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Abstract Objective Lung cancer with the highest incidence and mortality in the world. Immune checkpoint inhibitors (ICIs), can bring long-term survival benefits to patients, but also can bring immune-related adverse events (irAEs) in some patients during therapy. Therefore, the aim of this study was to investigate the predictive effect of peripheral blood WBC, NLR, sATPCD4 and nATPCD4 on irAEs in advanced non-small cell lung cancer (NSCLC). Methods Clinical data of 112 patients with advanced NSCLC who were treated with PD -1/PD -L1 inhibitor in the Fifth Affiliated Hospital of Guangzhou Medical University from December 15, 2019 to April 30, 2023 were retrospectively analyzed. These patients were divided into the irAEs group (n = 27) and non-irAEs group (n = 85). The clinical data of the two groups were compared. Receiver operating characteristic (ROC) curves were drawn to determine the threshold value of baseline peripheral blood parameters to predict the occurrence of irAEs. Multivariate logistic regression analysis was used to explore the relationship between peripheral blood markers and the incidence of irAEs. Results The patient characteristics have no significant difference between irAEs and non-irAEs group. But the baseline peripheral blood WBC, sATPCD4 and nATPCD4 of patients in the irAEs group were higher than those in the non-irAEs group (p

Signage is an important medium for individuals to obtain spatial environmental information conveniently and accurately. However, most previous studies focus on signage design for individuals with normal vision, neglecting the specific requirements of those with low vision. Therefore, these signage designs lacking universality restrict the activities of individuals with low vision in public spaces and increase their risks. This study aims to conduct quantitative research on signage size and brightness contrast for low vision individuals. A virtual simulation experimental platform investigated how individuals interpret and react to signage. Two sets of experiments were carried out in Tianjin Key Laboratory of Healthy Habitat and Smart Technology to evaluate the effects of signage size and brightness contrast on response time and accuracy among a total of 139 participants. The results showed that the signage size should be at least 7% of the reading distance to meet the wayfinding needs of low vision individuals. The impact of contrast on wayfinding was strongly dependent on signage size. This research provides valuable insights into the design of signage for low vision individuals.

Abstract Background Many clinical studies based on spontaneous pregnancies (SPs) have demonstrated the superiority of non-invasive prenatal testing (NIPT), and the question of whether this technology is suitable for offspring conceived by assisted reproductive technology has attracted attention. This study aimed to evaluate the application value of NIPT in screening for trisomy (T)21, T18, T13 and sex chromosome aneuploidy (SCA) in pregnant women who conceived by in vitro fertilization (IVF). Results In total, there were 804 high-risk cases [0.88% (804/91280), singleton = 795, twin = 9] in the SP group. Among the 558 invasive prenatal diagnosis (IPD) cases (singleton = 556, twin = 2), 343 (singleton = 342, twin = 1) were true positive, including 213 cases of T21, 28 of T18, 5 of T13 and 97 (singleton = 96, twin = 1) of SCA. The positive predictive values (PPVs) of T21, T18, T13, SCA and T21/T18/T13 combined in singleton pregnancy were 89.12% (213/239), 51.85% (28/54), 21.74% (5/23), 40.00% (96/240), and 77.85% (246/316), respectively, and the PPV of SCA in twin pregnancy was 100.00%. In the IVF group, IPD was performed in 19 (singleton = 16, twin = 3) of the 27 high-risk cases [0.78% (27/3477), singleton = 16, twin = 3], of which 9 (singleton = 8, twin = 1) were true positive, including 5 cases (singleton = 4, twin = 1) of T21 and 4 of SCA. The PPVs of singleton T21, SCA and T21/T18/T13 combined were 66.67% (4/6), 50.00% (4/8) and 57.14% (4/7), respectively, and the PPV of twin T21 was 100.00% (1/1). There were no significant differences in PPV among T21, SCA and T21/T18/T13 combined in singletons between the groups (89.12% vs. 66.67%, p = 0.09; 40.00% vs. 50.00%, p = 0.57; 77.85% vs. 57.14%, p = 0.20). The sensitivity and specificity were higher for singleton and twin pregnancies in the two groups. Based on follow-up results, 1 case of false negative T21 was found in the singleton SP group. Additionally, the mean foetal fraction (FF) of the IVF group was lower than that of the SP group (11.23% vs. 10.51%, p

Background. Sepsis remains a major health condition with a high mortality rate that may be related to immunosuppression. T lymphocyte subsets may reflect the immune function of sepsis patients. The purpose of this study was to investigate the predictive value of CD4+ T lymphocyte counts of ICU patients for their short-term prognosis. Methods. We conducted a prospective, observational cohort study in a general ICU and enrolled patients with sepsis using the Sepsis-3 criteria. Peripheral blood samples were collected within 24 hours of enrollment or measurement of blood cell analysis and biomarkers of CD4+ T lymphocytes and CD8+ T lymphocytes. Severity was classified by the Acute Physiology and Chronic Health Evaluation II and Sequential Organ Failure Assessment (SOFA) scores. The primary outcome was 28-day mortality. Results. A total of 100 patients with sepsis were enrolled and analyzed. CD4+ T lymphocyte counts gradually decreased based on 28-day mortality (p

