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 analyze the clinicopathological characteristics and survival prognosis of young female patients with gastric adenocarcinoma. Methods A retrospective analysis was conducted on female patients who underwent radical gastrectomy at Zhongshan Hospital, Fudan University between January 2014 and December 2020, with postoperative pathological confirmation of gastric adenocarcinoma. Those aged ≤45 years were defined as the young group (n＝287), and were matched in a 1∶2 ratio by pTNM stage with female patients aged ≥60 years (elderly group, n＝574). Clinicopathological characteristics were compared between the two groups. Survival curves were plotted using the Kaplan-Meier method, and overall survival (OS) rates were assessed by log-rank test. Prognostic factors in the young group were analyzed using Cox regression models. Results Compared to elderly patients, young female gastric cancer patients exhibited a higher prevalence of tumors in the middle third of the stomach and a lower proportion in the upper third of the stomach. Molecular profiling revealed a higher frequency of HER2-low expression and elevated Ki-67 index. Pathologically, these patients were more frequently diagnosed with poorly differentiated or undifferentiated adenocarcinoma and signet ring cell carcinoma, while Lauren classification showed a predominance of the diffuse type with a lower proportion of the intestinal type (P＜0.05). Stratified analysis showed no significant difference in OS rates between the two groups for stage Ⅱ and Ⅲ patients; however, among stage Ⅰ patients, the young group had significantly better OS rates than the elderly group (P＝0.037). Multivariate Cox analysis and log-rank test confirmed that pN₃ stage (HR＝3.576, 95%CI 1.652–7.740), stage Ⅲ (HR＝3.581, 95%CI 1.059–12.106), and diffuse type (HR＝2.711, 95%CI 1.316–5.585) were risk factors for poor prognosis in young female gastric cancer patients. Conclusions Young female patients with gastric adenocarcinoma typically present with clinicopathological features such as the diffuse type, poor differentiation, and high proliferation. Moreover, pN₃ stage, stage Ⅲ cancer, and the diffuse type histology are correlated with a poor prognosis in this demographic.

Objective To explore the clinical efficacy of different ligation methods in laparoscopic radi-cal resection of rectal cancer and sigmoid colon cancer and the influencing factors of anastomotic leakage.Methods A total of 82 patients who underwent laparoscopic radical resection of rectal cancer and sigmoid co-lon cancer in this hospital from December 2022 to December 2023 were selected and divided into the low-liga-tion group(n=42)and the high-ligation group(n=40)according to different surgical methods.The inferior mesenteric artery(IMA)classification,IMA length,operation time,intraoperative blood loss,total number of lymph node dissections,number of lymph node dissections in group 253,postoperative hospital stay,exhaust time,incidence of urinary retention,incidence of anastomotic leakage,and incidence of preventive stoma were compared between the two groups.Univariate and multivariate logistic regression analyses were conducted to analyze the influencing factors of anastomotic leakage in patients undergoing radical resection of rectal cancer and sigmoid colon cancer.Results There was no statistically significant difference in IMA length,IMA classi-fication proportion,operation time,intraoperative blood loss,the total number of lymph node dissections,the number of lymph node dissections in group 253,and the postoperative hospital stay between the two groups(P>0.05).Compared with the high-ligation group,the low-ligation group had a shorter exhaust time,lower incidence of anastomotic leakage and preventive ostomy,the differences were statistically significant(P<0.05).Anastomotic leakage occurred in 7 patients.Univariate analysis showed that whether left colic artery(LCA)was preserved,tumor location,presence of underlying diseases,and preventive stoma were associated with anastomotic leakage in patients undergoing radical resection of rectal cancer and sigmoid colon cancer(P<0.05).Multivariate logistics analysis showed that whether LCA was preserved,tumor location,presence of underlying diseases,and presence of preventive stoma were not influencing factors for anastomotic leakage in patients undergoing radical resection of rectal cancer or sigmoid colon cancer(P>0.05).Conclusion In laparoscopic radical resection of rectal and sigmoid colon cancer,the clinical efficacy of preserving LCA is com-parable to that of not preserving LCA,and it can reduce the risk of anastomotic leakage and the probability of preventive stoma,promoting the early recovery of intestinal function.

