Weichang'an pill (WCAP) is a traditional Chinese medicine formulation commonly used in the treatment of functional digestive disorders. Despite its widespread clinical application and demonstrated efficacy, its in vivo chemical composition and pharmacokinetic characteristics remain insufficiently elucidated. This study firstly employed a UPLC-Q Exactive Focus-MS/MS system to analyze its prototype components in the plasma and tissues (including the stomach, small intestine, liver, and kidney) of SD rats. Following administration, 38 prototype components were detected in rat plasma and tissues, including 10 flavonoids, 6 phenylpropanoids, 6 2-(2-phenylethyl)chromones, 5 phthalides, and 4 terpenoids, along with other compounds. Among these, 23 prototype components were detected in plasma, while 38, 38, 34, and 32 prototype components were identified in the stomach, small intestine, liver, and kidney, respectively. Furthermore, a quantitative analytical method using UPLC-QqQ-MS/MS was developed for 15 major components in WCAP. Results indicated that WCAP contained relatively high levels of naringin, honokiol, and magnolol, at 4. 85 mg/g, 5. 11 mg/g, and 6. 44 mg/g respectively. Therefore, the pharmacokinetics of 8 WCAP constituents (agarotetrol, senkyunolide A, butylphthalide, dehydrocostus lactone, naringin, rhein, honokiol, and magnolol) were evaluated in rats after oral administration. Pharmacokinetic results indicate that the T1/2 of all 8 components exceeded 5 h, while the MRT(0-∞) exceeded 10 h. With the exception of agarotetrol and dehydrocostus lactone, the Tmax of the remaining 6 components was less than 0. 25 h. In conclusion, this paper researched the chemical constituents in vivo and pharmacokinetics of WCAP, which provided the reference for further study.

Methamphetamine (METH) is highly addictive and can cause neurotoxicity when misused over a prolonged period. METH-induced neurotoxicity and addiction involve multi-level biological alterations, including nucleotide polymorphisms, aberrant epigenetic modifications, transcriptional imbalance, protein dysfunction, metabolic dysregulation, and microbial dysbiosis with changes in microbiota-derived metabolites. To systematically dissect these multifaceted changes, genomics has been deployed to identify key gene polymorphisms linked to METH addiction susceptibility and epigenetic changes such as DNA methylation in brain regions following METH exposure. Tran-scriptomics dynamically profiles differential gene expression in addiction-relevant brain areas, particularly in pathways governing synaptic plasticity and neuroinflammation. Proteomics pinpoints the dysregulation of functional proteins associated with synaptic plasticity in response to METH. Metabolomics quantifies neurotransmitter depletion and redox imbalance induced by the drug, while microbiomics reveals gut microbiota dysbiosis and subsequent neural damage via the gut-brain axis. The integrated application of these multi-omics technologies-spanning genetic variation, transcriptional regulation, protein function, metabolic dynamics, and host-microbe interactions-systematically illuminates the complex molecular events underlying METH neurotoxicity and addiction, thereby providing a robust theoretical framework and technical roadmap for identifying early warning biomarkers and devising multi-target combination strategies against METH neurotoxicity and relapse. This review summarizes the current state of single-omics and multi-omics applications in METH neurotoxicity and addiction research, aiming to provide a reference for elucidating pathogenic mechanisms, identifying biomarkers, and discovering potential therapeutic targets.

Methamphetamine (METH) abuse and HIV infection have emerged as two major public health concerns worldwide. Both METH and the HIV-1 Tat protein (Tat) can enter central nervous system through blood-brain barrier (BBB) and exert neurotoxic effects. However, the specific mechanism underlying their synergistic disruption of BBB remains unclear. Herein, co-culture models of endothelial cells and astrocytes and tree shrews were used to explore the role and mechanism of the Transient receptor potential M2 (TRPM2) in mediating the synergistic BBB damage induced by METH and Tat. In the tree shrew experiment, METH and/or Tat exhibited neurotoxicity, and the combination of them yielded more severe toxic effects characterized by tight junction protein loss, albumin leakage, astrocyte activation, neuroinflammation, and astrocytic TRPM2 activation. Similarly, in vitro experiments revealed that METH and Tat decreased tight junction proteins and transendothelial electrical resistance while increasing the flux of sodium fluorescein, and triggering astrocyte-mediated inflammation and astrocytic TRPM2 activation. However, pharmacological inhibition of TRPM2 mitigated the above-mentioned BBB damage and astrocyte-related inflammation both in vitro and in vivo. Furthermore, inhibition of TRPM2 mitigated METH + Tat-induced stereotypical behavior and anxiety-like behavior in tree shrews. Critically, astrocyte-specific TRPM2 knockdown in tree shrews effectively prevented METH and Tat-induced neuroinflammation and BBB damage, and ameliorated the associated behavioral impairments. This study revealed that METH and Tat could synergistically activate astrocytic TRPM2 channels to induce astrocyte-derived inflammatory response, and ultimately resulted in BBB injury. These findings highlight astrocyte-mediated inflammation as an important mechanism by which METH and Tat synergistically damage the BBB, and the TRPM2 channel represents a promising therapeutic target, providing novel insights into the mechanism of neurotoxicity caused by METH and HIV-1 Tat.

