The widespread use of aspartame has raised increasing health concerns, particularly regarding its potential endocrine-disrupting effects and a possible association with polycystic ovary syndrome (PCOS), a condition marked by endocrine and metabolic abnormalities. This study systematically investigated the molecular mechanisms by which aspartame may contribute to PCOS pathogenesis using a network toxicology approach. A total of 281 common targets between aspartame and PCOS were identified. Protein-protein interaction network analysis and five topological algorithms revealed 20 hub targets significantly enriched in pathways such as endocrine resistance, TNF signaling, and progesterone-mediated oocyte maturation. Eight of these targets showed significant differential expression in PCOS patients. Weighted gene co-expression network analysis (WGCNA) further demonstrated that several of these hub targets are embedded in co-expression modules positively associated with PCOS phenotypes. Transcription factor prediction indicated that these targets are primarily regulated by FOS and JUN. Molecular docking showed favorable binding affinities between aspartame and both transcription factors. Subsequent molecular dynamics simulations supported the dynamic stability of aspartame binding to FOS and JUN. These findings suggest that aspartame may contribute to PCOS by modulating FOS- and JUN-mediated transcriptional programs, thereby disrupting biological pathways related to endocrine regulation and inflammation. Collectively, this study reveals a potential mechanism by which aspartame mediates PCOS and offers novel insights for toxicological assessment of food additives.

Rationale:Inflammatory bowel disease (IBD), known for its complexity and frequent relapses, urgently demands novel therapeutics due to the limited efficacy of current treatments. Cinnamaldehyde (CMA), a bioactive compound derived from Cinnamomum cassia Presl, has exhibited therapeutic potential for IBD. However, the therapeutic mechanism of CMA remains incompletely elucidated, and clinical translation is hampered by its poor oral pharmacokinetics. Methods:Using RAW 264.7 cells stimulated with either LPS or IL-4, we evaluated the effects of CMA on macrophage polarization. Subsequently, the impact of CMA on glucose metabolism in M1 macrophages was analyzed. RNA sequencing identified the signaling pathways through which CMA inhibits M1 macrophage polarization, and this was further validated through genetic or pharmacological blockade. To overcome the pharmacokinetic challenges of CMA, macrophage membrane-biomimetic CMA-loaded nanoparticles (MM@CMANP) were designed, and their pharmacokinetics and targeting to intestinal inflammation sites were evaluated. Finally, the efficacy of MM@CMANP was assessed in DSS-induced IBD mice. Results:CMA suppresses M1 macrophage polarization in vitro. Notably, CMA disrupted M1 macrophage glucose metabolic reprogramming, characterized by glycolysis suppression and enhanced oxidative phosphorylation. RNA sequencing demonstrated a clear association with mitophagy pathway following CMA treatment, and mechanistic studies verified that CMA promotes BCL2/adenovirus E1B 19 kDa-interacting protein 3 (BNIP3)-mediated mitophagy activation. Crucially, CMA-induced inhibition of M1 macrophages was mitigated by BNIP3 knockdown or autophagy inhibitors. MM@CMANP enhanced CMA accumulation in inflamed colonic tissues. In IBD mice, MM@CMANP significantly alleviated epithelial barrier disruption and mucosal inflammation. Consistent with in vitro findings, CMA modulated macrophage polarization and autophagy in vivo. Conclusions:These results establish mitophagy as a central mechanism underlying anti-IBD effects of CMA and position MM@CMANP as a clinically translatable nanotherapeutic platform for IBD.

Inflammation is an important predisposing factor for many chronic diseases. The dietary flavonoid silibinin (SB) has excellent anti-inflammatory properties in cells, but its low bioavailability in the blood compromises its therapeutic potential. This study aims to investigate the potential of dibenzoylmethane (DBM) to synergistically enhance the anti-inflammatory benefits of SB. The synergistic effects of DBM and SB in combination were evaluated in lipopolysaccharide (LPS)-induced RAW264.7 cells and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced mice. In addition, a network pharmacology approach and molecular docking were used to explore the key targets and signaling pathways of DBM and SB in combination. The results showed that DBM and SB synergistically inhibited the production of nitric oxide (NO), reactive oxygen species (ROS), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in a 1:1 concentration ratio. These two compounds may exert their synergistic effects by modulating the nuclear factor kappa-B (NF-κB) and HIF-1 signaling pathways, among others. Molecular docking revealed that both compounds exhibited high binding affinities to inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Compared with single-compound use, the two compounds in combination significantly reduced ear edema and inflammatory cell infiltration and inhibited the protein expression of iNOS and COX-2 in TPA-induced mice. This research provides a rationale for the combination of DBM and SB as an effective anti-inflammatory agent.

