Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

Objective:To analyze the acute poison epidemic and provide evidence for developing prevention and control strategies for acute poisoning.Methods:A retrospective analysis was conducted on acute poisoning cases collected from 2016 to 2022 in a health emergency information platform for acute poisoning accidents. The cases were grouped according to the distribution of poisoning occurrence time, geographic distribution, demographic distribution, types of toxicants, poisoning causes, and outcomes. Data were organized and analyzed using Excel 2016 and R 4.2.3.Results:A total of 95 754 acute poisoning cases were included in this study. The primary toxicants were pesticides, drugs, and industrial/household chemicals, accounting for 30.4%, 22.4%, and 20.4% of the total cases, respectively. Acute poisoning occurred throughout the year, with the highest frequency from June to August, accounting for 31.9%. The seasonal distribution varied among different types of toxicants. Except for plant poisoning, which showed a bimodal distribution, the other poisonings showed an unimodal distribution. There was a strong seasonality in fungal poisoning, which peaked in July. There was an obvious seasonality in animal poisoning, with a peak in August. The proportion of biological poisonings in the southwest region was higher than in other regions, including plants, animals, and fungi. There were more females than males, and their education level was mainly junior high school and below (35.2%). The main occupation was farmers (34.2%), and the main causes of poisoning were accidents and suicides. The case fatality rate of all poisoning cases was 1.24%. Pesticide poisoning was the most common type, and chlorfenapyr (11.68%), Diquat (7.23%), and paraquat (7.05%) ranked as the top three toxicants.Conclusions:The occurrence of acute poisoning has an obvious seasonal trend, and the toxicant spectrum of different regions and populations is different. A comprehensive poisoning surveillance system can provide a better understanding of the occurrence of poisonings, and facilitate the formulation of more scientifically precise poisoning prevention and control strategies.

ObjectiveTo explore the therapeutic effect and mechanism of Guipitang on rats with myocardial ischemia. MethodFifty SD rats were divided into five groups： a control group， a model group， low and high-dose Guipitang （7.52， 15.04 g·kg^-1） groups， and a trimetazidine group （0.002 g·kg^-1）. By intragastric administration of vitamin D₃ and feeding rats with high-fat forage and injecting isoproterenol， the rat model of myocardial ischemia was established. After drug treatment of 15 d， an electrocardiogram （ECG） was performed to analyze the degree of myocardial injury. A fully automatic biochemical analyzer was used to detect the changes in the serum levels of total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C）. Hematoxylin-eosin （HE） staining and Masson staining were used to observe myocardial histopathological changes. TdT-mediated dUTP nick end labeling （TUNEL） staining was used to detect cardiomyocyte apoptosis. Western blot was adopted to detect the protein levels of extracellular signal-regulated kinase 1/2 （ERK1/2）， phospho-ERK1/2 （p-ERK1/2）， p38 mitogen-activated protein kinase （p38 MAPK）， phospho-p38 MAPK （p-p38 MAPK）， B-cell lymphoma-2 （Bcl-2）-associated X （Bax）， Bcl-2， and cleaved cysteine aspartate proteolytic enzyme （cleaved Caspase-3）. ResultCompared with the control group， the ECG S-T segment decreased in the model group. The serum levels of TC， TG， and LDL-C were increased significantly （P<0.05）. The arrangement of myocardial tissue was disordered， and the proportion of cardiomyocyte apoptosis increased. The protein levels of cleaved Caspase-3， Bax， and p-p38 MAPK in the heart were increased， and the Bcl-2 expression was decreased （P<0.05）. Compared with the model group， the S-T segment downward shift was restored in the low and high-dose Guipitang groups and trimetazidine group， and the levels of TC， TG， and LDL-C were decreased. The protein expression of cleaved Caspase-3 and Bax in the heart dropped， and p-p38 MAPK and p-ERK1/2 protein expressions increased significantly （P<0.05）. The degree of myocardial injury was alleviated， and the proportion of cardiomyocyte apoptosis decreased. Bcl-2 protein expression was increased significantly in the low-dose Guipitang group （P<0.05）. ERK1/2 and p38 MAPK proteins had no significant difference among different groups. ConclusionGuipitang could alleviate myocardial injury and inhibit cardiomyocyte apoptosis in rats by activating the expression of ERK1/2 and p38 MAPK.