BackgroundWhile Western medicine is increasingly investigating sex- and gender-specific differences in health and disease, and despite TCM theory and practice placing particular emphasis on individual differences—including sex- and gender-related characteristics, research on sex- and gender- sensitive medicine in TCM remains underexplored.ObjectiveWe conducted a scoping review to map and synthesize the existing evidence on sex- and gender-specific differences in the efficacy and safety of TCM interventions and to analyze possible underlying mechanisms.MethodsThe scoping review followed the JBI guidance. Six Chinese and English databases (CNKI, WF, VIP, SinoMed, PubMed, and Web of Science) were searched from inception to July 2025 for preclinical and clinical studies. Key information was extracted from the included studies, and data were synthesized using descriptive statistics and systematic narrative synthesis. GraphPad Prism 10.0 was used to generate figures.ResultsDatabase searches identified 6,720 records; after removing 250 duplicates, 6,470 records were screened by title and abstract, and 2,417 proceeded to full-text eligibility assessment. Ultimately, 16 studies were included, comprising randomized controlled trials, animal experiments, non-randomized controlled trials, and single-arm trials. Conclusions drawn from the original studies indicated that changes in all relevant outcome measures were associated with sex. Sex-specific differences in TCM efficacy were observed across herbal, acupuncture, and combined interventions. Female animals demonstrated predominant benefits in bone metabolism, whereas female human participants showed greater benefits in pain perception, emotional regulation, neurological recovery, and pulmonary functional outcomes. Male animals exhibited more pronounced gut microbiota responses to metabolic interventions, while male human participants showed stronger effects in obesity-related outcomes and sensorimotor responses. These divergent patterns likely reflect sex-related differences in pharmacokinetics, gut-brain axis interactions, and neuroendocrine mechanisms. None of the included studies examined gender-related sociocultural factors.ConclusionThis review highlights a persistent theoretical and methodological gap in TCM research, which has yet to fully integrate sex- and gender-sensitive medicine principles. The limited high-quality clinical evidence underscores the need for rigorous sex-informed trials to evaluate efficacy across sexes. These findings support the integration of sex as a key determinant in TCM clinical decision-making, alongside syndrome differentiation, to advance precision medicine and improve personalized therapeutic outcomes.Systematic Review RegistrationDOI: 10.17605/OSF.IO/QABTY.

Methamphetamine (Meth) is known to cause cognitive impairment in drug abusers, but the underlying mechanisms remain unclear. Given that Meth is also an emerging environmental contaminant detected in surface waters, understanding its neurotoxic mechanisms has implications for health risk assessment. Melatonin, a natural neuroprotective agent, shows promise in alleviating cognitive deficits, but its mechanism of action in Meth exposure is unclear. We found that melatonin treatment alleviated Meth-induced cognitive deficits, microglial activation, and synaptic damage, while also mitigating colonic inflammation and altering gut microbiota composition. Antibiotics treatment and fecal microbiota transplantation experiments validated the indispensable role the gut microbiota in melatonin's mitigation of Meth-induced cognitive impairment. Mechanistically, melatonin significantly elevates levels of the tryptophan metabolite 3-hydroxyanthranilic acid (3-HAA) by reshaping the gut microbiota. Exogenous supplementation of this compound reproduces melatonin's protective effects. Combining proteomic and transcriptomic analyses, we found that 3-HAA downregulates NLRP3 expression in hippocampal microglia to suppress neuroinflammation and synaptic damage, thereby alleviating Meth-induced cognitive impairment. Conversely, overexpression of NLRP3 abolishes the protective effects of 3-HAA. Collectively, our findings uncover a gut‑brain axis mechanism by which melatonin counteracts Meth‑induced neurotoxicity, providing a potential therapeutic target and informing the health risk assessment of Meth as an environmental contaminant.

