Bioprinted, spatially defined breast tumor microenvironment models of intratumoral heterogeneity and drug resistance.

Cellular, extracellular matrix (ECM), and spatial heterogeneity of tumor microenvironments (TMEs) regulate disease progression and treatment efficacy. Developing in vitro models that recapitulate the TME promises to accelerate studies of tumor biology and identify new targets for therapy. Here, we used extrusion-based, multi-nozzle 3D bioprinting to spatially pattern triple-negative MDA-MB-231 breast cancer cells, endothelial cells (ECs), and human mammary cancer-associated fibroblasts (HMCAFs) with biomimetic ECM inks. Bioprinted models captured key features of the spatial architecture of human breast tumors, including varying-sized dense regions of cancer cells and surrounding microvessel-rich stroma. Angiogenesis and ECM stiffening occurred in the stromal area but not the cancer cell-rich (CCR) regions, mimicking pathological changes in patient samples. Transcriptomic analyses revealed upregulation of angiogenesis-related and ECM remodeling-related signatures in the stroma region and identified potential ligand-receptor (LR) mediators of these processes. Breast cancer cells in distinct parts of the bioprinted TME showed differing sensitivities to chemotherapy, highlighting environmentally mediated drug resistance. In summary, our 3D-bioprinted tumor model will act as a platform to discover integrated functions of the TME in cancer biology and therapy.

Optimal scheduling model using the IGDT method for park integrated energy systems considering P2G-CCS and cloud energy storage.

To enhance the energy efficiency and financial gains of the park integrated energy system (PIES). This paper constructs a bi-level optimization model of PIES-cloud energy storage (CES) based on source-load uncertainty. Firstly, the scheduling framework of PIES with refined power-to-gas (P2G), carbon capture and storage (CCS) and CES coupling is constructed. Moreover, a bi-level optimization model with the upper tier subject being the PIES operator and the lower tier subject being the CES operator is established under the ladder-type carbon price mechanism with reward and punishment (LCPMRP). Then a proposed entropy weight adaptive information gap decision theory method (EAIGDT) is proposed to eliminate the subjectivity factor and retain its non-probabilistic features while dealing with multiple source-load uncertainties, and according to the operator's risk preference to build risk-averse (RA) and risk-seeking (RS) strategies, respectively. Finally, the measured data in a certain area of Xinjiang verifies the proposed optimal scheduling method. The results show that the method can effectively take into account the interests of various subjects and realise PIES low-carbon economic operation.

Machine learning-based prediction of diabetic patients using blood routine data.

Diabetes stands as one of the most prevalent chronic diseases globally. The conventional methods for diagnosing diabetes are frequently overlooked until individuals manifest noticeable symptoms of the condition. This study aimed to address this gap by collecting comprehensive datasets, including 1000 instances of blood routine data from diabetes patients and an equivalent dataset from healthy individuals. To differentiate diabetes patients from their healthy counterparts, a computational framework was established, encompassing eXtreme Gradient Boosting (XGBoost), random forest, support vector machine, and elastic net algorithms. Notably, the XGBoost model emerged as the most effective, exhibiting superior predictive results with an area under the receiver operating characteristic curve (AUC) of 99.90% in the training set and 98.51% in the testing set. Moreover, the model showcased commendable performance during external validation, achieving an overall accuracy of 81.54%. The probability generated by the model serves as a risk score for diabetes susceptibility. Further interpretability was achieved through the utilization of the Shapley additive explanations (SHAP) algorithm, identifying pivotal indicators such as mean corpuscular hemoglobin concentration (MCHC), lymphocyte ratio (LY%), standard deviation of red blood cell distribution width (RDW-SD), and mean corpuscular hemoglobin (MCH). This enhances our understanding of the predictive mechanisms underlying diabetes. To facilitate the application in clinical and real-life settings, a nomogram was created based on the logistic regression algorithm, which can provide a preliminary assessment of the likelihood of an individual having diabetes. Overall, this research contributes valuable insights into the predictive modeling of diabetes, offering potential applications in clinical practice for more effective and timely diagnoses.

Integrated analysis of patients with bladder cancer from prospective transcription factor activity: Implications for personalized treatment approaches.

