With the advance of drug development and research techniques, the drug metabolic processes and mechanism can be more deeply achieved. As the drug metabolism and pharmacokinetics process are mediated by drug metabolizing enzymes and transporters, study of drug metabolizing enzymes and transporters has become an important part for drug development. The traditional immunoassays with low sensitivity and poor specificity can not reflect the accurate expression level of drug metabolizing enzymes and transporters. We now give a brief review on the quantitative study of drug metabolizing enzymes and transporters by mass spectrometry-based proteomic approach.

Objective To explore the effect of hesperidin on cardiac function and ventricular remodeling following myocardial infarction (MI) in mice.Methods Ligation of left anterior descending (LAD) was operated to establish MI model.Forty-two C57BL/6 mice were randomly (random number) divided into control and MI group;and 24 h after LAD ligation,mice in MI group were further divided into MI control and hesperetin group.Eight weeks later,cardiac function and structure changes were determined by the methods of hemodynamic measurement and echocardiography.HE staining was used to measure crosssectional area (CSA) of atrial myocytes,and PSR staining was applied for observe collagen deposition and calculation of collagen volume fraction (CVF).Real-time PCR was used to detect the mRNA expressions of cardiac hypertrophy markers (ANP,BNP and β-MHC) and cardiac fibrosis markers (Collagen Ⅰ,Collagen Ⅲ and CTGF).The contents of superoxide anion and hydroxy radical were detected by colorimetric method.Results Compared with control group,left ventricular posterior wall thickness (LVPWT) and interventricular septum thickness (IVST) were increased to be thicker,left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) were significantly lower,and ± dp/dtmax was remarkably reduced in MI control group (P ＜ 0.05).Compared with MI control group,hesperetin could increaseLVFS [(29.48±3.87)％ vs.(20.69±3.99) ％],LVEF [(46.40±1.68)％ vs.(30.51± 1.17) ％] and ±dp/dtmax [(3 344.33 ±269.57) mmHg/S vs.(2 205.19 ±224.17) mmHg/S;(2250.40±218.35) mmHg/S vs.(-1 566.91 ±217.37) mmHg/S];but could reduce LVPWT [(2.29±0.05) mm vs.(2.85±0.10)mm]andIVST[(1.44±0.09) mm vs.(1.89±0.06) mm].Compared with control group,CSA and CVF were significantly increased in MI control mice.However,hesperetin could reduce CSA and CVF.Compared with control group,the mRNA expressions of cardiac hypertrophy and cardiac fibrosis markers were significantly increased in MI control mice;but hesperetin could significantly inhibit the mRNA expressions of cardiac hypertrophy and cardiac fibrosis markers.Additionally,hesperetin could significantly reduce the contents of superoxide anion and hydroxy radical.Conclusion Hesperetin intervention can inhibit ventricular structure change,and improve hemodynamics and cardiac function after acute myocardial infarction via inhibiting the production of reactive oxygen species (ROS).

The human serum proteome is closely associated with the state of the body. Endogenous peptides derived from proteolytic enzymes cleaving on serum proteins are widely studied due to their potential application in disease-specific marker discovery. However, the reproducibility of peptidome analysis of endogenous peptides is significantly influenced by the proteolytic enzymes within body fluids, thereby limiting the clinical use of the endogenous peptides. We comprehensively investigated the N and C terminus of endogenous peptides using peptidomics. The cleavage site patterns of the N and C terminus and adjacent sites from all the identified endogenous peptides were highly conserved under different sample preparation conditions, including long-term incubation at 37°C and pretreatment with repeated freeze-thaw cycles. Furthermore, a distinguishable cleavage site pattern was obtained when a different disease serum was analyzed. The conserved cleavage site pattern derived from proteolytic enzymes holds potential in highly specific disease diagnosis.
Carcinoma, Hepatocellular ; blood ; diagnosis ; Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Humans ; Liver Neoplasms ; blood ; diagnosis ; Mass Spectrometry ; Nanotechnology ; Peptide Hydrolases ; metabolism ; Peptides ; blood ; Protein Structure, Tertiary ; Proteome ; analysis ; Proteomics ; Silicon Dioxide ; chemistry ; Time Factors

