Along with more research into the mechanisms of pulmonary fibrosis, anti-pulmonary fibrosis drugs under development are becoming diversified. Over the past decade, multiple phase Ⅱ/Ⅲ clinical trials have been terminated due to insufficient efficacy, with seven single-target monoclonal antibody drugs failing to meet expectations. In contrast, multi-target drugs such as nintedanib and pirfenidone have been successfully marketed, demonstrating favorable clinical outcomes. However, drugs 
directly targeting transforming growth factor-β have raised safety concerns. Ongoing phase Ⅲ candidates, such as lysophosphatidic acid receptor 1 antagonists and phosphodiesterase 4B inhibitors, do not directly intervene in the transforming growth factor-β signaling pathway. Given the potential limitations of single-target drugs, future drug development is expected to prioritize multi-target and multi-cell strategies while exploring synergistic multi-drug therapies. This article reviews the current clinical trials for anti-pulmonary fibrosis drugs worldwide and the challenges they face in order to provide references for the research and development of new anti-pulmonary fibrosis drugs.

Metabolism-related diseases are chronic diseases caused by genetic and environmental factors. The symptoms include insulin resistance, abnormal blood glucose and lipid levels, and elevated blood pressure. This type of illness has become a major threat to human health, and there is an urgent need to find effective treatments. Advanced glycation end products (AGE) are a group of complex and heterogeneous compounds that result from reduced interactions between the carbonyl groups of  sugar and the free amino groups of proteins, lipids, and nucleic acids. Increasing evidence shows that AGE and its receptor (RAGE) are involved in the occurrence and development of such metabolism-related diseases as hypertension, diabetes, and atherosclerosis. AGE can have adverse effects on tissues through non-receptor and receptor-mediated mechanisms. In the receptor-mediated mechanism, AGE interacts with RAGE to increase the production of oxygen free radicals and activate NF-κB so that more pro-inflammatory cells are expressed and released, leading to cell damage. This article reviews the research progress in interventions with AGE and RAGE in the treatment of hypertension, diabetes, and atherosclerosis from a metabolic perspective in the hope of exploring the potential of AGE and RAGE as therapeutic targets for metabolism-related diseases.

The Hippo/YAP signaling pathway is an evolutionarily conserved protein kinase cascade that plays an important role in a variety of biological processes, such as cell proliferation and differentiation, organ growth and tissue regeneration. Fibrosis is a continuous and highly dynamic process characterized by excessive deposition of extracellular matrix, resulting in irreversible pathological changes that eventually lead to the failure of multiple tissues and organs. Targeted therapeutic strategies to ameliorate or reverse fibrosis are lacking. Studies have shown that the aberrantly activated Hippo/YAP signaling pathway may play a role in the development of fibrosis by regulating collagen deposition, fibroblast overproliferation, and epithelial cell differentiation, but the specific mechanism of action has not been fully elucidated. Targeting the Hippo/YAP signaling pathway involves two mechanisms: one is to target the upstream molecules of the Hippo/YAP signaling pathway, which is mainly achieved by inhibiting the activity of the core kinase or blocking the interaction with other molecules; the other is to target the downstream activities of YAP/TAZ and YAP/TAZ-TEAD in the Hippo/YAP signaling pathway. Studies have shown that the phosphorylation and subcellular localization of YAP/TAZ are significantly altered when tissue and organ damage occurs. This article is intended to review the current research on Hippo/YAP signaling pathway and its mediation of fibrosis in the lung, heart, liver, kidney, pancreas and skin in hopes of providing new ideas for studies on the pathogenesis of fibrosis and targeted therapeutic drugs.

Methamphetamine (METH) is a highly addictive synthetic psychoactive drug that is often abused as a psychostimulant. Chronic exposure to METH induces neurotoxic effects through oxidative stress, impairment of mitochondrial function, activation of astrocytes and microglia, and amino acid excitability. METH abuse increases the chance of developing neurodegenerative diseases such as Alzheimer′s disease, Parkinson′s disease, Huntington′s disease. However, the mechanism behind METH-induced neurotoxicity remains incompletely understood. So far, there is no specific treatment for METH-induced neurotoxicity. This paper reviews some of the potential mechanisms of METH-induced neurotoxicity in recent years, such as neuroinflammation, glutamatergic excitotoxicity, oxidative stress, and mitochondrial toxicity, and discusses the current therapeutic strategies related to mitigating the neurotoxic effects of METH in the brain through different pathways.

Commonly used anesthetic sedatives (opioids, benzodiazepines, ketamine, propofol, etc.) share the risk of inducing respiratory depression, and their multi-target mechanism of action presents significant heterogeneity. Opioids inhibit the rhythmic activity of the respiratory center of the medulla bulbar (such as the PreBötzinger complex and parbrachial nucleus) by activating both the μ-opioid receptor and the G-protein-gated inwardly-rectifying potassium channel and β-arrestin signaling pathway, resulting in decreased respiratory frequency and amplitude. Benzodiazepines enhance inhibitory neurotransmission mediated by γ-aminobutyric acid receptors, reduce the sensitivity of chemoreceptors to PaCO₂ and PaO₂, and lead to a decreased tidal volume and upper airway obstruction. Ketamine inhibits respiratory drive and respiratory muscle function by blocking N-methyl-D-aspartic acid  receptors and indirectly affecting the μ-opioid receptor. In addition, propofol inhibits pre-expiratory neuronal activity and relaxes upper airway muscles by activating the GABA_A receptor β₃ subunit. Currently, specific antagonists (naloxone/flumazenil) and respiratory stimulants (doxapram) are clinically used to treat respiratory depression, but they have defects such as short duration of action and insufficient specificity. The development of novel stimulants targeting μ-opioid receptor agonists and the D-serine release pathway of astrocytes, as well as broad-spectrum antidotes based on "molecular cage" technology, has become a new sphere of research that aims at precisely reversing respiratory depression while preserving analgesic and sedative effects. This article reviews the biological mechanisms of respiratory depression caused by sedative hypnotic anesthetic drugs, explores the advantages and disadvantages of treatments currently availabe, and proposes new strategies for improving respiratory depression in the future.

Nanomaterials have been used in a variety of industries recently, involving foods, chemicals and biomedicine. There are multiple routes through which humans are exposed to nanomaterials, and their toxic effects deserve more attention. In vivo studies have confirmed that nanomaterials exposure can lead to toxicity in such target organs as the heart, liver, kidney, skin and nerve. The toxicity mechanism is related to changes in organelles such as the endoplasmic reticulum, lysosomes and mitochondria. Increasing studies suggest that mitochondria are critical targets for the toxicity of nanomaterials. Mitochondrial biogenesis serves as an important mechanism for maintaining mitochondrial homeostasis, which plays a vital role in the process of nanomaterials-induced cellular toxicity. This article summarizes the current research on the effects of nanomaterials on mitochondrial biogenesis, and elaborates the mechanism through which nanomaterials disrupt mitochondrial biogenesis by triggering oxidative stress, upsetting the homeostasis of calcium ion and disturbing toxicity pathways. This article is expected to provide a reference for toxicity testing and risk assessment of nanomaterials.

Antibody-based therapies are one of the crucial tumor-targeted therapies, enabling precise elimination of tumor cells by specifically binding to antigens on the tumor cell surface. However, their wide applications in solid tumor therapy are often limited by on-target toxicity. Recent advancements in antibody engineering have led to the development of novel tumor-targeted masking antibodies, which are specifically designed to address these limitations. Masking antibodies typically consist of an antibody domain, a masking domain and a linker. These antibodies are characterized by selective activation and other functional properties. Currently, various masking antibody technologies with distinct characteristics have been developed and have demonstrated favorable safety profiles in animal studies. This review summarizes the structure and characteristics of tumor-targeted masking antibodies outlines common masking technologies and their drug development in order to offer new lines of thought  for the design and development of next-generation tumor-targeted therapeutics.

Sleep disorders are becoming a social problem that poses a danger to human health and an obstacle to economic development. As an endogenous hormone, melatonin plays an important role in regulating circadian rhythm. Research has shown that exogenous melatonin holds promising prospects in the treatment of insomnia disorders and thus merits in-depth exploration and development. Given the close relationship between pharmacokinetic properties and drug efficacy as well as toxicity, this paper reviews the quantitative analysis methods for melatonin and its major metabolites, especially the  pretreatment methods for biological samples and the related LC-MS/MS analysis methods. Studies related to the absorption, distribution, metabolism, excretion and drug-drug interactions of melatonin both in vivo and in vitro, as well as the level and species differences of endogenous melatonin are also outlined. Meanwhile, the druggability of melatonin and corresponding solutions are explored in the hope of providing data for the development of exogenous melatonin into novel sleep-regulating drugs.

