BACKGROUND:Nanocell vesicles possess functions such as re-epithelialization,antioxidation,anti-inflammation,and regulation of extracellular matrix remodeling.Meanwhile,apoptotic bodies have the immunomodulatory effects.Therefore,the combination of the two to form nanofusion vesicles can synergistically promote the healing of diabetic skin wounds.OBJECTIVE:To elucidate the impact of nanofusion vesicles on skin wound healing in a diabetic murine model.METHODS:(1)Material preparation and characterization:The primary bone marrow mesenchymal stem cells of C57BL/6J neonatal mice and the neutrophil apoptotic bodies of C57BL/6J mice were isolated and extracted.The nanofusion vesicles were prepared by micro-extrusion mechanism.(2)In vitro experiment:MTT assay was used to detect the proliferative effect of different concentrations of nanofusion vesicles on NIH-3T3 cells and human umbilical vein endothelial cells.Reactive oxygen species fluorescence probe was used to detect the antioxidant effect of nano-fusion vesicles on NIH-3T3 cells treated with hydrogen peroxide(H2O2).The inhibitory effect of nanofusion vesicles on RAW 264.7 macrophage inflammation induced by lipopolyside was detected by real-time quantitative RT-qPCR.(3)In vivo experiment:36 male C57BL/6J mice were employed to develop a murine model of diabetes mellitus.Following the successful induction of diabetes,two circular full-thickness wounds,each with a diameter of 6 mm,were created on either side of the diabetic mice's spine using a skin punch.The mice were divided into three groups by random number table method.The control group was injected with 0.1 mL of phosphate buffer solution.The nanovesicle group was injected with 0.1 mL nanovesicles(25 μg/mL).The nanofusion vesicle group was injected with 0.1 mL nanofusion(25 μg/mL)vesicles.After treatment for three consecutive days,the wound healing and histomorphological changes were observed.RESULTS AND CONCLUSION:(1)In vitro experiment:nanofusion vesicles,when administered at concentrations ranging from 0 to 100 μg/mL,exhibited no toxic effects and promoted the proliferation of NIH-3T3 and HUVEC cell lines.Notably,a concentration of 25 μg/mL nanofusion vesicle significantly enhanced the proliferation of NIH-3T3 cells.Furthermore,the survival rate of human umbilical vein endothelial cells was observed to increase in correlation with escalating concentrations of nanofusion vesicles.Nanofusion vesicles had a good antioxidant effect.In comparison to the H2O2 group,the fluorescence signal indicative of reactive oxygen species was progressively diminished in both the nanovesicle group and the nanofusion vesicle group.Furthermore,nanofusion vesicles possessed anti-inflammatory capabilities,effectively mitigating the inflammatory response in macrophages triggered by lipopolysaccharide stimulation.(2)In vivo experiment:Hematoxylin-eosin and Masson's trichrome staining revealed that in comparison to the control group,both the nanovesicle group and the nanofusion vesicle group exhibited a significant increase in granulation tissue formation and collagen fiber deposition within the wounds by day 6.Notably,the nanofusion vesicle group displayed the most pronounced effects.On day 12,the wound of nanofusion vesicle group was significantly reduced,and the healing rate was significantly faster than that of other groups(P＜0.01),and the effect of promoting wound healing was the most significant.Our findings demonstrated that nanofusion vesicles exhibited superior pro-cell proliferative,antioxidant,and anti-inflammatory properties,thereby exerting a beneficial effect on the promotion of skin wound healing in diabetic mouse models.

