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*

For Perihilar cholangiocarcinoma (pCCA), surgical resection is the only effective way to cure this disease. However, it has high postoperative mortality and high recurrence rate. Domestic and foreign scholars have constructed statistics-based evaluation methods to predict patients′ postoperative survival and complications, such as nomogram, scoring system and other prognostic models. Based on these methods, clinicians can better select patients who can benefit from surgery and choose the optimal? treatment for more severe patients. Through the adoption of other treatments or some ways to ameliorate some preoperative condition, to improve the patient′s mortality and survival. This article reviews the prognostic risk factors and prognostic models of pCCA in order to provide a reference for clinicians to predict the prognosis about the surgery.

Objective:To establish and validate reference intervals of serum vitamin K for healthy children in China.Methods:A cross-sectional study was conducted from January 2020 to May 2023, involving 807 healthy children aged 0 to 14 years, selected by stratified random sampling based on the population distribution of children in eastern, central, western, and northeastern China. Sample collection was carried out in 16 hospitals across 12 provinces, autonomous regions, and municipalities. Basic information of the children was collected using a standardized self-design questionnaire. Serum levels of vitamin K 1 and vitamin K 2 (menaquinone-4 (MK-4), menaquinone-7 (MK-7)) were measured using liquid chromatography-tandem mass spectrometry. The reference intervals was established by direct approach. The children were divided into different groups by age. Inter-group comparisons were conducted using the Kruskal-Wallis non-parametric test, and the reference intervals ( P2.5- P97.5) were determined using non-parametric methods. Screening 40 healthy children for small sample validation based on age groups within the reference range(25 from eastern, 10 from central, and 5 from western regions). Results:The age of the 807 children was 5.00 (2.00, 9.81) years, and 495 (61.3%) were males and 312 (38.7%) females. Reference intervals were established for 795 children, of whom 303 children were aged 1 month to 3 years and 492 were aged 4 to 14 years. The reference intervals for serum vitamin K 1 were 0.09-4.54 μg/L for children aged 1 month to 3 years, and 0.10-1.73 μg/L for 4-14 years. For MK-7, the intervals were 0.07-1.42 μg/L for 1 month to 3 years and 0.19-2.03 μg/L for 4-14 years. The reference intervals for MK-4 in children aged 1 month to 14 years were 0-0.42 μg/L. The measured values of serum vitamin K 1, MK-4, and MK-7 in the validation samples did not exceed the reference limit in more than 2 samples. Conclusion:Reference intervals for vitamin K 1, MK-4, and MK-7 in healthy children aged 1 month to 14 years have been established and validated, and can be used to assess vitamin K nutritional status in children.

Gallbladder cancer is a common malignant tumor of the biliary tract, characterized by a stealthy onset, high invasiveness, and poor prognosis. The 5-year survival rate of gallbladder cancer patients is only 5%-20%. Currently, radical surgery remains the only potentially curative method for gallbladder cancer. Historically, gallbladder cancer was once considered a contraindication for laparoscopic surgery. In recent years, with the improvement of preoperative diagnosis level of gallbladder cancer, progress in surgical techniques and laparoscopic equipment, an increasing number of clinicians have explored and recognized the effectiveness of laparoscopic surgery for the treatment of gallbladder cancer. This review summarizes the latest advances in laparoscopic surgery for gallbladder cancer.

