Ultra-performance Liquid Chromatography-mass Spectrometry Research Articles

Gut bacteria Prevotellaceae related lithocholic acid metabolism promotes colonic inflammation

BackgroundThe conversion of primary bile acids to secondary bile acids by the gut microbiota has been implicated in colonic inflammation. This study investigated the role of gut microbiota related bile acid metabolism in colonic inflammation in both patients with inflammatory bowel disease (IBD) and a murine model of dextran sulfate sodium (DSS)-induced colitis.MethodsBile acids in fecal samples from patients with IBD and DSS-induced colitis mice, with and without antibiotic treatment, were analyzed using ultraperformance liquid chromatography-mass spectrometry (UPLC-MS). The composition of the microbiota in fecal samples from IBD patients and DSS-colitis mice was characterized via Illumina MiSeq sequencing of the bacterial 16S rRNA gene V3-V4 region. Metagenomic profiling further identified metabolism-related gene signatures in stool samples from DSS-colitis mice. Histological analysis, quantitative PCR (qPCR) and Western Blotting were conducted on colonic samples from DSS-induced colitis mice to assess colonic inflammation, mucosal barrier integrity, and associated signaling pathways. The multivariate analysis of bile acids was conducted using Soft Independent Modelling of Class Analogy (SIMCA, Umetrics, Sweden). The relation between the relative abundance of specific phyla/genera and bile acid concentration was assess through Spearman's correlation analyses. Finally, lithocholic acid (LCA), the key bile acid, was administered via gavage to evaluate its effect on colonic inflammation and mucosal barrier integrity.ResultsIn patients with IBD, the composition of colonic bile acids and gut microbiota was altered. Moreover, changes in the gut microbiota further modulate the composition of bile acids in the intestine. As the gut microbiota continues to shift, the bile acid profile undergoes additional alterations. The aforementioned alterations were also observed in mice with DSS-induced colitis. The study revealed a correlation between dysbiosis of the gut microbiota and modifications in the profile of colonic bile acids, notably LCA observed in both patients with IBD and mice with DSS-induced colitis. Through multivariate analysis, LCA was identified as the key bile acid that significantly affects colonic inflammation and the integrity of mucosal barrier. Subsequent experiments confirmed that LCA supplementation effectively mitigated the inhibitory effects of gut microbiota on colitis progression in mice, primarily through the activation of the sphingosine-1-phosphate receptor 2 (S1PR2)/NF-κB p65 signaling pathway. Analysis of the microbiome and metagenomic data revealed changes in the gut microbiota, notably an increased abundance of an unclassified genus within the family Prevotellaceae in DSS-induced colitis mice. Furthermore, a positive correlation was observed between the relative abundance of Prevotellaceae and bile acid biosynthesis pathways, as well as colonic LCA level.ConclusionsThese findings suggest that LCA and its positively correlated gut bacteria, Prevotellaceae, are closely associated with intestinal inflammation. Targeting colonic inflammation may involve inhibiting LCA and members of the Prevotellaceae family as potential therapeutic strategies.

Centipeda minima extracts and the active sesquiterpene lactones have therapeutic efficacy in non-small cell lung cancer by suppressing Skp2/p27 signaling pathway.

Centipeda minima (L.) A. Braun & Asch (C. minima) was applied to treat nasal allergy, headache, cough, and even nasopharyngeal carcinoma in traditional Chinese medicine. However, the underlying anticancer mechanisms of C. minima and its active components have not been systematically illustrated. The study aims to examine the therapeutic efficacy of the ethanol extract of C. minima (ECM) and its active components in non-small cell lung cancer (NSCLC) and illustrate the underlying mechanisms. The main chemical components in the ethanol extract of C. minima (ECM) and the supercritical CO2 fluid extract of C. minima (CM-SFE) were determined by using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). The antitumor effects of ECM and CM-SFE were examined by using NSCLC cell xenografts. The flow cytometry, cell colony formation, wound-healing, transwell assay, and Western blotting were conducted to investigate the anticancer properties of ECM, CM-SFE, and these sesquiterpene lactones that abundantly distributed in these extracts. We first determined that ECM contains high levels of sesquiterpene lactones. ECM can markedly induce cell cycle arrest and suppress migration and invasion of NSCLC cells. Mechanistically, ECM promoted proteasome-dependent degradation of Skp2 protein and induced the accumulation of its substrates p27; whereas Skp2 overexpression can attenuate the inhibitory effects of ECM on NSCLC proliferation and migration. Moreover, ECM at 200-600mg/kg can significantly inhibit tumor growth and metastasis in A549-luciferase cell orthotopic xenografts by suppressing Skp2 expression. The sesquiterpene lactones that abundantly distributed in ECM, including 6-O-angeloylplenolin (6-OAP), arnicolide D (ArD) and arnicolide C (ArC), were also demonstrated to decrease Skp2 while increase p27 protein level, thereby significantly inducing cell cycle arrest and suppressing migration of NSCLC cells. Notably, CM-SFE, which mainly consisted of 6-OAP, ArD and ArC, exhibited much stronger anti-NSCLC activity than that of ECM in A549-luciferase cell orthotopic xenografts. Our results demonstrate that the active components in C. minima possesses potential anti-NSCLC activities by suppressing Skp2/p27 signaling pathway, and these active sesquiterpene lactones can be further developed as potent Skp2 inhibitor to treat NSCLC.

