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*

Recent studies have identified a missense mutation in the β1-receptor (ADRB1-A187V) that exerts a pronounced impact on human sleep, with a noted decrease in protein abundance in vivo. The administration of β-blockers is frequently associated with sleep disturbances in clinical settings. In this study, we assessed the influence of various β-blockers on sleep within mouse models. Our findings indicated that β-blockers could induce varying degrees of arousal, sleep disruption, and a decrease in REMS (rapid eye movement sleep). We examined the dose-dependent effects of metoprolol and nebivolol on both sleep and cardiac functionality in both wild-type and Adrb1-A187V mutant mice. Our data suggested that, in contrast to cardiac effects, higher doses of metoprolol are required to have noted impact on sleep. No genotype effect was observed with metoprolol in terms of sleep or cardiac function. In contrast, the mutant mice demonstrated increased sensitivity to nebivolol, which exacerbated sleep fragmentation and impeded the onset of REMS. This study is expected to provide some reference for minimizing the occurrence of sleep disorders and reducing the adverse reactions of drugs to the greatest extent.

OBJECTIVE To study the constituents from the dried stems and leaves of Illicium dunnianum Tutcher.METHODS The compounds were isolated and purified by column chromatography of AB-8 macroporous resin,silica gel,HW-40C,ODS,Sephadex LH-20,and semi-preparative RP-HPLC.Their structures were elucidated by physicochemical properties,spectral analyses and ECD.RESULTS The 70%ethanol extract of Illicium dunnianum was subjected to AB-8 macroporous adsorption resin CC to yield 30%ethanol fraction.Five compounds were obtained and characterized as anisole glycol-7-O-β-L-funanarabifuranosyl-(1→6)-β-D-glucopyranoside(1),4-O-β-D-glucopyranosyloxy-benzaldehyde(2),benzyl-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside(3),β-D-glucopyranoside benzoate(4)and sachalinoside B(5),respectively.CONCLUSION Compound 1 is a new phenolic glycoside,2-5 are identified from Illicium dunnianum for the first time.

Objective:To explore the value of contrast-enhaoced echocardiography for measuring pulmonary transit time（PTT）in assessing heart failure after percutaneous coronary intervention（PCI）in acute ST-segment elevation myocardial infarction（STEMI）patients.Methods:From September 2023 to September 2024，120 patients with STEMI undergoing PCI at Hunan Provincial People's Hospital were prospectively selected and divided into a heart failure group（ n=42）and a non-heart failure group（ n=78）according to the guidelines. The differences in general clinical data，laboratory parameters，and echocardiographic parameters between the two groups were compared. The diagnostic efficacies of PTT，normalized PTT（nPTT），and N-terminal pro-B-type natriuretic peptide（NT-proBNP）were analyzed. Consistency between them and New York Heart Association（NYHA）heart function classification was tested. Results:Compared to the non-heart failure group，the NT-proBNP，PTT，and nPTT values in the heart failure group were significantly increased（all P<0.05）. The area under the curve（AUC）of nPTT was 0.944，better than that of PTT and NT-proBNP（AUC=0.871，0.887）. After K-means clustering reclassified patients into four levels based on nPTT values，nPTT classification showed moderate consistency with NYHA classification（Kappa=0.580， P<0.001），and nPTT differed significantly across NYHA classifications（ P<0.05）. Conclusions:PTT，as an echocardiographic index for assessing cardiac function，has similar diagnostic efficacy to NT-proBNP，the nPTT is even better. It shows moderate consistency with the NYHA classification and holds potential for differentiating overlapping NYHA grades. Importantly，it offers a fresh objective way to evaluate cardiac dysfunction after PCI in STEMI patients.

