ObjectiveTo investigate the potential targets of 2，5-dihydroxybenzoic acid （2，5-DHBA） in the treatment of metabolic dysfunction-associated fatty liver disease （MAFLD） and the molecular mechanism by which it regulates the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. MethodsA network pharmacology analysis was performed at first， and related databases were used to obtain the common action targets of 2，5-DHBA and MAFLD， followed by molecular docking and pathway enrichment analysis to predict the potential biological processes and signaling pathways regulated by these targets. Then mouse normal hepatocytes AML-12 were used for cell experiments， and CCK-8 assay was used to determine the three optimal intervention concentrations of 2，5-DHBA. AML-12 cells were divided into control group， model group （cells cultured in a medium containing 2 mmol/L free fatty acid ［FFA］ to establish a cell model of MAFLD）， and three 2，5-DHBA intervention groups （cells cultured with FFA and 2，5-DHBA at the three different concentrations of 1.25， 5， and 20 μmol/L， respectively）. Oil red O staining was used to observe intracellular lipid accumulation； the fluorescent probe DCFH-DA was used to measure the level of intracellular reactive oxygen species （ROS）； biochemical assays were used to measure the content of triglyceride （TG） and total cholesterol （TC）； quantitative real-time PCR and Western Blot were used to measure the expression levels of related genes and proteins. A one-way analysis of variance was used for comparison between multiple groups， and the Tukey’s test was used for further comparison between two groups. ResultsA total of 32 common targets were obtained for 2，5-DHBA and MAFLD. Within a concentration range of 1.25 — 20 μmol/L， 2，5-DHBA treatment， starting from the concentration of 2.5 μmol/L， increased the viability of AML-12 cells in a dose-dependent manner. The minimum effective concentration of 1.25 μmol/L， the intermediate gradient concentration of 5 μmol/L， and the maximum safe concentration within the effective range of 20 μmol/L were selected for low-， middle-， and high-dose intervention， respectively. Compared with the model group， 2，5-DHBA intervention at concentrations of 1.25， 5， and 20 μmol/L could significantly reduce lipid droplet accumulation in cells （all P<0.05）， and there were significant differences in TG and TC between the model group and the three intervention groups （all P<0.05）. Compared with the control group， the model group had a significant increase in ROS fluorescence intensity （P<0.05）， and compared with the model group， there was a dose-dependent reduction in intracellular ROS level after treatment with 5 μmol/L or 20 μmol/L 2，5-DHBA （P<0.05）. Compared with the model group， 2，5-DHBA treatment at concentrations of 5 and 20 μmol/L significantly downregulated the mRNA expression levels of the lipogenic genes SREBf1 and FASN and upregulated the mRNA expression levels of the key antioxidant gene Nrf2 and its downstream target gene HO-1 （all P<0.05）. Compared with the control group， the expression of p-PI3K/PI3K and p-Akt/Akt in the model group was significantly increased （all P<0.05）. Compared with the model group， the protein expression levels of p-PI3K/PI3K and p-Akt/Akt were significantly decreased after 5 μmol/L 2，5-DHBA treatment （P<0.05）. Compared with the control group， the expression levels of Nrf2 and HO-1 protein in the model group were significantly decreased （all P<0.05）. The protein levels of Nrf2 and HO-1 were significantly increased after treatment with 1.25 μmol/L 2，5-DHBA （P<0.05）. ConclusionNetwork pharmacology and cellular experiments confirm that 2，5-DHBA can alleviate lipid deposition and oxidative stress by regulating the PI3K/Akt signaling pathway， thereby exerting a therapeutic effect on MAFLD.

