GNNGL-PPI: multi-category prediction of protein-protein interactions using graph neural networks based on global graphs and local subgraphs.

Most proteins exert their functions by interacting with other proteins, making the identification of protein-protein interactions (PPI) crucial for understanding biological activities, pathological mechanisms, and clinical therapies. Developing effective and reliable computational methods for predicting PPI can significantly reduce the time-consuming and labor-intensive associated traditional biological experiments. However, accurately identifying the specific categories of protein-protein interactions and improving the prediction accuracy of the computational methods remain dual challenges. To tackle these challenges, we proposed a novel graph neural network method called GNNGL-PPI for multi-category prediction of PPI based on global graphs and local subgraphs. GNNGL-PPI consisted of two main components: using Graph Isomorphism Network (GIN) to extract global graph features from PPI network graph, and employing GIN As Kernel (GIN-AK) to extract local subgraph features from the subgraphs of protein vertices. Additionally, considering the imbalanced distribution of samples in each category within the benchmark datasets, we introduced an Asymmetric Loss (ASL) function to further enhance the predictive performance of the method. Through evaluations on six benchmark test sets formed by three different dataset partitioning algorithms (Random, BFS, DFS), GNNGL-PPI outperformed the state-of-the-art multi-category prediction methods of PPI, as measured by the comprehensive performance evaluation metric F1-measure. Furthermore, interpretability analysis confirmed the effectiveness of GNNGL-PPI as a reliable multi-category prediction method for predicting protein-protein interactions.

Evolving ω-amine transaminase AtATA guided by substrate-enzyme binding free energy for enhancing activity and stability against non-natural substrates.

In the field of chiral amine synthesis, ω-amine transaminase (ω-ATA) is one of the most established enzymes capable of asymmetric amination under optimal conditions. However, the applicability of ω-ATA toward more non-natural complex molecules remains limited due to its low transamination activity, thermostability, and narrow substrate scope. Here, by employing a combined approach of computational virtual screening strategy and combinatorial active-site saturation test/iterative saturation mutagenesis strategy, we have constructed the best variant M14C3-V5 (M14C3-V62A-V116S-E117I-L118I-V147F) with improved ω-ATA from Aspergillus terreus (AtATA) activity and thermostability toward non-natural substrate 1-acetylnaphthalene, which is the ketone precursor for producing the intermediate (R)-(+)-1-(1-naphthyl)ethylamine [(R)-NEA] of cinacalcet hydrochloride, showing activity enhancement of up to 3.4-fold compared to parent enzyme M14C3 (AtATA-F115L-M150C-H210N-M280C-V149A-L182F-L187F). The computational tools YASARA, Discovery Studio, Amber, and FoldX were applied for predicting mutation hotspots based on substrate-enzyme binding free energies and to show the possible mechanism with features related to AtATA structure, catalytic activity, and stability in silico analyses. M14C3-V5 achieved 71.8% conversion toward 50 mM 1-acetylnaphthalene in a 50 mL preparative-scale reaction for preparing (R)-NEA. Moreover, M14C3-V5 expanded the substrate scope toward aromatic ketone compounds. The generated virtual screening strategy based on the changes in binding free energies has successfully predicted the AtATA activity toward 1-acetylnaphthalene and related substrates. Together with experimental data, these approaches can serve as a gateway to explore desirable performances, expand enzyme-substrate scope, and accelerate biocatalysis.IMPORTANCEChiral amine is a crucial compound with many valuable applications. Their asymmetric synthesis employing ω-amine transaminases (ω-ATAs) is considered an attractive method. However, most ω-ATAs exhibit low activity and stability toward various non-natural substrates, which limits their industrial application. In this work, protein engineering strategy and computer-aided design are performed to evolve the activity and stability of ω-ATA from Aspergillus terreus toward non-natural substrates. After five rounds of mutations, the best variant, M14C3-V5, is obtained, showing better catalytic efficiency toward 1-acetylnaphthalene and higher thermostability than the original enzyme, M14C3. The robust combinational variant acquired displayed significant application value for pushing the asymmetric synthesis of aromatic chiral amines to a higher level.

Efficacy of microwave ablation in the treatment of large benign thyroid nodules: a multi-center study.