Abstract BackgroundAlthough evidence-based medicine proposes personalized care that considers the best evidence, it still fails to address personal treatment in many real clinical scenarios where the complexity of the situation makes none of the available evidence applicable. “Medicine-based evidence” (MBE), in which big data and machine learning techniques are embraced to derive treatment responses from appropriately matched patients in real-world clinical practice, was proposed. However, many challenges remain in translating this conceptual framework into practice. ObjectiveThis study aimed to technically translate the MBE conceptual framework into practice and evaluate its performance in providing general decision support services for outcomes after congenital heart disease (CHD) surgery. MethodsData from 4774 CHD surgeries were collected. A total of 66 indicators and all diagnoses were extracted from each echocardiographic report using natural language processing technology. Combined with some basic clinical and surgical information, the distances between each patient were measured by a series of calculation formulas. Inspired by structure-mapping theory, the fusion of distances between different dimensions can be modulated by clinical experts. In addition to supporting direct analogical reasoning, a machine learning model can be constructed based on similar patients to provide personalized prediction. A user-operable patient similarity network (PSN) of CHD called CHDmap ResultsUsing 256 CHD cases, CHDmap was evaluated on 2 different types of postoperative prognostic prediction tasks: a binary classification task to predict postoperative complications and a multiple classification task to predict mechanical ventilation duration. A simple poll of the k ConclusionsWithout individual optimization, CHDmap demonstrates competitive performance compared to clinical experts. In addition, CHDmap has the advantage of enabling clinicians to use their superior cognitive abilities in conjunction with it to make decisions that are sometimes even superior to those made using artificial intelligence models. The MBE approach can be embraced in clinical practice, and its full potential can be realized.

Icons are integral to the signature systems within architectural spaces, serving pivotal roles through human–environment interactions. However, previous icon designs often exhibited a considerable randomness and neglected the needs of visually impaired individuals. To address these issues and to overcome the limitations of the computer-aided design methods, such as most of them focusing only on text design which are not compatible with icons, this study presents an intelligent assistance method named “Universal Colour” for icon colour design based on universal design principles. Such a system enables the rapid generation of icon colour schemes and supports visual and quantitative filtering and comparison during the decision-making process for colour scheme optimization. To assess its usability, fifty-two participants conducted icon colour design experiments using this system, resulting in 87% of the design schemes meeting the universality requirements. The results have demonstrated that Universal Colour has the potential to significantly enhance efficiency and cognitive aspects within the decision-making process for users, regardless of their proficiency in icon design, thereby facilitating the generation of universal icon colour schemes in architectural design.

Abstract Background The tobacco leaf–derived cembratriene-ol exhibits anti-insect effects, but its content in plants is scarce. Cembratriene-ol is difficult and inefficiently chemically synthesised due to its complex structure. Moreover, the titer of reported recombinant hosts producing cembratriene-ol was low and cannot be applied to industrial production. Results In this study, Pantoea ananatis geranylgeranyl diphosphate synthase (CrtE) and Nicotiana tabacum cembratriene-ol synthase (CBTS) were heterologously expressed to synthsize the cembratriene-ol in Escherichia coli. Overexpression of cbts*, the 1-deoxy-d-xylulose 5-phosphate synthase gene dxs, and isopentenyl diphosphate isomerase gene idi promoted the production of cembratriene-ol. The cembratriene-ol titer was 1.53-folds higher than that of E. coli Z17 due to the systematic regulation of ggpps, cbts*, dxs, and idi expression. The production of cembratriene-ol was boosted via the overexpression of genes ispA, ispD, and ispF. The production level of cembratriene-ol in the optimal medium at 72 h was 8.55-folds higher than that before fermentation optimisation. The cembratriene-ol titer in the 15-L fermenter reached 371.2 mg L− 1, which was the highest titer reported. Conclusion In this study, the production of cembratriene-ol in E. coli was significantly enhanced via systematic optimization. It was suggested that the recombinant E. coli producing cembratriene-ol constructed in this study has potential for industrial production and applications.

The reform of education personnel management policies has been carried out progressively to ensure the achievement of educational goals. A systematic analysis of the personnel management policy circle, including policy design, content, and implementation, is crucial to improving the accuracy and effectiveness of such policies. Through a comprehensive review of policy evolution, this study aims to systematize the design and content of policies on personnel management in educational institutions in China. It was determined that personnel management is increasingly concerned with the efficiency and effectiveness of goal achievement, policy design, system reform, and mixed-used management methods. A questionnaire survey was conducted among 420 respondents in 25 preschool institutions in five cities of Henan province in China. A total of 362 questionnaires were deemed valid out of 397 retrieved, including 24 from principals and 338 from teachers. The statistical analysis results identified the problems in policy implementation, including the laxly enforced teacher qualification system, inadequate teacher professional development, poorly implemented training policies, unattractive salaries and benefits, and the undesirable assessment system. For example, 5.89% of the respondents still need teaching qualification certificates. Only 1.1% of the respondents had senior titles. 2.76% of the respondents are still paid less than the minimum standard. In response to the situation, this study suggests effective countermeasures for educational administrative authorities and institutions. The findings could provide a reference for further optimization and development of personnel management policies. AcknowledgmentsThis study is supported by Sumy National Agrarian University in Ukraine and the Talent Special Support Program for Doctoral Studies of Henan Institute of Science and Technology in China.

