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Objective:To analyze the serum and urinary amino acid (AA) profiles of urolithiasis patients to explore the potential biomarkers for clinical screening and early diagnosis.Methods:Case-control study. Serum and urine samples were collected from 74 urolithiasis patients (aged 20-82 years, 41 men, 33 female) in the department of urology of the First Affiliated Hospital of Fujian Medical University and 35 healthy controls (HC, aged 22-80 years old, 20 men, 15 female) from the health examination center from February 2015 to October 2017. Serum and urinary AA levels of patients and HC were analyzed using Gas Chromatography-Mass Spectrometry (GC-MS) based metabolomic strategy. The multivariate statistical analysis methods of principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) were employed for modeling. The variable importance projection (VIP) value of OPLS-DA model>1 and P<0.05 of t test were selected to screen the differential amino acid metabolites. The diagnostic capabilities of potential markers were evaluated by receiver operating characteristic (ROC) curve and binary logistic regression analysis. Results:Five AA metabolites including serine, glutamate, aspartic acid, isoleucine and glycine were found, which had statistically significant differences between the patient group and the control group ( P<0.05) and were associated with seven metabolic pathways. Serum serine, glutamate, aspartic acid, isoleucine and urine glycine and aspartic acid were combined into an integrated marker panel whose AUC value was 0.890, the sensitivity was 78.0%, and the specificity was 96.4%. Conclusion:Five amino acids in serum and urine could be used as an integrated biomarker panel for the clinical screening and early diagnosis of urolithiasis, which could provide some experimental basis for molecular urolithiasis research.
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Objective To investigate the hepatoprotective effects of total flavonoids in Scorzonera austriaca Wild (TFSA) in vivo and in vitro. Methods In vivo, ICR mice were randomly divided into negative, model, positive, TFSA’s low-dose, medium-dose and high-dose groups,and acute chemical liver injury models were constructed with CCl4 and acute autoimmune liver injury models with Bacillus Calmette-Guerin vaccine ( BCG ) and lipopolysaccharide (LPS).The activity of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) was detected and liver tissue was used as biopsy.In vitro, liver cells of Wistar rat were extracted and isolated by orthotopic collagenase digestion method, and liver cell damage was induced with CCl4.Then the liver cells were cultured with TFSA solution and the contents of AST, ALT, LDH, nitric oxide (NO), superoxide dismutase (SOD) in the supernatant and malondialdehyde (MDA) in rat hepatocyte were detected.Results The results of CCl4-and BCG+LPS-induced acute chemical liver injury models in mice showed that there were less microstructures damage of liver tissue in TFSA groups compared with model group in liver pathological sections (HE), AST, ALT and LDH levels in model group were significantly higher than those in negative group (P<0.01), the above indexes in positive drug group were significantly lower than those in negative group (P<0.01,P<0.05), and the above indexes in TFSA’s low-dose, medium-dose and high-dose groups were significantly lower than those in model group(P<0.01,P<0.05).The results of CCl4-induced rat hepatocyte injury in vitro showed that AST, ALT, LDH, MDA and NO levels were significantly higher and SOD level was lower in model group than those in negative group (P<0.01), AST, ALT, LDH, MDA and NO levels were significantly lower and SOD level was higher in positive drug group than those in model group (P<0.01, P<0.05), and AST, ALT, LDH, MDA and NO levels were significantly lower and SOD level was higher in TFSA ’ s low-dose, medium-dose and high-dose groups were significantly lower than those in model group ( P <0.01, P <0.05 ) .Conclusion TFSA have hepatoprotective effects on CCl4-induced chemical liver injury and BCG+LPS-induced immune liver injury in mice, and rat hepatocyte damage.This study provides experimental data for the development and utilization of Scorzonera austriaca Wild resources and new hepatoprotective medicines.