Optically levitated micro- and nanoparticles are an ideal optomechanical platform for precision measurements, particularly enabling the detection of ultraweak forces. Nevertheless, quantum backaction and inherent instabilities induced by the trapping laser fundamentally restrict further improvements in force sensitivity and resolution. To circumvent these bottlenecks, we actively drive the levitated nanoparticle's mechanical motion in a phase-locked phonon laser mode and integrate a carrier-modulation measurement architecture to enhance force sensing capabilities. The stable and high-amplitude oscillation of the phonon laser allows for the robust trapping under 1 mW-level laser power, which in turn reduces the force noise to 4.0(3)*10^-22 N/Hz^1/2. Furthermore, by using phase-locked phonon laser, the measurement system achieves active stabilization and extended coherence time with the measured signal to 12,500 seconds, realizing a measurement resolution of 8(4)*10^-24 N with a sensitivity of 9.3(7)*10^-22 N/Hz^1/2 under a loaded force. These results establish the phonon laser as a low-noise, long-coherence-time, self-stabilizing platform for precision measurements, as well as in quantum and fundamental physics tests.

Chronic kidney disease (CKD) is a major global health burden, with renal fibrosis as the key pathological driver of disease progression. Fucoidan (FPS) is a sulfated polysaccharide from brown algae that has demonstrated renoprotective and anti-fibrotic properties. However, its metabolic and microbiota-related mechanisms remain unclear. Here, we evaluated the efficacy of FPS in a unilateral ureteral obstruction (UUO) mouse model, in which mice received oral FPS (100 or 200 mg kg-1 day-1) for 14 days. Kidney metabolomics and 16S rRNA sequencing of the gut microbiota were performed. As a result, UUO induced pronounced disturbances in tryptophan metabolism and marked gut microbial dysbiosis. FPS treatment attenuated renal fibrosis, normalized key tryptophan pathway intermediates and related metabolic enzymes in the kidneys, and partially restored the gut microbial composition. These results implicate the modulation of tryptophan metabolism and the gut microbiota in the anti-fibrotic effects of FPS, supporting its potential as a natural therapeutic candidate for renal fibrosis and CKD.

The use of irinotecan (CPT-11), a first-line chemotherapeutic agent for colorectal cancer (CRC), is limited by acquired drug resistance and severe adverse effects. It has been found that activation of farnesoid X receptor (FXR) signaling reduced the growth of CRC. Here, we investigated the combination of obeticholic acid (OCA), a well-characterized FXR agonist, with CPT-11 as a potential therapeutic combination for CRC. In vitro experiments demonstrated that the OCA-CPT-11 synergistically inhibited proliferation and migration in FXR-high HT-29 and SW620 cells, but exhibited minimal synergy in FXR-low HCT116 and Caco-2 cells. Genetic knockdown of FXR in HT-29 cells attenuated the synergistic effects, whereas FXR overexpression in HCT116 cells enhanced them. These findings indicate that the synergistic effect of OCA-CPT-11 depends on basal FXR expression levels, highlighting the need to identify both FXR inducers and agonists. Through literature and database screening, silibinin (SB) and nigakinone (Nig) were identified to upregulate and activate FXR. Both SB-CPT-11 and Nig-CPT-11 exhibited strong synergistic anti-CRC effects in vitro, even under low FXR conditions. Nig achieved synergy with CPT‑11 at lower effective concentrations than SB, so it was prioritized as the lead candidate for in vivo evaluation. Consistently, the Nig-CPT‑11 combination synergistically inhibited the growth of HCT116 subcutaneous xenograft tumors in vivo. Notably. In summary, combining FXR inducers and agonists with CPT-11 offers a promising strategy for the treatment of CRC.