OBJECTIVE  To investigate the mechanism by which salvianolic acid A (Sal A) reduces the inflammatory response and oxidative stress of BV2 cells injured by oxygen and glucose deprivation/reperfusion (OGD/R). METHODS  An OGD/R injury model of BV2 cells was established with sugar free Earle solution containing Na₂S₂O₄ 10 mmol·L^-1. Na₂S₂O₄ sugar free Earle solution was added and cultured in an incubator (37 ℃, 5%CO₂) for 1.5 h (oxygen glucose deprivation) before a normal medium was used for 24 h (reperfusion). Then, the cells were divided into the cell control group, OGD/R group, OGD/R+Sal A 1, 5 and 10 μmol·L^-1 group, OGD/R+ML385 group, OGD/R+ML385+Sal A 1, 5 and 10 μmol·L^-1 group and OGD/R+edaravone (Eda, 50 μmol·L^-1) group. After twenty-four hours of culture, the cell survival rate was measured by CCK8 kit. The contents of Interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), IL-10, IL-4 and transforming growth factor-β (TGF-β) in the cell supernatant were detected by ELISA. Reactive oxygen species (ROS) in cells was detected using the chemical fluorescence method. The contents of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and chloramphenicol acetyltransferase (CAT) in cells were determined with the colorimetric method. Protein expressions of Kelch like ECH-associated protein 1 (Keap1), nuclear factor erythroid-2 related factor 2 (Nrf2),Heme oxygenase-1 (HO-1), NADPH: quinone oxidoreductase-1 (NQO1) and p-nuclear factor kappa-B p65 (p-NF-κB p65) were detected by Western blotting. RESULTS  ① Compared with the cell control group, the cell survival rate of the OGD/R group was significantly decreased (P<0.01). Compared with the OGD/R group, the survival rates of OGD/R+Sal A 1, 5 and 10 μmol·L-1 groups were significantly increased (P<0.05, P<0.01). ② Compared with the cell control group, the contents of IL-1β, IL-6 and TNF-α were significantly increased, the contents of IL-10, IL-4 and TGF-β were significantly decreased, the contents of ROS and MDA were significantly increased, and the activities of SOD, CAT and GSH-Px were significantly decreased in the OGD/R group (P<0.01). Compared with the OGD/R group, the content of IL-6 was significantly decreased, the contents of IL-10, IL-4 and TGF-β were significantly increased, the contents of ROS and MDA were significantly decreased, and the activities of SOD, CAT and GSH-Px were significantly increased in OGD/R+Sal A 1, 5 and 10 μmol·L-1 and OGD/R+Eda groups (P<0.05, P<0.01). ③ Compared with the cell control group, the protein expression of p-NF-κB P65 in the OGD/R group was significantly increased (P<0.01). Compared with the OGD/R group, the protein expressions of Keap1 and cytoplasmic Nrf2 were significantly decreased, the expressions of nuclear Nrf2, HO-1 and NQO1 proteins were significantly increased, and the expression of p-NF-κB p65 protein was significantly decreased in OGD/R+Sal A 5 and 10 μmol·L^-1 and OGD/R+Eda groups (P<0.05, P<0.01). In OGD/R+ML385, OGD/R+ML385+Sal A 1, 5 and 10 μmol·L^-1 groups, the protein expression of Keap1 was significantly increased, the protein expressions of cytoplasmic Nrf2, nuclear Nrf2, HO-1 and NQO1 protein were significantly decreased, and the protein expression of p-NF-κB P65 was significantly increased (P<0.01). CONCLUSION  Sal A reduces the inflammatory response and oxidative stress of OGD/R injured BV2 cells possibly by activating the Keap1/Nrf2 pathway and inhibiting the NF-κB pathway.

As antibiotic resistance becomes increasingly concerning, antimicrobial peptides, as a new type of antibiotic alternative, have attracted more attention. However, the low enzymatic stability of antimicrobial peptides severely limits their clinical applications. To address this issue, researchers have developed various structural modification strategies, including the introduction of unnatural amino acids, peptide chain cyclization and chemical group modification. This article reviews the basic principles and cases of the above modification strategies analyzes the advantages and disadvantages of different modification strategies and recommends ways these strategies can be optimized. In addition, this article predicts the developments of and potential challenges to strategies for enhancing enzymatic stability of antimicrobial peptides in the hope of providing references for subsequent research and development of antimicrobial peptides.

Special populations such as the elderly, pregnant women, children, and patients with impaired liver and kidney function have different physiological characteristics and drug processes from other patients. As a result during medication, these populations are vulnerable to drug-drug interactions in case of combined medications, which makes it more difficult of optimize drug treatment and develop new drugs. The physiologically based pharmacokinetic (PBPK) model can predict drug-drug interaction in special populations by altering metabolism-related physiological parameters such as metabolic 
enzymes, transporter activity and clearance. Gene polymorphisms, intestinal metabolism and other 
factors also affect the accuracy of model results. This review aims to to optimize clinical regimens for special populations and provide references for new drug development.

OBJECTIVE  To investigate the role of Bai Ling Long Zao An Shen formula (BLLZ) in sleep improvement in an environmental stress-induced insomnia rat model and explore its underlying mechanisms. METHODS  (1) Component analysis: the chemical constituents of the BLLZ extract were analyzed using ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS). (2) Evaluation of the sedative and hypnotic effect: ① Mice: 50 ICR mice were randomly divided into normal control group, BLLZ-L group (5, 10 and 20 g·kg^-1) and diazepam group (DZP, 3 mg·kg^-1). After five days of intragastric administration, pentobarbital sodium-induced righting reflex and locomotor activity tests were performed. ② Rats: 8 SD rats were implanted with electrodes and allowed to recover for seven days before baseline EEG data was collected over 24 h. A crossover design (7 d washout period) was employed,  with rats randomly assigned to the DZP (3 mg·kg^-1) and BLLZ (20 g·kg^-1) group. After five days of treatment, 24 h EEG recordings were obtained. (3) Insomnia model and interventions: ① 8 SD rats were allowed to recover for seven days post-surgery, followed by 6 h (14:00-20:00) baseline EEG recording. A 3×3 crossover design was used to assign rats to model (environmental stress-induced insomnia), model+DZP, or model+BLLZ groups. After five days of treatment, insomnia was induced by frequent cage changes (14:00, 16:00 and 18:00), and EEG changes were monitored. (4) Mechanistic study: 32 SD rats were randomly divided into the normal control group, model group, and model+DZP group. After five days of treatment, hypothalamic tissues were collected for biochemical analysis. γ-aminobutyric acid (GABA), glutamate (Glu), and dopamine (DA) levels were measured 
using biochemical kits while γ aminobutyric acid receptor subunit alpha-1 (GABAA1), core clock proteins period circadian regulator 2 (PER2) and circadian locomotor output cycles (CLOCK) protein expressions were assessed by Western blotting. RESULTS  (1) Compared with the normal control group, the sleep latency of BLLZ 10 and 20 g·kg^-1 and DZP groups was significantly shortened, and the locomotor activity of BLLZ 20 g·kg^-1 and DZP groups was significantly reduced; BLLZ 20 g·kg^-1 significantly increased the total sleep time, slow-wave sleep time, and average duration of sleep in normal rats, and significantly reduced the wakefulness time. (2) The total sleep time and slow-wave sleep time of the model group significantly decreased and the wakefulness time significantly increased compared with baseline. (3) Compared with the model group, the total sleep time and slow-wave sleep time of the model+BLLZ group and the model+DZP group were significantly increased, and the wakefulness time significantly shortened. (4) Compared with the normal control group, the Glu/GABA ratio, DA content and CLOCK protein expression were significantly increased and GABAA1 and PER2 protein expression were significantly decreased in the model group; compared with the model group, the Glu/GABA ratio, DA content and CLOCK protein expression were significantly decreased, and the expression of GABAA1 and PER2 were significantly increased in the model+BLLLZ group and the model+DZP group. CONCLUSION  BLLZ has sedative and hypnotic effects. It can prolong the total slow-wave sleep time by increasing the average duration of slow-wave sleep episodes, thereby increasing the total sleep time and improving environmental stress-induced insomnia. The mechanism may be related to the downregulation of the Glu/GABA ratio and DA levels as well as the enhancement of GABAA1 
expressions and the regulation of hypothalamic core clock protein expressions.

Acral melanoma (AM) is a distinct and aggressive subtype of melanoma characterized by high metastatic potential and poor prognosis. The pathogenesis and therapeutic strategies for advanced AM differ significantly from those of other melanoma subtypes, yet AM has received relatively less attention. AM exhibits high heterogeneity and a low tumor mutation burden. Mutations in braf, nras, and the tert promoter occur at much lower frequencies in AM than in cutaneous melanoma, limiting the 
effectiveness of treatments such as recombinant B-Raf proto oncogene serine/threonine protein kinase(BRAF) inhibitors. Additionally, reduced tumor immunogenicity due to low tumor-infiltrating lymphocyte levels contributes to the limited efficacy of immune checkpoint inhibitors, including anti-programmed death-1 and anti-cytotoxic T-lymphocyte-associated protein 4 therapies. Recent discoveries of novel therapeutic targets, such as receptor tyrosine kinases and cyclin-dependent kinases, along with emerging immune checkpoints, including V-domain immunoglobulin suppressor of T cell activation, adenosine A2A receptor, T cell immunoglobulin and ITIM domain, and T cell immunoglobulin and mucin-domain containing-3, offer new prospects for improving AM patient outcomes. Many AM treatments remain in the experimental stage, with research focusing on small-molecule targeted therapies, immune checkpoint inhibitors, and tumor microenvironment modulation. Combination strategies incorporating next-generation cell therapies, oncolytic virus therapies, and therapeutic vaccines are also gaining prominence. Notably, clinical trials of personalized mRNA cancer vaccines have been promising, while antibody-drug conjugate and radionuclide-conjugated therapies present additional opportunities for enhancing AM prognosis. This article summarizes the cellular immune characteristics, mutation profiles, and tumor microenvironment of AM, as well as the current therapeutic strategies and advancements in the hope of expanding clinical benefits for AM patients.