Purpose To investigate the effect and molecu-lar mechanism underlying SOX9 during esophageal squamous cell carcinoma(ESCC)cells.Methods Immunohistochemistry(IHC)was used to detect the expression of SOX9 in ESCC tis-sues and adjacent normal tissues.The correlation of SOX9 ex-pression with clinicopathological features and prognosis of ESCC was analyzed.The differentially expressed genes in Eca109-Vec-tor and Eca109-SOX9 cells were detected by Affymetrix miRNA array.qRT-PCR was used to determine the differential gene in TE-1 and TE-1-siSOX9 cells.The relationship between SOX9 and active/unphosphorylated β-catenin levels was detected by Western blot.The effects of Wnt inhibitor LGK974 on the prolif-eration of ESCC cells were detected by CCK-8.Results SOX9 was highly expressed in ESCC(4.58±3.04)as compared with that in adjacent normal tissues(1.56±2.08,P＜0.001).SOX9 was related to histopathological grade and invasion depth(P＜0.05).Kaplan-Meier analysis indicated high SOX9 expres-sion in ESCC was significantly correlated with shorter overall sur-vival(P＜0.05).Transcriptome profiling and qRT-PCR analysis suggested that SOX9 contributed to the regulation of AXIN2,FZD7 and WNT5A.Overexpression of SOX9 in Eca109 cells in-creased active/unphosphorylated β-catenin levels,whereas silen-cing SOX9 caused a decrease.Significant attenuation of cell pro-liferation was observed at various concentrations of LGK974 with-out affecting SOX9 expression on SOX9-expressing cell lines.As expected,this inhibitory effect was not obvious in Eca109-Vector cells when compared with Eca109-SOX9 cells treated with the same concentration of LGK974.Conclusion SOX9 is highly ex-pressed in ESCC and SOX9-mediated Wnt/β-catenin signal path-way activation at least partially contributes to the SOX9-induced ESCC growth.These findings suggest that SOX9 is a promising prognostic marker and therapeutic target for ESCC.

Objective To investigate the expression,synergistic relationship and clinical significance of alcohol de-hydrogenase(ADH1A)and vascular endothelial growth factor-A(VEGFA)in hepatocellular carcinoma(HCC).Methods The expression and correlation of ADH1A and VEGFA in HCC and adjacent normal tissues were ana-lyzed by GEPIA.TCGA and GSEA were used to analyze the pathway of ADH1A in HCC.The clinical and patho-logical data of 84 patients with HCC were collected,and 54 patients with paracancer normal tissue samples were se-lected as controls to analyze the correlation between ADH1A and VEGFA and clinicopathological parameters of HCC.Immunohistochemistry was used to detect the protein expression of ADH1A and VEGFA in cases and con-trols,and the correlation between the expression of ADH1A and VEGFA and the clinical progression and prognosis of patients with HCC was analyzed based on clinical pathological parameters and Kaplan-Meier.Results Bioinfor-matics analysis found that ADH1A was low-expressed in HCC and VEGFA was highly expressed in HCC,and there was a negative correlation between the two(P<0.001);immunohistochemical detection results showed that the expression of ADH1A in HCC tissue was lower than that in normal tissue adjacent to cancer(P<0.01)while the expression rate of VEGFA in HCC tissue was significantly higher than that of normal tissue adjacent to cancer(P<0.01);The recurrence rate of vascular thrombus and HCC patients in HCC group with high expression of ADH1A was lower(P<0.05).The proportion of tumor diameter>5 cm,high TNM stage,microsatellite and G2-G3 dif-ferentiation in HCC tissues in VEGFA high expression group was higher(P<0.05).Kaplan-Meier survival analy-sis showed that patients with high ADH1A expression and low VEGFA expression had a higher five-year survival rate.Conclusion Low expression of ADH1A and high expression of VEGFA in tumor tissues of patients with HCC indicate tumor progression and can be used as one of the prognostic evaluation indicators for patients with HCC.