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CHROMATOGRAPHIA · 2024 ·Vol. 87 ·期 6 ·407-418 · IF 1.2

Integration of Magnetic Dispersive Solid-Phase Micro-extraction with HPLC for Precise Detection of Tumor Markers 1-Methyladenosine and 1-Methylinosine in Urine Samples from Breast Cancer Patients

Hong Lei,Mao Huihui,Huang Xiaowei,Fu Xinyang,Lin Yanfang

To analyze 1-methyladenosine (m1A) and 1-methylinosine (m1I) in urine samples obtained from patients with breast cancer and healthy people, we used magnetic dispersive solid-phase micro-extraction (MDSPME) combined with high-performance liquid chromatography. The structure and morphology of magnetic nanoparticles, Fe 3 O 4 @GO, were investigated by FT-IR, XRD, TEM, and TGA. m1A and m1I in urine samples were enriched by the solid phase micro-extraction technology of Fe 3 O 4 @GO. Several parameters affecting the extraction efficiency, including adsorption time, pH, ionic strength, mass of adsorbent, extraction time, eluent solvent, elution time, and elution temperature, were researched and optimized. Under the optimized conditions, the corresponding linearity range of the two analytes showed a good linearity ( r 2  > 0.9998) and significant addition recoveries ranging from 88.50 to 109.38%, while the limit of detection was between 0.10 and 0.11 ng/mL. The enrichment factors of m1A and m1I were 108 and 134, respectively. The method was simple, sensitive, and reliable for the detection and purification with good detection limit, and it was suitable for the detection of m1A and m1I in urine samples. In addition, the analysis of modified nucleosides is of great value for the early diagnosis of patients with breast cancer.

研究领域: 分析化学 · 肿瘤学 · 医学诊断
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES · 2023 · IF 8.2

Chitosan and its oligosaccharide accelerate colonic motility and reverse serum metabolites in rats after excessive protein consumption

Linlin Wang,Lilong Wang,Cui Cao,Jun Zhao,Chen Song,Zhijie Bao,Chunhong Yan,Shuang Song

Excessive protein consumption (EPC) could increase the gastrointestinal burden and impair gut motility. The present study was designed to explore the improvement of chitosan (CTS) and chitosan oligosaccharide (COS) on colonic motility and serum metabolites in rats after EPC. The results of in vivo experiments fully proved that CTS and COS could improve gut motility and reverse the serum metabolites in rats as indicated by LC-MS/MS analysis, and the COS group even showed a better effect than the CTS group. Furthermore, short-chain fatty acids (SCFAs), which could promote gut motility, were also increased to alleviate EPC-induced constipation after supplementation with CTS or COS. In addition, CTS and COS could decrease the concentration of ammonia in serum and down-regulate the levels of H 2 S and indole. In summary, the present study revealed that CTS and COS could produce SCFAs, improve the colonic motility in rats, reverse the levels of valine , adenosine, cysteine, 1-methyladenosine, indole, and uracil , and enhance aminoacyl-tRNA biosynthesis and valine, leucine and isoleucine degradation. The present study provides novel insights into the potential roles of CTS and COS in alleviating the adverse effects of EPC.

研究领域: 营养学 · 胃肠病学 · 代谢组学
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Redox Biology · 2017 · IF 6.337

Gene and protein expressions and metabolomics exhibit activated redox signaling and wnt/β-catenin pathway are associated with metabolite dysfunction in patients with chronic kidney disease

Dan-Qian Chen,Gang Cao,Hua Chen,Dan Liu,Wei Su,Xiao-Yong Yu,Nosratola D. Vaziri,Xiu-Hua Liu,Xu Bai,Li Zhang,Ying-Yong Zhao