Aim To investigate the regulatory effects of coenzyme Q10(CoQ10)on blood lipid level and intesti-nal flora abundance in atherosclerotic(As)rats.Methods 24 rats were randomly divided into control group,As group and CoQ10 intervention group.Rats in the As group and CoQ10 intervention group were fed with high-fat chow for 2 weeks,combined with abdominal aortic balloon injury to replicate the As model.CoQ10 was administered by gavage start-ing on the next day of modeling,once daily for 4 weeks.Aortic Movat's staining and lipid levels were used to verify the effect of CoQ10 intervention in As,and the abundance of intestinal flora in intestinal contents was analyzed using met-agenomics.Results Compared with the control group,rat aortic tissues in the As group showed endothelial damage,structural disorganization of the internal elastic plate and inflammatory infiltration,serum triglyceride(TG),total cholester-ol(TC)and low density lipoprotein cholesterol(LDLC)levels were increased,and high density lipoprotein cholesterol(HDLC)levels were decreased.Compared with the As group,the structure of the endothelial cells of the aorta and the structure of the endothelial cells,the internal elastic plates and the smooth muscle cell morphology were relatively regular,serum TC,TG and LDLC levels were decreased,and HDLC levels were increased in the CoQ10 intervention group.Com-pared with the control group,the intestinal bacterial biodiversity in the As group was reduced.At the phylum level,the abundance of the Firmicutes and the Bacteroidetes were down-regulated,whereas that of Proteobacteria was up-regulated.At the genus level,the relative abundance of Lactobacillus,Bacteroides,Akkermansia,Limosilactobacillus,Parabacteroides and Ligilactobacillus was down-regulated,and the relative abundance of Muribaculum was up-regulated.Compared with the As group,CoQ10 intervention restored the biodiversity of the intestinal microbiota in As rats and increased the relative abundance of Firmicutes,Bacteroidetes,Lactobacillus,Bacteroides,Akkermansia,Limosilactobacillus,Parabacteroides and Ligilactobacillus,while reducing the relative abundance of Proteobacteria and Muribaculum(all P＜0.05).Conclusion CoQ10 can regulate blood lipid levels in As rats,upregulate the abundance of beneficial microbiota,downregulate the abun-dance of harmful microbiota,and modulate the diversity of the gut microbiota.

Objective:To study the application effect of the Hazard Analysis and Critical Control Point(HACCP)system in risk assessment and dynamic management of operational safety of medical equipment in surgery rooms.Methods:Based on the HACCP principle,hazard analysis and risk assessment were conducted on the operational safety of equipment in surgery room,and the critical control points were determined,and measures of monitor and management were formulated to conduct whole-process dynamic management for equipment in surgery room.A total of 120 medical equipment in use in the surgery room of Shaanxi Provincial Cancer Hospital from January 2022 to January 2024 were selected.According to random number table method,they were divided into conventional management mode group(60 equipment that received conventional management mode)and dynamic management mode group(60 equipment that received the mode of risk assessment and dynamic management based on HACCP system for operational safety of medical equipment in surgery rooms)to receive different managements.The effect,operational quality,the score of management standardization,and management level for equipment in managing and controlling risk of safety of equipment were compared between two management modes.Results:The average pressure-related injury,electrical injury,extravasation of body fluid,and incidence of unqualified cleaning and disinfection of equipment in the dynamic management mode group were respectively(3.01±1.01)%,(2.65±0.78)%,(1.65±0.64)%and(1.15±0.16)%,all of which were lower than those in the conventional management mode group,and the differences were statistically significant(t=16.311,13.434,12.018,12.629,P<0.05).The average utilization rate,cost-benefit amplification and social benefit amplification of the equipment in the dynamic management mode group were respectively(95.36±2.87)%,(7.98±1.32)%and(10.68±2.36)%,all of which were higher than those in the conventional management mode group,and the average operating rate in the dynamic management mode group was(9.87±1.54)%,which was lower than that in the conventional management mode group,and the differences of the above indicators were significant(t=7.657,15.567,10.354,12.466,P<0.05).The scores of the standard degrees of use and operation,disinfection and sterilization,operation and maintenance,and repairing fault of equipment in the dynamic management mode group were higher than those in the conventional management mode group,and the differences of the above data between two groups were statistically significant(t=16.289,11.415,13.396,9.420,P<0.05),respectively.The rate of standard placement,the rate of reasonable recording and integrity rate of equipment in the dynamic management mode group were higher than those in the conventional management mode group,and the failure rate was lower than that of the conventional management mode group,and the differences of the above indicators between the two groups were statistically significant(x2=6.708,7.070,8.491,13.333,P<0.05).Conclusion:The risk assessment and dynamic management based on HACCP system for operational safety of medical equipment in surgery room can improve the management effect for equipment in surgery room,and reduce the operational risk of equipment,and realize the whole-process management and control for medical equipment in surgery room.

