Objective To investigate the protective effect and underlying mechanism of human umbilical cord mesenchymal stem cell-derived exosomes (hucMSC-Exo) on renal ischemia-reperfusion injury (IRI). Methods hucMSC-Exos were isolated and characterized. A mouse renal IRI model was established and the animals were divided into Sham, IRI, IRI+hucMSC-Exo, IRI+hucMSC-Exo+JY-2 and Sham+JY-2 groups. Serum creatinine (Scr) and blood urea nitrogen (BUN) were measured. Hematoxylin-eosin (HE) staining was used to evaluate renal histopathology. Enzyme-linked immune absorbent assay was performed to determine serum interleukin (IL)-1β and IL-18 levels. Western blotting was used to detect the expression of activating transcription factor 3 (ATF3), Toll-like receptor 4 (TLR4), nuclear factor (NF)-κB, NOD-like receptor protein 3 (NLRP3), cysteineyl aspartate specific proteinase (Caspase)-1 p20 and Gasdermin D(GSDMD). Real-time fluorescent quantitative polymerase chain reaction was employed to measure ATF3, TLR4 and NF-κB messenger RNA (mRNA). Immunohistochemistry was conducted to examine NLRP3, Caspase-1 p20 and GSDMD. An in vitro hypoxia/reoxygenation (H/R) model was established in HK-2 cells and divided into Control, H/R, H/R+hucMSC-Exo, H/R+hucMSC-Exo+JY-2 and Control+JY-2 groups. Western blotting was used to detect the expression of ATF3, TLR4 and NF-κB. Real-time fluorescent quantitative polymerase chain reaction was used to measure NLRP3, GSDMD and Caspase-1 mRNA. Results HucMSC-Exos were successfully isolated and identified. Compared with the Sham group, the IRI group exhibited elevated Scr and BUN, higher tubular injury scores, increased protein expression levels of ATF3, TLR4, NF-κB p65, NLRP3, Caspase-1 p20 and GSDMD, and raised mRNA expression levels of ATF3, TLR4, NF-κB. Compared with the IRI group, the IRI+hucMSC-Exo group showed decreased Scr and BUN, lower tubular injury scores, up-regulated ATF3 protein and mRNA, down-regulated TLR4, NF-κB p65, NLRP3, Caspase-1 p20 and GSDMD protein, and declined TLR4 and NF-κB mRNA. Compared with the IRI+hucMSC-Exo group, the IRI+hucMSC-Exo+JY-2 group exhibited increased Scr and BUN levels, elevated renal tubular injury scores, decreased ATF3 protein expression levels, elevated protein expression levels of TLR4, NF-κB p65, NLRP3, Caspase-1 p20, and GSDMD, decreased ATF3 mRNA expression levels, and elevated mRNA expression levels of TLR4 and NF-κB. (all P < 0.05). Compared with the Control group, the expression levels of ATF3, TLR4 and NF-κB p65 proteins were increased in the H/R group, and the expression levels of NLRP3, Caspase-1 and GSDMD mRNA were increased. Compared with the H/R group, the expression level of ATF3 protein was increased, the expression levels of TLR4 and NF-κB p65 proteins were decreased, and the expression levels of NLRP3, Caspase-1 and GSDMD mRNA were decreased in the H/R+hucMSC-Exo group. Compared with the H/R+hucMSC-Exo group, the expression level of ATF3 protein was decreased, the expression levels of TLR4 and NF-κB p65 proteins were increased, and the expression levels of NLRP3, Caspase-1 and GSDMD mRNA were increased in the H/R+hucMSC-Exo+JY-2 group (all P < 0.05). Conclusions HucMSC-Exos alleviate renal IRI by up-regulating ATF3, thereby negatively regulating the TLR4/NF-κB signaling pathway and subsequently inhibiting pyroptosis.

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.