This paper based on the study of steel plate-fiber reinforced concrete coupling beam components, designs a basic model of composite coupled shear walls with steel plate-fiber reinforced concrete coupling beams. The seismic performance of this model was numerically simulated using the ABAQUS finite element software, analyzing the stress distribution of various parts and the development pattern of plastic hinges in the structure. It also examines how factors such as axial compression ratio, coupling beam cross-sectional dimensions, single-sided wall limb aspect ratio, and total floor height affect the seismic performance of this type of coupled shear wall, providing a reasonable range for axial compression ratios and coupling ratios for composite coupled shear walls with steel plate-fiber reinforced concrete coupling beams. Based on the obtained reasonable axial compression ratio and coupling ratio, and using the analytical solution of the continuum method, a method for controlling the seismic performance of coupled shear wall structures was proposed. Results show that the composite coupled shear walls with steel plate-fiber reinforced concrete coupling beams exhibit high load-bearing capacity and lateral stiffness under inverted triangular horizontal loads, with good displacement ductility and strong energy dissipation capability, making it an excellent seismic performance coupled shear wall system. As the axial compression ratio increases, the displacement corresponding to the yield point and peak point of the structure gradually decreases, and the displacement ductility coefficient shows a trend of first increasing and then decreasing; it is recommended that when designing steel plate-fiber reinforced concrete coupling beam-coupled shear walls, the axial compression ratio should not exceed 0.3. Numerical analysis of coupling ratio-related parameters indicates that in high-intensity seismic design areas, the range of coupling ratios should be between 45% and 60%.

Although there are lots of researches about the application of FRP composites on the external bonding and reinforcement of timber structures, they are mostly aimed at Carbon FRP, which has a large difference in mechanical properties and inconsistent deformation with timber. Meanwhile, research about the interface between timber and Flax Fiber Reinforced Polymer (FFRP) with similar properties with timber is limited; hence, relevant models have not been established yet. Similar to linear elastic FRP confined timber structure, the failure of FFRP-timber structure interface is a process in which the interfacial stress is gradually transferred from the loaded end to the free end, and the interfacial stress transfer area, that is an effective bond length of the FFRP-timber interface. In order to study the interfacial stress transfer characteristics between FFRP and timber structure, based on the existing CFRP-timber structure interface bonding theory and mechanism, a single lap shear test is used to conduct a systematic study on bonding length and width are studied to optimized the application of FFRP. The bond-slip behavior is studied and fitted to provide theory support for the application of FFRP in the restoration of ancient timber structures.

Abstract Background α-Humulene is a plant-derived monocyclic sesquiterpenoid with multiple pharmacological activities, and far-reaching potential for the development of new drugs. Currently, the production of α-humulene is typically achieved via plant extraction, which is not sustainable and limited by low yields. The oleaginous yeast Candida tropicalis has recently emerged as a valuable host for producing high-value-added chemicals. However, the potential of C. tropicalis for terpenoid production has not been exploited. Results In this study, C. tropicalis was engineered for de novo synthesis of α-humulene from glucose. To improve α-humulene production, the codon-optimised α-humulene synthase gene and the entire endogenous farnesyl diphosphate synthesis pathway were co-overexpressed. Furthermore, bottlenecks in the α-humulene synthase pathway were identified and relieved by overexpressing α-humulene synthase, acetoacetyl-CoA thiolase and NADH-dependent HMG-CoA reductase. Combined with fermentation medium optimisation, the engineered strain produced 195.31 mg/L of α-humulene in shake flasks and 4115.42 mg/L in a bioreactor through fed-batch fermentation, a 253- and 5345-fold increase over the initial production, respectively. Conclusions This study demonstrates the potential of C. tropicalis for α-humulene production, and presents a platform for the biosynthesis of other terpenoids.

Abstract Background Gastric cancer (GC), a common malignancy of the human digestive system, represents the second leading cause of cancer-related deaths worldwide. Early detection of GC has a significant impact on clinical outcomes. The aim of this study was to identify potential GC biomarkers. Methods In this study, we conducted a multi-step analysis of expression profiles in GC clinical samples downloaded from TCGA database to identify differentially expressed miRNAs (DEMs) and differentially expressed mRNAs (DEGs). Potential prognostic biomarkers from the available DEMs were then established using the Cox regression method. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to investigate the biological role of the predicted target genes of the miRNA biomarkers. Then, the prognostic DEM-mediated regulatory network was constructed based on transcription factor (TF)–miRNA–target interaction. Subsequently, the consensus genes were further determined based on the overlap between DEGs and these target genes of DEMs. Besides, expression profile, co-expression analysis, immunity, and prognostic values of these prognostic genes were also investigated to further explore the roles in the mechanism of GC tumorigenesis. Results We got five miRNAs, including miR-23b, miR-100, miR-143, miR-145, and miR-409, which are associated with the overall survival of GC patients. Subsequently, enrichment analysis of the target genes of the miRNA biomarkers shown that the GO biological process terms were mainly enriched in mRNA catabolic process, nuclear chromatin, and RNA binding. In addition, the KEGG pathways were significantly enriched in fatty acid metabolism, extracellular matrix (ECM) receptor interaction, and proteoglycans in cancer pathways. The transcriptional regulatory network consisting of 68 TFs, 4 DEMs, and 58 targets was constructed based on the interaction of TFs, miRNAs, and targets. The downstream gene ETS1 of miR-23b and TCF4 regulated by ETS1 were obtained by the regulatory network construction and co-expression analysis. High expression of ETS1 and TCF4 indicated poor prognosis in GC patients, particularly in the advanced stages. The expression of ETS1 and TCF4 was correlated with CD4+ T cells, CD8+ T cells, and B cells. Conclusions miR-23b, ETS1, and TCF4 were identified as the prognostic biomarkers. ETS1 and TCF4 had potential immune function in GC, which provided a theoretical basis for molecular-targeted combined immunotherapy in the future.