Objective:To screen broadly neutralizing antibodies in human immunodeficiency virus-1（HIV-1）chronically infected individuals and characterize their molecular features and to provide new strategies for rational vaccine development and antibody-based therapeutics.Methods:A total of 34 treatment-na?ve individuals with chronic HIV-1 infection were enrolled. Plasma antibody binding levels were measured against two HIV-1 envelope proteins. Single antigen-specific memory B cells were sorted from high-binding samples，and antibody variable region genes were amplified by PCR for paired expression. The monoclonal antibodies were evaluated for neutralizing activity using pseudovirus assays，and their structural features were analyzed by integrating AlphaFold 3 prediction with Discovery Studio molecular docking.Results:Plasma samples showed strong binding to DU422-GP140 and BG505-GP140. Eight monoclonal antibodies were isolated from two donors. Among them，antibodies 0919-A4，0919-A9 and 0808-A2 could cross-react with GP140 from HIV-1 subtypes AE，BC and B. The monoclonal antibody 0919-A9 demonstrated potent neutralizing activity against SF162（Tier 1）and CH181（Tier 2）pseudoviruses，with somatic hypermutation rates of 13.27%（heavy chain）and 15.58%（light chain）. Structural modeling revealed its specific targeting of the V1V2 region on GP120.Conclusion:The isolated antibody 0919-A9 effectively neutralizes Tier 2 pseudoviruses. Its high somatic mutation frequency and V1V2-targeting property underlie its neutralizing activity，providing both a promising candidate and mechanistic insights for HIV vaccine development and antibody-based therapeutic strategies.

Objective:To evaluate the safety and effectiveness of fuzuloparib for the treatment of ovarian epithelial cancer patients in the real world setting.Methods:A retrospective analysis was conducted on the baseline data of 4 620 ovarian cancer patients who had received fuzuloparib monotherapy or combination therapy. Another 224 ovarian cancer patients who were willing to receive fuzuloparib monotherapy or combination therapy were prospectively enrolled, and their baseline characteristics, drug effectiveness, and safety data were analyzed.Results:(1) Among the 4 620 patients in the retrospective cohort, the median age of patients was 60 years; tumor types: 89.8% (4 149/4 620) had ovarian cancer. Among patients with clearly documented information, the vast majority had a histological type of serous carcinoma (82.9%, 3 770/4 546) and International Federation of Gynecology and Obstetrics (FIGO) staging of Ⅲ-Ⅳ (90.9%, 1 537/1 691). (2) Among the 224 patients in the prospective cohort, the median age of patients was 57 years; tumor types: 83.9% (188/224) had ovarian cancer. Among patients with clearly documented records, the predominant pathologic type was serous carcinoma (91.9%, 193/210), and FIGO stage was Ⅲ-Ⅳ in 79.9% (139/174). (3) Among the 224 prospective patients: 84 patients received first-line fluzoparib maintenance therapy, 92 patients received fluzoparib maintenance therapy after platinum-sensitive recurrence, 23 patients received direct fluzoparib treatment after platinum-sensitive recurrence, 19 patients received direct fluzoparib treatment after platinum-resistant recurrence. The median follow-up durations were 8.5, 8.7, 7.9, and 6.7 months, respectively. The median durations of fluzoparib treatment were 6.7, 4.8, 3.1, and 1.9 months, respectively. The median progression-free survival (PFS) times were not reached during follow-up, 12.6 months, not reached during follow-up, and 4.8 months, respectively. The 1-year PFS rates were 84.1%, 55.0%, 69.8%, and 45.5%, respectively. The remaining 6 patients received other fluzoparib regimens. (4) Among the 224 patients in the prospective dataset, 205 had safety data recorded. Of these, 127 patients (62.0%, 127/205) experienced treatment-related adverse events, with common events including anemia (24.4%, 50/205), thrombocytopenia (21.0%, 43/205), and leukopenia (19.5%, 40/205). Among the 205 patients, 43 (21.0%, 43/205) experienced grade 3 or higher treatment-related adverse events, with common events including anemia (8.3%, 17/205) and thrombocytopenia (8.3%, 17/205).Conclusions:The effectiveness of fuzuloparib in clinical application is generally consistent with other drugs in the same class, with good safety. This study provids new clinical evidence for the treatment of ovarian cancer with fuzuloparib.