OBJECTIVE: To evaluate the efficacy, safety, and improvement of symptoms by ultrasound-guided microwave ablation (MWA) for patients with large benign thyroid nodules (BTNs). METHODS: Eighty-seven patients with 87 BTNs (≥ 4 cm) treated with MWA between April 2015 and March 2021 were enrolled in this retrospective multicenter study, with clinical and ultrasound examinations performed at the 1st, 3rd, 6th, and 12th months. A multivariable linear mixed effects model was employed to explore the alterations in volume and volume reduction ratio (VRR), as well as the potential factors associated with VRR. RESULTS: The mean age of the 87 patients was 45.69 ± 14.21 years (range 18-76 years), and the ratio of men to women was 1:4.8. The mean volumes were much decreased at the 12th month after ablation compared to the initial volumes (p < .001). The mean VRR was 76.09% at the 12th month. The technique efficacy (VRR > 50%) was 90.80% at the 12th month. A multivariate analysis revealed that VRR was related to the initial volume (p = .015), annular flow (p = .010), and nodule composition (p = .024). The mean symptomatic score decreased from 4.40 ± 0.28 to 0.26 ± 0.06 at the 12th month (p < .001). At the same time, the mean cosmetic score decreased from 3.22 ± 0.10 to 1.31 ± 0.08 (p < .001). CONCLUSION: MWA could serve as a safe and effective therapy for large BTNs, significantly reducing the volume of BTNs and significantly improving compressive symptoms and appearance problems. CLINICAL RELEVANCE STATEMENT: Microwave ablation could serve as a safe and effective therapy for large benign thyroid nodules, leading to significant volume reduction and satisfied symptom and cosmetic alleviation period. KEY POINTS: • This multicenter study investigated the feasibility and safety of microwave ablation for large benign thyroid nodules. • After ablation, the nodule volume was significantly reduced, and patients' symptoms and appearance problems were significantly improved. • Microwave ablation is feasible for large benign thyroid nodules and has been a supplement treatment.

Access to Functionalized Cyclohex-2-enones from a Multicomponent Cascade Reaction of Readily Available Alkynes, Ketones, and Ethyl Acetoacetate.

The synthesis of cyclohex-2-enone derivatives is a topic of current interest in organic chemistry. A novel three-component cascade reaction of alkynes with ketones and ethyl acetoacetate has been uncovered. This process provides di- and trisubstituted cyclohex-2-enones in good yields with excellent functional group tolerance. A variety of terminal alkynes and a wide range of aryl, alkyl, and cyclic ketones are viable in this transformation. Successful scale-up preparation and synthetic transformations have demonstrated the potential of this simple operating protocol.

MMDTA: A Multimodal Deep Model for Drug-Target Affinity with a Hybrid Fusion Strategy.

The prediction of the drug-target affinity (DTA) plays an important role in evaluating molecular druggability. Although deep learning-based models for DTA prediction have been extensively attempted, there are rare reports on multimodal models that leverage various fusion strategies to exploit heterogeneous information from multiple different modalities of drugs and targets. In this study, we proposed a multimodal deep model named MMDTA, which integrated the heterogeneous information from various modalities of drugs and targets using a hybrid fusion strategy to enhance DTA prediction. To achieve this, MMDTA first employed convolutional neural networks (CNNs) and graph convolutional networks (GCNs) to extract diverse heterogeneous information from the sequences and structures of drugs and targets. It then utilized a hybrid fusion strategy to combine and complement the extracted heterogeneous information, resulting in the fused modal information for predicting drug-target affinity through the fully connected (FC) layers. Experimental results demonstrated that MMDTA outperformed the competitive state-of-the-art deep learning models on the widely used benchmark data sets, particularly with a significantly improved key evaluation metric, Root Mean Square Error (RMSE). Furthermore, MMDTA exhibited excellent generalization and practical application performance on multiple different data sets. These findings highlighted MMDTA's accuracy and reliability in predicting the drug-target binding affinity. For researchers interested in the source data and code, they are accessible at http://github.com/dldxzx/MMDTA.

Sleep disturbance and internalizing symptoms in adolescents: a moderated mediation model of self-control and mindfulness.