Abstract Background Although thousands of long noncoding RNAs (lncRNAs) have been annotated, only a few lncRNAs have been characterized functionally. In this study, we aimed to identify novel lncRNAs involved in the progression of gastric carcinoma (GC) and explore their regulatory mechanisms and clinical significance in GC. Methods A lncRNA expression microarray was used to identify differential lncRNA expression profiles between paired GCs and adjacent normal mucosal tissues. Using the above method, the lncRNA SGO1-AS1 was selected for further study. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and in situ hybridization (ISH) were performed to detect SGO1-AS1 expression in GC tissues. Gain-of-function and loss-of-function analyses were performed to investigate the functions of SGO1-AS1 and its upstream and downstream regulatory mechanisms in vitro and in vivo. Results SGO1-AS1 was downregulated in gastric carcinoma tissues compared to adjacent normal tissues, and its downregulation was positively correlated with advanced clinical stage, metastasis status and poor patient prognosis. The functional experiments revealed that SGO1-AS1 inhibited GC cell invasion and metastasis in vitro and in vivo. Mechanistically, SGO1-AS1 facilitated TGFB1/2 mRNA decay by competitively binding the PTBP1 protein, resulting in reduced TGFβ production and, thus, preventing the epithelial-to-mesenchymal transition (EMT) and metastasis. In addition, in turn, TGFβ inhibited SGO1-AS1 transcription by inducing ZEB1. Thus, SGO1-AS1 and TGFβ form a double-negative feedback loop via ZEB1 to regulate the EMT and metastasis. Conclusions SGO1-AS1 functions as an endogenous inhibitor of the TGFβ pathway and suppresses gastric carcinoma metastasis, indicating a novel potential target for GC treatment.

ABSTRACT Candida tropicalis, a nonmodel diploid microbe, has been applied in industry as a chassis cell. Metabolic engineering of C. tropicalis is challenging due to a lack of gene editing and regulation tools. Here, we report a tRNA:guide RNA (gRNA) platform for boosting gene editing and silencing efficiency in C. tropicalis. As the endogenous tRNA-processing system enables autocleavage for producing a large number of mature gRNAs, a tRNAGly sequence from the genome of C. tropicalis ATCC 20336 was selected for constructing the tRNA:gRNA platform. In the CRISPR-Cas9 system, the tRNA:gRNA platform proved to be efficient in single-gene and multi-gene editing. Furthermore, based on the tRNA:gRNA platform, a CRISPR interference (CRISPRi) system was developed to construct an efficient dCas9-mediated gene expression regulation system for C. tropicalis. The CRISPRi system was employed to regulate the expression of the exogenous gene GFP3 (green fluorescent protein) and the endogenous gene ADE2 (phosphoribosylaminoimidazole carboxylase). Different regions of GFP3 and ADE2 were targeted with the gRNAs processed by the tRNAGly, and the transcription levels of GFP3 and ADE2 were successfully downregulated to 23.9% ± 4.1% and 38.0% ± 7.4%, respectively. The effects of the target regions on gene regulation were also investigated. Additionally, the regulation system was applied to silence ERG9 (squalene synthase) to enhance β-carotene biosynthesis in a metabolically modified C. tropicalis strain. The results suggest that the endogenous tRNAGly and the CRISPRi system have great potential for metabolic engineering of C. tropicalis. IMPORTANCE In the nonmodel yeast Candida tropicalis, a lack of available RNA polymerase type III (Pol III) promoters hindered the development of guide RNA (gRNA) expression platforms for the establishment of CRISPR-Cas-mediated genome editing and silencing strategies. Here, a tRNA:gRNA platform was constructed. We show that this platform allows efficient and precise expression and processing of different gRNAs from a single polycistronic gene capable of mediating multi-gene editing in combination with CRISPR-Cas9. Furthermore, in combination with dCas9, the tRNA:gRNA platform was efficiently used for silencing of exogenous and endogenous genes, representing the first CRISPR interference tool (CRISPRi) in C. tropicalis. Importantly, the established CRISPRi-tRNA:gRNA tool was also used for metabolic engineering by regulating β-carotene biosynthesis in C. tropicalis. The results suggest that the tRNA:gRNA platform and the CRISPRi system will further advance the application of the CRISPR-Cas-based editing and CRISPRi systems for metabolic engineering in C. tropicalis.

Purpose – This study aims to analyze the differences of electrocardiograph (ECG) characteristics for female drivers in calm and anxious states during driving. Design/methodology/approach – The authors used various materials (e.g. visual materials, auditory materials and olfactory materials) to induce drivers’ mood states (calm and anxious), and then conducted the real driving experiments and driving simulations to collect driver’s ECG signal dynamic data. Physiological changes in ECG during the stimulus process were recorded using PSYLAB software. The paired T-test analysis was conducted to determine if there is a significant difference in driver’s ECG characteristics between calm and anxious states during driving. Findings – The results show significant differences in the characteristic parameters of female driver’s ECG signals, including (average heart rate), (atrioventricular interval), (percentage of NN intervals > 50ms), (R wave average peak), (Root mean square of successive), (Q wave average peak) and ( S wave average peak), in time domain, frequency domain and waveform in emotional states of calmness and anxiety. Practical implications – Findings of this work show that ECG can be used to identify driver’s anxious and calm states during driving. It can be used for the development of personalized driver assistance system and driver warning system. Originality/value – Only a few attempts have been made on the influence of human emotions on physiological signals in the transportation field. Hence, there is a need for transport scholars to begin to identify driver’s ECG characteristics under different emotional states. This study will analyze the differences of ECG characteristics for female drivers in calm and anxious states during driving to provide a theoretical basis for developing the intelligent and connected vehicles.