Transcription factors are a specialized group of proteins that play important roles in regulating gene expression in human. These proteins control the transcription and translation of genes by binding to specific sites on DNA, thereby regulating key biological processes such as cell differentiation, proliferation, immune response, and neural development. Moreover, transcription factors are also involved in apoptosis and the pathogenesis of various diseases. By investigating transcription factors, researchers can uncover the mechanisms of gene regulation in organisms and develop more effective methods for preventing and treating human diseases. In the present study, the Virtual Inference of Protein-activity by Enriched Regulon algorithm was utilized to calculate the protein activity of transcription factors, and the metabolic-related protein activity were used for classifying bladder cancer patients into different subtype. To identify chemotherapy drugs with clinical benefits, the differences in prognosis and drug sensitivity between two distinct subtypes of bladder cancer patients were investigated. Simultaneously, the master regulators that display varying levels of transcription factor activity between two different bladder cancer subtypes were explored. Additionally, the potential transcriptional regulatory mechanisms and targets of these factors were investigated, thereby generating novel insights into bladder cancer research at the transcriptional regulation level.

The effect of a hybrid structured pulmonary rehabilitation education program for patients with lung cancer with a high risk of postoperative pulmonary complications: A quasi-experimental study.

PURPOSE: The absence of standardized protocols and education are the main obstacles to perioperative pulmonary rehabilitation (PR), especially for patients with high-risk factors of postoperative pulmonary complications (PPCs). We aimed to explore the effect of a hybrid structured pulmonary rehabilitation education program (SPREP) on patients with lung cancer at high risk of PPCs. METHODS: A quasi-experimental trial with a pre-post test design was conducted. The control group (n = 53) adopted routine perioperative pulmonary rehabilitation, while the intervention group (n = 53) received SPREP. Respiratory function, 6-min walk distance, Borg dyspnea scale, quality of life, anxiety-depression scores at admission, discharge, 2 weeks and 3 months post-discharge, and incidence of PPCs were compared between the two groups. RESULTS: There were no significant differences on the 6-min walk distance and Borg Dyspnoea Scale at discharge between the two groups (P > 0.05), whereas the intervention group showed improved performance at the remaining time points (P < 0.05). In addition, the intervention group had improved exercise capacity, pulmonary function and quality of life, reduced levels of anxiety and depression at discharge, 2 weeks post-discharge and 3 months post-discharge (P < 0.05). In addition, incidence of PPCs was significantly reduced in the intervention group, especially postoperative pneumonia. CONCLUSIONS: The SPREP could show significant benefits in enhancing exercise capacity, lung function, and quality of life, while diminishing the occurrence of PPCs and mitigating the levels of anxiety and depression, future large RCT need to further explore the efficacy. TRIAL REGISTRATION: This study was registered with the China Clinical Trial Registration Center (ChiCTR) under the Clinical Trial Registration Number [ChiCTR2200066698].

Hybridization of short-range and long-range charge transfer boosts room-temperature phosphorescence performance.

At present, mainstream room-temperature phosphorescence (RTP) emission relies on organic materials with long-range charge-transfer effects; therefore, exploring new forms of charge transfer to generate RTP is worth studying. In this work, indole-carbazole was used as the core to ensure the narrowband fluorescence emission of the material based on its characteristic short-range charge-transfer effect. In addition, halogenated carbazoles were introduced into the periphery to construct long-range charge transfer, resulting in VTCzNL-Cl and VTCzNL-Br. By encapsulating these phosphors into a robust host (TPP), two host-guest crystalline systems were further developed, achieving efficient RTP performance with phosphorescence quantum yields of 26% and phosphorescence lifetimes of 3.2 and 39.2 ms, respectively.

Impact of prior SARS-CoV-2 infection on college students' hesitancy to receive additional COVID-19 vaccine booster doses: A study from Taizhou, China.

Purpose: This study aimed to examine the impact of a history of SARS-CoV-2 infection on the hesitancy of college students to receive additional COVID-19 vaccine booster doses. Methods: A population-based self-administered online survey was conducted in July 2024 in Taizhou, China. A total of 792 respondents were included in this study. Logistic regression was conducted to identify factors associated with college students' hesitation to receive booster doses of the COVID-19 vaccine. Results: Of 792 respondents, 32.2 % hesitated to receive additional doses of the COVID-19 vaccine booster. Furthermore, 23.5 % of the respondents reported an increase in hesitancy to receiving additional COVID-19 vaccine booster doses compared to before they were infected with SARS-CoV-2. In the regression analyses, college students who had a secondary infection were more hesitant to receive additional COVID-19 vaccine booster doses (OR = 0.481, 95 % CI: (0.299-0.774), P = 0.003). Moreover, students with secondary infections who were male (OR = 0.417, 95 % CI: 0.221-0.784, P = 0.007), with lower than a bachelor's degree (OR = 0.471, 95 % CI: 0.272-0.815, P = 0.007), in non-medical majors (OR = 0.460, 95 % CI: 0.248-0.856, P = 0.014), and sophomores or below (OR = 0.483, 95 % CI: 0.286-0.817, P = 0.007) were more hesitant to receive additional COVID-19 vaccine booster doses. Conclusion: A history of SARS-CoV-2 infection affects college students' hesitation to receive additional COVID-19 vaccine booster doses, which was higher in those who experienced secondary infections.

