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*

Skeletal muscle injury is a common disease in clinical practice,and an in-depth understanding of its repair mechanisms is crucial for the development of effective therapeutic strategies.This paper focuses on the key role of TGF-β in skeletal muscle injury repair,introduces the diversity of its family members and signaling pathways,explores the expression and regulation part of TGF-β after skeletal muscle injury,analyzes its early expression dynamics and regulatory factors,and thoroughly investigates the effects of TGF-β on skeletal muscle repair,revealing its inflammatory regulation,cellular activation and proliferation as well as fibrosis.Key role.Special attention was paid to its mechanism of action in muscle regeneration and its regulatory mechanism at the cellular level.In addition,the potential clinical applications of TGF-β in the repair of skeletal muscle injury were discussed,and the development and application of it as a therapeutic target and modulator were explored.However,controversies and shortcomings still exist in the current study,such as the dual roles of TGF-β and the impact of individual differences on treatment.Future research directions should include digging deeper into the details of signaling pathways and biomarker discovery.By overcoming these challenges,the potential clinical application of TGF-β in skeletal muscle injury repair is expected to usher in new breakthroughs and provide patients with more individualized and effective treatment strategies.

Objective:To compare the differences of actual absorbed doses of liver malignant tumors after 90Y-selective internal radiation therapy (SIRT) evaluated by 90Y PET/CT and 90Y bremsstrahlung (BRS) SPECT/CT imaging, and to compare the image quality of the 2 imaging methods. Methods:Twenty-one patients (15 males and 6 females; age: (52.4±15.4) years) with liver malignant tumors (15 cases of primary liver cancer, 6 cases of liver metastases; 39 lesions) between September 2021 and August 2022 were retrospectively analyzed. All patients underwent both 90Y PET/CT imaging and 90Y BRS SPECT/CT imaging in the Department of Nuclear Medicine, Hainan Cancer Hospital. The ratios of the actual absorbed doses based on 90Y PET/CT imaging and 90Y BRS SPECT/CT imaging to the lowest standard absorbed dose(100 Gy) for tumor response were calculated. The image contrast and distinguishability of the two imaging methods were scored. Wilcoxon signed rank test and Wilcoxon rank sum test were used for data analysis. Results:The tumor absorbed doses evaluated by 90Y PET/CT and 90Y BRS SPECT/CT were 143.94(55.91, 233.48) Gy and 107.82(53.59, 157.53) Gy respectively. The doses evaluated by 90Y PET/CT were higher than the standard threshold in 24 lesions, while 19 lesions showed higher evaluated doses by 90Y BRS SPECT/CT than the standard threshold. Compared with 90Y PET/CT, 90Y BRS SPECT/CT underestimated the tumor absorbed dose of -24.25%(-32.32%, -12.14%). The ratio of dose evaluated by 90Y PET/CT to the lowest standard threshold was 1.33(0.56, 1.91), which was higher than that of dose evaluated by 90Y BRS SPECT/CT to the lowest standard threshold (0.97(0.47, 1.25); z=0.04, P<0.001). PET/CT image contrast was scored 0, 1, 2, 3 in 2, 2, 12, 23 lesions respectively, and SPECT/CT image contrast was scored 0, 1, 2, 3 in 2, 3, 16, 18 lesions respectively ( z=-1.29, P=0.199). The distinguishability scores of 0, 1, 2 based on PET/CT images were found in 3, 15, 21 lesions, while those based on SPECT/CT images were found in 4, 32, 3 lesions respectively ( z=-2.79, P=0.005). Conclusion:90Y PET/CT imaging is superior to 90Y BRS SPECT/CT imaging in radiation dose evaluation and tumor focus differentiation in patients with liver malignant tumors after 90Y-SIRT.

