Background and aims:Metabolic dysfunction-associated fatty liver disease(MAFLD)is a common chronic condition that can lead to cancer due to its complex pathogenesis.Therapeutic agents targeting AMP-activated protein kinase(AMPK)activation have been suggested as potential treatments for metabolic disorders such as metabolic dysfunction-associated steatohepatitis(MASH).Rhizoma Atractylodis Mac-rocephalae(RAM)has been clinically used to treat obesity-related health problems,but its therapeutic effects on MAFLD and the underlying mechanism remain unclear.Therefore,this study was conducted to evaluate the function and underlying mechanism of RAM in the treatment of MAFLD.Methods:The effect of RAM decoction on MAFLD was evaluated using a high-fat diet(HFD)-induced MAFLD mouse model.In vitro studies were conducted using a palmitic acid/oleic acid-induced lipid accumulation model in the alpha mouse liver 12 cells and RAM-containing serum.The underlying mechanisms were elucidated through a combination of network pharmacology analysis,immunohis-tochemistry,western blotting,and polymerase chain reaction analysis.Results:Administration of RAM decoction significantly reduced body weight gain in MAFLD mice without changing food intake.The weights of the liver and inguinal adipose tissues were also reduced after RAM treatment.Additionally,RAM administration decreased serum levels of alanine aminotrans-ferase,aspartate transaminase,total cholesterol,triglyceride,low-density lipoprotein cholesterol,and glucose,while reducing lipid droplet accumulation in the liver tissues of MAFLD mice.The underlying mechanisms included the activation of the phosphorylation of AMPK and acetyl-CoA carboxylase(ACC),and inhibition of the expression of sterol regulatory element binding protein 1(SREBP1).However,RAM did not alter the protein expression levels of peroxisome proliferator-activated receptor α and carnitine palmitoyltransferase-1α.Furthermore,the RAM-induced upregulation of phosphorylated AMPK,phos-phorylated ACC,and SREBP1 expression,as well as the downregulation of fatty acid synthase expression,were reversed by using an AMPK inhibitor.Conclusions:Through a combination of network pharmacology and experimental validation,we demonstrated that RAM may exert therapeutic effects on MAFLD by inhibiting lipid synthesis and activating phosphorylated AMPK pathways.

Objective To analyze the CT signs of acute blunt or penetrating intestinal injury,and to improve the diagnostic accuracy of multi spiral CT for intestinal injury.Methods The CT and clinical data of 63 patients with intestinal injury confirmed by clinical surgical exploration who underwent emergency CT scan were collected,and the CT findings and surgical findings were comparatively analyzed.Results There were 63 cases of intestinal injury,of which 26 cases were complicated with mesenteric injury.The direct CT signs of intestinal injury included intestinal wall thickening sign and intestinal discontinuity sign,which accounted for 64%(40/63)and 17%(10/63),respectively.The indirect CT signs of intestinal injury included intraperitoneal/retroperitoneal gas sign,intraperitoneal/retroperitoneal effusion sign,intramural air,and portal venous gas,which accounted for 72%(45/63),88%(55/63),7%(5/63)and 5%(3/63),respectively.Conclusion Recognizing the CT signs of intestinal injury,such as intestinal wall thickening sign,intestinal discontinuity sign,intraperitoneal/retroperitoneal gas sign,intraperitoneal/retroperitoneal effusion sign,intramural air,and portal venous gas can help to make the early and correct diagnosis of intestinal injury if combined with clinical practice.