To investigate the therapeutic effect of Kushen Tongguan Pill (KSTG) on benign prostatic hyperplasia (BPH) induced by castration combined with testosterone propionate in rats, and to explore its underlying mechanism associated with the Toll-like receptor 4/nuclear factor κB (TLR4/NF-κB) signaling pathway. A rat BPH model was established by castration combined with subcutaneous injection of testosterone propionate. Rats were randomly divided into sham group, BPH group, and low-, medium-, and high-dose KSTG groups (2.52, 5.04, and 10.08 g/kg). After 4 weeks of intervention, prostate wet weight was weighed and prostate index was calculated； histopathological changes were observed by hematoxylin-eosin (HE) staining; collagen deposition and fibrosis were evaluated by Masson’s trichrome staining; the expression of α-smooth muscle actin (α-SMA) was detected by immunohistochemistry; serum testosterone (T), estradiol (E2), and dihydrotestosterone (DHT) levels were measured by enzyme-linked immunosorbent assay (ELISA); peripheral white blood cell and neutrophil counts were measured using an automated hematology analyzer; toluidine Blue staining was performed to observe mast cell infiltration in prostate tissue to assess local chronic inflammation. RT-qPCR was used to detect the mRNA expression of key pathway molecules including TLR4, NF-κB p65, IKKα, and IKKβ, as well as inflammatory markers TNF-α, IL-1β, MCP-1, and iNOS in prostate tissue. Western blotting was performed to analyze the protein expression of key pathway molecules including TLR4, MyD88, IκBα, p-IκBα, NF-κB p65, and p-NF-κB p65, as well as the inflammatory marker iNOS in prostate tissue. For in vitro experiments, KSTG-containing serum was used to treat LPS/IFN-γ-induced THP-1-derived macrophages. The mRNA expression of M1 polarization markers (CD86, TNF-α, IL-6) in macrophages was determined by RT-qPCR. Subsequently, the conditioned medium from each macrophage group was collected, and its effect on WPMY-1 cell proliferation was evaluated by CCK-8 assay. Compared with the BPH group, KSTG dose-dependently reduced prostate volume, wet weight, and prostate index (P < 0.05) and improved the pathological morphology of glandular hyperplasia and interstitial thickening. KSTG dose-dependently reduced serum E2 levels and the E2/T ratio, and significantly decreased the abnormally elevated serum DHT levels (P < 0.05). KSTG also reduced peripheral white blood cell and neutrophil counts, downregulated the mRNA expression of TNF-α and IL-1β, and decreased the protein expression of iNOS in prostate tissue (P < 0.05). Toluidine blue staining further revealed that KSTG treatment reduced mast cell infiltration in the prostate, corroborating the alleviation of local inflammation (P < 0.05). Western blotting showed that KSTG significantly inhibited the protein expression of TLR4 and MyD88 in prostate tissue and suppressed the phosphorylation of IκBα and NF-κB p65 (P < 0.05). In vitro, KSTG-containing serum inhibited the mRNA expression of M1 polarization markers CD86, TNF-α, and IL-6 in THP-1-derived macrophages (P< 0.05). Moreover, the conditioned medium of macrophages pretreated with KSTG significantly attenuated the pro-proliferative effect on WPMY-1 cell proliferation (P < 0.05). This study has demonstrates that the KSTG Pill can significantly regulate M1 macrophage polarization to alleviate the prostatic hyperplasia and related hormonal level in the BPH rats. Its therapeutic mechanism is closely related to suppression of aberrant activation of the TLR4/MyD88/NF-κB signaling pathway in the prostate tissue, thereby reducing the polarization of macrophages to the M1 phenotype and the release of downstream pro-inflammatory factors. These findings provides experimental support for its clinical application, and and valuable insights for the development of novel BPH treatment strategies targeting the immune-inflammatory microenvironment.

