Both type 2 diabetes mellitus (T2DM) and metabolic associated fatty liver disease (MAFLD) fall within the spectrum of metabolic diseases, and they exhibit a bi-directional causal relationship and robust reciprocal association. Their shared pathological cornerstone is insulin resistance (IR), which involves the interplay of mitochondrial dysfunction and chronic inflammation, forming a cascading pathological process of “IR-mitochondrial dysfunction-inflammation”. This largely explains the notable upward trend in T2DM-MAFLD co-occurrence observed over recent years. Exercise intervention, as a safe and effective non-pharmacological approach, can improve the pathological progression of these patients at multiple levels. Following the logical framework of “pathogenesis-efficacy comparison-molecular mechanisms-clinical translation”, this article systematically compares the efficacy and molecular mechanisms of moderate-intensity continuous training (MICT), resistance exercise, high-intensity interval training (HIIT), and combined training. MICT reduces intrahepatic triglycerides by promoting lipolysis and improving cardiorespiratory fitness; resistance exercise increases muscle mass and basal metabolic rate, offering unique advantages in preserving muscle while reducing fat and improving insulin sensitivity; HIIT is a time-efficient exercise modality that enhances patients’ cardiorespiratory fitness and insulin sensitivity by alternating brief periods of vigorous exertion with recovery periods, with a prominent short-term triglyceride-lowering effect; combined training produces synergistic effects, comprehensively improving glucolipid metabolism and showing the best long-term adherence. Mechanistically, exercise exerts its beneficial effects through three common pathways: (1) AMPK-mediated lipid oxidation and mitochondrial biogenesis; (2) IRS/PI3K/Akt-mediated insulin signaling sensitization; and (3) Nrf2/ARE anti-oxidation and TGF-β/Smads anti-fibrosis regulation. Different exercise modalities activate these pathways with distinct emphases: MICT most directly and persistently activates the AMPK pathway; resistance exercise uniquely improves IRS/PI3K/Akt signaling through muscle mass gain; HIIT induces the highest AMPK activation intensity and triggers unique lactate-mediated signaling regulation; combined training integrates the above multiple mechanistic advantages. For clinical translation, multidisciplinary team collaboration is essential to ensure safety and adherence. Individualized prescriptions should be formulated according to the FITT-VP principle and patient phenotypes—frequency of 3-5 sessions/week of aerobic exercise combined with 2-3 sessions/week of resistance exercise; intensity of moderate-intensity (40%-60% heart rate reserve (HRR))aerobic exercise and 60%-80% of one-repetition maximum (1-RM) for resistance exercise; time of at least 150 min/week of moderate-intensity aerobic exercise, 30-60 min per session; type of combined training as the preferred modality; total volume of≥500-1 000 MET-min/week; and progression adjusted every 4-6 weeks—with real-time adjustments supported by wearable devices, ultimately forming a closed-loop management system from initial assessment to long-term follow-up. Notably, current studies have limitations such as small sample sizes and short intervention periods. Future research should focus on long-term follow-up, multi-omics biomarkers, and combined exercise-drug strategies. In conclusion, the systematic integration of structured, individualized, and sustainable exercise interventions into the multidisciplinary management pathway for patients with T2DM complicated by MAFLD is an urgent need in current clinical practice.