OBJECTIVES: Despite accumulating evidence regarding the impact of sleep disturbance on internalizing symptoms among adolescents, the underlying psychological mechanisms remain inadequately explored. This study aimed to investigate a conceptual framework elucidating how sleep disturbance influences internalizing symptoms in adolescents through the mediating role of self-control, with mindfulness as a moderator. METHODS: In this cross-sectional study, 1876 Chinese adolescents (Mage = 14.88 years, SD = 1.47 years, range = 12-19 years, 44.7% boys) completed the Youth Self-Rating Insomnia Scale (YSIS), Patient Health Questionnaire-9 (PHQ-9), Generalized Anxiety Disorder-7 (GAD-7), Brief Self-control Scale (BSCS), and Mindful Attention Awareness Scale-Children (MAAS-C) to provide data on sleep-related variables, internalizing symptoms (anxiety and depression), self-control, and mindfulness, respectively. The PROCESS macro for SPSS was applied to perform moderated mediation analysis. RESULTS: Sleep disturbance demonstrated a significant positive correlation with internalizing symptoms in adolescents, including anxiety (ß = 0.481, p < 0.001) and depression (ß = 0.543, p < 0.001). Self-control served as a mediator between sleep disturbance and two forms of internalizing symptoms. Moreover, mindfulness moderated the pathways from self-control to internalizing symptoms (for anxiety symptoms: ß = 0.007, p < 0.001; for depression symptoms: ß = 0.006, p < 0.001), and the mediating relationships were weaker for adolescents exhibiting higher levels of mindfulness. CONCLUSIONS: Our findings enhance understanding of the impact, pathways, and influencing factors of sleep disturbance on adolescent internalizing symptoms, suggesting the importance of enhancing mindfulness levels in addressing self-control deficits and subsequently reducing internalizing symptoms among adolescents.

Bidirectional associations between short sleep duration, insomnia symptoms, and psychotic-like experiences in adolescents.

This study investigates the prospective associations between short sleep duration, insomnia symptoms, and psychotic-like experiences (PLEs) in a large sample of Chinese adolescents. This study utilized a three-timepoint repeated cross-sectional survey with two nested longitudinal subsamples. A total of 17,722 adolescents were assessed at baseline (April 21 to May 12, 2021) and six months later (December 17 to 26, 2021). Out of these, 15,694 adolescents provided complete responses to the questions at baseline and one year later (May 17 - June 6, 2022). A self-administered questionnaire was used to measure sample characteristics (at baseline), sleep duration, insomnia symptoms, and PLEs (at each assessment), and negative life events (at two follow-ups). Baseline short sleep duration and insomnia symptoms predicted frequent PLEs at both follow-up assessments. Additionally, baseline frequent PLEs also predicted insomnia symptoms at six months and one year later. However, when controlling for confounders, PLEs at baseline only predicted short sleep duration at six months, and not at one year. This study reveals bidirectional prospective relationships between short sleep duration, insomnia symptoms, and PLEs, even after controlling for covariates. Therefore, it is crucial to assess both sleep patterns and PLEs in order to promote optimal sleep and mental health among adolescents.

Concurrent and longitudinal associations of dissatisfaction with height and weight, body mass index with sleep disturbance among adolescents in China.

Sleep disturbance in adolescents is a significant global public health issue that can result in various physical and mental disorders. Height and weight dissatisfaction, as subjective perceptions of body shape, are common in adolescence and may affect individuals' sleep situations. However, the association of them is unclear. This study aimed to examine the cross-sectional and longitudinal associations among height dissatisfaction, weight dissatisfaction, body mass index (BMI), and sleep disturbance in adolescents. A total of 27,260 participants completed measures of sleep disturbance, height and weight satisfaction, and BMI. The cross-sectional and longitudinal effects of height satisfaction, weight satisfaction, and BMI as well as their changes in sleep disturbance over time were tested. Height satisfaction (ps < 0.001) and weight satisfaction (ps < 0.001) were negatively associated with concurrent sleep disturbance after adjusting for socio-demographic covariates and previous sleep disturbance. Height satisfaction (ps < 0.001) and weight satisfaction (ps < 0.001) at T1 as well as their increases (ps < 0.001) over 6 months predicted decreases in sleep disturbance over time. The cross-sectional and longitudinal associations between BMI and sleep disturbance were not significant (ps > 0.42). Moreover, height dissatisfaction (ORs = 1.06 to 1.34) and weight dissatisfaction (ORs = 1.21 to 1.36) were risk factors for concurrent and long-term sleep disturbance while adjusting socio-demographic covariates and previous sleep disturbance. Greater attention to subjective perception of height satisfaction and weight satisfaction, instead of BMI is needed.