Breast cancer (BRCA) has an undesirable prognosis and is the second most common cancer among women after lung cancer. A novel mechanism of programmed cell death called cuproptosis is linked to the development and spread of tumor cells. However, the function of cuproptosis in BRCA remains unknown. To this date, no studies have used machine learning methods to screen for characteristic genes to explore the role of cuproptosis-related genes (CRGs) in breast cancer. Therefore, 14 cuproptosis-related characteristic genes (CRCGs) were discovered by the feature selection of 39 differentially expressed CRGs using the three machine learning methods LASSO, SVM-RFE, and random forest. Through the PPI network and immune infiltration analysis, we found that PRNP was the key CRCG. The miRTarBase, TargetScan, and miRDB databases were then used to identify hsa-miR-192-5p and hsa-miR-215-5p as the upstream miRNA of PRNP, and the upstream lncRNA, CARMN, was identified by the StarBase database. Thus, the mRNA PRNP/miRNA hsa-miR-192-5p and hsa-miR-215-5p/lncRNA CARMN ceRNA network was constructed. This ceRNA network, which has not been studied before, is extremely innovative. Furthermore, four cuproptosis-related lncRNAs (CRLs) were screened in TCGA-BRCA by univariate Cox, LASSO, and multivariate Cox regression analysis. The risk model was constructed by using these four CRLs, and the risk score = C9orf163 * (1.8365) + PHC2-AS1 * (−2.2985) + AC087741.1 * (−0.9504) + AL109824.1 * (0.6016). The ROC curve and C-index demonstrated the superior predictive capacity of the risk model, and the ROC curve demonstrated that the AUC of 1-, 3-, and 5-year OS in all samples was 0.721, 0.695, and 0.633, respectively. Finally, 50 prospective sensitive medicines were screened with the pRRophetic R package, among which 17-AAG may be a therapeutic agent for high-risk patients, while the other 49 medicines may be suitable for the treatment of low-risk patients. In conclusion, our study constructs a new ceRNA network and a novel risk model, which offer a theoretical foundation for the treatment of BRCA and will aid in improving the prognosis of BRCA.

Nitrile hydratase (NHase, EC 4.2.1.84) is one type of metalloenzyme participating in the biotransformation of nitriles into amides. Given its catalytic specificity in amide production and eco-friendliness, NHase has overwhelmed its chemical counterpart during the past few decades. However, unclear catalytic mechanism, low thermostablity, and narrow substrate specificity limit the further application of NHase. During the past few years, numerous studies on the theoretical and industrial aspects of NHase have advanced the development of this green catalyst. This review critically focuses on NHase research from recent years, including the natural distribution, gene types, posttranslational modifications, expression, proposed catalytic mechanism, biochemical properties, and potential applications of NHase. The developments of NHase described here are not only useful for further application of NHase, but also beneficial for the development of the fields of biocatalysis and biotransformation.

Self-subunit swapping is one of the post-translational maturation of the cobalt-containing nitrile hydratase (Co-NHase) family of enzymes. All of these NHases possess a gene organization of , which allows the activator protein to easily form a mediatory complex with the α-subunit of the NHase after translation. Here, we discovered that the incorporation of cobalt into another type of Co-NHase, with a gene organization of , was also dependent on self-subunit swapping. We successfully isolated a recombinant NHase activator protein (P14K) of Pseudomonas putida NRRL-18668 by adding a Strep-tag N-terminal to the P14K gene. P14K was found to form a complex [α(StrepP14K)(2)] with the α-subunit of the NHase. The incorporation of cobalt into the NHase of P. putida was confirmed to be dependent on the α-subunit substitution between the cobalt-containing α(StrepP14K)(2) and the cobalt-free NHase. Cobalt was inserted into cobalt-free α(StrepP14K)(2) but not into cobalt-free NHase, suggesting that P14K functions not only as a self-subunit swapping chaperone but also as a metallochaperone. In addition, NHase from P. putida was also expressed by a mutant gene that was designed with a order. Our findings expand the general features of self-subunit swapping maturation.

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Clostridioides difficile (C. difficile), as the major pathogen of diarrhea in healthcare settings, has become increasingly prevalent within community populations, resulting in significant morbidity and mortality. However, the therapeutic options for Clostridioides difficile infection (CDI) remain limited, and as of now, no authorized vaccine is available to combat this disease. Therefore, the development of a novel vaccine against C. difficile is of paramount importance. In our study, the complete proteome sequences of 118 strains of C. difficile were downloaded and analyzed. We found four antigenic proteins that were highly conserved and can be used for epitope identification. We designed two vaccines, WLcd1 and WLcd2, that contain the ideal T-cell and B-cell epitopes, adjuvants, and the pan HLA DR-binding epitope (PADRE) sequences. The biophysical and chemical assessments of these vaccine candidates indicated that they were suitable for immunogenic applications. Molecular docking analyses revealed that WLcd1 bonded with higher affinity to Toll-like receptors (TLRs) than WLcd2. Furthermore, molecular dynamics (MD) simulations, performed using Gmx_MMPBSA v1.56, confirmed the binding stability of WLcd1 with TLR2 and TLR4. The preliminary findings suggested that this multi-epitope vaccine could be a promising candidate for protection against CDI; however, experimental studies are necessary to confirm these predictions.

