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Abstract Background Early dissemination to distant organs accounts for the dismal prognosis of patients with pancreatic ductal adenocarcinoma (PDAC). Chronic, dysregulated, persistent and unresolved inflammation provides a preferred tumor microenvironment (TME) for tumorigenesis, development, and metastasis. A better understanding of the key regulators that maintain inflammatory TME and the development of predictive biomarkers to identify patients who are most likely to benefit from specific inflammatory-targeted therapies is crucial for advancing personalized cancer treatment. Methods This study identified cell-specific expression of CALB2 in human PDAC through single-cell RNA sequencing analysis and assessed its clinicopathological correlations in tissue microarray using multi-color immunofluorescence. Co-culture systems containing cancer-associated fibroblasts (CAFs) and patient-derived organoids (PDOs) in vitro and in vivo were employed to elucidate the effects of CALB2-activated CAFs on PDAC malignancy. Furthermore, CUT&RUN assays, luciferase reporter assays, RNA sequencing, and gain- or loss-of-function assays were used to unravel the molecular mechanisms of CALB2-mediated inflammatory reprogramming and metastasis. Additionally, immunocompetent KPC organoid allograft models were constructed to evaluate CALB2-induced immunosuppression and PDAC metastasis, as well as the efficacy of inflammation-targeted therapy. Results CALB2 was highly expressed both in CAFs and cancer cells and correlated with an unfavorable prognosis and immunosuppressive TME in PDAC patients. CALB2 collaborated with hypoxia to activate an inflammatory fibroblast phenotype, which promoted PDAC cell migration and PDO growth in vitro and in vivo. In turn, CALB2-activated CAFs upregulated CALB2 expression in cancer cells through IL6-STAT3 signaling-mediated direct transcription. In cancer cells, CALB2 further activated Ca2+-CXCL14 inflammatory axis to facilitate PDAC metastatic outgrowth and immunosuppression. Genetic or pharmaceutical inhibition of CXCL14 significantly suppressed CALB2-mediated metastatic colonization of PDAC cells in vivo and extended mouse survival. Conclusions These findings identify CALB2 as a key regulator of inflammatory reprogramming to promote PDAC metastatic progression. Combination therapy with αCXCL14 monoclonal antibody and gemcitabine emerges as a promising strategy to suppress distant metastasis and improve survival outcomes in PDAC with CALB2 overexpression.

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy with a poor prognosis and limited therapeutic options. Research on the tumor microenvironment (TME) of PDAC has propelled the development of immunotherapeutic and targeted therapeutic strategies with a promising future. The emergence of single-cell sequencing and mass spectrometry technologies, coupled with spatial omics, has collectively revealed the heterogeneity of the TME from a multiomics perspective, outlined the development trajectories of cell lineages, and revealed important functions of previously underrated myeloid cells and tumor stroma cells. Concurrently, these findings necessitated more refined annotations of biological functions at the cell cluster or single-cell level. Precise identification of all cell clusters is urgently needed to determine whether they have been investigated adequately and to identify target cell clusters with antitumor potential, design compatible treatment strategies, and determine treatment resistance. Here, we summarize recent research on the PDAC TME at the single-cell multiomics level, with an unbiased focus on the functions and potential classification bases of every cellular component within the TME, and look forward to the prospects of integrating single-cell multiomics data and retrospectively reusing bulk sequencing data, hoping to provide new insights into the PDAC TME.

Abstract Background Haematological patients exhibit immune system abnormalities that make them susceptible to viral infections. Understanding the relationship between the virome in the blood plasma of haematological patients and their clinical characteristic is crucial for disease management. We aimed to explore the presence of viral pathogens and identify close associations between viral infections and various clinical features. Results A total of 21 DNA viruses and 6 RNA viruses from 12 virus families were identified from 1383 patients. Patients with haematological diseases exhibited significantly higher diversity, prevalence, and co-detection rates of viral pathogens. During fever episodes, pathogen detection was notably higher, with Epstein-Barr virus (EBV) and Mucorales infections being the most probable culprits for fever symptoms in non-haematological patients. The detection rate of torque teno virus (TTV) significantly increases in haematological patients after transplantation and during primary lung infections. Additionally, TTV-positive patients demonstrate significantly higher absolute neutrophil counts, while C-reactive protein and procalcitonin levels are notably lower. Furthermore, TTV, cytomegalovirus, and parvovirus B19 (B19V) were found to be more prevalent in non-neutropenic patients, while non-viral pathogenic infections, such as Gram-negative bacteria and Mucorales, were more common in neutropenic patients. Pegivirus C (HPgV-C) infection often occurred post-transplantation, regardless of neutropenia. Additionally, some viruses such as TTV, B19V, EBV, and HPgV-C showed preferences for age and seasonal infections. Conclusions Analysis of the plasma virome revealed the susceptibility of haematological patients to plasma viral infections at specific disease stages, along with the occurrence of mixed infections with non-viral pathogens. Close associations were observed between the plasma virome and various clinical characteristics, as well as clinical detection parameters. Understanding plasma virome aids in auxiliary clinical diagnosis and treatment, enabling early prevention to reduce infection rates in patients and improve their quality of life. Video Abstract

Objective: To explore the association between retinal microvascular parameters and glaucoma. Design: Prospective study. Subjects: The UK Biobank subjects with fundus images and without a history of glaucoma. Methods: We employed the Retina-based Microvascular Health Assessment System to utilize the noninvasive nature of fundus photography and quantify retinal microvascular parameters including retinal vascular skeleton density (VSD) and fractal dimension (FD). We also utilized propensity score matching (PSM) to pair individuals with glaucoma and healthy controls. Propensity score matching was implemented via a logistic regression model with a caliper of 0.1 and a matching ratio of 1:4 no replacements. We conducted univariable Cox regression analyses to study the association between retinal microvascular parameters and incident glaucoma, in both continuous and quartile forms. Main Outcome Measure: Vascular skeleton density, FD, and glaucoma. Results: In a study of 41 632 participants without prior glaucoma, 482 cases of glaucoma were recorded during a median follow-up of 11.0 years. In the Cox proportional hazards regression model post-PSM, we found that incident glaucoma has significant negative associations with arteriolar VSD (hazard ratio [HR] = 0.24, 95% confidence interval [CI] 0.11–0.52, P

Summary: Background: The global spread of the plasmid-mediated mcr (mobilized colistin resistance) gene family presents a significant threat to the efficacy of colistin, a last-line defense against numerous Gram-negative pathogens. The mcr-9 is the second most prevalent variant after mcr-1. Methods: A dataset of 698 mcr-9-positive isolates from 44 countries is compiled. The historical trajectory of the mcr-9 gene is reconstructed using Bayesian analysis. The effective reproduction number is used innovatively to study the transmission dynamics of this mobile-drug-resistant gene. Findings: Our investigation traces the origins of mcr-9 back to the 1960s, revealing a subsequent expansion from Western Europe to the America and East Asia in the late 20th century. Currently, its transmissibility remains high in Western Europe. Intriguingly, mcr-9 likely emerged from human-associated Salmonella and exhibits a unique propensity for transmission within the Enterobacter. Our research provides a new perspective that this host preference may be driven by codon usage biases in plasmids. Specifically, mcr-9-carrying plasmids prefer the nucleotide C over T compared to mcr-1-carrying plasmids among synonymous codons. The same bias is seen in Enterobacter compared to Escherichia (respectively as their most dominant genus). Furthermore, we uncovered fascinating patterns of coexistence between different mcr-9 subtypes and other resistance genes. Characterized by its low colistin resistance, mcr-9 has used this seemingly benign feature to silently circumnavigate the globe, evading conventional detection methods. However, colistin-resistant Enterobacter strains with high mcr-9 expression have emerged clinically, implying a strong risk of mcr-9 evolving into a global “true-resistance-gene”. Interpretation: This study explores the mcr-9 gene, emphasizing its origin, adaptability, and dissemination potential. Given the high mcr-9 expression colistin-resistant strains was observed in clinically the prevalence of mcr-9 poses a significant challenge to drug resistance prevention and control within the One Health framework. Funding: This work was partially supported by the National Natural Science Foundation of China (Grant No. 32141001 and 81991533).

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a highly malignant neoplasm characterized by a poor prognosis and limited therapeutic strategy. The PDAC tumor microenvironment presents a complex heterogeneity, where neutrophils emerge as the predominant constituents of the innate immune cell population. Leveraging the power of single-cell RNA-seq, spatial RNA-seq, and multi-omics approaches, we included both published datasets and our in-house patient cohorts, elucidating the inherent heterogeneity in the formation of neutrophil extracellular traps (NETs) and revealed the correlation between NETs and immune suppression. Meanwhile, we constructed a multi-omics prognostic model that suggested the patients exhibiting downregulated expression of NETs may have an unfavorable outcome. We also confirmed TLR2 as a potent prognosis factor and patients with low TLR2 expression had more effective T cells and an overall survival extension for 6 months. Targeting TLR2 might be a promising strategy to reverse immunosuppression and control tumor progression for an improved prognosis.