Mitochondria are metabolic hubs that house their own genomes (mitochondrial DNA [mtDNA]), which encode components of the oxidative phosphorylation (OXPHOS) machinery. The mitochondrial central dogma not only governs compartmentalised metabolism but also intensively intertwines with multiple biological processes, and its dysregulation is a hallmark of cancer and metabolic diseases. In this review, we highlight recent advances in mitochondrial biogenesis from a metabolic perspective, with a particular emphasis on cancer. Metabolites act as donors for diverse chemical modifications, which have been systematically identified on mtDNA, rRNA, and tRNA. Besides, post-translational modifications of proteins involved in mtDNA replication, transcription, and translation has been revealed to connect metabolic signals with mitochondrial biogenesis. A comprehensive landscape of the mitochondrial central dogma has deepened our understanding of how mitochondria coordinate OXPHOS with other organelle-specific processes to obtain a flexible metabolic network, which potentiates tumor growth. Notably, non-canonical products and biological functions of the mitochondrial central dogma further reshape our concepts of cancer initiation and progression. Given that dysregulated mitochondrial biogenesis is found in multiple human disorders, including cancer, targeting this pathway offers new therapeutic opportunities. Genome-wide studies and drug screens have identified metabolic nodes and small molecules with potential to correct mitochondrial dysfunction in cancer, while emerging tools such as mtDNA editing enable precise intervention. Despite a maturing picture of mitochondrial biogenesis, many hidden players and functions remain to be uncovered to fully decipher mitochondrial biology in cancer.

Pharyngeal inflammation is a common upper respiratory tract disease characterized by increased inflammatory responses and mucin accumulation. Gentiopicroside (GPS), a natural compound with anti-inflammatory activity, has not been previously studied for its inhibitory effects on pharyngeal inflammation and its underlying mechanisms. This study aimed to investigate whether GPS inhibits pharyngeal inflammation in rats induced by Staphylococcus aureus (S. aureus) components and to elucidate the underlying mechanisms. In vivo, pharyngeal inflammation was induced in rats using S. aureus components, and GPS was administered to assess its effects on pharyngeal histopathological damage, mucosa injury, immune cell balance, secretory immunoglobulin A (SIgA) levels, cytokine production, and the expressions of E-cadherin, mucin5AC (MUC5AC), and cyclooxygenase-2 (COX-2). In vitro, human lung mucoepidermoid carcinoma cells (NCI-H292) were stimulated with lipoteichoic acid (LTA) to mimic S. aureus component-induced inflammatory stimulation. The effects of GPS on LTA-induced cytokine IL-1β and IL-6, COX-2, prostaglandin E2 (PGE2), MUC5AC, and E-cadherin expressions were evaluated. SiRNA against COX-2 was transfected, and a limited in vitro biochemical observation of GPS with recombinant MUC5AC was performed. GPS alleviated rat pharyngeal inflammation by improving pharyngeal histopathology, reducing mucosa injury, balancing immune cells, enhancing SIgA, and downregulating cytokines. It also inhibited the reduction of E-cadherin and the upregulation of MUC5AC and COX-2 in the rat pharynx and trachea. In NCI-H292 cells, GPS blocked LTA-induced increases in IL-1β, IL-6, COX-2, PGE2, and MUC5AC, as well as LTA-reduced E-cadherin. Transfection with siRNA against COX-2 further blocked LTA-induced MUC5AC expression, and GPS lost its inhibitory effect on MUC5AC expression, indicating that COX-2 was involved. Additionally, GPS showed a limited in vitro biochemical observation on recombinant MUC5AC, with uncertain physiological relevance. GPS alleviates S. aureus component-induced pharyngeal inflammatory damage in rats, with concomitant reductions in COX-2/PGE2 signaling activity and MUC5AC overexpression. These correlative observations provide a preliminary experimental basis for the potential application of GPS in ameliorating bacterial component-associated pharyngeal inflammation.