Peptide nanomedicines have been rapidly developed in the field of tumor treatment and the discovery of new drugs due to their strong stability, precise targeting and high bioavailability. The administration methods of peptide nanomedicines include intravenous injection, oral administration, respiratory administration and transdermal administration, among which intravenous administration is the most commonly used. Once inside the body, peptide nanomedicines can be transported everywhere via the blood circulation, accumulating in large quantities at the target site. Effective drug tracking analysis methods can help to obtain accurate pharmacokinetic data, while a comprehensive understanding of the in vivo distribution and pharmacokinetic characteristics of peptide nanomedicines can contribute to the development of new drugs and promote the progress of cancer therapy. This paper reviews the advantages and disadvantages of analytical methods for peptide nanomedicines and the research progress in their metabolism and transport in order to provide references for subsequent research on peptide nanomedicines.

OBJECTIVE To explore the ameliorative effect of ursolic acid on carbon tetrachlorideinduced acute liver injury in mice, and the feasibility of multispectral optoacoustic tomography (MSOT) for characteristic structural and functional imaging of liver tissues. METHODS Kunming mice were randomly divided into the normal control group, model group, model+ursolic acid 30, 60 mg·kg^-1 groups and model+bifendate 5.625 mg·kg^-1 group, with 14 mice in each. Each group was given the corresponding drug once daily for 7 days. An acute liver injury model was established in mice by intraperitoneal injection of 0.2% carbon tetrachloride in olive oil solution after the last administration. Blood was collected, liver tissues were taken 24 h after modeling, and the liver index was calculated, 8 mice from each group and the levels of serum glutamic pyruciv transaminase (GPT) and glutamic oxaloacetic transaminase (GOT), as well as superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in liver tissues were measured. The enzyme-linked immunosorbent assay (ELISA) method was used to detect the level of α - glutathione S-transferase (α -GST) in serum. The histopathological changes of the liver were observed under a light microscope. The remaining 6 mice in each group underwent MSOT technique was used for characteristic structural and functional imaging of liver tissue. Levels of oxygenated hemoglobin (HbO₂) and deoxygenated hemoglobin (Hb) were analyzed, oxygen saturation was calculated, and the extent of liver injury was assessed by examining the intrahepatic distribution of indocyanine green (ICG). RESULTS Compared with the normal group, the levels of GPT, GOT and α-GST in serum, content of MDA in liver tissues and the liver index in the model control group were significantly increased while the activity of SOD in liver tissues were significantly decreased. Compared with the model group, ursolic acid in each dose group significantly reduced the liver index of mice, lowered the serum levels of GPT and GOT as well as the level of α- GST, decreased the content of MDA in liver tissues, and elevated the activity of SOD in liver - injured mice. Hematoxylin-eosin staining showed that significant steatosis and hepatocyte necrosis and inflammatory cell infiltration in hepatocytes of mice in the model group. Ursolic acid significantly attenuated the degree of hepatocellular lesions and markedly reduced steatosis in mice. MSOT imaging showed that the HbO2 level and oxygen saturation were significantly lower while the Hb level was remarkably higher in the liver of mice in the model group. In each administration group, the level of HbO₂ significantly increased, the level of Hb was significantly decreased, oxygen saturation was significantly increased in the liver of model mice and the accumulation of ICG dye probe was atten⁃ uated in the body after hepatocyte injury. CONCLUSION Ursolic acid can elevate the hepatic oxygen saturation, improve the metabolism of ICG, reduce the degree of hepatic necrosis in mice, and help protect against carbon tetrachloride-induced hepatic injury in mice. The mechanism is probably related to the inhibition of oxidative stress.

OBJECTIVE  To investigate the effect of N-methyl-D-aspartic acid (NMDA) receptor in the secondary visual cortex (V2) on methamphetamine-associated contextual learning and memory. METHODS  With male C57BL/6J mice as subjects and using the mouse conditioned place preference (CPP) experiment, the scores of CPP were observed after microinjection of NMDA receptor selective antagonist D-AP5 (0.5 μg per side) into the bilateral V2 during the formation phase, single microinjection of D-AP5 (0.5 μg per side) into the bilateral V2 prior to the expression test, and methamphetamine (0.5 mg∙kg^-1, ip)-induced reactivation test and methamphetamine-associated contextual-induced reactivation test, to evaluate the effect of NMDA receptors on the formation, expression and reinstatement of methamphetamine-induced CPP. RESULTS  Compared with the control group, microinjection of D-AP5 (0.5 μg per side) into the bilateral V2 during the formation phase did not have significant inhibitory effect on CPP scores, nor did single microinjection of D-AP5 (0.5 μg per side) into the bilateral V2 prior to the expression test, single microinjection of D-AP5 (0.5 μg per side) into the bilateral V2 prior to methamphetamine (0.5 mg∙kg-1, ip)-induced reactivation test or single microinjection of D-AP5 (0.5 μg per side) into the bilateral V2 prior to methamphetamine-associated contextual-induced reactivation test. CONCLUSION  The NMDA receptor in the V2 is not involved in the formation, expression and reactivation of methamphetamine-associated contextual learning and memory.

OBJECTIVE  To investigate the effects of ginsenosides as P-glycoprotein (P-gp) substrates in combination with paclitaxel on the proliferation and migration of colon cancer Caco-2 cells.  METHODS  Bio-layer interferometry (BLI) technology was used to detect the constants of ginsenosides and P-gp. Network molecular docking was adopted to predict the binding affinity energy of ginsenosides and P-gp. Caco-2 cells were divided into paclitaxel 0, 6.25, 12.5, 25, 50, 100 and 200 mg·L^-1 groups, ginsenoside Rg3 0, 6.25, 12.5, 25, 50, 100 and 200 mg·L^-1 groups, and paclitaxel 5 mg·L^-1+ginsenoside Rg3 0, 25, 50, 100 and 200 mg·L^-1 groups. After 48 h of incubation, the growth inhibition rate of Caco-2 cells was detected by MTT assay, and the interaction between the two drugs was quantitatively evaluated using the "one-belt, one-line" modle. Caco-2 cells were divided into the cell control group, paclitaxel 5 mg·L^-1 group, ginsenoside Rg3 50 and 100 mg·L^-1 groups, and paclitaxel 5 mg·L^-1+ginsenoside Rg3 50 and 100 mg·L^-1 groups. After 24 h of incubation, the proliferation and migration ability of the cells were detected by colony assay and Transwell migration assay. Caco-2 cells were then divided into the cell control group, quinidine 12.5 mg·L^-1 group, and ginsenoside Rg3 6.25 and 12.5 mg·L^-1 groups. After  4 h of incubation, the expression levels of P-gp and total protein were detected by ELISA. RESULTS  The affinity constants of ginsenoside Rb1, Rg3, Rg5 with P-gp were all less than 10^-3 mol·L^-1, while that of ginsenoside CK with P-gp was 10^-2 mol·L^-1. There was no typical binding dissociation curve between ginsenoside Re and P-gp. The absolute binding affinities of ginsenosides Rg3 and Rg5 to P-gp were determined to be 8.5 kcal·mol^-1 and 7.6 kcal·mol^-1, respectively. Ginsenosides mixed with PTX 5 mg·L^-1 inhibited the growth of colon cancer cells through synergy and addition, and the dose range of the synergistic effect was [0+5, 43.15+5] mg·L^-1; [164.51+5, 200+5] mg·L^-1, the additive effect dose ranged from [43.15+5, 164.51+5] mg·L^-1. The combination of the two drugs could significantly reduce the proliferation and migration ability of Caco-2 cells (P<0.01). The ELISA results showed a decrease in total protein and P-gp content in both the ginsenoside and quinidine groups (P<0.05). CONCLUSION  Ginsenoside bind to and inhibit the activity of P-gp, synergizing with paclitaxel to reduce the proliferative and migratory abilities of Caco-2 cells. The combination of ginsenosides and paclitaxel enhances the sensitivity of Caco-2 cells to paclitaxel induced inhibition. The combined use of these two substances is expected to achieve better anticancer effects compared to paclitaxel alone.

OBJECTIVE  To investigate the protective effect of interleukin-35 (IL-35) mRNA-lipid nanoparticles (LNP) against lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. METHODS  Fifity-six mice were randomly divided into 7 groups with 8 mice in each, including the normal control group, IL-35 mRNA-LNP (250 μg·kg^-1) group, LPS group, LPS+IL-35 mRNA-LNP (50,125 and 250 μg·kg^-1) group and LPS+Dexamethasone (DXM) group. Except for the normal control group and IL-35 mRNA-LNP (250 μg·kg^-1) group and ALI model was established by tracheal infusion of LPS in each of the other groups. IL-35 mRNA-LNP (250 μg·kg^-1) group and LPS+IL-35 mRNA-LNP (50,125 and 250 μg·kg^-1) group were injected with a corresponding dose of LNP encapsulated mRNA complex via the tail vein while the LPS+DXM group was injected with DXM via the tail vein. Lung coefficient and the wet to dry weight ratio (W/D) of lung tissue were recorded. The mRNA levels of inflammatory cytokines tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6  ) and IL-1β of lung homogenates were detected by real-time fluorescence quantitative PCR (RT-qPCR). LDH activity of lung homogenates and the protein levels of IL-35, TNF-α, IL-6 and IL-1β in lung homogenate were detected by corresponding kits. Hematoxylin-eosin (HE) staining was used to observe and analyze the pathological injury to lung tissue. The expression of Lymphocyte antigen 6G (Ly6G) was detected by Immunofluorescence to reflect the infiltration of neutrophils. RESULTS  Compared with the normal control group, LPS group and LPS+DXM group, IL-35 protein expression levels in lung homogenates of the other groups were more significant (P<0.01). Compared with the normal control group, lung coefficient, W/D ratio of lung tissue, LDH activity, mRNA levels and the protein levels of TNF-α, IL-6 and IL-1β in lung homogenates were significantly increased in the LPS group (P<0.01), accompanied by alveolar hemorrhage, alveolar wall thickening and neutrophils infiltration. After IL-35 mRNA-LNP administration, lung coefficient, W/D ratio of lung tissue, LDH activity, mRNA levels and the protein levels of TNF-α, IL-6 and IL-1β in lung homogenates were significantly decreased (P<0.01), and alveolar hemorrhage, alveolar wall thickening and neutrophil infiltration were obviously improved. CONCLUSION  IL-35 mRNA-LNP can express IL-35 protein in lung tissue of mice, and effectively improve LPS-induced ALI in mice by inhibiting the expression of proinflammatory factors.