Objective To determine whether an intelligerct cloud platform follow-up model demonstrates superiority over conventional methods in enhancing follow-up quality,reducing costs,and improving functional recovery following total knee arthroplasty(TKA).Methods A retrospective cohort study was conducted on 260 patients undergoing TKA at our hospital from October 2022 to September 2023.According to the different postoperative follow-up methods,they were divided int a cloud platform group(n=130)and a traditional group(n=130).The baseline data,pre-and postoperative knee function scores,and follow-up costs were collected and compared between the 2 groups.Results The cloud platform group exhibited significantly higher rate of follow-up satisfaction,reduced resource utilization,shorter per-follow-up duration,fewer accompanying persons per visit,lower total follow-up expenditures,and decreased cumulative follow-up time when compared with the traditional group(P<0.05).At 6 weeks,3,6,and 12 months postoperatively,better range of motion(ROM)was observed in the cloud-based group than the traditional group(P<0.05),but no intergroup differences were seen in other knee function scores.Conclusion Cloud platform follow-up enhances early postoperative ROM(6 weeks to 12 months)and demonstrates marked cost-effectiveness when compared to the conventional mode.It represents a viable alternative for post-TKA rehabilitation surveillance.

Objective To explore the enzymatic reaction kinetics of metoprolol tartrate in an in vitro human liver microsome incubation experiment and to explore the effect of famotidine on the metabolism of metoprolol tartrate.Methods An LC-MS/MS method was developed to accurately measure the concentration of metoprolol tartrate in a human liver microsomal incubation system.A human liver microsome incubation system was established to determine the optimal incubation time and protein concentration;using the substrate depletion method,the enzymatic reaction kinetics parameters of metoprolol tartrate(such as Vmax and Km).Series concentrations of famotidine were co-incubated with metoprolol tartrate,and the concentration of metoprolol tartrate was measured to evaluate the effect of famotidine on its metabolism.Results The optimal incubation time for metoprolol tartrate in human liver microsomes was found to be 60 minutes,with an optimal protein concentration of 1.0 mg/mL.The enzymatic reaction kinetics parameters were Vmax=0.07 μmol/min/mg protein and Km=7.84 μmol/L.Conclusion The results indicated that famotidine did not produce a significant inhibitory effect on the metabolism of metoprolol tartrate in human liver microsome incubation system,suggesting that the combination of these two drugs may be relatively safe.

ObjectiveTo investigate the effect of phytoestrogen biochanin A （BCA） on liver fibrosis induced by CCl₄ in female mice with bilateral oophorectomy （ovariectomized） and its mechanism. MethodsA total of 50 ovariectomized Kunming mice were selected and given intraperitoneal injection of CCl₄ to establish a model of liver fibrosis， and then according to body weight， they were randomly divided into model group， positive control group， and low-， middle-， and high-dose BCA groups， with 10 mice in each group. In addition， 10 female mice in the same litter were given resection of a small amount of adipose tissue near both ovaries to establish the sham-operation group. The mice in the positive control group were given estradiol 2 mg/kg by gavage， and those in the low-， middle-， and high-dose BCA groups were given BCA by gavage at a dose of 25， 50， and 100 mg/kg， respectively， once a day for 7 consecutive weeks； the mice in the sham-operation group and the model group were given an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage. The mice were anesthetized and sacrificed after administration to collect samples. Liver index and uterus index were measured； HE staining and Masson staining were used to observe liver histopathological changes； the biochemical analysis was used to measure the activity of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）； ELISA was used to measure the levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in liver tissue， and Western blot was used to measure the relative protein expression levels of collagen Ⅰ， transforming growth factor-beta 1 （TGF-β₁）， alpha-smooth muscle actin （α-SMA）， estrogen receptor beta （ERβ）， and p-NF-κBp65/NF-κBp65 in liver tissue. The t-test was used for comparison of continuous data between two groups； a one-way analysis of various was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and further comparison between two groups. ResultsCompared with the sham-operated group， the model group had a significant increase in liver index and a significant reduction in uterus index， as well as significant increases in the activities of serum AST and ALT， the levels of IL-6 and TNF-α in liver tissue， and the protein expression levels of collagen Ⅰ， TGF-β₁， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in the expression of ERβ in liver tissue （P>0.05）， and the model group showed significant fibrosis lesions in the liver， such as hepatocyte edema， steatosis， and necrosis with inflammatory cell infiltration and hyperplasia， deposition， and staggered distribution of collagen fibers. Compared with the model group， the low-， middle-， and high-dose BCA groups had significant reductions in liver index， the activities of serum AST and ALT， the levels of IL-6 and TNF-α， and the protein expression levels of collagen Ⅰ， TGF-β₁， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in uterine index （P>0.05）， as well as a significant increase in the protein expression level of ERβ in liver tissue （P<0.05） and varying degrees of improvement in liver fibrosis lesions. ConclusionBCA can effectively improve CCL₄-induced liver fibrosis in ovariectomized female mice， possibly by upregulating ERβ to inhibit the NF-κB signaling pathway and then alleviating inflammatory response.