Objective:To investigate the clinicopathological features and differential diagnosis of eosinophilic vacuolated tumor (EVT).Methods:Seven cases of EVT with characteristic morphology and unequivocal diagnosis from the Affiliated Hospital of Qingdao University (6 cases), Qingdao, China and the 971 Hospital of PLA Navy (1 case), Qingdao, China between January 2010 and December 2021 were subject to morphological and immunohistochemical analyses. Additionally, whole exome sequencing (WES) was performed in two cases. Twenty-two cases of renal oncocytoma (RO) and 17 cases of eosinophilic chromophobe renal cell carcinoma (eChRCC) diagnosed at the same time were used as controls.Results:Four males and three females with a mean age of 42 years (range: 29-61 years) were included in the study. The tumors were nodular and well-circumscribed, with sizes ranging from 1.5 to 4.5 cm. On cross-section, they appeared gray-red or gray-white, solid, and soft. Tumor cells were arranged in nests, solid sheets, and acinar or small vesicular structures. These cells exhibited eosinophilic cytoplasm with large, prominent clear vacuoles and round nuclei with prominent nucleoli. Perinuclear halos were focally present in four cases, while small tumor cells with sparse cytoplasm and hyperchromatic nuclei were seen in one case. No necrosis or mitosis was noted. Edematous stroma was detected in three cases. All tumors were positive for CD117 and Cathepsin K, but negative for vimentin and CK7. CK20 was positive in scattered individual cells, and Ki-67 positivity ranged from 1% to 4%. Point mutations in MTOR were identified in both patients who were subject to the molecular analysis. Statistical differences in the expression of Cathepsin K, CD10, S-100A1, and Cyclin D1 between EVT and RO ( P<0.05) were significant, so were the differences in the expression of Cathepsin K, CD10, CK7 and claudin 7 between EVT and eChRCC ( P<0.001). Seven patients were followed up for 4 to 96 months (mean, 50 months), with no recurrences or metastases. Conclusions:EVT is a rare renal tumor that shares morphological and immunophenotypic features with RO and eChRCC, and it is closely linked to the TSC/MTOR pathway. The presence of large prominent transparent vacuoles in eosinophilic cytoplasm along with conspicuous nucleoli is its key morphological characteristics. The use of combined immunohistochemical stains greatly aids in its diagnosis. Typically, the tumor exhibits indolent biological behaviors with a favorable prognosis.

Objective:To investigate the application of virtual reality (VR) technology on intraoperative pain in patients undergoing peritoneal dialysis (PD)-related procedures with local infiltration anesthesia and the satisfaction.Methods:It was a single-center, prospective, concurrent controlled study. Patients were divided into two groups: VR group and control group. In the VR group, patients wore a VR headset to watch soothing audio and video content during surgery, while the control group underwent routine procedures. Intraoperative pain and satisfaction were assessed using the visual analog scale (VAS) and a 5-point satisfaction scale within 30 minutes of surgery. In addition, tolerance of the VR experience in the VR group was assessed using the VR sickness questionnaire.Results:A total of 43 patients were included in the study, including 25 males (58.1%). Chronic glomerulonephritis [17 cases (39.5%)] and diabetic nephropathy [6 cases (14.0%)] were the main primary diseases. There were 23 cases in the control group and 20 cases in the VR group. There were no significant differences between the two groups in age, sex ratio, proportion of primary disease, diabetes, hypertension, distribution of operation methods, preoperative vital signs and operation time (all P>0.05). VAS pain score was significantly lower in the VR group than that in the control group (5.90±2.38 vs. 7.43±1.67, t=2.469, P=0.018). The percentage of patients who were satisfied was 89.5% (17/19) in the VR group and 78.3% (18/23) in the control group, but there was no significant difference (chi-square test for continuity correction, χ2=0.308, P=0.579). Three patients in the VR group withdrew from the study due to severe discomfort, while the remaining participants found the VR experience to be tolerable. Common adverse effects included fatigue and blurred vision. Conclusions:The application of VR technology in PD-related procedures has been effective in reducing intraoperative pain when combined with local infiltration anesthesia. Furthermore, the utilization of VR technology in PD-related procedures is associated with a safe and tolerable outcome, despite the observation of some adverse effects.

