1.Association between the incidence of hemorrhagic fever with renal syndrome and meteorological factors in Shenzhen City from 2012 to 2019
Liangqiang LIN ; Dongfeng KONG ; Lanbin XIANG ; Zhigao CHEN ; Yanmin QIN ; Yuefa ZHUANG ; Yang LIU ; Jianfeng LI
Chinese Journal of Schistosomiasis Control 2026;38(2):194-199
Objective To examine the association between epidemiological characteristics of hemorrhagic fever with renal syndrome (HFRS) and meteorological factors in Shenzhen City during the period from 2012 to 2019. Methods Average atmospheric pressure, average air temperature, average relative humidity, precipitation, wind speed, and sunshine duration were captured from Meteorological Bureau of Shenzhen City each month from 2012 to 2019. The average monthly rodent densities in Shenzhen City from 2012 to 2019 were acquired from the Vector Surveillance Management System of Guangdong Provincial Center for Disease Control and Prevention, and the monthly HFRS incidence was retrieved from Shenzhen Municipal Disease Surveillance System from 2012 to 2019. The correlation between meteorological factors and the monthly incidence of HFRS was examined us ing Spearman’s rank correlation in Shenzhen City, and the temporal trends in monthly HFRS incidence and the degrees of freedom for the rodent density were determined in Shenzhen City with a generalized additive model. The optimal lag time was identified using excess risk (ER) and its 95% confidence interval (CI), and univariate and multivariate models were fitted to evaluate the impact of meteorological factors on HFRS incidence in Shenzhen City. Results The median number of incident HFRS cases was 3.00 (interquartile range, 3.25) in Shenzhen City from 2012 to 2019, with an average air temperature of (23.44 ± 4.91) °C, average relative humidity of (76.05 ± 7.61)%, median precipitation of 4.10 (interquartile range, 6.83) mm, average wind speed of (1.97 ± 0.26) m/s, average sunshine duration of (5.17 ± 1.64) h, and median monthly rodent density of 1.74% (interquartile range, 2.52%). Spearman’s rank correlation analysis showed that the average air temperature positively correlated with average relative humidity (rs = 0.420, P < 0.05), precipitation (rs = 0.658, P < 0.05) and sunshine duration (rs = 0.633, P < 0.05), and the atmospheric pressure negatively correlated with average air temperature (rs = −0.925, P < 0.05), relative humidity (rs = −0.614, P < 0.05), precipitation (rs = −0.789, P < 0.05) and sunshine duration (rs = −0.437, P < 0.05), while the average relative humidity correlated positively with precipitation (rs = 0.724, P < 0.05) and negatively with sunshine duration (rs = −0.218, P < 0.05). Univariate modeling analysis showed that the ERs and their 95% CI were 0.639% (0.540%, 0.737%) for average atmospheric pressure, −7.157% (−8.113%, −6.190%) for average air temperature, −3.603% (−4.219%, −2.985%) for average relative humidity, −5.889% (−7.085%, −4.669%) forprecipitation,21.881% (−5.149%, 56.612%) for average wind speed, and −13.877% (−16.641%, −11.022%) for sunshine duation (all P values < 0.05). Multivariate modeling analysis showed that in the ensemble model combining average atmospheric pressure and precipitation, the highest ER (6.686%) was caused by increased average atmospheric pressure, and the highest absolute ER values for average air temperature (6.615%), average relative humidity (3.107%) and precipitation (5.386%) were seen after adjustment only for sunshine duration (all P values < 0.05), while the highest absolute ER for sunshine duration (11.875%) was found after adjustment for precipitation (P < 0.05). Conclusions An increase in average air temperature, relative humidity, precipitation and sunshine duration resulted in a reduced incidence rate of HFRS in Shenzhen City from 2012 to 2019, and an increase in average atmospheric pressure increased the incidence of HFRS. Meteorological factors are important determinants affecting HFRS incidence in Shenzhen City.