BACKGROUND:The treatment for bacterial biofilms after internal fixation surgery is a very difficult problem in clinic.It is a great significance to establish an animal model of irrigation for treating bacterial biofilms in the early stage after internal fixation surgery. OBJECTIVE:To establish an animal model for treating bacterial biofilms with different drugs through irrigation in early stage after internal fixation surgery. METHODS:Six New Zealand white rabbits were selected.Bilateral femoral surfaces were exposed and drilled holes were made,and bone plates colonized with Pseudomonas aeruginosa(experimental group)and blank bone plates(blank control group)were implanted around the drilled holes on one side,and two drainage tubes were retained and fixed to serve as the"inlet"and"outlet,"respectively.The model was immersed for a certain period of time after simulated perfusion before rinsing.After the simulated irrigation,the plates were soaked for a certain time before washing.At 5 days postoperatively,the rabbits were observed for body temperature,wound condition,bacterial culture of drainage fluid,and crystalline violet staining and scanning electron microscopy of the bone plate. RESULTS AND CONCLUSION:Six rabbits had difficulty in moving the affected limbs after surgery and showed elevated body temperature at 2-4 days after surgery.Local swelling could be touched at some wounds in the experimental group,and the wounds in the blank control group healed well.The results of bacterial culture of drainage fluid showed that Pseudomonas aeruginosa diffused or spread in the experimental group.At 5 days after surgery,the plate in the experimental group became purple shown by crystalline violet staining,and the absorbance value at 570 nm detected by the microplate reader was 2.621±0.088,indicating the presence of bacteria.Scanning electron microscopy at 5 days after surgery showed that a large number of bacterial microcolonies appeared on the surface of the plate in the experimental group,forming a highly inhomogeneous three-dimensional structure similar to the"mushroom-like"and"tower-like"structures,with filamentous water channels connecting the"mushroom-like"structures,which were typical biofilm structures with high densities,while no obvious colonies were seen in the blank control group.Overall,this animal model simulates the state of infected biofilm formation due to early infection after internal fixation and provides an available method of irrigation with different drugs.

Purpose To construct and validate a machine learning-based diagnostic model for distinguishing between Mycobacterium avium-intracellular complex pulmonary disease(MAC-PD)and pulmonary tuberculosis(PTB)via chest CT images.Materials and Methods Retrospective data from patients diagnosed with MAC-PD and PTB between May 2021 and August 2022 at Beijing Chest Hospital,Capital Medical University,which were collected as the training set.The prospective external validation set was obtained from patients at the First Affiliated Hospital of Henan University of Chinese Medicine between September 2022 and May 2023.Clinical and radiological data were analyzed,and multivariable logistic regression,random forest and support vector machine(SVM)models were established and externally validated using the validation set.The diagnostic performance of models were evaluated using receiver operating characteristic curve and precision-recall curve,and the differences of the areas under the curve of various models were compared via the Delong test.Results There were significant differences in age and hemoptysis rate between the two groups(t=30.414,P<0.001;χ2=6.186,P=0.013).There were statistically significant differences in cavity types and morphology between the two groups(χ2=6.546,P=0.011;χ2=24.113,P<0.001),but there was no significant difference in the distribution and characteristics of cavitary lesions(P>0.05).There were significant differences in the types and distribution of bronchiectasis between the two groups(χ2=4.634,P=0.031;χ2=23.145,P<0.001).Compared with logistic regression and random forest models,the SVM model had better differential diagnostic performance,and the area under the receiver operating characteristic curve,sensitivity,specificity,accuracy,positive predictive value and negative predictive value were 0.960(95%CI 0.935-0.985),85.7%,93.6%,90.5%,93.3%,88.0%and 0.885(95%CI 0.803-0.967),respectively,76.7%,80.0%,78.3%,79.3%,77.4%.The precision-recall curve showed that the SVM model had high precision and low recall,that was,the model performs well.Conclusion The machine learning-based models exhibits excellent diagnostic performance and can assist in differentiating MAC-PD and PTB.

Surgical resection and radiotherapy have been commonly applied to the treatment of early- and intermediate-stage Merkel cell carcinoma. In recent years, immunotherapy based on immunogenic characteristics of Merkel cell carcinoma has been proved to be effective in the treatment of advanced Merkel cell carcinoma. This review focuses on advances in the treatment of Merkel cell carcinoma, and provides a reference for its clinical diagnosis and treatment.