ObjectiveType 2 diabetes mellitus (T2DM) often causes severe hepatic metabolic complications, dominated by metabolic associated fatty liver disease (MAFLD). Persistent hepatic steatosis and oxidative stress further trigger steatohepatitis and progressive liver damage, increasing the mortality risk of diabetic patients. Aerobic exercise effectively improves hepatic lipid metabolism and antioxidant capacity, but poor patient adherence restricts its long-term clinical application. Mitochondrial-derived mitochondrial open reading frame of the 12S rRNA type-c (MOTS-c) is a key peptide regulating insulin sensitivity and hepatic redox homeostasis. This study aimed to explore the protective mechanism of MOTS-c against T2DM-related liver injury and its combined beneficial effect with aerobic exercise via the NRF2/PPARγ signaling axis. This study aimed to investigate whether MOTS-c cooperates with aerobic exercise to alleviate T2DM-associated hepatic oxidative stress and metabolic dysfunction by activating the NRF2/PPARγ axis, and to clarify the molecular and transcriptomic characteristics of their combined intervention. MethodsStable MOTS-c overexpression and knockdown HepG2 cell lines were constructed using lentiviral transfection. An oleic acid-induced cellular lipid accumulation model and NRF2-knockout cell model were applied to verify the NRF2-dependent mechanism ofMOTS-c. Intracellular lipid deposition, triglyceride levels, antioxidant enzyme activities, and the expression of NRF2/PPARγ pathway-related genes and proteins were detected. In vivo, a T2DM rat model with obvious hepatic steatosis was established via a high-fat and high-sucrose diet combined with streptozotocin injection. Model rats received aerobic exercise, MOTS-c intraperitoneal injection, or combined intervention. We detected systemic glycolipid metabolic indicators, hepatic histopathological changes, and the expression of core proteins in the hepatic NRF2/PPARγ axis. Hepatic transcriptomic sequencing was performed to screen differentially expressed genes (DEGs) and enrich key pathways co-regulated by MOTS-c and aerobic exercise. ResultsCellular results showed that MOTS-c overexpression significantly reduced oleic acid-induced lipid deposition, enhanced antioxidant enzyme activity, and upregulated NRF2 and PPARγ expression. Conversely, MOTS-c knockdown aggravated hepatic lipid accumulation and oxidative damage and inhibited NRF2/PPARγ pathway activation. NRF2 knockout completely eliminated the protective effects of MOTS-c on lipid metabolism and redox balance, confirming its NRF2-dependent regulatory mechanism. In T2DM rats, both MOTS-c supplementation and aerobic exercise effectively improved insulin resistance, corrected glycolipid metabolic disorders, and alleviated hepatic steatosis, while consistently activating the hepatic NRF2/PPARγ axis. Compared with single intervention, the combined treatment showed a better improvement trend in hepatic metabolic and oxidative injury, without definitive synergistic effects. Transcriptomic analysis revealed that the co-regulated DEGs of MOTS-c and aerobic exercise were primarily enriched in lipid metabolism and PPAR signaling pathways, with multiple antioxidant and lipid-regulating genes significantly modulated by combined intervention. ConclusionMOTS-c exhibits obvious exercise-mimetic hepatoprotective effects in T2DM. It activates the hepatic NRF2/PPARγ axis to strengthen antioxidant defense, stabilize lipid metabolism, and relieve T2DM-associated hepatic steatosis and oxidative damage. Furthermore, MOTS-c produces additive beneficial effects with aerobic exercise, showing a superior intervention trend on diabetic liver dysfunction. This study identifies the NRF2/PPARγ axis as the core mechanism of MOTS-c-regulated hepatic protection, elucidates the transcriptomic basis of combined intervention, and provides a reliable theoretical basis and potential therapeutic target for clinical intervention in T2DM-complicated MAFLD.

To study the effects of squalene on behavior and related proteins of glutamate toxicity pathways in the mice with chronic unpredictable mild stress (CUMS), thirteen different kinds of CUMS were applied to the male BALB/C mice for 35 days to establish the mouse model of CUMS depression. The stress conditions include food deprivation, noise, stroboscopic lighting, hot stress (45℃), brake, exposure to lower temperature (4℃), shake, soiled cage, clamp tail, water deprivation, swimming, electric shock, presence of a foreign object in the home cage. The mice were treated with squalene at 3 doses (80, 40 and 20 mg·kg^-1·d^-1) through oral administration from the 3rd week continuously. Three weeks later, the impacts were evaluated in the mice with behavioral tests, and malondialdehyde (MDA) and hippocampal glutamate (GLU) contents, the superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity in hippocampus were measured by spectropho-tometry or reversed phase HPLC (RP-HPLC). Western blot was used to examine the expression level of N-methyl-D-aspartate receptor subunits epsilon-2 (NMDAε2), calmodulin kinaseⅡ (CaMKⅡ) and neuronal nitric oxide synthase (NOS1) in hippocampus. Compared with model group, the squalene-treated mice exhibited an increase in body weight, sucrose preference rate and the times of crossing-movement and rearing-movement, shortened the immobility time in the tails suspension test and forced swimming test in the depression mice (P<0.05). Meanwhile, the treated mice had a significant decrease in the contents of GLU and MDA (P<0.05) in hippocampus, increased the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and downregulated the expression of NMDAε2, CaMKⅡ and NOS1 in the hippocampus. In conclusion, squalene shows anti-depressant effect on depressant model in mice, meanwhile the downregulated ROS, related proteins of GLU-NMDAε2-CaMKⅡ-NOS1 signal pathways may be related to the antidepressant effect of squalene.

OBJECTIVETo evaluate the performance of computer-assisted imaging system in the detection of cervical squamous intraepithelial lesion and quality-assurance.

METHODSManual PAP screening (n = 140 580) and image-assisted screening (n = 32 885) were compared for the detection rates of squamous cell abnormalities, the atypical squamous cells (ASC) to squamous intraepithelial lesion (SIL) ratio, the positive rates of high risk human papillomavirus (HR-HPV) test in the case of atypical squamous cells of undetermined significance (ASC-US), and the correlation between cytopathology and histopathology.

RESULTSCompared with manual screening, computer-assisted imaging system showed increased overall positive detection by 0.32%, decreased detection of ASC by 0.21%, increased detection of low-grade squamous intraepithelial lesion (LSIL) by 0.22%, increased detection of high-grade squamous intraepithelial lesion or worse (HSIL) by 0.31%, and decreased ASC to SIL ratio from 2.59 to 1.60. Computer-assisted imaging system did not change the HR-HPV positive rate of the patients who were ASC-US, or the coincidence rate between cytopathology and histopathology. Moreover, the productivity of the laboratory operation increased 58.33%.

