymal transformation" [1]. During EMT, epithelial cells lose their epithelial features and acquire mesenchymal character with migratory potential. EMT has subsequently been shown to be essential for both developmental and pathological processes including embryo morphogenesis, wound healing, tissue fibrosis and cancer [2]. During the past 5 years, interest and study of EMT especially in cancer biology have increased exponentially due to the implied role of EMT in multiple aspects of malignancy such as cell invasion, survival, stemness, metastasis, therapeutic resistance and tumor heterogeneity [3]. Since the process of EMT in embryogenesis and cancer progression shares similar phenotypic changes, core transcription factors and molecular mechanisms, it has been proposed that the initiation and development of carcinoma could be attributed to abnormal activation of EMT factors usually required for normal embryo development. Therefore, developmental EMT mechanisms, whose timing, location, and tissue origin are strictly regulated, could prove useful for uncovering new insights into the phenotypic changes and corresponding gene regulatory control of EMT under pathological conditions. In this review, we initially provide an overview of the phenotypic and molecular mechanisms involved in EMT and discuss the newly emerging concept of epithelial to mesenchymal plasticity (EMP). Then we focus on our current knowledge of a classic developmental EMT event, neural crest cell (NCC) delamination, highlighting key differences in our understanding of NCC EMT between mammalian and non-mammalian species. Lastly, we highlight available tools and future directions to advance our understanding of mammalian NCC EMT.

Dynamic regulation and requirement for ribosomal RNA transcription during mammalian development.

Ribosomal RNA (rRNA) transcription by RNA polymerase I (Pol I) is a critical rate-limiting step in ribosome biogenesis, which is essential for cell survival. Despite its global function, disruptions in ribosome biogenesis cause tissue-specific birth defects called ribosomopathies, which frequently affect craniofacial development. Here, we describe a cellular and molecular mechanism underlying the susceptibility of craniofacial development to disruptions in Pol I transcription. We show that Pol I subunits are highly expressed in the neuroepithelium and neural crest cells (NCCs), which generate most of the craniofacial skeleton. High expression of Pol I subunits sustains elevated rRNA transcription in NCC progenitors, which supports their high tissue-specific levels of protein translation, but also makes NCCs particularly sensitive to rRNA synthesis defects. Consistent with this model, NCC-specific deletion of Pol I subunits Polr1a, Polr1c, and associated factor Tcof1 in mice cell-autonomously diminishes rRNA synthesis, which leads to p53 protein accumulation, resulting in NCC apoptosis and craniofacial anomalies. Furthermore, compound mutations in Pol I subunits and associated factors specifically exacerbate the craniofacial anomalies characteristic of the ribosomopathies Treacher Collins syndrome and Acrofacial Dysostosis-Cincinnati type. Mechanistically, we demonstrate that diminished rRNA synthesis causes an imbalance between rRNA and ribosomal proteins. This leads to increased binding of ribosomal proteins Rpl5 and Rpl11 to Mdm2 and concomitantly diminished binding between Mdm2 and p53. Altogether, our results demonstrate a dynamic spatiotemporal requirement for rRNA transcription during mammalian cranial NCC development and corresponding tissue-specific threshold sensitivities to disruptions in rRNA transcription in the pathogenesis of congenital craniofacial disorders.

Connection of social anxiety to impaired pattern of cognitive control and underlying motivational deficiencies: Evidence from event-related potentials.

Numerous studies have established a correlation between social anxiety and poor cognitive control. However, little is known about the cognitive control pattern of individuals with high social anxiety (HSAs) and the underlying mechanisms. Based on the Dual Mechanisms of Control framework and the Expected Value of Control theory, this study explored whether HSAs have an impaired cognitive control pattern (Experiment 1) and whether motivational deficiencies underlie the impaired control pattern (Experiment 2). In Experiment 1, 21 individuals with low social anxiety (LSAs) and 21 HSAs completed an AX-Continuous Performance Task. Results showed that HSAs had a smaller P3b amplitude than LSAs, indicating their weakened proactive control in the cue processing stage, but a larger contingent negative variation (CNV) on cue B as compensation for the negative effects of anxiety in the response preparation stage. No group difference was found in N2 and P3a amplitude on probes, suggesting that reactive control in HSAs was not affected compared to LSAs. In Experiment 2, 21 LSAs and 21 HSAs completed a cued-flanker task, where the likelihood of proactive control engagement was manipulated. The results revealed that HSAs exhibited motivation deficiencies in engaging in proactive control, as evidenced by P3b, CNV amplitude, and response times. These findings shed light on the impaired cognitive control pattern of HSAs and suggest that motivational deficiencies may be the crucial underlying factor.