Objectives: Although anti-VEGF and retinal laser photocoagulation are two therapeutic modalities that have been used in the clinical treatment of diabetic retinopathy (DR), it is unknown how these modalities target vascular endothelial function in DR. Methods: We first downloaded and analyzed the differential genes in two DR-related datasets, GSE60436 and GSE53257. The differential gene expression was then verified using RT-qPCR, and the most upregulated gene, NDUFB7, was selected for subsequent experiments. Subsequently, the role of NDUFB7 silencing and enforced expression on the proliferation and apoptosis of HRVECs was explored using CCK-8 assay, EDU proliferation assay and apoptotic TUNEL staining. In addition, the upstream potential miRNAs of NDUFB7 were predicted online using the Targetscan website. RT-qPCR, Western blotting (WB), and dual luciferase gene reporter assay were used to confirm the targeting connection between miR-2861 and NDUFB7. Finally, miR-2861 expression after high glucose (HG) treatment and its effect on proliferation and apoptosis of HRVECs under HG were investigated. Results: In this study, we first downloaded and analyzed the differential genes in two DR-related datasets, GSE60436 and GSE53257. We found that TUFM, PRELID1, MRPL32, NDUFB7, MRPL4, MRPL40, HSD17B10 and SLC25A13 were upregulated in DR, and RT-qPCR showed that NDUFB7 was most upregulated. Subsequent CCK-8 assay, EDU proliferation assay and TUNEL staining showed that up-picked NDUFB7 promotes proliferation and inhibits apoptosis of HRVECs. In addition, the upstream potential miRNAs of NDUFB7 were predicted online using the Targetscan website. RT-qPCR, Western blotting (WB), and dual luciferase gene reporter assay confirmed the targeting connection between miR-2861 and NDUFB7. Finally, it was observed that miR-2861 can inhibit the proliferation and promote the apoptosis of HRVECs by targeting NDUFB7. Conclusions: Our findings showed that upregulated NDUFB7 in DR promotes proliferation and inhibits apoptosis of HRVECs, and miR-2861 can rescue the pathogenic effect of NDUFB7 upregulation by targeting NDUFB7.

BackgroundDepression and anxiety are common mental health problems among adolescents worldwide. Extant research has found that intelligence, emotion, and failure-is-debilitating beliefs (fixed mindsets) are closely related to more depression and anxiety symptoms, hopelessness, and suicidality. Recent research also points to the importance of parental mindset, which can strongly influence children’s affect, behavior, and mental health. However, the effects of parent-child mindset interventions on a child’s internalizing problems have not yet been empirically examined. As recent evidence has shown the promise of single-session interventions in reducing and preventing youth internalizing problems, this study develops and examines a parent and child single-session intervention on mindsets of intelligence, failure, and emotion (PC-SMILE) to tackle depression and anxiety in young people. ObjectiveUsing a 3-arm randomized controlled trial, this study will examine the effectiveness of PC-SMILE in reducing depression and anxiety symptoms among children. We hypothesize that compared to the waitlist control group, the PC-SMILE group and child single-session intervention on mindsets of intelligence, failure, and emotion (C-SMILE) group will significantly improve child depression and anxiety (primary outcome) and significantly improve secondary outcomes, including children’s academic self-efficacy, hopelessness, psychological well-being, and parent-child interactions and relationships, and the PC-SMILE is more effective than the C-SMILE. MethodsA total of 549 parent-child dyads will be recruited from 8 secondary schools and randomly assigned to either the PC-SMILE intervention group, the C-SMILE intervention group, or the no-intervention waitlist control group. The 45-minute interventions include parent-version and child-version. Both parents and students in the PC-SMILE group receive the intervention. Students in C-SMILE group receive intervention and their parents will receive intervention after all follow-up ends. Students in 3 groups will be assessed at 3 time points, baseline before intervention, 2 weeks post intervention, and 3 months post intervention, and parents will be assessed in baseline and 3-month follow-up. The intention-to-treat principle and linear-regression-based maximum likelihood multilevel models will be used for data analysis. ResultsRecruitment started in September 2023. The first cohort of data collection is expected to begin in May 2024 and the second cohort will begin in September 2024. The final wave of data is expected to be collected by the end of the first quarter of 2025. The results are expected to demonstrate improved anxiety and depression among students assigned to the intervention condition, as well as the secondary outcomes compared to those in the control group. The efficacy and effectiveness of the intervention will be discussed. ConclusionsThis study is the first attempt to develop a web-based single-session intervention for students and their parents to enhance their well-being in Hong Kong and beyond, which potentially contributes to providing evidence-based recommendations for the implementation of brief digital parent-child interventions. International Registered Report Identifier (IRRID)PRR1-10.2196/63220

Summary: Klebsiella pneumoniae carbapenemase (KPC) poses a major public health risk. Understanding its transmission dynamics requires examining the epidemiological features of related plasmids. Our study compiled 15,660 blaKPC-positive isolates globally over the past two decades. We found extensive diversity in the genetic background of KPC, with 23 Tn4401-related and 341 non-Tn4401 variants across 163 plasmid types in 14 genera. Intra-K. pneumoniae and cross-genus KPC transmission patterns varied across four distinct periods. In the initial periods, plasmids with narrow host ranges gradually established a survival advantage. In later periods, broad-host-range plasmids became crucial for cross-genera transmission. In total, 61 intra-K. pneumoniae and 66 cross-genus transmission units have been detected. Furthermore, phylogenetic reconstruction dated the origin of KPC transmission back to 1991 and revealed frequent exchanges across countries. Our research highlights the frequent and transient spread events of KPC mediated by plasmids across multiple genera and offers theoretical support for high-risk plasmid monitoring.

Abstract The acquisition of exogenous mobile genetic material imposes an adaptive burden on bacteria, whereas the adaptational evolution of virulence plasmids upon entry into carbapenem-resistant Klebsiella pneumoniae (CRKP) and its impact remains unclear. To better understand the virulence in CRKP, we characterize virulence plasmids utilizing a large genomic data containing 1219 K. pneumoniae from our long-term surveillance and publicly accessible databases. Phylogenetic evaluation unveils associations between distinct virulence plasmids and serotypes. The sub-lineage ST11-KL64 CRKP acquires a pK2044-like virulence plasmid from ST23-KL1 hypervirulent K. pneumoniae, with a 2698 bp region deletion in all ST11-KL64. The deletion is observed to regulate methionine metabolism, enhance antioxidant capacity, and further improve survival of hypervirulent CRKP in macrophages. The pK2044-like virulence plasmid discards certain sequences to enhance survival of ST11-KL64, thereby conferring an evolutionary advantage. This work contributes to multifaceted understanding of virulence and provides insight into potential causes behind low fitness costs observed in bacteria.

Abstract Background Diabetic kidney disease (DKD) is a common and severe complication of diabetes that can lead to end-stage renal disease with no cure. The first-line drugs recommended by clinical guidelines fail to achieve satisfactory effects for people with DKD. A Chinese herbal medicine Tangshen Qushi Formula (TQF) shows preliminary efficacy and safety in preserving renal function for people with DKD, but the effects on comprehensive renal outcomes remain unclear. We will conduct a systematic review and meta-analysis to evaluate the effects of TQF herbs and their compounds identified from ultra-high performance liquid chromatography-MS/MS in diabetic animal models with renal outcomes. Methods This protocol complies with the guideline Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols. We will include studies investigating the effects of TQF herbs and compounds on diabetic rats or mice with renal outcomes. Six electronic databases will be searched from their inception to February 2023. Quality assessment will be conducted using SYRCLE’s risk of bias tool. Standardized or weighted mean differences will be estimated for renal outcomes (creatinine, urea, proteinuria, histological changes, oxidative stress, inflammation, and kidney fibrosis). Data will be pooled using random-effects models. Heterogeneity across studies will be expressed as I 2. Sensitivity analyses will explore treatment effects in adjusted models and within subgroups. Funnel plots and Egger’s test will be used to explore publication bias. Discussion The results of this review will provide valuable insights into the potential effects of TQF in managing DKD. The limitation is that the included studies will be animal studies from specific databases, and the interpretation of the findings must be cautious. Systematic review registration PROSPERO CRD42023432895. Registered on 19 July 2023 ( https://www.crd.york.ac.uk/PROSPERO/#recordDetails ).