Lactylation is a widespread protein modification with important regulatory roles in health and disease. Emerging evidence also links metabolite lactylation to disease onset, progression, and treatment response. It is estimated that an uncharacterized pool of lactoyl-metabolites exists in vivo, according to its formation mechanism. To mine this pool, we combined knowledge-driven prediction with stable isotopic labeling-based high-resolution mass spectrometry (SIL-HRMS). A predictive library of 151 lactoyl-metabolites was generated by expanding 20 basic amino acids (AAs) to their upstream and downstream metabolites. To improve detection, we developed a pair of new hydroxyl-reactive labeling reagents, 3,5-(dimethylamino)-2,4,6-triazine benzene-1-chlorine (Tmt-aycl-Cl) and its deuterium form d12-Tmt-aycl-Cl. Using HRMS, 95 lactoyl-metabolites were putatively identified across 14 tissue types, 64 structures of which are reported for the first time. Representative lactoyl-metabolites were chemically synthesized and characterized to verify some of the annotations. We then developed a pseudotargeted metabolomics workflow for semiquantitative profiling in a diabetic kidney disease (DKD) mouse model. DKD mice showed elevated lactate in serum and the kidney and widespread dysregulations of lactoyl-metabolites. Many lactoyl-metabolites correlated strongly with the urinary albumin-to-creatinine ratio, suggesting biomarker potential for DKD. This integrated strategy expands the catalog of lactoyl-metabolites and provides a platform for investigating their biological significance.

ETHNOPHARMACOLOGICAL RELEVANCE:Dachaihu Decoction (DCHD) is a traditional Chinese medicine formula from the Shanghan Lun. It has been used for over 1800 years to treat "Shaoyang and Yangming concurrent syndromes", which are closely related to modern liver disorders, including metabolic dysfunction-associated steatohepatitis (MASH). While DCHD has demonstrated hepatoprotective effects in preclinical studies, the specific bioactive components and mechanism of action against MASH remain elusive. AIM OF THE STUDY:To systematically identify the key active components of DCHD mediating its anti-MASH effects and to elucidate their molecular mechanisms. MATERIALS AND METHODS:The components in DCHD were characterized using high resolution-mass spectrometry (HRMS). The potential bioactive components were screened using network pharmacology. The therapeutic efficacy of candidate compounds was evaluated both in free fatty acid (FFA)-induced hepatocyte injury and activated hepatic stellate cells (HSCs) and in a high-fat diet/CCl4-induced MASH mouse model. The molecular mechanism was investigated via transcriptomic profiling, drug-target interaction validation, and gene knockdown assays. RESULTS:A total of 128 components were identified from DCHD using HRMS. Lonicerin (LON), a flavonoid glycoside predominantly derived from Citrus trifoliata L., was identified as the key active component. LON significantly ameliorated steatosis, inflammation and fibrosis both in vitro and in vivo. Mechanistically, LON bound to lymphocyte antigen 6 family member D (LY6D), which was significantly upregulated during MASH progression, and dose-dependently suppressed its expression. Importantly, LY6D knockdown abolished the protective effect of LON in AML12 hepatocytes and activated HSCs, confirming LY6D as a functional target. CONCLUSIONS:This study identified LON as a key bioactive component mediating the anti-MASH effects of DCHD and revealed that it alleviated MASH progression by targeting LY6D.

Multidrug resistance (MDR) is a significant challenge in cancer treatment, with limited effective strategies available. Neuropilin-1 (NRP1) is emerging as a potential therapeutic target for overcoming drug resistance, but its role in MDR and the identification of potential inhibitors require further exploration. In this study, we investigated the role of NRP1 in MDR and identifies potential inhibitors targeting NRP1. Elevated NRP1 expression was observed in oxaliplatin (OXP)-resistant HCT116 (HCT116/L) and cisplatin (DDP)-resistant A549 cells (A549/DDP). Virtual screening and biological assays identified pitavastatin (Ptv) as a potent NRP1 inhibitor that restored chemosensitivity in resistant cells both in vitro and in vivo. Mechanistic studies revealed that Ptv directly binds to NRP1, promotes degradation of Zinc finger X-chromosomal protein (ZFX), and disrupts the NRP1-ZFX axis to reverse MDR. This study provides promising prospects for targeting the NRP1-ZFX axis as a therapeutic strategy for MDR and highlights the potential clinical application of Ptv in diseases involving NRP1.