Background Public places are important areas for daily human activities. Frequent contact with public items promotes their role as vehicles for microbial spread, creating a substantial risk for the transmission of pathogenic microorganisms. Objective To understand the hygiene status and influencing factors of public items in typical public places in Shanghai from 2010 to 2024, and to provide a scientific basis for optimizing the hygiene management of public items. Methods Based on the monitoring data of public items in public places in Shanghai from 2010 to 2024, the hygiene status was evaluated in three stages: 2010–2019, 2020–2022, and 2023–2024. Chi-square test and logistic regression were used to analyze the impact of factors such as monitoring stages, public place types, and public item categories on the hygiene status. Results The public items in 13807 public places were monitored. A total of 1 022 bed sheets and 9 271 towels were tested for pH value, with qualified rates of 77.66% and 87.81%, respectively. The OR (95%CI) for the pH compliance of towels relative to bed sheets was 1.77 (1.45, 2.15). Compared with bathing places, the ORs for pH compliance of towels in hair salons and beauty salons were 2.24 (1.75, 2.87) and 2.15 (1.57, 2.95), respectively. Additionally, a total of 221 650 samples from various public items were tested for microorganisms, yielding 215554 qualified samples, with an average qualified rate of 97.25%. The qualified rates for total bacterial count, total fungal count, coliforms, and Staphylococcus aureus were 92.66%, 94.63%, 98.85%, and 99.84%, respectively. The logistic regression analysis showed that, compared to the 2016–2019 baseline, no statistically significant differences were observed in the qualified rates of total fungal count across different stages; however, significantly higher qualified rates for total bacterial count, total fungal count, coliforms, and Staphylococcus aureus were recorded during 2020–2022, with ORs (95%CI) of 1.27 (1.16, 1.38), 23.20 (8.42, 63.94), and 12.08 (1.63, 89.92), respectively. Conclusion The pH compliance ranks among the top tier nationwide for cotton textile goods in Shanghai, suggesting minimal detergent retention. Furthermore, continuous improvement in cleaning and disinfection measurements has markedly enhanced the microbial hygiene of public items in public places.

ObjectiveTo explore the effect of oral sodium butyrate on skeletal muscle atrophy in CT26 tumor mice through the gut microbiota-skeletal muscle axis and its potential mechanism. MethodsSixty SPF BALB/c male mice aged 8 weeks were randomly divided into a normal control group (NC, n=18) and a ABX-depleted group (ABX, n=42). The ABX mice were pretreated with a quadruple antibiotic cocktail via oral gavage (0.2 ml per administration, once daily, 6 d per week, for 2 weeks), whereas NC received an equal volume of sterile water. The quadruple antibiotic cocktail consisted of metronidazole (1 g/L), vancomycin (0.5 g/L), ampicillin (1 g/L), and gentamicin (1 g/L). Following successful pretreatment, six mice from each group were randomly selected for gut microbiota sequencing analysis and designated as the Abx group and the NC0 group, respectively. Theremaining mice in ABX were subcutaneously inoculated in the dorsum with 0.2 ml of CT26 cell suspension (at a cell density of 1×10⁷/ml). Then these mice were randomly allocated into three subgroups: a control tumor bearing model group (0_NaB, n=12), a tumor-bearing model group receiving low-dose oral sodium butyrate (L_NaB, n=12), a tumor-bearing model group receiving high-dose oral sodium butyrate (H_NaB, n=12). And mice in NC were inoculated at the same site with 0.2 ml of normal saline. The administration dose for L_NaB was 0.3 g/(kg·d), that for H_NaB was 0.5 g/(kg·d), while NC and 0_NaB were given the same volume of normal saline (0.2ml per time, once daily, 6 d per week, for 4 weeks). The general condition of mice was monitored, and forelimb grip strength gastrocnemius muscle mass and its muscle fiber cross-sectional area were measured for each group. The structural changes in gut microbiota were assessed by 16S rRNA sequencing of cecal contents. Pathological alterations in the intestinal wall were examined via HE staining. Serum and gastrocnemius muscle levels of TNF‑α, IL-6, IL-1β, and LPS were quantified using ELISA. The protein expression of ZO-1 and occludin in the small intestine, as well as proteins associated with the TLR4/MyD88/NF-κB signaling pathway in the gastrocnemius muscle, were detected by Western blot analysis. Results(1) The alpha-diversity in Abx was significantly lower than that in NC0 (P<0.01), a significant decrease of the mass and muscle fiber cross-sectional area of the gastrocnemius (P<0.01), with the majority of gut microbiota being effectively depleted. (2) Compared with NC, the subcutaneous tumors of mice in 0_NaB were prominent, a significant increase of the mass and muscle fiber cross-sectional area of the gastrocnemius, accompanied by a significant decrease in body weight at the end of the 3th and 4th week (P<0.05), and a significant weakening of the forelimb grasping strength at the 5th and 6th week (P<0.01). Compared with 0_NaB, the tumor mass of mice in L_NaB and H_NaB showed a significant decreasing trend, and the grip strength of the forelimbs significantly increased at the 5th and 6th week (P<0.05, P<0.01). (3) Compared with 0_NaB, the Shannon and Observed species indices in α diversity of L_NaB and H_NaB were significantly increased (P<0.05). At the genus level, compared with 0_NaB, L_NaB exhibited a significant decrease in the relative abundance of Parasutterella (P< 0.01), while H_NaB showed significant reductions in the relative abundances of both Escherichia-Shigella and Parasutterella (P < 0.01). (4) Compared with 0_NaB, the small intestinal tissue structure in L_NaB and H_NaB was more intact, the infiltration of inflammatory cells was significantly reduced, and the capillaries were slightly dilated. The expression levels of ZO-1 and occludin proteins in L_NaB were significantly increased (P<0.01). (5) The LPS concentration in the gastrocnemius muscle and the protein expression levels of TLR4, MyD88, p-IκBα, and p-NF‑κB p65 in L_NaB and H_NaB were significantly lower than those in 0_NaB (P<0.05). The serum TNF‑α concentration in H_NaB and TNF-α concentration in the gastrocnemius muscle of the L_NaB and H_NaB were significantly lower than those in 0_NaB (P<0.05, P<0.01, P<0.01). ConclusionOral administration of NaB can improve gut microbiota α diversity, adjusting its composition, improving intestinal mucosal barrier function, reducing the LPS-induced pro-inflammatory response, and delaying skeletal muscle atrophy. The underlying mechanism may involve down regulation of TLR4/MyD88/NF-κB signaling in skeletal muscle.