2.Systematic review of the relationship between dynamic changes in inflammatory markers and therapeutic efficacy in NSCLC patients during EGFR-TKIs therapy
Liying WU ; Jingyi YANG ; Yating ZENG ; Ling YONG ; Weifeng SHAO ; Wei LIU
China Pharmacy 2026;37(11):1490-1495
OBJECTIVE To systematically evaluate the correlation between dynamic changes in inflammatory markers during treatment with epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in non-small cell lung cancer (NSCLC) patients and therapeutic efficacy, with the aim of providing evidence-based support for clinical prognosis assessment and treatment strategy adjustment. METHODS Databases including PubMed, Embase, Cochrane Library, CNKI, Wanfang Data, and CBM were searched from the inception to July 20, 2025. Following literature screening, data extraction and quality assessment, descriptive analysis was conducted on the outcomes of included studies. RESULTS A total of eight studies were included to analyze the correlation of 6 inflammatory markers before and after treatment with EGFR-TKIs with therapeutic efficacy. The risk of bias assessment identified six high-quality studies and two moderate-quality studies. Among these studies, seven studies demonstrated that lower levels of neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), platelet-to-lymphocyte ratio (PLR), and monocyte-to-lymphocyte ratio (MLR), higher lymphocyte-to-monocyte ratio (LMR) before treatment, as well as decreased NLR and MLR and increased LMR after treatment were associated with longer median progression-free survival. Five studies indicated that lower levels of NLR, dNLR, PLR, and interleukin-6 (IL-6), higher LMR before treatment as well as decreased NLR and dNLR and increased LMR were associated with longer median overall survival. Three studies indicated that lower levels of IL-6 were associated with a higher objective response rate, while the association of these markers after treatment remained controversial; another study showed that an early decline in NLR, MLR, and PLR after treatment may be associated with objective response benefit. CONCLUSIONS Lower inflammatory levels during EGFR-TKIs therapy correlate with better therapeutic efficacy in NSCLC patients.
3.Research progress on the clinical application and mechanism of Chaihu jia longgu muli decoction in the treatment of neurological diseases
Yimeng KONG ; Zhiyong LIU ; Chengsi DUAN ; Lanyun SHI ; Meina ZHAO ; Mengyuan ZHANG ; Chen YANG
China Pharmacy 2026;37(11):1515-1520
Neurological diseases have high morbidity and disability rates, posing a severe threat to human health. Cli nical manifestations include motor, sensory, cognitive and conscious disorders. Chaihu jia longgu muli decoction is derived from Treatise on Febrile Diseases , with the effects of harmonizing Shaoyang, activating Yang and clearing heat, and tranquilizing the mind. This paper systematically reviews the research progress in clinical application and mechanism of Chaihu jia longgu muli decoction in the field of neurological diseases. It has been found that the decoction shows favorable efficacy in various neurological diseases such as insomnia, depression, epilepsy, vertigo, migraine and vascular dementia. The specific mechanisms are related to regulating neurotransmitter levels, repairing neuronal function, alleviating neuroinflammation, improving mitochondrial dysfunction and regulating intestinal flora. In the future, standardized prospective follow-up cohorts should be established, and core outcome indicators should be clearly defined to strengthen the evidence base. Furthermore, multidisciplinary research should be leveraged to expand the therapeutic value of Chaihu jia longgu muli decoction in the management of neurological diseases.
4.Intravitreal Conbercept for macular edema secondary to non-ischemic retinal vein occlusion
Min YANG ; Shanshan LI ; Shuang LIU ; Dawei ZHANG
International Eye Science 2026;26(7):1147-1151
AIM:To observe the clinical efficacy of intravitreal injection of conbercept in the treatment of macular edema secondary to non-ischemic retinal vein occlusion(RVO).METHODS: Single center retrospective study. ME patients secondary to non-ischemic RVO admitted to the hospital from January 2023 to March 2024 were selected, and were divided into central retinal vein occlusion(CRVO)group and branch retinal vein occlusion(BRVO)group according to the location of obstruction. All patients were treated with intravitreal injection of conbercept once a month for 3 mo. The best corrected visual acuity(BCVA), macular foveal thickness(CMT), superficial capillary density(SVD), deep capillary density(DVD), and foveal avascular zone(FAZ)area were recorded before and after treatment(with 3 injections per course)at 1, 3, and 6 mo.RESULTS:This study included a total of 120 ME secondary to non-ischemic RVO patients(128 eyes), who were divided into CRVO group(51 cases, 56 eyes, 31 males, 20 females, mean age 61.39±10.32 y)and BRVO group(69 cases, 72 eyes, 41 males, 28 females, mean age 61.48±10.41 y)based on the location of obstruction. There was no significant difference in general data between the two groups before treatment(both P>0.05). After 1, 3, and 6 mo of treatment, both groups showed improvement in BCVA, CMT, SVD, and DVD compared to before treatment(all P<0.05). BCVA in the BRVO group was better than that in the CRVO group at all time points after treatment(all P<0.05), while there was no difference in CMT, SVD, and DVD between the two groups(all P>0.05); There was no significant difference in FAZ area between the two groups before and after treatment(both P>0.05). Follow up for 6 mo showed no significant difference in the incidence of complications between the two groups of patients(both P>0.05), but there was a significant difference in the recurrence rate(P<0.05).CONCLUSION: The first intravitreal injection of conbercept is effective in treating macular edema caused by non-ischemic CRVO and BRVO, improving visual function, reducing macular edema, and repairing retinal structure and blood flow perfusion. Notably, the recovery of visual function and improvement of capillary density are more significant in BRVO patients.
