Abstract Background: During prediabetes, increased free fatty acid levels induce negative actions on cardiomyocytes, but effective approaches to prevent these negative actions and the development of diabetic cardiomyopathy are limited. Saturated fatty acids such as palmitate have been shown to contribute to the development of diabetic cardiomyopathy (DbCM), with ferroptosis being recognized as a potential mechanism of palmitate-induced cardiac injury. ALOX15 is a driving factor of ferroptosis and also contributes to inflammation and oxidative stress in DbCM. Baicalein is a natural inhibitor of ALOX15, but the effects of baicalein on ferroptosis in DbCM remain unknown. Aims and Objectives: The overall objective of the present study was to elucidate the potential effects of baicalein on ferroptosis in cardiomyocytes when exposed to elevated free fatty acid levels. Materials and Methods: H9c2 cardiomyocytes were treated with palmitic acid at different concentrations to induce lipid peroxidation and ferroptosis-related responses. The cells were subsequently incubated with baicalein to evaluate its protective effects. Lipid peroxidation and intracellular reactive oxygen species (ROS) levels were measured. Production of 12- and 15-hydroxyeicosatetraenoic acid (15-HETE), as well as mitochondrial function indicators including mitochondrial membrane potential, cytochrome c release, and mitochondrial ROS, were assessed. Protein expression levels of GPX4, ACSL4, and ALOX15 were analyzed by Western blot. Statistical analysis was performed to determine concentration-dependent effects and treatment efficacy. Results: Our results revealed that 12-, 15-HETE production, lipid peroxidation and intracellular reactive oxygen species (ROS) levels increased in a concentration-dependent manner when treated with palmitic acid. Baicalein effectively inhibited the ALOX15 associated 12-, 15- HETEs production and decreased lipid peroxidation in H9c2 cells. Furthermore, baicalein also ameliorated mitochondrial dysfunction through restoring mitochondrial membrane potential and decreasing the release of cytochrome c, as well as mitochondrial ROS. Treatment with baicalein also increased GPX4 and reduced ACSL4 and ALOX15 protein expression in H9c2 cells. Conclusions: These results suggest that baicalein may protect cardiomyocytes against ferroptosis through an ACSL4-ALOX15-GPX4 axis.

Cerebral Small Vessel Disease (CSVD) is a dynamic whole-brain disease, characterized by pathological cascades that affect the brain's venules, capillaries, small arteries, and arterioles. Neuroimaging features of CSVD typically comprise Recent Small Subcortical Infarcts (RSSI), lacunes of presumed vascular origin, White Matter Hyperintensities (WMH) of presumed vascular origin, enlarged Perivascular Spaces (PVS), Cerebral Microbleeds (CMB), and Brain Atrophy (BA). The main clinical features of CSVD often include stroke, abnormal gait, psychiatric disorders, cognitive decline, and urinary incontinence, imposing a heavy burden on individuals and society. Despite its impact, the pathogenesis of CSVD remains unclear, and current clinical diagnosis relies primarily on neuroimaging, presenting considerable challenges for effective treatment. In recent years, most studies have addressed the pathophysiological and molecular mechanisms of CSVD, including chronic cerebral hypoperfusion, inflammatory cascades, oxidative stress, endothelial dysfunction, and Blood-Brain Barrier (BBB) leakage. In addition, genetic factors have been strongly associated with CSVD, though genetic heterogeneity and the complexity of internal environment homeostasis contribute to the persistent uncertainty surrounding its exact mechanisms. A comprehensive overview of these individual mechanisms is crucial for a holistic understanding of the pathogenesis of CSVD. Currently, there is a relative lack of therapeutic drugs and interventions for the complex pathogenesis of CSVD. The existing treatments, such as antihypertensives, antiplatelet agents, lipid-lowering drugs, and hypoglycemic agents, along with traditional alternative therapies like Chinese herbal medicine and acupuncture, have demonstrated efficacy in modulating the occurrence and progression of CSVD. These therapies provide a new perspective for developing more rational CSVD prevention strategies and treatment plans. This review systematically summarizes the cutting-edge research achievements in the field of pathological and physiological mechanisms of CSVD over the past few years, as well as potential treatment pathways and limitations, to provide a theoretical basis and intervention directions for the diagnosis and treatment of this patient population.

