This study investigated the mechanism by which notoginsenoside R1 (NGR1) alleviates neuroinflammation by regulating metabolic disorders in microglia through network pharmacology, in vitro experiments, and untargeted metabolomics. Neuroinflammation-related disease targets and NGR1 targets were obtained through multi-database mining, overlapping targets were screened using Venny, and GO and KEGG enrichment analyses as well as protein-protein interaction network construction were performed to screen core targets. Molecular docking software was used to evaluate the binding ability between NGR1 and the core targets. An inflammatory model was established by lipopolysaccharide-induced BV2 microglial cells, and Western blot was used to detect the effects of different concentrations of NGR1 on the expression of inflammation-related proteins in microglia, and to determine the effective concentrations of NGR1 affecting the NF-κB signaling pathway. Untargeted metabolomic analysis was conducted within the effective concentration range of NGR1, and differential metabolites were screened by combining multivariate statistical analysis with metabolic pathway enrichment analysis. Cytoscape and MetaboAnalyst platforms were further used to screen key metabolites closely associated with the NF-κB signaling pathway. Exogenous metabolite supplementation experiments were conducted to verify whether NGR1 could inhibit the activation of the NF-κB signaling pathway by regulating the levels of related metabolites. The results showed that there were 43 overlapping genes between neuroinflammation-related disease targets and NGR1 targets, among which 16 were core targets. KEGG enrichment results indicated that the overlapping targets were significantly enriched in inflammatory signaling pathways such as NF-κB. Molecular docking results showed that NGR1 had high binding affinity with multiple NF-κB signaling pathway proteins. Western blot results demonstrated that NGR1 could significantly regulate the expression of NF-κB signaling pathway-related proteins and inflammatory factors, and 25 and 50 μmol·L^-1 were the effective concentrations affecting the NF-κB signaling pathway. Metabolomic results showed that NGR1 could significantly affect multiple pathways such as glutathione metabolism. Metabolite-target association analysis results showed that NGR1 could reverse abnormal changes in metabolites closely related to NF-κB signaling pathway targets, including homocysteine, L-leucine, and prostaglandin E2 ethanolamide. Exogenous metabolite supplementation experiments showed that homocysteine could weaken the inhibitory effect of NGR1 on the NF-κB signaling pathway, suggesting that homocysteine may be involved in the NGR1-mediated NF-κB inhibitory process. In summary, NGR1 may inhibit the activation of the NF-κB signaling pathway through both direct inhibition and indirect regulation of homocysteine metabolism, thereby alleviating neuroinflammatory responses.

Tumor-associated macrophages (TAMs) have a dual role in the tumor microenvironment, mediating both immune activation and suppression. M1-TAMs kill tumor cells, while M2-TAMs are immunosuppressive and promote tumor growth. The activation of TAMs and enhancement of their anti-tumor immune function is a major goal in cancer immunotherapy. Here, we report a novel pyrazole derivative, IHMT-140, which reprograms tumor-promoting M2 macrophages into tumor-suppressing M1 macrophages, thereby inhibiting tumor growth. Mechanistically, IHMT-140 raises lysosomal pH in M2 macrophages, blocking autophagy and activating MAPK and NF-κB signaling, which induces M2-TAMs to M1-TAMs and mediates the anti-tumor response. As a potential immunomodulator, IHMT-140 provides not only a new tool for studying TAM reprogramming, but also a foundation for developing related immunotherapies. All animal experimental protocols were approved by the Hefei Institutes of Physical Science, Chinese Academy of Sciences, with approval number DWLL-2023-116.

Based on the cellular proliferation tracing technique (ProTracer), this study explored the role of rifampicin (RIF) in promoting liver regeneration in a mouse model with zone-specific liver injury. The ProTracer system enables in vivo recording of cell proliferation by labeling proliferating cells with GFP fluorescence. Carbon tetrachloride (CCl₄)-induced injury and bile duct ligation (BDL) models were established, followed by RIF or vehicle treatment. Liver function was assessed by measuring serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bile acids (TBA), total bilirubin (TBIL), and direct bilirubin (DBIL). Liver injury area was evaluated by H&E staining, while whole-tissue fluorescence imaging was used to assess liver regeneration. Immunofluorescence co-staining was performed to quantify proliferating hepatocytes, identify cell types, and analyze zonal distribution. RNA sequencing was conducted to identify key differentially expressed genes and signaling pathways, with Western blot used for protein-level validation. In vitro validation was performed using the HepG2 hepatocyte cell line. Results showed that RIF significantly reduced serum ALT and AST levels in CCl₄-induced liver injury and decreased TBA, TBIL, and DBIL levels in the BDL-induced liver injury, effectively reducing the area of liver damage. Whole-tissue fluorescence imaging revealed enhanced GFP signals and increased proliferating cells in the RIF-treated group. Immunofluorescence analysis indicated that RIF primarily promoted hepatocyte proliferation, with a marked increase in proliferating cells in zone 2 of the liver lobule (approximately 2.5-fold), and also significant increased in zones 1 and 3. RNA-Seq and Western blot analyses demonstrated upregulation of cell cycle and proliferation-related genes such as the Mcm family, Ccnd1, Ccna2, and Mki67, with enrichment in the cell cycle pathway. In vitro experiments showed that RIF treatment enhanced HepG2 cell viability and increased the proportion of Ki67-positive cells. In conclusion, RIF promotes hepatocyte proliferation and enhances liver regeneration following zone-specific liver injury by activating cell cycle signaling pathways. Animal experiments were conducted with approval from the Animal Ethics Committee of Binzhou Medical University (approval number: 2025-2018).