5.Relationship between serum Sirt1-6 levels and oxidative stress indicators in age-related cataract patients and their diagnostic value
Lu LU ; Fangfang WU ; Wenwen WU ; Lidong YANG ; Yawen LIU
International Eye Science 2026;26(7):1239-1245
AIM: To investigate the relationship between serum levels of sirtuins(Sirts)1-6 and oxidative stress markers, and to evaluate their diagnostic value for age-related cataract(ARC).METHODS:A prospective cohort of ARC patients admitted to the hospital between June 2019 and July 2021 was enrolled as the study subjects. Concurrently, age-matched healthy individuals undergoing routine physical examinations at the hospital during the same period were recruited as the control group. Baseline demographic and clinical characteristics were compared between the two groups. Serum levels of Sirt1-6 and oxidative stress markers including malondialdehyde(MDA), superoxide dismutase(SOD), glutathione peroxidase(GSH-Px), and total antioxidant capacity(TAC)were compared. Pearson correlation analysis was performed to assess the associations between serum Sirt1-6 levels and oxidative stress parameters. Multivariate Logistic regression analysis was conducted to analysis risk factors associated with ARC occurrence. Receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of serum Sirt1-6 levels and oxidative stress indicators for ARC.RESULTS: The study included 127 ARC patients, comprising 68 men and 59 women, with a mean age of 66.05±2.58 y; the concurrent control group consisted of 121 patients, comprising 63 men and 58 women, with a mean age of 65.54±2.86 y. Serum levels of Sirt1, 3 and 6, SOD, GSH-Px, and TAC were significantly lower in the ARC group compared to the control group(all P<0.001), whereas MDA levels were markedly elevated(P<0.001). Pearson correlation analysis revealed that serum levels of Sirt1, Sirt3, Sirt6 in the ARC group were positively correlated with SOD, GSH-Px, and TAC, and negatively correlated with MDA. Multivariate Logistic regression analysis demonstrated that serum Sirt1, Sirt6, MDA, SOD, and GSH-Px were significantly associated with the occurrence of ARC(all P<0.001). ROC curve analysis showed that the combination of Sirt1, Sirt6, MDA, SOD, and GSH-Px yielded an area under the curve(AUC)of 0.995 for diagnosing ARC, which was significantly higher than that of Sirt1 alone(Z=4.978,P<0.001), Sirt6 alone(Z=7.487,P<0.001), MDA alone(Z=6.449,P<0.001), SOD alone(Z=5.773,P<0.001), or GSH-Px alone(Z=5.056,P<0.001), indicating superior diagnostic accuracy of the multimarker panel(P<0.05). CONCLUSION: Serum levels of Sirt1, 3, and 6 are generally reduced in ARC patients and are closely associated with oxidative stress imbalance. Specifically, decreased serum levels of Sirt1 and Sirt6, along with oxidative stress markers(elevated MDA and reduced SOD and GSH-Px)are identified as risk factors for ARC. Moreover, the combined detection of these indicators presents high diagnostic value for ARC.