BackgroundPanax notoginseng (Burk.) F. H. Chen flower (SQH), a common traditional Chinese medicine and edible food, has long been shown to protect against diabetes. However, the protective effects of SQH against diabetic cardiomyopathy (DbCM) and its potential mechanisms are not yet understood.Aim of the StudyThis study aimed to analyze the therapeutic effects of the flowers of Panax notoginseng (SQH) on DbCM and elucidate its molecular mechanisms.Materials and MethodsThe in vitro model of DbCM was established using palmitate-treated H9c2 cardiomyocyte. A high-fat diet (HFD) in conjunction with streptozotocin (STZ)-induced DbCM mouse model was utilized to validate the cardioprotective effects and mechanism of SQH. Transcriptomic analysis was used to examine the potential mechanisms, followed by both in vitro and in vivo validation of key protein expression levels in the ACSL4/ALOX15 signaling pathway.ResultsIn vitro, SQH treatment significantly protected H9c2 cells from palmitic acid (PA)-induced injury by reducing lipid peroxidation and improving mitochondrial membrane function. Transcriptomic data indicated that SQH influenced ferroptosis-related pathways. Moreover, SQH suppressed the ACSL4/ALOX15 pathway, leading to decreased levels of the key lipid peroxidation product 12-HETE and upregulation of GPX4. In DbCM mice, SQH administration improved glucose homeostasis, attenuated cardiac dysfunction, and reduced myocardial lipid peroxidation.ConclusionThis study demonstrated that SQH ameliorated DbCM injury primarily by inhibiting ACSL4/ALOX15-mediated ferroptosis. These findings not only highlight Panax notoginseng flower as a promising therapeutic candidate for the early intervention of DbCM but also reveal the underlying mechanism of SQH’s protective effect.

BackgroundGranulomatous lobular mastitis (GLM) is a type of non-puerperal mastitis (NPM) associated with autoimmune factors. Previous studies suggest that gut microbiota dysbiosis may play a role in the pathological process of GLM; however, the specific role of gut microbiota metabolites in this process remains unclear. Therefore, this study aimed to investigate the gut metabolic characteristics of short-chain fatty acid (SCFA) in patients with GLM, a type of NPM.MethodsStool samples were collected from 35 patients with GLM and 26 healthy control (HC) subjects. These samples underwent targeted metabolomic analysis to measure SCFA and 16s rRNA high-throughput sequencing to assess gut microbiota composition and differences between the groups.ResultsGas chromatography-mass spectrometry(GC-MS) analysis revealed that the content of SCFA-butanoic acid in the feces of patients with GLM was higher than that in the HC group. Notably, significant differences in metabolic pathways were observed between the HC and GLM groups. High-throughput sequencing results showed that the richness and diversity of gut microbiota in patients with GLM were significantly lower than those in healthy individuals. In addition, 53 bacterial species were found to differ significantly in abundance between the two groups. Moreover, the level of isohexanoic acid in the feces of patients with GLM with recurrence disease was significantly higher than that of patients without recurrence.ConclusionsPatients with GLM exhibit disturbances in gut butanoic acid metabolism and significant differences in gut microbiota structure compared to healthy individuals.

PANoptosis is a novelly defined mode of programmed cell death that involves the activation of multiple cellular death pathways, including pyroptosis, apoptosis, and necroptosis, triggering robust inflammatory reactions. Autophagy is a crucial cellular process that maintains cellular homeostasis and protects cells from various stresses. PANoptosis and autophagy, both vital players in the intricate pathological progression of ischemic stroke (IS), a brain ailment governed by intricate cell death cascades, have garnered attention in recent years for their potential interplay. While mounting evidence hints at a crosstalk between these two processes in IS, the underlying mechanisms remain elusive. Therefore, this review delves into and dissects the intricate mechanisms that underpin the intersection of PANoptosis and autophagy in this devastating condition. In conclusion, the crosstalk between PANoptosis and autophagy in IS presents a promising target for the development of novel stroke therapies. Understanding the interplay between these two pathways offers a much-needed insight into the underlying mechanisms of IS and opens the possibility for new therapeutic strategies.

Microglia/macrophages accumulate at the lesion site by switching toward pro-inflammatory (M1)-dominant phenotype at the acute phase following spinal cord injury (SCI). Such biased polarization shapes the functional outcomes by expanding tissue damage. In the present study, the astrocytic endothelin-1 (ET-1) system is revealed to be immediately activated after SCI, driving microglia polarization toward M1, but suppressing toward M2 phenotype through activation of transcription coactivator YAP via ETA and ETB receptors. In addition, the activation of astrocytic ET-1 system results in elevation of blood plasma ET-1 level, suggesting a high diagnostic value. SCI-induced thrombin is pinpointed as a crucial activator of the astrocytic ET-1 system. The serine protease dramatically promotes the astrocytic expression of preproendothelin-1 (ppET-1) through protease-activated receptor-1 (PAR-1)/RhoA/NF-κB and PAR-1/MAPKs/NF-κB signal pathways. Meanwhile, it induces the expression of astrocytic endothelin-converting enzyme 1 (ECE-1) responsible for mature ET-1 processing. Pharmacological inhibitors of PAR-1 and ET-1 are shown to be highly efficient in microglia M1 phenotype reversion and favorable for the recovery of rat locomotor function after SCI. The findings have revealed a novel mechanism of M1 microglia/macrophages swarming at lesion sites at the acute phase following SCI, and provide potential therapeutic approaches for neuroinflammation by targeting the astrocytic ET-1 system.