Activation of the STING pathway is essential for restoring immune surveillance against dormant disseminated tumor cells (DTCs) in the lungs. Inhaled Mn2+ has potential as a STING agonist; however, its clinical application is limited by the risk of chronic inflammation and metastasis, primarily due to reactive oxygen species (ROS) generation during inhalation. To address these risks, salvianolic acid B (salB) was identified as an effective ionophore for Mn2+, enhancing STING activation while mitigating ROS-induced inflammation. In this study, salB mitigated Mn2+-induced ROS levels and enhanced STING signaling, providing a safer, noninflammatory approach to activating immune surveillance in lung DTCs. The salB-Mn2+ complexes were encapsulated in human serum albumin nanoparticles (HSA NPs) for inhalation. PET and MRI analyses revealed that intratracheal administration of HSA NP@salB-Mn2+ restricted Mn2+'s systemic distribution, retaining it primarily in the lungs and minimizing central nervous system accumulation. Subsequent lung immunofluorescence further confirmed that HSA NP@salB-Mn2+ effectively targeted lung metastatic lesions. Despite this extended retention in lung tissue, histological analysis showed minimal inflammation in mice treated with HSA NP@salB-Mn2+, in contrast to those receiving MnCl2 or MnO. Consequently, HSA NP@salB-Mn2+ demonstrated superior suppression of 4T1 cell lung metastasis in postsurgical mice relative to MnCl2 or MnO. Mechanistically, salB functions as an agonist, independently activating p-STING, which synergizes with Mn2+-induced STING activation to significantly amplify signaling and downstream target engagement. In a postsurgical mouse model, the combination of HSA NP@salB-Mn2+ and αPD-1 antibody significantly reduced DTC dormancy and enhanced immune detection, confirming its immunotherapeutic potential. These findings establish salB as a promising inhalable ionophore for Mn2+ in DTC treatment, providing three key advantages: prolonged lung retention, reduced inflammation risk, and enhanced STING-activating efficacy.

BACKGROUND:Weichang'an pill (WCA) possesses potential advantages in promoting gastrointestinal motility and treating constipation. Ethanol extract (EE) and aqueous extract (AE) of WCA were used to investigate its efficacy in treating slow transit constipation (STC) and the material basis for exerting this effect. METHODS:The STC model was established in vivo by gavage of loperamide (Lop) in Sprague-Dawley rats, followed by gavage of WCA, EE, and AE. In vitro, norepinephrine (NE) was used to stimulate isolated ileal smooth muscle of rats to imitate the state of insufficient gastrointestinal motility during STC, and a model of excessive relaxation of isolated ileal smooth muscle was established. This model was used to observe and record the changes in contraction tension, amplitude, and frequency of ileal smooth muscle after treatment with WCA, EE, AE, and the active ingredients of WCA. KEY RESULTS:In vivo, WCA, EE, and AE treatment increased fecal parameters, improved gastrointestinal transit time, and alleviated pathological damage to the colon in STC rats. Its mechanism might be closely related to c-kit/SCF, RhoA/ROCK/MYPT1/MLC signaling pathways. In vitro, WCA, EE, AE, and the active ingredients of WCA, including costunolide (Cos), dehydrocostus lactone (Deh), agarotetrol (Aga), muscone (Mus), gallic acid (GA), oleic acid (Oleic), linoleic acid (Lin), umbelliferone (Umb), synephrine (Syn), ferulic acid (FA), chlorogenic acid (ChA), betaine (Bet), and riboflavin (Rib), significantly inhibited the NE-induced excessive relaxation of ileal smooth muscles. CONCLUSIONS:WCA, EE, and AE significantly improved constipation in STC rats. Moreover, the active ingredients in WCA, including Cos, Deh, Aga, Mus, GA, Oleic, Lin, Umb, Syn, FA, ChA, Bet, and Rib, might be the material basis for promoting intestinal motility.

The effective delivery of hydrophilic therapeutic agents (e.g., transition metal ions and protein antigens) remains a significant pharmacological hurdle in immunotherapy. Saponin-based delivery platforms demonstrate remarkable potential. Therapeutic modulation of the cGAS-STING pathway using manganese ions (Mn2+) holds significant promise for cancer immunotherapy. However, clinical translation faces additional safety challenges beyond delivery issues, since therapeutic doses of Mn2+ often induce chronic inflammation and oxidative stress. To address these challenges, we developed a biomimetic nanoparticle, Human serum albumin-Astragaloside IV-MnCl2 nanoparticles (HSA-A-M NPs), leveraging the dual functionality of the saponin astragaloside IV (AS-IV). AS-IV enhances Mn2+ uptake by increasing membrane permeability while suppressing NF-κB-driven inflammation and ROS production, improving safety. The nanoparticle's core, stabilized by HSA, boosts biocompatibility, while surface modifications with chitosan and hyaluronic acid (HA) derivatives (CS-NG@HGS) optimize tumor targeting, yielding the final formulation CS-NG@HGS@HSA-A-M. In vitro and vivo, this platform enhances cGAS-STING-mediated antitumor immunity while minimizing systemic Mn2+ toxicity and inflammation. By combining Mn2+ delivery with AS-IV's anti-inflammatory effects, it establishes a "controlled-activation" paradigm-potentiating IFN-I production yet curbing excessive immune responses. This dual-action design offers a safer, adaptable framework for metal ion-based combination therapies.