Abstract Neoplastic cells need to adapt their gene expression pattern to survive in an ever-changing or unfavorable tumor microenvironment. Protein synthesis (or mRNA translation), an essential part of gene expression, is dysregulated in cancer. The emergence of distinct translatomic technologies has revolutionized oncological studies to elucidate translational regulatory mechanisms. Ribosome profiling can provide adequate information on diverse aspects of translation by aiding in quantitatively analyzing the intensity of translating ribosome-protected fragments. Here, we review the primary currently used translatomics techniques and highlight their advantages and disadvantages as tools for translatomics studies. Subsequently, we clarified the areas in which ribosome profiling could be applied to better understand translational control. Finally, we summarized the latest advances in cancer studies using ribosome profiling to highlight the extensive application of this powerful and promising translatomic tool.

Abstract Osteoclasts (OCs), derived from monocyte/macrophage lineage, are key orchestrators in bone remodeling. Targeting osteoclast apoptosis is a promising approach to cut down excessive osteoclast numbers, and thus slow down the rate of bone mass loss that inevitably occurs during aging. However, the therapeutic target of apoptosis in osteoclasts has not been fully studied. Our previous work generated Mvp f/f Lyz2-Cre mice, conditionally depleting major vault protein (MVP) in monocyte lineage, and identified MVP as a bone protector for its negative role in osteoclastogenesis in vivo and in vitro. Here, we observed a notable decline of MVP in osteoclasts with aging in mice, encouraging us to further investigate the regulatory role of osteoclast MVP. Then, Mvp f/f Lyz2-Cre mice were exploited in two osteoporosis contexts, aging and abrupt loss of estrogen, and we revealed that conditional knockout of MVP inhibited osteoclast apoptosis in vivo and in vitro. Moreover, we reported the interaction between MVP and death receptor Fas, and MVP-Fas signaling cascade was identified to positively regulate the apoptosis of osteoclasts, thus preventing osteoporosis. Collectively, our comprehensive discovery of MVP’s regulatory role in osteoclasts provides new insight into osteoclast biology and therapeutic targets for osteoporosis.

Summary: Nuclear factor kappa B (NF-κB) plays a pivotal role in the development of pancreatic cancer, and its phosphorylation has previously been linked to the regulation of NUAK2. However, the regulatory connection between NF-κB and NUAK2, as well as NUAK2’s role in pancreatic cancer, remains unclear. In this study, we observed that inhibiting NUAK2 impeded the proliferation, migration, and invasion of pancreatic cancer cells while triggering apoptosis. NUAK2 overexpression partially resisted apoptosis and reversed the inhibitory effects of the NF-κB inhibitor. NF-κB transcriptionally regulated NUAK2 transcription by binding to the promoter region of NUAK2. Mechanistically, NUAK2 knockdown remarkably reduced the expression levels of p-SMAD2/3 and SMAD2/3, resulting in decreased nuclear translocation of SMAD4. In SMAD4-negative cells, NUAK2 knockdown impacted FAK signaling by downregulating SMAD2/3. Moreover, NUAK2 knockdown heightened the sensitivity of pancreatic cancer cells to gemcitabine, suggesting that NUAK2 inhibitors could be a promising strategy for pancreatic cancer treatment.

IntroductionDifenoconazole (DIFE) is a common pesticide used in citrus cultivation; excessive intake can cause neurological damage to the organism, and the existing colloidal gold immunochromatographic test strips cannot meet the requirements for the detection of citrus samples.MethodsDifenoconazole test strip was prepared based on the colloidal gold immunochromatographic technique (GICT), and its application in citrus samples was investigated; with colloidal gold (CG) as the probe, the optimization of GICT parameters, and the determination of reaction method, the immunochromatographic test strips for the detection of DIFE in citrus was developed, and the limit of detection (LOD), specificity, accuracy, and stability of the test strips were verified.ResultsThe results showed that the visual detection limit of the prepared colloidal gold immunochromatographic test strips was 0.2 mg/kg and the quantitative range was 0.06–0.6 mg/kg, and the test strips could specifically identify DIFE and have no cross-reaction with other common triazole pesticides. The detection method established in this study was verified by the GC–MS method, and the detection results achieved good consistency (R2 > 0.98).ConclusionThe test strips developed in this study have good performance and can be used for highly sensitive detection of citrus samples.

Living grass mulching (LMG) is a modern, environmentally friendly, practical, and efficient production management technology that improves the ecological environment, quality, and efficiency of the orchard. However, in arid and semi-arid areas, the effects of different grass species mulching on soil carbon composition, carbon pool stability, and nitrogen content are still unclear. Therefore, in order to explore the impact of different LMG on soil carbon, nitrogen, and its component content, as well as the related soil carbon pool management index in an apple orchard located in the semi-arid region of the Loess Plateau, a control experiment was conducted. The experiment involved different grass species cover treatments on an 11-year-old semi-dwarf Qinguan apple orchard from 2019 to 2022. Soil carbon and nitrogen content were measured under each treatment. The results indicated that the application of LMG treatment and depth of the soil had a significant impact on the soil organic carbon (SOC), particulate organic carbon (POC), inactive organic carbon (NAOC), total nitrogen (TN), and carbon-to-nitrogen ratio (C/N). Planting Vulpia myuros mulches significantly enhanced 39.6% surface soil organic carbon, 61.7% surface particulate organic carbon, 20.3% surface dissolved organic carbon (DOC), 75.8% surface inactive organic carbon, and 20.6% surface soil total nitrogen compared to clean tillage. Mulching treatment with the planting of Vulpia myuros boosted surface soil organic carbon and decreased soil carbon pool activity (CPA) and carbon pool activity index (CPAI), ultimately improving the stability of the soil carbon pool. The findings will have a beneficial impact on improving soil quality, carbon sequestration, and emission reduction in arid and semi-arid regions.

Environmental perception is a key indicator for evaluating green space preference and satisfaction. The shift in urban green space research towards emphasizing human benefits over functionality has caused a gradual increase in the number of articles on green space perception in recent years. Studying the relationship between the vegetation landscape (a fundamental aspect of urban green spaces) and subjective perceptual dimensions have become a pivotal research topic. This paper delves into the correlations and influencing relationships between these two dimensions with the main focus being on vegetation landscape indicators and subjective perception. The perceived green benefits were categorized into four main groups in the 80 articles reviewed: perceived safety, visual aesthetics, restoration benefits, and preferences. In this study, green indicators were also defined and categorized. The results indicated that the guiding indicators had a higher frequency and that species diversity improved the perceived benefits in all aspects. Among the control indicators, plant color, plant height, and the vegetation composition structure received the most attention. Plant color accounted for the largest proportion of studies on perceived visual aesthetics. Vegetation characteristics related to perceived safety had a shared trait, and there was a lack of correlation studies between safety and aesthetics in perceptual dimensions. In the common methods for assessing human perception, a shift from subjective measurements to mixed measurements using emerging technologies is underway. As a pioneering scope review of how vegetation landscapes influence subjective human perceptions, this study has not only provided evidence of the perceived benefits of urban green spaces but also offers valuable insights and tools for future research and policymaking.

BackgroundDiabetic kidney disease (DKD) has become the leading cause of kidney failure, causing a significant socioeconomic burden worldwide. The usual care for DKD fails to achieve satisfactory effects in delaying the persistent loss of renal function. A Chinese herbal medicine, Tangshen Qushi Formula (TQF), showed preliminary clinical benefits with a sound safety profile for people with stage 2-4 DKD. We present the protocol of an ongoing clinical trial investigating the feasibility, efficacy, and safety of TQF compared to placebo in delaying the progressive decline of renal function for people with stage 2-4 DKD.MethodsA mixed methods research design will be used in this study. A randomized, double-blind, placebo-controlled pilot trial will evaluate the feasibility, efficacy, and safety of TQF compared to placebo on kidney function for people with stage 2-4 DKD. An embedded semi-structured interview will explore the acceptability of TQF granules and trial procedures from the participant’s perspective. Sixty eligible participants with stage 2-4 DKD will be randomly allocated to the treatment group (TQF plus usual care) or the control group (TQF placebo plus usual care) at a 1:1 ratio for 48-week treatment and 12-week follow-up. Participants will be assessed every 12 weeks. The feasibility will be assessed as the primary outcome. The changes in the estimated glomerular filtration rate, urinary protein/albumin, renal function, glycemic and lipid markers, renal composite endpoint events, and dampness syndrome of Chinese medicine will be assessed as the efficacy outcomes. Safety outcomes such as liver function, serum potassium, and adverse events will also be evaluated. The data and safety monitoring board will be responsible for the participants’ benefits, the data’s credibility, and the results’ validity. The intent-to-treat and per-protocol analysis will be performed as the primary statistical strategy.DiscussionConducting a rigorously designed pilot trial will be a significant step toward establishing the feasibility and acceptability of TQF and trial design. The study will also provide critical information for future full-scale trial design to further generate new evidence supporting clinical practice for people with stage 2-4 DKD.Trial registration numberhttps://www.chictr.org.cn/, identifier ChiCTR2200062786.