Mouse-INtraDuctal (MIND): an in vivo model for studying the underlying mechanisms of DCIS malignancy.

Due to widespread adoption of screening mammography, there has been a significant increase in new diagnoses of ductal carcinoma in situ (DCIS). However, DCIS prognosis remains unclear. To address this gap, we developed an in vivo model, Mouse-INtraDuctal (MIND), in which patient-derived DCIS epithelial cells are injected intraductally and allowed to progress naturally in mice. Similar to human DCIS, the cancer cells formed in situ lesions inside the mouse mammary ducts and mimicked all histologic subtypes including micropapillary, papillary, cribriform, solid, and comedo. Among 37 patient samples injected into 202 xenografts, at median duration of 9 months, 20 samples (54%) injected into 95 xenografts showed in vivo invasive progression, while 17 (46%) samples injected into 107 xenografts remained non-invasive. Among the 20 samples that showed invasive progression, nine samples injected into 54 xenografts exhibited a mixed pattern in which some xenografts showed invasive progression while others remained non-invasive. Among the clinically relevant biomarkers, only elevated progesterone receptor expression in patient DCIS and the extent of in vivo growth in xenografts predicted an invasive outcome. The Tempus XT assay was used on 16 patient DCIS formalin-fixed, paraffin-embedded sections including eight DCISs that showed invasive progression, five DCISs that remained non-invasive, and three DCISs that showed a mixed pattern in the xenografts. Analysis of the frequency of cancer-related pathogenic mutations among the groups showed no significant differences (KW: p > 0.05). There were also no differences in the frequency of high, moderate, or low severity mutations (KW; p > 0.05). These results suggest that genetic changes in the DCIS are not the primary driver for the development of invasive disease. © 2021 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Effect of mechanical ventilation under intubation on respiratory tract change of bacterial count and alteration of bacterial flora.

Background: The most common 'second strike' in mechanically ventilated patients is a pulmonary infection caused by the ease with which bacteria can invade and colonize the lungs due to mechanical ventilation. At the same time, metastasis of lower airway microbiota may have significant implications in developing intubation mechanical ventilation lung inflammation. Thus, we establish a rat model of tracheal intubation with mechanical ventilation and explore the effects of mechanical ventilation on lung injury and microbiological changes in rats. To provide a reference for preventing and treating bacterial flora imbalance and pulmonary infection injury caused by mechanical ventilation of tracheal intubation. Methods: Sprague-Dawley rats were randomly divided into Control, Mechanical ventilation under intubation (1, 3, 6 h) groups, and Spontaneously breathing under intubation (1, 3, 6 h). Lung histopathological injury scores were evaluated. 16SrDNA sequencing was performed to explore respiratory microbiota changes, especially, changes of bacterial count and alteration of bacterial flora. Results: Compared to groups C and SV, critical pathological changes in pulmonary lesions occurred in the MV group after 6 h (p < 0.05). The Alpha diversity and Beta diversity of lower respiratory tract microbiota in MV6, SV6, and C groups were statistically significant (p < 0.05). The main dominant bacterial phyla in the respiratory tract of rats were Proteobacteria, Firmicutes, Bacteroidetes, and Cyanobacteria. Acinetobacter radioresistens in group C was significant, Megaonas in group MV6 was significantly increased, and Parvibacter in group SV6 was significantly increased. Anaerobic, biofilm formation, and Gram-negative bacteria-related functional genes were altered during mechanical ventilation with endotracheal intubation. Conclusion: Mechanical ventilation under intubation may cause dysregulation of lower respiratory microbiota in rats.

Trajectory of adolescents' nightmares in relation to depression and post-traumatic stress disorder in young adulthood: a 10-year cohort of Chinese Wenchuan earthquake survivors.

