In current clinical practice, various dermal templates and skin substitutes are used to enhance wound healing. However, the role of wound commensal microbiome in regulating scaffold performance and the healing process remains unclear. In this study, we investigated the influence of both natural and synthetic scaffolds on the wound commensal microbiome and wound repair in three distinct models including diabetic wounds, burn injuries, and germ-free (GF) wounds. Remarkably, synthetic electrospun polycaprolactone (PCL) scaffolds were observed to positively promote microbiome diversity, leading to enhanced diabetic wound healing compared to the natural scaffolds Integra® (INT) and MatriDerm® (MAD). In contrast, both natural and synthetic scaffolds exhibited comparable effects on the diversity of the microbiome and the healing of burn injuries. In GF wounds with no detectable microorganisms, a reversed healing rate was noted showing natural scaffold (MAD) accelerated wound repair compared to the open or the synthetic scaffold (PCL) treatment. Furthermore, the response of the wound commensal microbiome to PCL scaffolds appears pivotal in promoting anti-inflammatory factors during diabetic wound healing. Our results emphasize that the wound commensal microbiome, mediated by different scaffolds plays an important role in the wound healing process.

BACKGROUND: Angiotensin receptor neprilysin inhibitors (ARNIs), angiotensin-converting enzyme inhibitors (ACEIs), angiotensin receptor blockers (ARBs), β-blockers (BBs), and mineralocorticoid receptor antagonists (MRAs) are the cornerstones in treating heart failure with reduced ejection fraction (HFrEF). Sodium-glucose cotransporter 2 inhibitors (SGLT-2is) are included in HFrEF treatment guidelines. However, the effect of SGLT-2i and the five drugs on HFrEF have not yet been systematically evaluated. METHODS: PubMed, Embase, and the Cochrane Library were searched for randomized controlled trials (RCTs) from inception dates to September 23, 2022. Additional trials from previous relevant reviews and references were also included. The primary outcomes were changes in left ventricular ejection fraction (LVEF), left ventricular end-diastolic diameter/dimension (LVEDD), left ventricular end-systolic diameter/dimension (LVESD), left ventricular end-diastolic volume (LVEDV), and left ventricular end-systolic volume (LVESV), left ventricular end-systolic volume index (LVESVI), and left ventricular end-diastolic volume index (LVEDVI). Secondary outcomes were New York Heart Association (NYHA) class, 6-min walking distance (6MWD), B-type natriuretic peptide (BNP) level, and N-terminal pro-BNP (NT-proBNP) level. The effect sizes were presented as the mean difference (MD) with 95% confidence interval (CI). RESULTS: We included 68 RCTs involving 16,425 patients. Compared with placebo, ARNI + BB + MRA + SGLT-2i was the most effective combination to improve LVEF (15.63%, 95% CI: 9.91% to 21.68%). ARNI + BB + MRA + SGLT-2i (5.83%, 95% CI: 0.53% to 11.14%) and ARNI + BB + MRA (3.83%, 95% CI: 0.72% to 6.90%) were superior to the traditional golden triangle ACEI + BB + MRA in improving LVEF. ACEI + BB + MRA + SGLT-2i was better than ACEI + BB + MRA (-8.05 mL/m 2 , 95% CI: -14.88 to -1.23 mL/m 2 ) and ACEI + BB + SGLT-2i (-18.94 mL/m 2 , 95% CI: -36.97 to -0.61 mL/m 2 ) in improving LVEDVI. ACEI + BB + MRA + SGLT-2i (-3254.21 pg/mL, 95% CI: -6242.19 to -560.47 pg/mL) was superior to ARB + BB + MRA in reducing NT-proBNP. CONCLUSIONS: Adding SGLT-2i to ARNI/ACEI + BB + MRA is beneficial for reversing cardiac remodeling. The new quadruple drug "ARNI + BB + MRA + SGLT-2i" is superior to the golden triangle "ACEI + BB + MRA" in improving LVEF. REGISTRATION PROSPERO; No. CRD42022354792.
Humans ; Heart Failure/physiopathology* ; Stroke Volume/physiology* ; Angiotensin Receptor Antagonists/therapeutic use* ; Angiotensin-Converting Enzyme Inhibitors/therapeutic use* ; Sodium-Glucose Transporter 2 Inhibitors/therapeutic use* ; Randomized Controlled Trials as Topic ; Mineralocorticoid Receptor Antagonists/therapeutic use* ; Adrenergic beta-Antagonists/therapeutic use*