OBJECTIVE  To investigate the roles and mechanisms of γ-bungarotoxin (γ-BGT) in inducing respiratory distress in mice. METHODS  Six male Kunming mice were selected and anesthetized before tracheal intubation and respiratory recording. After stabilizing respiration, the mice were intraperitoneally injected with γ-BGT at a dose of 6 mg·kg^-1. Once a decrease in respiratory frequency was observed, the mice were intravenously injected with nikethamide at a dose of 12.5 mg·kg^-1. Respiratory frequency was monitored using the BL420 signal acquisition and processing system. Male Kunming mice were randomly divided into the normal control group (saline, ip), γ-BGT group (6 mg·kg^-1, ip), and γ-BGT+nikethamide group (γ-BGT 6 mg·kg^-1, ip, followed by nikethamide 12.5 mg·kg^-1, ip, when shallow breathing and enhanced abdominal respiration were observed). The levels of Glu and GABA in the medulla oblongata were measured using ELISA. The protein expression levels of GAD65 and GAD67 in the medulla oblongata were determined by Western blotting. Primary mouse medullary neurons were cultured in vitro and divided into the following groups: cell control group, γ-BGT group, carbachol group, gallamine group, γ-BGT+H-89 group, and γ-BGT+Y-27632 group. The γ-BGT group, carbachol group, and gallamine group were incubated with γ-BGT (40 mg·L^-1), carbachol (100 mmol·L^-1), and gallamine (100 mmol·L^-1), respectively, for 4 h. The γ-BGT+H-89 and γ-BGT+Y-27632 groups were pretreated with γ-BGT (40 mg·L^-1) for 4 h, followed by incubation with the protein kinase A (PKA) inhibitor H-89 (50 mmol·L^-1) and the Ca²⁺ channel inhibitor Y-27632 (50 mmol·L^-1) for another 2 h, respectively. ELISA was used to measure the levels of Glu, GABA, cAMP, and calpain in the primary mouse medullary neurons. Western blotting was employed to assess the protein expression levels of GAD65 and GAD67, and PKA phosphorylation levels. Fluo-4 fluorescent probe was used to detect the intracellular Ca²⁺ level. RESULTS  The respiratory rate of mice significantly decreased after iv administration of γ-BGT (γ-BGT group) (P<0.05). After treatment with nikethamide (nikethamide group), the respiratory rate significantly recovered (P<0.05). Compared with the normal control group, the γ-BGT group exhibited a significant decrease in Glu content (P<0.05), a significant increase in GABA content (P<0.05), and a significant decrease in the Glu/GABA ratio. Additionally, the protein expression levels of GAD65 and GAD67 were significantly elevated (P<0.05). Compared with the γ-BGT group, the γ-BGT+nikethamide group showed a significant increase in Glu content (P<0.05), a significant decrease in GABA content (P<0.05), a significant increase in the Glu/GABA ratio, and a significant reduction in GAD65 and GAD67 protein expression levels (P<0.05). Compared to the cell control group, the γ-BGT group demonstrated a significant decrease in Glu content (P<0.05), a significant increase in GABA content (P<0.05), and a significant reduction in the Glu/GABA ratio. Furthermore, the protein expression levels of GAD65 and GAD67 were significantly elevated (P<0.05). Additionally, cAMP content, PKA phosphorylation levels, Ca²⁺ levels, and calpain activity were all significantly increased (all P<0.05). Glu, GABA, Glu/GABA ratio, and GAD expression levels in the γ-BGT group changed in the same way as in the gallamine group; In the γ-BGT+Y-27632 group, calpain activity and expression levels of GAD65 and GAD67 were all significantly decreased (all P<0.05). In the γ-BGT+H-89 group, Ca²⁺ levels and calpain activity were significantly reduced (all P<0.05). CONCLUSION  γ-BGT-induced poisoning can lead to respiratory distress in mice, possibly through the antagonism of M2 muscarinic acetylcholine receptors in medullary neurons, activation of the cAMP/PKA signaling pathway, elevation of intracellular Ca²⁺ levels, and increased expression and activity of GAD, resulting in an imbalance of Glu and GABA in the medulla.

OBJECTIVE  To investigate the effect and underlying mechanism of hydroxytyrosol (HT) on mouse chondrocyte injury induced by oxidative stress. METHODS  Mouse chondrocytes were incubated with varying concentrations of HT 0-400 μmol·L^-1 for 24 h, and the viability of the mouse chondrocytes was assessed using CCK-8 kit. An oxidative stress model of chondrocytes was established by the addition of H₂O₂ 200 μmol·L^-1. The experimental groups included the cell control group, H₂O₂ group, and H₂O₂+HT 10, 50 and 250 μmol·L^-1 groups. After 24 h, the mRNA expression levels of interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), prostaglandin E₂ (PGE₂), inducible nitric oxide synthase (iNOS), matrix metalloproteinase-3 (MMP-3 ), MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4 ), ADAMTS-5, SRY-box transcription factor-9 (SOX-9 ) and aggrecan (ACAN ) in mouse chondrocytes were detected by real-time quantitative PCR, the intracellular reactive oxygen species (ROS) level in chondrocytes was measured with 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA) staining, while the mitochondrial membrane potential was evaluated using JC-1 staining. After 48 h, the protein expression levels of iNOS, COX-2, MMP-13, and typeⅡcollagen (Col-2) in mouse chondrocytes were detected using Western blotting. RESULTS  HT at concentrations≤350 μmol·L^-1 had no significant effect on the survival of mouse chondrocytes. Compared with the cell control group, after 24 h, the mRNA expression levels of IL-6, COX-2, PGE2, iNOS, MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5 in the chondrocytes of mice in the H₂O₂ group were increased, while the mRNA expression levels of SOX-9 and ACAN were decreased. Additionally, there was an elevation in the ROS level and a significant loss of mitochondrial membrane potential in the chondrocytes of mice. Compared with the H₂O₂ group, after treatment with HT 10, 50 and 250 μmol·L^-1, there were significant decreases in mRNA expression levels of IL-6, COX-2, PGE2, iNOS, MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5, the mRNA expressions of SOX-9 and ACAN were increased, the ROS level was lowered. After treatment with HT 50 and 250 μmol·L^-1, the loss of mitochondrial membrane potential was ameliorated. Compared to the cell control group, the protein expressions of iNOS, COX-2 and MMP-13 were upregulated in the H₂O₂ group, while the protein expression of Col-2 was downregulated after 48 h. Compared to the H₂O₂ group, treatment with HT at concentrations of 10, 50 and 250 μmol·L^-1 resulted in decreased protein expressions of iNOS, COX-2 and MMP-13 in mouse chondrocytes, but the protein expression of Col-2 increased following treatment with HT 50 and 250 μmol·L^-1. CONCLUSION  HT can ameliorate H₂O₂-induced chondrocyte injury by reducing intracellular ROS levels and alleviating the loss of mitochondrial membrane potential, suppressing the release of inflammatory cytokines, inhibiting catabolic processes, and promoting anabolic activities.

As the longest and most widely distributed pair of nerves in the brain, the vagus nerve is involved in the regulation of many systems and organs. Recent studies have found that the vagus nerve may be involved in the occurrence of a variety of neuropsychiatric diseases by regulating the release of neurotransmitters (such as norepinephrine, 5-hydroxytryptamine, gamma-aminobutyric acid and acetylcholine) and regulating the immune system and gut-brain axis. This article focuses on the regulatory mechanisms of the vagus nerve on neurotransmitters, immune system function, and the gut-brain axis, as well the therapeutic advances in vagus nerve stimulation for neurological and psychiatric diseases such as epilepsy, depression and anxiety disorders. 