6.Teprotumumab combined with glucocorticoid pulse therapy for thyroid-associated ophthalmopathy
Yuan LIU ; Qian YANG ; Juan DU ; Hu CHANG ; Ge GAO
International Eye Science 2026;26(7):1264-1269
AIM: To explore the clinical therapeutic effect of teprotumumab combined with glucocorticoid pulse therapy for thyroid-associated ophthalmopathy(TAO), and its impacts on thyroid function, levels of inflammatory factors, and adverse reactions in patients. METHODS: Active TAO patients admitted to the Ophthalmology Department were enrolled and randomly divide into the steroid group and the combined group. Then the steroid group was treated with glucocorticoid pulse therapy, while the combined group was combined with intravenous infusion of teprotumumab on the basis of the steroid group. The clinical therapeutic effect, the CAS, OSDI, M-C-TAO-QOL scores, ocular sign indicators(fissure width, proptosis), levels of inflammatory factors(TNF-α, CRP, IL-17), thyroid function(TSH, FT3, FT4)before and after treatment, and occurrence of adverse reactions were compared between two groups.RESULTS:Totally 96 TAO patients(192 eyes)were included, with 48 cases(96 eyes)in each group. In the combined group, there were 17 males and 31 females, with an average age of 51.85±3.53 y; in the steroid group, there were 14 males and 34 females, with an average age of 51.26±3.84 y. The total effective rate of the combined group(94%)was higher than that of the steroid group(79%)(P<0.05). After treatment, the CAS score, OSDI score, fissure width, proptosis, levels of TNF-α, CRP, and IL-17 in the combined group were all lower than those in the steroid group, and the M-C-TAO-QOL score was higher than that in the steroid group(P<0.05). However, there was no difference in thyroid function indicators and adverse reactions between two groups after treatment(P>0.05). CONCLUSION: The combination of teprotumumab and glucocorticoid pulse therapy for TAO has a prominent therapeutic effect. Meantime, it can more effectively control ocular inflammation, improve ocular signs and quality of life of patients, and has no obvious adverse effect on thyroid function, with controllable safety.
7.Wisdom Inheritance of Distinguished Physicians' Experience Through Integration of Multimodal Data and AIGC: A Case Study on Experience in Diagnosis and Treatment of Lung Cancer with Phlegm-dampness and Blood Stasis Syndrome by Distinguished Traditional Chinese Medicine Physicians of Sichuan School
Yang YU ; Yadong MU ; Wenping LIU ; Chongcheng XI ; Li ZHANG ; Yan GAO ; Cen JIANG ; Quansheng FENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):14-25
Lung cancer, with persistently high incidence and mortality rates, remains a significant global health challenge. By taking the study on the experience in diagnosis and treatment of lung cancer with phlegm-dampness and blood stasis syndrome by distinguished traditional Chinese medicine physicians of the Sichuan School as an example, the diagnosis and treatment system for lung cancer with phlegm-dampness and blood stasis syndrome, which was formed in response to the humid and foggy environment of the Sichuan Basin, possesses unique value. However, traditional inheritance modes face challenges such as fragmentation, lack of standardization, and insufficient quantification, which hinder the promotion and application of this experience. This research focused on how to leverage multimodal data and artificial intelligence-generated content (AIGC) to achieve precise analysis, intelligent inheritance, and clinical innovation of the experience in diagnosis and treatment of lung cancer with phlegm-dampness and blood stasis syndrome by distinguished traditional Chinese medicine physicians of the Sichuan School. By integrating multimodal data (encompassing four diagnostic methods of traditional Chinese medicine, modern medical imaging, clinical laboratory tests, molecular biology, and regional environmental information), a precise diagnosis and treatment system integrating macro and micro perspectives for the "disease, syndrome, and pathogenesis" was constructed. The research yielded the following results: (1) In precise syndrome differentiation, the objective quantification of the phlegm-dampness and blood stasis syndrome was achieved. By constructing a "four diagnostic methods, imaging, and molecule" correlation model, the study revealed intrinsic links between tongue and pulse parameters and the tumor microenvironment, as well as between regional climatic factors and syndrome characteristics, enabling real-time dynamic monitoring of efficacy. (2) In elucidating patterns, the study systematically explored the syndrome differentiation thoughts of Sichuan School physicians, such as the timing of purgation and tonification. A "pathogenesis, syndrome complex, and prescriptions and herb" network model was constructed, which accurately elucidated the synergistic action mechanisms of core herb pairs and quantified the dynamic compatibility patterns of reinforcing healthy Qi and eliminating pathogenic factors. (3) In intelligent empowerment, an auxiliary system integrating intelligent syndrome differentiation, treatment plan generation, and efficacy evaluation was built. This system can fuse regional characteristics with individual data, dynamically generate and optimize personalized prescriptions aligned with the experience of Sichuan School, and predict efficacy trends and potential adverse reactions. The integration of multimodal data and AIGC can effectively facilitate the structured inheritance and clinical translation of distinguished physicians' experience. The established intelligent diagnosis and treatment model integrating traditional Chinese medicine and Western medicine demonstrates clear potential in prolonging patients' progression-free survival, alleviating symptoms, and reducing adverse reactions to treatment. This study provides a referential methodological framework for the traditional Chinese medicine experience in diagnosis and treatment of lung cancer, especially the empirical inheritance and modernized development of regional academic schools. It contributes to advancing clinical diagnosis and treatment toward greater precision and personalization.