Objective To explore the predictive value of 3D quantitative shape features based on enhanced CT for disease-free survival(DFS)of primary gastric gastrointestinal stromal tumors(GIST)patients receiving curative resection.Methods From January 2010 to December 2016,a total of 154 patients with primary gastric GIST who received curative resection in Zhongshan Hospital,Fudan University were retrospectively collected.The conventional CT imaging features and 3D quantitative shape features of tumors were evaluated,and the influencing factors of DFS were analyzed by univariate and multivariate Cox regression.Cut-off values were extracted,and Kaplan-Meier curves were used for survival analysis.Results Maximum 3D diameter(HR=1.829,95% CI 1.389-2.408,P＜0.001)and spherical disproportion(HR=2.153,95% CI 1.474-3.146,P＜0.001)were independent risk factors for DFS in primary gastric GIST.Kaplan-Meier curves showed that patients with maximum 3D diameter＞90.5 mm and spherical disproportion＞1.5 showed shorter DFS after curative resection(P＜0.001).Conclusion Maximum 3D diameter and spherical disproportion based on preoperative enhanced CT are valuable in the diagnosis of postoperative DFS in primary gastric GIST.

Objective:To investigate the effects of cerium oxide nanoenzyme-gelatin methacrylate anhydride (GelMA) hydrogel (hereinafter referred to as composite hydrogel) in the repair of infected full-thickness skin defect wounds in mice.Methods:This study was an experimental study. Cerium oxide nanoenzyme with a particle size of (116±9) nm was prepared by hydrothermal method, and GelMA hydrogel with porous network structure and good gelling performance was also prepared. The 25 μg/mL cerium oxide nanoenzyme which could significantly promote the proliferation of human skin fibroblasts and had high superoxide dismutase activity was screened out. It was added to GelMA hydrogel to prepare composite hydrogel. The percentage of cerium oxide nanoenzyme released from the composite hydrogel was calculated after immersing it in phosphate buffer solution (PBS) for 3 and 7 d. The red blood cell suspension of mice was divided into PBS group, Triton X-100 group, cerium oxide nanoenzyme group, GelMA hydrogel group, and composite hydrogel group, which were treated with corresponding solution. The hemolysis of red blood cells was detected by microplate reader after 1 h of treatment. The bacterial concentrations of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli were determined after being cultured with PBS, cerium oxide nanoenzyme, GelMA hydrogel, and composite hydrogel for 2 h. The sample size in all above experiments was 3. Twenty-four 8-week-old male BALB/c mice were taken, and a full-thickness skin defect wound was prepared in the symmetrical position on the back and infected with MRSA. The mice were divided into control group without any drug intervention, and cerium oxide nanoenzyme group, GelMA hydrogel group, and composite hydrogel group applied with corresponding solution, with 6 mice in each group. The wound healing was observed on 3, 7, and 14 d after injury, and the remaining wound areas on 3 and 7 d after injury were measured (the sample size was 5). The concentration of MRSA in the wound exudation of mice on 3 d after injury was measured (the sample size was 3), and the blood flow perfusion in the wound of mice on 5 d after injury was observed using a laser speckle flow imaging system (the sample size was 6). On 14 d after injury, the wound tissue of mice was collected for hematoxylin-eosin staining to observe the newly formed epithelium and for Masson staining to observe the collagen situation (the sample size was both 3). Results:After immersion for 3 and 7 d, the release percentages of cerium oxide nanoenzyme in the composite hydrogel were about 39% and 75%, respectively. After 1 h of treatment, compared with that in Triton X-100 group, the hemolysis of red blood cells in PBS group, GelMA hydrogel group, cerium oxide nanoenzyme group, and composite hydrogel group was significantly decreased ( P<0.05). Compared with that cultured with PBS, the concentrations of MRSA and Escherichia coli cultured with cerium oxide nanoenzyme, GelMA hydrogel, and composite hydrogel for 2 h were significantly decreased ( P<0.05). The wounds of mice in the four groups were gradually healed from 3 to 14 d after injury, and the wounds of mice in composite hydrogel group were all healed on 14 d after injury. On 3 and 7 d after injury, the remaining wound areas of mice in composite hydrogel group were (29±3) and (13±5) mm 2, respectively, which were significantly smaller than (56±12) and (46±10) mm 2 in control group and (51±7) and (38±8) mm 2 in cerium oxide nanoenzyme group (with P values all <0.05), but was similar to (41±5) and (24±9) mm 2 in GelMA hydrogel group (with P values both >0.05). On 3 d after injury, the concentration of MRSA on the wound of mice in composite hydrogel group was significantly lower than that in control group, cerium oxide nanoenzyme group, and GelMA hydrogel group, respectively (with P values all <0.05). On 5 d after injury, the volume of blood perfusion in the wound of mice in composite hydrogel group was significantly higher than that in control group, cerium oxide nanoenzyme group, and GelMA hydrogel group, respectively ( P<0.05). On 14 d after injury, the wound of mice in composite hydrogel group basically completed epithelization, and the epithelization was significantly better than that in the other three groups. Compared with that in the other three groups, the content of collagen in the wound of mice in composite hydrogel group was significantly increased, and the arrangement was also more orderly. Conclusions:The composite hydrogel has good biocompatibility and antibacterial effect in vivo and in vitro. It can continuously sustained release cerium oxide nanoenzyme, improve wound blood perfusion in the early stage, and promote wound re-epithelialization and collagen synthesis, therefore promoting the healing of infected full-thickness skin defect wounds in mice.

