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*

In this comprehensive review, we delve into the evolution of drug delivery systems in reproductive medicine with a focus on the emerging role of exosomes, a class of extracellular vesicles. Exosomes offer unique advantages in overcoming these challenges due to their inherent biocompatibility, stability, and ability to facilitate targeted delivery. This review provides a detailed examination of exosome biogenesis and their function in cellular communication, setting the stage for understanding their potential as drug delivery vehicles. We explore the mechanisms through which exosomes can be loaded with small molecule drugs and the benefits they offer over synthetic nanoparticles. The review highlights groundbreaking case studies that illustrate the successful application of exosome-mediated drug delivery in reproductive health, including enhancing fertility treatments, supporting gamete and embryo development, and facilitating maternal-fetal communication. This study aims to provide a precise understanding of how exosomal drug delivery can revolutionize treatments for reproductive health disorders, paving the way for future therapeutic applications. Lastly, we touch upon the promising therapeutic implications of exosomal delivery for proteins and genes, offering a window into future treatments for reproductive health disorders.

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 investigate the difference in intracranial infection rate between long-tunneled external ventricular drainage(LTEVD)and conventional external ventricular drainage(EVD),as well as the risk factors for intracranial infection.Methods:A retro-spective analysis was performed for the clinical data of 45 patients who were admitted to Department of Neurology Center,The Third Affiliated Hospital of Chongqing Medical University,from January 2020 to December 2022 and underwent EVD,among whom 13 patients underwent LTEVD(LTEVD group)and 32 patients underwent conventional EVD(EVD group).Related data were recorded for both groups,including general information,postoperative catheter-related complications,and postoperative management,to investi-gate the effect on reducing the rate of intracranial infection.According to the presence or absence of intracranial infection after surgery,the patients were divided into the infection group with 10 patients and non-infection group with 35 patients,and related clini-cal data were analyzed to investigate the risk factors for intracranial infection.Results:The LTEVD group had a significantly lower secondary infection rate of catheterization days than the EVD group[2.40‰(1/417)vs.27.19‰(9/331),P=0.009].The duration of catheterization was 14-85 days[27.00(22.50,36.50)days]in the LTEVD group and 8-22 days[9.00(8.00,11.50)days]in the EVD group,suggesting that the LTEVD group had a significantly longer duration of catheterization than the EVD group(P=0.000).The multivariate logistic regression analysis showed that the times of cerebrospinal fluid sampling was an independent risk factor for post-operative intracranial infection in patients undergoing EVD,and the use of LTEVD was a protective factor against intracranial infection after EVD.Conclusion:Compared with conventional EVD,LTEVD can safely prolong the duration of catheterization and reduce the rate of postoperative intracranial infection in patients undergoing EVD.The use of LTEVD procedure and the reduction in the times of cerebrospinal fluid sampling can reduce the risk of postoperative in-tracranial infection.

Objective To explore the interaction between long non-coding RNA growth arrest-specific 5(GAS5)and miR-135b-5p/adenomatous polyposis coli(APC)axis,and to elucidate its biological function in the proliferation,metastasis and invasion of glioma.Methods The relationship of GAS5 expression level with survival rate of glioma patients was analyzed based on CGGA database.qRT-PCR was used to detect the expression level of GAS5 in glioma tissues and cell lines.Human glioma T98 and A172 cell lines were subjected for overexpression and knockdown of GAS5,miR-135b-5p and APC by transfection.Western blotting and qRT-PCR were applied to measure the expression of related genes in glioma tissues or cell lines.CCK-8,Transwell and wound healing assays was conducted to determine the effect of GAS5 on the viability and motility of glioma cells.Moreover,dual luciferase reporter analysis were utilized to elucidate the regulatory mechanisms of GAS5 and miR-135b-5p/APC.Results Analysis on CGGA database showed that the survival rate of glioma patients with low GAS5 expression was significantly decreased(P＜0.001).Western blotting and qRT-PCR indicated that GAS5 was down-regulated in the glioma cell lines and tissues than human normal astrocytes and para-cancer tissues(P＜0.01).The results of CCK-8,Transwell and wound healing assays revealed that overexpression of GAS5 significantly inhibited the viability,invasion and migration abilities in T98 cells when compared with the cells of the vector group(P＜0.01).Dual luciferase reporter analysis displayed that there were binding sites between GAS5 and miR-135b-5p,and the expression of the two was negatively correlated(P=0.019).Further studies suggested that the effect of GAS5 on biological function in glioma cells was through targeting miR-135b-5p,and overexpression of APC reversed the promoting effect of miR-135b-5p on glioma cells(P＜0.01).Western blotting displayed that enhancing miR-135b-5p expression inhibited the expression of APC(P＜0.001),and GAS5 upregulated the expression of APC by targeting miR-135b-5p(P＜0.01).Conclusion GAS5 inhibits the progression of glioma via miR-135b-5p/APC axis,which revealing the molecular regulatory mechanism of GAS5/miR-135b-5p/APC.