Protective effects of Euphorbia heterophylla against testicular degeneration in streptozotocin-induced diabetic rats in relation to phytochemical profile.

Diabetes mellitus (DM) poses a major risk to human health due to an array of implications, one of which is a detrimental effect on the testicular and reproductive functions. Euphorbia heterophylla is widely recognized for its medicinal properties worldwide. The objective of this study was to profile E. heterophylla ethanol extract (EH-EtOH) and elucidate its protective role in oxidative stress, relieving inflammatory action of hyperglycemia-induced testicular degeneration and restoring the normal histological structure with physiological properties of testicular tissue in streptozotocin (STZ)-induced DM. High-resolution ultra-performance liquid chromatography-mass spectrometry (UHPLC-ESI-Orbitrap-MS) analysis was employed to analyze the compounds present in EH-EtOH. The protective effect of EH-EtOH against testicular degeneration in the rat model of DM was evaluated by measuring improvements in blood glucose levels, body weight, testicular inflammation, oxidative damage, testicular microcirculation impairment, and apoptosis of testicular cells induced by STZ. The chemical profiling of EH-EtOH revealed the presence of 52 compounds, including phenolic acids, flavonoids, coumarins, phloroglucinols, and triterpenes. Notably, this study identified isovitexin-C-hexoside, isorhamnetin-O-hexoside, diosmetin, and halfordin for the first time in Euphorbia species. Treatment with EH-EtOH effectively mitigated the damage caused by STZ, as evidenced by restored testosterone (T4) levels and antioxidant capacity, reduced expression of pro-inflammatory cytokines, improved testicular microcirculation, and inhibition of apoptosis in the testes. These results emphasize the potential therapeutic effect of E. heterophylla on DM related to male infertility and reproductive dysfunctions via its antioxidant/angiogenetic /anti-apoptotic effect.

Untargeted metabolite profiling reveals metabolic disorders in livers of rats with mepiquat exposure.

Mepiquat is a contaminant produced in thermal-processed food. It can induce spleen and liver injury. However, the mechanism that mepiquat induced hepatotoxicity remains unclear. In this study, a ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS)-based metabolomic analysis of livers in rats was performed to explore metabolic alterations. Our results demonstrated that there were 36 differential metabolites. Eleven major disordered metabolic pathways were found. Network analysis of differential metabolites and metabolic pathways indicated that glutamic acid was a possible key upregulated metabolite. It participated in nine metabolic networks. Glutathione and l-proline may be key downregulated metabolites. They were involved in eight and seven metabolic pathways, respectively. Compared with the control group, serum concentrations of alanine aminotransferase, aspartate aminotransferase, and total bile acid in high dosage group increased significantly. In addition, histopathological analysis showed liver injury in rats with mepiquat exposure. These data were consistent with results of metabolomics. Our results offered new insights for molecular mechanism of liver toxicity induced by mepiquat. PRACTICAL APPLICATION: Mepiquat is a contaminant formed in thermal-processed food. It can induce spleen and liver injury. Our results offered new insights for molecular mechanism of liver toxicity induced by mepiquat. It will provide important information for official limits of mepiquat in foods.

Potential Mechanisms and Effects of Dai Bai Jie Ethanol Extract in Preventing Acute Alcoholic Liver Injury

This study investigated the protective effect of Dai Bai Jie (DBJ) extract against acute alcoholic liver injury (AALI) and elucidated its potential mechanism. The total saponin level in the DBJ extracts was measured using vanillin–chloroform acid colorimetry. To observe the preventive and protective effects of DBJ on AML-12 cells in an ethanol environment, the effective components of DBJ were identified. An alcohol-induced AALI mouse model was used to evaluate the efficacy of DBJ against AALI. For this purpose, alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) levels were assessed, liver function indices and oxidative and inflammatory markers were determined, and histopathological examinations were performed. Mechanistic investigations were conducted using RT-qPCR assays and immunohistochemical analysis to determine the protective effects of DBJ. The samples (DBJ-1, DBJ-2, and DBJ-3) were obtained by extracting DBJ with water, 50% ethanol, and 95% ethanol, yielding total saponin contents of 5.35%, 6.64%, and 11.83%, respectively. DBJ-3 was isolated and purified, and its components were identified by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS). DBJ-3 had the greatest effect on cell viability in an ethanol environment. Moreover, DBJ-3 reduced inflammatory infiltration, liver cell degeneration, and hemorrhage, while increasing ADH and ALDH levels in liver tissues. Additionally, DBJ-3 considerably decreased the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), and triglyceride (TG) levels. DBJ-3 reduced malondialdehyde (MDA), reactive oxygen species (ROS), and inflammatory factors, such as tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), and interleukin 6 (IL-6), while increasing superoxide dismutase (SOD) and glutathione S-transferase (GST) activities. Furthermore, DBJ-3 significantly increased alcohol dehydrogenase 1b (ADH1B) and aldehyde dehydrogenase 2 (ALDH2) expression at the gene and protein levels within alcohol metabolism pathways and reduced the nuclear factor kappa-B (NF-κB) gene and protein levels. These findings suggest that DBJ-3 can prevent AALI by enhancing alcohol metabolism via the regulation of ADH1B and ALDH2 and the modulation of the NF-κB pathway to improve antioxidant and anti-inflammatory effects.