Schistosomiasis, a zoonotic infectious disease, remains one of the major public health concerns in the world. As a country endemic for schistosomiasis with a long history and substantial disease burdens, China has sustained successive control efforts for over seven decades in the context of the One Health concept, establishing a cross-sectoral collaboration mechanism encompassing health, agriculture, water development resources, and forestry and grassland sectors. A package of interventions have been employed tailored to management of the source of Schistosoma japonicum infections, Oncomelania hupensis snail control, improved water supply and sanitation, environmental modification, and risk surveillance, and a “human-animal-environment” collaborative prevention and control model has been gradually generated. As a consequence, the transmission of schistosomiasis was interrupted across China in 2023. To achieve the goal of schistosomiasis elimination and sustainably consolidate the achievements of elimination in China, it is still necessary to enhance the long-term cross-sectoral coordination mechanism, promote interdisciplinary collaboration for innovating control technologies and strategies, establish an intelligent surveillance network covering the entire region, and strengthen the response and emergency management capabilities. In addition, distilling China’s schistosomiasis control and elimination experiences will provide a “One Health” model for the prevention and control of other parasitic and tropical diseases, thereby contributing to global disease control and sustainable development.

ObjectiveThis study aims to investigate the regulatory effects of microcapsules with different matrix stiffness on the survival, proliferation, adhesion, and osteogenic differentiation of mesenchymal stem cells (MSCs), to further explore the relationship between matrix stiffness and MSCs osteogenic behavior, and to evaluate the in vivo histological responses and osteogenic-related protein expression of the microcapsules using an animal subcutaneous implantation model. MethodsIn this study, three-dimensional carbon nanofiber (CNF)/sodium alginate (SA) microcapsules with different matrix stiffness were constructed. By adjusting the CNF content, three groups of microcapsules with stiffness values of 27.05, 35.90, and 52.40 kPa were prepared, and their physicochemical properties, including morphology, mechanical properties, particle size, chemical structure, and degradation properties, were characterized. The effects of these microcapsules with different stiffness on the survival, adhesion, and osteogenic differentiation of MSCs were evaluated. Live/dead staining was used to observe cell survival and distribution in the microcapsules, while alkaline phosphatase (ALP) activity and the expression of osteopontin (OPN) and osteocalcin (OCN) were examined to assess osteogenic differentiation. Immunofluorescence staining was further performed to observe the expression of Yes-associated protein (YAP) and integrin β1 under different stiffness conditions. In addition, a rat subcutaneous implantation model was used to evaluate the in vivo tissue compatibility of the microcapsules. Histological staining and immunohistochemical analysis were carried out to examine local tissue responses and the expression of osteogenic-related proteins. ResultsThe results showed that all three groups of microcapsules exhibited regular and uniform morphology and similar particle size, and all groups showed good degradability. Except for the differences in mechanical properties, no obvious differences were found in the other physicochemical properties among the three groups. Live/dead staining indicated that all groups of microcapsules had good cytocompatibility. MSCs in the low-stiffness group were mostly round, whereas cells in the medium- and high-stiffness groups showed more obvious spreading. Higher matrix stiffness was more favorable for MSC adhesion, growth, and survival. ALP staining and quantitative analysis showed that ALP activity increased significantly with increasing matrix stiffness. The expression of the osteogenic-related proteins OPN and OCN also increased with increasing stiffness, and the high-stiffness group showed the strongest positive signals. Immunofluorescence results further indicated that YAP and integrin β1 fluorescence signals tended to increase in MSCs with increasing matrix stiffness, suggesting that matrix stiffness may influence MSC osteogenic differentiation through stiffness-related cell adhesion and mechanotransduction. In vivo experiments showed that all three groups of microcapsules exhibited good histocompatibility after subcutaneous implantation, although different degrees of degradation and local tissue response were observed. Immunohistochemical analysis showed that the expression levels of OPN and OCN increased with increasing matrix stiffness, and the high-stiffness group exhibited higher expression of these osteogenic-related proteins. ConclusionCNF endows SA microcapsules with tunable matrix stiffness. While maintaining good biocompatibility, the high-stiffness microenvironment can effectively promote the adhesion and osteogenic differentiation of MSCs, accompanied by increased ALP activity and enhanced OPN and OCN expression. The increased YAP and integrin β1 fluorescence signals in the high-stiffness group further indicate that matrix stiffness is closely related to cellular mechanical responses. These findings provide a three-dimensional cell carrier with potential application value for MSC delivery and bone tissue regeneration based on mechanical microenvironment regulation.