This study aimed to construct a stable cell line of human embryonic kidney epithelial cells 293 (HEK293) expressing recombinant human coagulation factor Ⅸ (rhFⅨ) and to identify the procoagulant biological activity of the expressed product. The recombinant plasmid pcDNA3.1-FⅨ was transfected into HEK293 cells, and a monoclonal stable transfection cell line was obtained through geneticin (G418) pressure screening. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine whether the FⅨ gene was transcribed and expressed in the monoclonal stable transfection cell line. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot were used to identify the expression of rhFⅨ; the concentration of rhFⅨ was determined by bicinchoninic acid assay (BCA), and the procoagulant activity of rhFⅨ was determined by activated partial thromboplastin time (APTT). The results showed that after G418 (200 µg·mL^-1) pressure screening, 5 monoclonal stable cell lines could be normally transcribed, and the protein expression could be achieved through the transcription-translation process; the protein in the cell supernatant was purified and showed a single band of rhFⅨ; the expression level of rhFⅨ reached (1.58 ± 0.10) mg·L^-1, and the maximum procoagulant activity was (66.8 ± 0.5)%. The established HEK293 monoclonal stable transfection cell line can efficiently and stably express rhFⅨ and has good procoagulant activity, providing a cell source for promoting the pharmacological evaluation and preclinical research of rhFⅨ.

This study was to investigate the protective effect of Saposhnikovia divaricata (Turcz.) Schischk. (SD) on ulcerative colitis (UC) and its relationship with aryl hydrocarbon receptor (AHR)/NOD-like receptor family, pyrin domain containing 3 (NLRP3) pathway. The acute colitis model of C57BL/6J mice was established by continuous drinking water with 2.5% dextran sulfate sodium (DSS) for 7 days. At the same time, the ethanol extract of Saposhnikovia divaricata (2 and 5 g·kg^-1·d^-1) or 5-aminosalicylic acid (5-ASA, 100 mg·kg^-1·d^-1) was given by gavage. Body weight and disease activity index (DAI) were recorded. Colon length, hematoxylin-eosin (HE) and Alcian blue-periodic acid-Schiff staining (AB-PAS) staining and immunofluorescence were used to evaluate colon tissue damage and changes of goblet cells and tight junction proteins. ELISA, RT-qPCR and Western blot were used to analyze the expression of inflammatory factors, tight junction proteins and AHR/NLRP3-related molecules in serum and colon tissues. The use of animals in this study was approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine (approval number: PZSHUTCM2510310022). The results showed that compared with the DSS model group, Saposhnikovia divaricata could dose-dependently reduce DSS-induced weight loss and DAI increase, prolong colon length, and reduce mucosal pathological damage. At the same time, it down-regulated the expression of inflammatory factors such as TNF-α, IL-1β and IL-6 in serum and colon tissues, up-regulated the mRNA and protein levels of zonula occludens-1 (ZO-1) and occludin tight junction proteins, and increased the expression of AHR protein, and inhibited NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1 and IL-1β. In AHR knockout mice, the alleviation effect of the ethanol extract of Saposhnikovia divaricata on DSS-induced acute ulcerative colitis in mice was not observed. In summary, Saposhnikovia divaricata may inhibit the activation of downstream NLRP3 inflammasome by activating AHR, thereby maintaining intestinal mucosal barrier function and reducing intestinal inflammation, providing experimental basis for its application in the prevention and treatment of UC.

Alzheimer's disease (AD) is a chronic neurodegenerative disorder. Current medications can only alleviate symptoms but have not yet achieved a cure. Given its multifactorial pathogenesis, the development of small-molecule compounds that act simultaneously on multiple targets is particularly important. Traditional Chinese medicine (TCM) small molecules, which can regulate multiple biological targets and signaling pathways, offer unique advantages in AD treatment. This study utilized genipin, a TCM small molecule with potential neuroprotective effects, as the lead compound to design and synthesize a series of multi-target anti-AD small molecule compounds. Among these, compounds JD-5 and JD-7 exhibited significantly enhanced GSK-3β inhibitory activity (JD-5, IC₅₀ = 3.54 μmol·L^-1; JD-7, IC₅₀ = 2.47 μmol·L^-1), substantially outperforming the lead compound genipin (IC₅₀ = 28.16 μmol·L^-1). Furthermore, both JD-5 and JD-7 demonstrated significant neuroprotective effects in SH-SY5Y cell damage experiments induced by tBHP and Aβ_25-35. Further investigation revealed that compounds JD-5 and JD-7 could significantly reduce Tau phosphorylation (S396) levels in primary neuronal cells, with dose-dependent effects. All animal experiments were approved by the Experimental Animal Ethics Committee of Nanjing Ramda Pharmaceutical Co., Ltd (approval number: IACUC-20220202). Based on these results, it is hypothesized that compound JD-5 likely exerts neuroprotective and antioxidant stress effects by inhibiting GSK-3β activity and reducing Tau protein phosphorylation levels, providing a reference for the development of multifunctional anti-AD drugs.