Abstract It is generally believed that Carbapenem-resistant Enterobacterales (CRE) colonization is primarily responsible for systemic infection in humans. However, there is no consensus on whether decolonization should be recommended in clinical practice. In China, the specific situation of CRE colonization and consequent systemic infection in hospitalized patients necessitates further exploration. We conducted a cohort study and analyzed various clinical characteristics of inpatients with intestinal CRE colonization. A risk prediction model for consequent CRE infection was established and externally validated. Our prediction model is freely available online at https://creinfection.shinyapps.io/dynnomapp/ . 839 intestinal CRE colonization samples from inpatients were included. 317 cases of intestinal CRE colonization were enrolled, 25.9% of whom developed systemic infections. The consequent CRE infection rates of Klebsiella pneumoniae and Escherichia coli were 27.0% and 32.3%. The departments at high risk for subsequent CRE infection were respiratory medicine, hematology, and intensive care unit. Secondary infection after intestinal CRE colonization in inpatients can significantly prolong the length of hospital stay (26 days vs. 33 days, P < 0.001), increase the total medical cost (144735.34￥ vs. 281852.34￥, P < 0.001), and has poor (85.11% vs. 52.44%, P < 0.001) efficacy and high mortality (5.96% vs. 18.29%, P = 0.001). Our study makes a significant contribution to comprehensively specify CRE infection, because these results can facilitate early identification of high-risk hospitalized patients, timely implementation to decolonize treatment interventions, ultimately achieve the goal of CRE nosocomial infection prevention and control.

Abstract The aim of this study is to investigate the potential causal relationships between coronary artery disease (CAD), myocardial infarction (MI), urate levels, and aortic aneurysm (AA), abdominal aortic aneurysm (AAA), thoracic aortic aneurysm(TAA), aortic dissection (AD) in individuals of European ancestry. To examine the potential causal relationships between CAD, MI, and urate levels with AA, AAA, TAA, AD, respectively, we performed a two-sample Mendelian randomization (MR) analysis. Genetic instruments that reached genome-wide significance (p

Abstract Background The prevalence of clinically diagnosed depressive disorders (DD) in Chinese left-behind children (LBC) remains unknown. We aim to estimate the prevalence of DD, discuss the associations between DD and self-harm (SH) behaviors in a large representative sample of Chinese LBCs chosen from Yunnan province. Methods A total of 5462 LBCs were selected from the most recent datasets of the Mental Health Survey for Children and Adolescents in Yunnan (MHSCAY), a mega population-based two-phase cross-sectional survey. Weighted prevalence rates and designed Logistic regression were adopted to estimate the prevalence of DD and the association between DD and SH. Results The weighted prevalence of lifetime and current DD were 4.22% (95% CI: 3.13-6.00%) and 3.84% (95% CI: 2.85-5.00%) in Chinese LBCs. Higher lifetime and current DD prevalence rates were observed in girls and those reported adverse parental marital status and SH behaviors. The absence of DD was associated with significantly decreased odds of SH behavior (OR = 0.06), repetitive SH (OR = 0.09), using multiple SH methods (OR = 0.09), and severe SH (OR = 0.15). Subsequently performed stratified analyses identified prominent effect modification by sex and age, as a stronger association between DD and SH was found in girls (OR = 0.02 versus OR = 0.07 in boys) and younger adolescents (OR = 0.08 versus OR = 0.22 in older adolescents). Conclusion The prevalence of DD was high in Chinese LBCs. DD was associated with prominently increased risk of SH behaviors in LBCs. Attention and intervention are needed in this vulnerable population.

Abstract Lung adenocarcinoma (LUAD), characterized by its heterogeneity and complex pathogenesis, is the focus of this study which investigates the association between cell death-related genes and LUAD. Through machine learning, a risk score model was developed using the Coxboost rsf algorithm, demonstrating strong prognostic accuracy in both validation (GSE30219, GSE31210, GSE72094) and training (TCGA-LUAD) datasets with C-indices of 0.93, 0.67, 0.68, and 0.64, respectively. The study reveals that the expression of Keratin 18 (KRT18), a key cytoskeletal protein, varies across LUAD cell lines (DV-90, PC-9, A549) compared to normal bronchial epithelial cells (BEAS-2B), suggesting its potential role in LUAD's pathogenesis. Kaplan–Meier survival curves further validate the model, indicating longer survival in the low-risk group. A comprehensive analysis of gene expression, functional differences, immune infiltration, and mutations underscores significant variations between risk groups, highlighting the high-risk group's immunological dysfunction. This points to a more intricate tumor immune environment and the possibility of alternative therapeutic strategies. The study also delves into drug sensitivity, showing distinct responses between risk groups, underscoring the importance of risk stratification in treatment decisions for LUAD patients. Additionally, it explores KRT18's epigenetic regulation and its correlation with immune cell infiltration and immune regulatory molecules, suggesting KRT18's significant role in the tumor immune landscape. This research not only offers a valuable prognostic tool for LUAD but also illuminates the complex interplay between cell death-related genes, drug sensitivity, and immune infiltration, positioning KRT18 as a potential therapeutic or prognostic target to improve patient outcomes by personalizing LUAD treatment strategies.