CONCLUSIONComputer-assisted imaging system significantly increases the overall positive detection rate of cervical SIL, improves accuracy and work efficiency of screening, decreases the ASC/SIL rate, and strengths the quality-assurance of laboratory testing.

Carcinoma, Squamous Cell ; pathology ; Cervical Intraepithelial Neoplasia ; pathology ; Female ; Humans ; Image Interpretation, Computer-Assisted ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; pathology ; Uterine Cervical Dysplasia ; pathology ; Uterine Cervical Neoplasms ; pathology ; Vaginal Smears ; methods

OBJECTIVETo investigate the effect of 5-FU (5-fluorouracil) on enriching cancer stem cells of HCC cell line BEL-7402 and the biological characteristics of enriched cells.

METHODSThe enriching concentration of 5-FU was determined by CCK-8 (cell counting kit-8). Flow Cytometry was used to determine the changes in cell cycle and positive expression ratio of surface marker CD56, CD54, EpCAM and CD133. The self-renewal and differentiation of positive cells were tested by colony formation assay, and were compared with the control group.

RESULTSEnriching concentration of 5-FU was determined as 10 μg/ml with 48 h incubation. After enrichment, G0/G1 phase cells increased from 57.50 %+/-0.98% to 68.70%+/-3.41% (P<0.05). Whereas S phase cells decreased from 40.26%+/-4.12% to 31.80%+/-4.15% (P<0.01); G2/M phase cells disappeared in experimental group, and was 5.80%+/-1.87% in control group (P<0.01). The proportion of the cell cycle changed with significant statistical differences. Meanwhile, positive rate of cell surface makers CD56, CD54, EpCAM and CD133 increased from 0.57%+/-0.12%, 8.10%+/-6.79%, 0.3%+/-0.01% and 3.20%+/-0.99% to 4.13%+/-0.06%, 50.08%+/-1.69%, 0.55%+/-0.07% and 10.51%+/-1.13%, respectively. The difference was significant (P<0.05). The colony forming ratio of CD56, CD54, EpCAM and CD133 negative cells and positive cells were 2.11%+/-0.21%, 3.32%+/-0.31%; 0.86%+/-0.101%, 2.40%+/-0.52 %; 7.19%+/-0.56%, 7.73%+/-0.71%; 2.70%+/-0.26%, 5.75%+/-0.81%, respectively, and significant differences were found between (P<0.05).

CONCLUSION5-fluorouracil enriched the cancer stem cell population in HCC cell line BEL-7402. CD56 and CD54 can be used as important surface markers in research of liver cancer stem cells.

Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Fluorouracil ; pharmacology ; Humans ; Neoplastic Stem Cells ; cytology ; drug effects ; metabolism

OBJECTIVETo investigate the effect of Changyanqing decoction, a traditional Chinese medicinal preparation, on the expressions of interleukin-10 (IL-10) and intercellular adhesion molecule-1 (ICAM-1) in the colon mucosa of rats with ulcerative colitis.

METHODSThe rats with ulcerative colitis induced by trinitrobenzene sulphonic acid and ethanol enema were randomly divided into 3 groups, namely the model group, sulfasalazine (SASP) group, and Changyanqing decoction group. Daily treatment with intragastric administration and enema of normal saline, SASP (100 mg/kg), and Changyanqing decoction (39.75 mg/kg), respectively, were administered 24 h after the establishment of colitis till the end of the experiment. Another group of rats was used as the normal control group. The disease activity index (DAI) and colon mucosa damage index (CMDI) of the rats were calculated. The activity of myeloperoxidase (MPO) was measured by biochemical method, and the expressions of IL-10 and ICAM-1 protein were measured by ELISA and immunohistochemistry, respectively.

RESULTSCompared with the normal group, the model group showed significantly increased DAI, CMDI, HS score and MPO activity in the colon tissues (P < 0.01), with also significantly increased expression of ICAM-1 (P < 0.01) and decreased expression of IL-10 in the rat colon mucosa (P < 0.01). Treatment with Changyanqing decoction resulted in a significant reduction in DAI, CMDI, HS score and MPO activity (P < 0.01), and decreased the expression of ICAM-1 (P < 0.01) and increased the expression of IL-10 (P < 0.01) in the colon mucosa. The expression of ICAM-1 in the colon mucosa was positively correlated to that of IL-10 (r = 0.927, P < 0.01) and the activity of MPO (r = 0.621, P < 0.01).

CONCLUSIONSChangyanqing decoction has protective effect against rat ulcerative colitis, mediated probably by enhancement of IL-10 expression and reduction in ICAM-1 expression and neutrophil infiltration.

Animals ; Colitis, Ulcerative ; chemically induced ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Interleukin-10 ; biosynthesis ; Intestinal Mucosa ; metabolism ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Trinitrobenzenesulfonic Acid