Objective:To analyze the detection strategy and basic detection situation of markers of infectious diseases transmitted by transfusion in blood testing laboratories of blood stations in China.Methods:Based on the data of practice comparison working party of Blood Stations in Mainland of China from 2017 to 2021, the data on the testing strategies and the basic detection information of the markers for the transmission of infectious diseases through transfusion in the member laboratories of the practice comparison working party of Blood Stations in Mainland of China from 2017 to 2021 were collected, and the situation of the selection for testing markers, testing strategy and the testing method and other relevant aspects were sorted out and analyzed by charts.Results:The selection of the testing markers was consistent, but HTLV testing item was added in some member laboratories. The detection strategy of using two ELISA reagents and one nucleic acid testing (NAT) reagent simultaneously was adopted in 47 member blood stations; 3) NAT method was dominated by mini pool-NAT in member laboratories. The number of members adopting mini-pools of 8 (MP8)-NAT decreased from 17 in 2017 to 14 in 2021, while the number of members adopting mini-pools of 6 (MP6)-NAT increased from 13 in 2017 to 22 in 2021; Roche NAT system accounted for the largest proportion.Conclusions:In order to ensure blood safety and avoid missing detection, the blood stations still adopt the detection strategy of using two ELISA reagents and one nucleic acid testing (NAT) reagent simultaneously; Meanwhile, in order to increase the NAT positive rate, the proportion of mini pool-NAT mainly decreased year by year despite its dominating role, while the proportion of individual donation-NAT increased year by year; NAT method is transiting from mini-pools of 8 (MP8) to mini-pools of 6 (MP6); The proportion of imported NAT system used in NAT laboratory is relatively large.

Objective:To preliminarily estimate and study the effect of nucleic acid testing in blood screening on the residual risk (RR) of transfusion transmitted HBV infection (TTI HBV).Methods:Using the NAT yield/WP ratio model and adopting the relevant data of information management system of practice comparison working party in the Mainland of China, this paper analyzed the trend of the RR of TTI HBV among 18 blood centers from 2015 to 2019 in China, and compared the impact of two kinds of blood screening strategies which were ELISA+ ID-NAT/MP-NAT (individual-donation nucleic acid testing or mini-pool nucleic acid testing) and ELISA + MP-NAT on RR in 2019.Results:The overall trends of the 5-year RR of HBV among 18 blood centers showed by trend chi square test were NAT single positive rate trend χ2= 39.42( P<0.01) and residual risk trend χ2= 279.792( P<0.01); The influence on RR from the differences of ELISA+ ID-NAT/MP-NAT and ELISA+ MP-NAT was statistically significant, and chi square test showed that χ2= 7.4( P<0.01). Conclusions:Since the implementation of nucleic acid testing in the blood screening in China from 2015, the residual risk of transfusion transmitted HBV infection has decreased year by year. The observed two blood screening strategies which dominated in China may lead to discrepancy in the residual risk of TTI.

ObjectiveTo analyze the mental health status and the influencing factors among the residents taking standardized residency training during the outbreak and stable period of COVID-19， so as to provide theoretical basis for their mental health education. MethodsOn February 8 to February 10 and April 11 to April 13， 2020， Symptom Checklist 90 （SCL-90）， Beck Srivastava Stress Inventory （BSSI） and Simplified Coping Style Questionnaire （SCSQ） were distributed in online chat groups involving all grades of residents taking standardized residency training in the Second Affiliated Hospital of Kunming Medical University. Through two rounds of questionnaire survey， non-probability sampling method was used to obtain survey samples， and their mental status were analyzed. ResultsA total of 159 valid questionnaires were collected in the first round， and 99 valid questionnaires in the second round. The first survey showed that the total score of SCL-90 was （117.69±37.74） and the detection rate of positive symptoms in SCL-90 was 25.8%， and mainly dominated by obsession， fear and interpersonal sensitivity. In the second survey， the results showed that the total score of SCL-90 was （127.19±51.44）， and the main positive symptoms included obsession， depression and interpersonal sensitivity， with a positive detection rate of 30.3 %. The first survey found that the mental health status had significant differences among residents of different grades （χ²=7.46， P<0.05）， furthermore， the results indicated that lower grade was a risk factor while non-singleton was the protective factor of mental health status （P<0.05）， and SCL-90 total score was positively correlated with score of negative coping styles （r=0.45， P<0.01）. The second survey also classified lower grade as risk factor and non-singleton as protective factor （P<0.05）， and SCL-90 total score was positively correlated with study stress， economic pressure， interpersonal relationship， clinical practice and negative coping styles （r=0.52， 0.46， 0.55， 0.54， P<0.05 or 0.01）. ConclusionResidents under standardized residency training have obvious mental health problems during the outbreak of COVID-19， and the problems become more serious during the stable period of COVID-19.