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 compare the dose-response effects of single-fraction ultra-high dose rate (FLASH) and conventional dose rate (CONV) whole abdominal irradiation (WAI) with X-rays on acute radiation-induced intestinal injury in mice, in order to identify optimal dose parameters and potential mechanisms.Methods:A total of 186 male C57BL/6J mice were randomly assigned to a non-irradiation group ( n=6), FLASH irradiation groups ( n=90), and CONV irradiation groups ( n=90). Acute radiation-induced intestinal injury models were established using single-fraction WAI with 11, 12, 13, 14, and 15 Gy X-rays (200 Gy/s for FLASH and 4 Gy/min for CONV). Changes in body weight, stool characteristics, and disease activity index (DAI) scores were assessed at 9 d post-irradiation. At 7 d post-irradiation at 11, 12, and 13 Gy, the intestines were collected for macroscopic examination and length measurement. The small intestine was selected for HE staining and quantitative analysis of intestinal crypt number and mucosal epithelial thickness. The survival of mice was assessed at 15 d post-WAI across all dose groups. Results:After single-fraction WAI at 11, 12, and 13 Gy, the body weight was higher in the FLASH group than that in the CONV group ( t=10.17, 12.65, 10.16, P<0.05). The DAI scores for the FLASH group were 1.00±1.10, 3.17±0.75, and 2.83±1.17, respectively, which were lower than those of the CONV group (4.33±0.52, 7.00±0.00, 8.60±0.55; t=8.70, 11.71, 14.99, P<0.05). However, after WAI at 14 Gy and 15 Gy, there were no significant differences in body weight and DAI between the FLASH group and the CONV group ( P>0.05). At 7 d after single-fraction WAI at 11, 12, and 13 Gy, mice in the FLASH group exhibited less intestinal congestion, edema, and shortening compared with the CONV group. The difference between the FLASH and CONV groups were statistically significant in small intestine length at 11 and 13 Gy ( t=4.42, 3.78, P<0.05), and in colorectal length at 11 and 12 Gy ( t=3.97, 3.12, P<0.05). Small intestine HE staining revealed superior preservation of intestinal architecture in the FLASH group compared with the CONV group, characterized by longer villi, increased crypt numbers, thicker mucosal epithelium, and enhanced structural integrity. The differences in crypt number and mucosal epithelial thickness were statistically significant ( tcrypt=13.10, 23.80, 11.90; tmucosal=5.75, 2.64, 7.74; P<0.05). At 15 d post-irradiation, the survival rate in the 15 Gy FLASH group was higher than that in the CONV group (50% vs. 10%, χ2=5.39, P<0.05), with a median survival extension of 6 d ( HR=0.340, 95% CI: 0.115 4-0.999 9). No significant survival differences were observed between the FLASH group and the CONV group at 11, 12, 13, and 14 Gy ( P>0.05). Conclusions:FLASH irradiation significantly alleviated acute radiation-induced intestinal injury from medium single-fraction WAI with 11, 12, and 13 Gy X-rays compared with CONV irradiation, and showed potential to improve mouse survival after single-fraction WAI at 15 Gy. This effect is likely associated with the preservation of intestinal crypts and exhibits a dose-dependent relationship.

Triple-negative breast cancer(TNBC)is a highly aggressive and prognostically unfavorable subtype.Tertiary lymphoid structure(TLS)within the tumor microenvironment,comprising dendritic cells,B cells,T cells,and other immune cells,modulate the tumor immune response.The heterogeneity of TLS in TNBC,such as density,structural maturity,and molecular expression patterns,affects the tumor immune microenvironment and,consequently,treatment responses and clinical outcomes.Studies indicate a positive correlation between the density and maturity of TLS and the pathological complete response(pCR)of TNBC patients,with TLS enhancing the quantity of tumor-infiltrating immune cells and improving anti-tumor immune responses,thereby increasing sensitivity to chemotherapy and immunotherapy.Recent research has found that mature TLS are associated with effective immune responses,becoming significant predictors of treatment response.The combination of TLS with immune checkpoint inhibitors has shown promising prospects.Research demonstrates that promoting the formation or enhancing the functionality of TLS can improve anti-tumor immune effects and enhance treatment outcomes for TNBC patients.Targeting TLS may reduce immune evasion and increase the sensitivity to immunotherapy.However,clinical application of TLS still faces challenges,particularly the impact of their heterogeneity on treatment response.Current assessment methods for TLS are not standardized,lacking a uniform standard and diagnostic system,which limits their widespread application.Future research should focus on resolving these issues by developing standardized assessment tools and further exploring the role of TLS in immune escape and resistance mechanisms.This review aimed to summarize and analyze the existing research progress on TLS in TNBC,in order to provide new ideas for the development of personalized immunotherapy strategies.

