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*

N⁶-methyladenosine (m⁶A) modification plays a critical role in cell cycle regulation, while the mechanism of m⁶A in regulating mitosis remains underexplored. Here, we found that the total m⁶A modification level in cells increased during mitosis by the liquid chromatography-mass spectrometry/mass spectrometry and m⁶A dot blot assays. Silencing methyltransferase-like 3 (METTL3) or METTL14 results in delayed mitosis, abnormal spindle assembly, and chromosome segregation defects by the immunofluorescence. By analyzing transcriptome-wide m⁶A targets in HeLa cells, we identified polo-like kinase 1 (PLK1) as a key gene modified by m⁶A in regulating mitosis. Specifically, through immunoblotting and RNA pulldown, m⁶A modification inhibits PLK1 translation via YTH N⁶-methyladenosine RNA binding protein 1, thus mediating cell cycle homeostasis. Demethylation of PLK1 mRNA leads to significant mitotic abnormalities. These findings highlight the critical role of m⁶A in regulating mitosis and the potential of m⁶A as a therapeutic target in proliferative diseases such as cancer.
Humans ; Polo-Like Kinase 1 ; Cell Cycle Proteins/metabolism* ; Proto-Oncogene Proteins/metabolism* ; Protein Serine-Threonine Kinases/metabolism* ; Mitosis/physiology* ; HeLa Cells ; Adenosine/genetics* ; Methyltransferases/metabolism* ; RNA, Messenger/metabolism* ; RNA-Binding Proteins/metabolism*

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease affecting both upper and lower motor neurons (MNs) with large unmet medical needs. Multiple pathological mechanisms are considered to contribute to the progression of ALS, including neuronal oxidative stress and mitochondrial dysfunction. Honokiol (HNK) has been reported to exert therapeutic effects in several neurologic disease models including ischemia stroke, Alzheimer's disease and Parkinson's disease. Here we found that honokiol also exhibited protective effects in ALS disease models both in vitro and in vivo. Honokiol improved the viability of NSC-34 motor neuron-like cells that expressed the mutant G93A SOD1 proteins (SOD1-G93A cells for short). Mechanistical studies revealed that honokiol alleviated cellular oxidative stress by enhancing glutathione (GSH) synthesis and activating the nuclear factor erythroid 2-related factor 2 (NRF2)-antioxidant response element (ARE) pathway. Also, honokiol improved both mitochondrial function and morphology via fine-tuning mitochondrial dynamics in SOD1-G93A cells. Importantly, honokiol extended the lifespan of the SOD1-G93A transgenic mice and improved the motor function. The improvement of antioxidant capacity and mitochondrial function was further confirmed in the spinal cord and gastrocnemius muscle in mice. Overall, honokiol showed promising preclinical potential as a multiple target drug for ALS treatment.

The medical massive open online course (MOOC) quality certification standard is the reference for the implementation of medical MOOC quality monitoring. This thesis utilizes data mining, quantitative analysis, qualitative analysis and other methods, taking the comment text of the Chinese medicine courses on the Ai Course (MOOC of China Universities) platform as the data source out of which to extract certain potential indicators for the medical MOOC quality certification standard. Based on the analysis of the existing research results on MOOC quality evaluation domestic and abroad as well as the relevant policies of the Ministry of Education, a medical MOOC quality accreditation standard including 6 first-level indicators are formed, including course content, teaching design and so on. Expert scores and AHP method were used to calculate the weight coefficient of the quality certification standard. Finally, this paper explains the procedure of medical MOOC quality certification in college. This certification system can be used to conduct self-assessment and quality monitoring of medical MOOC in colleges and universities.

This study summarizes related studies on readiness for online teaching in China and globally and constructs an online teaching readiness scale for higher education teachers with reference to related studies on online teaching competency, blended teaching readiness, and influencing factors for online teaching. This scale includes five dimensions, i.e., belief, teaching readiness, technical readiness, online communication readiness, and institutional support. A questionnaire survey and statistical analyses were performed to investigate the rationality of the scale, and then the scale was modified. The results show that the indicators of the scale have a good degree of fitting, and this study provides a necessary standard for examining the online teaching readiness of higher education teachers and new ideas for online teaching in colleges and universities.

[Abstract] Objective: To investigate the effect of long non-coding RNA (lncRNA)-CCAT2 on the proliferation and cell cycle of cervical cancer cells. Methods: The expression of CCAT2 in 3 cervical cancer cell lines (HeLa, C-33A, and CaSki) was detected by qPCR and the cell line with the highest expression level was selected for subsequent experiments. CCAT2 overexpression and interference vectors were designed and synthesized. After transfection, qPCR was performed to detect the transfection efficiency. The cells were divided into 5 groups: control, sh-EV (empty vector), overExp-EV , sh-CCAT2, and overExp-CCAT2. MTT assay was performed to evaluate cell viability. Flow cytometry was performed to measure cell cycle. WB was performed to detect the expressions of Ki67, cyclin D1, and cyclin dependent kinase 4 (CDK4). Results: Among HeLa, C-33A, and CaSki cells, the highest expression of CCAT2 was found in CaSki cells. CCAT2 overexpression and interference vectors were successfully transfected into the CaSki cells. Compared with the control group, the cells viability and proliferation in the sh-CCAT2 group was significantly decreased (all P<0.01), the proportion of cells in the G1 phase was significantly increased (P<0.01), and the expression levels of Ki67, cyclin D1, and CDK4 were significantly decreased (all P<0.01). However, in the overExp-CCAT2 group, the cell proliferation was enhanced and the expression levels of Ki67, cyclin D1, and CDK4 were significantly increased (all P<0.01). Conclusion: CCAT2 affects proliferation and cell cycle of cervical cancer cells by regulating the expressions of their associated proteins.