PURPOSE: This study examined the longitudinal trajectories of adolescent survivors' nightmares after the Wenchuan earthquake and tested whether specific trajectory memberships of nightmares could predict depression and post-traumatic stress disorder (PTSD) in the 10 years after earthquake. METHODS: 610 adolescents exposed to the Wenchuan earthquake were surveyed on nightmares at 18 months (T18m), 24 months (T24m), and 30 months (T30m) after the earthquake. Depression and PTSD were assessed at baseline (T18m) and 10 years (T10y) after the catastrophe. Data were analyzed using linear regression analysis. RESULTS: The prevalence rates of frequent nightmares at three time points were 9.8%, 11.5%, and 9.3%, respectively. Five different trajectories of nightmares were identified: resistance (77.0%), recover (8.4%), delayed-dysfunction (7.9%), chronic-dysfunction (1.1%), and relapsing/remitting (5.6%). Additionally, we found that participants in relapsing/remitting, chronic or delayed dysfunction trajectories compared with those in the resistant group were more likely to experience depression and PTSD in young adulthood after adjusting for a wide range of covariates. CONCLUSIONS: Nightmares had heterogeneity after a catastrophic earthquake. Timely assessment and targeted interventions on specific nightmares are necessary for reducing the incidence of psychiatric disorders.

Longitudinal associations between social support trajectories and quality of life mediated by depressive symptoms: A 10-year cohort study of Wenchuan earthquake.

INTRODUCTION: Previous studies on associations between social support and quality of life (QoL) have usually employed a variable-centered approach, without considering individual variances in time-varying changes in social support. This study explores trajectories of social support and whether depressive symptoms mediate associations between social support trajectories and QoL 10 years after an earthquake. METHODS: Seven hundred and forty-four Chinese adolescents exposed to the Wenchuan earthquake were surveyed on social support at 6-, 18-, and 24-months and depressive symptoms at 30-months postearthquake (T30m ). They provided valid data on QoL after 10 years of the earthquake (T10y ). The latent class growth analysis was used to estimate social support trajectories. Mediation analysis was then conducted to test whether depressive symptoms at T30m mediated associations between social support trajectories and QoL at T10y . RESULTS: Three trajectories of social support were identified: low decreasing (31.6%), moderate decreasing (55.4%), and persistent high (13.0%) groups. Depressive symptoms significantly mediated the effects of social support trajectories (relative to the low support trajectory) on future QoL (95% CIs: 0.70-1.78 and 1.41-3.37 for moderate decreasing and persistent high groups, respectively). CONCLUSIONS: Social support shows individual differences over time. Moderate and high social support trajectories improve 10-year QoL partly by reducing depressive symptoms. Therefore, interventions aimed at enhancing social support and reducing depressive symptoms may be more effective in enhancing QoL in the aftermath of disasters.

Network Analysis of Health-related Behaviors, Insomnia, and Depression Among Urban Left-behind Adolescents in China.

Mental health of urban left-behind adolescents (LBA) is a public issue of growing concern. This study aims to examine the symptom level associations among multiple health-related behaviors, insomnia, and depression in urban LBA. Data on a sample of urban LBA aged 11-19 (N = 3,601) from the Adolescent Mental Health Survey in Shenzhen, China, were used. Health-related behaviors (i.e., Internet use, physical inactivity, social jetlag, smoking, and alcohol consumption), insomnia, and depressive symptoms were assessed using a self-administered questionnaire. Graphical Gaussian Model (GGM) was used to describe key bridging nodes in an undirected network. Directed Acyclic Graph (DAG) was used to construct a directed network and estimate the most likely causal associations among behaviors/symptoms. In the undirected network, Internet use was identified as the key bridging node most strongly associated with insomnia and depression. Two other key bridging nodes include difficulty initiating sleep and appetite change. In the directed network, anhedonia emerged as the most pivotal symptom, which could cause insomnia symptoms and behavioral changes, either directly, or through triggering other depressive symptoms, such as low energy and appetite change. These findings have implications for understanding the occurrence and maintenance process of health-related behaviors, insomnia, and depression in urban LBA. In practice, Internet use should be considered a priority in targeting multiple health behavior interventions. Meanwhile, early screening and treatment for anhedonia are of great significance as well.