Immunosuppression increases disease risk, and the natural compound polydatin (PD) has been reported to modulate immune-related disorders. In cyclophosphamide-induced immunosuppressed mice, PD was evaluated for its immunomodulatory effects. Immune organ indices were measured, while H&E staining and ELISA assessed spleen pathology and serum cytokine levels. The proliferation of splenic lymphocytes, both total and subpopulation, was determined using concanavalin A or lipopolysaccharide stimulation, with flow cytometry analyzing peripheral blood and splenic lymphocytes, thymic T cell subtypes, cell cycling, and bromodeoxyuridine incorporation. Western blotting was used to assess Ki67, PCNA expression, and MAPK activation. PD significantly alleviated cyclophosphamide-induced reductions in spleen and thymus indices, improved the organization of red and white pulp in the spleen, and restored TNF-α and IFN-γ levels. It reversed cyclophosphamide-induced cell cycle arrest, characterized by increased PCNA and decreased Ki67, and corrected the diminished numbers of B and T cells and the reduced CD4+/CD8+ ratio in the thymus. In vitro, PD directly promoted splenic lymphocyte proliferation and cell cycling via MAPK activation. Overall, our findings demonstrated that PD alleviated mouse immunosuppression by activating splenic lymphocyte proliferation and re-organizing thymic T cell development and differentiation.

Background: Bladder outlet obstruction (BOO) is a prevalent urinary system disease main caused by benign prostatic hyperplasia (BPH) in males. Traditional Chinese medicine (TCM) Danzhi qing'e decoction (DZQE) has the functions of strengthening "Yang Qi" (one of the fundamental concept in TCM, representing the active, warming, and energizing force within the body. It governs physiological functions, maintains body temperature, and promotes vitality. Balanced "Yang Qi" supports immunity and metabolism, while deficiency may lead to fatigue, cold intolerance, or weakened resilience), promoting blood circulation, and removing blood stasis, and nourishing yin based on the TCM theory. Previous studies have found that it significantly improved BPH and regulate urinary function in estrogen and androgen-induced rats. However, it is unclear whether DZQE has an inhibitory effect on BOO rats. Methods: Male Wistar rats underwent retropubic partial bladder neck ligation to induce BOO. DZQE extract (2.7/5.4 g/kg) was administered orally for 35 days. Anesthetized rats underwent cystometry to assess BOO and treatment effects on urinary parameters. Bladder histopathology, fibrosis, and PCNA expression were evaluated via HE, Masson's, and IHC staining. Western blot quantified bladder tissue levels of choline acetyltransferase (ChAT), rho-associated protein kinase 1 (ROCK1), myosin light chain kinase (MLCK), myosin light chain 2 (MLC-2), extracellular signal-regulated kinase (ERK), proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2), and Bcl-2 related X protein (Bax). Primary bladder smooth muscle cells were cultured, exposed to hydrostatic pressure (HP, 3 h/24 h) using a custom apparatus to mimic BOO, and treated with DZQE components for Western blot analysis of ROCK1, MLCK, ERK, PCNA, and Bax. Results: The bladder hyperplasia, bladder index (BI) increased, and histopathological alteration were easily observed in BOO group, which were significantly inhibited by DZQE administration. DZQE also significantly inhibited the up-regulation of maximum voiding pressure (MVP) and down-regulation of residual urine volume (RV) observed in BOO rats. The expressions of ChAT and MLCK and the activation of ERK were much increased, while the expressions of ROCK1, MLC-2 and Bax were obviously decreased in BOO rats, all of which were then significantly inhibited by DZQE. In rat bladder smooth muscle cells (RBSMC), 3 h or 24 h duration of HP successfully simulated the BOO compensation and decompensation respectively in vitro. DZQE or its active components reduced the abnormal gene expressions in HP stimulated RBSMC. Conclusion: DZQE improves the urinary function in BOO rats mainly through the activation of ERK. Bakuchiol, salvianolic acid A, kaempferol, and tanshinone IIA are possibly the important active components of its therapeutic effects.