Implicit weight bias: shared neural substrates for overweight and angry facial expressions revealed by cross-adaptation.

The perception of facial expression plays a crucial role in social communication, and it is known to be influenced by various facial cues. Previous studies have reported both positive and negative biases toward overweight individuals. It is unclear whether facial cues, such as facial weight, bias facial expression perception. Combining psychophysics and event-related potential technology, the current study adopted a cross-adaptation paradigm to examine this issue. The psychophysical results of Experiments 1A and 1B revealed a bidirectional cross-adaptation effect between overweight and angry faces. Adapting to overweight faces decreased the likelihood of perceiving ambiguous emotional expressions as angry compared to adapting to normal-weight faces. Likewise, exposure to angry faces subsequently caused normal-weight faces to appear thinner. These findings were corroborated by bidirectional event-related potential results, showing that adaptation to overweight faces relative to normal-weight faces modulated the event-related potential responses of emotionally ambiguous facial expression (Experiment 2A); vice versa, adaptation to angry faces relative to neutral faces modulated the event-related potential responses of ambiguous faces in facial weight (Experiment 2B). Our study provides direct evidence associating overweight faces with facial expression, suggesting at least partly common neural substrates for the perception of overweight and angry faces.

[Cadmium Phytoremediation Effect of Sweet Sorghum Assisted with Citric Acid on Typical Parent Soil in Southern China].

Sweet sorghum has a large biomass and strong cadmium (Cd) absorption capacity, which has the potential for phytoremediation of Cd-contaminated soil. In order to study the Cd phytoremediation effect of sweet sorghum assisted with citric acid on the typical parent materials in southern China, a field experiment was carried out in two typical parent material farmland areas (neutral purple mud field and jute sand mud field) with Cd pollution in Hunan Province. The results showed that:① Citric acid had no inhibitory effect on the growth of sweet sorghum. After the application of citric acid, the aboveground biomass of sweet sorghum at the maturity stage increased by 10.1%-24.7%. ② Both sweet sorghum planting and citric acid application reduced the soil pH value, and the application of citric acid further reduced the soil pH value at each growth stage of sweet sorghum; this decrease was greater in the neutral purple mud field, which decreased by 0.24-0.72 units. ③ Both sweet sorghum planting and citric acid application reduced the total amount of soil Cd, and the decreases in the neutral purple mud field and jute sand mud field were 23.8%-52.2% and 17.1%-31.8%, respectively. The acid-extractable percentage of soil Cd in both places increased by 38.6%-147.7% and 4.8%-22.7%, respectively. ④ The application of citric acid could significantly increase the Cd content in various tissues of sweet sorghum. The Cd content in the aboveground part of the plant in the neutral purple mud field was higher than that in the jute sand mud field, and the Cd content in stems and leaves was 0.25-1.90 mg·kg-1 and 0.21-0.64 mg·kg-1, respectively. ⑤ After applying citric acid, the Cd extraction amount of sweet sorghum in neutral purple mud soil in the mature stage reached 47.56 g·hm-2. In summary, citric acid could enhance the efficiency of sweet sorghum in the phytoremediation of Cd-contaminated soil, and the effect was better in neutral purple mud fields. This technology has the potential for remediation coupled with agro-production for heavy metal-contaminated farmland.

Trophic Niche Differentiation in Two Sympatric Nuthatch Species (Sitta yunnanensis and Sitta nagaensis).

