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

Objective To investigate the relationship between serum trimethylamine oxide(TMAO),endo-thelial cell specific molecule 1(Endocan)and cardiac function and pregnancy outcome in patients with hyper-tensive disorders of pregnancy(HDP).Methods A total of 182 patients with HDP admitted to Handan Ma-ternal and Child Health Hospital from January 2021 to June 2023(HDP group)and 98 healthy pregnant women admitted to this hospital during the same period(control group)were selected as research subjects.Serum TMAO,Endocan and left ventricular cardiac function indexes[left ventricular ejection fraction(LVEF),left ventricular end-diastolic volume(LVEDV)and left ventricular end-systolic volume(LVESV)]were compared between the two groups.According to pregnancy outcome,HDP patients were divided into poor outcome group(78 cases)and good outcome group(104 cases).Spearman correlation analysis was used to analyze the correlation between serum TMAO and Endocan and cardiac function indexes in HDP patients,and multi-factor Logistic regression was used to analyze the influencing factors of adverse pregnancy outcomes in HDP patients.The predictive value of serum TMAO and Endocan for adverse pregnancy outcomes in HDP patients was analyzed by receiver operating characteristic(ROC)curve.Results Compared with control group,serum TMAO,Endocan,LVEDV and LVESV were increased in HDP group,and LVEF was decreased(P＜0.05).Serum TMAO and Endocan in HDP patients were negatively correlated with LVEF(P＜0.05),and positively correlated with LVEDV and LVESV(P＜0.05).The incidence of adverse pregnancy outcomes in 182 HDP patients was 42.86％(78/182).Preeclampsia(PE),severe preeclampsia(SPE),24 h urine protein increase,LVEDV increase,LVESV increase,TMAO increase,Endocan increase were independent risk factors for adverse pregnancy outcomes in HDP patients,and LVEF increase was protective factor(P＜0.05).The area under the curve of serum TMAO combined with Endocan in predicting adverse pregnancy outcomes in HDP patients was 0.880,which was greater than 0.793 and 0.788 predicted by serum TMAO and Endocan a-lone.Conclusion The increase of serum TMAO and Endocan levels in HDP patients are relate to the decrease of cardiac function and adverse pregnancy outcomes,and the combined detection of the two has high predictive value for adverse pregnancy outcomes in HDP patients.

Objective To explore the relationship between serum pregnancy-specific protein 1(PSG1),stress-induced protein 2(Sestrin 2),growth arrest-specific protein 6(Gas6)and uterine artery blood flow pa-rameters and fetal growth restriction(FGR)in patients with gestational hypertension(GH).Methods A to-tal of 485 GH patients admitted to Handan Maternal and Child Health Hospital from January 2020 to October 2023 were selected as the research objects and divided into the occurrence group(81 cases)and the non-occur-rence group according to whether FGR occurred.The correlations between serum PSG1,Sestrin 2,Gas6 and uterine artery blood flow parameters[pulse index(PI),resistance index(RI),ratio of peak systolic velocity to end diastolic velocity(S/D)]was analysed,as well as the related influencing factors of FGR in GH patients.In addition,a Nomogram model based on the influencing factors was constructed to analyze the predictive value.Results The serum PSG1 level in occurrence group was significantly lower than that in non-occurrence group,and the serum Sestrin 2,Gas6 levels and PI,RI,S/D values were significantly higher than those in non-occurrence group(P＜0.05).Pearson correlation results showed that serum PSG1 was negatively correlated with the uterine artery blood flow parameters PI,RI,and S/D,and the levels of serum Sestrin 2 and Gas6 were positively correlated with the uterine artery blood flow parameters PI,RI,and S/D(P＜0.05).Gestational di-abetes mellitus,umbilical cord abnormalities,high Sestrin 2,high Gas6,high PI,high RI,and high S/D were independent risk factors for the occurrence of FGR in GH patients(P＜0.05),and increased PSG1 level was protective factor for the occurrence of FGR in GH patients(P＜0.05).Receiver operating characteristic(ROC)curve analysis showed that the area under the curve(AUC)of the Nomogram prediction model con-structed based on the influencing factors for predicting the occurrence of FGR in GH patients was 0.982,and the sensitivity and specificity were 0.943 and 0.938,respectively.The internal verification of the Bootstrap method shows that the Bias-corrected prediction curve basically coincides with the Ideal line,and the consis-tency index(C-index)was 0.964,indicating that the model was relatively stable.The decision curve shows that the threshold probability of this model was 0.01-1.00 and the net return rate was above 0.Conclusion Ser-um PSG1,Sestrin 2 and Gas6 in GH patients are closely related to uterine artery blood flow parameters and FGR,and the three are the influencing factors for the occurrence of FGR in GH patients.The constructed No-mogram model has a good predictive efficacy for FGR.