Drought and nitrogen addition are important factors influencing soil microorganisms and changes in the soil environment. In the future, droughts will become more frequent, shorter, and more severe. However, little is known about the soil organic carbon components, enzyme activity, and composition, structure, and functional genes of soil microbial communities under short-term drought and nitrogen application conditions. In this study, we used metagenomics sequencing technology to explore the changes in the composition and functional genes of soil microbial communities under short-term drought, nitrogen application, and their interactions in the artificial grasslands of the Loess Plateau. The results indicated that (I) short-term drought, nitrogen application, and their interactions all increased the particulate organic carbon (POC) content. (II) Short-term drought increased the contents of soil microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN), as well as soil enzyme activity. (III) Short-term drought significantly increased the bacterial alpha diversity, whereas the interactions of short-term drought and nitrogen application enhanced the fungal alpha diversity. (IV) The interactions of short-term drought and nitrogen application inhibited Fusarium to prevent plant diseases. (V) Short-term drought enriched the relative abundance of genes related to carbon cycling and amino acid metabolism, while nitrogen application reduced genes related to carbon cycling but enriched genes related to glycan biosynthesis and metabolism. These results clearly showed that, short-term drought altered the composition and functional genes of soil microbial communities. Our research suggests that in the event of frequent short-term droughts in the future, nitrogen addition can be considered to maintain the diversity of soil microbial communities and sustain soil carbon and nitrogen cycling.

Purpose: To develop and validate a deep learning model that can transform color fundus (CF) photography into corresponding venous and late-phase fundus fluorescein angiography (FFA) images. Design: Cross-sectional study. Participants: We included 51 370 CF-venous FFA pairs and 14 644 CF-late FFA pairs from 4438 patients for model development. External testing involved 50 eyes with CF-FFA pairs and 2 public datasets for diabetic retinopathy (DR) classification, with 86 952 CF from EyePACs, and 1744 CF from MESSIDOR2. Methods: We trained a deep-learning model to transform CF into corresponding venous and late-phase FFA images. The translated FFA images’ quality was evaluated quantitatively on the internal test set and subjectively on 100 eyes with CF-FFA paired images (50 from external), based on the realisticity of the global image, anatomical landmarks (macula, optic disc, and vessels), and lesions. Moreover, we validated the clinical utility of the translated FFA for classifying 5-class DR and diabetic macular edema (DME) in the EyePACs and MESSIDOR2 datasets. Main Outcome Measures: Image generation was quantitatively assessed by structural similarity measures (SSIM), and subjectively by 2 clinical experts on a 5-point scale (1 refers real FFA); intragrader agreement was assessed by kappa. The DR classification accuracy was assessed by area under the receiver operating characteristic curve. Results: The SSIM of the translated FFA images were > 0.6, and the subjective quality scores ranged from 1.37 to 2.60. Both experts reported similar quality scores with substantial agreement (all kappas > 0.8). Adding the generated FFA on top of CF improved DR classification in the EyePACs and MESSIDOR2 datasets, with the area under the receiver operating characteristic curve increased from 0.912 to 0.939 on the EyePACs dataset and from 0.952 to 0.972 on the MESSIDOR2 dataset. The DME area under the receiver operating characteristic curve also increased from 0.927 to 0.974 in the MESSIDOR2 dataset. Conclusions: Our CF-to-FFA framework produced realistic FFA images. Moreover, adding the translated FFA images on top of CF improved the accuracy of DR screening. These results suggest that CF-to-FFA translation could be used as a surrogate method when FFA examination is not feasible and as a simple add-on to improve DR screening. Financial Disclosure(s): Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Background: Pancreatic fistula (PF) and biliary fistula (BF) are two major leakage complications after pancreatoduodenectomy (PD). The aim of this study is to investigate the risk factors of PF and BF after laparoscopic PD (LPD). Materials and Methods: We conducted a retrospective analysis of 500 patients who underwent LPD from 1 April 2015 to 31 March 2020. Clinical data from patients were analysed using multivariate logistic regression analysis. Results: PF occurred in 86 (17.2%) patients. Univariate and multivariate analysis indicated that the soft texture of the pancreas (P = 0.001) was the independent risk factor for PF. BF occurred in 32 (6.4%) patients. Univariate and multivariate analysis indicated that history of cardiovascular disease (P < 0.001), surgical time (P = 0.005), pre-operative CA125 (P = 0.036) and pre-operative total bilirubin (P = 0.044) were independent risk factors for BF. Conclusion: The texture of the pancreas was an independent risk factor for PF after LPD, which was consistent with the literatures. In addition, history of cardiovascular disease, surgical time, pre-operative CA125 and pre-operative total bilirubin were new independent risk factors for BF after LPD. Therefore, patients with high-risk factors of BF should be informed that they are at a high risk for this complication.

Abstract Metastasis constitutes one of the major causes of tumor-related death in gastric cancer (GC), and understanding key events in the initiation of this phenotypic switch may provide therapeutic opportunities. Long noncoding RNAs (lncRNAs) are emerging as molecules that play vital roles in tumorigenesis and metastasis. In this study, we aimed to identify metastasis-related lncRNAs in the context of GC. The lncRNAs overexpressed in tumor tissues and positively associated with overall survival were screened out using the TCGA database. qPCR assays in clinical samples showed that LINC00858 was significantly upregulated in GC tissues compared with normal counterparts. Functional analysis suggested that LINC00858 depletion attenuated the migration, and invasion of cancer cells in vitro and suppressed the metastasis of xenografted tumors in vivo. Mechanistically, LINC00858 could interact with the metastasis-associated RAN and stabilize its protein expression by decreasing posttranslational ubiquitination. The transcription factor YY1 could bind to the promoter of LINC00858 to upregulate its expression in GC cells. Moreover, overexpression of YY1 and RAN was positively associated with upregulation of LINC00858 in GC tissues. Our results suggest that LINC00858 might play a role in GC metastasis, and be a diagnostic biomarker and potential therapeutic target.

BackgroundThe health of office workers has become a major concern under the pressure of increasingly fierce job competition. As countries have gradually promoted healthy buildings, there is an urgent need to create and construct healthy office environments. Although the WELL Building Standard proposed management and design strategies based on the principles of health and medicine, it does not consider group characteristics or gender differences.PurposeThis study aims to apply the theory of planned behavior to healthy building design and supplement the important role of gender and group characteristics in behavioral guidance based on architectural strategies and user behaviors to improve the relevant building evaluation system.MethodsThis study adopted a questionnaire survey and structural equation model. Four WELL-certified healthy office buildings in Nanshan District, Shenzhen, were selected for the survey. Based on the theory of planned behavior, structural equation models for men and women were established, compared, and analyzed. The factors affecting the health behaviors of the two groups and the actual effectiveness of various building optimization strategies were discussed, and an optimization direction for gender differences was proposed.ResultsThe findings indicated differences between male and female staff in their individual characteristics and implementation of health behaviors. Management strategies, subjective design strategies in assistance and guidance, and objective design strategies in spatial planning can promote the health behaviors of the two groups. However, the design strategies of result feedback and detail optimization only appeared to have a significant positive effect on female staff, whereas the intelligent automation design strategies only had an obvious intervention effect on men’s health behaviors.SignificanceThis study found that the theory of planned behavior in the field of social psychology could be applied to relevant research on architectural design and emphasized the influence of gender. It can not only provide the optimization direction for the evaluation standards of relevant healthy buildings but also promote the implementation of health behaviors in office groups and provide new ideas for promoting the development of healthy buildings.

Background: Right ventricular involvement in hypertrophic cardiomyopathy is uncommon. This study aimed to evaluate clinical outcomes of the modified septal myectomy in patients diagnosed with biventricular hypertrophic cardiomyopathy (BHCM), a subject seldom explored in the literature. Methods: We conducted a retrospective cohort study from January 2019 to January 2023, enrolling 12 patients with BHCM. Each patient underwent a modified septal myectomy and was followed postoperatively. Clinical data and echocardiographic parameters, including the ventricular outflow tract peak pressure gradient and maximum interventricular septum thickness, were collected and analyzed. Results: The study cohort had a median age of 43.0 (interquartile range 14.5–63.0) years at surgery, with four patients (33.3%) being children. Two patients (16.7%) previously underwent percutaneous transluminal septal myocardial ablation. Surgical relief of biventricular outflow tract obstruction (BVOTO) was achieved in five patients (41.7%), aside from those managed solely for left ventricular outflow tract obstruction. In five instances, three-dimensional (3D) printing technology assisted in surgical planning. The postoperative interventricular septum thickness was significantly reduced (21.0 mm preoperative vs. 14.5 mm postoperative, p < 0.001), effectively eliminating residual ventricular outflow tract obstruction. There were no severe complications, such as septal perforation or third-degree atrioventricular block. During a mean follow up of 21.2 ± 15.3 months, no sudden deaths, residual outflow tract obstruction, permanent pacemaker implantation, recurrent systolic anterior motion, or reoperations were reported. Conclusions: Our findings affirm that the modified septal myectomy remains the gold standard treatment for BHCM, improving patient symptoms and quality of life. BVOTO relief can be safely and effectively achieved through septal myectomy via transaortic and pulmonary valve approaches in selected patients. For intricate cases, the application of 3D printing technology as a preoperative planning tool is advised to optimize surgical precision and safety.