Mid-arm muscle circumference and triceps skinfold thickness associated with cardiometabolic disease in Chinese residents: A prospective cohort study.

BACKGROUND AND AIMS: The association of cardiometabolic disease (CMD) with body muscle and fat mass remains unclear. Mid-arm muscle circumference (MAMC) and triceps skinfold (TSF) thickness are easily obtained measuring methods for these two body compositions. This study aimed to investigate the association of CMD with MAMC and TSF thickness among Chinese residents. METHODS: A total of 9440 eligible participants from the China Health and Nutrition Survey were included in the analysis. Associations of CMD prevalence with MAMC and TSF thickness were estimated using logistic regression models. Multivariable COX proportional-hazards regression models were used to estimate the effect of baseline MAMC and TSF thickness on subsequent CMD. RESULTS: Positive associations of CMD prevalence with MAMC (odds ratio [OR] = 1.169, 95% confidence interval [CI] 1.110-1.232, P < 0.001) and TSF thickness (OR = 1.313, 95%CI 1.240-1.390, P < 0.001) were observed in the cross-sectional analysis. In the longitudinal study, a 1-SD increase in MAMC was associated with a 13.6% increased risk of CMD incidence (hazard ratio [HR] = 1.136, 95%CI 1.073-1.204, P < 0.001), and a 1-SD increase in TSF thickness had a 17.6% increased risk of CMD incidence (HR = 1.176, 95%CI 1.084-1.276, P < 0.001). For the CMD components, both MAMC and TSF thickness contributed to increased incidences of hypertension (HR = 1.163, 95%CI 1.097-1.233, P < 0.001 in MAMC; HR = 1.218, 95%CI 1.110-1.336, P < 0.001 in TSF thickness) and diabetes mellitus (HR = 1.166, 95%CI 1.028-1.323, P = 0.017 in MAMC; HR = 1.352, 95%CI 1.098-1.664, P = 0.004 in TSF thickness). CONCLUSIONS: Individuals with higher MAMC and TSF thickness had an increased incidence of CMD, mainly hypertension and diabetes mellitus. This study revealed a seemingly counterintuitive association between body muscle mass and metabolic homeostasis. Although the potential mechanisms require further exploration, the impact of body muscle mass on metabolic health cannot be ignored.

Amine-releasable Mediator In situ Repair Perovskites for Efficient and Stable Perovskite Solar Cells.

Residual lead iodide (PbI2) is deemed to a double-edged sword in perovskite film as small amounts of PbI2 are beneficial to the photovoltaic performance, but excessive will cause degradation of photovoltaic performance and stability. Herein, an in situ repair strategy has been developed by introducing amine-releasable mediator (methylammonium pyridine-2-carboxylic, MAPyA) to eliminate over-residual PbI2 and regulate the crystal quality of perovskite film. Notably, MAPyA can be partially decomposed into methylamine (MA) gas and pyridine-2-carboxylic (PyA) during high temperature annealing. The released MA can locally form liquid intermediate phase, facilitating the reconstruction of perovskite microcrystals and residual PbI2. Moreover, the leftover PyA is confirmed to effectively passivate the uncoordinated lead ions in final perovskite film. Based upon this, superior perovskite film with optimized crystal structure and holistic negligible PbI2 is acquired. The assembled device realizes outstanding efficiency of 24.06 %, and exhibits a remarkable operational stability that maintaining 87 % of its origin efficiency after continuous illumination for 1480 h. And the unencapsulated MAPyA-treated devices present significant uplift in humidity stability (maintaining ~93 % of the initial efficiency over 1500 h, 50-60 % relative humidity). Furthermore, the further optimization of this strategy with nanoimprint technology proves its superiority in the amplificative preparation for perovskite films.

gjSOX9 Cloning, Expression, and Comparison with gjSOXs Family Members in Gekko japonicus.