Background and objective: Patients with rheumatoid arthritis (RA) are more susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) than healthy population, but there is still no therapeutic strategy available for RA patients with corona virus disease 2019 (COVID-19). Guizhi-Shaoyao- Zhimu decoction (GSZD), Chinese ancient experience decoction, has a significant effect on the treatment of Rheumatism and gout. To prevent RA patients with mild-to-moderate COVID-19 from developing into severe COVID-19, this study explored the potential possibility and mechanism of GSZD in the treatment of this population. Methods: In this study, we used bioinformatic approaches to explore common pharmacological targets and signaling pathways between RA and mild-to-moderate COVID-19, and to assess the potential mecha- nisms of in the treatment of patients with both diseases. Beside, molecular docking was used to explore the molecular interactions between GSZD and SARS-CoV-2 related proteins. Results: Results showed that 1183 common targets were found in mild-to-moderate COVID-19 and RA, of which TNF was the most critical target. The crosstalk signaling pathways of the two diseases focused on innate immunity and T cells pathways. In addition, GSZD intervened in RA and mild-to-moderate COVID-19 mainly by regulating inflammation-related signaling pathways and oxidative stress. Twenty hub compounds in GSZD exhibited good binding potential to SARS-CoV-2 spike (S) protein, 3C-like protease (3CLpro), RNA-dependent RNA polymerase (RdRp), papain-like protease (PLpro) and human angiotensin- converting enzyme 2 (ACE2), thereby intervening in viral infection, replication and transcription. Conclusions: This finding provides a therapeutic option for RA patients against mild-to-moderate COVID- 19, but further clinical validation is still needed.

Anti-tumor activity of Tremella fuciformis polysaccharides (TFPS) has been widely reported, but its mechanism remains poorly understood. In this study, we established an in vitro co-culture system (B16 melanoma cells and RAW 264.7 macrophage-like cells) to explore the potential anti-tumor mechanism of TFPS. Based on our results, TFPS exhibited no inhibition on the cell viability of B16 cells. However, significant apoptosis was observed when B16 cells were co-cultured with TFPS-treated RAW 264.7 cells. We further found that mRNA levels of M1 macrophage markers including iNOS and CD80 were significantly upregulated in TFPS-treated RAW 264.7 cells, while M2 macrophage markers such as Arg-1 and CD 206 remained unchanged. Besides, the migration, phagocytosis, production of inflammatory mediators (NO, IL-6 and TNF-α), and protein expression of iNOS and COX-2 were markedly enhanced in TFPS-treated RAW 264.7 cells. Network pharmacology analysis indicated that MAPK and NF-κB signaling pathways may be involved in M1 polarization of macrophages, and this hypothesis was verified by Western blot. In conclusion, our research demonstrated that TFPS induced apoptosis of melanoma cells by promoting M1 polarization of macrophages, and suggested TFPS may be applied as an immunomodulatory for cancer therapy.

BACKGROUND AND OBJECTIVE:Patients with rheumatoid arthritis (RA) are more susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) than healthy population, but there is still no therapeutic strategy available for RA patients with corona virus disease 2019 (COVID-19). Guizhi-Shaoyao-Zhimu decoction (GSZD), Chinese ancient experience decoction, has a significant effect on the treatment of Rheumatism and gout. To prevent RA patients with mild-to-moderate COVID-19 from developing into severe COVID-19, this study explored the potential possibility and mechanism of GSZD in the treatment of this population. METHODS:In this study, we used bioinformatic approaches to explore common pharmacological targets and signaling pathways between RA and mild-to-moderate COVID-19, and to assess the potential mechanisms of in the treatment of patients with both diseases. Beside, molecular docking was used to explore the molecular interactions between GSZD and SARS-CoV-2 related proteins. RESULTS:Results showed that 1183 common targets were found in mild-to-moderate COVID-19 and RA, of which TNF was the most critical target. The crosstalk signaling pathways of the two diseases focused on innate immunity and T cells pathways. In addition, GSZD intervened in RA and mild-to-moderate COVID-19 mainly by regulating inflammation-related signaling pathways and oxidative stress. Twenty hub compounds in GSZD exhibited good binding potential to SARS-CoV-2 spike (S) protein, 3C-like protease (3CLpro), RNA-dependent RNA polymerase (RdRp), papain-like protease (PLpro) and human angiotensin-converting enzyme 2 (ACE2), thereby intervening in viral infection, replication and transcription. CONCLUSIONS:This finding provides a therapeutic option for RA patients against mild-to-moderate COVID-19, but further clinical validation is still needed.