ObjectiveHuman immunodeficiency virus (HIV)-infected individuals who abuse methamphetamine (METH) exhibit more severe neurotoxicity and cognitive impairment. Pyroptosis, a programmed cell death pathway mediated by the inflammasome, has been implicated in various neurological diseases. This study aimed to elucidate the role of the AIM2 inflammasome in METH- and HIV-1 Tat-induced pyroptosis in human brain tissue and in vitro models.MethodsPostmortem brain tissue from HIV-infected individuals with a history of METH abuse was analyzed for pyroptosis markers and AIM2 inflammasome components using immunohistochemistry, immunofluorescence, and Western blotting. BV2 microglial cells were lentivirally transduced to knockdown AIM2 expression. DNA damage was assessed using Western blotting and the comet assay. Expression of pyroptosis-related proteins was evaluated by electron microscopy, Western blotting, and immunofluorescence. Cell viability was measured using the CCK8 assay.ResultsElevated levels of pyroptosis markers and AIM2 inflammasome components were observed in brain tissue from HIV-infected METH users. METH and Tat synergistically induced pyroptosis in BV2 cells in a time- and concentration-dependent manner, accompanied by DNA damage and activation of the AIM2 inflammasome. Knockdown of AIM2 significantly reduced the expression of pyroptosis-related proteins.ConclusionMETH and HIV-1 Tat proteins synergistically induce microglial pyroptosis by activating the AIM2 inflammasome through dsDNA damage. These findings suggest that targeting the AIM2 inflammasome may be a promising therapeutic strategy for HIV-associated neurocognitive disorder (HAND).

In the absence of tumor antigen specificity, direct chemokine administration carries the risk of significant “on-target, off-tumor” toxicities, highlighting the need for small-molecule approaches with reduced immunogenicity. This study investigates the synergistic potential of norcantharidin (NCTD) and lomitapide (lomi) in selectively restoring CCL4 expression by deactivating the tumor intrinsic β-catenin pathway. Due to its similar lipophilicity to lomi and potential to suppress β-catenin, NCTD prodrug (C12) was selected to be co-encapsulated with lomi in a nanoparticle-mediated co-delivery system (NP“C12 + lomi”). The NP“C12 + lomi” formulation exhibited a high encapsulation rate, uniform particle size, and suitability for therapeutic use. It effectively inhibited the proliferation of 4T1 cells and restored CCL4 expression. In both primary breast tumor and surgically resected tumor mouse models, NP“C12 + lomi” significantly increased the proportion of CD8+ cells in primary tumors, blood, and lung metastases, approximately doubling their presence. This led to a prolongation of median survival in mice to 59 days. Furthermore, when combined with an immune checkpoint inhibitor, NP“C12 + lomi” substantially inhibited tumor growth and lung metastasis without affecting body weight or causing major tissue or organ damage. This was attributed to the controlled dissociation of the nanoparticle and the subsequent modulation of C12 and lomi, which mitigated CCL4-related toxicity. This study provides valuable insights into the safe production of chemokines using a small-molecule pair through a nanosystem and presents a robust chemo-immunological cascade therapy strategy, demonstrating significant efficacy against malignant metastatic tumors.

Methamphetamine (METH) is currently considered one of the most notorious drugs globally. Chronic long-term METH abuse results in severe neurotoxicity, wherein oxidative stress and autophagy are key pathological phenomena and toxic phenotypes. However, the molecular mechanism by which METH induces oxidative stress and autophagy remains elusive. In this study, METH-induced autophagy and oxidative stress were replicated in both HT22 cells and C57BL/6 J mice. Notably, METH up-regulated the expression of chaperon protein sigma 1 receptor (S1R). However, METH-induced autophagy and oxidative stress were alleviated after targeted intervention with S1R using the chemical inhibitor, gene knockdown, or knockout techniques. More importantly, cannabidiol (CBD), a non-psychoactive natural cannabinoid derived from cannabis, exhibited therapeutic efficacy by down-regulating the high expression of S1R, autophagy, and oxidative stress following METH exposure both in vivo and in vitro. Overall, these results suggest that METH mediates autophagy and oxidative stress by up-regulating S1R expression, whereas CBD alleviates METH-induced autophagy and oxidative stress by suppressing S1R expression. This study expands our understanding of METH-induced neurotoxicity, identifying S1R as a potential therapeutic target against aberrant autophagy and oxidative stress, and further validates the medical value of CBD for the treatment of METH use disorder.