Abstract Background Existing evidence suggests that children and adolescents who had experienced childhood maltreatment (CM) are at higher suicidal risk. However, the mediation role of resilience in this association remains unclear. We aim to investigate the mediation via resilience in the associations between CM and three suicidal risk indicators (suicidal ideation, SI; suicidal plan, SP; suicidal attempt, SA) among a large sample of Chinese children and adolescents. Methods A population-based cross-sectional survey was conducted in southwestern China Yunnan province. A total of 9723 children and adolescents were included and analyzed by using a multi-stage stratified cluster sampling design. Univariate and multivariate logistic regression models were fitted to explore the associations between CM, resilience, and the three suicidal risk indicators, dose-response trends further elucidated by using the restricted cubic splines. Path models were adopted to estimate the mediation of resilience. Results The estimated prevalence rates for one-year SI, SP and SA were 32.86% (95% CI: 31.93–33.80%), 19.36% (95% CI: 18.57–20.16%) and 9.07% (95% CI: 8.51–9.66%). After adjustment, CM significantly associated with all 3 suicidal risk indicators, and the odds ratios (ORs) were 2.13 (95% CI: 1.91–2.37), 2.45 (95% CI: 2.13–2.81), and 3.61 (95% CI: 2.90–4.52) for one-year SI, SP, and SA, respectively. Path models revealed that resilience significantly mediated the associations between CM and the three suicidal risk indicators, and among all dimensions of resilience, family support presented the strongest mediation consistently. Conclusions Our study results suggest that intervention measures which focusing on improving psychological resilience might be effective in reducing suicidal risk for children and adolescents who had experienced maltreatment. Prospective studies should be done to corroborate our findings.

Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection. Breeding of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this disease. The traditional way of R gene functional validation requires stable transformation that is both time- and labor-consuming. In this study, a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was developed. The transformation positive rate was over 80% in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable transformation. The system was applicable to different B. napus varieties and other cruciferous crops including Brassica rapa and Brassica oleracea. In particular, two known CR genes, CRA3.7.1 and CRA8.2.4 were used respectively, as example to show that the system works well for CR gene study combined with subsequent P. brassicae infection in B. napus. Most importantly, it works both in over-expression that led to disease resistance, as well as in RNAi which led to disease susceptible phenotype. Therefore, this system can be used in batch-wise identification of CR genes, and also offered the possibility of manipulating key genes within the P. brassicae genome that could improve our knowledge on host–pathogen interaction.

Abstract Background The advent of immunotherapy targeting immune checkpoints has conferred significant clinical advantages to patients with lung adenocarcinoma (LUAD); However, only a limited subset of patients exhibit responsiveness to this treatment. Consequently, there is an imperative need to stratify LUAD patients based on their response to immunotherapy and enhance the therapeutic efficacy of these treatments. Methods The differentially co-expressed genes associated with CD8 + T cells were identified through weighted gene co-expression network analysis (WGCNA) and the Search Tool for the Retrieval of Interacting Genes (STRING) database. These gene signatures facilitated consensus clustering for TCGA-LUAD and GEO cohorts, categorizing them into distinct immune subtypes (C1, C2, C3, and C4). The Tumor Immune Dysfunction and Exclusion (TIDE) model and Immunophenoscore (IPS) analysis were employed to assess the immunotherapy response of these subtypes. Additionally, the impact of inhibitors targeting five hub genes on the interaction between CD8 + T cells and LUAD cells was evaluated using CCK8 and EDU assays. To ascertain the effects of these inhibitors on immune checkpoint genes and the cytotoxicity mediated by CD8 + T cells, flow cytometry, qPCR, and ELISA methods were utilized. Results Among the identified immune subtypes, subtypes C1 and C3 were characterized by an abundance of immune components and enhanced immunogenicity. Notably, both C1 and C3 exhibited higher T cell dysfunction scores and elevated expression of immune checkpoint genes. Multi-cohort analysis of Lung Adenocarcinoma (LUAD) suggested that these subtypes might elicit superior responses to immunotherapy and chemotherapy. In vitro experiments involved co-culturing LUAD cells with CD8 + T cells and implementing the inhibition of five pivotal genes to assess their function. The inhibition of these genes mitigated the immunosuppression on CD8 + T cells, reduced the levels of PD1 and PD-L1, and promoted the secretion of IFN-γ and IL-2. Conclusions Collectively, this study delineated LUAD into four distinct subtypes and identified five hub genes correlated with CD8 + T cell activity. It lays the groundwork for refining personalized therapy and immunotherapy strategies for patients with LUAD.

Abstract Background Aberrant activation of macrophages is associated with pathogenesis of acute lung injury (ALI). However, the potential pathogenesis has not been explored. Objectives We aimed to identify whether histone deacetylase (HDAC) 10 is involved in lipopolysaccharide (LPS)-exposed ALI and reveal the underlying pathogenesis by which it promotes lung inflammation in LPS-exposed ALI via modifying P62 with deacetylation. Methods We constructed an ALI mice model stimulated with LPS to determine the positive effect of Hdac10 deficiency. Moreover, we cultured murine alveolar macrophage cell line (MH-S cells) and primary bone marrow-derived macrophages (BMDMs) to explore the pro-inflammatory activity and mechanism of HDAC10 after LPS challenge. Results HDAC10 expression was increased both in mice lung tissues and macrophage cell lines and promoted inflammatory cytokines production exposed to LPS. Hdac10 deficiency inhibited autophagy and inflammatory response after LPS stimulation. In vivo, Hdac10 fl/fl -LysMCre mice considerably attenuated lung inflammation and inflammatory cytokines release exposed to LPS. Mechanistically, HDAC10 interacts with P62 and mediates P62 deacetylation at lysine 165 (K165), by which it promotes P62 expression and increases inflammatory cytokines production. Importantly, we identified that Salvianolic acid B (SAB), an HDAC10 inhibitor, reduces lung inflammatory response in LPS-stimulated ALI. Conclusion These results uncover a previously unknown role for HDAC10 in regulating P62 deacetylation and aggravating lung inflammation in LPS-induced ALI, implicating that targeting HDAC10 is an effective therapy for LPS-exposed ALI.