Pharmacokinetic Interaction between Imatinib and Tacrolimus in Rats

Objective: Tacrolimus, a calcineurin inhibitor (CNI), is the first-line treatment for chronic myeloid leukemia (CML) and advanced gastrointestinal stromal tumors (GIST). Imatinib and tacrolimus are both substrates of the hepatic enzymes CYP3A4/5 and efflux transporter P-gp, so drug-drug interactions may occur during their co-administration treatment. Therefore, this study aimed to evaluate the pharmacokinetic interaction between imatinib and tacrolimus in rats. Methods: Rats were divided into groups I (30 mg/kg imatinib administered for 14 days), II (1.89 mg/kg tacrolimus and 30 mg/kg imatinib administered for 14 days), III (30mg/kg imatinib and 0.63mg/kg tacrolimus administered for 14 days), IV (1.89mg/kg tacrolimus for 14 days), and V (10mg/kg imatinib and 1.89mg/kg tacrolimus for 14 days). Blood samples were determined for whole blood of tacrolimus, plasma of imatinib, and Ndesmethyl imatinib concentrations using ultra-performance liquid chromatography-mass spectrometry. Results: After 1 day of a single dose, tacrolimus had no significant effect on the pharmacokinetics of imatinib and N-desmethyl imatinib; imatinib significantly increased the AUC and Cmax of tacrolimus (P < 0.05). After 14 days of multiple doses, tacrolimus significantly reduced the AUC and Cmax of imatinib and N-desmethyl imatinib (P < 0.05). Further, imatinib significantly increased AUC0-24 and AUC0-∞ of tacrolimus (P < 0.05). Conclusion: Imatinib increased tacrolimus blood concentrations after single and multiple administrations. Tacrolimus did not significantly affect the pharmacokinetics of imatinib after a single dose; however, tacrolimus might impact the absorption and metabolism of imatinib after multiple doses. The results showed that when imatinib and tacrolimus were co-administered, attention should be paid to the presence of drug-drug interactions.

Kalanchoe crenata Andrews (Haw.) Improves Losartan's Antihypertensive Activity.

Cardiovascular diseases constitute one of the leading causes of morbidity and mortality worldwide. Herbal medicines represent viable alternatives to the synthetic drugs currently employed in the control of hypertension. This study aimed to isolate and identify the chemical markers of Kalanchoe crenata and to investigate the antihypertensive and anti-matrix metalloproteinase (MMP2) activities of an aqueous extract of the leaves. The main constituents of the aqueous extract of K. crenata were separated by ultra-performance liquid chromatography-mass spectrometry, and their presence was identified by NMR spectroscopy. Renovascular hypertension was induced in male Wistar rats using the two-kidney one-clip method (HTN groups), while control animals (Sham groups) were submitted to Sham surgery. Six groups of 10 animals each were treated daily for eight weeks as follows: Sham 1 (carrier), Sham 2 (K. crenata extract), HTN.1 (carrier), HTN.2 (K. crenata extract), HTN 3 (losartan), and HTN 4 (K. crenata extract with losartan). The main compounds of the extract were patuletin 3-O-(4″-O-acetyl-α-L-rhamnopyranosyl)-7-O-(3‴-O-acetyl-α-L-rhamnopyranoside) (1), patuletin 3-O-α-L-rhamnopyranosyl-7-O-L-rhamnopyranoside (2), and trans-caffeoyl-malic acid (3), with compounds 1 and 2 being chemical markers of the species. Significant reductions (p < 0.05) in systolic blood pressure and MMP2 (72kDa isoform) activity were observed in the HTN 4 group. The association of K. crenata extract and losartan presented in vivo effects against hypertension.