Cuscutae Semen (CS), a traditional herb recognized as a nutraceutical food in China, has been widely utilized in managing aging-related diseases throughout history. However, whether this mechanism is associated with mitochondrial stress tolerance remains unclear. In the present study, Caenorhabditis elegans (C. elegans) was used to investigate the effects of CS on their longevity. The data demonstrated that CS prolonged the average lifespan of the nematodes by 15.26%, reducing lipofuscin accumulation by 61.46%, as well as improving spontaneous motility. CS treatment significantly enhanced the resistance of C. elegans to hydrogen peroxide-induced oxidative stress and 37 °C induced heat stress, reducing reactive oxygen species (ROS) production by 71.45%. Additionally, membrane potential (MMP) and adenosine triphosphate (ATP) were increased by 354.72% and 69.64%, respectively. However, mitochondrion-specific ROS and calcium flux were significantly reduced to 45.86% and 63.25%, respectively, in C. elegans treated with CS. Consistently, the polymerase chain reaction data revealed that CS significantly up-regulated the expressions of the antioxidant-related genes skn-1, ctl-1, sod-3, and gst-4; the heat shock gene hsp-16.2; and the autophagy-related genes lgg-1 and bec-1. Considering the crucial role of the silent information regulator sirtuin 1 (SIR-2.1/SIRT1) in aging-related mitochondrial oxidative stress, we examined its expression and transcriptional activity. As expected, treatment with CS induced SIRT1 expression, and isorhamnetin identified from CS extract significantly enhanced SIRT1 transcriptional activity in HEK293T cells. Collectively, our results provided evidence that CS prolonged the lifespan of C. elegans by ameliorating oxidative stress damage and mitochondrial dysfunction via SIRT1.

Traditional alkaloid extraction processes (AEP) usually utilize large amount of organic reagents or acidic solvents, which produces considerable toxic/acidic solid and liquid waste. In order to make AEP cleaner, we take the extraction process of Sophora Flavescens and Heterosmilax Japonica as an instance and introduce process analytical technology (PAT) to analyze and improve the process. Specifically, we use near infrared spectroscopy (NIRS) to collect the process information. By combining NIRS data with online sampling and off-line high-performance liquid chromatography (HPLC) measurements, we find the hot leaching process (HLP) of the traditional AEP is quite inefficient and the extraction is quite incomplete. Hence, we make corresponding modifications to the process, including dropping the HLP step and introducing a three-phase static leaching step. Furthermore, we build predictive models using the NIRS and HPLC data in order to offer timely predictions of critical quality attributes (CQA), which makes it possible to perform dynamic controlling actions. With the help of PAT, we finally achieve flexible AEP which is cleaner and has higher extraction rate (similar to 65% more) and lower solvent consumption (similar to 23% less) than the traditional AEP. The flexible AEP also has much shorter processing time (similar to 60% less), which can lead to better energy consumption.

Necroptosis is a programmed form of necrosis, and compounds inducing necroptosis may contribute to cancer treatment. 20(S)-ginsenoside Rg3 is a natural compound extracted from ginseng, which exhibited a broad-spectrum of antitumor activity. In the present study, the potential role of 20(S)-ginsenoside Rg3 in inducing necroptosis in prostate cancer cells was evaluated. 20(S)-ginsenoside Rg3 inhibited the proliferation of prostate cancer cells and upregulated the expression of necroptotic proteins such as receptor-interacting serine/threonine-protein kinase 1 (RIPK1), RIPK3, and their downstream mixed lineage kinase domain-like protein (MLKL). Pretreatment with the selective RIPK1 inhibitor necrostatin-1 (Nec-1) partially reversed the inhibitory effect of 20(S)-ginsenoside Rg3 on prostate cancer cell proliferation. 20(S)-ginsenoside Rg3 led to the accumulation of reactive oxygen species (ROS) and the regulation of autophagy in cancer cells. Scavenging ROS with N-acetyl-L-cysteine (NAC) antagonized the regulatory effects of 20(S)-ginsenoside Rg3 on cell autophagy and necroptotic proteins expression. Moreover, 20(S)-ginsenoside Rg3 exhibited an antitumor effect in a prostate cancer xenograft mouse model in which it upregulated the expression of RIPK1, RIPK3, MLKL and led to a decrease in tumor weight, as well as an increase in necrotic areas in tumor tissue. In conclusion, our study showed that 20(S)-ginsenoside Rg3 might induce necroptosis in prostate cancer in vitro and in vivo via the ROS/autophagy signaling pathway.