Objective:To investigate the effects and mechanism of Jianpi Yangzheng Xiaozheng Prescription in lung pre-metastatic niche formation of gastric cancer by regulating the content of programmed death receptor ligand 1 (PD-L1) in gastric cancer exosomes.Methods:Totally 30 615 mice were divided into normal group, model group, Jianpi Yangzheng Xiaozheng Prescription low- and high-dosage groups, and exosome inhibitor group according to random number table method, with 6 mice in each group. The mouse model of lung pre-metastatic niche formation of gastric cancer was established by tail vein injection of MFC cells. After 12 days of administration, the lung metastasis under the intervention of Jianpi Yangzheng Xiaozheng Prescription was evaluated by observing the infiltration of tumor into lung tissue, weighing lung weight and hematoxylin-eosin (HE) staining of lung tissue. Mouse serum exosomes were extracted by ExoQuick kit and identified by transmission electron microscopy and nanoparticle tracking analysis (NTA). The content of PD-L1 in serum exosomes and the expression levels of serum transforming growth factor-β (TGF-β), interleukin-10 (IL-10) and interleukin-6 (IL-6) were detected by enzyme-linked immunosorbent assay (ELISA). Immunofluorescence was used to detect the phenotype of myeloid-derived suppressor cells (MDSC) and tumor-associated macrophages (TAM) and the expression of PD-L1 in MDSC and TAM.Results:Compared with the model group, after the intervention of Jianpi Yangzheng Xiaozheng Prescription, the lung metastases of mice were reduced ( P<0.05), and the weight of metastatic tumors decreased ( P<0.05). PD-L1 in serum exosomes and the proportion of MDSC and M2 TAM in lung tissue microenvironment decreased, as well as the expression of PD-L1 on MDSC and TAM decreased ( P<0.05). The serum levels of IL-10 and IL-6 significantly decreased ( P<0.05). Conclusion:Jianpi Yangzheng Xiaozheng Prescription can reduce the proportion of MDSC and M2 TAM and the expression level of PD-L1 in the microenvironment of lung tissue before metastasis by inhibiting the transmission of gastric cancer exosome PD-L1 to MDSC and TAM, and reduce the contents of inflammatory factorsIL-10 and IL-6, so as to play a role in improving the microenvironment before lung metastasis of gastric cancer.

ObjectiveTo analyze the characteristics and changing trend of injury cause of mortality of residents in Qingpu District from 2002 to 2020， and to provide scientific reference for formulating regional prevention and control measures. MethodsThe injury mortality data of the registered residents in Qingpu District from 2002 to 2020 were collected. The indicators such as crude mortality rate， standardized mortality rate， and the ranking of causes of death were calculated. ResultsFrom 2002 to 2020， the average annual crude mortality rate was 50.27/100 000， the age-standardized mortality rate based on the world standard population（ASRW） was 30.08/100 000， and the age-standardized mortality rate based on the 2010 Chinese census（ASMRC） was 35.58/100 000. The average annual crude mortality rate of males was higher than that of females ［Z=54.402， Mantel-Hanszel χ²=1 742.509， P<0.01）. The overall injury mortality rate showed a downward trend with an average annual percent change（AAPC）of -4.07% （95%CI： -5.23%‒-2.90%）， P<0.001］. The top four causes of injury death were transportation accident， indeliberate fall， drowning， and suicide. The leading causes of death in 0‒ years old， 15‒ years old and ≥65 years old were drowning， transportation accident and indeliberate fall， respectively. The ASRW of transportation accident， drowning and suicide all showed a decreasing trend， and the AAPC were -8.22% （95%CI： -10.16%‒-6.24%）， -6.99% （95%CI： -9.68%‒-4.22%） and -6.21% （95%CI： -9.38%‒-2.94%）， respectively. ConclusionThe injury death rate of residents in Qingpu District shows a decreasing trend， and the distribution characteristics of injury death are different among different genders and age groups. Corresponding prevention and control strategies should be adopted for different populations.