The More, the Better? Social Capital Profiles and Adolescent Internalizing Symptoms: A Latent Profile Analysis.

Past research suggests that offline and online social capital are empirically linked to adolescent psychological adjustment. However, little is known regarding the implications of distinctive combinations of social capital for adolescent internalizing symptoms. The present study aimed to examine adolescent social capital patterns and their associations with internalizing symptoms by using latent profile analysis. A cross-sectional web-based survey was conducted among 1595 Chinese adolescents (mean age = 14.30 years, 50.7% male). All adolescents completed self-report questionnaires on their perceived offline and online social capital, depressive symptoms and anxiety symptoms. Latent profile analysis revealed four profiles of social capital: (1) Low Social Capital, (2) Moderate Social Capital, (3) High Social Capital, and (4) Only High Offline Social Capital. Further, analysis of covariance demonstrated that the Only High Offline Social Capital profile had significantly fewer internalizing symptoms than other three profiles. No statistical differences of internalizing symptoms were found between the other three profiles, except that the Moderate Social Capital profile showed fewer anxiety symptoms than the Low Social Capital profile. These findings suggest that more social capital does not equal to better mental health status. The social capital profiles and their associations with adolescent internalizing symptoms may provide practitioners with meaningful implications regarding the role of offline and online social capital in adolescent psychological adjustment.

Associations Between Family Function and Non-suicidal Self-injury Among Chinese Urban Adolescents with and Without Parental Migration.

The study aims to explore the effect of family function on non-suicidal self-injury (NSSI) among Chinese urban adolescents with and without parental migration. Between April 21st to May 12th, 2021, adolescents were recruited from Shenzhen city of Guangdong province, China (n = 124,357). Of all the participants, 22,855 (18.4%) were left-behind children (LBC). Family function, NSSI, depression, and socio-demographic characteristics were assessed using a series of self-reported questionnaires. Urban LBC had a higher NSSI frequency, while a lower level of family function than non-LBC. After controlling for confounders, parental migration was significantly associated with NSSI, and family dysfunction was a robust risk factor for NSSI as well. The protective effect of family function on NSSI of LBC was stronger than non-LBC. This implies that children with higher levels of family function tend to exhibit a lower frequency of NSSI, especially in those with parental migration. In practice, adolescents' NSSI prevention and intervention strategies should focus on improving family function.

Profiles of Intolerance of Uncertainty Among 108,540 Adolescents: Associations with Sociodemographic Variables and Mental Health.

Intolerance of uncertainty (IU) is widely considered a transdiagnostic risk and maintaining factor for psychiatric disorders. However, little is known about the overall nature and profile of IU among adolescents. This study aims to investigate the profiles of IU among Chinese adolescents and explore their associations with sociodemographic characteristics and mental health problems. A sample of 108,540 adolescents provided data on IU, sociodemographic characteristics, and mental health via an online platform. Latent profile analysis revealed three profiles: Low IU, Medium IU, and High IU. Girls, older adolescents, and those with specific sociodemographics were more likely to belong to the "High IU" profile. Furthermore, the "High IU" profile was associated with the highest risk of several mental health problems. These findings provided valuable information for early prevention and intervention strategies targeting IU and highlighted the importance of IU-based interventions for mental health among adolescents.

[Related factors of endogenous erythropoietin and its association with 10-year risks of cardiovascular disease in a community-based Chinese study].