Elevated temperature and frequent drought events under global climate change may seriously affect soil respiration. However, the underlying mechanism of the effects of warming and drought on soil respiration is not fully understood in the context of the Loess Plateau. This study examined the response of soil respiration (Rs) to multiple factors, including warming (W), drought (P), and their interaction (WP), in the semi-arid grassland of the Loess Plateau in Northwest China. The research period was from May to November 2022, with an open-top heating box used for warming and a rain shelter used for drought. The results showed the following: (1) Rs ranged from 1.67 μmol m−2 s−1 to 4.77 μmol m−2 s−1, with an average of 3.36 ± 0.07 μmol m−2 s−1. The cumulative soil carbon flux ranged from 500.97 g C·m−2 to 566.97 g C·m−2, and the average cumulative soil respiration was 535.28 ± 35.44 g C·m−2. (2) Warming increased Rs by 5.04 ± 3.11%, but drought inhibited Rs by 3.40 ± 3.14%, and the interaction between warming and drought significantly reduced soil respiration by 11.27 ± 3.89%. (3) The content of particulate organic carbon (POC), dissolved organic carbon (DOC), soil organic carbon (SOC), and readily oxidized carbon (ROC) decreased with the increased soil depth. ROC after W and WP treatments was significantly higher than that of the control, and POC after P treatment was significantly higher than CK (p < 0.05). (4) The seasonal variation of soil respiration was positively correlated with soil temperature, soil water content, plant height, and leaf area index (p < 0.05), but the response rules differed during different regeneration periods. Soil water content; soil water content and leaf area index; and soil water content, soil temperature, and leaf area index were the factors that regulated the variation in soil respiration in the first, second, and third regeneration periods, respectively. These results clearly showed the limiting effect of drought stress on the coupling between temperature and soil respiration, especially in semi-arid regions. Collectively, the variations in soil respiration under warming, drought, and their interactions were further regulated by different biotic and abiotic factors. Considering future warming, when coupled with increased drought, our findings indicate the importance of considering the interactive effects of climate change on soil respiration and its components in arid and semi-arid regions over the next decade.

Gene bla NDM, conferring resistance to carbapenem antibiotics, is globally distributed across Gram-negative bacteria on multiple plasmids. Here, Acman et al. study the dynamics underlying bla NDM dissemination across over 6000 bacterial genomes, and identify mobile genetic elements and specific mobilisation events likely involved in the gene’s global spread.

Hardware electronic synapse and neuro-inspired computing system based on phase change random access memory (PCRAM) have attracted an extensive investigation. However, due to the intrinsic asymmetric reversible phase transition, the defective weight update of PCRAM synapses in aspects of tuning range, linearity and continuity has long required a system-level complexity of circuits and algorithms. The cell-level improvements to a great extent may slim the system thus achieving efficient computing. We report in this work the great enhancement of Ge2Sb2Te5 (GST) based PCRAM synapses by combining materials engineering and pulse programming. It is found that carbon doping in GST retards the rate of phase changing thus increasing the controllability of the conductance, while non-linear programmable pulse excitations can eventually lead to a reliable synaptic potentiation and depression. A set of improved programmable pulse schemes for spike-timing dependent plasticity was then demonstrated, suggesting its potential superiority in flexible programming and reliable data collection. Our methods and results are of great significance for implementing PCRAM electronic synapses and high-performance neuro-inspired computing.

Random numbers are widely used in numerical computing, statistical simulation, random sampling, etc. At present, the mechanism for generating random numbers by computers is at risk of being attacked. The generated random numbers may be predicted in some cases,. However, current RNGs used in blockchain are not sufficient enough to handle attacks. Therefore, it is necessary to improve the security of random numbers. This paper improves the random number generator and designs a secure random number generator based on blockchain (BCsRNG). In practice, an encapsulated function algorithm of the secure random number is developed with the smart contract. Moreover, an API is provided to facilitate input and output. The experimental results show that the random numbers generated by BCsRNG are difficult to decipher and have higher randomness and security.

BackgroundIncreasing evidence shows that systemic inflammation is an embedded mechanism of proliferative diabetic retinopathy (PDR). However, the specific systemic inflammatory factors involved in this process remained obscure. The study aimed to identify the upstream and downstream systemic regulators of PDR by using Mendelian randomization (MR) analyses.MethodsWe performed a bidirectional two-sample MR analysis implementing the results from genome-wide association studies for 41 serum cytokines from 8,293 Finnish individuals, and PDR from FinnGen consortium (2,025 cases vs. 284,826 controls) and eight cohorts of European ancestry (398 cases vs. 2,848 controls), respectively. The inverse-variance-weighted method was adopted as the main MR method, and four additional MR methods (MR-Egger, weighted-median, MR-pleiotropy residual sum and outlier (MR-PRESSO), and MR-Steiger filtering methods) were used for the sensitivity analyses. Results from FinnGen and eight cohorts were pooled into a meta-analysis.ResultsOur results showed that genetically predicted higher stem cell growth factor-β (SCGFb) and interleukin-8 were positively associated with an elevated risk of PDR, with a combined effect of one standard deviation (SD) increase in SCGFb and interleukin-8 causing 11.8% [95% confidence interval (CI): 0.6%, 24.2%]) and 21.4% [95% CI: 3.8%, 41.9%]) higher risk of PDR, respectively. In contrast, genetically predisposition to PDR showed a positive association with the increased levels of growth-regulated oncogene-α (GROa), stromal cell-derived factor-1 alpha (SDF1a), monocyte chemotactic protein-3 (MCP3), granulocyte colony-stimulating factor (GCSF), interleukin-12p70, and interleukin-2 receptor subunit alpha (IL-2ra).ConclusionsOur MR study identified two upstream regulators and six downstream effectors of PDR, providing opportunities for new therapeutic exploitation of PDR onset. Nonetheless, these nominal associations of systemic inflammatory regulators and PDR require validation in larger cohorts.

ABSTRACT Carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) is recognized as a threat worldwide, but the mechanisms underlying its emergence remain unclear. As most CR-hvKP isolates are not hypermucoviscous, we speculated that the evolution of the capsule might result in the convergence of carbapenem resistance and hypervirulence. Here, 2,096 K. pneumoniae isolates were retrospectively collected to screen the ST23-K1 clone, and hypervirulence was roughly defined as being highly resistant to serum killing. The effect of wcaJ on the capsule, virulence, fitness, and resistance acquisition was further analyzed. The capsule gene wcaJ, inserted by ISKpn26/ISKpn74, was identified via whole-genome sequencing in four hvKP, but not hypermucoviscous, isolates. Uronic acid quantitation results revealed that these isolates produced significantly less capsular polysaccharides than NTUH-K2044. A significant increase in capsular production was observed in wcaJ-complemented isolates and confirmed by transmission electron microscopy. Further, all wcaJ-complemented isolates acquired greater resistance to macrophage phagocytosis, and one representative isolate resulted in a significantly higher mortality rate than the parental isolate in mice, indicating that wcaJ inactivation might compromise virulence. However, isolates with wcaJ interruption demonstrated a lower fitness cost and a high conjugation frequency of the blaKPC-2 plasmid, raising concerns about the emergence of carbapenem resistance in hvKP. IMPORTANCE Klebsiella pneumoniae is one of the most common nosocomial pathogens worldwide, and we speculated that the evolution of the capsule might result in the convergence of carbapenem resistance and hypervirulence of K. pneumoniae. The wcaJ gene was first reported to be interrupted by insertion sequence elements in ST23-K1 hypervirulent Klebsiella pneumoniae, resulting in little capsule synthesis, which plays an important role in virulence. We examined the effect of wcaJ on the capsule, virulence, and fitness. Isolates with wcaJ interruption might compromise virulence and demonstrated a lower fitness cost and a high conjugation frequency of the blaKPC-2 plasmid, highlighting its role as a potential factor facilitating hypervirulence and carbapenem resistance.

