Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Naringenin (4,5,7-trihydroxyflavonoid) is a naturally occurring bioflavonoid found in citrus fruits, which plays an important role in metabolic syndrome, neurological disorders, and cardiovascular diseases. However, the pharmacological mechanism and biological function of naringenin on anti-angiogenesis and anti-tumor immunity have not yet been elucidated. Our study firstly demonstrates that naringenin inhibits the growth of hepatocellular carcinoma (HCC) cells both in vivo and in vitro. Naringenin diminishes the ability of HCC cells to induce tube formation and migration of human umbilical vein endothelial cells (HUVECs) and suppresses neovascularization in chicken chorioallantoic membrane (CAM) assays. Meanwhile, in vivo results demonstrate that naringenin can significantly upregulate level of CD8⁺ T cells, subsequently increasing the level of immune-related cytokines in the tumor immune microenvironment. Mechanistically, we found that naringenin facilitate the K48-linked ubiquitination and subsequent protein degradation of vascular endothelial growth factor A (VEGFA) and mesenchymal-epithelial transition factor (c-Met), which reduces the expression of programmed death ligand 1 (PD-L1). Importantly, combination therapy naringenin with PD-L1 antibody or bevacizumab provided better therapeutic effects in liver cancer. Our study reveals that naringenin can effectively inhibit angiogenesis and anti-tumor immunity in liver cancer by degradation of VEGFA and c-Met in a K48-linked ubiquitination manner. This work enlightens the potential effect of naringenin as a promising therapeutic strategy against anti-angiogenesis and anti-tumor immunity in HCC.

OBJECTIVE This study aimed to investigate the effects of Buyang Huanwu Decoction(BYHWD)on delaying vascu-lar aging and explore whether the underlying mechanism is associated with microRNA-665(miR-665)/DNA damage-regulated autoph-agy modulator1(DRAM1)-mediated autophagy.METHODS Male SD rats with natural aging were randomly divided into the aging group,BYHWD low,medium,high dosage groups(9.25,18.5,37.0 g·kg-1)and resveratrol group(80 mg·kg-1),with a young group set as well.The rats in each group were dissected and the blood vessels were collected.ELISA was used to assess senescence as-sociated β-galactosidase(SA-β-Gal)activity and advanced glycation end products(AGEs)level in vascular tissues.HE,Masson,and EVG staining were performed to observe the morphological structure of the vascular tissues.The qPCR was performed to detect the expression of miR-665 in vascular tissues.Bioinformatics analysis and dual-luciferase reporter gene experiments were used to validate the targeting relationship between miR-665 and DRAM1.Transmission electron microscope was used to observe the autophagosome.Western blot was performed to determine the protein expression of p16,DRAM1 and autophagy-related proteins Bec-lin1,p62 and LC3.Immunohistochemistry was used to detect the protein expression of DRAM1 in vascular tissues.RESULTS Com-pared to the young group,the aging group showed increased SA-β-Gal activity,AGEs level and p16 protein expression(P<0.01),disordered arrangement of vascular tissues,thickened media,increased collagen fibers,fractured and disorganized elastic fibers.The expression of miR-665 was upregulated(P<0.01).The number of autophagosomes was reduced.The protein expression of Beclin1 and LC3Ⅱ/Ⅰ downregulated(P<0.01),while the protein expression of p62 was upregulated(P<0.01).In addition,the protein ex-pression of DRAM1 was downregulated in vascular tissues(P<0.01).Compared to the aging group,intervention with BYHWD and resveratrol reduced SA-β-Gal activity(P<0.01),AGEs level and p16 protein expression(P<0.05,P<0.01),improved vascular morphology and elastic fiber structure,reduced collagen fibers.High dose BYHWD significantly downregulated miR-665 expression(P<0.01),increased the number of autophagosomes.Different doses of BYHWD significantly upregulated protein expression of Bec-lin1(P<0.05,P<0.01),medium and high doses of BYHWD significantly upregulated protein expression of LC3Ⅱ/Ⅰ(P<0.01),and downregulated protein expression of p62(P<0.01).High dose BYHWD significantly upregulated protein expression of DRAM1 in vascular tissues of aging rats(P<0.05).Bioinformatics analysis revealed the presence of specific complementary binding sites between the sequences of miR-665 and DRAM1.Dual-luciferase reporter assays confirmed that miR-665 targeted DRAM1 gene and negatively regulated DRAM1 protein expression.CONCLUSION BYHWD may promote the protein expression of DRAM1 by inhibiting the ex-pression of miR-665,thereby promoting vascular autophagy and delaying vascular aging.