SOX9 plays a crucial role in the male reproductive system, brain, and kidneys. In this study, we firstly analyzed the complete cDNA sequence and expression patterns for SOX9 from Gekko japonicus SOX9 (gjSOX9), carried out bioinformatic analyses of physiochemical properties, structure, and phylogenetic evolution, and compared these with other members of the gjSOX family. The results indicate that gjSOX9 cDNA comprises 1895 bp with a 1482 bp ORF encoding 494aa. gjSOX9 was not only expressed in various adult tissues but also exhibited a special spatiotemporal expression pattern in gonad tissues. gjSOX9 was predicted to be a hydrophilic nucleoprotein with a characteristic HMG-Box harboring a newly identified unique sequence, "YKYQPRRR", only present in SOXE members. Among the 20 SOX9 orthologs, gjSOX9 shares the closest genetic relationships with Eublepharis macularius SOX9, Sphacrodactylus townsendi SOX9, and Hemicordylus capensis SOX9. gjSOX9 and gjSOX10 possessed identical physicochemical properties and subcellular locations and were tightly clustered with gjSOX8 in the SOXE group. Sixteen gjSOX family members were divided into six groups: SOXB, C, D, E, F, and H with gjSOX8, 9, and 10 in SOXE among 150 SOX homologs. Collectively, the available data in this study not only facilitate a deep exploration of the functions and molecular regulation mechanisms of the gjSOX9 and gjSOX families in G. japonicus but also contribute to basic research regarding the origin and evolution of SOX9 homologs or even sex-determination mode in reptiles.

What is the impact of integrated care on the job satisfaction of primary healthcare providers: a systematic review.

BACKGROUND AND OBJECTIVES: The integration of care influenced the job satisfaction of healthcare professionals, especially affecting primary healthcare providers (PCPs). This study aimed to perform a systematic review to explore the impact of integrated care on the job satisfaction of PCPs on the basis of Herzberg's two-factor theory. METHODS: This review was conducted following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We searched 6 electronic databases, including CNKI, WANFANG, PubMed, Web of Science, Cochrane Library, and Embase. Data were retrieved from inception to 19 March 2023. The Mixed Methods Appraisal Tool (MMAT) version 2018 was used to assess the methodological quality of studies for inclusion in the review. RESULTS: A total of 805 articles were retrieved from databases, of which 29 were included in this review. 2 categories, 9 themes, and 14 sub-themes were derived from the data. 2 categories were identified as intrinsic and extrinsic factors. Intrinsic factors included 4 themes: responsibilities, promotion opportunities, recognition, and a sense of personal achievements and growth. Extrinsic factors included 5 themes: salaries and benefits, organizational policy and administration, interpersonal relationships, working conditions, and work status. To specify some key information under certain themes, we also identify sub-themes, such as the sub-theme "workload", "work stress", and "burnout" under the theme "work status". CONCLUSIONS: Findings suggested that the integration of care had both negative and positive effects on the job satisfaction of PCPs and the effects were different depending on the types of integration. Since PCPs played a vital role in the successful integration of care, their job satisfaction was an important issue that should be carefully considered when implementing the integration of care.

Applied phosphorus is maintained in labile and moderately occluded fractions in a typical meadow steppe with the addition of multiple nutrients.

Phosphorus (P) is a limiting nutrient second only to nitrogen (N) in the drylands of the world. Most previous studies have focused on N transformation processes in grassland ecosystems, particularly under artificial fertilization with N and atmospheric N deposition. However, P cycling processes under natural conditions and when P is applied as an inorganic P fertilizer have been understudied. Therefore, it is essential to examine the fate of applied P in grassland ecosystems that have experienced long-term grazing and, under certain circumstances, continuous hay harvest. We conducted a 3-year field experiment with the addition of multiple nutrient elements in a typical meadow steppe to investigate the fate of the applied P in various fractions of P pools in the top soil. We found that the addition of multiple nutrients significantly increased P concentrations in the labile inorganic P (Lab-Pi) and moderately occluded inorganic P (Mod-Pi) fractions but not in the recalcitrant inorganic P (Rec-Pi) fraction. An increase in the concentration of total inorganic P was found only when P and N were applied together. However, the addition of other nutrients did not change P concentrations in any fraction of the mineral soil. The addition of P and N significantly increased the total amount of P taken up by the aboveground plants but had no effect on the levels of organic and microbial P in the soil. Together, our results indicate that the P applied in this grassland ecosystem is taken up by plants, leaving most of the unutilized P as Lab-Pi and Mod-Pi rather than being immobilized in Rec-Pi or by microbial biomass. This implies that the grassland ecosystem that we studied has a relatively low P adsorption capacity, and the application of inorganic P to replenish soil P deficiency in degraded grasslands due to long-term grazing of livestock or continuous harvest of forage in the region could be a practical management strategy to maintain soil P fertility.