Objective:To explore the moderating mechanism of mindfulness on perceived stress and job burnout of medical staff.Methods:From November 2020 to March 2022, 1626 medical staff were investigated by questionnaires. Chinese Version Perceived Stress Scale (CPSS), Maslach Burnout Inventory-Human Service Survey (MBI-HSS) and Mindful Attention Awareness Scale (MAAS) were used to evaluate the perceived stress, three dimensions of job burnout[depersonalization (DP), emotional exhaustion (EE), personal accomplishment (PA) ], and mindfulness level of medical staff. The Spearman rank correlation analysis was used to analyze the correlation between mindfulness and perceived stress, job burnout of medical staff. And the SPSS PROCESS macro program was used to test the moderating effect of mindfulness on perceived stress and job burnout.Results:Among the 1626 medical staff, 57.38% had perceived stress with health risk (933/1626), and 63.84% (1038/1626) had job burnout, among them, with 618 (38.01%), 274 (16.85%), and 146 (8.98%) experiencing mild, moderate and severe job burnout, respectively. The scores of mindfulness among medical staff were significantly correlated with perceived stress and various dimensions of job burnout (EE, DP and PA) ( rs=-0.155, -0.351, -0.315, 0.307, P<0.001). Mindfulness had a moderating effect between perceived stress and job burnout, which was achieved through three moderating pathways: perceived stress-mindfulness-EE, perceived stress-mindfulness-DP and perceived stress-mindfulness-PA (Δ R2=0.073, 0.06, 0.006, P<0.001) . Conclusion:Medical staff have a high level of job burnout, mindfulness plays a moderating role between perceived stress and job burnout. Specific measures can be taken to improve the level of mindfulness in medical staff, so as to prevent and alleviate job burnout of medical staff.