In China, Curcuma plants have been widely used as traditional Chinese medicine in the treatment of inflammation-related diseases for thousands of years. This study aimed to investigate the anti-neuroinflammatory effect of essential oils (EOs) from four Curcuma species, including Curcuma longa L. (CL), Curcuma kwangsiensis S.G. Lee and C.F. Liang (CK), Curcuma zedoaria (Christm.) Roscoe (CZ) and Curcuma aromatica Salisb. (CA), and to explore the underlying mechanisms using metabolomics and network pharmacology. A total of 49 chemical compositions were identified in EOs from four Curcuma species, and the main compounds included ar-turmerone (67.76%), curzerenone (2.33–50.43%), nootkatone (14.41%), curlone (12.93%), β-Elemene (2.15–11.47%), curzerene (1.55–11.01%), 1,8-Cineole (11.00%) and humulene (1.89–10.99%). In the Morris water maze test, these EOs significantly improved cognitive impairment in LPS-induced mice. Treatment of these EOs protected neurons and inhibited microglia activation. Besides, a significant reduction of inflammatory cytokines (IL-6, TNF-α, IL-1β and MCP-1) was detected in the brain tissue of mice. Based on the metabolomics of blood, 11 metabolites mainly associated with arachidonic acid metabolism, linoleic acid metabolism, and aminoacyl-tRNA biosynthesis were identified. Network pharmacology further validated these metabolic pathways and suggested that EOs may interfere with multiple neurodegenerative diseases and inflammation-related signaling pathways by modulating metabolic changes. In conclusion, this study demonstrates the inhibitory effect of four Curcuma EOs on neuroinflammation and suggests that they may be candidates for the treatment of neuroinflammation.

The fluorescence properties of hemocyanin from the scorpion Leirus quinquestriatus were studied. Emission and excitation spectra were determined for protein in both its oxygenated and deoxygenated forms. Oxygenation was found to bring about a large blue shift in the position of the fluorescence maximum, in addition to a marked quenching of the fluorescence intensity as noted before for another hemocyanin. An appreciable tyrosyl contribution to the fluorscence of oxyhemocyanin was inferred from the wavelength dependence of its emission and excitation spectra. No tyrosyl fluorescence could be observed in either apo- or deoxyhemocyanin. It was concluded that the inability to observe tyrosyl emmision in deoxyhemocyanin is due to the dominating emission to tryptophan. The implication of the findings to the often noted failure to detect tyrosyl emission in proteins containing both tyrosine and tyrptophan is discussed.

Climate change has an extremely important impact on the geographic distribution of plants. The genus Millettia is an important plant resource in China and is widely used in medicine and ornamental industries. Due to the continuous changes of climate and the development and utilization of plant resources of the genus Millettia, it is of great significance to systematically investigate the geographic distribution of plants of the Millettia and their potential distribution under climate change. DIVA-GIS software was used to analyze 3492 plant specimens of 35 species of genus Millettia in the herbarium, and the ecological geographic distribution and richness of Millettia were analyzed, and the MaxEnt model was used to analyze the current and potential distribution in the future. The results show that the genus Millettia is distributed in 30 provinces in China, among which Yunnan and Guangdong provinces are the most distributed. Our model determines that precipitation in the driest month and annual temperature range are the most important bioclimatic variables. Future climate changes will increase the suitable habitat area of M. congestiflora by 16.75