ObjectiveLung cancer is the leading cause of cancer-related mortality. Lymph node involvement remains a crucial prognostic factor in non-small cell lung cancer (NSCLC), and the TNM system is the current standard for staging. However, it mainly considers the anatomical location of lymph nodes, neglecting the significance of node count. Metrics like metastatic lymph node count and lymph node ratio (LNR) have been proposed as more accurate predictors.MethodsWe used data from the SEER 17 Registry Database (2010–2019), including 52,790 NSCLC patients who underwent lobectomy or pneumonectomy, with at least one lymph node examined. Primary outcomes were overall survival (OS) and cancer-specific survival (CSS). Cox regression models assessed the prognostic value of negative lymph node (NLN) count, number of positive lymph node (NPLN), and LNR, with cut-points determined using X-tile software. Model performance was evaluated by the Akaike information criterion (AIC).ResultsThe Cox proportional hazards model analysis revealed that NLN, NPLN, and LNR are independent prognostic factors for OS and LCSS (P

Background Serum neuron-specific enolase (NSE) had been associated with survival of several cancers. However, its prognostic significance for colorectal cancer (CRC) has not been effectively discussed. We aimed to investigate the relationship between baseline serum NSE and the overall survival (OS) of colorectal adenocarcinoma (CRAD) patients. Methods A retrospective study had been conducted by including 564 histopathology confirmed CRAD patients between January 2013 and December 2018 from Yunnan Provincial Cancer hospital, China. Cox proportional hazards model was used to estimate the crude and adjusted associations between serum NSE measured at diagnosis and the OS of the patients. Restricted cubic spline (RCS) was further applied to delineate dose-response trend of the NSE-OS association. Results After controlling for possible confounding factors, baseline serum NSE was significantly associated with OS in CRAD: when dichotomizing by the median, patients with higher baseline serum NSE (NSE >= 12.93 ng/mL) were observed a worse prognosis (hazard ratio, HR: 1.82, 95% CI [1.30–2.55], p < 0.01). Stratified analysis by tumor stage revealed a stronger NSE-OS association in advanced CRAD patients. RCS disclosed a prominent dose-response relationship in NSE-OS association for all CRAD patients: along with the increase of baseline serum NSE, the adjusted HR of CRAD patients increased gradually. This dose-response trend is also evident in advanced stage CRAD patients, but not in early stage CRAD patients. Conclusions Serum NSE measured at diagnosis might be a useful prognostic indicator for CRAD, especially for advanced stage patients.

BackgroundCantharidin (CTD) extracted from the traditional Chinese medicine Mylabris has significant therapeutic effects on various tumors. However, the high toxicity of CTD can cause serious liver damage, although the related molecular mechanisms remain unclear.MethodsIn this study, we established models of CTD-induced liver and L-O2 cell damage in mice in vivo and in vitro. Subsequently, liver function indicators were detected in mouse serum, while liver tissues were subjected to pathological and transmission electron microscopy observations. L-O2 cell activity was investigated using the CCK-8 assay, and the mRNA and protein expression of DNA damage-induced transcription factor 4 (DDIT4) in liver tissue and L-O2 cells was detected using qPCR, immunohistochemistry, and western blotting. Western blotting was also used to detect the expression levels of autophagy- and apoptosis-related proteins in liver tissue and L-O2 cells. After RNAi interference with DDIT4, Rap, and 3-MA treatment, autophagy and apoptosis of L-O2 cells were detected using western blotting, flow cytometry, transmission electron microscopy, and confocal microscopy.ResultsFollowing CTD exposure, the mouse liver showed significant pathological damage and an increase in autophagic lysosomes, while the vitality of L-O2 cells showed a significant decrease. CTD led to a significant increase in the mRNA and protein levels of DDIT4 in both liver tissue and L-O2 cells, as well as a significant increase in LC3-II, Beclin1, and Bax, whereas p-mTOR and Bcl-2 were significantly decreased. Following DDIT4 interference and 3-MA treatment, the levels of autophagy and apoptosis induced by CTD in L-O2 cells were reduced. After Rap treatment, both autophagy and apoptosis of CTD-induced L-O2 cells were significantly enhanced.ConclusionThe molecular mechanism of CTD-induced toxicity in mouse liver and L-O2 cells is mainly through DDIT4/mTOR signaling pathway activation, leading to an increase in autophagy and apoptosis levels.

Objective In this study, we investigated the cause of the AngII dose elevation in aqueous humor of primary open-angle glaucoma (POAG) patients. Methods Enzyme-linked immunosorbent assay (ELISA), western blotting were used to detect concentration of Angiotensin Converting Enzyme 2 (ACE2) and Prolylcarboxypeptidase (PRCP). AngII and AngII + Recombinant PRCP were injected into anterior chamber of mouse eye. Mouse Intraocular pressure (IOP) was measured every week, mouse eye sections were conducted Hematoxylin-and-Eosin (H&E) staining, Masson’ staining and Immunofluorescence staining. Western blotting and Immunofluorescence staining assays to detected fibrosis of trabecular meshwork cells. Mass spectrometry was used to identify proteins of aqueous humor. Results PRCP dose are decreased in aqueous humor of POAG patients. There is a negative correlation between PRCP and AngII levels in aqueous humor and between PRCP levels and the IOP. PRCP treatment reverses fibrosis of trabecular meshwork (TM) and prevents IOP elevation induced by AngII. Exogenous PRCP rescues fibrosis induced by AngII in HTMCs. Proteome profiling detected 502 differentially expressed proteins. Conclusion Our study found PRCP dose was decreased in POAG patients’ aqueous humor, and it might cause high level of AngII. Restoration of PRCP rescued fibrosis of TM cells and ameliorated IOP in AngII treatment mouse.