{L-End}Objective To explore the chain mediating effect of team psychological safety atmosphere and professional calling in the relationship between humble leadership and voice behavior. {L-End}Methods A total of 812 nurses from five medical institutions in Guangdong Province were selected as the research subjects using the convenience sampling method. The Team Psychological Safety Atmosphere Scale, the Professional Calling Scale, the Hospital Version of Humble Leadership Scale and the Voice Behavior Scale were used to investigate the perceived team psychological safety atmosphere and sense of professional calling, the perceived humble leadership style and voice behavior of the nurses. {L-End}Results The score of team psychological safety atmosphere, professional calling, humble leadership, and voice behavior of 812 nurses was (58.0±7.6), (44.6±8.0), (50.4±9.2) and (37.0±5.9), respectively, with the scoring rate of 72.5%, 74.3%, 80.0%, and 74.0%, respectively. Bootstrap analysis showed that humble leadership could positively influence voice behavior ［standardized effect value (β)=0.28, 95% confidence interval (CI) 0.20-0.35］. Humble leadership could influence voice behavior through the separate mediating effects of team psychological safety atmosphere (β=0.13, 95%CI 0.09-0.18), professional calling (β=0.07, 95%CI 0.04-0.11), and the chain mediating effect of team psychological safety atmosphere and professional calling(β=0.06, 95%CI 0.04-0.08). {L-End}Conclusion The voice behavior of clinical nurses is at a moderately high level. Humble leadership can directly influence voice behavior and can influence voice behavior through the mediating effects of team psychological safety atmosphere and professional calling. Nursing managers can inspire the work passion of clinical nurses by changing their own leadership style and creating a good atmosphere for voicing opinions.

Objective:To explore the moderating mechanism of mindfulness on perceived stress and job burnout of medical staff.Methods:From November 2020 to March 2022, 1626 medical staff were investigated by questionnaires. Chinese Version Perceived Stress Scale (CPSS), Maslach Burnout Inventory-Human Service Survey (MBI-HSS) and Mindful Attention Awareness Scale (MAAS) were used to evaluate the perceived stress, three dimensions of job burnout[depersonalization (DP), emotional exhaustion (EE), personal accomplishment (PA) ], and mindfulness level of medical staff. The Spearman rank correlation analysis was used to analyze the correlation between mindfulness and perceived stress, job burnout of medical staff. And the SPSS PROCESS macro program was used to test the moderating effect of mindfulness on perceived stress and job burnout.Results:Among the 1626 medical staff, 57.38% had perceived stress with health risk (933/1626), and 63.84% (1038/1626) had job burnout, among them, with 618 (38.01%), 274 (16.85%), and 146 (8.98%) experiencing mild, moderate and severe job burnout, respectively. The scores of mindfulness among medical staff were significantly correlated with perceived stress and various dimensions of job burnout (EE, DP and PA) ( rs=-0.155, -0.351, -0.315, 0.307, P<0.001). Mindfulness had a moderating effect between perceived stress and job burnout, which was achieved through three moderating pathways: perceived stress-mindfulness-EE, perceived stress-mindfulness-DP and perceived stress-mindfulness-PA (Δ R2=0.073, 0.06, 0.006, P<0.001) . Conclusion:Medical staff have a high level of job burnout, mindfulness plays a moderating role between perceived stress and job burnout. Specific measures can be taken to improve the level of mindfulness in medical staff, so as to prevent and alleviate job burnout of medical staff.