Non-cooperative mechanism for bounded and ultrasensitive chromatin remodeling.

Chromatin remodeling is an essential form of gene regulation that is involved in a variety of biological processes. We develop a theoretical model that takes advantage of percolation effects at the level of nucleosome interactions, which allows for ultrasensitive chromatin expansion. This model is non-cooperative and readily provides spatial bounds to the expansion region, preventing uncontrolled remodeling events. We explore different chromatin architectures and the ultrasensitivity of the chromatin density as a function of transcription factor concentration. We also compare our model with experimental data involving an inhibitor of nucleosome acetylation. These results suggest a novel mechanism for spatially-bounded chromatin remodeling and they provide means for quantitative comparisons between proposed models of chromatin architecture.

Enzyme-free and triple-amplified electrochemical sensing of 8-hydroxy-2'-deoxyguanosine by three kinds of short pDNA-driven catalyzed hairpin assemblies followed by a hybridization chain reaction.

A sensitive and enzyme-free electrochemical aptasensor was constructed for the sensing of 8-hydroxy-2'-deoxyguanosine (8-OH-dG). In the process of constructing the aptasensor, triple signal amplification strategies were introduced to enhance the sensitivity. First, every aptamer/pDNA complex immobilized on magnetic beads could release three kinds of pDNAs when 8-OH-dG was introduced, which caused three-fold magnification of the target. Second, the released three kinds of pDNAs initiated catalyzed hairpin assembly between two hairpin DNAs (HP1 and HP2) on a gold electrode. Meanwhile, the three kinds of pDNAs were released again by a strand displacement reaction to obtain the next catalyzed hairpin assembly. Third, the emerging toehold of HP2 further induced a hybridization chain reaction (HCR) between two hairpin DNAs (HP3 and HP4), forming a long double-stranded DNA concatemer on the surface of the electrode. Finally, [Ru(NH3)6]3+, an electroactive cation, was adsorbed onto the long dsDNA concatemer by electrostatic interactions and consequently, an electrochemical signal was generated. Under this triple signal amplification, a low detection limit down to 24.34 fM has been obtained for 8-OH-dG determination, which is superior to those of most previously reported methods.

Cell extrusion - a novel mechanism driving neural crest cell delamination.

Neural crest cells (NCC) comprise a heterogeneous population of cells with variable potency, that contribute to nearly every tissue and organ system throughout the body. Considered unique to vertebrates, NCC are transiently generated within the dorsolateral region of the neural plate or neural tube, during neurulation. Their delamination and migration are crucial events in embryo development as the differentiation of NCC is heavily influenced by their final resting locations. Previous work in avian and aquatic species has shown that NCC delaminate via an epithelial-mesenchymal transition (EMT), which transforms these stem and progenitor cells from static polarized epithelial cells into migratory mesenchymal cells with fluid front and back polarity. However, the cellular and molecular drivers facilitating NCC delamination in mammals are poorly understood. We performed live timelapse imaging of NCC delamination in mouse embryos and discovered a group of cells that exit the neuroepithelium as isolated round cells, which then halt for a short period prior to acquiring the mesenchymal migratory morphology classically associated with most delaminating NCC. High magnification imaging and protein localization analyses of the cytoskeleton, together with measurements of pressure and tension of delaminating NCC and neighboring neuroepithelial cells, revealed these round NCC are extruded from the neuroepithelium prior to completion of EMT. Furthermore, we demonstrate that cranial NCC are extruded through activation of the mechanosensitive ion channel, PIEZO1, a key regulator of the live cell extrusion pathway, revealing a new role for PIEZO1 in neural crest cell development. Our results elucidating the cellular and molecular dynamics orchestrating NCC delamination support a model in which high pressure and tension in the neuroepithelium results in activation of the live cell extrusion pathway and delamination of a subpopulation of NCC in parallel with EMT. This model has broad implications for our understanding of cell delamination in development and disease.

Association between Oral Health and Depressive Symptoms in Chinese Older Adults: The Mediating Role of Dietary Diversity.