Indisulam, a sulfonamide-based compound, is employed as a second-line therapy for NSCLC due to its anti-tumor activity. However, its clinical efficacy is hindered by acquired resistance, the molecular basis of which remains poorly understood. Here, we demonstrate that hypermethylation of RNA-binding protein 39 (RBM39), a specific target of Indisulam, is closely associated with Indisulam resistance. PRMT6 methylates RBM39 at R92. This methylation inhibits Indisulam-induced ubiquitination and proteasomal degradation of RBM39, increases RBM39 protein levels, promotes alternative splicing and expression of proto-oncogenes, and ultimately leads to malignant proliferation and metastasis of NSCLC cells and tumor growth in xenograft mouse models. Inhibiting PRMT6 with MS023 or mutating the RBM39 methylation site enhances Indisulam sensitivity in NSCLC and significantly improves its anti-tumor efficacy. Our findings identify methylated RBM39 as a key biomarker of Indisulam resistance and suggest a potential therapeutic strategy for NSCLC.

Objective To examine the relationship between metabolic syndrome (MS) and its key components in granulomatous mastitis (GM), we explored potential pathological mechanisms through the lens of traditional Chinese medicine (TCM), particularly the concept of intermingled phlegm-blood stasis. Methods In this retrospective study, we enrolled 172 patients with GM and 164 patients with non-inflammatory benign breast masses. Metabolic indicators (waist circumference [WC], blood lipids, etc.), inflammatory markers (C-reactive protein, interleukin-6, tumor necrosis factor-α), and adipose tissue CD68 expression were measured. Logistic regression was used to analyze risk factors and receiver operating characteristic curves were used to evaluate diagnostic efficacy. The correlation between TCM pathogenesis and biomarkers was also examined. Results MS prevalence was significantly higher in the GM group than in the controls (26.16% vs. 6.10%, P < .001). Multivariate analysis identified abdominal obesity (WC ≥ 80 cm, odds ratio [OR] = 1.065) and low levels of high-density lipoprotein cholesterol (HDL-C; <1.29 mmol/L, OR = 0.066) as independent risk factors for GM (P < .001 for both). Among patients with GM, HDL-C levels were inversely correlated with inflammatory markers (r = −0.341 to −0.440), whereas patients with concurrent MS demonstrated greater CD68 macrophage infiltration (P < .001). According to TCM, abdominal obesity corresponds to “spleen deficiency with phlegm-dampness accumulation,” and low HDL-C reflects “deficiency of vital qi,” which collectively lead to phlegm-blood stasis obstruction in the mammary collaterals; this aligns with the key MS driving mechanisms of chronic inflammation and immune dysregulation. Conclusion MS promotes GM development through chronic inflammation and immune dysregulation, with abdominal obesity and low HDL-C levels serving as core risk factors. The TCM theory of intermingled phlegm-blood stasis provides a novel interpretation of the metabolic-inflammatory mechanisms underlying GM. Accordingly, we propose phlegm-resolving and blood-activating strategies as potential therapeutic approaches for metabolic–immune axis regulation.

Background:Convalescent coronavirus disease 2019 (COVID-19) refers to a series of clinical syndromes in patients with COVID-19 infection that follow the relevant discharge indications but do not fulfill the criteria for a clinical cure, and these patients are discharged from the hospital with residual multifunctional deficits, including coughing, fatigue, and insomnia. Due to the prolonged convalescent COVID-19 infection, patients continue to experience symptoms or develop new symptoms after three months of infection, and some symptoms persist for over two months without any apparent triggers, which has a significant impact on the health status and quality of life of the population. Patients with convalescent COVID-19 lack a definitive pharmacological treatment. Traditional Chinese medicine (TCM) exhibits a distinct, synergistic effect on the treatment of convalescent COVID-19. However, there exists a limited number of clinical trials on TCM with lower evidence levels in convalescent COVID-19. Therefore, randomized trials are urgently required. Methods:A multicenter, randomized, double-blind, placebo-controlled, phase II clinical trial was performed to evaluate the efficacy and safety of Shenlingkangfu (SLKF) granules in treating patients with convalescent COVID-19 and lung-spleen qi deficiency syndrome. Eligible participants were aged 18-75 years, had a confirmed severe acute respiratory syndrome coronavirus 2 infection following a positive result for COVID-19 via polymerase chain reaction (PCR) or rapid antigen test at least six months prior, and satisfied clinical criteria. Individuals with a history of severe pulmonary dysfunction or major liver and kidney illness or those on medications were excluded. Multicenter subjects satisfying all criteria were assigned (1:1) randomly into an intervention group and a control group. After a 2-day adjustment period, a total of 154 participants were randomly divided into an intervention group and a control group. The intervention group was given the SLKF granules orally once a bag, 16.9 g, twice daily, whereas the control group received the SLKF granule simulation at the same dosage. The trial was conducted over 1 month, with assessments performed at baseline and 3 months. Results:The primary outcomes were the therapeutic efficacy rate and total clinical symptom score. The secondary outcomes included the fatigue self-assessment scale, pain visual analog scale, Pittsburgh sleep quality index, mini-mental state examination, hospital anxiety and depression scale, TCM syndrome score, C-reactive protein, erythrocyte sedimentation rate, and interleukin-6. Three routine examinations, liver and kidney function tests, and electrocardiography were used as safety indicators. Conclusions:This study aimed to verify whether SLKF granules can significantly improve clinical symptoms, including fatigue, loss of appetite, cough, phlegm, and insomnia, in patients with convalescent COVID-19. For a comprehensive investigation, additional clinical trials with larger sample sizes and longer intervention periods are required.Clinical Trial Registration Center NCT1900024524, Registered on 26 January 2024.

