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Subject(s)
Female , Humans , Male , Pregnancy , Abortion, Spontaneous , Biological Phenomena , Blotting, Western , Chromatin , Embryo Loss , Embryonic Development , Healthy Volunteers , Karyotyping , Masturbation , Metabolism , Oxidation-Reduction , Oxidative Stress , Prospective Studies , Protein Folding , Proteomics , Sexual Abstinence , Spectrum Analysis , Spermatogenesis , Spermatozoa , Two-Dimensional Difference Gel Electrophoresis , World Health OrganizationABSTRACT
Objective To analyze and compare the differentially expressed plasma proteins between patients with stable angina pectoris (SAP) and unstable angina pectoris (UAP), and search for the biomarkers that maybe used for early diagnosis of UAP. Methods Sixty plasma samples were collected respectively from normal controls group (N group), SAP group and UAP group during Jun. 2014 to Apr. 2015 from the Third Affiliated Hospital of Southern Medical University. Ten samples (100μl) of each group were selected randomly to pool into 3 groups severally. After removing high-abundance proteins from plasma, two-dimensional difference gel electrophoresis (DIGE) was used to isolate the total proteins, and then the protein spots with more than 2-fold changes between UAP and SAP were picked up after the differential software analysis. Afterward, the varied proteins were identified by matrix assisted laser desorption ionization-time of flight/time of flight (MALDI-TOF/TOF) mass spectrometry (MS). Finally, 40 plasma samples were collected respectively from N, SAP and UAP group, and the UAP specific differential proteins were selected to be verified by ELISA. Results A total of 10 varied protein spots with more than 2-fold changes in UAP and SAP were found including 9 up-regulated proteins and 1 down-regulated one. MS identification indicated that the up-regulated proteins included fibrinogen gamma chain (FGG), complement C4-B (C4B), immunoglobulin (Ig) kappa chain C region (IGKC) and hemoglobin subunit alpha (HBA1), whereas the down-regulated one was haptoglobin (HP). After comparing the varied proteins with that in N group, 2 specifically UAP-related proteins, IGKC and HP, were detected totally. IGKC was selected to validate by ELISA, and the corresponding results showed that IGKC was increased specifically in UAP plasma (P<0.05) when compared with N and SAP group, which was consistent with DIGE. Conclusion IGKC and HP have been detected as specifically related proteins to UAP, and IGKC might serve as a potential specific biomarker for screening and early diagnosis of UAP.
ABSTRACT
Objective To determine the probability of identification of differential expression of biliary proteins induced by cholangiocarcinoma using 2D-DIGE. Methods Bile was obtained from 12patients with obstructive jaundice (including 6 cases of cholangiocarcinoma and 6 of cholelithiasis).Each sample was labeled with three different CyDyes (y3,Cy5,Cy2) including one internal standard,pooled from all the samples, and separated with 2-D DIGE in triplicate experiments. MALDI-TOF-MS and bioinformatics were adopted to identify and elucidate the significance of differentially expressed proteins in bile induced by cholangiocarcinoma. Results 55 matched protein spots differences in abundance were detected with statistical variance of two groups(Average Volum Ratio ≥1.5, t-test, P＜0. 05). Among these proteins, 13 PMF were obtained by MALDI-TOF-MS analysis. Eight proteins were identified by searching a protein database. Conclusion The differentially displayed proteomes between the pathological bile obtained from benign and malignant obstructive jaundice indicates the potential application of 2D-DIGE to identify the biomarker of cholangiocarcinoma.