OBJECTIVE To investigate the ameliorative effect of Weiling decoction on metabolic associated fatty liver disease （MAFLD） based on the gut-liver axis theory.METHODS Male C57BL/6J mice were divided into a blank group （ n ＝9） and a modeling group （ n ＝47）. The modeling group was fed a high-fat diet for 12 weeks to establish the MAFLD model. Forty-five modeled mice were randomly divided into a model group， a positive control group [metformin， 0.2 g/（kg·d）]， and Weiling decoction low-， medium-， and high-dose groups [6.17， 12.34， 24.68 g/（kg·d）， calculated as crude drug]， with 9 mice in each group. Mice in each group received intragastric administration of the corresponding drug solution or normal saline once daily for 4 consecutive weeks. After the last administration， the body weight， liver weight， and liver index were compared among groups； the levels of liver function indicators （alanine transaminase， aspartate transaminase）， lipid indicators （total cholesterol， triglycerides， low-density lipoprotein cholesterol， high-density lipoprotein cholesterol）， inflammatory indicators [tumor necrosis factor-α， interleukin-6 （IL-6）， IL-1β]， oxidative stress indicators （malondialdehyde， superoxide dismutase， glutathione peroxidase）， and intestinal barrier indicators [lipopolysaccharide （LPS）， diamine oxidase， D-lactic acid] were measured； hepatic pathological changes were observed； the expressions of tight junction proteins [zonula occludens-1 （ZO-1）， occludin] in jejunal tissue and proteins related to lipid metabolism [sterol regulatory element-binding protein-1 （SREBP-1）， fatty acid synthase （FASN）， peroxisome proliferator-activated receptor α （PPARα）] and inflammatory pathways [Toll-like receptor 4 （TLR4）， myeloid differentiation primary response protein 88 （MyD88）， nuclear factor-κB p65 （NF-κB p65）] in liver tissue were detected； and changes in gut microbiota were analyzed.RESULTS Compared with the model group， hepatic steatosis and inflammatory injury were significantly alleviated in all Weiling decoction dose groups； body weight （at week 16）， liver weight， liver index， and the levels of liver function indicators， lipid indicators， inflammation indicators， oxidative stress indicators， and intestinal barrier indicators were all generally improved （ P ＜0.05）. The expressions of SREBP-1， FASN， TLR4， and MyD88 and the phosphorylation level of NF-κB p65 in liver tissue were generally decreased or downregulated （ P ＜0.05）， while the expressions of ZO-1 and occludin in jejunal tissue and PPARα in liver tissue were generally increased or upregulated （ P ＜0.05）. Compared with the model group， the Observed species， Chao1， and Ace indices of gut microbiota in the Weiling decoction high-dose group were significantly increased （ P ＜0.05）， and the relative abundances of Verrucomicrobiota， Akkermansia ， etc. were significantly increased， while those of Actinomycetota， Alistipes ， Faecalibaculum ， etc. were significantly decreased （ P ＜0.05）.CONCLUSIONS Weiling decoction can ameliorate MAFLD， and its effect may be associated with regulating gut microbiota， repairing the intestinal mechanical barrier， reducing LPS translocation， thereby inhibiting the TLR4/MyD88/NF-κB inflammatory pathway， alleviating oxidative stress， and regulating SREBP-1/FASN and PPARα-mediated lipid metabolism.

Metabolic dysfunction-associated fatty liver disease （MAFLD） has become the leading chronic liver disease worldwide. As a classic food-medicine homologous Chinese herb， hawthorn fruit has the functions of promoting digestion to resolve food stagnation and activating blood circulation to resolve blood stasis. Modern pharmacological studies have shown that hawthorn fruit and its active components exert diverse pharmacological activities in regulating lipid metabolism， improving insulin resistance， inhibiting inflammatory response， delaying hepatic fibrosis， and protecting hepatocytes. These effects precisely match the “multiple-hit” pathological features of MAFLD， and thus hawthorn fruit shows significant advantages in the prevention and treatment of MAFLD. Drug pair combination is a key strategy in traditional Chinese medicine to enhance efficacy and reduce toxicity. In clinical practice， hawthorn fruit is often combined with Salvia miltiorrhiza， Alisma orientalis， cassia seed， and Poria cocos， and such combination adheres to the principle of “mutual reinforcement and assistance” to enhance efficacy and expand their application in different syndromes of MAFLD. This article systematically reviews the core pharmacological mechanisms of hawthorn fruit in intervening against MAFLD and the research advances in its classic drug combinations， in order to provide a scientific reference for the in-depth development of hawthorn fruit and its combinations and the precise clinical intervention of MAFLD.

Epigenetic mechanisms play a crucial role in the development and progression of nonalcoholic fatty liver disease （NAFLD）， especially among lean individuals. The research on related epigenetic mechanisms has provided new clues and directions for revealing the underlying causes and treatment strategies of NAFLD. This article introduces the role of epigenetics in the development and progression of NAFLD among lean individuals in recent years， analyzes the latest research advances in the epigenetics of NAFLD in this population， and briefly describes the basic concepts of epigenetics， including DNA methylation， histone modifications， and non-coding RNA regulation. This article also discusses how epigenetic alterations impact the pathogenesis， disease progression， and treatment strategies of NAFLD in lean individuals.