OBJECTIVE: To investigate the associated factors of endogenous erythropoietin (EPO) and its association with 10-year risks of atherosclerotic cardiovascular disease in a Chinese community-based general population. METHODS: The participants of this study were from an atherosclerosis cohort survey which was established by the Department of Cardiology, Peking University First Hospital in 2011. The cohort survey was performed in the Gucheng and Pingguoyuan communities of Shijingshan district in Beijing, China. The inclusion criteria of this study were: (1) endogenous EPO was measured; (2) health questionnaire data and other clinical data were complete; (3) participatants who had cardiovascular or cerebrovascular diseases (defined as self-reported coronary heart disease, stroke or transient ischemic attack) or anemia or estimated glomerular filtration rate (eGFR) < 60 mL/(min·1.73 m2) at baseline were excluded. Multivariate linear regression model was used to examine the associated factors of endogenous EPO. The participants were grouped into low (< 5%), moderate (5%-10%) and high risk (≥10%) groups, based on predicted 10-year cardiovascular disease risk using the prediction for atherosclerotic cardiovascular disease risk in China (China-PAR) equations. RESULTS: A total of 4 013 participants were included. Mean age of them was (55.9±8.2) years, 62.2% (n=2 496) of them were female, and 46.3% (n=1 859), 70.9% (n=2 845), 21.9% (n=879) had hypertension, dyslipidemia and diabetes, individually. The average body mass index was (26.1±3.3) kg/m2. The median of EPO level was 12.8 (9.3-17.4) IU/L and 25.1% (n=998) were at high 10-years risk of cardiovascular disease. Hemoglobin (ß=-0.05, 95%CI: -0.07 to -0.04) and eGFR ≥90 mL/(min·1.73 m2) (ß=-0.05, 95%CI: -0.07 to -0.04) were associated with lower in transformed EPO levels while hypertension (ß=0.08, 95%CI: 0.05 to 0.12) and obesity (ß=0.14, 95%CI: 0.09 to 0.18) were associated with higher in transformed EPO levels in multivariate linear regression analyses. Ten-year cardiovascular disease risks were positively associated with in transformed EPO levels (ß=0.07, 95%CI: 0.05 to 0.09). The participants at moderate and high cardiovascular disease risks had significant higher EPO levels than the low risk group (all P < 0.05). CONCLUSION: In community-based Beijing populations, endogenous EPO was associated with hemoglobin, renal function, obesity and hypertension. Individuals at high 10-years cardiovascular disease risks have higher endogenous EPO levels. Endogenous EPO may be a potential risk marker of cardiovascular disease.

[Extraction of Extracelluar Vesicles Derived from Mycobacterium tuberculosis and Their Effect on the Production of Reactive Oxygen Species and Expression of Inflammatory Factors in Mouse Bone Marrow-Derived Dendritic Cells].

Objective: To isolate extracellular vesicles (EVs) from Mycobacterium tuberculosis ( Mtb), to examine their morphology, particle size, and distribution, to study the effect of EVs derived from Mtb ( Mtb-EVs) on intracellular reactive oxygen species (ROS) production and cytokine secretion in dendritic cells (DCs), and to make preliminary exploration of Mtb-EVs' effect on the immune regulation of DCs. Methods: Mtb-EVs were obtained by ultrafiltration concentration and the protein concentration was determined by BCA assay. The morphology of Mtb-EVs was observed through negative staining electron microscopy (EM). The particle size distribution and concentration of Mtb-EVs were determined by nanoparticle tracking analysis (NTA). Mouse bone marrow was isolated through sterile procedures and mice myeloid DCs were induced and amplified by the combined use of recombinant mouse granulocyte-macrophage colony-stimulating factor (rm GM-CSF) and recombinant mouse interleukin-4 (rm IL-4). Then, morphological and immunophenotypic characterization was performed. After that, the DCs were treated with Mtb-EVs at different concentrations and CCK-8 assay was done to measure their effect on the survival rate of DCs and to identify the appropriate stimulation concentration for subsequent experimental procedures. The intracellular ROS levels of DCs were evaluated with DCFH-DA fluorescence probe and the cytokine secretion of DCs was determined by ELISA. Results: EM observation showed that Mtb-EVs isolated by ultrafiltration concentration were spherical vesicles of varied sizes, all being approximately 100 nm in diameter and displaying typical morphology. NTA results from NanoSight nanoparticle tracker showed that the peak particle size was 98.5 nm, that the average particle size was 110.2 nm, and that the particle size was mainly distributed between 68.4-155.7 nm. Mtb-EVs that were smaller than 250 nm accounted for 98.39% of the total. Mouse myeloid DCs directionally induced and amplified in vitro displayed typical DC phenotype and morphology, and the purity exceeded 85%. EM verified the abundance of microvilli and radial protuberance on the surface of DCs, which had uniform cytoplasm and clear nuclear membrane. Loaded with Mtb-EVs at different concentrations, including 10 2, 10 3, and 10 4 particles/cell, the DCs had significantly upregulated levels of intracellular ROS ( P<0.05). In addition, Mtb-EVs induced the release of IL-1ß and IL-6 in a dose-dependent manner ( P<0.05). Conclusion: We established in the study a technical process for the extraction of Mtb-EVs by ultrafiltration concentration and obtained Mtb-EVs with sound morphology, high purity, and concentrated particle size distribution. Furthermore, Mtb-EVs can upregulate the intracellular ROS level in DCs and induce the release of IL-1ß and IL-6 in a dose-dependent manner.