Sympatric closely related species may experience interspecific trophic competition due to ecological similarity; they may isolate in terms of diet or habitat use as a strategy to avoid competition. The body tissues of consumers contain stable isotope signatures information that can be applied to infer their dietary information. In this study, δ13C and δ15N stable isotopes were analyzed to determine the dietary information and trophic niches of sympatric coexisting Sitta yunnanensis and Sitta nagaensis. The results showed that the food sources of S. yunnanensis and S. nagaensis were from six orders, including Orthoptera, and the cumulative contribution rate was 99.97%, with the two species eating similar diets but at different rates. The larger δ13C of S. yunnanensis indicates that it had a wider range of habitats for feeding, while the difference in δ15N values was not significant (p > 0.05), indicating that both species feed on similar nutrient levels. As determined by Bayesian ellipses, the isotopic niches of S. yunnanensis and S. nagaensis were differentiated; the isotopic niche width of S. yunnanensis is 2.69‱2, which was larger than that of S. nagaensis (0.73‱2), indicates that differentiation between the two species in diet or habitat use reduced competition. Trophic niche differentiation and differences in foraging proportions may be the principal resource allocation mechanisms behind S. yunnanensis and S. nagaensis coexistence.

Machine learning applied to epilepsy: bibliometric and visual analysis from 2004 to 2023.

Background: Epilepsy is one of the most common serious chronic neurological disorders, which can have a serious negative impact on individuals, families and society, and even death. With the increasing application of machine learning techniques in medicine in recent years, the integration of machine learning with epilepsy has received close attention, and machine learning has the potential to provide reliable and optimal performance for clinical diagnosis, prediction, and precision medicine in epilepsy through the use of various types of mathematical algorithms, and promises to make better parallel advances. However, no bibliometric assessment has been conducted to evaluate the scientific progress in this area. Therefore, this study aims to visually analyze the trend of the current state of research related to the application of machine learning in epilepsy through bibliometrics and visualization. Methods: Relevant articles and reviews were searched for 2004-2023 using Web of Science Core Collection database, and bibliometric analyses and visualizations were performed in VOSviewer, CiteSpace, and Bibliometrix (R-Tool of R-Studio). Results: A total of 1,284 papers related to machine learning in epilepsy were retrieved from the Wo SCC database. The number of papers shows an increasing trend year by year. These papers were mainly from 1,957 organizations in 87 countries/regions, with the majority from the United States and China. The journal with the highest number of published papers is EPILEPSIA. Acharya, U. Rajendra (Ngee Ann Polytechnic, Singapore) is the authoritative author in the field and his paper "Deep Convolutional Neural Networks for Automated Detection and Diagnosis of Epileptic Seizures Using EEG Signals" was the most cited. Literature and keyword analysis shows that seizure prediction, epilepsy management and epilepsy neuroimaging are current research hotspots and developments. Conclusions: This study is the first to use bibliometric methods to visualize and analyze research in areas related to the application of machine learning in epilepsy, revealing research trends and frontiers in the field. This information will provide a useful reference for epilepsy researchers focusing on machine learning.

A bionic self-driven retinomorphic eye with ionogel photosynaptic retina.

Bioinspired bionic eyes should be self-driving, repairable and conformal to arbitrary geometries. Such eye would enable wide-field detection and efficient visual signal processing without requiring external energy, along with retinal transplantation by replacing dysfunctional photoreceptors with healthy ones for vision restoration. A variety of artificial eyes have been constructed with hemispherical silicon, perovskite and heterostructure photoreceptors, but creating zero-powered retinomorphic system with transplantable conformal features remains elusive. By combining neuromorphic principle with retinal and ionoelastomer engineering, we demonstrate a self-driven hemispherical retinomorphic eye with elastomeric retina made of ionogel heterojunction as photoreceptors. The receptor driven by photothermoelectric effect shows photoperception with broadband light detection (365 to 970 nm), wide field-of-view (180°) and photosynaptic (paired-pulse facilitation index, 153%) behaviors for biosimilar visual learning. The retinal photoreceptors are transplantable and conformal to any complex surface, enabling visual restoration for dynamic optical imaging and motion tracking.

The relationship between emotional disorders and heart rate variability: A Mendelian randomization study.