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) of the sequence type 59 (ST59) and ST398 lineages has emerged in hospitals and displayed a higher virulent potential than its counterparts ST5 and ST239. However, the mechanism of the host cell-pathogen interaction and specific determinates that contribute to the success of epidemic clones remain incompletely understood. In the present study, 142 S. aureus strains (ST59, ST398, ST239, and ST5) were selected from our 7-year national surveillance of S. aureus bloodstream infections (n = 983). We revealed that ST59 and ST398 had a higher prevalence of the protease-associated genes hysAVSaβ, paiB, and cfim and enhanced proteolytic activity than the other lineages. ST59 and ST398 showed a higher expression of RNAIII and psmα and greater proficiency at causing cell lysis than other lineages. Furthermore, ST59 and ST398 were strongly recognized by human neutrophils and caused more cell apoptosis and neutrophil extracellular trap degradation than the other lineages. In addition, these strains differed substantially in their repertoire and composition of intact adhesion genes. Moreover, ST398 displayed higher adaptability to human epidermal keratinocytes and a unique genetic arrangement inside the oligopeptide ABC transport system, indicating functional variations. Overall, our study revealed some potential genomic traits associated with virulence and fitness that might account for the success of epidemic clones. IMPORTANCE Considerable efforts have been exerted to identify factors contributing to the success of epidemic Staphylococcus aureus clones, however, comparative phenotypic studies lack representation owing to the small number of strains. Large-scale strain collections focused on the description of genomic characteristics. Moreover, methicillin-resistant S. aureus infections constitute 30% to 40% of S. aureus bloodstream infections, and recent research has elucidated highly virulent methicillin-susceptible S. aureus strains. However, comprehensive research on the factors contributing to the success of epidemic S. aureus clones is lacking. In this study, 142 S. aureus strains were selected from our 7-year national surveillance of S. aureus bloodstream infections (n = 983) accompanied by a rigorous strain selection process. A combination of host cell-pathogen interactions and genomic analyses was applied to the represented strains. We revealed some potential genomic traits associated with virulence and fitness that might account for the success of epidemic clones.

Abstract Phase‐change memory (PCM) has been developed for three‐dimensional (3D) data storage devices, posing huge challenges to the thermal stability and reliability of PCM. However, the low thermal stability of Ge2Sb2Te5 (GST) limits further application. Here, we demonstrate PCM based on In0.9Ge2Sb2Te5 (IGST) alloy, showing 180°C 10‐years data retention, 6 ns set speed, one order of magnitude longer life time, and 75% reduced power consumption compared to GST‐based device. The In can occupy the cationic positions and the In‐Te octahedrons with good phase‐change properties can geometrically match well with the host Ge‐Te and Sb‐Te octahedrons, acting as nucleation centers to boost the set speed and enhance the endurance of IGST device. Introducing stable matched phase‐change octahedrons can be a feasible way to achieve practical PCMs.

AimsThis study aimed to investigate the prognostic value of clinical features for cancer-specific survival (CSS) and metastasis in patients with pancreatic mucinous cystadenocarcinoma (MCAC). We further constructed and validated an effective nomogram to predict CSS.MethodsWe screened patients diagnosed with pancreatic MCAC from Surveillance Epidemiology and End Results (SEER) database. Kaplan-Meier curves were used to determine the CSS time. Univariate and multivariate Cox and logistic regression analyses were conducted to identify the prognostic factors for CSS and metastasis. The nomogram was constructed to predict the prognosis of pancreatic MCAC based on the results from the multivariate analysis. We used the concordance index (C-index), the area under the curve (AUC), and the calibration plots to determine the predictive accuracy and discriminability of the nomogram.ResultsMultivariate Cox analysis revealed that age, primary site, grade, and radiotherapy were independent prognostic factors associated with CSS. Multivariate logistic regression analysis revealed that surgery and grade were independent risk factors associated with metastasis. The independent risk factors were included to construct a prognosis prediction model for predicting CSS in patients with pancreatic MCAC. The concordance index (C-index), receiver operating characteristic (ROC) curves, and calibration plots of the training cohort and the validation cohort showed that the nomogram had an acceptable predictive performance.ConclusionWe established a nomogram that could determine the 3- and 5-year CSS, which could evaluate individual clinical outcomes and provide individualized clinical decisions.

Abstract Nonvolatile phase‐change random access memory (PCRAM) is regarded as one of the promising candidates for emerging mass storage in the era of Big Data. However, relatively high programming energy hurdles the further reduction of power consumption in PCRAM. Utilizing narrow edge‐contact of graphene can effectively reduce the active volume of phase change material in each cell, and therefore realize low‐power operation. Here, it demonstrates that the power consumption can be reduced to ≈53.7 fJ in a cell with ≈3 nm‐wide graphene nanoribbon (GNR) as edge‐contact, whose cross‐sectional area is only ≈1 nm2. It is found that the polarity of the bias pulse determines its cycle endurance in the asymmetric structure. If a positive bias is applied to the graphene electrode, the endurance can be extended at least one order longer than the case with a reversal of polarity. In addition, the introduction of the hexagonal boron nitride (h‐BN) multilayer leads to a low resistance drift and a high programming speed in a memory cell. The work represents a great technological advance for the low‐power PCRAM and can benefit in‐memory computing in the future.

ABSTRACT Cefiderocol has been approved in the United States and Europe but not in China. We aim to evaluate carbapenem-resistant Enterobacterales (CRE) susceptibility to cefiderocol to provide baseline data and investigate the resistance mechanism. From 2018 to 2019, 1,158 CRE isolates were collected from 23 provinces and municipalities across China. The MICs of antimicrobials were determined via the agar dilution and broth microdilution methods. Whole-genome sequencing was performed for 26 cefiderocol-resistant Escherichia coli isolates to investigate the resistance mechanism. Clone transformations were used to explore the function of cirA, pbp3, and blaNDM-5 in resistance. Among the 21 antimicrobials tested, aztreonam-avibactam had the highest antibacterial activity (98.3%), followed by cefiderocol (97.3%) and colistin (95.3%). A total of 26 E. coli isolates harboring New Delhi metallo-beta-lactamase 5 (NDM-5) showed high levels of cefiderocol resistance, of which sequence type 167 (ST167) accounted for 76.9% (20/26). We found 4 amino-acid insertions (YRIN/YRIK) at position 333 of penicillin-binding protein 3 (PBP3) in the 26 E. coli isolates, and 22 isolates had a siderophore receptor cirA premature stop codon. After obtaining the wild-type cirA supplementation, the MIC of the transformants decreased by 8 to 16 times in two cefiderocol-resistant isolates. A cefiderocol-susceptible isolate harboring NDM-5 has an MIC increased from 1 μg/mL to 64 μg/mL after cirA deletion, and the MIC decreased from 64 μg/mL to 0.5 μg/mL after blaNDM-5 deletion. The MIC of the E. coli DH5α, from which the pbp3 mutant was obtained, increased from 0.064 μg/mL to 0.25 μg/mL. Cefiderocol showed activity against most CRE in China. The resistance of ST167 E. coli to cefiderocol is a combination of the premature stop codon of cirA, pbp3 mutation, and blaNDM-5 existence. IMPORTANCE Cefiderocol, a new siderophore cephalosporin, has been approved in the United States and Europe but not in China. At present, there are almost no antimicrobial susceptibility evaluation data on cefiderocol in China. We evaluated the in vitro susceptibility of 1,158 strains of carbapenem-resistant Enterobacterales to cefiderocol and other antibiotics. We found that a high proportion of Escherichia coli showed high-level resistance to cefiderocol. Whole-genome sequencing (WGS) and molecular cloning experiments confirmed that the synergistic effect of the cirA gene premature stop codon, blaNDM-5 existence, and the pbp3 mutation is associated with high levels of cefiderocol resistance.

ABSTRACT Carbapenem-resistant Acinetobacter baumannii (CRAB) is an opportunistic pathogen that has become a global threat. The dissemination of global clone 2 (GC2) CRAB has been well documented. Oxford sequence type (ST) 208 is one of the most prevalent lineages of A. baumannii GC2; however, its evolution and phylogeny are unclear. We collected 45 representative ST208 isolates from 14 cities in China between 1999 and 2018. Moreover, 411 ST208 genome sequences were downloaded from the GenBank database for comparison. The global ST208 phylogeny showed that ST208 might have originated from North America and subsequently evolved into two clades. Notably, the widespread OXA-23-producing ST208 A. baumannii was correlated with the transposon structure and dynamics of replicative transposition, and the Tn2009 tandem structure of five copies of blaOXA-23 and potential circular intermediate of Tn2009 were first detected. Furthermore, 15 Chinese ST208 isolates carried GR25 pABTJ1-like plasmids, which contained blaOXA-23 and have only been found in China in the last decade. In conclusion, our work suggests that replicative transposition contributes to the evolution and transmission of OXA-23-producing ST208 A. baumannii and highlights the new challenges posed by the epidemiological surveillance of globally distributed clonal groups via whole genome sequencing. IMPORTANCE ST208 as one of the most prevalent lineages of CRAB has caused several difficult-to-treat infections and outbreaks around the world. However, few studies have focused on evaluating the genetic background differences of ST208 A. baumannii isolated from very distant geographic regions. A comprehensive genomic analysis of 456 clinical strains of ST208 A. baumannii from a wide temporal and geographic range was performed in this study. Moreover, the mechanisms leading to the horizontal transfer of blaOXA-23 in ST208 A. baumannii are poorly understood. We first describe experimental evidence of the potential circular intermediate of Tn2009, and the Tn2009 tandem structure of five copies of blaOXA-23 was first detected. The interbacterial transfer of genetic elements carrying resistance to last-line antibiotic carbapenems highlights the essential need to enhance epidemiologic surveillance.

Carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) have been reported in recent years across Asian countries and pose a serious threat to public health. Neutrophils represent the first line of defense against numerous infectious pathogens, such as CR-hvKP. Neutrophil extracellular traps (NETs) constitute one of the major antimicrobial defense mechanisms in neutrophils against invading pathogens, especially against hvKP. Interestingly, previous studies have demonstrated that patients with type 2 diabetes mellitus (T2D) display elevated levels of NETosis but are vulnerable to infections caused by hvKP. The discrepancy propels us to investigate the role of NETs in hvKP infections in the context of T2D. By utilizing a clinical-derived CR-hvKP strain and a combination of NETs complex detection, phagocytosis testing, NETs killing assay and immunofluorescence, and scanning electron microscope assays, we identified defective NETs-mediated killing of CR-hvKP strain in patients with T2D. Specifically, we show that the impaired NETs-mediated killing in T2D is not due to the decreased NETs formation, as the neutrophils isolated from T2D patients exhibited enhanced NETs formation compared to healthy controls. Further, we demonstrate that the reduced NETs activity does not result from the trapping failure of CR-hvKP, but likely associated with the deficient surface damage conferred by the NETs of T2D patients. Our data provide a novel insight into the defective innate immune response against CR-hvKP in T2D.

High density phase change memory array requires both minimized critical dimension (CD) and maximized process window for the phase change material layer. High in-wafer uniformity of the nanoscale patterning of chalcogenides material is challenging given the optical proximity effect (OPE) in the lithography process and the micro-loading effect in the etching process. In this study, we demonstrate an approach to fabricate high density phase change material arrays with half-pitch down to around 70 nm by the co-optimization of lithography and plasma etching process. The focused-energy matrix was performed to improve the pattern process window of phase change material on a 12-inch wafer. A variety of patternings from an isolated line to a dense pitch line were investigated using immersion lithography system. The collapse of the edge line is observed due to the OPE induced shrinkage in linewidth, which is deteriorative as the patterning density increases. The sub-resolution assist feature (SRAF) was placed to increase the width of the lines at both edges of each patterning by taking advantage of the optical interference between the main features and the assistant features. The survival of the line at the edges is confirmed with around a 70 nm half-pitch feature in various arrays. A uniform etching profile across the pitch line pattern of phase change material was demonstrated in which the micro-loading effect and the plasma etching damage were significantly suppressed by co-optimizing the etching parameters. The results pave the way to achieve high density device arrays with improved uniformity and reliability for mass storage applications.

The negative effects of long working hours and shift work on the physical and mental health of blue-collar groups should not be underestimated. Under intense stress, they have limited time to access recreational green space, and their own health is thus affected. In this study, a conceptual model of recreational satisfaction among blue-collar workers was established based on the American Customer Satisfaction Index (ACSI). The model explores the factors affecting their level of satisfaction when using recreational spaces. Anning Industrial Park in Yunnan Province was used as an example. User data were collected and analyzed using a questionnaire survey and structural equation methods. The results indicate that recreation intention, perceived park quality characteristics, and perceived value all had significant and positive effects on the park recreation satisfaction of blue-collar workers. This study applied social economics theory to the field of landscape planning, identified the typical characteristics of blue-collar workers and their green space use, and strived to optimize the UGS configuration and functional facilities through the evaluation of recreation satisfaction indexes, which provided guidance and reference for improving the service quality of green spaces.

ABSTRACT Carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) poses a significant public health challenge worldwide, but research on IMP-producing CR-hvKP and its tigecycline resistance is extremely scarce. We report herein the recovery of two IMP-4–producing, capsular serotype K2, sequence type 65 (K2-ST65), hypervirulent K. pneumoniae isolates (C1672 and C2051), which caused severe and fatal infections in ICU patients, after retrospectively screening 3,285 carbapenem-resistant K. pneumoniae isolates from 26 provinces in China. Notably, C2051 also demonstrated tigecycline nonsusceptibility, mediated by a frameshift mutation in the TetR/AcrR family transcriptional regulator. Both strains harbored blaIMP-4 and critical plasmid-borne virulence genes (rmpA/rmpA2, iucA, and iroN) and demonstrated high virulence in Galleria mellonella, indicating CR-hvKP. The blaIMP-4 gene was located on the IncU- and IncN-type plasmids, which showed high stability in C1672 and C2051 after serial passage for 5 days, with retention rates of 87% and 93.7%, respectively. No significant differences in growth rates were observed among the parental strains and the corresponding resistance plasmid-cured mutants (P = 0.5273), suggesting that strains carrying the blaIMP and virulence plasmids have the potential to exist for a long time without compromising fitness. The genetic environments of the blaIMP-4 gene in both strains were similar, and it has been inferred that the genetic regions of blaIMP-4 were inserted into different backbones. Several conjugal transfer genes, such as traO, traE, traN, and traBCD, were identified in the blaIMP-4-bearing plasmid of C2051, suggesting a higher ability for plasmid transmission. The convergence of IMP carbapenemase and tigecycline nonsusceptibility in a classic hypervirulent K. pneumoniae lineage highlights the need to enhance clinical awareness and epidemiologic surveillance. IMPORTANCE To date, research on IMP-producing CR-hvKP is extremely scarce. Only one case of urinary tract infection caused by an IMP-6–producing K1-ST23 hypervirulent K. pneumoniae isolate in Japan was recorded, with a limited description of clinical information and genomic features. None of the published studies examined the virulence of the reported strains or the stability and fitness of resistance plasmids or presented a phylogenetic analysis. This dearth of data is notable because CR-hvKP infections are increasingly identified, but critical characteristics of the emerging resistance mediated by IMP carbapenemases in CR-hvKP remain unknown. Here, we report the emergence of two IMP-4 carbapenemase-producing K2-ST65 hypervirulent K. pneumoniae isolates that caused severe and fatal infections in clinical settings in China. Notably, one of them also demonstrated tigecycline nonsusceptibility. These strains carrying blaIMP and virulence plasmids had the potential to exist for a long time without compromising their fitness, highlighting the urgent need to enhance clinical awareness and epidemiologic surveillance to prevent their dissemination.

ABSTRACTTigecycline is considered one of the last-resort antimicrobials for carbapenem-resistant K. pneumoniae. Plasmid-mediated tigecycline resistance remains largely unclear. Here, by utilizing whole genome sequencing, we report a plasmid-mediated tigecycline resistance mechanism, a 6,489 bp Resistance-nodulation-division family (RND) efflux pump (tmexCD1-toprJ1 pump), that confers transferable tigecycline resistance in K pneumoniae isolated from patients and chickens. In addition, we identified high prevalence of the plasmids co-harbouring both tmexCD1-toprJ1 pump and mcr (tmexCD1-mcr co-harbouring plasmid) from human in our nationwide collection. Even worse, the tmexCD1-toprJ1 and mcr co-harbouring plasmid was also co-existed with blaNDM-harbouring IncX3 plasmid in the same host, resulting in pandrug resistance. Phylogenetic analysis suggested that the plasmid-borne tmexCD1-toprJ1 originated from the chromosome of Aeromonas spp. through Tn5393-mediating translocation. Both plasmid-harbored tmexCD1-toprJ1 gene and mcr-8 likely originated from animal isolates and then spread to human. Our findings highlight a substantial threat of tmexCD1-toprJ1-mcr8 co-harbouring IncFIA/IncFII plasmid to public health due to their mobile resistance to both tigecycline and colistin, emphasizing an urgent need for further global surveillance on this plasmid.

Abstract Background In China multidrug-resistant bacteria pose a considerable threat to public health. Antimicrobial resistance has weakened the effectiveness of many medicines widely used today. Thus, discovering new antibacterial drugs is paramount in the effort to treat emerging drug-resistant bacteria. Methods Eravacycline, tigecycline and other clinical routine antibiotics were tested by reference broth micro-dilution method against 336 different strains collected from 11 teaching hospitals in China between 2012 and 2016. These isolates included Enterobacteriaceae, non-fermentative, Staphylococcus spp., Enterococcus, and a number of fastidious organisms. The strains involved in this study possess the most important drug resistance characteristics currently known in China. Drug resistant bacteria such as those producing extended spectrum β-lactamases (ESBL) and carbapenemases (KPC-2 and NDM-1), and those exhibiting colistin resistance (mcr-1) and tigecycline were included in this study. Additionally, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), β-lactamase positive Haemophilus influenzae, and penicillin resistant Streptococcus pneumoniae (PRSP) were also included. Results Eravacycline exhibited good efficacy against all the strains tested, especially for organisms with ESBLs, carbapenemases, and mcr-1 gene compared with tigecycline and other antibiotics tested. The MIC values of eravacycline against carbapenemase producing Enterobacteriaceae and OXA-23-producing A. baumannii were much lower than the MIC values of other antibiotics. MRSA, VRE, β-lactamase positive Haemophilus influenza, and PRSP were sensitive to eravacycline in every strain tested. Furthermore, in most strains tested, the MICs of eravacycline were two to four-fold lower than the MICs of tigecycline. Conclusions Eravacycline has shown potent antibacterial activity against common and clinically important antibiotic-resistant pathogens. The MIC distribution of eravacycline was generally lower than that of tigecycline which demonstrates that this new drug is potentially more effective than the existing medications.