OBJECTIVE  To investigate the effect of dopamine receptor 3 (D₃R) on fear memory induced by intense electric shocks and the possible neurobiological mechanism. METHODS  ① To prevent pain threshold differences from influencing the effect of intense electric shocks, wild-type (WT) and D₃R knockout mice (D₃R^-/-) were used in the Hargreaves test to evaluate their basal pain threshold, with the paw withdrawal latency (PWL) as the observation index. ② WT and D₃R^-/- mice were divided into control groups and model groups, respectively. On the training day (the first day, D 1), the model groups received inescapable electric shocks (1.5 mA, 10 s, 10 s interval, 15 cycle) while the control groups did not. Contextual fear tests were conducted on D2, D7, D10, D14, and D16 after training, with the percentage of freezing time (FT) as the observation index to evaluate fear memory acquisition induced by contextual cues. On D17, after the model groups showed no more fear responses to contextual cues, they were re-stimulated with low-intensity current (0.5 mA, 10 s, 10 s interval, 15 cycle) to evoke fear memory. The two control groups did not receive any shocks. Contextual fear tests were conducted on day 18, and the FT% of each group was observed to evaluate fear memory retrieval induced by contextual cues. ③ Another cohort of WT and D₃R^-/- mice was used to further investigate the underlying neural mechanism, with the same grouping and treatment as in ②. Real-time dynamic changes in calcium signals of dopamine (DA) neurons in the ventral tegmental area (VTA) of WT and D₃R^-/- mice were detected using fiber photometry during electric shocks. The fluorescence area under the curve (AUC) was used as the indicator to quantify the excitability of DA neurons. RESULTS  ① In the Hargreaves test, there was no significant difference in PWL between D₃R^-/- mice and WT mice, indicating the two genotype mice had no significant differences in the basal pain threshold. ② Compared with the WT control group, the percentage of FT of the WT model group significantly increased on D2, D7, D10, and D14 (P<0.05). Compared with the D₃R^-/- control group, the percentage of FT of the D₃R^-/- model group significantly increased only on D2 and D7 (P<0.01). Meanwhile, the percentage of the FT of D₃R^-/- model group was significantly lower than in the WT control group on D2, D7, D10, and D14 (P<0.05, P<0.01). During memory recall (D18), the percentage of FT of the WT model and D₃R^-/- model groups significantly increased compared to their respective control groups (P<0.01, P<0.05), while the percentage of FT of D₃R^-/- model mice was significantly lower than that of WT model mice (P<0.01). ③ In the fiber photometry test, during the shock period, the calcium signals of DA neurons in the VTA of WT model and D₃R^-/-model mice rapidly increased within the first 2 s, and then gradually decreased between 2 to 10 s. The AUC within the 2 to 10 s interval was significantly lower in D₃R^-/- model mice compared to WT model mice (P<0.05), indicating that the excitability of DA neurons in the VTA of D₃R^-/-model mice was significantly lower than that of WT-model mice. CONCLUSION  D3R knockout inhibits the acquisition and recall of long-term fear memory in mice, and its neurobiological mechanism may be related to the decreased excitability of DA neurons during electric shock.

OBJECTIVE  To study the way in which hypidone hydrochloride (code: YL-0919) improves motor function after ischemic stroke (IS) and explore the related mechanism. METHODS  Adult male SD rats were used to establish a middle cerebral artery occlusion (MCAO) model that simulated acute IS. All animals were randomly divided into four groups: sham group, MCAO group, MCAO+YL-0919 group, and MCAO+YL-0919+erastin (Era, ferroptosis inducer) group. The drug administration groups received the first ip injection 6 h after operation, followed by continuous ip injection once per day. After 7-10 d of drug administration, the effect of YL-0919 on motor function after IS were evaluated via neurological function test, adhesive-removal test, rotarod test, balance beam test and open field test. After 7 d of drug administration, TTC staining was used to detect the cerebral infarction area while the colorimetry method was used to measure the contents of glutathione (GSH), malondialdehyde (MDA), and ferrous ions (Fe²⁺) in the penumbra of the cerebral cortex. Western blotting was used to detect the expression levels of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (xCT), acyl-CoA synthetase long-chain family member 4 (ACSL4), and transferrin receptor 1 (TFR1) in the cortical penumbra. RESULTS  Compared with the sham group, the MCAO group showed higher neurological function scores (P<0.01), with notably prolonged time for tape removal and first contact with the right forepaw (P<0.01), spent significantly more time crossing the balance beam (P<0.01) but endured a notably shorter duration on the rotarod (P<0.01), reduced the movement distance in the open field (P<0.01), had a remarkably increased infarct area (P<0.01) but significantly level of GSH in the cortical penumbra region decreased (P<0.01), while MDA and Fe²⁺ levels were markedly increased (P<0.01). Protein expression levels of GPX4 and xCT were reduced (P<0.05), while those of ACSL4 and TFR1 were elevated (P<0.05). Compared with the MCAO group, these changes were significantly reversed after YL-0919 administration. However, when Era and YL-0919 were administered simultaneously, the reversal effect of YL-0919 was significantly weakened. CONCLUSION  YL-0919 can improve motor function impairment and reduce cerebral infarction areas in rats after IS, and the mechanism may be related to the inhibition of ferroptosis.

OBJECTIVE  To establish a population pharmacokinetic (PopPK) model to predict the PK characteristics of GLS-010 in humans. METHODS  Fifty-eight cynomolgus monkeys were used, 18 of which were randomly divided into three groups and received a single intravenous infusion of GLS-010 at doses of 2, 6, and 18 mg·kg^-1, respectively. The rest were randomly assigned to four groups and received multiple intravenous infusions of GLS-010 at doses of 0, 5, 25, and 100 mg·kg^-1, respectively, once a week (quaque week, qw) for five consecutive weeks. Blood samples were collected before and after administration. The concentrations of GLS-010 in the monkey serum were measured using a validated enzyme-linked immunosorbent assay, while those of anti-drug antibodies (ADA) in the cynomolgus monkey serum were determined by ultra-sensitive electrochemiluminescence immunoassay. The PK data on GLS-010 in cynomolgus monkeys was obtained, and the drug-time curves were plotted. A PopPK model was constructed using non-compartmental analysis and evaluated by goodness-of-fit plots and visual predictive checks. The constructed PopPK model was used to predict the PK characteristics in humans, which were finally compared with actual PhaseⅠclinical study results for validation. RESULTS  The predictive results of the PopPK model were highly consistent with the actual PhaseⅠclinical study results. The model was able to predict the human PK characteristics under various dosing regimens, including 1 mg·kg^-1 quaque 2 weeks (q2w), 4 mg·kg^-1 (q2w), 240 mg (q2w), 240 mg (q3w), and 10 mg·kg^-1 (q2w). The predicted maximum plasma concentrations (C_max) were 24.8, 99.1, 85.0, 85.0, and 247.8 mg·L^-1, respectively, and the AUC_0-336 h was 4 902.0, 20 060.0, 17 147.7, 22 145.7 (AUC_0-504 h), and 50 817.6 mg·h·L^-1, respectively. The safety risks for the corresponding dosing regimens were 47.3, 11.6, 13.5, 10.5, and 4.6, respectively. The predicted receptor occupancy at steady state (RO_ss) at C_max, average plasma concentration (C_avg), and minimum plasma concentration (C_min) were 38.8%, 72.7%, 69.4%, 64.1% and 87.2%, 29.1%, 63.8%, 60.0%, 49.8% and 82.1%, 21.9%, 55.5%, 51.3%, 36.3% and 76.7%, respectively. CONCLUSION  The PopPK model can effectively predict the human PK characteristics under different dosing regimens with high consistency with actual PhaseⅠclinical study results, which can serve as an important reference for selection of safe and effective doses for first-in-human research.

Mesenchymal stem cells (MSCs) have become a key focus in the development of cell-based therapeutic products due to their wide availability, lack of ethical constraints, and potential for immune privileges. However, the transition from basic MSC transplantation techniques to fully developed therapeutic drugs presents numerous challenges given the current regulatory framework in China and from the perspective of drug development and review. This article summarizes the problems faced in this transition, particularly the challenges posed by theheterogeneity of MSC sources, the complexity of unique manufacturing processes, and the complexities of quality characterization. The article also offers suggestions and countermeasures in the hopes of advancing the research, development, and registration of MSC-based therapeutic products.

OBJECTIVE  To explore the possible molecular mechanism through which amygdala cell adhesion molecule-1 (CADM1) is involved in acute sleep deprivation-induced anxiety. METHODS  Sixteen 8-week-old C57BL/6J mice were randomly divided into the control group and para-chlorophenylalanine (PCPA)-induced acute sleep deprivation experimental group. The PCPA mice were intraperitoneally injected with PCPA suspension (at a dose of 300 mg∙kg^-1) between 8∶00 and 9∶00 am for 2 consecutive days while the control mice were injected with the same dose of normal saline. The sleep latency and sleep duration of mice were detected via the righting reflex test. Anxiety-like behaviors were detected by the open field test and elevated plus maze test. The expression level of CADM1 in the mouse amygdala was detected by Western Blot and Immunofluorescence staining. The GeneNetwork database was used to analyze the association between Cadm1 genes and other genes in the mouse amygdala. The key candidate regulatory genes were screened, and the Cadm1-anxiety behavior phenotype network was constructed. The mRNA expression levels of the key candidate regulatory genes were analyzed via qPCR analysis. RESULTS  Compared with the control mice, the sleep latency of PCPA mice was significantly prolonged (P<0.01) while the sleep duration was significantly shortened (P<0.01). The activity time and distance of PCPA mice in the open field center were significantly shorter than those of the control group (P<0.05). The elevated plus maze experiment showed that the percentage of the number of times PCPA mice entered the open arm and the percentage of residence time in the open arm were significantly lower than those of the control group (P<0.05). Western Blot and immunofluorescence staining showed that the expression of CADM1 protein in the amygdala from PCPA mice was down-regulated compared with the control mice (P<0.05). Based on gene-behavioral association network analysis, Cadm1 was significantly associated with 25 anxiety-like behavior-related genes. The enrichment analysis of Cadm1 co-expression genes showed that Cadm1 was associated with γ-aminobutyric acid GABAergic synaptic pathway (P=4.31e-09), and that the key genes were huntingtin associated protein 1 (Hap1) (r=0.705, P=1.09e-08)、inositol 1,4,5-triphosphate receptor type 1 (Itpr1) (r=-0.751, P =3.34e-10)、gamma-aminobutyric acid type A receptor subunit delta (Gabrd) (r=-0.836, P =3.93e-14)、γ-aminobutyric acid A receptor β1 subunit gene (Gabrb1) (r=0.732, P =1.50e-09) and adrenoceptor alpha 2A (Adra2a) (r=0.759, P=1.73e-10). The results of qPCR analysis showed that the mRNA levels of Hap1 (P<0.05)、Gabrb1 and Adra2a were significantly up-regulated (P<0.01) while those of Itpr1 and Gabrd were significantly down-regulated (P<0.01). CONCLUSION  Acute sleep deprivation leads to down-regulation of Cadm1 expression in the amygdala, and induces anxiety-like behaviors by affecting the expression of GABAergic synaptic signaling pathways Hap1, Gabrb1, Adra2a, Itpr1 and Gabrd.

