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 investigate the recognition of the post competency index system among rural general practice assistant physicians and its influencing factors.Methods:This study was a cross-sectional survey. A questionnaire survey on the recognition of post competency index system was conducted from October 2020 to September 2021 among rural general practice assistant physicians from 10 provinces/municipalities selected by stratified cluster sampling method. The recognition of rural general practice assistant physicians at all levels of indexs and the factors influencing recognition were analyzed.Results:A total of 1 123 questionnaires were distributed and 1 024 valid ones were collected with a recovery rate of 91.18%. Of the 1 024 respondents, 529 were male(51.7%) and 435 were aged 40-49 years(42.5%), which was the highest proportion by age group. The average overall recognition score of the index system was 4.41, and the scores of the primary indexes were 4.32-4.45. Three primary indexes had the highest recognition scores: professional competence, basic health care services, and interpersonal communication and teamwork. The recognition scores on the second level index were 4.18-4.61, and the proportion of recognition scores greater than 4 was over 80%. There were significant differences in the recognition scores of the index system among assistant physicians with different working years, educational background, professional title and work unit ( F/H=6.41, 14.83, 12.45, 7.53, P<0.01). Educational background(associate degree: B=0.091, P=0.015; bachelor degree and above: B=0.196, P<0.001) and professional title(intermediate professional title and above: B=-0.234, P<0.001) were the independent factors influencing the recognition degree of the index system for rural general practice assistant physicians. Conclusions:The post competency index system is generally recognized by rural general practice assistant physician, and academic qualifications and professional title status may influence its recognition.

Objective:To evaluate the application feasibility of the post competency index system of rural general practice assistant physicians, and to analyze and compare the post competency of rural general practice assistant physicians with different characteristics through a survey among rural general practice assistant physicians in China.Methods:This study was a cross-sectional study. A questionnaire survey was conducted among rural general practice assistant physicians in 10 provinces/municipalities selected by stratified cluster sampling method from October 2020 to September 2021. The post competency scores were self-evaluated based on the post competency index system of rural general practice assistant physicians. The results of the survey were analyzed and the application feasibility of the index system was evaluated.Results:A total of 1 123 questionnaires were distributed and 1 024 valid questionnaires were returned with a recovery rate of 91.2%. Of the 1 024 respondents, 529 were males (51.7%), 435 were aged 40-49 years (42.5%), 434 had a secondary school education or less (42.4%), and 531 were junior practitioners (51.9%). The Cronbach′s α coefficient of the overall questionnaire was 0.987, and the Cronbach′s α coefficient of the first level index ranged from 0.897 to 0.974. The cumulative variance contribution rate of exploratory factor analysis was 72.012%. The confirmatory factor analysis showed χ2/ df=3.926, RMSEA=0.076, CFI=0.858, IFI=0.859, indicating that the model fit was basically good. The average self-evaluation scores of the first level index ranged from 3.95 to 4.25, and the average self-evaluation scores of the second level index ranged from 3.74 to 4.36. There were significant differences in self-evaluation scores of post competency among rural general practice assistant physicians with different working years, professional titles, working units and economic regions( F=4.67, 10.54, 22.16 and 20.90,all P<0.05). People with low self-evaluation scores of post competency had the following characteristics: working 10-19 years, intermediate or above titles, working in community health service centers, located in the eastern region.People with high self-evaluation scores of post competency had the following characteristics: primary professional title, working in the village clinic. Conclusion:The competency index system of rural general practice assistant physicians has good application feasibility,and it can be used to evaluate post competency for the education and training of rural general practice assistant physicians.

ObjectiveTo evaluate the effectiveness and safety of Qiaoqi Formula （翘芪组方） for mild influenza. MethodsA randomized controlled study was designed， recruiting 74 patients with mild influenza， who were randomly divided into trial group and control group. The trial group took Qiaoqi Formula orally， 40ml each time， twice a day； the control group took Lianhua Qingwen Capsules （连花清瘟胶囊） orally， 1.4 g each time， three times a day. Both groups were treated for 3 consecutive days and follow-up for 4 consecutive days after treatment. The time for fever reduction including onset of fever reduction， complete fever reduction time， fever reduction rates at 24， 48 and 72 hours， improvement of influenza symptoms， total traditional Chinese medicine （TCM） symptom score， and safety indicators in two groups after treatment were recorded. ResultsSixty-five patients were ultimately included， including 36 in the trial group and 29 in the control group. Onset time of fever reduction in the trial group was （15.49±23.47） h， the complete fever reduction time （21.37±30.06）h， and the 24 h， 48 h， 72 h， fever reduction rate was 77.14%， 88.57%， 91.42% respectively. The above indicators of the control group showed as （17.58±20.38）h， （24.30±21.87）h， 61.29%， 90.32%， 96.77% respectively， with no statistically significant differences （P＞0.05）. On the 7th day after treatment， the total score of TCM syndromes in trial group and control group decreased compared to those before treatment （P＜0.05）. There was no statistically significant difference in the cure rate， significant effective rate， effective rate， and total effective rate of TCM syndromes between groups （P＞0.05）. On the 4th day， the lymphocyte ratio of patients in the control group was higher than before treatment， while alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， urea， and creatinine of both groups before and after treatment were within the normal range. The main adverse reactions in both groups were mild headache and dizziness， and no serious adverse reactions observed. ConclusionThe therapeutic effect of Qiaoqi Formula in treating mild influenza is equivalent to Lianhua Qingwen Capsules， which can shorten the fever reduction time， improve clinical symptoms， and no adverse events observed during the study.