The nosocomial spread of carbapenem-resistant Enterobacterales (CRE) is extremely common, resulting in severe burdens on healthcare systems. In particular, the high-risk Klebsiella pneumoniae ST11 strain has a wide endemic area in China. The current study describes the results of continuous monitoring of CRE genotypes and phenotypes in a tertiary hospital in North China from 2012 to 2020. A total of 160 isolates were collected, including 109 Klebsiella. pneumoniae (68.13%), 29 Escherichia coli (26.60%), 12 Enterobacter cloacae (7.50%), and 10 other strains (6.25%). A total of 149 carbapenemase genes were detected, of which blaKPC-2 (51.0%) was the most common, followed by blaNDM-1 (22.82%), and blaNDM-5 (23.49%). Based on multi-locus sequence typing, the ST11 strain (66.1%) dominates K. pneumoniae, followed by ST15 (13.8%). Interestingly, the proportion of blaNDM (22.2%, 16/72) in ST11 K. pneumoniae was significantly increased in 2018–2019. Hence, whole-genome sequencing was performed on ST11 K. pneumoniae. Growth curves and in vitro competition experiments showed that K. pneumoniae carrying blaNDM exhibited a stronger growth rate (p < 0.001) and competition index (p < 0.001) than K. pneumoniae carrying blaKPC. Moreover, K. pneumoniae carrying blaNDM had a stronger biofilm-forming ability than K. pneumoniae carrying blaKPC (t = 6.578; p < 0.001). K. pneumoniae carrying blaKPC exhibited increased defense against bactericidal activity than K. pneumoniae carrying blaNDM. Thus, ST11 K. pneumoniae carrying blaNDM has strong adaptability and can locally replace K. pneumoniae carrying blaKPC to become an epidemic strain. Based on these findings, infection control and preventive measures should focus on the high-risk ST11-K. pneumoniae strain.

In recent years, semi-outdoor space has become an important research subject in the field of thermal comfort. Overhead space located on the ground floor is a common type of semi-outdoor space in China’s Lingnan region with a hot and humid climate. Its thermal comfort has been scarcely studied. This study aims to reveal the importance and influencing factors of overhead public spaces in hot and humid areas, and to explore the corresponding adaptive behaviors of people. In this research, several overhead public spaces in Shenzhen University were selected to conduct field measurements and questionnaire surveys (n = 509) in hot and cold seasons. The results indicated that the acceptable physiologically equivalent temperature (PET) range for 90% of the population was 26.2–30.4 °C in hot season, 9.9–19.2 °C in cold season, and 17.6–25.3 °C for the whole year. The respondents preferred “neutral” in cold season and “slightly cool” in hot season. Respondents in hot season were more eager to adjust their thermal comfort, while those in cold season were more comfortable exposing themselves to the sun. Concurrently, the neutral temperature and neutral temperature range for different seasons was obtained and compared with the results of other studies. The results provide references for thermal comfort adjustment in hot and humid areas as well as optimization suggestions for the planning and design of overhead spaces.

Background: Bloodstream infection (BSI) is one of the most common serious bacterial infections worldwide and also a major contributor to in-hospital mortality. Determining the predictors of mortality is crucial for prevention and improving clinical prognosis in patients with nosocomial BSI.Methods: A nationwide prospective cohort study was conducted from 2007 until 2016 in 16 teaching hospitals across China. Microbiological results, clinical information, and patient outcomes were collected to investigate the pathogenic spectrum and mortality rate in patients with BSI and identify outcome predictors using multivariate regression, prediction model, and Kaplan–Meier analysis.Results: No significant change was observed in the causative pathogen distribution during the 10-year period and the overall in-hospital mortality was 12.83% (480/3,741). An increased trend was found in the mortality of patients infected with Pseudomonas aeruginosa or Acinetobacter baumannii, while a decreased mortality rate was noted in Staphylococcus aureus-related BSI. In multivariable-adjusted models, higher mortality rate was significantly associated with older age, cancer, sepsis diagnosis, ICU admission, and prolonged hospital stay prior to BSI onset, which were also determined using machine learning-based predictive model achieved by random forest algorithm with a satisfactory performance in outcome prediction.Conclusions: Our study described the clinical and microbiological characteristics and mortality predictive factors in patients with BSI. These informative predictors would inform clinical practice to adopt effective therapeutic strategies to improve patient outcomes.

In recent years, cable-driven parallel robots (CDPRs) have drawn more and more attention due to the properties of large workspace, large payload capacity, and ease of reconfiguration. In this paper, we present a kinematic and dynamic modeling and workspace analysis for a novel suspended CDPR which generates Schönflies motions. Firstly, the architecture of the robot is introduced, and the inverse and forward kinematic problems of the robot are solved through a geometrical approach. Then, the dynamic equation of the robot is derived by separately considering the moving platform and the drive trains. Based on the dynamic equation, the dynamic feasible workspace of the robot is determined under different values of accelerations. Finally, experiments are performed on a prototype of the robot to demonstrate the correctness of the derived models and workspace.

Carbapenemase-producing Enterobacterales have become a severe public health concern because of their rapidly transmissible resistance elements and limited treatment options. The most effective antimicrobial combinations against carbapenemase-producing Enterobacterales are currently unclear. Here, we aimed to assess the therapeutic effects of seven antimicrobial combinations (colistin-meropenem, colistin-tigecycline, colistin-rifampicin, colistin-erythromycin, meropenem-tigecycline, meropenem-rifampicin, and meropenem-tigecycline-colistin) against twenty-four carbapenem-producing Enterobacterales (producing blaKPC, blaNDM, coexisting blaNDM and blaIMP, and coexisting mcr-1/8/9 and blaNDM genes) and one carbapenem-susceptible Enterobacterales using the checkerboard assay, time-kill curves, and scanning electron microscopy. None of the combinations were antagonistic. The combination of colistin-rifampicin showed the highest synergistic effect of 76% (19/25), followed by colistin-erythromycin at 60% (15/25), meropenem-rifampicin at 24% (6/25), colistin-meropenem at 20% (5/25), colistin-tigecycline at 20% (5/25), and meropenem-tigecycline at 4% (1/25). The triple antimicrobial combinations of meropenem-tigecycline-colistin had a synergistic effect of 100%. Most double antimicrobial combinations were ineffective on isolates with coexisting blaNDM and blaIMP genes. Meropenem with tigecycline showed no synergistic effect on isolates that produced different carbapenemase genes and were highly resistant to meropenem (92% meropenem MIC ≥ 16 mg/mL). Colistin-tigecycline showed no synergistic effect on Escherichia coli producing blaNDM–1 and Serratia marcescens. Time-kill curves showed that antimicrobial combinations achieved an eradication effect (≥ 3 log10 decreases in colony counts) within 24 h without regrowth, based on 1 × MIC of each drug. The synergistic mechanism of colistin-rifampicin may involve the colistin-mediated disruption of bacterial membranes, leading to severe alterations in their permeability, then causes more rifampicin to enter the cell and induces cell death. In conclusion, the antimicrobial combinations evaluated in this study may facilitate the successful treatment of patients infected with carbapenemase-producing pathogens.

The recent plasmid-mediated spread of the mobilized colistin resistance gene mcr-1 poses a significant public health threat, requiring worldwide monitoring and surveillance. Here, Wang et al. compile and analyze a data set of 457 mcr-1-positive sequenced isolates to investigate the origin and global distribution of mcr-1.

Metalloproteins are a family of proteins characterized by metal ion binding, whereby the presence of these ions confers key catalytic and ligand-binding properties. Due to their ubiquity among biological systems, researchers have made immense efforts to predict the structural and functional roles of metalloproteins. Ultimately, having a comprehensive understanding of metalloproteins will lead to tangible applications, such as designing potent inhibitors in drug discovery. Recently, there has been an acceleration in the number of studies applying machine learning to predict metalloprotein properties, primarily driven by the advent of more sophisticated machine learning algorithms. This review covers how machine learning tools have consolidated and expanded our comprehension of various aspects of metalloproteins (structure, function, stability, ligand-binding interactions, and inhibitors). Future avenues of exploration are also discussed.