OBJECTIVE: To investigate the development present situation of the department of critical care medicine in Inner Mongolia Autonomous Region (hereinafter referred to as Inner Mongolia), in order to promote the standardized and homogeneous development of critical care medicine in Inner Mongolia, and also provide a reference for discipline construction and resource allocation. METHODS: A survey study was conducted in comprehensive intensive care unit (ICU) of tertiary and secondary hospitals in Inner Mongolia by online questionnaire survey and telephone data verification. The questionnaire was based on the Guidelines for the Construction and Management of Intensive Care Units (Trial) (hereinafter referred to as the Guidelines) issued by the National Health Commission in 2009 and the development trend of the discipline. The questionnaire covered six aspects, including hospital basic information, ICU basic information, personnel allocation, medical quality management, technical skill and equipment configuration. The questionnaire was distributed in September 2022, and it was filled out by the discipline leaders or department heads of each hospital. RESULTS: As of October 24, 2022, a total of 101 questionnaires had been distributed, 85 questionnaires had been recovered, and the questionnaire recovery rate had reached 84.16%, of which 71 valid questionnaires had been collected in a total of 71 comprehensive ICU. (1) There were noticeable regional differences in the distribution of comprehensive ICU in Inner Mongolia, with a relatively weak distribution in the east and west, and the overall distribution was uneven. The development of critical care medicine in Inner Mongolia was still lacking. (2) Basic information of hospitals: the population and economy restricted the development of ICU. The average number of comprehensive ICU beds in the western region was only half of that in the central region (beds: 39.0 vs. 86.0), and the average number of ICU beds in the eastern region was in the middle (83.6 beds), which was relatively uneven. (3) Basic information of ICU: among the 71 comprehensive ICU surveyed, there were 44 tertiary hospitals and 27 secondary hospitals. The ratio of ICU beds to total beds in tertiary hospitals was significantly lower than that in secondary hospitals [(1.59±0.81)% vs. (2.11±1.07)%, P < 0.05], which were significantly lower than the requirements of the Guidelines of 2%-8%. The utilization rate of ICU in tertiary and secondary hospitals [(63.63±22.40)% and (44.65±20.66)%, P < 0.01] were both lower than the bed utilization rate required by the Guidelines (75% should be appropriate). (4) Staffing of ICU: there were 376 doctors and 1 117 nurses in tertiary hospitals, while secondary hospitals had 122 doctors and 331 nurses. There were significant differences in the composition ratio of the titles of doctors, the degree of doctors, and the titles of nurses between tertiary and secondary hospitals (all P < 0.05). Most of the doctors in tertiary hospitals had intermediate titles (attending physicians accounted for 41.49%), while most of the doctors in secondary hospitals had junior titles (resident physicians accounted for 43.44%). The education level of doctors in tertiary hospitals was generally higher than that in secondary hospitals (doctors: 2.13% vs. 0, masters: 37.24% vs. 8.20%). The proportion of nurses in tertiary hospitals was significantly lower than that in secondary hospitals (17.01% vs. 24.47%). The ratio of ICU doctors/ICU beds [(0.64±0.27)%, (0.59±0.34)%] and ICU nurses/ICU beds [(1.76±0.56)%, (1.51±0.48)%] in tertiary and secondary hospitals all failed to meet the requirements above 0.8 : 1 and 3 : 1 of the Guidelines. (5) Medical quality management of ICU: compared with secondary hospitals, the proportion of one-to-one drug-resistant bacteria care in tertiary hospitals (65.91% vs. 40.74%), multimodal analgesia and sedation (90.91% vs. 66.67%), and personal digital assistant (PDA) barcode scanning (43.18% vs. 14.81%) were significantly higher (all P < 0.05). (6) Technical skills of ICU: in terms of technical skills, the proportion of bronchoscopy, blood purification, jejunal nutrition tube placement and bedside ultrasound projects carried out in tertiary hospitals were higher than those in secondary hospitals (84.09% vs. 48.15%, 88.64% vs. 48.15%, 61.36% vs. 55.56%, 88.64% vs. 70.37%, all P < 0.05). Among them, the placement of jejunal nutrition tube, bedside ultrasound and extracorporeal membrane oxygenation were mainly completed independently in tertiary hospitals, while those in secondary hospitals tended to be completed in cooperation. (7) Equipment configuration of ICU: in terms of basic equipment, the ratio of the total number of ventilators/ICU beds in tertiary and secondary hospitals [0.77% (0.53%, 1.07%), 0.88% (0.63%, 1.38%)], and the ratio of injection pump/ICU beds [1.70% (1.00%, 2.56%), 1.25% (0.75%, 1.88%)] didn't meet the requirements of the Guidelines. The equipment ratio was insuffcient, which means that the basic needs of development had not been met yet. CONCLUSIONS The development of comprehensive ICU in Inner Mongolia has tended to mature, but there is still a certain gap in the development scale, personnel ratio and instruments and equipment compared with the Guidelines. Moreover, the comprehensive ICU appears the characteristics of relatively weak eastern and western regions, and the overall distribution is uneven. Therefore, it is necessary to increase efforts to invest in the construction of the department of critical care medicine.