OBJECTIVES: With the increasing detection rate of lung nodules, the qualitative problem of lung nodules has become one of the key clinical issues. This study aims to evaluate the value of combining dynamic contrast-enhanced (DCE) MRI based on time-resolved imaging with interleaved stochastic trajectories-volume interpolated breath hold examination (TWIST-VIBE) with T₁ weighted free-breathing star-volumetric interpolated breath hold examination (T₁WI star-VIBE) in identifying benign and malignant lung nodules. METHODS: We retrospectively analyzed 79 adults with undetermined lung nodules before the operation. All nodules of patients included were classified into malignant nodules (n=58) and benign nodules (n=26) based on final diagnosis. The unenhanced T₁WI-VIBE, the contrast-enhanced T₁WI star-VIBE, and the DCE curve based on TWIST-VIBE were performed. The corresponding qualitative [wash-in time, wash-out time, time to peak (TTP), arrival time (AT), positive enhancement integral (PEI)] and quantitative parameters [volume transfer constant (Ktrans), interstitium-to-plasma rate constant (Kep), and fractional extracellular space volume (Ve)] were evaluated. Besides, the diagnostic efficacy (sensitivity and specificity) of enhanced CT and MRI were compared. RESULTS: There were significant differences in unenhanced T₁WI-VIBE hypo-intensity, and type of A, B, C DCE curve type between benign and malignant lung nodules (all P<0.001). Pulmonary malignant nodules had a shorter wash-out time than benign nodules (P=0.001), and the differences of the remaining parameters were not statistically significant (all P>0.05). After T₁WI star-VIBE contrast-enhanced MRI, the image quality was further improved. Compared with enhanced CT scan, the sensitivity (82.76% vs 80.50%) and the specificity (69.23% vs 57.10%) based on MRI were higher than that of CT (both P<0.001). CONCLUSIONS T₁WI star-VIBE and dynamic contrast-enhanced MRI based on TWIST-VIBE were helpful to improve the image resolution and provide more information for clinical differentiation between benign and malignant lung nodules.
Adult ; Humans ; Retrospective Studies ; Magnetic Resonance Imaging ; Plasma ; Tomography, X-Ray Computed ; Lung

Objective To set up a living mice colonoscopy platform to establish an orthotopic model of colorectal cancer in mice under direct vision,and to observe its biological behavior such as metastasis.Methods Eighteen-week-old male C57/BL mice were anesthetized,and the intestinal lumen of the mice was examined by a self-developed living mice colonoscopy and Olympus URF-P5 ureteroscopy,respectively.The imaging effects of the two methods were compared.Human colon cancer HT-29 cells were injected into the colonic mucosa of BALB/c-nu mice under direct vision.The colonoscopy was performed on the 3rd,7th and 15th day after the injection to observe the tumor formation in the intestinal lumen.The mice were sacrificed when the body weight decreased significantly or cachexia appeared,and then the abdominal cavity was examined including the tumor formation and metastasis.Results The self-developed living mice colonoscopy platform can provide clear vision of enteric cavity,and no mice died in the colonoscopy examination.In vivo subcutaneous injection of HT-29 cells in mice was performed with a perforation rate of 15％,a mortality rate of 33.3％,a tumor formation rate of 62.5％,an abdominal metastasis rate of 60％,a liver metastasis rate of 25％,and an abdominal wall transfer rate of 25％.Conclusion The self-developed mice colonoscopy platform can be used for the study of colorectum in living mice.The imaging effect is no less than that of Olympus URF-P5 ureteroscopy.In addition,an orthotopic colorectal cancer model can be established by this platform combing with submucosal injection technology.

Objective To investigate the expression of the Kv3 .4 protein and Kv3 .4 mRNA in various stages of oral carcino‐genesis .Methods The expression of Kv3 .4 protein and Kv3 .4 mRNA were detected by immunohistochemistry (SP method) and RT‐PCR technique respectively in the oral carcinogenesis of SD rat which were induced by 4NQO .Results With the aggravation of the epithelial dysplasia ,Kv3 .4 protein and Kv3 .4 mRNA expression increased gradually .They were strongly positive in oral squa‐mous cell carcinoma .Conclusion The expression of Kv3 .4 protein and Kv3 .4 mRNA levels increased consistently with the aggra‐vation of the epithelial dysplasia .