Objective: To study the correlation between job satisfaction and work effectiveness of ICU specialist nurses, to clarify the baseline level and influence factors, to increase the efficiency and satisfaction of the work, and to stabilize the ICU nurse team, to improve the quality of care. Methods: The convenience sampling method was used to select 116 specialist nurses from the 3 three grade ICU hospitals in Guangzhou. Through the questionnaire survey of nurses′ general situation, work efficiency and job satisfaction, the methods of descriptive statistics, average number, standard deviation analysis, simple linear regression model analysis and multiple linear regression analysis were adopted to reveal the relationship between work effectiveness and job satisfaction of specialist nurses. Results: The overall empowerment of ICU specialist nurses was 3.37 ±0.68. In the 6 dimensions of work efficiency, the average score of formal authorization was the lowest (3.01±0.93) and that of informal authorization was the highest (3.73±0.77). Multiple linear regression analysis showed that job satisfaction of nurses increased linearly with the increase of the scores of formal authorization and informal authorization (t=2.32, 3.78,P < 0.05). The total score of satisfaction was 55.03± 8.30. The work status dimension score was 35.16± 5.94, in the middle level limit; the work interpersonal relationship dimension score was 19.87 ± 2.82, in the high level. The work status satisfaction lagged behind the work interpersonal relationship satisfaction. Simple linear regression model analysis showed that the total score of satisfaction increased linearly with the average score of work efficiency, showing a positive linear correlation (t=9.891,P < 0.01). The regression equation was: y=27.12+8.273. Conclusions Because of the linear positive correlation between job efficacy and total score of satisfaction, nursing administrators should fully grasp the various factors affecting job efficacy in clinical management, and take targeted measures to improve the work efficiency of ICU specialist nurses, and ultimately improve the job satisfaction of nurses.

In this study, eight-month-old ectomycorrhizae of Tuber borchii with Corylus avellana were synthesized to explore the influence of T. borchii colonization on the soil properties and the microbial communities associated with C. avellana during the early symbiotic stage. The results showed that the bacterial richness and diversity in the ectomycorrhizae were significantly higher than those in the control roots, whereas the fungal diversity was not changed in response to T. borchii colonization. Tuber was the dominant taxon (82.97%) in ectomycorrhizae. Some pathogenic fungi, including Ilyonectria and Podospora, and other competitive mycorrhizal fungi, such as Hymenochaete, had significantly lower abundance in the T. borchii inoculation treatment. It was found that the ectomycorrhizae of C. avellana contained some more abundant bacterial genera (e.g., Rhizobium, Pedomicrobium, Ilumatobacter, Streptomyces, and Geobacillus) and fungal genera (e.g., Trechispora and Humicola) than the control roots. The properties of rhizosphere soils were also changed by T. borchii colonization, like available nitrogen, available phosphorus and exchangeable magnesium, which indicated a feedback effect of mycorrhizal synthesis on soil properties. Overall, this work highlighted the interactions between the symbionts and the microbes present in the host, which shed light on our understanding of the ecological functions of T. borchii and facilitate its commercial cultivation.
Colon ; Corylus ; Fungi ; Magnesium ; Mycorrhizae ; Nitrogen ; Phosphorus ; Podospora ; Rhizobium ; Rhizosphere ; Soil ; Streptomyces

Objective: To investigate the application of clohexidine wipes in patients after liver transplantation and the effect of infection prevention. Methods: A total of 279 patients who received orthotopic liver transplantation in the first affiliated hospital of sun yat-sen university from January 2017 to December 2018 and were transferred to the intensive care department of our hospital after the surgery were selected as subjects. Among them, 145 patients who received liver transplantation between January and December 2017 were selected as the control group. From January to December, 2018, 134 patients after liver transplantation were enrolled as the intervention group, and were treated with clohexidine 2% gluconate wet wipes, and the hospital infection rate, infection rate, multiple drug-resistant bacteria infection rate, bath time consumption, and adverse reaction rate were compared between the two groups. Results: The hospital infection rate of the control group was 20.00% (29/145), the infection rate of patients was 37.93%(55/145), the infection rate of multiple drug-resistant bacteria was 25.52%(37/145), and the infection rate of the intervention group was 11.19% (15/134), 24.63% (33/134), 14.93% (20/134). The differences were statistically significant (χ²=4.065, 5.709, 4.806, P<0.05). The bath time of the control group was (16.70±1.42) minutes, and the intervention group was (14.07±1.53) minutes. Compared with the control group, the bath time of the intervention group was saved, and the difference was statistically significant (t=9.637, P<0.01).No discomfort symptoms such as disinfectant allergy were found in both groups. Conclusion The application of clohexidine 2% gluconate wet wipes in the wiping bath of patients after liver transplantation had a good effect, significantly reducing the hospital infection rate, saving the time of wiping bath.