The aim of this study is to explore the mechanism of apigenin against transmissible gas-troenteritis virus(TGEV)infection based on network pharmacology and molecular docking.The potential targets of apigenin were obtained from Pharmmapper,Pubchem and other databases.The PubMed database was searched to obtain the relevant targets of TGEV infection.The intersection targets of apigenin and TGEV infection were identified by Draw Venn Diagram online program.A"drug-disease-target"network was constructed using STRING database and Cytoscape 3.8.2 soft-ware.Protein-protein interaction relationships were obtained from the STRING database,and core targets were analyzed.The intersection targets were subjected to GO function and KEGG pathway enrichment analysis using the DAVID database.Finally,the analysis results were validated through molecular docking and in vitro cell experiments.The study identified 431 targets for apigenin,1 177 targets for TGEV infection,and 50 intersection targets for apigenin and TGEV infection.GO enrichment analysis indicated that apigenin was mainly involved in regulating cell differentiation,cell membrane raft formation,apoptosis,and inflammatory responses.The top 15 statistically sig-nificant KEGG enrichment results mainly involve the PI3K-Akt signaling pathway and TNF signa-ling pathway.Docking analysis showed that apigenin had the strongest interaction with matrix metalloproteinase 3(MMP3)with an affinity of-9.5 kJ/mol and the binding activity of MMP3 was the best.The results of in vitro experiments demonstrated that treatment of different concen-trations of apigenin significantly reduced virus titers,virus genome copies,and the expression lev-els of MMP3 and its upstream and downstream proteins compared to the virus-infected group.Api-genin may exert its anti-TGEV effects through multiple targets and pathways,possibly by regula-ting the NF-κB-MMP3-IL-1β signaling pathway.

Triple-negative breast cancer(TNBC)is a highly aggressive and prognostically unfavorable subtype.Tertiary lymphoid structure(TLS)within the tumor microenvironment,comprising dendritic cells,B cells,T cells,and other immune cells,modulate the tumor immune response.The heterogeneity of TLS in TNBC,such as density,structural maturity,and molecular expression patterns,affects the tumor immune microenvironment and,consequently,treatment responses and clinical outcomes.Studies indicate a positive correlation between the density and maturity of TLS and the pathological complete response(pCR)of TNBC patients,with TLS enhancing the quantity of tumor-infiltrating immune cells and improving anti-tumor immune responses,thereby increasing sensitivity to chemotherapy and immunotherapy.Recent research has found that mature TLS are associated with effective immune responses,becoming significant predictors of treatment response.The combination of TLS with immune checkpoint inhibitors has shown promising prospects.Research demonstrates that promoting the formation or enhancing the functionality of TLS can improve anti-tumor immune effects and enhance treatment outcomes for TNBC patients.Targeting TLS may reduce immune evasion and increase the sensitivity to immunotherapy.However,clinical application of TLS still faces challenges,particularly the impact of their heterogeneity on treatment response.Current assessment methods for TLS are not standardized,lacking a uniform standard and diagnostic system,which limits their widespread application.Future research should focus on resolving these issues by developing standardized assessment tools and further exploring the role of TLS in immune escape and resistance mechanisms.This review aimed to summarize and analyze the existing research progress on TLS in TNBC,in order to provide new ideas for the development of personalized immunotherapy strategies.

Objective:To compare the dose-response effects of single-fraction ultra-high dose rate (FLASH) and conventional dose rate (CONV) whole abdominal irradiation (WAI) with X-rays on acute radiation-induced intestinal injury in mice, in order to identify optimal dose parameters and potential mechanisms.Methods:A total of 186 male C57BL/6J mice were randomly assigned to a non-irradiation group ( n=6), FLASH irradiation groups ( n=90), and CONV irradiation groups ( n=90). Acute radiation-induced intestinal injury models were established using single-fraction WAI with 11, 12, 13, 14, and 15 Gy X-rays (200 Gy/s for FLASH and 4 Gy/min for CONV). Changes in body weight, stool characteristics, and disease activity index (DAI) scores were assessed at 9 d post-irradiation. At 7 d post-irradiation at 11, 12, and 13 Gy, the intestines were collected for macroscopic examination and length measurement. The small intestine was selected for HE staining and quantitative analysis of intestinal crypt number and mucosal epithelial thickness. The survival of mice was assessed at 15 d post-WAI across all dose groups. Results:After single-fraction WAI at 11, 12, and 13 Gy, the body weight was higher in the FLASH group than that in the CONV group ( t=10.17, 12.65, 10.16, P<0.05). The DAI scores for the FLASH group were 1.00±1.10, 3.17±0.75, and 2.83±1.17, respectively, which were lower than those of the CONV group (4.33±0.52, 7.00±0.00, 8.60±0.55; t=8.70, 11.71, 14.99, P<0.05). However, after WAI at 14 Gy and 15 Gy, there were no significant differences in body weight and DAI between the FLASH group and the CONV group ( P>0.05). At 7 d after single-fraction WAI at 11, 12, and 13 Gy, mice in the FLASH group exhibited less intestinal congestion, edema, and shortening compared with the CONV group. The difference between the FLASH and CONV groups were statistically significant in small intestine length at 11 and 13 Gy ( t=4.42, 3.78, P<0.05), and in colorectal length at 11 and 12 Gy ( t=3.97, 3.12, P<0.05). Small intestine HE staining revealed superior preservation of intestinal architecture in the FLASH group compared with the CONV group, characterized by longer villi, increased crypt numbers, thicker mucosal epithelium, and enhanced structural integrity. The differences in crypt number and mucosal epithelial thickness were statistically significant ( tcrypt=13.10, 23.80, 11.90; tmucosal=5.75, 2.64, 7.74; P<0.05). At 15 d post-irradiation, the survival rate in the 15 Gy FLASH group was higher than that in the CONV group (50% vs. 10%, χ2=5.39, P<0.05), with a median survival extension of 6 d ( HR=0.340, 95% CI: 0.115 4-0.999 9). No significant survival differences were observed between the FLASH group and the CONV group at 11, 12, 13, and 14 Gy ( P>0.05). Conclusions:FLASH irradiation significantly alleviated acute radiation-induced intestinal injury from medium single-fraction WAI with 11, 12, and 13 Gy X-rays compared with CONV irradiation, and showed potential to improve mouse survival after single-fraction WAI at 15 Gy. This effect is likely associated with the preservation of intestinal crypts and exhibits a dose-dependent relationship.