OBJECTIVETo explore the role of silent information regulator 1 (SIRT1) signaling pathway in mediating the effect of dexmedetomidine (DEX) to alleviate postoperative cognitive dysfunction (POCD) in aged rats.

METHODSSeventy-two healthy male Sprague-Dawley rats aged 18-20 months (weighing 500-700 g) were randomized equally into normal control group, POCD model group, DEX pretreatment group, and DEX and SIRT1 inhibitor (EX527) pretreatment group. In the latter 2 groups, DEX (25 μg/kg) was injected intraperitoneally in the rats 30 min before the operation, and normal saline was injected instead in the other 2 groups; in EX527 group, EX527 (1 μg/kg) was injected intravenously 5 min before the operation. In all but the control group, the rats were subjected to laparotomy lasting 30 min, and on days 1, 3, and 5 following the operation, 6 rats were randomly selected from each group for Morris water maze test to evaluate their cognitive functions. Immediately after the test, the rats were sacrificed and the hippocampus was collected for determination of the levels of tumor necrosis factor- (TNF-) and interleukin-6 (IL-6) using ELISA; Western blotting was used to detect the expression of SIRT1 and nuclear factor- κB (NF-κB) in the hippocampal neurons.

RESULTSCompared with the control rats, the rats in POCD group and EX527 group showed significantly prolonged escape latency, decreased frequency of crossing the original platform, increased TNF- and IL-6 levels, lowered SIRT1 expression in the hippocampal neurons, and increased NF-κB expression ( < 0.05), and these parameters were comparable between POCD group and EX527 group ( > 0.05). DEX pretreatment significantly alleviated cognitive dysfunction and attenuated the changes in TNF-, IL-6, SIRT1, and NF-κB expressions induced by the operation ( < 0.05), and EX527 pretreatment of the rats obviously blocked the effects of DEX ( < 0.05).

CONCLUSIONSDEX alleviates POCD in aged rats probably via SIRT1 signaling pathway.

Objective To explore the prevalence of depressive disorder in the Mosuo ethnic minority in Ninglang district,Yunnan Province.Methods By stratified random sampling,1 121 Mosuo subjects aged 15 or above were selected and assessed by the MINI-international neuropsychiatric interview according to the Structured Clinical Interview for DSM-V-TR Axis I Disorders-Patient Edition for mental disorders.Results The standardized time-point prevalence of major depressive disorder in Mosuo nationality was 1.74％,and 1.69％ (95 ％ CI =1.32％-2.15 ％) in males and 1.77％ (95 ％ CI =1.39％-2.15 ％) in females.There was no statistically significant difference in the prevalence of major depressive disorder between males and females (x2 =0.051,P＞0.05).The standardized time-point prevalence of dysthymic disorder in Mosuo nationality was 0.78％,and 0.66％ (95％ CI=0.54％-0.78％) in males and 0.88％ (95％ CI=0.74％-1.02％) in females (x2=1.232,P＞0.05).Those aged 40-54 years old had the highest adjusted prevalence of depressive episodes(1.48％ (95％CI=0.77％-2.18％)).Conclusion The prevalence of depressive disorder in Mosuo nationality is in a low level,and the middle-age Mosuo people has the highest time-point prevalence.

Objective To investigate the correlation between normal ascending aortic elasticity and the left ventricular function by 256 slice iCT.Methods 105 subjects who underwent CTA and echocardiography inspection and diagnosed with normal coronary artery and without ascending aortic atherosclerosis were collected.Subjects were divided into three groups according to age and two groups according to gender respectively.Left ventricular function parameter values on CT were measured, including ESV,EDV,SV,EF,CO and MM.The aortic elasticity on CT and echocardiography was measured and calculated respectively on cross-sectional area at 15 mm above aortic valve.The relationship between ascending aortic elasticity and the left ventricular function was evaluated.Results Bland-Altman plot revealed that there was a good conformity between CT and echocardiography in measurement the value of ascending aortic elasticity(ICC=0.988,P<0.05).In different grougs of age,normal ascending aortic elasticity was positively correlated with EDV,ESV,SV,EF and negatively correlated with MM.There was no correlation with the CO.There was a negative relationship between aortic elasticity and age(r=-0.546,P<0.05)