This study aims to construct C57/B6-L and A549 cell lines that stably overexpress circu-lar RNA LAMP3(circLAMP3),laying the foundation for further research on the biological func-tions of circLAMP3.Total RNA was extracted and reverse transcripted into cDNA from C57/B6-L and A549 cells to amplify the full-length sequence of circLAMP3.Then,the fragments of cir-cLAMP3 were ligated into pLC5-ciR vector to obtain pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 recombinant plasmids.The lentiviruses expressing circLAMP3 were packaged on tran-sient transfected HEK293T cells.C57/B6-L and A549 cells were infected with lentiviruses to gen-erate cell lines overexpressing circLAMP3 through puromycin screening.To verify the overexpres-sion efficiency of circLAMP3 of cell lines,we performed the fluorescence microscopy,PCR amplifi-cation,quantitative PCR(qPCR),and Sanger sequencing experiments.The results indicated that the overexpression plasmids of pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 were suc-cessfully constructed.Strong green fluorescence was observed under a fluorescence microscopy.C57/B6-L and A549 cell lines showed a significant increase in the expression of circLAMP3 by PCR and qPCR methods.Sanger sequencing results showed that the junction site of circLAMP3 was correct.This study successfully constructed C57/B6-L and A549 cell lines overexpressing circLAMP3,providing biomaterials for further exploration of the biological function of circLAMP3 in influenza virus replication.

Background/Aims: Metabolic dysfunction-associated steatotic liver disease (MASLD) has become an increasingly important health challenge, with a substantial rise linked to changing lifestyles and global obesity. Ursolic acid, a natural pentacyclic triterpenoid, has been explored for its potential therapeutic effects. Given its multifunctional bioactive properties, this research further revealed the pharmacological mechanisms of ursolic acid on MASLD. Methods: Drug target chips and bioinformatics analysis were combined in this study to explore the potential therapeutic effects of ursolic acid on MASLD. Molecular docking simulations, surface plasmon resonance analyses, pull-down experiments, and co-immunoprecipitation assays were used to verify the direct interactions. Gene knockdown mice were generated, and high-fat diets were used to validate drug efficacy. Furthermore, initial CD4+ T cells were isolated and stimulated to demonstrate our findings. Results: In this study, the multifunctional extracellular matrix phosphorylated glycoprotein secreted phosphoprotein 1 (SPP1) was investigated, highlighting its capability to induce Th17 cell differentiation, amplifying inflammatory cascades, and subsequently promoting the evolution of MASLD. In addition, this study revealed that in addition to the canonical TGF-β/IL-6 cytokine pathway, SPP1 can directly interact with ITGB1 and CD44, orchestrating Th17 cell differentiation via their joint downstream ERK signaling pathway. Remarkably, ursolic acid intervention notably suppressed the protein activity of SPP1, suggesting a promising avenue for ameliorating the immunoinflammatory trajectory in MASLD progression. Conclusions Ursolic acid could improve immune inflammation in MASLD by modulating SPP1-mediated Th17 cell differentiation via the ERK signaling pathway, which is orchestrated jointly by ITGB1 and CD44, emerging as a linchpin in this molecular cascade.