BACKGROUND:Nanocell vesicles possess functions such as re-epithelialization,antioxidation,anti-inflammation,and regulation of extracellular matrix remodeling.Meanwhile,apoptotic bodies have the immunomodulatory effects.Therefore,the combination of the two to form nanofusion vesicles can synergistically promote the healing of diabetic skin wounds.OBJECTIVE:To elucidate the impact of nanofusion vesicles on skin wound healing in a diabetic murine model.METHODS:(1)Material preparation and characterization:The primary bone marrow mesenchymal stem cells of C57BL/6J neonatal mice and the neutrophil apoptotic bodies of C57BL/6J mice were isolated and extracted.The nanofusion vesicles were prepared by micro-extrusion mechanism.(2)In vitro experiment:MTT assay was used to detect the proliferative effect of different concentrations of nanofusion vesicles on NIH-3T3 cells and human umbilical vein endothelial cells.Reactive oxygen species fluorescence probe was used to detect the antioxidant effect of nano-fusion vesicles on NIH-3T3 cells treated with hydrogen peroxide(H2O2).The inhibitory effect of nanofusion vesicles on RAW 264.7 macrophage inflammation induced by lipopolyside was detected by real-time quantitative RT-qPCR.(3)In vivo experiment:36 male C57BL/6J mice were employed to develop a murine model of diabetes mellitus.Following the successful induction of diabetes,two circular full-thickness wounds,each with a diameter of 6 mm,were created on either side of the diabetic mice's spine using a skin punch.The mice were divided into three groups by random number table method.The control group was injected with 0.1 mL of phosphate buffer solution.The nanovesicle group was injected with 0.1 mL nanovesicles(25 μg/mL).The nanofusion vesicle group was injected with 0.1 mL nanofusion(25 μg/mL)vesicles.After treatment for three consecutive days,the wound healing and histomorphological changes were observed.RESULTS AND CONCLUSION:(1)In vitro experiment:nanofusion vesicles,when administered at concentrations ranging from 0 to 100 μg/mL,exhibited no toxic effects and promoted the proliferation of NIH-3T3 and HUVEC cell lines.Notably,a concentration of 25 μg/mL nanofusion vesicle significantly enhanced the proliferation of NIH-3T3 cells.Furthermore,the survival rate of human umbilical vein endothelial cells was observed to increase in correlation with escalating concentrations of nanofusion vesicles.Nanofusion vesicles had a good antioxidant effect.In comparison to the H2O2 group,the fluorescence signal indicative of reactive oxygen species was progressively diminished in both the nanovesicle group and the nanofusion vesicle group.Furthermore,nanofusion vesicles possessed anti-inflammatory capabilities,effectively mitigating the inflammatory response in macrophages triggered by lipopolysaccharide stimulation.(2)In vivo experiment:Hematoxylin-eosin and Masson's trichrome staining revealed that in comparison to the control group,both the nanovesicle group and the nanofusion vesicle group exhibited a significant increase in granulation tissue formation and collagen fiber deposition within the wounds by day 6.Notably,the nanofusion vesicle group displayed the most pronounced effects.On day 12,the wound of nanofusion vesicle group was significantly reduced,and the healing rate was significantly faster than that of other groups(P＜0.01),and the effect of promoting wound healing was the most significant.Our findings demonstrated that nanofusion vesicles exhibited superior pro-cell proliferative,antioxidant,and anti-inflammatory properties,thereby exerting a beneficial effect on the promotion of skin wound healing in diabetic mouse models.

Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

The prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) is higher among individuals with inflammatory bowel disease (IBD) than in the general population. Emerging evidence indicates that the development of MASLD in patients with IBD may occur independently of traditional metabolic risk factors, suggesting a unique pathophysiological mechanism distinct from conventional MASLD pathways. Bile acids (BAs), which act as critical signaling molecules in enterohepatic circulation, play an essential role in maintaining gastrointestinal homeostasis through bidirectional gut–liver axis communication. These molecules are increasingly recognized as pivotal regulators in the progression of both MASLD and IBD. While previous studies have characterized the dynamics of BA in blood or fecal samples under both conditions, a comprehensive understanding of their metabolic profiles and the associated interactions within the gut–liver axis is still lacking. This review synthesizes current evidence from studies employing BA metabolomics to investigate IBD and MASLD. By focusing on BA signatures, we summarize conserved alterations between these two conditions and their potential mechanisms of disease progression. Our review advances understanding of BA-mediated pathways in IBD and MASLD, providing a foundation for assessing their roles in contributing to the pathogenesis of MASLD in patients with IBD.

In order to explore the possible role and molecular mechanism of the combined action of leech and bear bile in liver and gallbladder diseases, this study first used network pharmacology methods to screen the components and targets of leech and bear bile, as well as the related target genes of liver and gallbladder diseases. The selected key genes were subjected to interaction network and GO/KEGG enrichment analysis. Then, using sodium oleate induced HepG2 cell lipid deposition model and DL-ethionine induced mouse fatty liver model, the activity of leech and bear bile alone and in combination in reducing liver fat was evaluated in vitro and in vivo, and the expression of key pathway related proteins suggested by network pharmacology was detected by Western blot. The results of network pharmacology analysis showed that the active ingredients of leech and bear bile have 295 intersecting targets with liver and gallbladder related diseases, involving more than 200 signaling pathways, including the PI3K/Akt signaling pathway and phospholipase D signaling pathway closely related to glucose and lipid metabolism. The results of in vitro validation experiments showed that both leech and bear bile, alone and in combination, can significantly inhibit the lipid deposition induced by sodium oleate in human liver cells, reduce the triacylglycerol level in cell culture supernatant, and inhibit the lipid content in liver cells. The observation results of Nile red staining confocal microscopy showed that the combination of leech and bear bile had better activity in reducing lipid deposition in liver cells compared to using them alone. In a mouse fatty liver model, the combination of leech and bear bile can better reduce elevated organ indices, blood lipids, and liver lipid levels, as well as lower the levels of serum liver injury biomarkers. The animals used in this experiment and related disposal meet the requirements of animal welfare. Before the experiment, it was reviewed and approved by IACUC, Institute of Materia Medica, Chinese Academy of Medical Sciences. The Western blot experiment results showed that the combination of leech and bear bile can significantly upregulate the expression levels of p-PI3K and p-Akt proteins, and increase the p-PI3K/PI3K and p-Akt/Akt ratios, which is consistent with the predicted results of network pharmacology. The combination of leech and bear bile has great potential for treating fatty liver disease, and activating the PI3K/Akt pathway may be one of the important mechanisms for reducing lipid deposition in liver cells.

Objective To explore the impact of the TINCR-MAF:MAFB transcription factor network on the expression of proliferation and differentiation-related genes in keratinocytes,to verify the role of this network in the occurrence and development of psoriasis and its potential mechanisms.Methods Employed RNA interference technology to knock down TINCR gene expression,and the proliferation ability of keratinocytes was assessed using the CCK-8 method.Additionally,qRT-PCR and Western blot analyses were conducted to evaluate the RNA and protein expression levels of TINCR,MAFB,and KLF4 genes.Immunohistochemical methods were used to detect the expression of KLF4 protein in psoriasis tissues.Results After TINCR gene siRNA interference,the proliferation ability of keratinocytes significantly decreased at 24,48,and 72 hours(P<0.001),indicating that the TINCR gene plays a critical role in cell proliferation.The results of qRT-PCR and Western blot analyses showed that the RNA and protein expression levels of TINCR,MAFB,and KLF4 genes were significantly reduced(P<0.001),suggesting that TINCR may influence the differentiation of keratinocytes by regulating the expression of MAFB transcription factor and KLF4 differentiation-related genes.Furthermore,immunohistochemical results indicated that the expression of KLF4 protein was significantly elevated in psoriasis tissues compared to normal skin tissues,suggesting that KLF4 plays an important role in the pathogenesis of psoriasis.Conclusions The TINCR-MAF:MAFB transcription factor network may participate in the occurrence and development of psoriasis by affecting the proliferation and differentiation of keratinocytes.This finding provides a new perspective on the pathogenesis of psoriasis and potential targets for future therapeutic strategies.