Alterations in the metabolic signature of full-term infants of diabetic mothers

BackgroundMaternal diabetes during pregnancy may alter the metabolomic profile of the offspring, increasing the risk of perinatal complications and potentially predisposing them to adverse metabolic consequences during later life. To assess this risk, we aimed to study the levels of amino acids and acylcarnitines in the cord blood of infants of diabetic mothers (IDMs).MethodsFifty term-IDMs were compared to 25 healthy newborns. Of the diabetic mothers, 37 had gestational diabetes mellitus (GDM), and 13 had pre-gestational diabetes mellitus (PGDM). Amino acid and acylcarnitine concentrations were measured in dried spot samples of cord blood using ultra-performance liquid chromatography–mass spectrometry (UPLC-MS).ResultsWe observed significantly higher levels of valine, aspartic acid, the leucine-to-isoleucine (Leu:Ile), and leucine-to-phenylalanine (Leu:Phe), and decreased values of methionine and the phenylalanine-to-tyrosine (Phe:Tyr) in neonates in the GDM group compared to controls. Neonates of mothers with GDM showed higher levels of C2-carnitine, C5-carnitine, C5:1, and C4-OH (C3-DC) compared to controls. Neonates of mothers with PGDM also showed higher levels of C2-carnitine, C0-carnitine, and C5:1 compared to controls. There were no significant differences in amino acids or acylcarnitine levels between IDMs with GDMs and PGDMs. Principle component analysis (PCA) showed that principal component 1 (PC1) was responsible for the majority of differences between IDMs and controls. PC1 consisted of aspartic, valine, leu:lle, C4-OH(C3-DC), C2-carnitine, C0-carinitine, C5-carnitine, C18:1, and C16:1. Variable importance in projection (VIP) scores based on PLS-DA loadings showed high values for aspartic, valine, leu:lle, Leu:Phe, C4-OH(C3-DC), C2-carnitine, C0-carnitine, C5:1, C5-DC, C18:1, C16:1, C5-carnitine in IDMs, and high values for Met:Phe, methionine, and Phe:Tyr in controls.ConclusionMaternal diabetes leads to alterations in amino acid concentrations and the carnitine shuttle pathway of their offspring.

Chemometric Classification and Bioactivity Correlation of Black Instant Coffee and Coffee Bean Extract by Chlorogenic Acid Profiling.

Chlorogenic acids (CGAs) play a key role in defining the quality and functionality of coffee products. CGA fingerprints of black instant coffee (BIC) and coffee bean extract (CBE) were profiled using ultra-performance liquid chromatography-mass spectrometry and analyzed by chemometrics. A total of 25 CGAs were identified. The BICs yielded higher levels of major CGAs than the CBEs. Furthermore, chemometrics methods, including principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA), successfully classified the CBEs and the BICs. In vitro cellular antioxidant activity and viability assays between coffee products further confirmed the relationship between phenolic compounds and bioactivities. Compared to the CBEs, the BICs provided higher cellular toxicity and oxidant activity for hepatocellular carcinoma G2 (HepG2) cells. These results demonstrated that CGAs and their derivatives could be markers for studying coffee-related products. This study revealed the unique phenolic profiles of various coffee products, highlighting the differences between whole beans and soluble coffee.

Spatial Distribution, Source Analysis, and Risk Assessment of Multiple Antibiotics in Chaohu Lake Basin

Chaohu Lake is the fifth largest freshwater lake in China that meets the needs of surrounding industries for water, tourism, and flood storage, making it important for the socio-economic development and modernization of Anhui Province. As an important ecological wetland in the lower reaches of the Yangtze River, many rare bird habitats are present along the coast of Chaohu Lake. Therefore, it is eco-economically important to study antibiotic pollution in Chaohu Lake. To unveil antibiotic contamination in Chaohu Lake, surface water and sediment samples were collected from the main basin of Chaohu Lake by steamship in June 2022. Solid phase extraction and ultra-performance liquid chromatography-mass spectrometry （SPE-UPLC-MS） was used to determine the occurrence of 31 target antibiotics, including tetracyclines, sulfonamides, macrolides, and quinolones. The study focused on the spatial distribution, source analysis, and risk assessment of various antibiotics in the surface water and sediments of the main basin of Chaohu Lake. The occurrences of 20 different antibiotics were successively identified in the surface water and sediments of Chaohu Lake. These antibiotics included sulfonamides, quinolones, macrolides, tetracyclines, and others. Significant spatial differences in antibiotics were found between surface water and sediments. The horizontal spatial distribution results indicated differences in species and concentration of antibiotics in surface water and sediments. The vertical spatial distribution results showed that many types of antibiotics had consistent vertical distribution characteristics in surface water and sediments at the same point. The principal component analysis results indicated that the source of antibiotics in surface water was similar to that in sediment and considered with the actual distribution of existing industries around Chaohu Lake； the main sources were livestock and poultry breeding sewage, aquaculture sewage, and household sewage. The ecological risk quotient （RQ） results indicated that norfloxacin in the surface water of Chaohu Lake posed a high risk to algae （RQ &gt; 1）, while sulfamethoxazole and doxycycline posed a medium risk to algae （0.1 ≤ RQ &lt; 1）. The health risk quotient （HRQ） showed that the risk of exposure to antibiotics through drinking water was very low （HRQ &lt; 0.01）. Our findings provide scientific support for decision makers to control antibiotic pollution and reduce the management costs of antibiotic watershed pollution.

Pharmacological effects and mechanisms of danlong oral liquid in asthma airway remodeling: Insights from serum medicinal chemistry, network pharmacology, and experimental validation.