Specific tumor-targeted gene delivery remains an unsolved therapeutic issue due to aberrant vascularization in tumor microenvironment (TME). Some bacteria exhibit spontaneous chemotaxis toward the anaerobic and immune-suppressive TME, which makes them ideal natural vehicles for cancer gene therapy. Here, we conjugated ZIF-8 metal-organic frameworks encapsulating eukaryotic murine interleukin 2 (Il2) expression plasmid onto the surface of VNP20009, an attenuated Salmonella typhimurium strain with well-documented anti-cancer activity, and constructed a TME-targeted Il2 delivery system named Il2/ZIF-8@Salmonella. Both in vitro and in vivo experiments demonstrated that Il2/ZIF-8@Salmonella maintained the tumor-targeting feature of bacteria, and could be effectively phagocytosed by intratumoral macrophages, thus leading to the expression and secretion of IL2 in TME. The detailed analysis of tumor immune microenvironment (TIME) showed that one dose of combinatorial Il2/ZIF-8@Salmonella achieved synergistic actions on a potent remodeling of TIME, marked by the activation of cytotoxic T cells and M1-polarization of macrophages in TME, thus leading to significant anti-tumor effects in melanoma, orthotopic hepatocellular carcinoma, and pulmonary metastasis models. More importantly, Il2/ZIF-8@Salmonella exhibited high safety to major organs and hematopoietic systems. Taken together, we report a novel plasmid/ZIF-8@Salmonella system that simultaneously achieves effective TME-targeted delivery of therapeutic gene, as well as synergistic re-activation of TIME.

Background and aims:Kehuang(KH)capsule is an herbal medical product approved for the treatment of liver diseases,including liver injury,in China.However,the mechanism is still unclear.This study aimed to elucidate the protective effects of KH capsule against carbon tetrachloride(CCl4)-induced acute liver injury(ALI)in a murine model.Methods:Mice were randomly divided into control,model(CCl4),CCl4+KH_Low and CCl4+KH_High group.Liver enzyme levels and histological changes were assessed to evaluate liver injury.Oxidative stress markers and inflammatory cell infiltration in liver tissues were measured.Additionally,network pharmacology was employed to explore the potential mechanisms of KH capsule.Results:KH capsule significantly reduced serum alanine aminotransferase(ALT)and aspartate amino-transferase(AST)levels,as well as the necrotic area in liver tissue.KH capsule also decreased the infil-tration of macrophages and neutrophils,thereby inhibiting the expression of interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α),and interleukin-1 beta(IL-1β).Furthermore,KH capsule decreased liver malondialdehyde(MDA)levels and increased superoxide dismutase(SOD)activity.The number of ter-minal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)-positive cells in liver tissue was also reduced.The expression of nuclear factor erythroid 2 related factor 2(Nrf2)and heme oxygenase-1(HO-1)proteins was significantly elevated,while the protein expression of cyto-chrome P450 2E1(CYP2E1)was significantly reduced.Mass spectrometry identified genistein,galangin,wogonin,skullcapflavone Ⅱ,and hispidulin as potential active ingredients of KH capsule.Network pharmacology analysis revealed enrichment in the mitogen-activated protein kinase(MAPK)and phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT)signaling pathways.Western blot analysis confirmed that KH capsule suppressed AKT,extracellular signal-regulated kinase(ERK),and p38 signaling.Conclusions:These findings suggest that KH capsule could exert protective effects against CCl4-induced ALI,with the inhibition of MAPK and PI3K-AKT signaling pathways playing a crucial role in its mecha-nism of action.

Objective To explore the efficacy of the resolving depression and tranquilizing herbal sachets in alleviating sleep disorders of soldiers in naval vessels. Methods Fifty-nine soldiers with sleep disorders (PSQI scale score ≥8 and SAS scale score ≥50) were selected before training at sea, and were randomly divided into sachet treatment group, mindfulness meditation group and control group. During the training period, the sachet treatment group was given traditional Chinese medicine sachet treatment, the mindfulness meditation group was trained in mindfulness meditation, and the control group was not given any intervention. Results Compared with the control group, the PSQI and SAS scores of soldiers in both the experimental sachet treatment group and the positive meditation group were significantly improved; compared with the pre-intervention data, the PSQI and SAS scores of both the positive meditation group and the sachet treatment group were significantly decreased, and the degree of decrease was comparable. Conclusion The homemade improving depression and tranquilizing TCM sachets have the effect of alleviating the sleep disorders of naval troops served on vessels, which is equivalent to mindfulness meditation. It is easy to use and operate, which is suitable for promotion in the military.