In recent years, the excessive presence of tetracycline antibiotics (TCs) in meat has emerged as a serious threat to human health. In this study, a sensing probe using rare earth europium self-assembled metal-organic skeleton (AlMOF-NH2@Eu) was synthesized for the specific detection of OTC in meat. The -NH2 introduced by the sensing probe can react with OTC through hydrogen bonding, thus greatly quenching the blue fluorescence of AlMOF through IFE, while Eu3+ can effectively enhance the red fluorescence by binding to OTC through antenna effect (AE) with the help of sodium citrate. Meanwhile, the sensing probe demonstrated high selectivity and sensitivity in detecting TCs, and the limits of detection for oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), and doxycycline (DOC) were 0.0128 μM, 0.038 μM, 0.048 μM, and 0.275 μM, respectively. The spiked recovery of meat samples was satisfactory. And the smartphone app-assisted processing of RGB values from the filter paper enabled visual detection of OTC in meat under UV light. These findings highlight the potential applications of AlMOF-NH2@Eu in food for the detection of TCs.

Stress granules (SGs) are considered to be the nonmembrane discrete assemblies present in the cytoplasm to cope with various environmental stress. SGs can promote the progression and drug resistance of hepatocellular carcinoma (HCC). Therefore, it is important to explore the mechanism of SG formation to reduce drug resistance in HCC. In this study, we demonstrate that p110α is required for SGs assembly. Mechanistically, the Arg-Gly (RG) motif of p110α is required for SG competence and regulates the recruitment of SG components. The methylation of p110α mediated by protein arginine methyltransferase 1 (PRMT1) interferes with the recruitment of p110α to SG components, thereby inhibiting the promotion of p110α to SGs. On this basis, we generated metal-polyphenol-network-coated R612F nanoparticles (MPN-R612F), which can efficiently enter HCC cells and maintain the hypermethylation state of p110α, thereby inhibiting the assembly of SGs and ultimately reducing the resistance of HCC cells to sorafenib. The combination of MPN-R612F nanoparticles and sorafenib can kill HCC cells more effectively and play a stronger anti-tumor effect. This study provides a new perspective for targeting SGs in the treatment of HCC.

OBJECTIVES:Astragaloside IV (AST IV) and ligustrazine (Lig), the main ingredients of Astragali Radix and Chuanxiong Rhizoma respectively, have demonstrated significant benefits in treatment of cerebral ischemia -reperfusion injury (CIRI); however, the mechanisms underlying its benificial effects remain unclear. SUMO-1ylation and deSUMO-2/3ylation of dynamin-related protein 1 (Drp1) results in mitochondrial homeostasis imbalance following CIRI, which subsequently aggravates cell damage. This study investigates the mechanisms by which AST IV combined with Lig protects against CIRI, focusing on the involvement of SUMOylation in mitochondrial dynamics.METHODS:Rats were administrated AST IV and Lig for 7 days, and middle cerebral artery occlusion was established to mimic CIRI. Neural function, cerebral infarction volume, cerebral blood flow, cognitive function, cortical pathological lesions, and mitochondrial morphology were measured. SH-SY5Y cells were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) injury. Mitochondrial membrane potential and lactic dehydrogenase (LDH), reactive oxygen species (ROS), and adenosine triphosphate (ATP) levels were assessed with commercial kits. Moreover, co-immunoprecipitation (Co-IP) was used to detect the binding of SUMO1 and SUMO2/3 to Drp1. The protein expressions of Drp1, Fis1, MFF, OPA1, Mfn1, Mfn2, SUMO1, SUMO2/3, SENP1, SENP2, SENP3, SENP5, and SENP6 were measured using western blot.RESULTS:In rats with CIRI, AST IV and Lig improved neurological and cognitive functions, restored CBF, reduced brain infarct volume, and alleviated cortical neuron and mitochondrial damage. Moreover, in SH-SY5Y cells, the combination of AST IV and Lig enhanced cellular viability, decreased release of LDH and ROS, increased ATP content, and improved mitochondrial membrane potential. Furthermore, AST IV combined with Lig reduced the binding of Drp1 with SUMO1, increased the binding of Drp1 with SUMO2/3, suppressed the expressions of Drp1, Fis1, MFF, and SENP3, and increased the expressions of OPA1, Mfn1, Mfn2, SENP1, SENP2, and SENP5. SUMO1 overexpression promoted mitochondrial fission and inhibited mitochondrial fusion, whereas SUMO2/3 overexpression suppressed mitochondrial fission. AST IV combined with Lig could reverse the effects of SUMO1 overexpression while enhancing those of SUMO2/3 overexpression.CONCLUSIONS:This study posits that the combination of AST IV and Lig has the potential to reduce the SUMO-1ylation of Drp1, augment the SUMO-2/3ylation of Drp1, and thereby exert a protective effect against CIRI.