ObjectiveTo investigate the heterogeneity of PD-L1 expression profiles between primary tumors and paired metastatic lymph nodes in advanced non-small cell lung cancer （NSCLC）， and to determine the predictive value of key immune marker expression for the efficacy of first-line targeted therapy and chemotherapy. MethodsA retrospective analysis was conducted on 128 patients with histologically confirmed NSCLC and corresponding metastatic lymph nodes， who received first-line tyrosine kinase inhibitors （TKIs） or platinum-based doublet chemotherapy at Guangdong Provincial People's Hospital between April 2017 and February 2020. Immunohistochemistry （IHC） was employed to evaluate PD-L1 expression in primary tumors and metastatic lymph nodes. The Wilcoxon signed-rank test was performed to compare PD-L1 expression characteristics between primary tumors and metastatic lymph nodes. Kaplan-Meier survival curves were constructed， and the Log-rank test was used to compare differences between groups. ResultsAmong 28 paired cases， PD-L1 expression levels were numerically higher in metastatic lymph nodes than in primary NSCLC tumors （median：32.5 vs. 10.0）， although the difference did not reach statistical significance （MD = 5.000， P = 0.083）. Survival analysis revealed that in patients with driver gene-positive NSCLC， high PD-L1 expression （TPS ≥ 50%） in lymph nodes was significantly associated with shorter progression-free survival （PFS） following first-line TKI therapy ［median PFS：4.0（TPS≥50%） vs. 8.9（1%≤TPS＜50%） vs. 18.0 （TPS＜1%） months， χ²=15.284， P＜0.001］. Conversely， in patients with driver gene-negative NSCLC， high PD-L1 expression in lymph nodes was associated with longer PFS ［median PFS：7.9 （TPS≥50%） vs. 3.0 （1%≤TPS＜50%）months， χ²=8.436， P=0.004］ and overall survival （OS） ［median OS：28.8 （TPS≥50%） vs. 14.2 （ 1%≤TPS＜50%） months， χ²=4.010， P=0.045］ after first-line chemotherapy. ConclusionPD-L1 expression in metastatic lymph nodes is largely consistent with that in primary tumors of patients with advanced NSCLC. High PD-L1 expression in lymph nodes is associated with improved survival outcomes in driver gene-negative patients undergoing chemotherapy， whereas it portends a poor prognosis with TKI treatment in driver gene-positive patients. These findings support the application of PD-L1 expression levels in metastatic lymph nodes to guide personalized precision medicine strategies for advanced NSCLC.

In human assisted reproductive technology (ART), embryos experience an extremely high rate of developmental arrest (over 50%) during the 5-day developmental window prior to blastocyst formation, which constitutes a major bottleneck limiting clinical pregnancy rates. A recent paper published in Cell titled “Two distinct causes contribute to the low efficiency of human pre-implantation development” reveals the two independent causes underlying this phenomenon. Utilizing a custom-developed high-throughput dual-view live-cell light-sheet microscope, the research team achieved the first continuous live-cell imaging of human and non-human primate embryos with high spatiotemporal resolution for up to 120 h. The study discovered that embryos are highly prone to errors during the second mitosis. Multipolar spindle assembly, triggered by centriole overduplication, leads to chromosome segregation errors and micronuclei formation, which subsequently induces early developmental arrest (prior to the 8-cell stage). Conversely, late developmental arrest (on day 4) is primarily driven by the endoplasmic reticulum (ER) stress response. Furthermore, the study demonstrated that transient treatment with a PLK4 inhibitor (centrinone) can effectively prevent spindle defects at the 2-cell stage. This research not only reshapes our understanding of the developmental dynamics of human pre-implantation embryos but also provides new insights into the mechanisms underlying human pre-implantation developmental arrest and offers valuable references for future studies on assisted reproduction and potential intervention strategies.