Zingiber striolatum Diels (Z. striolatum), a widely popular vegetable in China, is famous for its medicinal and nutritional values. However, the anti-inflammatory effects of essential oil from Z. striolatum (EOZS) remain unclear. In this study, EOZS from seven regions in China were extracted and analyzed by GC–MS. LPS-induced RAW264.7 cells and 12-O-Tetradecanoylphorbol 13-acetate (TPA)-stimulated mice were used to evaluate the anti-inflammatory effects of EOZS. Results show that 116 compounds were identified in EOZS from seven locations. Samples 2, 4 and 5 showed the best capability on DPPH radical scavenging and NO inhibition. They also significantly reduced the production of ROS, pro-inflammatory cytokines, macrophage morphological changes, migration and phagocytic capability. Transcriptomics revealed MAPK and NF-κB signaling pathways may be involved in the anti-inflammatory mechanism, and the predictions were proven by Western blotting. In TPA-induced mice, EOZS reduced the degree of ear swelling and local immune cell infiltration by blocking the activation of MAPK and NF-κB signaling pathways, which was consistent with the in vitro experimental results. Our research unveils the antioxidant capability and potential molecular mechanism of EOZS in regulating inflammatory response, and suggests the application of EOZS as a natural antioxidant and anti-inflammatory agent in the pharmaceutical and functional food industries.

Background/aim: Naringin, a naturally occurring flavanone glycoside, can inhibit oxidative stress and inflammatory reactions. Atorvastatin, which belongs to the class of drugs called statins, is an inhibitor of 3-hydroxyl-3-methylglutaryl coenzyme A (HMG-CoA) reductase and it has shown anticancer activity in prostate cancer (PCa). The present study investigated whether the combination of atorvastatin and naringin has an effect on inhibiting growth in PCa and the mechanisms underlying this effect. Methods: Cell growth was assessed by CCK-8 and trypan blue exclusion assay, and the IC50 of LNCaP and PC-3 cells were determined by MTT assay. Scratch migration and matrigel-coated transwell invasion were used to assess both migration and invasion in prostate cancer cell lines. The expression levels of p-Akt, p-STAT3, survivin, AR, Bcl-2, and Bax in prostate cancer cell lines were determined by Western blot. PC-3 and LNCaP xenografts were applied to assess the effect in vivo. Results: Naringin in combination with atorvastatin had a synergistic inhibitory effect on growth in the prostate cancer cell lines PC-3 and LNCaP, and the combination more strongly inhibited migration and invasion than either single drug. The combination of atorvastatin with naringin potently inhibited the expression levels of AR, p-Akt, survivin, p-STAT3, and Bcl-2. However, an increase in the level of p-STAT3 was found in the PC-3 cells treated with atorvastatin and naringin alone, which differed from that observed in LNCaP. Moreover, the level of Bax in the prostate cancer cell lines treated with atorvastatin and naringin was higher than in those treated with either drug alone. Atorvastatin or naringin treatment had an inhibitory effect on the growth of PC-3 and LNCaP tumors in vivo, and this was especially true of the combined treatment. Conclusions: The data suggest that the combination of atorvastatin and naringin may be an effective method for inhibiting the growth of prostate cancer. Further research is needed to determine the mechanism of action of the combination of atorvastatin and naringin.

Climate change has a crucial impact on plant resources distribution, especially on agricultural crops. Species belonging to genus Zingiber are important crop resources in China, and their rhizomes are widely used in medicine and food. In this study, the ecological distribution and richness of the genus Zingiber were analyzed using a DIVA-GIS analysis of 967 georeferenced herbarium records to produce present and future distribution simulations using the MaxEnt model. The results showed that the genus Zingiber is widely distributed in 28 provinces in China, with the greatest richness in Guangxi, Guangdong, and Yunnan. Based on our model, precipitation of the driest month, precipitation of the warmest quarter, and the minimum temperature of the coldest month were the most significant bioclimatic variables controlling the distribution of five selected Zingiber species. Future climate change will likely result in the increase of suitable habitat area for Zingiber officinale by 2.85%, but dramatically decrease that for Zingiber striolatum by 18.04%, Zingiber zerumbet by 17.87%, Zingiber corallinum by 12.40%, and Zingiber mioga by 9.46%. Thus, conservation measures should be taken to preserve these valuable resources.