BackgroundCerebral ischemia/reperfusion injury (CIRI) is a sequence of pathophysiological processes after blood recanalization in the patients with ischemic stroke, and has become the hinder for the rehabilitation. Naotaifang formula (NTF) has exhibited the clinical effectiveness for this disease. However, its action effects and molecular mechanisms against CIRI are not fully elucidated.PurposeThe research was to clarify the crosstalk between ferroptosis and necroptosis in CIRI, and uncover the mechanism underlying the neuroprotection of NTF.MethodsThis study established MCAO/R rat models with various reperfusion times. Western blot, transmission electron microscope, laser speckle imaging, immunofluorescence, immunohistochemistry and pathological staining were conducted to detect and analyze the obtained results. Subsequently, various NTF doses were used to intervene in MCAO/R rats, and biology experiments, such as western blot, Evans blue, immunofluorescence and immunohistochemistry, were used to analyze the efficacy of NTF doses. The effect of NTF was further clarified through in vitro experiments. Eventually, HT22 cells that suffered OGD/R were subjected to pre-treatment with plasmids overexpressing HSP90, MLKL, and GPX4 to indicate the interaction among ferroptosis and necroptosis.ResultsThere was a gradual increase in the Zea Longa score and cerebral infarction volume following CIRI with prolonged reperfusion. Furthermore, the expression of factors associated with pro-ferroptosis and pro-necroptosis was upregulated in the cortex and hippocampus. NTF alleviated ferroptosis and necroptosis in a dose-dependent manner, downregulated HSP90 levels, reduced blood-brain barrier permeability, and thus protected nerve cells from CIRI. The results in vitro research aligned with those of the in vivo research. HSP90 and MLKL overexpression promoted necroptosis and ferroptosis while activating the GCN2-ATF4 pathway. GPX4 overexpression had no effect on necroptosis or the associated signaling pathway. The administration of NTF alone, as well as its combination with the overexpression of HSP90, MLKL, or GPX4 plasmids, decreased the expression levels of factors associated with pro-ferroptosis and pro-necroptosis and reduced the protein levels of the HSP90-GCN2-ATF4 pathway. Moreover, the regulatory effects of the NTF alone group on GSH, ferrous iron, and GCN2 were more significant compared with those of the HSP90 overexpression combination group.ConclusionFerroptosis and necroptosis were gradually aggravated following CIRI with prolonged reperfusion. MLKL overexpression may promote ferroptosis and necroptosis, while GPX4 overexpression may have little effect on necroptosis. HSP90 overexpression accelerated both forms of cell death via the HSP90-GCN2-ATF4 pathway. NTF alleviated ferroptosis and necroptosis to attenuate CIRI by regulating the HSP90-GCN2-ATF4 pathway. Our research provided evidence for the potential of drug development by targeting HSP90, MLKL, and GPX4 to protect against ischemic stroke.