Diet is a modifiable factor in healthy population aging. Additionally, oral health and diet are important factors affecting depressive symptoms. To assess the mediating role of dietary diversity (DD) in oral health and depressive symptoms in older adults, we selected 8442 participants aged ≥ 65 years from the 2018 Chinese Longitudinal Health Longevity Survey (CLHLS) for a cross-sectional study. Depressive symptoms were determined based on scores on the 10-item Center for Epidemiologic Studies Depression Scale (CESD-10). Dietary diversity scores (DDS) were established based on the frequency of intake of food groups. Oral health was measured by denture use and toothbrushing frequency. Stepwise multiple linear regression and PROCESS macros were used for mediated effects analysis and testing. The sample had a positive detection rate of 44.1% for depressive symptoms, 40.8% for denture use, and 41.9% for once-a-day toothbrushing. Denture use (ρ = -0.077, p < 0.01) and toothbrushing frequency (ρ = -0.115, p < 0.01) were negative predictors of depressive symptoms in older adults. DD significantly mediated the association between denture use (indirect effect -0.047; 95%CI: -0.068-0.028; p < 0.001), toothbrushing frequency (indirect effect -0.041; 95%CI: -0.054-0.030; p < 0.001), and depressive symptoms. Denture use and toothbrushing frequency not only directly reduce the risk of depressive symptoms in older adults, but also indirectly affect depressive symptoms through DD.

Identification and characterization of intermediate states in mammalian neural crest cell epithelial to mesenchymal transition and delamination.

Epithelial to mesenchymal transition (EMT) is a cellular process that converts epithelial cells to mesenchymal cells with migratory potential in both developmental and pathological processes. Although originally considered a binary event, EMT in cancer progression involves intermediate states between a fully epithelial and a fully mesenchymal phenotype, which are characterized by distinct combinations of epithelial and mesenchymal markers. This phenomenon has been termed epithelial to mesenchymal plasticity (EMP), however, the intermediate states remain poorly described and it's unclear whether they exist during developmental EMT. Neural crest cells (NCC) are an embryonic progenitor cell population that gives rise to numerous cell types and tissues in vertebrates, and their formation is a classic example of developmental EMT. An important feature of NCC development is their delamination from the neuroepithelium via EMT, following which NCC migrate throughout the embryo and undergo differentiation. NCC delamination shares similar changes in cellular state and structure with cancer cell invasion. However, whether intermediate states also exist during NCC EMT and delamination remains unknown. Through single cell RNA sequencing, we identified intermediate NCC states based on their transcriptional signature and then spatially defined their locations in situ in the dorsolateral neuroepithelium. Our results illustrate the progressive transcriptional and spatial transitions from premigratory to migratory cranial NCC during EMT and delamination. Of note gene expression and trajectory analysis indicate that distinct intermediate populations of NCC delaminate in either S phase or G2/M phase of the cell cycle, and the importance of cell cycle regulation in facilitating mammalian cranial NCC delamination was confirmed through cell cycle inhibition studies. Additionally, transcriptional knockdown revealed a functional role for the intermediate stage marker Dlc1 in regulating NCC delamination and migration. Overall, our work identifying and characterizing the intermediate cellular states, processes, and molecular signals that regulate mammalian NCC EMT and delamination furthers our understanding of developmental EMP and may provide new insights into mechanisms regulating pathological EMP.

Core and variable antimicrobial resistance genes in the gut microbiomes of Chinese and European pigs.

Monitoring the prevalence of antimicrobial resistance genes (ARGs) is vital for addressing the global crisis of antibiotic-resistant bacterial infections. Despite its importance, the characterization of ARGs and microbiome structures, as well as the identification of indicators for routine ARG monitoring in pig farms, are still lacking, particularly concerning variations in antimicrobial exposure in different countries or regions. Here, metagenomics and random forest machine learning were used to elucidate the ARG profiles, microbiome structures, and ARG contamination indicators in pig manure under different antimicrobial pressures between China and Europe. Results showed that Chinese pigs exposed to high-level antimicrobials exhibited higher total and plasmid-mediated ARG abundances compared to those in European pigs ( P<0.05). ANT(6)-Ib, APH(3')-IIIa, and tet(40) were identified as shared core ARGs between the two pig populations. Furthermore, the core ARGs identified in pig populations were correlated with those found in human populations within the same geographical regions. Lactobacillus and Prevotella were identified as the dominant genera in the core microbiomes of Chinese and European pigs, respectively. Forty ARG markers and 43 biomarkers were able to differentiate between the Chinese and European pig manure samples with accuracies of 100% and 98.7%, respectively. Indicators for assessing ARG contamination in Chinese and European pigs also achieved high accuracy ( r=0.72-0.88). Escherichia flexneri in both Chinese and European pig populations carried between 21 and 37 ARGs. The results of this study emphasize the importance of global collaboration in reducing antimicrobial resistance risk and provide validated indicators for evaluating the risk of ARG contamination in pig farms.