Danlong oral liquid (DLOL) is a traditional Chinese proprietary medicine commonly used to treat chronic respiratory diseases, including bronchial asthma and chronic obstructive pulmonary disease. However, the therapeutic effects and pharmacological mechanisms of DLOL in improving airway remodeling remain unclear. This study utilizes in vivo and in vitro experiments, serum pharmacological analysis, and network-based pharmacology approaches to investigate the effects and mechanisms of DLOL on airway remodeling and epithelial-mesenchymal transition (EMT) in asthma. An asthma model was established through ovalbumins (OVA) sensitization and challenge in BALB/c mice to observe the effects of DLOL on airway hyperresponsiveness (AHR), inflammation, remodeling, and molecular markers of EMT. The absorbed chemical prototype constituents of DLOL were analyzed using Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS), and targets for asthma and airway remodeling were predicted using a network pharmacology approach. Key biological processes and signaling pathways were analyzed. Additionally, TGF-β1 was used to induce EMT in BEAS-2B cells. TGF-β1 and DLOL-containing serum were screened to determine the optimal time and concentration in BEAS-2B cells using CCK8 assays. The cell scratch assay was used to assess cell migration, while immunofluorescence and immunohistochemistry were employed to evaluate protein expression levels. DLOL improved AHR in asthmatic mice, reduced inflammatory cell infiltration in lung tissue, decreased airway wall and smooth muscle thickness, and reduced collagen deposition. It also down-regulated mesenchymal markers (N-cadherin, vimentin, α-SMA) and key remodeling factors (TGF-β1, MMP9), while up-regulating the epithelial marker E-cadherin. A total of 17 absorbed chemical prototype constituents were identified, predicting 54 core targets involved in airway remodeling. Following Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, the key targets were found to be associated with the regulation of cell migration, cell-cell adhesion, and cell adhesion molecular processes, with the PI3K-Akt signaling pathway likely playing a critical role. Cellular experiments confirmed that DLOL-containing serum inhibited TGF-β1-induced EMT in BEAS-2B cells and suppressed the phosphorylation of Akt and GSK-3β. This study identifies, for the first time, the serum medicinal chemistry of DLOL using UPLC-MS. Combining network pharmacology, in vivo and in vitro experiments, it elucidates the effects and potential mechanisms of the drug on airway remodeling and EMT. DLOL may offer a novel therapeutic approach for asthma-related airway remodeling.

Apixaban removal during emergency surgery for type A acute aortic dissection: a prospective cohort study.

Acute type A aortic dissection (ATAAD) has a high mortality, and acute aortic repair is the only curative treatment. In patients treated with factor Xa (FXa) inhibitors, the risk of severe disease-related complications such as cardiac tamponade and hemodynamic shock must be balanced against the potential for severe perioperative bleeding. The aim was to study intraoperative changes in plasma levels of the FXa inhibitor apixaban when using hemoadsorption during acute thoracic aortic repair. This is a single-center prospective cohort study. Eight apixaban-treated patients presenting with ATAAD underwent acute thoracic aortic repair with intraoperative hemoadsorption with CytoSorb. Apixaban concentrations were measured at the start of cardiopulmonary bypass (CPB) and after 5, 15, 30, 60, and 90min of CPB, at CPB weaning, 30min after CPB weaning and 24h postoperatively, using ultraperformance liquid chromatography-mass spectrometry (UPLC-MS). After 30min of CPB with hemoadsorption, mean apixaban concentration (±SD) was reduced by 59% from 108 (±69)µg/l to 44 (±20)µg/l (P=0.009). There was a further reduction to 37 (±17)µg/l at CPB weaning (P=0.008). Apixaban concentration displayed an increase to 56 (±29)µg/l 24h postoperatively (P=0.01). In-hospital mortality was 25%. The mean 24H chest tube drainage volume was 621 (±136)ml. Intraoperative hemoadsorption lowers apixaban levels in patients undergoing emergency surgery for ATAAD. Further research is needed to determine its impact on perioperative bleeding complications and mortality.

Pre-diagnostic plasma advanced glycation end-products and soluble receptor for advanced glycation end-products and mortality in colorectal cancer patients.

Advanced glycation end-products (AGEs), formed endogenously or obtained exogenously from diet, may contribute to chronic inflammation, intracellular signaling alterations, and pathogenesis of several chronic diseases including colorectal cancer (CRC). However, the role of AGEs in CRC survival is less known. The associations of pre-diagnostic circulating AGEs and their soluble receptor (sRAGE) with CRC-specific and overall mortality were estimated using multivariable-adjusted Cox proportional hazards regression among 1369 CRC cases in the European Prospective Investigation into Cancer and Nutrition (EPIC) study. Concentrations of major plasma AGEs, Nε-[carboxy-methyl]lysine (CML), Nε-[carboxy-ethyl]lysine (CEL) and Nδ-[5-hydro-5-methyl-4-imidazolon-2-yl]-ornithine (MG-H1), were measured using ultra-performance liquid chromatography mass-spectrometry. sRAGE was assessed by enzyme-linked immunosorbent assay. Over a mean follow-up period of 96 months, 693 deaths occurred of which 541 were due to CRC. Individual and combined AGEs were not statistically significantly associated with CRC-specific or overall mortality. However, there was a possible interaction by sex for CEL (Pinteraction = .05). Participants with higher sRAGE had a higher risk of dying from CRC (HRQ5vs.Q1 = 1.67, 95% CI: 1.21-2.30, Ptrend = .02) or any cause (HRQ5vs.Q1 = 1.38, 95% CI: 1.05-1.83, Ptrend = .09). These associations tended to be stronger among cases with diabetes (Pinteraction = .03) and pre-diabetes (Pinteraction <.01) before CRC diagnosis. Pre-diagnostic AGEs were not associated with CRC-specific and overall mortality in individuals with CRC. However, a positive association was observed for sRAGE. Our findings may stimulate further research on the role of AGEs and sRAGE in survival among cancer patients with special emphasis on potential effect modifications by sex and diabetes.