Objective:To compare the efficacy of new nasopharyngeal airway and laryngeal mask airway for airway management in the patients undergoing non-intubated video-assisted thoracoscopic surgery (NIVATS).Methods:In this randomised, controlled, non-inferiority trial, 60 American Society of Anesthesiologists Physical Status classification I or Ⅱ patients of both sexes, aged 18-79 yr, scheduled for elective NIVATS from December 2021 to December 2023 at Jining No.1 People′s Hospital, were divided into 2 groups ( n=30 each) using a computer-generated random code in a 1∶1 ratio: new type nasopharyngeal airway group (group N) and laryngeal mask airway group (group L). After anesthesia induction, a new nasopharyngeal airway was inserted in group N, and a laryngeal mask airway was inserted in group L. Spontaneous ventilation was maintained during the NIVATS. Ultrasound-guided serratus anterior plane block was performed on the affected side before anesthesia induction. Anesthesia was maintained with propofol and remifentanil. The primary outcome measure was the rate of intraoperative airway intervention, the airway interventions included repositioning of the airway tools, manual assisted ventilation, jaw-thrust maneuver, and conversion to endotracheal intubation. The secondary outcome measures included the first-attempt success rate of airway device placement, time for establishing a patent airway, the minimum value of SpO 2, the maximum value of P ETCO 2, and incidence of complications such as postoperative sore throat. Results:The rate of intraoperative airway intervention was 27% in group L and 47% in group N ( χ2=2.58, P=0.108). The difference in the rate of intraoperative airway intervention between the two groups was 0.20 (95% confidence interval 0.15-0.25), with a 95% confidence interval upper limit higher than the non-inferiority boundary (10%), indicating that this non-inferiority hypothesis was not established. In comparison to group L, the rate of intraoperative jaw-thrust maneuver intervention was significantly increased, the time to establish a patent airway was shortened, and the incidence of postoperative sore throat was decreased in group N ( P<0.05). Conclusions:Compared with the laryngeal mask airway, the new nasopharyngeal airway can reduce the development of postoperative throat pain, however, it is less effective in maintaining a patent airway. It requires careful consideration of risks and benefits when used for NIVATS.

Objective:To analyze the clinicopathological characteristics and prognostic factors in gastric cancer patients with bone metastasis,and to evaluate the impact of different treatment regimens on survival in patients with synchronous and metachronous bone metastasis.Methods:A total of 120 gastric cancer patients with bone metastasis treated at Nanjing Drum Tower Hospital between 2015 and 2023 were enrolled,including 36 with synchronous bone metastasis and 84 with metachronous bone metastasis.Clinicopathological features were compared between the two groups using the χ2 test.Cox proportional hazards regression model was employed to identify risk factors for overall survival after bone metastasis(OS-BM).The Kaplan-Meier method was used to analyze the effects of different treatments on OS-BM in both synchronous and metachronous groups.Results:Among the 120 patients,104(86.6%)had metastases to other organs.Comparative analysis revealed that synchronous bone metastasis patients exhibited elevated serum C-reactive protein and decreased serum albumin,whereas metachronous bone metastasis patients had reduced peripheral white blood cell and neutrophil counts(all P<0.05).Metachronous bone metastasis(HR=2.35,95%CI[1.47,3.74],P<0.01),serum CA125≥30.2 U/mL(HR=1.6,95%CI[1.03,2.48],P=0.036),white blood cell count≥9.5×10?/L(HR=2.15,95%CI[1.17,3.92],P=0.013),and absence of immunotherapy(HR=2.26,95%CI[1.5,3.39],P<0.01)were independent risk factors affecting patient prognosis.Combined immunotherapy significantly prolonged OS-BM in gastric cancer patients with bone metastasis compared to non-immunotherapy regimens(9.63 vs 4.53 months,P=0.002).Patients with metachronous bone metastasis demonstrated better response to immunotherapy compared to those with synchronous metastases(median OS-BM:10.8 vs 7.3 months,P=0.004).Conclusion:Immunotherapy is an independent protective factor for survival in gastric cancer patients with bone metastasis.Early combination therapy centered on immunotherapy alongside chemotherapy is recommended to prolong survival in such patients.