Ethnopharmacological relevance: Cerebral ischemia-reperfusion injury (CIRI) is a complex pathophysiological process involving multiple factors, and becomes the footstone of rehabilitation after ischemic stroke. Sanpian decoction (SPD) has exhibited protective effects against CIRI, migraine, and other cerebral vascular diseases. However, the underlying mechanisms have not been completely elucidated. Aim of the study: This study sought to explore the potential mechanisms underlying the effect of SPD against CIRI. Materials and methods: High-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UPLC) were carried out to determine the chemical constituents of SPD. A network pharma-cology approach combined with experimental verification was conducted to elucidate SPD's multi-component, multi-target, and multi-pathway mechanisms in CIRI occurrence. The pharmacodynamics of the decoction was evaluated by establishing the rat model of middle cerebral artery occlusion/reperfusion (MCAO/R). In vivo and in vitro experiments were carried out, and the therapeutic effects of SPD were performed using 2,3,5-triphenyltetra-zolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, and Nissl staining. We used terminal deoxy-nucleotidyl transferase dUTP nick end labeling (TUNEL) staining and flow cytometry to evaluate cortex apoptosis. The quantification of mRNA and corresponding proteins were performed using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot respectively. Results: Our research showed that pretreatment with SPD improved neurological function and inhibited CIRI. Network pharmacology revealed that the hypoxia-inducible factor-1 (HIF-1) signaling pathway and mitogen-activated protein kinase (MAPK) signaling pathway-mediated apoptosis may be associated with CIRI. In vivo and in vitro experiments, we confirmed that SPD increased cerebral blood flow, improved neural function, and reduced neural apoptosis via up-regulating the expression of sirtuin 1 (SIRT1) and down-regulating phospho-extracellular regulated protein kinases (p-ERK)/ERK and HIF-1 & alpha; levels in CIRI rats. Conclusion: Taken together, the present study systematically revealed the potential targets and signaling path-ways of SPD in the treatment of CIRI using in silico prediction and verified the therapeutic effects of SPD against CIRI via ameliorating apoptosis by regulating SIRT1/ERK/HIF-1 & alpha;.

AimTo explore the molecular mechanism of Sijunzi Decoction (SJZD) in the treatment of Parkinson’s disease (PD) through the application of network pharmacology, molecular docking, and molecular dynamics simulations, complemented by experimental verification.MethodsThe BATMAN-TCM, GeneCards, and DisGeNet databases were searched to screen the active components and therapeutic targets of SJZD. Cytoscape (3.7.1) was used to create a network diagram of the components and targets. The STRING platform was used to construct a protein-protein interaction (PPI) network. The Bioconductor database and RX64 (4.0.0) software were used to conduct Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis on the core target genes. The binding sites and binding energies between SJZD active components and the target were analyzed by molecular docking and dynamic simulation. Finally, the therapeutic effect and mechanism of SJZD were verified by Cell Counting Kit-8 (CCK-8) and Western blotting (WB).ResultsThis research identified 188 active compounds in SJZD, 1568 drug targets, 2069 PD targets, and 451 intersection targets related to PD. According to network analysis, Adenosine Triphosphate, Tridecanoic Acid, Hexadecanoic Acid, Pentadecanoic Acid, and Adenosine were identified as the core components of SJZD in the treatment of PD. The five targets with the highest Degree values in the PPI network were AKT1, INS, TNF, IL-6, and TP53. The GO and KEGG enrichment analyses, in turn, determined that the administration of SJZD for the treatment of PD may engage processes such as xenobiotic stimulation and biological stimulus response. Furthermore, AGE-RAGE and cAMP signaling pathways related to diabetic complications may be involved. Molecular docking and kinetic simulations showed that IL-6 and AKT1 bind best to Adenosine. Experimental results showed that SJZD significantly reduced 6-OHDA-induced apoptosis of SH⁃SY5Y cells by activating the PI3K/AKT signaling pathway and regulating the expression of apoptosis factors such as Bcl⁃2 and Bax.ConclusionSJZD is essential in the processes of apoptosis and neuronal protection, acting through various components that target multiple pathways. Notably, the PI3K/AKT pathway is a verified SJZD-PD target, providing a reference for clinical precision drug use for PD.

We aimed to explore the molecular mechanism of Ruxian Shuhou prescription in the treatment of triple-negative breast cancer (TNBC) by using network pharmacology. The active components and targets of the prescription were obtained by Traditional Chinese medicine systems pharmacology database. Gencards database, online mendelian inheritance in man database, therapeutic target database, and DRUGBANK database were used to search for the TNBC-related targets. The potential targets of Ruxian Shuhou prescription for TNBC were screened out by the intersection of effective ingredient action targets and disease targets. A herb-active ingredient-target network was constructed and analyzed for key ingredients. A protein-protein interaction network was constructed for studying key targets. Furthermore, gene ontology analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis were carried out. Finally, the relationship between key ingredients and key genes was evaluated by molecular docking. The key ingredients of Ruxian Shuhou prescription for the treatment of TNBC may be Quercetin, Luteolin and Kaempferol, while the key therapeutic targets may be protein kinase B, interleukin-6, cellular tumor antigen p53, and vascular endothelial growth factor A. The related signaling pathways were mainly involved in tumor, apoptosis and virus infection, among which the PI3K-Akt signaling pathway was the most closely related to TNBC. Molecular docking showed that the key ingredients had high binding activity with the key targets. The molecular mechanisms of Ruxian Shuhou prescription for TNBC are likely to involve multi-ingredient, multi-target and multi-pathway.