Magnetic recyclable g-C3N4/CuFe2O4/MnO2 activated peroxymonosulfate process via dual Z-scheme heterojunction for photodegradation of ciprofloxacin

Composite heterojunctions effectively enhance the separation efficiency of photoelectrons and holes, thereby activating the peroxymonosulfate (PMS) process to degrade recalcitrant pollutants. Consequently, a g-C3N4/CuFe2O4/MnO2 (CN/CFO/MN) composite with a dual Z-scheme heterojunction was developed for the first time and coupled with PMS for the degradation of ciprofloxacin. CN/CFO/MN exhibited strong binding ability to ciprofloxacin with an adsorption capacity of 38mg/g, reaching adsorption equilibrium within 60min with high efficiency, which is crucial for effective degradation. Additionally, isolated CN/CFO/MN and bare PMS (0.3g/L) demonstrated limited degradation capabilities with ciprofloxacin removal rate of 51% and 41% under illumination, respectively. Notably, the elimination efficiency of ciprofloxacin was enhanced to nearly 100% by integrating CN/CFO/MN photodegradation with PMS activation. Furthermore, compared to pure CN, CFO, MN, and the binary CN/CFO, CN/CFO/MN provided superior photocatalytic degradation, improving the removal efficiency from 45% to 63%. The dual Z-scheme heterojunction in CN/CFO/MN furnishes ample photoelectrons to activate the PMS process for the generation of free radicals. Active species exploration experiments suggest that the CN/CFO/MN-PMS-light method primarily degrades ciprofloxacin by catalyzing the generation of holes, superoxide, hydroxyl and sulfate radicals. The CN/CFO/MN-PMS-light method achieved complete removal of 0.02–0.1g/L ciprofloxacin within 5min in tap, pond and river water systems. Ultra-performance liquid chromatography-mass spectrometry analysis-determined the degradation pathways of ciprofloxacin to involve the piperazine ring, defluorination reaction, and oxidation reaction of quinolone. Assessment via toxicity estimation software indicated that the toxicity of the degradation products was significantly reduced reaching harmless level. This work demonstrates that the CN/CFO/MN-PMS-light method with a dual Z-scheme heterojunction holds considerable potential for environmental purification.

Mechanism of Colquhounia Root Tablets in inhibiting osteoclast differentiation based on HSP90 target modulation

This study aimed to investigate the potential role of Colquhounia Root Tablets against bone destruction in rheumatoid arthritis(RA) and its molecular mechanism. The study used ultra-performance liquid chromatography-mass spectrometry to analyze the major components of Colquhounia Root Tablets and predicted its candidate target gene set based on the major components. The key targets of RA bone destruction were obtained through GeneCards and the Database of Genetics and Medical Literature(OMIM), protein-protein interaction(PPI) network was constructed, and the key targets were identified by topological analysis. The molecular mechanism of Colquhounia Root Tablets against RA bone destruction was further revealed using Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis. The effects of Colquhounia Root Tablets on macrophage viability was assessed by MTS assay and screened for non-toxic concentrations. A model of receptor activator of nuclear factor-κB(RANKL) induced osteoclast differentiation in vitro was constructed. Colquhounia Root Tablets were used to observe the formation and differentiation of osteoclasts by tartrate-resistant acid phosphatase(TRAP) staining and fibrous actin(F-actin) staining, and the effects of Colquhounia Root Tablets on the changes of core target proteins in the osteoclast differentiation system were detected by immunofluorescence and Western blot. The results showed that the main components of Colquhounia Root Tablets included 14 compounds such as triptolide, celastrol, and triptophenolide. Further network analysis revealed that heat-shock protein 90(HSP90) was the key target gene of Colquhounia Root Tablets for anti-RA bone destruction. TRAP staining and F-actin staining showed that the number and area of TRAP-positive polymorphonuclear cells, as well as actin rings, were reduced in a dose-dependent manner after the intervention of Colquhounia Root Tablets(P&lt;0.01). Western blot results showed that the expression of HSP90 protein was significantly reduced after intervention with Colquhounia Root Tablets at 20 and 40 μg·mL~(-1)(P&lt;0.01); Colquhounia Root Tablets at 10 μg·mL~(-1) could significantly decrease the expression of necrosis factor receptor associated molecule 6(TRAF6) and nuclear factor of activated T cells 1(NFATc1) proteins(P&lt;0.01); moreover, all doses of Colquhounia Root Tablets significantly reduced the expression of osteoclast differentiation marker proteins matrix metalloproteinase 9(MMP9) and cathepsin K(CTSK)(P&lt;0.01).Immunofluorescence results further confirmed that Colquhounia Root Tablets significantly inhibited HSP90 and CTSK levels, as well as NFATc1 activation in osteoblasts. In conclusion, the present study confirmed that Colquhounia Root Tablets may inhibit RANKL-induced osteoclast differentiation by regulating the key target of HSP90, thus exerting an anti-RA bone destruction effect, which will provide a new idea for Colquhounia Root Tablets to prevent and treat bone destruction in rheumatoid arthritis.