Changes in plasma concentration of small organic metabolites could be due to their altered production or urinary excretion and changes in their urine concentration may be due to the changes in their filtered load, tubular reabsorption, and/or altered urine volume. Therefore, these factors should be considered in interpretation of the changes observed in plasma or urine of the target metabolite(s). Fasting plasma and urine samples from 180 CKD patients and 120 age-matched healthy controls were determined by UPLC-HDMS-metabolomics and quantitative real-time RT-PCR techniques. Compared with healthy controls, patients with CKD showed activation of NF-κB and up-regulation of pro-inflammatory and pro-oxidant mRNA and protein expression as well as down-regulation of Nrf2-associated anti-oxidant gene mRNA and protein expression, accompanied by activated canonical Wnt/β-catenin signaling. 124 plasma and 128 urine metabolites were identified and 40 metabolites were significantly altered in both plasma and urine. Plasma concentration and urine excretion of 25 metabolites were distinctly different between CKD and controls. They were related to amino acid, methylamine, purine and lipid metabolisms. Logistic regression identified four plasma and five urine metabolites. Parts of them were good correlated with eGFR or serum creatinine. 5-Methoxytryptophan and homocystine and citrulline were good correlated with both eGFR and creatinine. Clinical factors were incorporated to establish predictive models. The enhanced metabolite model showed 5-methoxytryptophan, homocystine and citrulline have satisfactory accuracy, sensitivity and specificity for predictive CKD. The dysregulation of CKD was related to amino acid, methylamine, purine and lipid metabolisms. 5-methoxytryptophan, homocystine and citrulline could be considered as additional GFR-associated biomarker candidates and for indicating advanced renal injury. CKD caused dysregulation of the plasma and urine metabolome, activation of inflammatory/oxidative pathway and Wnt/β-catenin signaling and suppression of antioxidant pathway.

研究领域: 肾病学 · 代谢组学 · 分子生物学 · 临床化学
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ANALYTICAL CHEMISTRY · 2018 · IF 6.042

Enrichment and Quantitative Determination of 5-(Hydroxymethyl)-2′-deoxycytidine, 5-(Formyl)-2′-deoxycytidine, and 5-(Carboxyl)-2′-deoxycytidine in Human Urine of Breast Cancer Patients by Magnetic Hyper-Cross-Linked Microporous Polymers Based on Polyionic Liquid

Mengzhe Guo,Liyan Zhang,Yan Du,Wencheng Du,Dantong Liu,Cheng Guo,Yuanjiang Pan,Daoquan Tang

5-(Hydroxymethyl)-2′-deoxycytidine (5-hmdC), 5-(formyl)-2′-deoxycytidine (5-fodC), and 5-(carboxyl)-2′-deoxycytidine (5-cadC) are crucial intermediate products of the DNA demethylation pathway, which can also act as potential biomarkers reflecting the diagnosis and prognosis in multiple tumors. Detecting 5-hmdC, 5-fodC, and 5-cadC in human urine has various advantages including readily available samples and being noninvasive to patients. However, few works have reported the detection of 5-fodC and 5-cadC due to their trace amounts. Here we developed a novel magnetic hyper-cross-linked microporous polymer (HMP) material based on polyionic liquid (PIL) for the enrichment of 5-hmdC, 5-fodC, and 5-cadC. These magnetic PIL-HMP materials provided specific enrichment superiority for three modified cytidines. After enrichment, the signal intensity of 5-hmdC, 5-fodC, and 5-cadC increased 10–75-fold with lower limits of quantitation (LLOQ) of 0.049, 0.781, and 0.781 ng/mL, respectively. The recoveries were approximately 86.5–95.2% for 5-hmdC, 95.2–107.8% for 5-fodC, and 99.4–102.4% for 5-cadC under the relative standard deviation (RSD) of 0.2–10.3%. Finally, we successfully applied magnetic PIL-HMP materials coupled with high-performance liquid chromatography–electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) in enrichment and quantitative determination of 5-hmdC, 5-fodC, and 5-cadC in human urine of 10 breast cancer patients and 10 healthy people. We found that the level of 5-hmdC decreased in breast cancer patients (p < 0.05), while the levels of 5-fodC and 5-cadC increased (p < 0.05, p < 0.01). Our results demonstrated that the levels of metabolic 5-hmdC, 5-fodC, and 5-cadC in human urine are closely related to breast cancer, which could contribute to the clinical diagnosis and investigation of breast cancer and its occurrence and development mechanisms.

研究领域: 医学 · 分析化学 · 肿瘤学
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