Cisplatin (DDP) is widely utilized in the clinical treatment of malignant tumors, but its effectiveness is significantly compromised by the adverse effects of acute kidney injury (AKI). Renal tubular cells are primarily responsible for DDP-induced AKI (DDP-AKI); however, the responses of heterogeneous renal tubular cells to DDP exposure have not been thoroughly explored. In this study, we employed a targeted metabolomics approach to investigate the metabolic responses of renal tubular cells in DDP-AKI rats. Tubular cells were isolated from the renal cortex and outer medulla, and a chemical derivatization-based liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics method was applied. Our findings revealed distinct metabolic profiles in tubular cells from the renal cortex and outer medulla, with outer medullary cells exhibiting greater sensitivity to DDP exposure. Further analyses identified the tryptophan pathway as a critical factor contributing to these regional differences. Additional functional investigations showed that intermediate metabolites of the tryptophan pathway alleviated DDP cytotoxicity in both cortical and outer medullary tubular cells primarily through modulation of the Bcl2/Bax and Caspase-3 pathway. This study enhances our understanding of the metabolic characteristics of tubular cells across heterogeneous renal regions in DDP-AKI and facilitates further exploration of the underlying mechanisms of DDP-induced nephrotoxicity.

Ulcerative colitis (UC) is an idiopathic, chronic inflammatory disorder with an increasing incidence worldwide. Due to the complex and unclear therapeutic targets, unmet UC therapeutic drugs still exist. Recently, acylcarnitine metabolism disorder has been linked to intestinal inflammation, but its role in UC remains elusive. According to our preliminary non-targeted metabolomics data, acylcarnitines (ACs) was screened as the disturbed metabolites in the different intestinal inflammation-related diseases. Here we quantified 26 ACs within liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the dextran sulfate sodium (DSS)-induced UC rat model, and found that long-chain acylcarnitines (LCACs) were increased to varying degrees. As the key metabolites of fatty acid β-oxidation (FAO), the upstream metabolites long-chain fatty acids (LCFAs) and the related metabolic enzymes were further characterized, the results showed that the rate-limiting enzyme carnitine palmitoyltransferase 1A (CPT1A)-mediated LCFAs-LCACs metabolic axis was activated sharply. Next in vitro experiments exhibited that CPT1A was significantly upregulated in both inflammatory macrophages and colonic epithelial cells, and inhibition or knockdown of CPT1A could reduce the inflammation level remarkably. Thus, we screened the pharmacologic inhibitors of CPT1A from US Food and Drug Administration (FDA) approved drugs, within molecular docking, Western blot and cell membrane chromatography (CMC) technology, gliquidone was found to inhibit CPT1A in a dose-dependent manner and exert anti-inflammatory effects in vitro. Animal experiments also showed that gliquidone alleviated DSS-induced UC significantly. In summary, our study presents that within metabolomics analysis, inhibiting CPT1A is focused to be a potential therapeutic strategy against UC, and gliquidone represents an alternative treatment.

Environmental pollutants can induce multiorgan damage, with the digestive tract particularly susceptible. Diabetic enteropathy is a significant complication of type 2 diabetes mellitus (T2D). However, the relationship between environmental pollutant exposure and T2D-associated intestinal injury has not been previously explored. In this study, T2D mice were subjected to polystyrene microplastics (PS-MPs, 100 μg/day, 3 weeks) and bisphenol A (BPA, 100 μg/kg/day, 2 weeks). Metabolomics and 16S rRNA sequencing were used to detect changes in colonic metabolites and gut microbial composition. Caco-2 cells were utilized to investigate the functions of the altered metabolites. Compared to the T2D group, mice exposed to PS-MPs and BPA exhibited shorter colon length and reduced levels of gut barrier proteins ZO-1 and Occludin. Metabolomics analysis revealed that PS-MPs primarily affected colonic long-chain fatty acids (LCFAs) and adenosine metabolism, while BPA disrupted α-ketoisovaleric acid (KIVA) and pyruvic acid (PyrA) homeostasis. Moreover, PS-MPs exposure altered the abundance of Duncaniella and Olsenella, while BPA primarily affected Phocaeicola, Olsenella, and Variovorax. In vitro experiments showed that palmitoleic acid (C16:1), γ-linolenic acid (C18:3), adenosine (Ado), and KIVA promoted the expression of ZO-1 in Caco-2 cells. Our findings provide valuable insights into the impact of environmental pollutants on intestinal injury in T2D, underscoring the importance of environmental contaminant management, particularly in susceptible populations.