Volatile organic compounds (VOC) metabolites in urine are associated with increased systemic inflammation levels, and smokers are identified as a vulnerable population

BackgroundPrevious studies indicated that exposure to VOCs was linked to increased systemic inflammation levels. However, the dose-response relationships between urine VOCs metabolites and systemic inflammation have not been established, and the key metabolite of the toxic compounds has not been identified. MethodsWe used data in 7007 US adults in the NHANES cycles (2011–2018) across 8 years. Urinary VOC metabolites were measured using ultra-performance liquid chromatography and electrospray tandem mass spectrometry (UPLC-ESI/MSMS). VOC metabolites were adjusted by urinary creatinine level before analysis. Systemic inflammation was assessed by systemic immune-inflammation index (SII) and systemic inflammation response index (SIRI) indices. Generalized linear models, restricted cubic splines (RCS), and weighted quantile sum (WQS) regression were applied to evaluate the associations, exposure-response (E-R) curve and identify the key contributor compound, adjusting for gender, age, race, BMI, marital condition, education level, smoking level, alcohol consumption and physical activity. Smoking status was assessed as an effect modifier. ResultsSignificant and robust positive correlations were found between 8 VOC metabolites and both SII and SIRI. They were N-Acetyl-S-(2-carboxyethyl)-L-cysteine (CEMA), N-Acetyl-S-(2-cyanoethyl)-L-cysteine (CYMA), N-Acetyl-S-(3,4-dihydroxybutyl)-L-cysteine (DHBMA), N-Acetyl-S-(3-hydroxypropyl)-L-cysteine (3HPMA), mandelic acid (MA), N-Acetyl-S-(4-hydroxy-2-butenyl)-L-cysteine (MHBMA3), phenylglyoxylic acid (PGA), and N-Acetyl-S-(3-hydroxypropyl-1-methyl)-L-cysteine (HPMMA). The RCS curves showed J-shaped or exponential shaped E-R relationships for most VOC metabolites. WQS regression found that exposure to the mixture of VOC metabolites was related to increased systemic inflammation, and MA was the key VOC metabolite contributing most to systemic inflammation levels. Smokers exhibited higher levels of urinary VOCs and larger susceptibility to VOC-related increases in SII and SIRI compared to non-smokers. ConclusionThis study demonstrated a strong link between urinary VOC metabolites and increased systemic inflammation, and smokers were more susceptible. Our findings highlighted the significance of reducing VOC exposure to mitigate the inflammation levels, particularly for smokers.

Inactivation of CYP2D6 by Berberrubine and the Chemical Mechanism.

Berberrubine (BRB), belonging to the benzylisoquinoline alkaloid, is a main metabolite of berberine in vivo. BRB was previously proven to undergo metabolic activation mediated by P450s. In this study, the chemical interactions between BRB and CYP2D6 enzyme were investigated. First, a variety of P450s participated in the metabolism of berberine transformed to BRB, but CYP2D6 was the most involved enzyme. A time-, concentration-, and nicotinamide adenine dinucleotide phosphate (NADPH)-dependent inhibition of CYP2D6 was caused by BRB. The inhibitory effect of BRB on CYP2D6 was irreversible. The maximum reaction rate constants of inactivation (kinact) and half-maximal inactivation (KI) of BRB on CYP2D6 were 0.0410 min-1 and 3.798 μM, respectively. Metoprolol, a classic substrate of CYP2D6, attenuated CYP2D6 from inactivation by BRB. Glutathione (GSH) and catalase/superoxide dismutase failed to protect against the inactivation of CYP2D6 caused by BRB. Three cys-based adducts derived from the reaction of electrophilic metabolites of BRB with CYP2D6 were detected by ultra performance liquid chromatography-mass spectrometry (UPLC-MS)/MS. The reactive metabolites derived from BRB might be responsible for the inactivation of CYP2D6. In summary, BRB was characterized as a mechanism-based inactivator of CYP2D6.

Effect of Coptidis Rhizoma on gastrointestinal system before and after processing with wine based on gut microbiota and short chain fatty acids.