Objective To investigate the therapeutic effects and underlying mechanisms of rutaecarpine on Helicobacter pylori(Hp)-induced chronic atrophic gastritis(CAG).Methods An Hp-induced CAG rat model was established.Successfully modeled rats were randomly divided into model group,low-dose rutaecarpine group,medium-dose rutaecarpine group,and high-dose rutaecarpine group,with 12 rats in each group.An additional 12 rats served as the normal control group.Body mass changes were recorded before and after treatment.Gastric mucosal histopathology was analyzed using hematoxylin-eosin(HE)staining.Levels of inflammatory cytokines(TNF-α,IL-6,IL-10)in gastric mucosal supernatants were measured by enzyme-linked immunosorbent assay(ELISA).mRNA expression levels of inducible nitric oxide synthase(iNOS),cluster of differentiation 86(CD86),arginase 1(Arg-1),and mannose receptor(CD206)in gastric mucosal tissues were detected by real-time quantitative polymerase chain reaction(RT-qPCR).Protein expression levels of nuclear factor κB(NF-κB)pathway-related proteins were determined by Western Blot.Results Compared with the normal group,the model group exhibited disorganized gastric mucosal epithelium,reduced glandular structures in the lamina propria,significant inflammatory cell infiltration,and elevated gastric mucosal histopathology scores.TNF-α,IL-6,and IL-10 levels in gastric mucosal supernatants,iNOS and CD86 mRNA expression,and phosphorylated NF-κB inhibitor α(p-IκBα)and phosphorylated NF-κB p65 subunit(p-p65)protein levels were significantly increased,while Arg-1 and CD206 mRNA expression were significantly decreased,the difference being statistically significant(P＜0.05).Compared with the model group,medium-and high-dose rutaecarpine treatment reduced inflammatory cell infiltration,restored cellular arrangement,increased glandular structures in the lamina propria,and significantly lowered gastric mucosal histopathology scores,TNF-α,IL-6,and IL-10 levels,iNOS and CD86 mRNA expression,and p-p65 and p-IκBα protein expression were significantly reduced,whereas Arg-1 and CD206 mRNA expression were significantly increased,the difference being statistically significant(P＜0.05),with dose-dependent effects.Conclusion Rutaecarpine ameliorates Hp-induced CAG by modulating macrophage polarization and attenuating inflammatory responses,likely through downregulation of p-p65 and p-IκBα expression and subsequent inhibition of NF-κB pathway activation.

ObjectiveTo explore the therapeutic effect and mechanism of Guipitang on rats with myocardial ischemia. MethodFifty SD rats were divided into five groups： a control group， a model group， low and high-dose Guipitang （7.52， 15.04 g·kg^-1） groups， and a trimetazidine group （0.002 g·kg^-1）. By intragastric administration of vitamin D₃ and feeding rats with high-fat forage and injecting isoproterenol， the rat model of myocardial ischemia was established. After drug treatment of 15 d， an electrocardiogram （ECG） was performed to analyze the degree of myocardial injury. A fully automatic biochemical analyzer was used to detect the changes in the serum levels of total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C）. Hematoxylin-eosin （HE） staining and Masson staining were used to observe myocardial histopathological changes. TdT-mediated dUTP nick end labeling （TUNEL） staining was used to detect cardiomyocyte apoptosis. Western blot was adopted to detect the protein levels of extracellular signal-regulated kinase 1/2 （ERK1/2）， phospho-ERK1/2 （p-ERK1/2）， p38 mitogen-activated protein kinase （p38 MAPK）， phospho-p38 MAPK （p-p38 MAPK）， B-cell lymphoma-2 （Bcl-2）-associated X （Bax）， Bcl-2， and cleaved cysteine aspartate proteolytic enzyme （cleaved Caspase-3）. ResultCompared with the control group， the ECG S-T segment decreased in the model group. The serum levels of TC， TG， and LDL-C were increased significantly （P<0.05）. The arrangement of myocardial tissue was disordered， and the proportion of cardiomyocyte apoptosis increased. The protein levels of cleaved Caspase-3， Bax， and p-p38 MAPK in the heart were increased， and the Bcl-2 expression was decreased （P<0.05）. Compared with the model group， the S-T segment downward shift was restored in the low and high-dose Guipitang groups and trimetazidine group， and the levels of TC， TG， and LDL-C were decreased. The protein expression of cleaved Caspase-3 and Bax in the heart dropped， and p-p38 MAPK and p-ERK1/2 protein expressions increased significantly （P<0.05）. The degree of myocardial injury was alleviated， and the proportion of cardiomyocyte apoptosis decreased. Bcl-2 protein expression was increased significantly in the low-dose Guipitang group （P<0.05）. ERK1/2 and p38 MAPK proteins had no significant difference among different groups. ConclusionGuipitang could alleviate myocardial injury and inhibit cardiomyocyte apoptosis in rats by activating the expression of ERK1/2 and p38 MAPK.