BACKGROUND:Ulcerative colitis (UC) has demonstrated an escalating global incidence and prevalence, thereby posing substantial challenges to public health. Despite recent advancements in therapeutic interventions, the clinical management of UC remains suboptimal, underscoring the urgent need for novel treatment strategies. Saikosaponin A (SSa), a bioactive compound derived from the traditional Chinese herb Radix Bupleuri (RB), exhibits potent anti-inflammatory and immunomodulatory effects. However, its effects on UC and the underlying molecular mechanisms remain to be thoroughly explored. PURPOSE:This study aims to elucidate the underlying mechanisms of SSa in ameliorating UC and establish a pharmacological foundation for developing novel treatment modalities to address unmet clinical needs in UC treatment. METHODS:The protective effects of SSa against DSS-induced acute colitis were evaluated in a 3 % DSS-treated mouse model. Histological (H&E staining) and molecular analyses (RT-qPCR, ELISA, Western blotting, and flow cytometry) were performed to assess colonic tissue damage and inflammatory responses. Macrophage depletion via tail vein injection of clodronate liposomes confirmed the pivotal role of macrophages in UC pathogenesis and SSa's anti-inflammatory effects. The inhibitory effects of SSa on NLRP3 inflammasome activation were analyzed in vivo and in LPS/ATP-stimulated bone marrow-derived macrophages (BMDMs) using RT-qPCR, ELISA, and Western blotting. Bioinformatics analysis, targeted LC-MS/MS, and molecular docking were employed to identify potential molecular targets and mechanisms of SSa. Drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and CH25H siRNA knockdown assays were used to validate CH25H as the direct target of SSa. RESULTS:SSa effectively attenuated DSS-induced colitis in mice by alleviating colonic inflammation, preserving intestinal barrier integrity, reducing LPS translocation, and mitigating systemic organ injury in the liver and spleen. The inflammatory response of macrophages and the production of IL-1β were identified as key pathogenic components in colitis, and the clearance of macrophages significantly ameliorated colitis progression while SSa administration post-macrophage clearance did not further alter the disease phenotype. Mechanistically, SSa inhibited the NLRP3 inflammasome activation-mediated IL-1β secretion in macrophages. The sterol metabolism played a crucial role in the activation of the NLRP3 inflammasome. SSa also restored the disturbed sterol homeostasis in the colon under inflammatory conditions, especially promoted the synthesis of 25-hydroxycholesterol (25-OHC). Further investigation revealed that SSa primarily exerts its therapeutic effects by directly targeting cholesterol 25-hydroxylase (CH25H), which promotes the production of 25-OHC and inhibits macrophage NLRP3 inflammasome activation. CONCLUSION:This pioneering study demonstrated the therapeutic effect of SSa on DSS-induced colitis by targeting CH25H to enhance 25-OHC biosynthesis, which subsequently inhibited NLRP3 inflammasome activation in IL-1β-producing macrophages. These findings reveal a novel mechanism of SSa in UC treatment through cholesterol metabolism-regulated cascade immune modulation, providing strong pharmacological support for its development as a potential UC therapy.

We theoretically investigate the problem of position detection of an optically levitated Mie particle. The information radiation field (IRF) is proposed and defined to characterize the scattered light carrying complete information about the center-of-mass (c.m.) motion of the particle. Based on the IRF, we suggest an optimal detection scheme for the position of arbitrary particles. We calculate both the information losses of objective collection and mode-matching in levitated optomechanical experiments. Our results conclude that the backward detection scheme, using an incident Gaussian beam focused by a high numerical aperture lens, provides sufficient information to achieve the quantum ground state through cooling of the three-dimensional c.m. motion of the Mie particle.