Aim To investigate the impact of Cinobu-facini on the proliferation,invasion,and apoptosis of HepG2 cells and the underlying mechanism.Methods The proliferation of HepG2 cells was assessed using the CCK-8 method following treatment with Cinobufaci-ni.The invasion capability of HepG2 cells was evalua-ted through Transwell assay after exposure to Cinobufa-cini.The apoptosis rates of HepG2 cells post Cinobufa-cini intervention were measured using flow cytometry,and the expression levels of VEGF in the culture medi-um of HepG2 cells were determined using enzyme-linked immunoassay.Furthermore,qRT-PCR and Western blot analyses were conducted to assess the im-pact of Cinobufacini on mRNA and protein expression levels related to the CXCL5/FOXD1/VEGF pathway.The interaction between CXCL5 and FOXD1 was inves-tigated via co-immunoprecipitation.Results Cinobufa-cini treatment led to a gradual decrease in HepG2 cell viability in a dose-dependent manner compared to the control group(P＜0.05).Moreover,Cinobufacini sig-nificantly suppressed HepG2 cell invasion(P＜0.05)while enhancing cell apoptosis(P＜0.05).Notably,Cinobufacini exhibited inhibitory effects on the CX-CL5/FOXD1/VEGF pathway,as evidenced by re-duced expression of related mRNA and proteins(P＜0.05).FOXD1 was identified as the binding site of CXCL5.Overexpression of CXCL5 resulted in in-creased proliferation and VEGF secretion by HepG2 cells(P＜0.05),and increased expression of FOXD1 and VEGF(P＜0.05).However,Cinobufacini inter-vention effectively inhibited liver cancer cell prolifera-tion and invasion(P＜0.05),promoted apoptosis(P＜0.05),reduced VEGF secretion by HepG2 cells(P＜0.05),and downregulated the expression of CXCL5 and FOXD1 in HepG2 cells(P＜0.05);but com-pared with the unexpressed group of Cinobufacini,its ability to inhibit cell activity was weakened(P＜0.05),and its ability to inhibit the expression of CX-CL5,FOXD1,and VEGF was weakened(P＜0.05).Conclusion Cinobufacini may inhibit HepG2 cell pro-liferation and invasion and promote HepG2 cell apopto-sis by regulating the CXCL5/FOXD1/VEGF pathway.

BACKGROUND: Epicardial adipose tissue (EAT) radiomics derived from cardiac computed tomography (CT) images may provide insights into EAT characteristics, which can further predict regression of left ventricular mass index (LVMI) after transcatheter aortic valve replacement (TAVR). This study aimed to develop and validate a radiomics nomogram based on pre-procedural EAT CT to predict inadequate LVMI regression following TAVR. METHODS: Inadequate LVMI regression was defined as ΔLVMI% < 15% at one-year post TAVR. Radiomics features from pre-procedural CT images were selected mainly by least absolute shrinkage and selection operator algorithm. The patients were randomly divided into the training and validation cohorts to establish and evaluate three feature classifier models based on the selected features, using which the Radiomics scores (Radscores) were then calculated. A radiomics nomogram was constructed using independent risk factors and further assessed using area under the curve, calibration curve, and decision curve analysis. RESULTS: A total of 144 consecutive TAVR patients (42 patients with inadequate and 102 patients with adequate LVMI regression) were randomly assigned to the training and validation cohorts (116 patients and 28 patients, respectively). A total of 1130 radiomics features from each patient yielded 6 features for the Radscore construction after selection, with logistic regression and support vector machine models favored. Subsequently, a nomogram based solely on the Radscore was constructed, with an area under the curve of 0.743 in the validation cohort, along with favorable decision curve analysis and calibration curves. CONCLUSIONS The developed radiomics nomogram, serving as a non-invasive tool, achieved satisfactory preoperative prediction of inadequate LVMI regression in TAVR patients, thereby facilitating clinical management.

Humans ; Atrial Fibrillation/surgery* ; Recovery of Function ; Radiofrequency Ablation ; Heart ; Heart Transplantation ; Catheter Ablation ; Treatment Outcome