The aim of this study is to explore the mechanism of apigenin against transmissible gas-troenteritis virus(TGEV)infection based on network pharmacology and molecular docking.The potential targets of apigenin were obtained from Pharmmapper,Pubchem and other databases.The PubMed database was searched to obtain the relevant targets of TGEV infection.The intersection targets of apigenin and TGEV infection were identified by Draw Venn Diagram online program.A"drug-disease-target"network was constructed using STRING database and Cytoscape 3.8.2 soft-ware.Protein-protein interaction relationships were obtained from the STRING database,and core targets were analyzed.The intersection targets were subjected to GO function and KEGG pathway enrichment analysis using the DAVID database.Finally,the analysis results were validated through molecular docking and in vitro cell experiments.The study identified 431 targets for apigenin,1 177 targets for TGEV infection,and 50 intersection targets for apigenin and TGEV infection.GO enrichment analysis indicated that apigenin was mainly involved in regulating cell differentiation,cell membrane raft formation,apoptosis,and inflammatory responses.The top 15 statistically sig-nificant KEGG enrichment results mainly involve the PI3K-Akt signaling pathway and TNF signa-ling pathway.Docking analysis showed that apigenin had the strongest interaction with matrix metalloproteinase 3(MMP3)with an affinity of-9.5 kJ/mol and the binding activity of MMP3 was the best.The results of in vitro experiments demonstrated that treatment of different concen-trations of apigenin significantly reduced virus titers,virus genome copies,and the expression lev-els of MMP3 and its upstream and downstream proteins compared to the virus-infected group.Api-genin may exert its anti-TGEV effects through multiple targets and pathways,possibly by regula-ting the NF-κB-MMP3-IL-1β signaling pathway.

Objective·To explore the correlation between the quantitative results of autofluorescence imaging under computer assistance and the grade of epithelial dysplasia in oral leukoplakia.Methods·From April 2016 to January 2024,357 patients with oral leukoplakia who visited the Department of Oral Mucosal Diseases at Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,were included.Autofluorescence images of the lesions were obtained using a handheld autofluorescence device.These images were converted to grayscale images to obtain quantitative metrics.An ordered multinomial Logistic regression model was fitted in Python,and cumulative probability plots were generated.The dataset was divided into training and testing sets,and a decision tree was generated.Different hyperparameters were adjusted to achieve optimal model performance.Accuracy,precision,and F1 scores were calculated.The model performance was visualized using a confusion matrix.Results·As the degree of epithelial dysplasia increased,the relative mean color level showed a declining trend.In the binary classification of epithelial dysplasia,there was no overlap between the cumulative probability curves of different categories.In the four-category classification,only severe epithelial dysplasia overlapped with other category curves,indicating good discriminative ability of the model.In binary pathological grading,when the training and testing set ratio was 4∶1 and the maximum depth was 2,the accuracy,precision,and F1 scores were 0.792,0.801,and 0.795,respectively.In the four-category pathological grading,when the training and testing set ratio was 9∶1 and the maximum depth was 4,the accuracy,precision,and F1 scores were 0.611,0.537,and 0.569,respectively.Conclusion·Computer-assisted quantitative analysis of autofluorescence images can be used by oral mucosal specialists as a reference to predict the degree of epithelial dysplasia in patients with oral leukoplakia and to monitor their risk of cancer.