Lung adenocarcinoma (LUAD), the predominant subtype of non-small cell lung cancer (NSCLC), poses significant therapeutic challenges due to its aggressive nature and limited treatment options. Cisplatin is a commonly used chemotherapeutic agent for advanced LUAD, often encounters PDR, leading to rapid disease progression and tumor recurrence. In this study, we demonstrate that LDHA expression is elevated in cisplatin-resistant LUAD cell lines and correlates with poor patient prognosis. Notably, inhibiting the expression of LDHA significantly enhances cisplatin sensitivity. Mechanistically, LDHA functions through non-metabolic enzymatic activity to promote the ubiquitination and degradation of AMBRA1 by facilitating its interaction with RNF2, thereby activating Cyclin D1 and BCL2, which drives cisplatin resistance. Importantly, we identified two LDHA inhibitors, triflupromazine (TRI) and tranylcypromine (TRA), that significantly improve cisplatin efficacy both in vitro and in vivo. These findings highlight a critical role of LDHA in promoting cisplatin resistance, and suggest that targeting LDHA may represent a promising therapeutic strategy for patients with advanced LUAD.

Phosphatidyl amino acids (p-AAs) are metabolites characterized by the phosphorylation of the hydroxyl, amino, carboxyl, and thiol groups of amino acids. Previous research has primarily focused on the phosphorylation sites within macromolecular proteins, with a particular emphasis on typical O-p-AAs. In this study, we established a prediction library of p-AAs based on existing knowledge. To improve detection rates of p-AAs in biological samples, we employed a chemical labeling-based LC-MS/MS method, utilizing p-[3,5-(dimethylamino)-2,4,6-triazine] benzene-1-sulfonyl piperazine (Tmt-PP) and its deuterated form (d12-Tmt-PP) as paired labeling reagents. A preliminary identification was performed by matching characteristic MS fragments with available standards. Additionally, strategies such as in vitro methods were implemented for further identification. The phosphatase treatment aids in identifying phosphate-modified metabolites by dephosphorylating them, while cell extract incubation helps determine if novel phosphorylated amino acids are generated in vivo. Ultimately, we identified 11 p-AAs, 6 of which are novel metabolites reported for the first time. A pseudotargeted metabolomics method covering 11 identified p-AAs was established and applied to investigate the differences between cisplatin-resistant non-small cell lung cancer (NSCLC) cells and their parental cells, as well as their derived exosomes. This approach enhances our understanding of the role of p-AAs in various health and disease conditions and contributes to the discovery of additional novel phosphatidyl metabolites.

Lung adenocarcinoma (LUAD), the most widely existing subtype of non-small cell lung cancer (NSCLC), is a leading cause of cancer-related mortality, characterized by challenging early diagnosis, high rates of recurrence and metastasis, and poor prognosis. Chemotherapy remains the primary treatment for advanced LUAD, but its effectiveness is often hindered by the development of chemoresistance. In this study, a targeted metabolomics method unveiled a marked up-regulation of glycolysis in chemotherapy-resistant LUAD cells. Particularly, the ratio of fructose 1,6-bisphosphate (FBP) to fructose 6-phosphate (F6P) reflected the activity of the rate-limiting enzyme Phosphofructokinase muscle isoform (PFKM) was significantly elevated. We further observed a significant increase in exosome release in chemotherapy-resistant cells. More importantly, it was found that the interaction between PFKM and exosomes plays a role in regulating chemoresistance in LUAD. Mechanistically, PFKM influences exosomes release by modulating Ras-related protein Rab-8B (RAB8B) expression, impacting apoptosis and glycolytic metabolism, thereby promoting chemoresistance. Furthermore, drug-resistant cells enhance chemoresistance in sensitive cells by releasing exosomes with heightened glycolytic activity. These findings highlight the crucial role of the PFKM-RAB8B axis in promoting chemoresistance, suggesting it as a potential therapeutic target for countering LUAD chemoresistance.

Colorectal cancer (CRC) is the third most common malignancy globally and the second leading cause of cancer-related mortality. Its development is a multifactorial and multistage process influenced by a dynamic interplay between gut microbiota, environmental factors, and fatty acid metabolism. Dysbiosis of intestinal microbiota and abnormalities in microbiota-associated metabolites have been implicated in colorectal carcinogenesis, highlighting the pivotal role of microbial and metabolic interactions. Fatty acid metabolism serves as a critical nexus linking dietary patterns with gut microbial activity, significantly impacting intestinal health. In CRC patients, reduced levels of short-chain fatty acids (SCFAs) and SCFA-producing bacteria have been consistently observed. Supplementation with SCFA-producing probiotics has demonstrated tumor-suppressive effects, while therapeutic strategies aimed at modulating SCFA levels have shown potential in enhancing the efficacy of radiation therapy and immunotherapy in both preclinical and clinical settings. This review explores the intricate relationship between gut microbiota, fatty acid metabolism, and CRC, offering insights into the underlying mechanisms and their potential translational applications. Understanding this interplay could pave the way for novel diagnostic, therapeutic, and preventive strategies in the management of CRC.