Objective : To explore the epicardial adipose tissue ( EAT) of patients with coronary heart disease , KLF7 stimulates macrophages to secrete inflammatory factors and promotes the differentiation and maturation of adipocytes through the NF⁃κB signaling pathway , and to clarify the mechanism of KLF7 in the occurrence and development of CAD. Methods : 30 patients with coronary heart disease ( CAD group) and 30 patients without coronary heart disease (non⁃CAD group) were collected , and general data and biochemical indicators were collected. qRT⁃PCR was used to detect the expression levels of KLF7 , APN , IL⁃6 , and TNF⁃α mRNA in EAT. Human THP⁃1 cells were cultured in vitro and induced into M1 type macrophages and 3T3 ⁃L1 preadipocytes. The cells were divied into 3 groups : KLF7 up⁃regulated group ( transfected with KLF7 mimic) , KLF7 down⁃regulated group ( transfected with siRNA knockdown KLF7 ) , NC group ( transfected oligopeptide sequence) , transfected two kinds of cells. qRT⁃PCR was used to detect the expression of APN , MCP⁃1 , IL⁃6 and TNF⁃α mRNA in M1 type macrophages , and the protein expression levels of key factors in the NF⁃κB signaling pathway were detected by Westren blot. The qRT⁃PCR method was used to detect APN , KLF4 , IL⁃6 , MCP⁃1 mRNA and adipocyte differentiation marker peroxisome proliferator⁃activated receptor⁃γ (PPARγ) in 3T3 ⁃L1 preadipocytes 24 h after transfection. CCAAT/enhancer binding protein α (C/EBPα ) , fatty acid binding protein 4 (FABP4) mRNA relative expression levels , and Westren blot was used to detect protein expression levels. Results : Compared with the non⁃CAD group , the expression of CAD group decreased , APN decreased , IL⁃6 and TNF⁃α increased significantly , and the difference was statistically significant (P < 0. 01) . KLF7 was highly expressed in human THP⁃1 derived M1 macrophages induced by inflammatory stimuli (LPS) . In M1 macrophages derived from human THP⁃1 , knocking down KLF7 could inhibit the release of inflammatory factors. Transfection with KLF7 ⁃siRNA could significantly inhibit LPS⁃induced phosphorylation of JNK⁃MAPKs , the level of p⁃p65 and the activation of p ⁃IκBa (P < 0. 05) . In 3T3 ⁃L1 preadipocytes , upregulation of KLF7 increased the expression of adipocyte differentiation markers PPARγ , C/EBPa , FABP4 mRNA , and promoted the differentiation of 3T3 ⁃L1 preadipocytes into adipocytes (P < 0. 05) . Conclusion The expression of KLF7 in EAT in CAD patients increases. KLF7 activates the activation of macrophages mediated by the JNK⁃NF⁃KB signaling pathway in EAT , promotes inflammation in EAT in CAD patients , and promotes the differentiation and maturation of adipocytes , thereby promoting the development of CAD. It indicates that KLF7 may be a potential therapeutic target for cardiovascular diseases (such as CAD) .

Objective:To observe the effects of static 70° head-up tilted standing and of repeated body repositioning on hemodynamics in healthy young and middle-aged persons.Methods:The hemodynamics of 24 middle-aged and 23 younger persons were studied. Both groups were requested to perform static 70° head-up tilted standing and to repeatedly change their body position from 0° to 70° of tilt at a velocity of 1°/second for ten minutes in a random order. Before, between and after each test the subjects rested supine for ten minutes. Hemodynamic variables and blood pressure were recorded non-invasively.Results:The average heart rate (HR) increased significantly in both groups when rising from supine to the testing positions. In 70° tilted standing the average HR of the youth group, 84.0±9.5bpm, was significantly higher than that in the other position and that of the middle-aged group in the same position. The average HR of the middle-aged group in 70° tilted standing was also significantly higher than in the other position. Among the middle-aged group, the average stroke volume (SV) in the testing positions was significantly lower than when resting. Significant differences were observed in the average diastolic blood pressure (DBP) between the testing and rest positions for both groups, with the average DBP of the middle-aged group significantly higher than that of the youth group in all three positions. Among the youth group, the average SV, CO and systolic blood pressure (SBP) of the males were significantly higher than among the females in all of the different body positions.Conclusions:Young persons mainly rely on an increased heart rate to maintain cardiac output while middle-aged participants appear to achieve this through increased peripheral resistance. Repeated position changes have less impact on hemodynamics than 70° inclined standing, making it a safer and more stable training method. However, the long-term effects of such intervention need to be confirmed in further studies.