Eriocitrin is a flavonoid that is isolated from orange peel. Resveratrol is a polyphenol compound, which is present in various fruits, such as grapes. The aim of the present study was to investigate both in vitro and in vivo anti-inflammatory effects of eriocitrin combined with resveratrol using lipopolysaccharide (LPS)-induced RAW264.7 cells and a mouse model of ear edema. The results showed that eriocitrin combined with resveratrol strongly inhibited LPS-induced secretion of nitric oxide (NO), tumor necrosis factor-α (TNF-α), and Interleukin-1β (IL-1β). Moreover, Eriocitrin combined with resveratrol potently inhibited nuclear factor-κB (NF-κB), phosphor-STAT3, and phosphor-AKT, which was accompanied by inhibition of phosphorylation in mitogen-activated protein kinase (MAPK) signaling pathways. Treatment with eriocitrin combined with resveratrol alleviated edema and subcutaneous tissue inflammation caused by 12-O-tetradecanoylphorbol-13-acetate (TPA) in vivo. This study also demonstrated that treatment with eriocitrin and resveratrol decreased the levels of the pro-inflammatory cytokines TNF-α and IL-1β. The results of the present study indicate that eriocitrin combined with resveratrol effectively inhibits inflammatory responses both in vitro and in vivo.

The chemical compositions of essential oils (EOs) extracted from Curcuma kwangsiensis rhizomes collected from six natural habitats in P. R. China were evaluated using gas chromatography/mass spectrometry (GC/MS). Fifty‐seven components were identified from the six EOs, and their main constituents were 8,9‐dehydro‐9‐formyl‐cycloisolongifolene (2.37 – 42.59%), germacrone (6.53 – 22.20%), and l‐camphor (0.19 – 6.12%). The six EOs exhibited different DPPH radical‐scavenging activities (IC50, 2.24 – 31.03 μg/ml), with the activity of most of EOs being much higher than that of Trolox C (IC50, 10.49 μg/ml) and BHT (IC50, 54.13 μg/ml). Most EOs had potent antimicrobial effects against the tested bacteria and fungus. They also exhibited cytotoxicity against B16 (IC50, 4.44 – 147.4 μg/ml) and LNCaP cells (IC50, 73.94 – 429.25 μg/ml). The EOs showed excellent anti‐inflammatory action by significantly downregulating expression of pro‐inflammatory cytokines, cyclooxygenase‐2, and tumor necrosis factor‐α. This study provides insight into the interrelation among growth location, phytoconstituents, and bioactivities, and the results indicate the potential of C. kwangsiensis as natural nutrients, medicines, and others additives.

Chemical compositions, antioxidative, antimicrobial, anti-inflammatory, and cytotoxic activities of essential oils extracted from four common Curcuma species (Curcuma longa, Curcuma phaeocaulis, Curcuma wenyujin, and Curcuma kwangsiensis) rhizomes in P. R. China are comparatively studied. In total, 47, 49, 35, and 30 compounds are identified in C. longa, C. phaeocaulis, C. wenyujin, and C. kwangsiensis essential oils by GC/MS, and their richest compounds are ar-turmerone (21.67%), elemenone (19.41%), curdione (40.23%) and (36.47%), respectively. Moreover, C. kwangsiensis essential oils display the strongest DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging activity (IC50 , 3.47 μg/ml), much higher than ascorbic acid (6.50 μg/ml). C. phaeocaulis oils show the best antibacterial activities against Escherichia coli (MIC, 235.54 μg/ml), Pseudomonas aeruginosa (391.31 μg/ml) and Staphylococcus aureus (378.36 μg/ml), while C. wenyujin and C. kwangsiensis oils show optimum activities against Candida albicans (208.61 μg/ml) and Saccharomyces cerevisiae (193.27 μg/ml), respectively. C. phaeocaulis (IC50 , 4.63 μg/ml) and C. longa essential oils (73.05 μg/ml) have the best cytotoxicity against LNCaP and HepG2, respectively. C. kwangsiensis oils also exhibit the strongest anti-inflammatory activities by remarkably down-regulating expression of COX-2 and TNF-α. Therefore, due to their different chemical compositions and bioactivities, traditional Chinese Curcuma herbs should be differentially served as natural additives for food, pharmaceutical, and cosmetic.