8.Action of Immune Microenvironment and Metabolic Reprogramming in Hepatocellular Carcinoma Based on "Deficiency of Healthy Qi and Stasis Toxins"
Xia LI ; Jiexiong YANG ; Xiyang LIU ; Wenjun WU ; Cen JIANG ; Quansheng FENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):100-109
Hepatocellular carcinoma (HCC), a malignancy with high mortality, exhibits poor survival rates and prognosis. The profound suppression of the tumor immune microenvironment (TIME) and the abnormal hyperactivity of metabolic reprogramming (MR) are the two primary factors driving HCC progression. Traditional Chinese medicine has demonstrated significant efficacy in HCC treatment. The team proposed that "deficiency of healthy Qi and stasis toxins" was the core pathogenesis of HCC, closely associated with TIME suppression and MR hyperactivity. This paper proposed that a suppressed state of the TIME was the biological manifestation of "deficiency of healthy Qi", where the functional exhaustion of effector T lymphocytes and natural killer cells reflected the decline of "healthy Qi" in eliminating pathogens. Conversely, the expansion and activation of immunosuppressive cells, such as regulatory T cells (Tregs), M2-like tumor-associated macrophages (TAM-M2), and myeloid-derived suppressor cells (MDSCs) , represent the dysfunction of "healthy Qi" in maintaining homeostasis. MR serves as the material basis of "stasis toxins". Stasis toxins exhibit heat stagnation, manifested by abnormal hyperactivity of glycolysis and lipid synthesis. They demonstrate migratory propensity, as toxic metabolites like lactic acid and prostaglandin E2 promote tumor invasion and metastasis. They display a consumptive nature, reflected in the functional suppression of immune cells. The vicious cycle between TIME and MR is the biopathological reflection of "deficiency of healthy Qi intertwined with stasis toxins". Immunosuppression exacerbates MR, while toxic metabolites further impair immune function, establishing a pathogenic chain of "deficiency leading to stasis, and stasis toxins damaging healthy Qi". The primary therapeutic approach is reinforcing healthy Qi, resolving stasis, and removing toxins, which can reinforce and tonify healthy Qi to regulate pathways, such as phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), C-X-C chemokine receptor type 4/ C-X-C chemokine ligand 12 (CXCR4/CXCL12), and toll-like receptor 4/ nuclear factor-kappa B/ signal transducer and activator of transcription 3 (TLR4/NF-κB/STAT3), adjust T lymphocyte ratios, inhibit Tregs/TAM-M2 function, and downregulate immune checkpoints, including programmed death ligand 1/programmed death 1(PD-L1/PD-1), and reshape TIME. It is also involved in resolving stasis and removing toxins to modulate phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) and hypoxia-inducible factor-1α (HIF-1α) signaling pathways, suppress key enzymes in glycolysis and lipid synthesis, and block toxic metabolite production. Thus, this therapy synergistically regulates the immune and metabolic network, breaks the vicious cycle of "deficiency in healthy Qi and stasis toxins", and offers a novel strategy for integrating traditional Chinese medicine and Western medicine in HCC treatment.
9.HerbGL: a network propagation and graph regularization-based framework for herb pairs prediction in traditional Chinese medicine
Weixiang Liu ; Qian Yuan ; Junjie Zhang ; Xinliang Sun ; Kongfa Hu ; Tao Yang
Digital Chinese Medicine 2026;9(2):265-277
Objective:
To address the challenges of systematically identifying herb pairs in traditional Chinese medicine (TCM), we proposed HerbGL, a framework for predicting potential herb pairs that integrates network propagation and graph regularization.