OBJECTIVE  To investigate the protective effect of farrerol against lipopolysaccharide (LPS) induced acute lung injury (ALI) in mice and the mechanisms. METHODS  ① ICR rats were randomly  divided into the normal control group, model group and model+farrerol group, with 6 rats in each. The normal control group and model group were given an equal amount of normal saline intragaically for 4 d. The model+farrerol group was given 20 and 40 mg·kg^-1 intragaically for 4 d, while the normal control group was given an equal amount of normal saline intragaically on the 5^th day. The model group and model +farrerol group were injected with 3 mg·kg^-1 LPS solution for 24 h to construct an animal model of ALI. ② Mouse alveolar macrophages (MH-S) were cultured and randomly divided into the cell control group, model group and model+farrerol group.  Cell control group (conventional culture for 24 h), model group (100 μg·L^-1 LPS combined with 40 μg·L^-1 IFN-γ co-incubation for 24 h), model+farrerol group (5, 10 and 20 μmol·L^-1 farrerol pretreatment of MH-S cells for 24 h, and then 100 μg·L^-1 LPS combined with 40 μg·L^-1 IFN-γ co-incubation for 24 h), an inflammatory cell model was established in vitro. HE was used to observe the pathological changes of the lungs. The wet-dry weight ratio (W/D) was used to assess pulmonary edema while Evans blue dye (EBD) was used to detect pulmonary vascular permeability. The activity of myeloperoxidase (MPO) was detected by colorimetry. The number of white blood cells in BALF was counted with a blood cell counting plate. Cell proliferation assay (CCK-8) was used to determine the cell viability. The levels of interleukin-1β (IL-1β), tumor necrosis factor α (TNF-α), IL-6 and IL-10 in lung tissue, bronchoalveolar lavage fluid (BALF) and cell supernatants were detected via enzyme-linked immunosorbent assay (ELISA). The protein levels of inducable nitric oxide synthase (iNOS), arginase 1 (ARG1), phosphorylated NF-κB p65, NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase1 (caspase1) and gasdermin family member (GSDMD-N terminal) in lung tissue and MH-S cells were detected with Western blot. RESULTS  ① Compared with the normal control group, the histopathological changes, Injury score, W/D ratio, pulmonary vascular permeability, MPO content, leukocyte count, pro-inflammatory factor IL-1β, TNF-α, IL-6 levels, iNOS, phosphorylated NF-κB p65, NLRP3, caspase1 and GSDMD-N-terminal protein expression in lung tissues were significantly increased in the model group (P<0.05, P<0.01) while the expression levels of anti-inflammatory factors IL-10 and ARG1 were decreased (P<0.05, P<0.01). Compared with the model group, the Injury score, W/D ratio, pulmonary vascular permeability, leukocyte number, MPO content, proinflammatory factor content and iNOS, phosphorylated NF-κB p65, NLRP3, caspase1 and GSDMD-N-terminal protein levels were significantly decreased in the model+farrerol group(P<0.05, P<0.01) while the levels of anti-inflammatory factor IL-10 and ARG1 protein were increased (P<0.05,P<0.01). ② The results of in vitro experiments showed that compared with the cell control group, the contents of IL-1β, TNF-α and IL-6 and the expression levels of iNOS, phosphorylated NF-κB p65, NLRP3, caspase1 and GSDMD-N-terminal protein were increased (P<0.05, P<0.01), and that the content of anti-inflammatory factor IL-10 and expression level of ARG1 protein were decreased (P<0.01). Compared with the model group, the content of proinflammatory factor and the expressions of iNOS, phosphorylated NF-κB p65, NLRP3, caspase1 and GSDMD-N protein in the model+farrerol group were significantly decreased (P<0.05, P<0.01) while the expression levels of IL-10 and ARG1 protein were increased (P<0.05, P<0.01). CONCLUSION  Farrerol can alleviate acute lung injury induced by LPS in mice, possibly by inhibiting the phosphorylation of NF-κB p65 and activation of NLRP3 inflammatome, alleviating pyroptosis of cells and regulating macrophage polarization.

OBJECTIVE  To investigate the role and mechanism of microRNA-125b (miR-125b) in downregulating ion channel-related protein expression in a cardiac hypertrophy model. METHODS  ① In vivo:Lentiviral vectors for miR-125b overexpression and knockdown were constructed, and male C57BL/6 mice were divided into the following groups: sham group (thoracotomy without virus injection), LV-miR-125b group (mmu-miR-125b mimic), LV-miR-125b-inhibitor group (mmu-miR-125b-inhibitor), and negative control group (LV-NC). The mice were raised under normal conditions for 4 weeks. The ultrastructural changes in myocardium tissue sections of LV-miR-125b mice were observed using transmission electron microscopy. The cardiac hypertrophy model in mice was established using thoracic aortic constriction (TAC). Echocardiography was performed to measure ejection fraction (EF) and fractional shortening (FS), and the ratio of heart weight to body weight (HW/BW), ratio of heart weight to tibia length (HW/TL), as well as the expression level of the myocardial hypertrophy marker β-myosin heavy chain (β-MHC) were calculated to evaluate the success of the TAC-induced hypertrophy model. Subsequently, C57BL/6 mice were divided into four groups: Sham group, TAC model group, LV-miR-125b-inhibitor+TAC group, and LV-NC+TAC group. Protein expression levels of cardiac sodium channel (Nav1.5) and calcium channel (Cav1.2) were detected using Western blotting. RT-qPCR was performed to assess the levels of miR-125b and mRNA expression of myocardial hypertrophy markers, including atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and β-MHC. ②  In vitro: Primary cultured neonatal Kunming mouse cardiomyocytes were divided into four groups: cell control group (no treatment), miR-125b overexpression group, miR-125b-inhibitor group, and negative control group (NC). RT-qPCR was used to detect the levels of miR-125b,ANP,BNP, and β-MHC. Western blotting and immunofluorescence were performed to assess the expression levels of Nav1.5 and Cav1.2 in the cardiomyocytes. Luciferase reporter gene assay was used to evaluate the direct effect of miR-125b on the target proteins Nav1.5 and Cav1.2. RESULTS  ① In vivo: Compared to the Sham group, the TAC model mice showed significantly increased the ratio of heart weight to body weight (HW/BW), the ratio of heart weight to tibia length (HW/TL), and expression levels of the myocardial hypertrophy marker 
β-MHC (P<0.05), indicating the successful establishment of the TAC model. Furthermore, miR-125b expression was significantly elevated in the TAC model group (P<0.01). In the LV-miR-125b group, compared to the LV-NC group, the expression levels of myocardial hypertrophy markers ANP, BNP, and β-MHC were significantly increased (P<0.01), while the ejection fraction (EF) and fractional shortening (FS) values of the mice were significantly reduced (P<0.01). Additionally, Additionally, myocardium ultrastructure of LV-miR-125b group was damaged. Compared to the LV-NC+TAC group, the LV-miR-125b-inhibitor+TAC group showed a significant increase in ejection fraction (EF) and fractional shortening (FS) values (P<0.05). Additionally, the levels of Nav1.5 and Cav1.2 in myocardium tissue were significantly elevated in the LV-miR-125b-inhibitor+TAC group compared to the LV-NC+TAC group (P<0.05). ② In vitro: Compared to the NC group, the miR-125b overexpression group showed a significant 
increase in miR-125b expression (P<0.01), as well as elevated levels of ANP, BNP, and β-MHC (P<0.01). However, miR-125b-inhibitor significantly reversed the increases in ANP, BNP, and β-MHC (P<0.01). Western blotting and immunofluorescence results showed that, compared to the NC group, the miR-125b mimic group exhibited significantly decreased levels of Nav1.5 and Cav1.2 (P<0.01), while miR-125b-inhibitor led to an increase in the levels of both Nav1.5 and Cav1.2. Luciferase assay results demonstrated that miR-125b directly binds to the ion channel proteins Nav1.5 and Cav1.2, encoded by the SCN5A and CACNA1C genes. CONCLUSION  miR-125b promotes the development of cardiac hypertrophy by inhibiting the voltage-gated ion channel proteins Nav1.5 and Cav1.2 Inhibition of miR-125b expression improves cardiac hypertrophy.