Free fatty acids (FFAs) play a key role in living organisms and participate in metabolic processes, mainly as a source of energy for cells. The CC position isomer of FFAs is strongly associated with many diseases, but quantification of its double bond position isomer remains a challenge in lipid analysis. In this study, based on the requirements of the pseudotargeted metabolomics, an analytical method based on a two-step derivatization LC-MS/MS method and a multiple reaction monitoring (MRM) mode was established and fully validated in terms of linearity, precision, and stability. The results demonstrated that the method relatively quantified 30 FFAs in plasma samples. These included 8 saturated fatty acids (SFAs), 12 monounsaturated fatty acids (MUFAs), and 10 polyunsaturated fatty acids (PUFAs). The developed method was applied to analyze the increase or down-regulation of the relative content of FFAs before and after the administration of the drug in beagles which may help to explain the therapeutic effect of sacubitril valsartan. The establishment of this method allows the study of FFAs in beagles plasma to be no longer limited to analysis at the subclass level but can be focused at the isomer level.

Advanced glycation end products (AGEs), formed through reactions between carbonyl compounds and amino groups, mostly originate from lysine and arginine, with other amino-acid-derived AGEs remaining largely uncharacterized. Here, we developed a comprehensive strategy to annotate novel AGEs. First, a prediction library of 216 AGEs was constructed based on reported formation pathways. Available standards were labeled by dansyl chloride to obtain characteristic fragments via liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using Maillard reaction models of heated amino acid and carbonyl mixtures, combined with LC-MS/MS and high-resolution MS, we identified 20 AGEs, including 7 new structures. A pseudotargeted method covering these AGEs was developed and applied to study their dynamic changes in herbs subjected to varying steam-bask cycles and to investigate differences between free and protein-bound AGEs in high-temperature processed foods. This strategy advances the discovery and understanding of diverse AGEs in food and herbal matrices.

Metabolomics reflects the physiological states of organisms exposed to distinct environmental stimuli. However, little is known about the metabolic signatures of rainbow trout under different temperatures. In the present study, a 56-day experiment was conducted to evaluate the growth performance and metabolic signatures of liver tissues in rainbow trout (Oncorhynchus mykiss) juveniles exposed to seawater at 7 degrees C (T7), 11 degrees C (T11), 15 degrees C (T15) and 19 degrees C (T19), nearly covering the temperature ranges in aquaculture industry during different seasons. The results indicated that individuals cultured at 15 degrees C showed growth performance with significantly higher final weight, weight gain, and specific growth rate than the others. Furthermore, a total of 24 liver samples (6 replicated samples x 4 treatment groups) were performed for non-targeted metabolomics analysis, resulting in 4852 and 5688 effective ions detected by ESI+ and ESI- modes, respectively. Of them, 474 and 425 metabolites were systematically identified and annotated in ESI+ and ESI- modes, respectively. Nearly half of metabolites (43.94%, 395) were classified as lipids and lipid-like molecules in HMDB database. Based on the strict thresholds, a total of 150, 66 and 45 differential metabolites (DMs) were determined in three different comparisons, including T7 vs. T15, T11 vs. T15 and T19 vs. T15. It was noted that these DMs could be further grouped into 4 clusters according to their abundance tendencies as temperatures increased from 7 degrees C to 19 degrees C. The relative abundance of DMs associated with sphingolipid metabolism was continuously increased with the elevated temperatures, while a decreasing tendency was observed in these DMs related to glycerophospholipid meta-bolism. Meanwhile, DMs in phenylalanine and histidine metabolism displayed the highest abundance at 15 degrees C, and N-acyl taurine and vitamin metabolism was induced by high temperature (19 degrees C). Taken together, different temperatures strongly affected both amino acid and lipid metabolism in liver tissues of rainbow trout. The study provides insight into the potential physiological basis for the better cultivation and fisheries management of rainbow trout at different temperatures.