Objective To analyze the current status and development trends of the patents of spleen-in-vigorating health food with the homology of medicine and food in China,and to provide ideas and references for the research and development of traditional Chinese medicine(TCM)spleen-invigorating health food with the homology of medicine and food. Methods The State Administration for Market Regulation website's"Special Food Informa-tion Query Platform"and the incoPat global patent database were searched in this study.Based on the methods of bibliometrics,the registered health food and patents related to spleen-invigorating health food with the homology of medicine and food in China were sort-ed out.Furthermore,the research and development numbers,provinces,institutions,tech-nology and efficacy classification,major drugs,active ingredients and others of invigorating spleen health food in China were analyzed,and filtered patent data were visualized and ana-lyzed by R programming language and CytoScape software. Results A total of 285 patents of health food with the homology of medicine and food for in-vigorating spleen were included and analyzed.From 2012,the patent registration numbers of these spleen-invigorating health food with the homology of medicine and food increased sig-nificantly in China.Over the past 20 years,the top five provinces in terms of patent disclo-sures were Guangdong,Anhui,Jiangsu,Shandong,and Guangxi.It was found that the techni-cal efficacy of over 20 patents was described as"immune enhancement""digestion""disease prevention",etc.Patent applications were mainly aimed at the research and development of the preservation of food or ingredients,the specific therapeutic activity of compounds,and pharmaceutical preparations,which were led by corporation research and development registrations,and supplemented by applications from research institutions and individuals.Among the 285 patents,the top 10 raw materials of spleen-invigorating health food with the homology of medicine and food were Shanyao(Dioscoreae Rhizoma),Fuling(Poria),Shanzha(Crataegi Fructus),Baizhu(Atractylodis Macrocephalae Rhizoma),Chenpi(Citri Reticulatae Pericarpium),Dazao(Jujubae Fructus),Gancao(Glycyrrhizae Radix et Rhizoma),Fengmi(Mel),Maiya(Hordei Fructus Germinatus),and Dangshen(Salviae Miltiorrhizae Radix et Rhizoma).The main functions were to nourish spleen and replenish Qi,invigorate spleen and benefit lungs,nourish blood and promote fluid production,and nourish spleen and stomach. Conclusion The main drug composition and functional components of spleen-invigorating health food with the homology of medicine and food are relatively clear,and the technical ef-fects of invigorating the spleen and stomach,eliminating accumulation of food,and enhanc-ing immunity are highly targeted.This paper provides evidence for the research and develop-ment,mechanism research,and process improvement of spleen-invigorating health food with the homology of medicine and food in the future.

Cerebral ischemia, a leading cause of disability and mortality worldwide, triggers a cascade of molecular and cellular pathologies linked to several central nervous system (CNS) disorders. These disorders primarily encompass ischemic stroke, Alzheimer's disease (AD), Parkinson's disease (PD), epilepsy, and other CNS conditions. Despite substantial progress in understanding and treating the underlying pathological processes in various neurological diseases, there is still a notable absence of effective therapeutic approaches aimed specifically at mitigating the damage caused by these illnesses. Remarkably, ischemia causes severe damage to cells in ischemia-associated CNS diseases. Cerebral ischemia initiates oxygen and glucose deprivation, which subsequently promotes mitochondrial dysfunction, including mitochondrial permeability transition pore (MPTP) opening, mitophagy dysfunction, and excessive mitochondrial fission, triggering various forms of cell death such as autophagy, apoptosis, as well as ferroptosis. Ferroptosis, a novel type of regulated cell death (RCD), is characterized by iron-dependent accumulation of lethal reactive oxygen species (ROS) and lipid peroxidation. Mitochondrial dysfunction and ferroptosis both play critical roles in the pathogenic progression of ischemia-associated CNS diseases. In recent years, growing evidence has indicated that mitochondrial dysfunction interplays with ferroptosis to aggravate cerebral ischemia injury. However, the potential connections between mitochondrial dysfunction and ferroptosis in cerebral ischemia have not yet been clarified. Thus, we analyzed the underlying mechanism between mitochondrial dysfunction and ferroptosis in ischemia-associated CNS diseases. We also discovered that GSH depletion and GPX4 inactivation cause lipoxygenase activation and calcium influx following cerebral ischemia injury, resulting in MPTP opening and mitochondrial dysfunction. Additionally, dysfunction in mitochondrial electron transport and an imbalanced fusion-to-fission ratio can lead to the accumulation of ROS and iron overload, which further contribute to the occurrence of ferroptosis. This creates a vicious cycle that continuously worsens cerebral ischemia injury. In this study, our focus is on exploring the interplay between mitochondrial dysfunction and ferroptosis, which may offer new insights into potential therapeutic approaches for the treatment of ischemia-associated CNS diseases.