Humans ; Intensive Care Units ; Critical Care ; Surveys and Questionnaires ; Tertiary Care Centers ; China

[This corrects the article DOI: 10.1016/j.apsb.2022.06.009.].

Objective To evaluate the impact of customized rehabilitation exercise plans based on the results of cardiopulmonary exercise test(CPET)on cardiac function and prognosis in patients with chronic heart failure(CHF).Methods A total of 52 CHF patients admitted to Chengde Central Hospital from February 2020 to September 2021 were selected as the study subjects,and the patients were divided into observation group and control group according to the principle of randomized controlled study,with 26 cases in each group.The control group received rehabilitation treatment excluding exercise.The observation group was given routine rehabilitation treatment and high-intensity rehabilitation exercise plans based on CPET guidance.Above anaerobic threshold Δ50%power was exercise intensity,exercise time was 30 minutes/day,4 days/week,and intervention period was 12 weeks.Before and 12 weeks after intervention,CPET functional indicators,serum brain natriuretic peptide(BNP),left ventricular ejection fraction(LVEF),left ventricular end-diastolic diameter(LVEDD),and 6-minute walking distance(6MWD)were measured.The Minnesota CHF quality of life questionnaire(LiHFe)was used to evaluate patient's quality of life,readmission rate and cardiogenic mortality within 1 year of follow-up,and univariate and multivariate Logistic regression analysis was used to analyze factors affecting readmission of CHF patients.Results ① Cardiopulmonary function indicators:there was no statistically significant difference in the anaerobic threshold,peak oxygen uptake,and peak oxygen pulse of CPET functional indicators before and after intervention in the control group,after intervention,the CPET functional indicators in the observation group were significantly higher than those before intervention,and the above indexes in the observation group were significantly higher than those of the control group[anaerobic threshold(mL·min-1·kg-1):10.77±1.40 vs.9.59±1.11,anaerobic threshold(%ped):78.95±11.39 vs.70.09±6.48,peak oxygen uptake(mL·min-1·kg-1):15.63±1.36 vs.14.27±1.72,peak oxygen uptake(%ped):72.42±6.91 vs.63.41±7.31,peak oxygen pulse(mL/order):11.38±1.29 vs.9.05±1.64,peak oxygen pulse(%ped):90.23±10.16 vs.80.53±6.73,all P＜0.05].②Serum indicators,cardiac function indicators,exercise ability indicators,and quality of life evaluation:there was no statistically significant difference in serum indicators BNP,cardiac function indicators LVEDD,LVEF,exercise ability indicators 6MWD,and quality of life LiHFe scores between the two groups before intervention.After intervention,BNP and LiHFe scores were significantly reduced compared with before intervention,while LVEF and 6MWD were both increased compared with before intervention,and the changes of the above indexes in the observation group were more significant than those in the control group[BNP(ng/L):313.25±77.91 vs.445.89±110.67,LVEF:0.41±0.08 vs.0.37±0.06,6MWD(m):495.62±91.35 vs.416.04±65.29,LiHFe score:23.27±6.02 vs.29.50±4.61,all P＜0.05].③ Prognostic follow-up:the readmission rate within 1 year in the observation group was significantly lower than that in the control group(23.08%vs.53.85%),and there was no statistically significant difference in the mortality rate of cardiogenic diseases between the two groups.