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Objective:To investigate the changes in serum miRNA-21 and miRNA-4534 levels in patients with lung adenocarcinoma and their relationships with pathological characteristics and prognosis.Methods:A total of 148 patients with lung adenocarcinoma who received treatment in the First People's Hospital of Huzhou from January 2018 to January 2019 were included in the observation group. An additional 100 healthy controls who concurrently received physical examinations were included in the control group. The relative expression of serum miRNA-21 and miRNA-4534 was measured using real-time fluorescence quantitative PCR. The relative expression of serum miRNA-21 and miRNA-4534 was compared between the two groups. The relative expression of serum miRNA-21 and miRNA-4534 was compared between patients with different pathological characteristics. Total survival time was compared between patients with high and low miRAN-21 expression and between patients with high and low miRNA-4534 expression.Results:The relative expression of miRNA-21 and miRNA-4534 in the observation group was 3.27 ± 0.87 and 6.74 ± 1.56, respectively, which was significantly higher than 1.00 ± 0.01 and 1.02 ± 0.02 in the control group ( t = 20.07, 36.64, both P < 0.05). There was no significant difference in the relative expression of miRNA-21 and miRNA-4534 between different genders, ages, body mass index values, tumor regions, and tumor diameters (all P > 0.05). The relative expression of serum miRNA-21 in patients with stage Ⅲ-Ⅳ lung adenocarcinoma was 4.45 ± 1.05, which was significantly higher than 1.92 ± 0.53 in patients with stage Ⅰ-Ⅱ lung adenocarcinoma. The relative expression of serum miRNA-21 in patients with lymph node metastasis was higher than that in patients without lymph node metastasis (4.97 ± 1.28 vs. 1.34 ± 0.60, t = 17.53, 23.48, both P < 0.05). The relative expression of miRNA-4534 in patients with stage Ⅲ-Ⅳ lung adenocarcinoma was higher than that in patients with stage Ⅰ-Ⅱ lung adenocarcinoma (8.97 ± 1.85 vs. 4.48 ± 1.09, t = 17.17, P < 0.05). The relative expression of miRNA-4534 in patients with lymph node metastasis was higher than that in patients without lymph node metastasis (9.65 ± 1.35 vs. 3.78 ± 0.91, t = 30.59, P < 0.05). The total survival time in patients with high miRNA-21 expression was shorter than that in patients with low miRNA-21 expression [(18.37 ± 4.35) months vs. (29.93 ± 3.24) months, t = 18.45, P < 0.05]. The total survival time in patients with high miRNA-4534 expression was shorter than that in patients with low miRNA-4534 expression [(17.56 ± 4.21) months vs. (30.43 ± 3.63) months, t = 19.97, P < 0.05)]. Conclusion:Patients with lung adenocarcinoma have high expression of miRNA-21 and miRNA-4534 in the serum, which is closely related to the tumor-node-metastasis stage, lymph node metastasis, and prognosis.
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Endogenous ribonucleotides(RNs)and deoxyribonucleotides(dRNs)are important metabolites related to the pathogenesis of many diseases.In light of their physiological and pathological significances,a novel and sensitive pre-column derivatization method with N-(t-butyldimethylsilyl)-N-methyltri-fluoroacetamide(MTBSTFA)was developed to determine RNs and dRNs in human cells using high-performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS).A one-step extraction of cells with 85％methanol followed by a simple derivatization reaction within 5 min at room temper-ature contributed to shortened analysis time.The derivatives of 22 nucleoside mono-,di-and tri-phosphates were retained on the typical Cig column and eluted by ammonium acetate and acetonitrile in 9 min.Under these optimal conditions,good linearity was achieved in the tested calibration ranges.The lower limit of quantitation(LLOQ)was determined to be 0.1-0.4 μM for the tested RNs and 0.001-0.1 μM for dRNs.In addition,the precision(CV)was＜15％and the RSD of stability was lower than 10.4％.Furthermore,this method was applied to quantify the endogenous nucleotides in human colorectal carcinoma cell lines HCT116 exposed to 10-hydroxycamptothecin.In conclusion,our method has proven to be simple,rapid,sensitive,and reliable.It may be used for specific expanded studies on intracellular pharmacology in vitro.
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OBJECTIVE: To monitor and investigate the influencing factors of 6-methylmercaptopurine ribonucleotides (6-MMPR) in human red blood cell (RBC) in Chinese kidney transplant recipients treated with azathiopurine (AZA), and to provide references for clinical personalized medicine. METHODS: In 100 Chinese kidney transplant recipients, the concentration of 6-MMPR in RBC was detected by validated HPLC-UV method. Correlation analysis was performed to analyze the effects of various factors, including patient age, gender, weight, AZA dose and thiopurine S-methyltransferase (TPMT) activity, on the 6-MMPR concentration, and multivariate linear regression analysis was performed using SPSS20.0 software to investigate the influencing factors of 6-MMPR concentration. RESULTS: The concentration of 6-MMPR in 100 Chinese kidney transplant recipients was not normally distributed (P0.05), however, there was a significant positive correlation between TPMT activity and 6-MMPR concentration in RBC (P<0.001). CONCLUSION: TPMT activity in RBC is an independent factor affecting 6-MMPR concentration, which in turn affects the clinical efficacy and toxicity of the drug.
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The enhancer effect of glutamate monosodium (MSG) flavor was evaluated and its synergistic action with 5'-ribonucleotides: ionone rib nucleotides 5'-monophosphate (IMP) and guano sine monophosphate (GMP) in dehydrated soups consisting of lentils and peas. Four formulations were developed for both soups: the first was the target with the original level of MSG, the following had different concentrations and mixtures of these enhancers (6 percent MSG; 6 percent MSG and 0.26 percent IMP; 0.6 MSG and 0.12 percent IMP-GMP). A five-.point Graphic Hedonic Scale test was used, where 1 represented the most upset face and 5 represented the happiest face. The most accepted soup was selected by thirty elderly adults. The lentils soup with 0,6 MSG and 0J2 percent IMP-GMP and the pea's soup with 6 percent MSG and 0.26 percent IMP obtained the greatest level of acceptance. So, the effectiveness of the synergistic action between the MSG and 5'-ribonucleotides was demonstrated, because they can improve the acceptance of the evaluated formulation.
Se evaluó el efecto realzador del sabor del glutamato monosódico (GMS) y su acción sinergista con 5'-ribonucleótidos: inosinato monofosfato (IMP) y guanilato monofosfato (GMP), cuando se adicionaron a sopas deshidratadas de lentejas y arvejas. Se elaboraron 4 formulaciones para cada sopa, la primera formulación correspondió al control con su nivel de GMS original, las siguientes formulaciones contaron con distintas concentraciones y mezclas de estos realzadores (6 por ciento GMS; 6 por ciento GMS mas 0,26 por ciento IMP y 0,6 GMS mas 0,12 por ciento IMP-GMP). Se utilizó la evaluación sensorial de Escala Hedónica Gráfica, con una escala de 1 al 5, donde 1: representa "la carita más disgustada" y 5: "la más feliz". Treinta adultos mayores determinaron la formulación más aceptada. La sopa de lentejas con 6 por ciento de GMS mas 0,12 por ciento de IMP-GMP fue la que tuvo mayor aceptación, mientras que para la sopa de arvejas fue aquella que contenía 6 por ciento de GMS más 0,26 por ciento de IMP. Por tanto, se pudo demostrar la efectividad de la acción sinergista entre el GMS y los 5'-ribonucleótidos, al mejorar las aceptación de las formulaciones evaluadas.