OBJECTIVE  To study the pharmacokinetics/pharmacodynamics (PK/PD) profiles of meropenem in the plasma, lung, muscle and skin tissues of rabbits, and to explore the effects of sepsis and continuous renal replacement therapy (CRRT) on the probability of target attainment (PTA) of meropenem. METHODS Twenty-four New Zealand rabbits were randomly divided into four equal groups: sepsis-CRRT group, sepsis group, normal-CRRT group and normal control group. Six hours after a rabbit model of sepsis was established via cecal ligation and puncture (CLP) surgery, an automatic infusion pump with meropenem [(1 000/60)×3.27=54.5 mg·kg^-1, converted from that of humans (1 g) to that of rabbits] was used to administer the drug to the animals for 0.5 h, while CRRT was started simultaneously with drug administration in the CRRT group. Microdialysis samples were collected from 4 target sites of each group once every 30 min for 480 min. The concentration of meropenem was determined using liquid chromatography-tandem mass spectrometry (LC-MS/MS) before a concentration-time curve was constructed. Pharmacokinetic parameters such as the peak concentration (C_max), time to reach peak concentration (T_max), area under the concentration-time curve (AUC), and elimination half-life (t_1/2) were calculated using a non-atrioventricular model. Monte Carlo simulation (MCS) was performed at various minimum inhibitory concentrations (MICs) using pharmacokinetic parameters of meropenem to assess the probability of target attainment (PTA). The predefined PK/PD target was %fT>4MIC>40%, where the percentage of time that the free drug concentration [f] exceeded 4 times that of the MIC during a dosing interval was above 40% and C_max /MIC exceeded 4. RESULTS  The results indicated no statistically significant differences between the PK parameters of skin tissue in the normal control group and sepsis group, whereas C_max of other tissues was significantly lower in the sepsis group than in the normal control group (P<0.01). AUCplasma and AUCskin were significantly increased (P<0.01), while AUClung and AUCmuscle were significantly decreased (P<0.01) when the sepsis-CRRT group was compared with the sepsis group. With a target value of %fT>4MIC>40%, the plasma and skin tissues of the sepsis-CRRT group, the muscle tissues of the normal-CRRT group and the lung, muscle and skin tissues of the normal control group achieved a PTA of over 90% (MIC=1 mg·L^-1). The skin tissues of the sepsis-CRRT group and the lung tissues of the normal control group achieved an efficacy target of more than 90% (MIC=2 mg·L^-1). However, our results did not indicate that either AUC or C_max was involved in the changes of PTA in the normal control group. For all tissues except skin tissues (MIC=4 mg·L^-1), PTA decreased with C_max in the sepsis group. The PTA of plasma and skin tissues increased with the AUC, while the PTA of lung (MIC=2, 4 mg·L^-1) and muscle tissues (MIC=1, 2, 4 mg·L^-1) decreased with the AUC in the sepsis-CRRT group. CONCLUSION  Sepsis has little effect on the distribution of meropenem in skin tissues. Sepsis may reduce the efficacy of meropenem in various tissues, and CRRT can improve the efficacy of plasma and skin tissues in sepsis. This method of collecting target samples with microdialysis probes is accurate and reliable, which can be used in therapeutic drug monitoring.

Zinc oxide nanoparticles (ZnO NPs) have been gradually introduced into people′s lives due to their excellent properties. In daily contact, ZnO NPs can enter the human body in a variety of ways and accumulate in tissues and organs. Inhalation is an important means by which ZnO NPs enter the human body and deposit in lungs. Studies have shown that exposure of lungs to ZnO NPs will cause a range of negative effects. The mechanism of pulmonary toxicity induced by ZnO NPs is related to Zn2+ release, oxidative stress, DNA damage, autophagy, inflammation and pulmonary surfactant 
dysfunction. This paper reviews the pulmonary toxicity and mechanisms of ZnO NPs, and summarizes the prevention and treatment strategies, which provides scientific basis for safer use of nanomaterials, and potential intervention strategies for the prevention and treatment of nanotoxicity.

Drug addiction is a major medical and social issue globally, but there is still a lack of ideal clinical treatments. Glucagon-like peptide-1 (GLP-1), a key gut-brain peptide, and its receptor agonists have been approved for the treatment of type 2 diabetes and obesity. In recent years, the role of GLP-1 receptor agonists in combating drug addiction has received increasing attention. Based on an overview of the GLP-1 system and its receptor agonist, this article reviews the role of GLP-1 receptor agonists in drug addiction at both the nonclinical and clinical levels by focusing on addictive psychoactive substances (including alcohol, tobacco, cocaine, opioids, and amphetamines). Future developments are also predicted to provide new clues to the mechanism of drug addiction and development of therapeutic drugs.

Ligand-gated ion channels transduce chemical signals into ionic currents while being bonded with specific ligands, thereby facilitating signal transduction and intercellular communication. These channels have become critical therapeutic targets for a variety of diseases. Ginsenosides, triterpenoid saponins primarily derived from herbs of the Panax genus, are known for their broad range of biological activities. Ginsenosides and their metabolites can regulate the functions of various ligand-gated ion channels, including nicotinic acetylcholine receptors, 5-hydroxytryptamine type-3 receptors, γ-aminobutyric acid type-A receptors, glycine receptors, ionotropic glutamate receptors, ATP-gated P2X channels, and transient receptor potential channels. They play important roles in anti-inflammation, analgesics, immune regulation, anti-epilepsy, cognitive enhancement and neuroprotection. Further investigation into the mechanisms by which ginsenosides regulate these channels will provide important evidence for their clinical applications and the development of new drugs.

OBJECTIVE  To investigate the effects of potassium channel Kv1.3 knockout (Kv1.3 KO) on neurological dysfunction and neuroinflammation in C57BL/6 mice following traumatic brain injury (TBI).  METHODS  C57BL/6 mice and homozygous Kv1.3 KO C57BL/6 mice were subjected to the classic controlled cortical impact model to establish a TBI model. The experimental groups included the sham surgery group, C57BL/6 TBI model group (TBI group), and a Kv1.3 KO C57BL/6 TBI model group (TBI+Kv1.3 KO group). At 1, 2, and 3 weeks post-modeling, real-time quantitative PNCR was used to measure the mRNA expression levels of Kv1.3, interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TF-α), and IL-10 in hippocampal tissues. At 1 and 3 weeks post-modeling, Western blotting was performed to detect Kv1.3 protein expressions in the hippocampus. At 3 weeks post-modeling, Western blotting was used to assess the protein levels of IL-1β, IL-6, TNF-α, and IL-10 in hippocampal tissues. Additionally, immunofluorescence was employed to quantify cells co-labeled with the microglial marker ionized calcium-binding adapter molecule 1 (IBA1) and Kv1.3, IL-1β, or TNF-α in the hippocampus. Patch-clamp recordings were conducted to measure Kv1.3 channel currents in primary microglia at 3 weeks post-modeling. Neurological function was evaluated at 1 and 3 weeks post-modeling using the neurological severity score (NSS), pole climbing, and rotarod tests. Cognitive function was assessed at 3 weeks post-modeling via open field, Morris water maze, and Y-maze tests. RESULTS  Compared with the sham group, the TBI group exhibited significantly elevated mRNA expression levels of Kv1.3 and IL-1β in the hippocampus at 1, 2 and 3 weeks post-modeling, while IL-6 and IL-10 mRNA levels showed no significant changes. Notably, TNF-α mRNA expressions demonstrated a significant increase only at 2 and 3 weeks post-modeling. At 1 and 3 weeks post-modeling, Kv1.3 protein expressions in the hippocampus were significantly higher in the TBI group. At 3 weeks post-modeling, hippocampal IL-1β and TNF-α protein levels were markedly increased in the TBI group, whereas IL-6 and IL-10 protein levels did not change significantly. Moreover, Kv1.3 current density in primary microglia was significantly enhanced in the TBI group at 3 weeks post-modeling. Immunofluorescence analysis revealed that the number of IBA1-positive microglia co-labeled with Kv1.3, IL-1β, or TNF-α in the hippocampus was significantly larger in the TBI group than in the sham group at 3 weeks post-modeling. Behaviorally, the TBI group exhibited significantly higher NSS scores, lower success rates in full turn attempts, and longer times taken to descend the pole at 1 and 3 weeks post-modeling compared with the sham group. At 3 weeks post-modeling, TBI mice also demonstrated reduced total movement distance in the open field, decreased time spent in the central zone, fewer platform crossings, less time in the target quadrant, and lower spontaneous alternation rates. In contrast, the TBI+Kv1.3 KO group showed significantly improved outcomes compared with the TBI group: lower NSS scores, higher success rates in full turns, and shorter time taken to descend the pole at 1 and 3 weeks post-modeling. At 3 weeks post-modeling, the TBI+Kv1.3 KO group displayed longer rotarod endurance, increased total movement distance in the open field, more time spent in the central zone, higher platform crossings, greater target quadrant exploration time, and improved spontaneous alternation rates. Furthermore, at 1 and 3 weeks post-modeling, the TBI+Kv1.3 KO group exhibited significantly reduced mRNA expression levels of the 
inflammatory cytokines IL-1β and TNF-α in the hippocampus compared with the TBI group. CONCLUSION  Potassium channel Kv1.3 knockout mitigates neurological dysfunction and neuroinflammation in C57BL/6 mice following TBI. 