OBJECTIVES: To investigate the effects of dihydroartemisinin (DHA) combined with doxorubicin (DOX) on proliferation and apoptosis of triple-negative breast cancer cells and explore the underlying molecular mechanism. METHODS: MDA-MB-231 cells were treated with 50, 100 or 150 μmol/L DHA, 0.5 μmol/L DOX, or with 50 μmol/L DHA combined with 0.5 μmol/L DOX. The changes in proliferation and survival of the treated cells were examined with MTT assay and colony-forming assay, and cell apoptosis was analyzed with flow cytometry. Western blotting was performed to detect the changes in protein expression levels of PCNA, cleaved PARP, Bcl-2, Bax, STAT3, p-STAT3, HIF-1α and survivin. RESULTS: The IC₅₀ of DHA was 131.37±29.87 μmol/L in MDA-MB-231 cells. The cells with the combined treatment with DHA and DOX showed significant suppression of cell proliferation. Treatment with DHA alone induced apoptosis of MDA-MB-231 cells in a dose-dependent manner, but the combined treatment produced a much stronger apoptosis-inducing effect than both DHA and DOX alone. DHA at 150 μmol/L significantly inhibited clone formation of MDA-MB-231 cells, markedly reduced cellular expression levels of PCNA, p-STAT3, HIF-1α and survivin proteins, and obviously increased the expression level of cleaved PARP protein and the Bax/Bcl-2 ratio, and the combined treatment further reduced the expression level of p-STAT3 protein and increased the Bax/Bcl-2 ratio. CONCLUSIONS DHA combined with DOX produces significantly enhanced effects for inhibiting cell proliferation and inducing apoptosis in MDA-MB-231 cells possibly as result of DHA-mediated negative regulation of the STAT3/HIF-1α pathway.
Humans ; STAT3 Transcription Factor/metabolism* ; Apoptosis/drug effects* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Doxorubicin/pharmacology* ; Triple Negative Breast Neoplasms/metabolism* ; Cell Line, Tumor ; Artemisinins/pharmacology* ; Female ; Cell Proliferation/drug effects* ; Signal Transduction/drug effects* ; Survivin

The liver, skeletal muscle, and adipose tissue are central energy-metabolizing organs and insulin-sensitive tissues, playing a crucial role in maintaining glucose homeostasis. As the powerhouse of the cell, mitochondria not only regulate insulin secretion but also oversee the oxidative phosphorylation and β-oxidation of fatty acids, processes vital for the metabolism of carbohydrates and fats, as well as the synthesis of ATP. The mitochondrial quality control system is of paramount importance for sustaining mitochondrial homeostasis, achieved through mechanisms such as protein homeostasis, mitochondrial dynamics, mitophagy, and biogenesis. Evidence suggests that dysfunctional mitochondria may significantly contribute to insulin resistance and ectopic fat storage in the liver, offering new insights into the strong correlation between mitochondrial dysfunction and the development of obesity, diabetes mellitus type 2 (T2DM), and non-alcoholic fatty liver disease (NAFLD). This manuscript aims to delve into the precise mechanisms by which imbalances in mitochondrial quality control lead to metabolic disorders in the liver, skeletal muscle, and adipose tissue, the 3 major insulin-sensitive organs. In the liver, mitochondrial dysfunction can lead to disturbances in glucose and lipid metabolism, resulting in insulin resistance and fat accumulation—a key factor in the development of NAFLD. In skeletal muscle, reduced mitochondrial function can decrease ATP production, weakening the muscle’s ability to uptake glucose, thereby exacerbating insulin resistance. In adipose tissue, mitochondrial dysfunction can impair adipocyte function, leading to lipotoxicity and inflammatory responses,which further contribute to insulin resistance and the onset of metabolic syndrome. Moreover, the interorgan crosstalk among these 3 tissues is essential for overall metabolic homeostasis. For instance, hepatic gluconeogenesis and glucose utilization in skeletal muscle are both influenced by the health status of their respective mitochondrial populations. The conversion between different types of adipose tissue and the ability to store lipids depend on normal mitochondrial function to avert ectopic fat accumulation in other organs. In summary, this manuscript emphasizes the critical role of mitochondrial quality control in maintaining the metabolic stability of the liver, skeletal muscle, and adipose tissue. It elucidates the specific mechanisms by which mitochondrial dysfunction in these organs contributes to the development of metabolic diseases, providing a foundation for future research and the development of therapeutic strategies targeting mitochondrial dysfunction.