This study employs silica gel column chromatography and thin-layer chromatography to study the chemical constituents of the effective substance groups from Artemisia ordosica, and analyze the spectral data of the separated compounds to determine their structures. Nine alkynes, including ordosin (1), capillene (2), capillin (3), capillinol (4), trans-dehydromatricaria ester (5), capillarin (6), o-hydroxycapillene (7), capillaridin (8), o-hydroxycapillin (9), and one dihydroflavone, 5-hydroxy-7,4′-dimethoxydihydroflavone (10), were isolated and identified from the effective substance groups of Artemisia ordosica. Among these, compound 1 is a new alkyne, while 2-4, 6, 8, and 9 were isolated for the first time from Artemisia ordosica. By examining the effects of compounds 1-4, 6, 8, and 9 on the release of nitric oxide (NO) in the culture medium of RAW264.7 macrophages induced by lipopolysaccharide (LPS), the inhibitory effect on inflammatory response was evaluated. The results revealed that compounds 1-4, 6, 8, and 9 exhibited NO inhibitory activity at a concentration of 10 μmol·L^-1, with inhibition rates of 96.7%, 88.4%, 91.6%, 77.8%, 98.7%, 86.7% and 75.1%, respectively.

The ethyl acetate fraction of the 95% ethanol extract from black mulberry branches was isolated and purified by comprehensive application of chromatographic techniques, including normal-phase silica gel column chromatography, Sephadex LH-20 gel column chromatography, and semi-preparative high-performance liquid chromatography (semi-prep HPLC). Fourteen compounds were obtained, and their structures were identified using modern spectroscopic techniques (UV, IR, MS, NMR). These compounds were identified as nigrasin M (1), nigrasin N (2), sanggenol F (3), sanggenon A (4), nigrasin K (5), nigrasin H (6), petalopurpurenol (7), solophenol D (8), nigragenon A (9), nigragenon I (10), nigragenon J (11), notabilisin A (12), notabilisin C (13), and licoflavone C (14). Among them, compounds 1 and 2 are new flavonoids, while compounds 13 and 14 were isolated from Morus nigra branches for the first time. To evaluate the hypoglycemic potential of compounds 1-13, a glucose uptake assay was performed using an L6 cell model. The results showed that compounds 6, 9, 11, and 12 significantly enhanced glucose uptake in L6 cells, suggesting their potential hypoglycemic activity.

Nine sesquiterpenoids were isolated from the dichloromethane fraction of the 95% ethanol extract from the roots of Lindera glauca by using a combination of separation techniques, including silica gel, ODS and Sephadex LH-20 column chromatographies, as well as semi-preparative HPLC. Based on a comprehensive analysis of modern spectroscopic data, such as IR, UV, NMR, HRESIMS and ECD, the structures of isolated compounds were identified as glaucatotone M (1), glaucatotone K (2), glaucatotone G (3), pseudoguaianelactone C (4), pseudoguaianelactone B (5), elema-1,3,11(13)-trien-12-oic acid (6), α-costic acid (7), 3β-hydroxycostic acid (8) and rupestonic acid I (9), respectively. Compound 1 was a new guaiane-type sesquiterpenoid, and compounds 6 and 7 were first reported from the genus Lindera. The inhibitory activity of compounds 1-7 and 9 against both the proliferation of TNF-α-induced rheumatoid arthritis fibroblast-like synoviocytes cell line MH7A and the release of pro-inflammatory cytokines (IL-6 and IL-1β) were evaluated. The results showed that compound 1 had a significant inhibitory effect against TNF-α-induced MH7A cell proliferation and reduced the production of IL-6 and IL-1β effectively.

The 95% ethanol extract of Curcuma longa was subjected to isolation and purification using silica gel column chromatography, C₁₈ column chromatography, and semi-preparative HPLC. The planar structures of the isolated compounds were elucidated by HR-MS, IR, and NMR spectroscopy. The absolute configurations of the new compounds were determined by calculated electronic circular dichroism (ECD) and DP4+ NMR analysis. Finally, three bisabolane-type sesquiterpenoids were isolated and identified from C. longa as (1R,7S)-15-norbisabolane-2-ene-11-ol-4,9-dione (1), (7S)-15-norbisabolane-1,3,5,10-tetraene-4-ol (2), and 8-hydroxyl-ar-turmerone (3). Compounds 1 and 2 were two novel 15-norbisabolane-type sesquiterpenoids.