Methods:
Based on the assumption that herbal actions induce subtle perturbations in biological systems, a framework named HerbGL was proposed. Random walk with restart (RWR) was first applied to the protein-protein interaction (PPI) network to reconstruct herb-specific perturbation effects and generate weighted subnetworks. Then, to quantify affinity between herb pairs, two network-proximity metrics, Closeness and PageRank, were computed from the weighted subnetworks to construct herb-pair affinity matrices. Finally, these matrices, together with known herb pairs (derived from co-occurrence analysis of TCM formulas with a threshold determined from the inflection point of the frequency distribution), were incorporated into a graph regularization model to predict potential herb pairs. Model performance was assessed through baseline comparison, ablation and robustness experiment under different ratios of positive and negative samples, using the area under the receiver operating characteristic curve (AUROC), the area under the precision-recall curve (AUPRC), accuracy, and precision as evaluation metrics. Furthermore, the predicted herb pairs were validated through both literature evidence and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses.
Results:
The weighted subnetworks constructed by RWR provided a refined simulation of herb-specific perturbation effects, which formed the basis for subsequent affinity modeling and prediction. Analysis of herb pair co-occurrence frequencies revealed a marked change around 150, which was selected as the threshold to distinguish herb pairs from non-herb pairs. HerbGL exhibited superior predictive performance compared with baseline models (AUROC = 0.970 5, AUPRC = 0.955 5, accuracy = 0.726 6, precision = 0.970 6). Ablation results showed that removing the Closeness and PageRank metrics substantially degraded performance (AUROC = 0.819 1, AUPRC = 0.876 8), confirming their necessity. Robustness evaluation under an imbalanced positive-to-negative sample ratio of 1 : 5 yielded AUROC = 0.969 6 and AUPRC = 0.840 4, indicating stable predictive ability. Moreover, multiple case studies further validated the rationality of the predicted herb pairs, such as Fangfeng (Saposhnikoviae Radix) and Qingpi (Citri Reticulatae Pericarpium Viride) which are recorded in Liangpeng Huiji (《良朋汇集》, Collection of Excellent Recipes) Vol. 3: Fangfeng Shengma Tang (防风升麻汤). Additionally, pathway enrichment analysis of the Renshen (Ginseng Radix et Rhizoma) and Lianqiao (Forsythiae Fructus) pair further supported the biological plausibility of their compatibility.
Conclusion
HerbGL offers an effective and biologically informed framework for identifying herb pairs in TCM. Beyond improving herb pair prediction, the framework also provides data support for research on herb compounds and mechanisms, thereby supporting data-driven exploration of TCM compatibility.
10.The Potential Role of β-Asarone inCalcium Imbalance and Mitochondrial Dysfunction in Melanoma Cells
Yuze LIU ; Wei TANG ; Biao YU ; Qinghua YANG ; Wenbing LAI
Annals of Dermatology 2026;38(1):75-83
Background:
The treatment landscape for melanoma, a particularly malignant skin cancer, is constrained by notable drug resistance and toxicity. β-Asarone, a natural compound from Acorus tatarinowii, has shown anticancer potential. Disruption of calcium homeostasis and mitochondrial dysfunction are key regulators of tumor cell survival and death.
Objective:
This research was conducted to investigate the impact of β-Asarone on B16F10 melanoma cells, focusing on its potential to induce apoptosis by modulating calcium signaling and mitochondrial function.
Methods:
Cell proliferation and apoptosis were evaluated using CCK-8, colony formation, EdU, and TUNEL assays. Intracellular calcium levels and mitochondrial membrane potential were measured using Fluo-4 AM, Rhod-2 AM, and JC-1 staining. Reactive oxygen species (ROS) generation and adenosine triphosphate (ATP) levels were assessed by fluorescent probes and ATP assay. Western blotting was utilized to detect apoptosis-related proteins, AMP-activated protein kinase (AMPK) pathway activation, and mitochondrial dynamics (OPA1, DRP1, FIS1).
Results:
Treatment with β-Asarone notably inhibited the proliferation of B16F10 cells while simultaneously inducing apoptosis. Fluorescent probe analysis revealed that β-Asarone triggered cytosolic and mitochondrial Ca 2+ overloaded in both the cytosol and mitochondria, accompanied by decreased mitochondrial membrane potential, elevated ROS levels, and reduced ATP production. Western blot analysis showed increased expression of DRP1 and FIS1, decreased OPA1, and enhanced AMPK phosphorylation, indicating that β-Asarone promotes mitochondrial fission through AMPK activation, likely driven by intracellular calcium imbalance.
Conclusion
This study demonstrates that β-Asarone induces apoptosis in B16F10 melanoma cells by triggering Ca 2+ overload and mitochondrial dysfunction.

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