Objective:To describe the prevalence of food allergy among children aged 0-5 years in China and to explore related influencing factors.Methods:Multistage stratified random sampling method was used to collect data from 275 surveillance sites of the China National Nutrition and Health Survey of Chinese children and lactating mothers programs in 31 provinces (autonomous regions and municipalities) of China in 2016-2017. A total of 70 107 participants aged 0-5 years were included in this study. The study collected information of participants' demographic characteristics and food allergies by face-to-face questionnaire. The prevalence of food allergy was analyzed, using the complex data weighting method. The logistic regression models were used to analyze the influencing factors related to food allergy.Results:The overall prevalence of self-reported food allergy among children aged 0-5 years was 4.81%. Prevalence rates in infants aged 0-5 months, and 6-23 months and preschool children aged 2-5 years were 0.81%, 4.68% and 5.26%, respectively. The results of logistic analysis showed that there was a significantly positive correlation between factors including children from 6 months to 5 years old, urban area, southwest area, first-born, mothers with college education or above, and the prevalence of food allergy in children. Shrimp, poultry eggs, crab shellfish, fruit, milk and fish appeared the common allergic foods in children aged 0-5 years, with prevalence rates of self-reported food allergy as 1.55%, 1.25%, 0.99%, 0.97%, 0.87% and 0.86%, respectively. The proportion of single food allergy in children with allergies was 69.85%.Conclusions:Among children aged 0-5 years, the prevalence of self-reported food allergy increases with age, in China. Foods that is prone to allergies include fish, shrimp, crab, shellfish, poultry eggs, milk and fruits, etc. Most allergies were only caused by single food in children, under observation.

Background::Hypothermia therapy has been suggested to attenuate myocardial necrosis; however, the clinical implementation as a valid therapeutic strategy has failed, and new approaches are needed to translate into clinical applications. This study aimed to assess the feasibility, safety, and efficacy of a novel selective intracoronary hypothermia (SICH) device in mitigating myocardial reperfusion injury.Methods::This study comprised two phases. The first phase of the SICH was performed in a normal porcine model for 30 minutes ( n = 5) to evaluate its feasibility. The second phase was conducted in a porcine myocardial infarction (MI) model of myocardial ischemia/reperfusion which was performed by balloon occlusion of the left anterior descending coronary artery for 60 minutes and maintained for 42 days. Pigs in the hypothermia group ( n = 8) received hypothermia intervention onset reperfusion for 30 minutes and controls ( n = 8) received no intervention. All animals were followed for 42 days. Cardiac magnetic resonance analysis (five and 42 days post-MI) and a series of biomarkers/histological studies were performed. Results::The average time to lower temperatures to a steady state was 4.8 ± 0.8 s. SICH had no impact on blood pressure or heart rate and was safely performed without complications by using a 3.9 F catheter. Interleukin-6 (IL-6), tumor necrosis factor-α, C-reactive protein (CRP), and brain natriuretic peptide (BNP) were lower at 60 min post perfusion in pigs that underwent SICH as compared with the control group. On day 5 post MI/R, edema, intramyocardial hemorrhage, and microvascular obstruction were reduced in the hypothermia group. On day 42 post MI/R, the infarct size, IL-6, CRP, BNP, and matrix metalloproteinase-9 were reduced, and the ejection fraction was improved in pigs that underwent SICH.Conclusions::The SICH device safely and effectively reduced the infarct size and improved heart function in a pig model of MI/R. These beneficial effects indicate the clinical potential of SICH for treatment of myocardial reperfusion injury.

Objective:To study and screen the differential expression of inflammatory proteins in diabetes skin ulcers and common skin ulcers, so as to provide experimental basis for further research on anti-inflammatory and healing drug targets of diabetes skin ulcers.Methods:The tissues of 11 patients with diabetes skin ulcer, 12 patients with common skin ulcer and 11 patients with normal skin were collected from the First Hospital of Hunan University of Chinese Medicine. The levels of inflammatory protein Toll like receptor 4 (TLR4), nuclear factor κB (NF-κB), pro-inflammatory factor interferon -γ (IFN -γ), tumor necrosis factor - α (TNF -α), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-1β (IL-1β), macrophage chemotactic protein-1 (MCP-1), anti-inflammatory factors epidermal growth factor (EGF), interleukin-4 (IL-4), and interleukin-10 (IL-10) were detected in three groups of tissues using enzyme-linked immunosorbent assay (ELISA).Results:Compared with normal tissues, the concentrations of TLR4, NF-κB, IFN -γ, TNF -α, IL-1β, IL-6, IL-8, MCP-1 and EGF in common ulcer skin tissues and diabetes ulcer tissues were higher, and the concentrations of IL-10 were lower, with statistically significant differences (all P<0.05); Compared with the normal tissue, the concentration of IL-4 in diabetes ulcer tissue was lower, the difference was statistically significant ( P<0.05); Compared with ordinary ulcer skin tissue, the concentrations of TLR4, NF-κB and MCP-1 in diabetes ulcer tissue were higher, and the concentrations of IL-4 were lower, with statistically significant differences (all P<0.05). Conclusions:The skin ulcer in diabetes patients will have inflammatory reaction, and high glucose promotes the inflammatory reaction of skin ulcer, which may be related to the abnormal expression of TLR4, NF-κB, MCP-1 and IL-4. TLR4/NF-κB signal pathway and inflammatory factors MCP-1 and IL-4 may be the target of the inflammation regulation of diabetes skin ulcer.