OBJECTIVE: Previous studies have shown that emotional disorders are negatively associated with heart rate variability (HRV), but the potential causal relationship between genetic susceptibility to emotional disorders and HRV remains unclear. We aimed to perform a Mendelian randomization (MR) study to investigate the potential association between emotional disorders and HRV. METHODS: The data used for this study were obtained from publicly available genome-wide association study datasets. Five models, including the inverse variance weighted model (IVW), the weighted median estimation model (WME), the weighted model-based method (WM), the simple model (SM) and the MR-Egger regression model (MER), were utilized for MR. The leave-one-out sensitivity test, MR pleiotropy residual sum and outlier test (MR-PRESSO) and Cochran's Q test were used to confirm heterogeneity and pleiotropy. RESULTS: MR analysis revealed that genetic susceptibility to broad depression was negatively correlated with HRV (pvRSA/HF) (OR = 0.380, 95% CI 0.146-0.992; p = 0.048). However, genetic susceptibility to irritability was positively correlated with HRV (pvRSA/HF, SDNN) (OR = 2.017, 95% CI 1.152-3.534, p = 0.008) (OR = 1.154, 95% CI 1.000-1.331, p = 0.044). Genetic susceptibility to anxiety was positively correlated with HRV (RMSSD) (OR = 2.106, 95% CI 1.032-4.299; p = 0.041). No significant directional pleiotropy or heterogeneity was detected. The accuracy and robustness of these findings were confirmed through a sensitivity analysis. CONCLUSIONS: Our MR study provides genetic support for the causal effects of broad depression, irritable mood, and anxiety on HRV.

Acid tolerance response of Salmonella during the squid storage and its amine production capacity analysis.

Salmonella exhibits a strong inducible acid tolerance response (ATR) under weak acid conditions, and can also induce high-risk strains that are highly toxic, acid resistant, and osmotic pressure resistant to aquatic products. However, the induction mechanism is not yet clear. Therefore, this study aims to simulate the slightly acidic, low-temperature, and high-protein environment during squid processing and storage. Through λRed gene knockout, exploring the effects of low-acid induction, long-term low-temperature storage, and two-component regulation on Salmonella ATR. In this study, we found the two-component system, PhoP/PhoQ and PmrA/PmrB in Salmonella regulates the amino acid metabolism system and improves bacterial acid tolerance by controlling arginine and lysine. Compared with the two indicators of total biogenic amine and diamine content, biogenic amine index and quality index were more suitable for evaluating the quality of aquatic products. The result showed that low-temperature treatment could inhibit Salmonella-induced ATR, which further explained the ATR mechanism from the amino acid metabolism.

Syringin alleviates bisphenol A-induced spermatogenic defects and testicular injury by suppressing oxidative stress and inflammation in male zebrafish.

Syringin (SRG) is a bioactive principle possessing extensive activities including scavenging of free radicals, inhibition of apoptosis, and anti-inflammatory properties. However, its effects on spermatogenic defects and testicular injury as well as the underlying mechanisms are still unclear. This study aims to investigate the protective effect of SRG on testis damage in zebrafish and explore its potential molecular events. Zebrafish testicular injury was induced by exposure to bisphenol A (BPA) (3000 µg/L) for two weeks. Fish were treated with intraperitoneal injection of SRG at different doses (5 and 50 mg/kg bodyweight) for two more weeks under BPA induction. Subsequently, the testis and sperm were collected for morphological, histological, biochemical and gene expression examination. It was found that the administration of SRG resulted in a significant protection from BPA-caused impact on sperm concentration, morphology, motility, fertility rate, testosterone level, spermatogenic dysfunction and resulted in increased apoptotic and reactive oxygen species' levels. Furthermore, testicular transcriptional profiling alterations revealed that the regulation of inflammatory response and oxidative stress were generally enriched in differentially expressed genes (DEGs) after SRG treatment. Additionally, it was identified that SRG prevented BPA-induced zebrafish testis injury through upregulation of fn1a, krt17, fabp10a, serpina1l and ctss2. These results indicate that SRG alleviated spermatogenic defects and testicular injury by suppressing oxidative stress and inflammation in male zebrafish.

Diverse functions of cytochrome c in cell death and disease.