Identification and characterization of intermediate states in mammalian neural crest cell epithelial to mesenchymal transition and delamination.

Epithelial to mesenchymal transition (EMT) is a cellular process that converts epithelial cells to mesenchymal cells with migratory potential in developmental and pathological processes. Although originally considered a binary event, EMT in cancer progression involves intermediate states between a fully epithelial and a fully mesenchymal phenotype, which are characterized by distinct combinations of epithelial and mesenchymal markers. This phenomenon has been termed epithelial to mesenchymal plasticity (EMP), however, the intermediate states remain poorly described and it's unclear whether they exist during developmental EMT. Neural crest cells (NCC) are an embryonic progenitor cell population that gives rise to numerous cell types and tissues in vertebrates, and their formation and delamination is a classic example of developmental EMT. However, whether intermediate states also exist during NCC EMT and delamination remains unknown. Through single-cell RNA sequencing of mouse embryos, we identified intermediate NCC states based on their transcriptional signature and then spatially defined their locations in situ in the dorsolateral neuroepithelium. Our results illustrate the importance of cell cycle regulation and functional role for the intermediate stage marker Dlc1 in facilitating mammalian cranial NCC delamination and may provide new insights into mechanisms regulating pathological EMP.

Evaluating the link between DIO3-FA27 promoter methylation, biochemical indices, and heart failure progression.

BACKGROUND: Heart failure (HF) is a disease that poses a serious threat to individual health, and DNA methylation is an important mechanism in epigenetics, and its role in the occurrence and development of the disease has attracted more and more attention. The aim of this study was to evaluate the link between iodothyronine deiodinase 3 promoter region fragment FA27 (DIO3-FA27) methylation levels, biochemical indices, and HF. RESULTS: The methylation levels of DIO3-FA27_CpG_11.12 and DIO3-FA27_CpG_23.24 significantly differed in HF patients with different degrees. Multivariate logistic regression analysis indicated that the relative HF risk in the third and fourth quartiles of activated partial thromboplastin time and fibrin degradation products. The results of the restricted cubic spline model showed that the methylation levels of DIO3-FA 27_CpG_11.12 and DIO3-FA 27_CpG_23.24 were associated with coagulation indicators, liver function, renal function, and blood routine. CONCLUSIONS: Based on the differential analysis of CpG methylation levels based on DIO3-FA27, it was found that biochemical indicators combined with DIO3-FA27 promoter DNA methylation levels could increase the risk of worsening the severity classification of HF patients, which provided a solid foundation and new insights for the study of epigenetic regulation mechanisms in patients with HF.

Trait anxiety is related to an impaired attention model for controllable threat cues: Evidence from ERPs.

Attentional bias to threat cues is maladaptive for individuals with high trait anxiety (HTA), but may become adaptive when the dangers signaled by these cues can be controlled by timely actions. However, it remains unclear how HTA individuals allocate attention to controllable threat cues. The current study examined whether trait anxiety is associated with an impaired attention model for controllable threat cues and explored the related underlying neural mechanisms. A sample of 21 participants with low trait anxiety (LTA) and 21 with HTA completed a modified cued anticipation task which allowed participants to control the appearance of threatening pictures associated with controllable threat cues. Results revealed that HTA individuals had no difference in N1 amplitude among controllable threat cues, uncontrollable threat cues, and neutral cues, while LTA individuals showed the greatest N1 amplitude on controllable cues. HTA individuals also exhibited lower N2 amplitude than LTA individuals. The current study provides electrophysiological evidence showing that HTA individuals have impaired attention for processing controllable threat cues and weak inhibitory control. Deficient attention to controllable threat cues may be crucial in the mechanisms underlying trait anxiety.

Targeted Elimination of bla NDM-5 Gene in Escherichia coli by Conjugative CRISPR-Cas9 System.