ObjectiveTo observe the effect of Shenling Baizhusan on the intervention of the nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway by regulating ferroptosis in rats with alcoholic liver injury. MethodForty SD rats were randomly divided into model group， polyene phosphatidylcholine group， and high， medium， and low-dose Shenling Baizhusan groups， with 8 rats in each group. Another 8 SD rats were taken as blank group. The model group， polyene phosphatidylcholine group， high， medium， and low-dose Shenling Baizhusan groups were given 10 mL·kg^-1 liquor by gavage for modeling， and the blank group was given equal volume of distilled water by gavage. After 4 h of daily alcoholic administration， 143.64 mg·kg^-1 of polyene phosphatidylcholine group was given to the polyene phosphatidylcholine group， 15， 7.5， 3.75 mg·kg^-1 of Shenling Baizhusan were given to Shenling Baizhusan high， medium， and low-dose groups， respectively， and the blank group and the model group were given equal volume of distilled water. The gavage lasted for 6 weeks. The levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， glutamyl transpeptidase （GGT）， total cholesterol （TC）， and triglyceride （TG） were detected by automatic biochemical analyzer. The levels of tumor necrosis factor-α （TNF-α） and interleukin-β （IL-β） were detected by the enzyme-linked immunosorbent assay （ELISA）. The levels of lipopolysaccharide （LPS）， malondialdehyde （MDA）， superoxide dismutase （SOD）， glutathione （GSH）， and Fe⁺ were detected by biochemical assay. The pathological changes in the liver were observed by hematoxylin-eosin （HE） staining and oil red O staining. The mRNA expression levels of Nrf2， heme oxygenase-1 （HO-1）， glutathione peroxidase 4 （GPX4）， ferritin heavy polypeptide 1 （FTH1）， and nuclear factor-κB （NF-κB） were detected by Real-time polymerase chain reaction （Real-time PCR）. The protein expression levels of Nrf2， HO-1， GPX4， FTH1， p65， and phosphorylation （p）-p65 were detected by Western blot. ResultAs compared with the blank group， the levels of liver function （ALT， AST， and GGT） and blood lipids （TC and TG） in the model group were significantly increased （P<0.05）. The liver showed obvious steatosis， with a large number of fat deposition， the oxidative stress and inflammatory factors were significantly increased （P<0.05）， and the level of Fe⁺ was significantly increased in model group （P<0.05）. The protein expression levels of Nrf2， HO-1， GPX4， and FTH1 was significantly down-regulated （P<0.05）， and those of p65 and p-p65 was significantly up-regulated in the model group （P<0.05）. The mRNA expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly down-regulated （P<0.05）， and the mRNA expression level of NF-κB was significantly up-regulated （P<0.05）. As compared with the model group， the levels of liver function （ALT， AST， and GGT） and blood lipids （TC and TG） in the high-dose and medium-dose Shenling Baizhusan groups were significantly decreased （P<0.05）， liver steatosis was significantly improved， fat deposition was significantly reduced， oxidative stress and inflammatory factors were significantly decreased （P<0.05 ）， and Fe⁺ level was significantly decreased （P<0.05）. In the high-dose and medium-dose Shenling Baizhusan， the protein expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly up-regulated （P<0.05）， and those of p65， p-p65 were significantly down-regulated （P<0.05）. The mRNA expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly up-regulated （P<0.05）， and the mRNA expression level of NF-κB was significantly down-regulated （P<0.05）. ConclusionShenling Baizhusan can effectively reduce liver injury in rats with ALD， regulate steatosis and fat deposition， and play an antioxidant and anti-inflammatory role in the liver. Its mechanism may be related to the inhibition of ferroptosis in hepatocytes by up-regulating the Nrf2 signaling pathway to improve oxidative stress

Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.

Objective:To construct a psychological support training program for palliative care specialist nurses based on positive psychology theory and make a preliminary application.Methods:From August to September, 2020, 142 palliative nursing specialist nurses in Hunan Provincial Palliative Nursing Specialist Training Base were selected as the research objects. Based on positive psychology therapy, using literature review and analysis, semi-structured interviews, expert group meetings and other methods to construct the program, and the work stress, psychological resilience and subjective well-being scores of nurses before and after 8 weeks of psychological support training were compared.Results:The program included three dimensions: cognition, emotion and behavior. After the psychological support training for 8 weeks, the nursing staff′s work stress score 80.76 ± 20.43 was lower than that before the psychological support training 84.70 ± 19.88, the difference was statistically significant( t=6.59, P<0.05), and the psychological resilience score 64.40 ±14.26 was higher than that before the psychological support training 60.19 ±15.85, the difference was statistically significant ( t=-7.39, P<0.05), and the subjective well-being score 75.70 ± 7.70 was higher than that before the psychological support training 74.13 ± 5.98, the difference was statistically significant ( t=-3.52, P<0.05). Conclusions:The psychological support training program based on positive psychology theory can reduce the the work stress, improve psychological resilience and increase subjective well-being of palliative care specialist nurses.