Coptis deltoidea C.Y. Cheng et Hsiao (CD), commonly used in the treatment of heat-toxin congestion and excessiveness. However, CD needs to be processed with wine for alleviating the bitter and cold of CD, meanwhile, reducing the gastrointestinal damage. The research assessed the discrepant effects of CD on gastrointestinal system before and after processing with wine, and explore the potential mechanisms. The ingredients in CD and CD processed with wine (PCD) were performed on Ultra Performance Liquid Chromatography Mass Spectrometry (UPLC-MS). The mice were treated with CD and PCD once a day for 6weeks (0.65 and 2.6g/kg, i.g.). The pathological changes of gastrointestinal tract were evaluated, and the serum inflammatory factors and Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) RelA (p65) protein of tissues were determined. The short chain fatty acids (SCFAs) of feces were analyzed by UPLC-MS, the gut microbiota (GM) changes were performed on 16S rRNA sequencing. Ingredients analysis declared that the alkaloids, flavonoids, phenylpropanoid compounds were the main metabolites in CD and PCD. CD reduced body weight and food intake, and the effect of CD on fecal water content increased first and then decreased with the prolongation of administration time, while its effect on intestinal transport time was exactly the opposite, reduced the SCFAs contents of feces. CD caused different degree of damage to the gastrointestinal tract, and the effect on the small intestine and colon was more obvious, which increased the expression of NF-κB p65 and elevated the inflammatory factors levels. PCD were weaker than that of CD. In addition, CD and PCD can change the composition of GM, and reduced the levels of Lactobacillus, Allobaculum, Ruminococcus, and norank_f_S24-7, increased the levels of Akkermansia, Dorea, Bacteroides, and Blautia at the genus level. However, PCD induced a milder effect of GM dysregulation than that of CD. CD can cause damage to the gastrointestinal tract, which may be related to the GM disorders, SCFAs changes-mediated by GM, abnormal NF-κB p65 expression and increased inflammatory factors levels, interestingly, PCD had a lower effect than CD, which may be related to the differences in the types and contents of ingredients in CD after processing. And this study provided data support for the mechanism of processing with wine to alleviate "bitter-cold injury the stomach" of CD.

Exposure to Volatile Organic Compounds and Blood Pressure in NHANES 2011 to 2018.

Volatile organic compounds (VOCs) are ubiquitous environmental pollutants. Exposure to VOCs is associated with cardiovascular disease risk factors, including elevated blood pressure in susceptible populations. However, research in the general population, particularly among nonsmoking adults, is limited. We hypothesized that higher VOC exposure is associated with higher blood pressure and hypertension, among nonsmokers. We included 4 cycles of data (2011-2018) of nonsmoking adults (n=4430) from the National Health and Nutrition Examination Survey. Urinary VOC metabolites were measured by ultraperformance liquid chromatography-mass spectrometry, adjusted for urine dilution, and log-transformed. We estimated mean differences in blood pressure using linear models and prevalence ratio of stage 2 hypertension using modified Poisson models with robust standard errors. Models were adjusted for age, sex, race and ethnicity, education, body mass index, estimated glomerular filtration rate, and National Health and Nutrition Examination Survey cycle. Participants were 54% female, with a median age of 48 years, 32.3% had hypertension, and 7.9% had diabetes. The mean differences (95% CI) in systolic blood pressure were 1.61 (0.07-3.15) and 2.46 (1.01-3.92) mm Hg when comparing the highest with the lowest quartile of urinary acrolein (N-acetyl-S-[2-carboxyethyl]-L-cysteine) and 1,3-butadiene (N-acetyl-S-[3,4-dihydroxybutyl]-L-cysteine) metabolites. The prevalence ratios for hypertension were 1.06 (95% CI, 1.02-1.09) and 1.05 (95% CI, 1.01-1.09) when comparing the highest with lowest quartiles of urinary acrolein (N-acetyl-S-[2-carboxyethyl]-L-cysteine) and 1,3-butadiene (N-acetyl-S-[3,4-dihydroxybutyl]-L-cysteine), respectively. Exposure to VOCs may be a relevant yet understudied environmental contributor to cardiovascular disease risk in the nonsmoking, US population.

Impact of Drying on Phytonutritional Compounds, In Vitro Antioxidant Activity and Cytotoxicity of Spiny Saltbush (Rhagodia spinescens).

The Spiny saltbush (Rhagodia spinscens) is a halophyte species with the potential to provide natural ingredients used in food and pharmaceutical industries. In food and pharmaceutical applications, drying is necessary to maintain shelf-life, which reduces phytonutrient content. In this study, changes in the nutritional composition, phenolic and carotenoid profiles of radical antioxidant scavenging activity [(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)], antioxidant power [ferric reducing antioxidant ability assay (FRAP)], and cytotoxicity of freeze- and oven-dried (55 °C for 24 h) spiny saltbush were determined. Sodium (4.72 g/100 g dry weight (DW), potassium (6.86 g/100 g DW), calcium (4.06 g/100 g DW), zinc (372 mg/kg DW) and protein content were higher in oven-dried samples than freeze-dried samples. Ultra-performance liquid chromatography-mass spectrometry analysis detected 18 metabolites in saltbush extracts. Partial Least-Squares Discriminant Analysis, Hierarchical Cluster Analysis, and Variable Importance in Projection discriminated between freeze-dried and oven-dried samples. Freeze-dried samples retained more individual metabolites than oven-dried samples, while oven-dried samples had higher antioxidant activity (ABTS and FRAP), lutein, trans-β carotene, and cis-β-carotene. Correlation analysis identified potential antioxidant candidates between phenolic and carotenoid compounds. Neither freeze-dried nor oven-dried spiny saltbush samples showed cytotoxicity. The study uncovered changes in phytonutritional compounds after the oven and freeze-drying spiny saltbush, a potential salt alternative and functional ingredient for the food industry.