Residual cancer burden after neoadjuvant chemotherapy and long-term survival outcomes in breast cancer: a multicentre pooled analysis of 5161 patients.

BACKGROUND: Previous studies have independently validated the prognostic relevance of residual cancer burden (RCB) after neoadjuvant chemotherapy. We used results from several independent cohorts in a pooled patient-level analysis to evaluate the relationship of RCB with long-term prognosis across different phenotypic subtypes of breast cancer, to assess generalisability in a broad range of practice settings. METHODS: In this pooled analysis, 12 institutes and trials in Europe and the USA were identified by personal communications with site investigators. We obtained participant-level RCB results, and data on clinical and pathological stage, tumour subtype and grade, and treatment and follow-up in November, 2019, from patients (aged ≥18 years) with primary stage I-III breast cancer treated with neoadjuvant chemotherapy followed by surgery. We assessed the association between the continuous RCB score and the primary study outcome, event-free survival, using mixed-effects Cox models with the incorporation of random RCB and cohort effects to account for between-study heterogeneity, and stratification to account for differences in baseline hazard across cancer subtypes defined by hormone receptor status and HER2 status. The association was further evaluated within each breast cancer subtype in multivariable analyses incorporating random RCB and cohort effects and adjustments for age and pretreatment clinical T category, nodal status, and tumour grade. Kaplan-Meier estimates of event-free survival at 3, 5, and 10 years were computed for each RCB class within each subtype. FINDINGS: We analysed participant-level data from 5161 patients treated with neoadjuvant chemotherapy between Sept 12, 1994, and Feb 11, 2019. Median age was 49 years (IQR 20-80). 1164 event-free survival events occurred during follow-up (median follow-up 56 months [IQR 0-186]). RCB score was prognostic within each breast cancer subtype, with higher RCB score significantly associated with worse event-free survival. The univariable hazard ratio (HR) associated with one unit increase in RCB ranged from 1·55 (95% CI 1·41-1·71) for hormone receptor-positive, HER2-negative patients to 2·16 (1·79-2·61) for the hormone receptor-negative, HER2-positive group (with or without HER2-targeted therapy; p<0·0001 for all subtypes). RCB score remained prognostic for event-free survival in multivariable models adjusted for age, grade, T category, and nodal status at baseline: the adjusted HR ranged from 1·52 (1·36-1·69) in the hormone receptor-positive, HER2-negative group to 2·09 (1·73-2·53) in the hormone receptor-negative, HER2-positive group (p<0·0001 for all subtypes). INTERPRETATION: RCB score and class were independently prognostic in all subtypes of breast cancer, and generalisable to multiple practice settings. Although variability in hormone receptor subtype definitions and treatment across patients are likely to affect prognostic performance, the association we observed between RCB and a patient's residual risk suggests that prospective evaluation of RCB could be considered to become part of standard pathology reporting after neoadjuvant therapy. FUNDING: National Cancer Institute at the US National Institutes of Health.

Hydroamination of Unactivated Alkenes with Aliphatic Azides.

We report here an efficient and highly diastereoselective intermolecular anti-Markovnikov hydroamination of unactivated alkenes with aliphatic azides in the presence of silane. The system tolerates a wide range of azides and alkenes and operates with alkene as limiting reagent. Mechanistic studies suggest a radical chain pathway that involves aminium radical formation, radical addition to alkenes and HAT from silane to ß-aminium alkyl radical. The use of sterically bulky silane is proposed to contribute to the excellent diastereoselectivity for HAT. Computational analysis uncovers the reaction pathway of aliphatic azide activation with silyl radical for aminyl radical formation.