Objective:To explore and optimize the teaching content of clinical case discussion course based on clinical pathophysiology and therapy (CPPT), and to improve the teaching quality of this course.Methods:Self-compiled evaluation form was sent to all the students who participated in this course in 2016-2017 school year, and the teaching satisfaction and teaching characteristics were evaluated. The differences of evaluation results from students with different educational programs or teachers with different seniority were compared, and the correlation between evaluation results and teachers' seniority was analyzed.Results:The students had a high evaluation on the course, but students from different programs had significant different comments on thinking inspiring, speech encouraging, questions guiding, process holding and effective feedback, as well as the self-evaluation of mastery degree. For teachers with different seniority, there were significant differences in scores in all aspects except speech encouraging, but no differences in mastery degree. Spearman analysis showed that there was a certain correlation between scores and teachers' seniority.Conclusion:The overall situation of the CPPT case discussion course in our school is worthy of recognition, but it needs to be improved in many aspects.

Objective: To understand the serum levels of thyroid stimulating hormone (TSH), free triiodothyronine (FT3), free thyroxine (FT4), triiodothyronine (T3), and thyroxine (T4) and identify the related influencing factors of thyroid dysfunction in drug users. Methods: From June to August 2018, a face-to-face questionnaire survey was conducted in 788 male drug users in a drug rehabilitation center in Jiangsu province to collect their socio-demographic information. Then, venous blood sample was collected from each participant for the detection of various hematological indicators, such as thyroid hormones. Results: The abnormal rates of T3, T4, FT3, FT4 and TSH were 4.57%, 1.27%, 0.51%, 0.38% and 0.89%, respectively, in the male drug users. HCV infection was an influencing factor for abnormal T3 level in the male drug users (OR=8.52, 95%CI: 2.36-30.74, P=0.001). And serum T3 (P<0.001) and T4 (P=0.048) levels increased with increasing HCV viral load. Conclusions HCV infection was an influencing factor for the abnormality of serum T3 level in drug users. Therefore, thyroid-related knowledge should be added in the health education for drug users, and the monitoring of thyroid function should be strengthened for drug users infected with HCV.

Objective: To explore associations between physical activity, screen time and anxiety, sleep quality among college students in Shanghai, and to provide a reference for relevant prevention and control. Methods: By using cluster random sampling method, a total of 4 964 students from grade 1 to grade 2 in 3 universities from 3 districts of Shanghai were enrolled. Self-Rating Anxiety Scale, Pittsburgh Sleep Quality Index and International Physical Activity Questionnaire were used to assess the level of anxiety, sleep quality and physical activity. Results: The reporting rate of anxiety symptoms among students was 9.7%(8.7% for males and 11.4% for females) and the prevalence of poor sleep quality was 55.0%(51.8% for males and 60.4% for females), there was significant gender differences in anxiety symptoms and poor sleep quality rate(χ2=9.92, 34.81, P<0.01). Among male students, with adjustment of age, BMI and lifestyle, those who met neither physical activity nor screen time recommendations had 2.23(95%CI=1.31-3.79) and 1.48(95%CI=1.13-1.94) times risks for anxiety and poor sleep quality than those meeting both recommendations. Among girls, there was a significant association between screen time and anxiety(aOR=1.61, 95%CI=1.18-2.21). However, physical activity was not associated with anxiety and sleep quality. Conclusion High screen time and physical inactivity may increase the risk of anxiety and poor sleep quality among male college students, and screen time may also increase the risk of anxiety among female college students.

The family doctor system is a collection of the rules and operating patterns formed during the interaction between the top design of "universal health coverage" and the "contractual experiment" at primary level. Its optimization and upgrading call for continuous attention to primary policy response. A questionnaire survey was made to medical workers at primary healthcare institutions in Zhejiang,to learn the comments and willingness of participation of family doctors for the contractual service policy. The survey found the poor policy response roots in such constraints as lack of effective connection between top level and primary level,poor incentive design,and delayed supporting policies. In view of upgrading supplier policy response,the authors recommended feasible strategies to optimize policy response, namely priori decision-making,process control,both hard and soft tactics,and parallel efforts for both internal and external sides.