④Logistic univariate analysis showed that hyperlipidemia,New York Heart Association(NYHA)grading,BNP,and rehabilitation exercise were factors that affect the prognosis of CHF patients[odds ratio(OR)and 95%confidence interval(95%CI)were 0.098(0.019-0.494),0.069(0.016-0.294),1.018(1.007-1.029),and 3.889(1.178-12.841),respectively,all P＜0.05].Multivariate analysis showed that after adjusting for other factors,hyperlipidemia,NYHA grading,and BNP were risk factors affecting the prognosis of CHF patients(OR and 95%CI were 0.068(0.007-0.687),0.048(0.005-0.415),1.016(1.002-1.030),respectively,with P＜0.05],the use of rehabilitation exercise therapy was a protective factor affecting the prognosis of CHF patients[OR and 95%CI were 11.179(1.135-10.124),P＜0.05].Receiver operator characteristic curve(ROC curve)analysis showed that hyperlipidemia,NYHA grading,BNP,rehabilitation exercise therapy,and combined testing all had predictive value for the patient's prognosis(all P＜0.05),and the prediction value of joint detection was the highest,with the area under the ROC curve(AUC)=0.984 and P = 0.000.Conclusion Developing a high-intensity individualized cardiac exercise rehabilitation plan under the guidance of CPET can help improve the cardiopulmonary function,cardiac function,and quality of life of CHF patients,which is of great benefit for improving the long-term prognosis of CHF patients and has high safety.

MAGED4B belongs to the melanoma-associated antigen family; originally found in melanoma, it is expressed in various types of cancer, and is especially enriched in glioblastoma. However, the functional role and molecular mechanisms of MAGED4B in glioma are still unclear. In this study, we found that the MAGED4B level was higher in glioma tissue than that in non-cancer tissue, and the level was positively correlated with glioma grade, tumor diameter, Ki-67 level, and patient age. The patients with higher levels had a worse prognosis than those with lower MAGED4B levels. In glioma cells, MAGED4B overexpression promoted proliferation, invasion, and migration, as well as decreasing apoptosis and the chemosensitivity to cisplatin and temozolomide. On the contrary, MAGED4B knockdown in glioma cells inhibited proliferation, invasion, and migration, as well as increasing apoptosis and the chemosensitivity to cisplatin and temozolomide. MAGED4B knockdown also inhibited the growth of gliomas implanted into the rat brain. The interaction between MAGED4B and tripartite motif-containing 27 (TRIM27) in glioma cells was detected by co-immunoprecipitation assay, which showed that MAGED4B was co-localized with TRIM27. In addition, MAGED4B overexpression down-regulated the TRIM27 protein level, and this was blocked by carbobenzoxyl-L-leucyl-L-leucyl-L-leucine (MG132), an inhibitor of the proteasome. On the contrary, MAGED4B knockdown up-regulated the TRIM27 level. Furthermore, MAGED4B overexpression increased TRIM27 ubiquitination in the presence of MG132. Accordingly, MAGED4B down-regulated the protein levels of genes downstream of ubiquitin-specific protease 7 (USP7) involved in the tumor necrosis factor-alpha (TNF-α)-induced apoptotic pathway. These findings indicate that MAGED4B promotes glioma growth via a TRIM27/USP7/receptor-interacting serine/threonine-protein kinase 1 (RIP1)-dependent TNF-α-induced apoptotic pathway, which suggests that MAGED4B is a potential target for glioma diagnosis and treatment.
Humans ; Tumor Necrosis Factor-alpha ; DNA-Binding Proteins/metabolism* ; Ubiquitin-Specific Peptidase 7 ; Cisplatin ; Temozolomide ; Transcription Factors ; Glioma ; Cell Proliferation ; Melanoma ; Cell Line, Tumor ; Apoptosis ; Nuclear Proteins/genetics*