ObjectiveTo investigate the possible mechanism of Ruyi Heibai Power （如意黑白散， RHP） in the treatment of vitiligo. MethodsTwenty-four C57BL/6 mice were randomly divided into blank group， model group， high-dose and low-dose RHP groups， with 6 mice in each group. Model group， high- and low-dose RHP groups were all applied hydroquinone to establish vitiligo animal model. After modeling， High- and low-dose RHP groups were given 7.02 g/kg and 2.34 g/kg of RHP by gavage， respectively， while the blank group and model group were intragastrically given 10ml/kg of normal saline， once a day for 36 days. After administration， the skin lesions were observed with naked eye， and HE staining was used to observe the melanin content of the skin lesions. Immunohistochemistry was used to determine the expression of CD3⁺ and CD8⁺ T cells in skin tissue. Immunofluorescence staining was used to measure the expression of programmed death receptor 1 （PD-1） in the skin lesion tissue. RT-PCR was used to detect programmed cell death ligand 1 （PD-L1） mRNA expression. ELISA was used to detect serum superoxide dismutase （SOD）， tumor necrosis factor （TNF-α）， and tyrosinase （TYR） levels. ResultsCompared to those in the blank group， the skin of the mice in the model group was pale， and the melanin content was significantly reduced under the microscope after HE staining； the rate of excellent and good skin lesions decreased， and the melanin granules in the cells around the epidermis and hair follicles decreased significantly； the expression of CD3⁺ and CD8⁺ T cells in skin tissue increased significantly， and the expressions of PD-L1 mRNA and PD-1 decreased； the content of TYR decreased， while the content of SOD and TNF-α increased （P＜0.05）. Compared to those in the model group， the skin color of high- and low-dose RHP groups were deepened， and the melanin content increased； the rate of excellent and good skin lesions increased， as well as the melanin granules in the spinous cell layer， basal cells and hair follicles； the expression of CD3⁺ and CD8⁺ T cells in the skin lesions decreased， while PD-L1 mRNA and PD-1 expression increased； the content of TYR increased， while the content of SOD and TNF-α decreased （P＜0.05）. Compared to the low-dose RHP group， the high-dose group had a larger pigment recovery area in the modeling area， an increased rate of excellent and good skin lesions， an increase in spinous cell layer， basal cells， and hair follicle melanin granules， a decrease in CD3⁺ and CD8⁺ T cells expression， an increase in the expression of PD-L1 mRNA and PD-1， an elevated TYR content， and decreased SOD and TNF-α contents （P＜0.05）. ConclusionRHP can increase skin melanin content of vitiligo mice.The mechanism of action may be related to activating the PD-1/PD-L1 signaling pathway， and then reducing the destruction of melanocytes by T cell-mediated autoimmunity.

Lipotoxicity is a pivotal factor that initiates and exacerbates liver injury and is involved in the development of metabolic-associated fatty liver disease (MAFLD). However, there are few reported lipotoxicity inhibitors. Here, we identified a natural anti-lipotoxicity candidate, HN-001, from the marine fungus Aspergillus sp. C1. HN-001 dose- and time- dependently reversed palmitic acid (PA)-induced hepatocyte death. This protection was associated with IRE-1α-mediated XBP-1 splicing inhibition, which resulted in suppression of XBP-1s nuclear translocation and transcriptional regulation. Knockdown of XBP-1s attenuated lipotoxicity, but no additional ameliorative effect of HN-001 on lipotoxicity was observed in XBP-1s knockdown hepatocytes. Notably, the ER stress and lipotoxicity amelioration was associated with PLA2. Both HN-001 and the PLA2 inhibitor MAFP inhibited PLA2 activity, reduced lysophosphatidylcholine (LPC) level, subsequently ameliorated lipotoxicity. In contrast, overexpression of PLA2 caused exacerbation of lipotoxicity and weakened the anti-lipotoxic effects of HN-001. Additionally, HN-001 treatment suppressed the downstream pro-apoptotic JNK pathway. In vivo, chronic administration of HN-001 (i.p.) in mice alleviated all manifestations of MAFLD, including hepatic steatosis, liver injury, inflammation, and fibrogenesis. These effects were correlated with PLA2/IRE-1α/XBP-1s axis and JNK signaling suppression. These data indicate that HN-001 has therapeutic potential for MAFLD because it suppresses lipotoxicity, and provide a natural structural basis for developing anti-MAFLD candidates.