Objective To study the application value of 3.0T MRI T2 mapping quantitative technology in the diagnosis of anterior cruciate ligament sprain and chronic injury of knee joint.Methods A total of 82 subjects were studied,and the experimental group 72 cases was divided into grade Ⅰ injury group(25 cases),grade Ⅱ injury group(25 cases),chronic injury group(22 cases),and control group 10 cases.The experimental group met the criteria of arthroscopy.The proximal,middle,and distal segments of the anterior cruciate ligament were selected as the region of interest(ROI),and T2 mapping values were measured.The differences in T2 mapping values of each area were compared between and within the groups,while compared with arthroscopy.Results The T2 mapping values in grade Ⅰ,Ⅱ,and chronic injury groups were higher than those in control group(P<0.05).Comparison within the experimental group:the T2 mapping values of each area in grade Ⅱ injury group were higher than those in grade Ⅰ injury group and chronic injury group(P<0.05).The T2 mapping values of each area in grade Ⅰ injury group were higher than those in chronic injury group(P<0.05).The specificity,sensitivity,positive predictive value,negative predictive value and accuracy of T2 mapping in diagnosing anterior cruciate ligament grade Ⅰ injury were 94.7%,95.5%,89.7%,96.6%,and 90.2%respectively.The specificity,sensitivity,positive predictive value,negative predictive value,and accuracy of grade Ⅱ injury were 89.4%,87.9%,92.1%,93.4%,and 93.8%respectively.The specificity,sensitivity,positive predictive value,negative predictive value,and accuracy of chronic injury were 92.2%,95.4%,90.3%,87.6%,and 91.5%respectively.Kappa test showed a good con-sistency between T2 mapping results and arthroscopic results,with a Kappa value of 0.763(P<0.01).Conclusion The value of MRI T2 mapping can provide a reference for the clinical diagnosis of anterior cruciate ligament sprain and chronic injury of knee joint,and the results are in good agreement with the control of arthroscopy.

Objective:To explore the effectiveness of a hysteroscopic endometrial lesion diagnosis model de-veloped based on deep learning(DL)algorithm combined with gradient-weighted class activation mapping(Grad-CAM)visualization technology.Methods:303 hysteroscopy videos(4781 images)of 291 patients who un-derwent hysteroscopy examination in the Department of Gynecology,Renmin Hospital of Wuhan University from June 1,2021 to December 31,2022 were selected.The dataset was divided into a training set(3703 images)and a test set(1078 images)by weight sampling method.After the training set was used for model learning and train-ing,two model architectures,residual neural network(ResNet18)and efficient neural network(EfficientNet-B0),were selected to verify the model in the test set by five-class and two-class classification tasks,respectively.Tak-ing histopathology as the gold standard,the diagnostic efficacy was evaluated to select the optimal model,and the Grad-CAM layer was embedded in the optimal model to output hysteroscopy images of Grad-CAM.Results:①In the five-class classification tasks,the accuracy of EfficientNet-B0 model(93.23％)was higher than that of Res-Net18 model(84.23％);the area under the curve(AUC)of EfficientNet-B0 model in the diagnosis of five disea-ses,including atypical endometrial hyperplasia,endometrial polyps,endometrial cancer,endometrial atypical hy-perplasia,and submucous myoma,was slightly higher than that of ResNet18 model,and the AUC of both models was almost above 0.980.②In the binary classification task of accuracy and the evaluation of specificity,the two models were similar,both above 93.00％,and the sensitivity of EfficientNet-B0 model(91.14％)was significantly better than that of ResNet18 model(77.22％).③EfficientNet-B0 model combined with Grad-CAM algorithm could identify the abnormal areas in the image.After biopsy and pathological examination,it was confirmed that about 95％of the marked areas in the model's output heatmap were lesion areas.Conclusions:The hysteroscopy di-agnostic model developed by EfficientNet-B0 model combined with Grad-CAM has high diagnostic accuracy,sen-sitivity,and specificity,and has application value in the diagnosis of endometrial lesions.