Purpose: The 21-gene recurrence score (RS) can guide adjuvant chemotherapy decisions in the multidisciplinary treatment (MDT) of patients with early breast cancer. This study aimed to evaluate the influence of the 21-gene RS assay on patient’ compliance with MDT and its association with disease outcomes. Methods: Patients diagnosed with pN0-1, hormone receptor-positive, human epidermal growth factor receptor-2-negative breast cancer between January 2013 and June 2019 were enrolled. A logistic regression model was used to identify parameters associated with treatment adherence. Prognostic indicators were evaluated using the Cox proportional hazard models. Results: After the assay, patients were less likely to violate the treatment plan (14.9% vs. 23.1%, p < 0.001), and higher compliance rates were observed for chemotherapy (p = 0.042), radiotherapy (p = 0.012), and endocrine therapy (p < 0.001). Multivariable analysis demonstrated that the 21-gene RS assay (odds ratio [OR], 1.43; 95% confidence interval [CI], 1.09–1.88; p = 0.009) was independently associated with MDT compliance. Moreover, compliance with MDT was independently associated with better disease-free survival (hazard ratio, 0.43; 95% CI, 0.29–0.64; p < 0.001), regardless of the 21-gene RS assay (interaction p = 0.842). Conclusion The 21-gene RS assay improved the MDT compliance rate in patients with early breast cancer. Adherence to MDT is associated with a better prognosis.

Nerve regeneration in adult mammalian spinal cord is poor because of the lack of intrinsic regeneration of neurons and extrinsic factors - the glial scar is triggered by injury and inhibits or promotes regeneration. Recent technological advances in spatial transcriptomics (ST) provide a unique opportunity to decipher most genes systematically throughout scar formation, which remains poorly understood. Here, we first constructed the tissue-wide gene expression patterns of mouse spinal cords over the course of scar formation using ST after spinal cord injury from 32 samples. Locally, we profiled gene expression gradients from the leading edge to the core of the scar areas to further understand the scar microenvironment, such as neurotransmitter disorders, activation of the pro-inflammatory response, neurotoxic saturated lipids, angiogenesis, obstructed axon extension, and extracellular structure re-organization. In addition, we described 21 cell transcriptional states during scar formation and delineated the origins, functional diversity, and possible trajectories of subpopulations of fibroblasts, glia, and immune cells. Specifically, we found some regulators in special cell types, such as Thbs1 and Col1a2 in macrophages, CD36 and Postn in fibroblasts, Plxnb2 and Nxpe3 in microglia, Clu in astrocytes, and CD74 in oligodendrocytes. Furthermore, salvianolic acid B, a blood-brain barrier permeation and CD36 inhibitor, was administered after surgery and found to remedy fibrosis. Subsequently, we described the extent of the scar boundary and profiled the bidirectional ligand-receptor interactions at the neighboring cluster boundary, contributing to maintain scar architecture during gliosis and fibrosis, and found that GPR37L1_PSAP, and GPR37_PSAP were the most significant gene-pairs among microglia, fibroblasts, and astrocytes. Last, we quantified the fraction of scar-resident cells and proposed four possible phases of scar formation: macrophage infiltration, proliferation and differentiation of scar-resident cells, scar emergence, and scar stationary. Together, these profiles delineated the spatial heterogeneity of the scar, confirmed the previous concepts about scar architecture, provided some new clues for scar formation, and served as a valuable resource for the treatment of central nervous system injury.
Mice ; Animals ; Gliosis/pathology* ; Cicatrix/pathology* ; Spinal Cord Injuries ; Astrocytes/metabolism* ; Spinal Cord/pathology* ; Fibrosis ; Mammals ; Receptors, G-Protein-Coupled