OBJECTIVE  To investigate the effect and mechanism of soluble guanylate cyclase stimulator sGC003F on cardiac function in mice with chronic heart failure (CHF). METHODS  C57BL/6J male mice were randomly divided into the sham operation (sham) group, transverse aortic constriction induced CHF mouse model group, model+veliciguat (Ver, 3 mg·kg^-1) group (positive control) and model+sGC003F (3 and 10 mg·kg^-1) group. Four weeks after modeling, drugs were ig given, once a day, for 28 d. Echocardiography was used to measure the changes in cardiac function, and the myocardial hypertrophy related indexes were calculated. The levels of serum N-terminal pro-brain natriuretic peptide (NT-pro-BNP), N-terminal pro-atrial natriuretic peptide (NT-pro-ANP), soluble guanylate cyclase (sGC), cyclic guanosine monophosphate (cGMP) and inflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and IL-1β were detected by ELISA. The pathological changes of left heart tissue were observed with HE and Masson staining. Image was used to analyze the percentage of fibrosis in cardiac tissus stained with Masson. The activity of superoxide dismutase (SOD), content of malondialdehyde (MDA) in myocardial tissue, and level of nitric oxide (NO) in serum were detected by biochemical detection kits. The protein expression levels of p-mammalian target of rapamycin (p-mTOR), p-protein kinase B (p-Akt), TNF-α and IL-6 in cardiac tissue were detected by Western blotting. RESULTS  Compared with the sham group, the left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFs) in the model group decreased significantly (P<0.01), the cardiac structure changed significantly, the percentage of myocardial fibrosis increased significantly (P<0.05), so were serum NT-pro-BNP and NT-pro-ANP levels (P<0.01). Compared with the model group, the above indexes of the model +Ver group and the model+sGC003F 3 mg·kg-1 group were significantly improved (P<0.05, 
P<0.01). The sGC003F 10 mg·kg-1 group had a significant improvement in LVEF, LVFs, and NT-pro-BNP (P<0.01). Compared with the sham group, the serum levels of NO, sGC and cGMP in the model group decreased significantly (P<0.05, P<0.01). Compared with the model group, the serum levels of NO, sGC and cGMP were significantly increased in the model+sGC003F 3 mg·kg^-1 group (P<0.01), but only serum cGMP levels were significantly increased in model +Ver and model+sGC003F 10 mg·kg^-1 groups (P<0.01).Compared with the sham group, the serum levels of TNF-α, IL-1β and IL-6 in the model group were significantly increased (P<0.05, P<0.01). Compared with the model group, the serum levels of TNF-α, IL-1β and IL-6 were significantly decreased in the model+sGC003F 3 mg·kg^-1 group (P<0.05, P<0.01), and only the TNF-α level was significantly decreased in the model+sGC003F 10 mg·kg^-1 group (P<0.01). Compared with the sham group, the SOD activity of the model group was significantly decreased (P<0.01), but the MDA content significantly increased (P<0.01). Compared with the model group, SOD and MDA were significantly improved in the model+sGC003F 3 mg·kg^-1 group (P<0.05, P<0.01), but in the model+Ver group only the SOD activity significantly increased (P<0.05). Western blotting showed that the expressions of p-mTOR, p-Akt, TNF-α and IL-6 protein in myocardial tissue of the model group were significantly higher than in the sham group (P<0.05). Compared with the model group, the expressions of the above proteins in the model+sGC003F 3 mg·kg^-1 group were significantly decreased (P<0.05, P<0.01), so were the expressions of TNF-α protein in the model+sGC003F 10 mg·kg^-1 group and model+Ver group (P<0.01). CONCLUSION  sGC003F can improve cardiac function, and reduce myocardial fibrosis in  CHF model mice, which may be related to the inhibition of myocardial oxidative stress and inflammation, and the regulation of NO/sGC/cGMP and AKT/mTOR signaling pathways.

OBJECTIVE  To study the effects of SR9009 and LXH0225, nuclear receptor subfamily 1 group D member (REV-ERB) agonists,  on mood disorders and cognitive impairment in over-training mice. METHODS  Male C57BL/6J mice were randomly divided into the normal control, over-training model, model+fluoxetine (15 mg∙kg^-1), model+SR9009 (100 mg∙kg^-1) and model+LXH0225 (50 mg∙kg^-1) groups. Mice in the normal control group were ip given15% cremophor without extra stress while those in other groups were ip given 15% cremophor or different drugs respectively 30 min before daily forced swimming stress. When stressed, mice were forced to swim in 19-21 ℃ water for 20 min per day for 18 d. After that, locomotor activity was assessed. Rotarod test and weight-loaded swimming test were performed to measure physical strength, while open field test and stair-climbing test were performed to measure anxiety-like behavior. Tail suspension test and forced swimming test were used to measure depression-like behavior while novel object recognition test and Y maze test were conducted to measure recognition function. ELISA was used to measure serum corticosterone contents. RESULTS  Compared with the normal control group, the locomotor activity of mice in the model group was significantly increased (P<0.01). There were anxiety-like behaviors with a significant increase in the number of times of stair-climbing and rearing (P<0.01). Depression-like behaviors were observed with a significant increase immobile time in forced swimming test (P<0.01). Cognitive impairment was manifested as decreased accuracy of Y-maze spontaneous alternation response (P<0.01). The corticosterone content was significantly elevated (P<0.01) in forced swimming mice. Compared with the model group, the accuracy of Y-maze spontaneous alternations was higher (P<0.01) and the content of serum corticosterone was lower (P<0.01) in the model+fluoxetine group. The immobile time was shorter (P<0.01), the  spontaneous alternation response of Y maze was was less accurate (P<0.05) and serum corticosterone content was lower (P<0.01) in the model+SR9009 group than in the model group. The latency to fall off the rotarod was longer (P<0.05), the immobile time was shorter (P<0.01) and the content of serum corticosterone was lower (P<0.01) in the model+LXH0225 group compared with the model group. CONCLUSION  The REV-ERBs agonists SR9009 and LXH0225 may protect against forced swimming over-training induced mood disorders and cognitive impairment. 

OBJECTIVE  To investigate the regulatory effects of salvianolic acid C (SAC) on the level of cuproptosis and inflammatory injury in cardiomyocytes after myocardial infarction (MI). METHODS  ① C57BL/6 mice were divided into a sham group, an MI model group, and SAC (5, 10 and 20 mg·kg^-1) groups, with 10 mice in each group. Mice in the SAC groups were pretreated with oral gavage of SAC for 1 week, while those in the sham and model groups received an equal volume of saline. One week later, an MI model was established in the model and SAC groups by ligating the left anterior descending coronary artery, while the sham group underwent thoracotomy without ligation. MI size was assessed using triphenyltetrazolium chloride (TTC) staining. Cardiomyocyte apoptosis was evaluated by TUNEL staining. The ultrastructure of cardiomyocyte mitochondria was observed under a transmission electron microscope. ② Mouse cardiomyocytes HL-1 were divided into a control group, an oxygen-glucose deprivation (OGD) model group, OGD+SAC 1, 5 and 10 μmol·L^-1  groups, and a OGD+SAC (5 μmol·L^-1)+nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor ML385 (2 μmol·L^-1) group. Cells in the OGD+SAC groups were pretreated with SAC for 24 h while those in the OGD+SAC+ML385 group were pretreated with both SAC 5 μmol·L^-1 and ML385 2 μmol·L^-1 for 24 h. Except for the control group, an OGD model was established in HL-1 cells. ELISA was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in mouse serum and HL-1 cell culture supernatants. The Cu⁺ detection kit was used to measure Cu⁺ levels in myocardial tissue and HL-1 cells. Cell viability was assessed using the CCK-8 kit. Apoptosis rates of HL-1 cells were detected by flow cytometry. Reactive oxygen species (ROS) levels in HL-1 cells were measured using a ROS detection kit. Western blotting analysis was performed to detect the expression levels of Nrf2, heme oxygenase-1 (HO-1), and cuproptosis markers, ferredoxin 1 (FDX1) and solute carrier family 31 member 1 (SLC31A1) in myocardial tissue and HL-1 cells. RESLUTS  ① Compared with the sham group, the MI model group exhibited increased myocardial infarction size, elevated cardiomyocyte apoptosis rates, mitochondrial swelling, vacuolation, and cristae rupture in cardiomyocytes, increased serum levels of TNF-α, IL-6, and IL-1β, elevated Cu+ levels and expressions of FDX1 and SLC31A1 in myocardial tissue, and decreased expressions of Nrf2 and HO-1 (P<0.01). Compared with the model group, the SAC 5, 10 and 20 mg·kg^-1 groups showed 
reduced MI size, decreased cardiomyocyte apoptosis rates, alleviated mitochondrial swelling, vacuolation, and cristae rupture, lower serum levels of TNF-α, IL-6 and IL-1β, decreased Cu⁺ levels and expressions of FDX1 and SLC31A1 in myocardial tissue, and increased expressions of Nrf2 and HO-1 (P<0.05, P<0.01). ② Compared with the cell control group, the OGD model group demonstrated significantly decreased HL-1 cell viability, increased cell apoptosis rates, Cu⁺ and ROS levels, expressions of FDX1 and SLC31A1, elevated levels of TNF-α, IL-6 and IL-1β in cell culture supernatants, and 
decreased expressions of Nrf2 and HO-1 (P<0.01). Compared with the OGD model group, the SAC 1, 5 and 10 μmol·L^-1 groups showed increased HL-1 cell viability, decreased cell apoptosis rates, Cu⁺ and ROS levels, expressions of FDX1 and SLC31A1, reduced levels of TNF-α, IL-6 and IL-1β in cell culture supernatants, and increased expressions of Nrf2 and HO-1 (P<0.05, P<0.01). Compared with the SAC 5 μmol·L^-1 group, the SAC 5 μmol·L^-1+ML385 2 μmol·L^-1 group exhibited decreased cell viability, increased cell apoptosis rates, Cu⁺ and ROS levels, expressions of FDX1 and SLC31A1, elevated 
levels of TNF-α, IL-6, and IL-1β in cell culture supernatants, and decreased expressions of Nrf2 and HO-1 (P<0.01). CONSLUSION  SAC can activate the Nrf2/HO-1 signaling pathway, alleviate cuproptosis in cardiomyocytes after MI, and reduces inflammatory damage.