Objective:To investigate the effect of monocular form deprivation (MD) on the distribution density of neuregulin-1 (NRG1) + and NRG1 - parvalbumin (PV) neurons in the primary visual cortex of mice during the critical period of visual development. Methods:Twelve healthy 28-day-old SPF male C57BL/6J mice were randomly divided into control group and MD group by the random number table method, with 6 mice in each group.After MD on postnatal day (P) 28, the MD group was fed until P32, while the control group was fed normally until P32.All mice were sacrificed by cervical dislocation after cardiac perfusion, and brain tissues were quickly collected.After fixation overnight, brain slices were subjected to PV and NRG1 immunofluorescence staining to observe and compare the differences in the distribution density of PV + , PV + /NRG1 + and PV + /NRG1 - neurons in both sides of the primary visual cortex (V1) area of the two groups.This study was conducted in accordance with the Regulations on the Administration of Experimental Animals (2017 Revision), and the study protocol was approved by the Institutional Animal Care and Use Committee of Tianjin Medical University (No.TMUaMEC2022004). Results:Immunofluorescence staining showed that the density of PV + and PV + /NRG1 + neurons was (137.8±4.3), (108.8±4.1), (137.4±4.0)/mm 2 in the contralateral V1 area of the control group, MD group and the ipsilateral V1 area of the MD group, and that of PV + /NRG1 + neurons was (112.0±4.6), (82.1±4.7) and (113.6±5.7)/mm 2, respectively, with statistically significant overall differences ( F=15.88, 12.53; both P<0.001).PV + neuron density and PV + /NRG1 + neuron density in the contralateral V1 area of the MD group were significantly lower than in the control group and in the ipsilateral V1 area of the MD group (all P<0.001).There was no difference in PV + /NRG1 - neuron density between the two groups ( F=0.20, P>0.05). Conclusions:PV + /NRG1 + neurons may be the main cell type regulating the development of primary visual cortex during the critical period.

This study aims to investigate the protective effect of naringenin(Nar)on cadmium(Cd)-induced liver injury in chickens and its regulatory role in endoplasmic reticulum stress(ERS)and autophagy.Forty-eight 1-day-old Hyland Brown egg roosters were divided into four groups:The control group,the Cd group(150 mg/kg CdCl2),the Nar group(250 mg/kg Nar),and the Cd+Nar group(150 mg/kg CdCl2+250 mg/kg Nar),and the rearing period was 60 days.On the 60th day,6 chickens were randomly selected from each group,and after fasting for 24 hours,blood was collected via vein and then removed from the neck and executed,the livers were collected and organ coefficients were calculated,changes in the content of liver function-related indexes were de-termined,pathological sections were made and observed,and the expression of mRNA and protein of related genes were detected by fluorescence quantitative PCR and Western blot method.The re-sults showed that Cd treatment resulted in a highly significant decrease(P＜0.01)in body mass,a highly significant increase(P＜0.01)in liver coefficient,a highly significant increase(P＜0.01)in serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and lactate dehydrogenase(LDH)levels were highly significantly increased(P＜0.01),hepatic antioxidant capacity was highly significantly reduced(P＜0.01),and ERS-related genes(GRP-78,PERK,eIF-2α,CHOP,ATF-4,ATF-6)and autophagy-related genes(Atg-5,Beclin-1,LC3)mRNA expression was highly significantly up-regulated(P＜0.01),and the expression of mRNA for p62 was highly significantly reduced(P＜0.01).After the addition of Nar,Cd-induced liver injury was significantly alleviated,liver function indexes improved,hepatic antioxidant capacity was highly significantly el-evated(P＜0.01),mRNA expression of ERS-related genes(GRP-78,PERK,eIF-2α,CHOP,ATF-4,ATF-6)and autophagy-related genes(Atg-5,Beclin-1,LC3)was highly significantly decreased(P＜0.01)and mRNA expression of P62 was highly significantly increased(P＜0.01).This study suggests that Nar has potential in alleviating Cd-induced liver damage in chickens through the reg-ulation of ERS and autophagy,providing theoretical support for its application in livestock health.