The redox-active protein cytochrome c is a highly positively charged hemoglobin that regulates cell fate decisions of life and death. Under normal physiological conditions, cytochrome c is localized in the mitochondrial intermembrane space, and its distribution can extend to the cytosol, nucleus, and extracellular space under specific pathological or stress-induced conditions. In the mitochondria, cytochrome c acts as an electron carrier in the electron transport chain, facilitating adenosine triphosphate synthesis, regulating cardiolipin peroxidation, and influencing reactive oxygen species dynamics. Upon cellular stress, it can be released into the cytosol, where it interacts with apoptotic peptidase activator 1 (APAF1) to form the apoptosome, initiating caspase-dependent apoptotic cell death. Additionally, following exposure to pro-apoptotic compounds, cytochrome c contributes to the survival of drug-tolerant persister cells. When translocated to the nucleus, it can induce chromatin condensation and disrupt nucleosome assembly. Upon its release into the extracellular space, cytochrome c may act as an immune mediator during cell death processes, highlighting its multifaceted role in cellular biology. In this review, we explore the diverse structural and functional aspects of cytochrome c in physiological and pathological responses. We summarize how posttranslational modifications of cytochrome c (e.g., phosphorylation, acetylation, tyrosine nitration, and oxidation), binding proteins (e.g., HIGD1A, CHCHD2, ITPR1, and nucleophosmin), and mutations (e.g., G41S, Y48H, and A51V) affect its function. Furthermore, we provide an overview of the latest advanced technologies utilized for detecting cytochrome c, along with potential therapeutic approaches related to this protein. These strategies hold tremendous promise in personalized health care, presenting opportunities for targeted interventions in a wide range of conditions, including neurodegenerative disorders, cardiovascular diseases, and cancer.

Dysregulation of iron transport-related biomarkers in blood leukocytes is associated with poor prognosis of early trauma.

Objective: The early targeted and effective diagnosis and treatment of severe trauma are crucial for patients' outcomes. Blood leukocytes act as significant effectors during the initial inflammation and activation of innate immunity in trauma. This study aims to identify hub genes related to patients' prognosis in blood leukocytes at the early stages of trauma. Methods: The expression profiles of Gene Expression Omnibus (GEO) Series (GSE) 36809 and GSE11375 were downloaded from the GEO database. R software, GraphPad Prism 9.3.1 software, STRING database, and Cytoscape software were used to process the data and identify hub genes in blood leukocytes of early trauma. Results: Gene Ontology (GO) analysis showed that the differentially expressed genes (DEGs) of blood leukocytes at the early stages of trauma (0-4 h, 4-8 h, and 8-12 h) were mainly involved in neutrophil activation and neutrophil degranulation, neutrophil activation involved in immune response, neutrophil mediated immunity, lymphocyte differentiation, and cell killing. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the DEGs were mainly involved in Osteoclast differentiation and Hematopoietic cell lineage. Sixty-six down-regulated DEGs and 148 up-regulated DEGs were identified and 37 hub genes were confirmed by Molecular Complex Detection (MCODE) of Cytoscape. Among the hub genes, Lipocalin 2 (LCN2), Lactotransferrin (LTF), Olfactomedin 4 (OLFM4), Resistin (RETN), and Transcobalamin 1 (TCN1) were related to prognosis and connected with iron transport closely. LCN2 and LTF were involved in iron transport and had a moderate predictive value for the poor prognosis of trauma patients, and the AUC of LCN2 and LTF was 0.7777 and 0.7843, respectively. Conclusion: As iron transport-related hub genes in blood leukocytes, LCN2 and LTF can be used for prognostic prediction of early trauma.

Three cases of collared owlet depredation on the green-backed tit within nest boxes.

The main cause of the reproductive failure of cavity-nesting birds is nest predation, even though cavity nests protect from numerous predators. To study the breeding biology of the green-backed tit (Parus monticolus) and to promote the reproduction of some other avian cavity breeders, we placed 245 nest boxes in the Zixi Mountain, southwest China. We collected breeding data by regularly checking the artificial nest boxes, three cases of green-backed tits being predated by collared owlet (Glaucidium brodiei) were confirmed by the video recordings. Larger mammals, chipmunks, squirrels, sparrowhawks, jays, and snakes have been identified as common predators of cavity-nesting birds in high-latitude regions of the northern hemisphere. Limited research in tropical and subtropical regions, particularly in Asia, has demonstrated squirrels and snakes as common predators of cavity-nesting birds. A gap in avian predators to cavity-nesting birds exists in the current knowledge. Hence the three cases of collared owlet's depredation reveal a new danger to green-backed tits, broadening our knowledge of the dynamics of cavity-nesting birds. In all three cases, the artificial box's entrance hole was only 5 × 5 cm in size and has not been expanded or poked. These findings provided evidence that the collared owlet is the predator of nestlings and adult green-backed tit breeding in artificial boxes, which emphasized a reevaluation of predator-prey interactions. Therefore, for effective breeding of the green-backed tit, we suggest to choose a hole size of 3 × 3 cm that is appropriate for its body size.