Biapenem is a carbapenem antibiotic， and can be used for the treatment of sepsis， pneumonia， lung abscess， chronic respiratory lesions secondary infection， complex urinary tract infection and pyelonephritis， etc. This article reviewed the studies on the pharmacokinetics， pharmacodynamics and therapeutic drug monitoring （TDM） of biapenem. The pharmacokinetic parameters of biapenem are not significantly different in healthy subjects， and there is no accumulation after multiple doses of biapenem. However， there are large differences in pharmacokinetic parameters in patients with severe disease and patients with abnormal renal function compared with healthy subjects， which leads to conventional treatment regimens not achieving the desired outcome. In terms of pharmacodynamics， biapenem can improve the rate of reaching the target value by increasing the frequency of administration and prolonging the infusion time. For patients with anuria in end-stage renal disease， dosing intervals can be extended to avoid drug accumulation. However, for patients with severe infection, a daily dose of 1.2 g still can not control infections caused by Acinetobacter baumannii or Pseudomonas aeruginosa, which limits its use in patients with severe disease. It is recommended to implement TDM in severe patients and patients with abnormal renal function， and explore the best dosing regimen for biapenem in combination with pharmacokinetic models to ensure that the time that the free blood concentration of biapenem remains above minimum inhibitory concentration as a percentage of the time between doses （%fT＞MIC） is within the effective range，so that biapenem can exert a greater efficacy in severe patients and patients with abnormal renal function. For medical institutions that cannot carry out TDM， the efficacy of biapenem can be maximized by increasing the frequency of administration and prolonging the infusion time. For infections caused by P. aeruginosa， A. baumannii and Serratia marcescens with high drug resistance rates， it is recommended to combine or replace other antibiotics.

During coronavirus disease 2019 epidemic, the treatment of severe trauma has been impacted. The Consensus on emergency surgery and infection prevention and control for severe trauma patients with 2019 novel corona virus pneumonia was published online on February 12, 2020, providing a strong guidance for the emergency treatment of severe trauma and the self-protection of medical staffs in the early stage of the epidemic. With the Joint Prevention and Control Mechanism of the State Council renaming "novel coronavirus pneumonia" to "novel coronavirus infection" and the infection being managed with measures against class B infectious diseases since January 8, 2023, the consensus published in 2020 is no longer applicable to the emergency treatment of severe trauma in the new stage of epidemic prevention and control. In this context, led by the Chinese Traumatology Association, Chinese Trauma Surgeon Association, Trauma Medicine Branch of Chinese International Exchange and Promotive Association for Medical and Health Care, and Editorial Board of Chinese Journal of Traumatology, the Chinese expert consensus on emergency surgery for severe trauma and infection prevention during coronavirus disease 2019 epidemic ( version 2023) is formulated to ensure the effectiveness and safety in the treatment of severe trauma in the new stage. Based on the policy of the Joint Prevention and Control Mechanism of the State Council and by using evidence-based medical evidence as well as Delphi expert consultation and voting, 16 recommendations are put forward from the four aspects of the related definitions, infection prevention, preoperative assessment and preparation, emergency operation and postoperative management, hoping to provide a reference for severe trauma care in the new stage of the epidemic prevention and control.

BACKGROUND: Repair of long-distance peripheral nerve defects remains an important clinical problem. Nerve grafts incorporated with extracellular vesicles (EVs) from various cell types have been developed to bridge peripheral nerve defects. In our previous research, EVs obtained from skin-derived precursor Schwann cells (SKP-SC-EVs) were demonstrated to promote neurite outgrowth in cultured cells and facilitate nerve regeneration in animal studies. METHODS: To further assess the functions of SKP-SC-EVs in nerve repair, we incorporated SKP-SC-EVs and Matrigel into chitosan nerve conduits (EV-NG) for repairing a 15-mm long-distance sciatic nerve defect in a rat model. Behavioral analysis, electrophysiological recording, histological investigation, molecular analysis, and morphometric assessment were carried out. RESULTS: The results revealed EV-NG significantly improved motor and sensory function recovery compared with nerve conduits (NG) without EVs incorporation. The outgrowth and myelination of regenerated axons were improved, while the atrophy of target muscles induced by denervation was alleviated after EVs addition. CONCLUSION Our data indicated SKP-SC-EVs incorporation into nerve grafts represents a promising method for extended peripheral nerve damage repair.

Objective:To survey the competency characteristics of rural general practitioners using behavioral event interview.Methods:The competency characteristics of rural general practitioners were extracted by literature research method, and the behavior event interviews were conducted with 18 rural general practitioners nationwide from January to April 2019. According to the interview results, the items were modified and refined. On this basis, the benchmark competencies and excellent competencies of rural general practitioners were extracted.Results:A total of 45 885-second audio recordings and 174 523-word transcripts were obtained from the interviews. Eight identification competencies and 10 benchmark competencies were extracted. Identification competencies included "professionalism, altruism and dedication, love and compassion, visiting a patient at home, establishing good relationships, achievement-oriented, having expertise, and mastering community population dynamics and health status actively". Benchmark competencies included "diagnosis and treatment of common diseases, basic medical skills, learning consciousness and ability, solving medical problems with traditional Chinese medicine, health care for focal groups, rational administration of drug, referral services, managing chronic non communicable diseases, understanding health care system and related policies, and emergency treatment ability".Conclusions:This study preliminarily obtains the identification competencies and benchmark competencies of rural general practitioners. Managers can improve the service quality of rural general practitioners according to these elements.