AIM:To investigate the effect of silent information regulator 1(SIRT1)on the degree of aging and apoptosis in a mouse cardiomyocyte aging model through the regulation of Wnt/β-catenin pathway.METHODS:An in vi-tro aging model was established by inducing HL-1 cells with 40 μmol/L D-galactose(D-Gal).The HL-1 cells were trans-fected with a lentivirus overexpressing SIRT1,and the transfection efficiency was verified by Western blot.Western blot was used to detect the protein expression levels of SIRT1,P53,P21,cleaved caspase-3,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),β-catenin,Wnt3a and c-Myc.Senescence-associated β-galactosidase(SA-β-Gal)staining was used to detect cellular senescence level.MTT colorimetric assay was used to detect the cell viability,and flow cytometry was used to detect the apoptosis.RESULTS:Treatment of HL-1 mouse cardiomyocytes with D-Gal led to in-creases in the expression levels of aging-related proteins P53 and P21,as well as an increase in SIRT1 protein level.Addi-tionally,the SA-β-Gal staining showed a significant increase in the positive area(P＜0.05).The expression levels of apop-tosis-related proteins cleaved caspase-3 and Bax were elevated,while the level of the anti-apoptotic protein Bcl-2 was re-duced(P＜0.05).There was a marked decrease in cell viability(P＜0.05),and flow cytometry analysis demonstrated a significant increase in cell apoptosis rate(P＜0.05),which was positively correlated with the duration of D-Gal treatment.Overexpression of SIRT1 notably reduced both aging and apoptosis levels after 48 h of D-Gal treatment(P＜0.05).After D-Gal treatment,the expression levels of β-catenin,c-Myc and Wnt3a proteins were up-regulated.However,these levels were reduced when SIRT1 was overexpressed.Moreover,the addition of LiCl,a Wnt/β-catenin pathway agonist,resulted in increased expression levels of β-catenin,c-Myc and Wnt3a proteins compared with the group with SIRT1 overexpres-sion and D-Gal treatment(P＜0.05).CONCLUSION:SIRT1 inhibits cardiomyocyte apoptosis and alleviates cardiomyo-cyte aging through the Wnt/β-catenin pathway.

Objective To investigate the correlation between apathy and imaging markers in pa-tients with aCSVD.Methods A total of 143 patients diagnosed with aCSVD and hospitalized in the Department of Neurology of the First Affiliated Hospital of Baotou Medical College,Inner Mongolia University of Science and Technology from August 2023 to August 2024 were continu-ously included as the study objects.According to MAES,they were divided into an apathetic group(MAES score＞14,68 cases)and a non-apathetic group(MAES score ≤14,75 cases).The clinical data and imaging markers were compared between the two groups.Results The apathetic group had significantly older age and larger ratio of hypertension,but shorter years of education and lower MAES score than the non-apathetic group(P＜0.05,P＜0.01).The apathetic group also had notably higher Fazekas score of white matter hyperintensity(WMH),larger recent small sub-cortical infarct(RSSI),lacunar infarct(LI),and perivascular space(PVS)in the basal ganglia and the centrum semiovale,more obvious cerebral atrophy and cerebral microbleed(CMB),and high-er total imaging burden score when compared with the non-apathetic group(P＜0.01).In the aCSVD patients,the MAES score was positively correlated with WMH Fazekas score,RSSI,LI,basal ganglia PVS,centrum semiovale PVS,cerebral atrophy,CMB,and total imaging burden score(P＜0.01).WMH Fazekas score was an independent risk factor for apathy in the aCSVD patients(OR=2.218,95％CI:1.343-3.664,P=0.002).Conclusion The higher the score of ima-ging markers in patients with aCSVD,the more severe the apathy.