Background: Hypertensive cerebral small vessel disease (HT-CSVD) is a cerebrovascular clinical, imaging and pathological syndrome caused by hypertension (HT). The condition manifests with lesions in various vessels including intracranial small/arterioles, capillaries, and small/venules. Hypertensive cerebral small vessel disease has complex and diverse clinical manifestations. For instance, it can present as an acute stroke which progresses to cause cognitive decline, affective disorder, unstable gait, dysphagia, or abnormal urination. Moreover, hypertensive cerebral small vessel disease causes 25–30% of all cases of ischemic strokes and more than 50% of all cases of single or mixed dementias. The 1-year recurrence rate of stroke in cerebral small vessel disease patients with hypertension is 14%. In the early stage of development, the symptoms of hypertensive cerebral small vessel disease are concealed and often ignored by patients and even clinicians. Patients with an advanced hypertensive cerebral small vessel disease manifest with severe physical and mental dysfunction. Therefore, this condition has a substantial economic burden on affected families and society. Naotaifang (NTF) is potentially effective in improving microcirculation and neurofunction in patients with ischemic stroke. In this regard, this multicenter randomized controlled trial (RCT) aims to furtherly evaluate the efficacy and safety of naotaifang capsules on hypertensive cerebral small vessel disease.Methods: This study is a multicenter, randomized, double-blind, placebo-controlled clinical trial. A total of 388 eligible subjects were recruited from the First Hospital of Hunan University of Chinese Medicine, Hunan Academy of Chinese Medicine Affiliated Hospital, the First Hospital of Shaoyang University, the First Traditional Chinese Medicine Hospital of Changde, and Jiangmen Wuyi Hospital of Traditional Chinese Medicine from July 2020 to April 2022. After a 4-week run-in period, all participants were divided into the intervention group (represented by Y-T, N-T) and control group (represented by Y-C, N-C); using a stratified block randomized method based on the presence or absence of brain damage symptoms in hypertensive cerebral small vessel disease (represented by Y and N). The Y-T and N-T groups were administered different doses of naotaifang capsules, whereas Y-C and N-C groups received placebo treatment. These four groups received the treatments for 6 months. The primary outcome included Fazekas scores and dilated Virchow-robin spaces (dVRS) grades on magnetic resonance imaging (MRI). The secondary outcomes included the number of lacunar infarctions (LI) and cerebral microbleeds (CMB) on magnetic resonance imaging, clinical blood pressure (BP) level, traditional Chinese medicine (TCM) syndrome scores, mini-mental state examination (MMSE) scale, and safety outcomes. Fazekas scores, dilated Virchow-robin spaces grades, and the number of lacunar infarctions and cerebral microbleeds on magnetic resonance imaging were tested before enrollment and after 6 months of treatment. The clinical blood pressure level, traditional Chinese medicine syndrome scores, mini-mental state examination scale and safety outcomes were tested before enrollment, after 3-month, 6-month treatment and 12th-month follow-up respectively.Conclusion: The protocol will comfirm whether naotaifang capsules reduce Fazekas scores, dilated Virchow-robin spaces grades, and the number of lacunar infarctions and cerebral microbleeds, clinical blood pressure, increase mini-mental state examination scores, traditional Chinese medicine syndrome scores of Qi deficiency and blood stasis (QDBS), and improve the quality of life of subjects. The consolidated evidence from this study will shed light on the benefits of Chinese herbs for hypertensive cerebral small vessel disease, such as nourishing qi, promoting blood circulation and removing blood stasis, and dredging collaterals. However, additional clinical trials with large samples and long intervention periods will be required for in-depth research.Clinical Trial registration:www.chictr.org.cn, identifier ChiCTR1900024524.

The induction of ferroptosis in tumor cells is one of the most important mechanisms by which tumor progression can be inhibited; however, the specific regulatory mechanism underlying ferroptosis remains unclear. In this study, we found that transcription factor HBP1 has a novel function of reducing the antioxidant capacity of tumor cells. We investigated the important role of HBP1 in ferroptosis. HBP1 down-regulates the protein levels of UHRF1 by inhibiting the expression of the UHRF1 gene at the transcriptional level. Reduced levels of UHRF1 have been shown to regulate the ferroptosis-related gene CDO1 by epigenetic mechanisms, thus up-regulating the level of CDO1 and increasing the sensitivity of hepatocellular carcinoma and cervical cancer cells to ferroptosis. On this basis, we constructed metal-polyphenol-network coated HBP1 nanoparticles by combining biological and nanotechnological. MPN-HBP1 nanoparticles entered tumor cells efficiently and innocuously, induced ferroptosis, and inhibited the malignant proliferation of tumors by regulating the HBP1-UHRF1-CDO1 axis. This study provides a new perspective for further research on the regulatory mechanism underlying ferroptosis and its potential role in tumor therapy.