Purpose: Plasmid-borne carbapenem resistance gene bla NDM-5 accelerates the dissemination of carbapenem-resistant Enterobacteriaceae. To efficiently eliminate the bla NDM-5-harboring plasmid and sensitize the antibiotic-resistant bacteria to meropenem, we used the CRISPR-Cas9 system for combating the carbapenem-resistant Escherichia coli (E. coil). Methods: A series of CRISPR-Cas9 plasmids was constructed, and specific guide RNAs(sgRNA) were designed to target the bla NDM-5 gene. We used chemically transformation or conjugation delivery methods, and the elimination efficiency in each recipient strains was evaluated by plate counting, PCR and quantitative real-time PCR (qPCR). Antimicrobial susceptibility test was carried out by using the broth microdilution method. In addition, we assessed the effect of the CRISPR-Cas9 system of adaptive immunity on the prevention of the exogenous resistant plasmids pNDM-5 by introducing the system into E coli J53. Results: The results showed that pCas9, pCas9-oriT and pBAD-Cas9-oriT can effectively eliminate bla NDM-5 in E. coli with >94.00% elimination efficiency. The bla NDM-5-harboring E. coli successfully restored their susceptibility to meropenem, with eight-fold reduction of minimum inhibitory concentration (MIC) values (from 16 µg/mL to 0.06 µg/mL). The E. coli J53 strain containing plasmid pCas9-N reduced the number of transconjugants by 26-fold. Conclusion: The CRISPR-Cas9 system achieved plasmid clearance and simultaneous re-sensitization to meropenem in E. coli. The CRISPR-Cas9 system could block the horizontal transfer of plasmid pNDM-5. The conjugative delivery of CRISPR-Cas9 provides a new tool for the removal of resistance plasmids and sensitize the recipient to carbapenem. It provides a therapeutic approach to counteract the propagation of bla NDM-5 gene among clinical pathogens.

The Fitting Optimization Path Analysis on Scale Missing Data: Based on the 507 Patients of Poststroke Depression Measured by SDS.

OBJECTIVE: To explore the optimal fitting path of missing data of the Scale to make the fitting data close to the real situation of patients' data. METHODS: Based on the complete data set of the SDS of 507 patients with stroke, the data simulation sets of Missing Completely at Random (MCAR), Missing at Random (MAR), and Missing Not at Random (MNAR) were constructed by R software, respectively, with missing rates of 5%, 10%, 15%, 20%, 25%, 30%, 35%, and 40% under three missing mechanisms. Mean substitution (MS), random forest regression (RFR), and predictive mean matching (PMM) were used to fit the data. Root mean square error (RMSE), the width of 95% confidence intervals (95% CI), and Spearman correlation coefficient (SCC) were used to evaluate the fitting effect and determine the optimal fitting path. RESULTS: when dealing with the problem of missing data in scales, the optimal fitting path is ① under the MCAR deletion mechanism, when the deletion proportion is less than 20%, the MS method is the most convenient; when the missing ratio is greater than 20%, RFR algorithm is the best fitting method. ② Under the Mar mechanism, when the deletion ratio is less than 35%, the MS method is the most convenient. When the deletion ratio is greater than 35%, RFR has a better correlation. ③ Under the mechanism of MNAR, RFR is the best data fitting method, especially when the missing proportion is greater than 30%. In reality, when the deletion ratio is small, the complete case deletion method is the most commonly used, but the RFR algorithm can greatly expand the application scope of samples and save the cost of clinical research when the deletion ratio is less than 30%. The best way to deal with data missing should be based on the missing mechanism and proportion of actual data, and choose the best method between the statistical analysis ability of the research team, the effectiveness of the method, and the understanding of readers.

The co-occurrence of antibiotic resistance genes between dogs and their owners in families.

The intimate relationship between humans and companion animals causes a unique and critical aspect of antimicrobial resistance in humans. However, a comprehensive analysis of antimicrobial resistance between companion animals and their owners is lacking. Here, we chose 13 owned dogs and 16 owners as well as 22 kennel dogs to analyze the effect of an intimate relationship between owned dogs and owners on their gut microbiome, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) and study the correlation of antimicrobial resistance between dogs and their owners in families by metagenomics. Dog gut microbiota had a higher abundance and diversity of ARGs while owners had a higher diversity of taxonomy. In the owned dog gut microbial community, ARG and MGE compositions were significantly more similar to the owner's gut microbiota than those of others. From the perspective of families, there was a strong correlation between macrolide resistance genes between dogs and their owners. In conclusion, our study demonstrated the correlation of ARGs between dogs and their owners at a community-wide level. These findings can alarm the use of antibiotics in companion animals, which implies the potential to harbor antimicrobial resistance and threaten public health.