Comparative proteomic analysis of Yersinia pestis 201 and 201△pCD1 strains / 军事医学

Objective To explore the potential pathogenesis of Yersinia pestis and provide new clues for vaccine development through comparative proteomic analysis of wild-type and pCD1 cured strain of Yersinia pestis 201.Methods Differentially expressed proteins at 26℃ and 37℃ were separated and identified using two-dimensional electrophoresis coupled with mass spectrometry .Results A total of 24 differently expressed proteins were successfully identified from the samples of bacteria grown at 26℃ and 25 proteins at 37℃.Among these, 7 proteins were encoded by pCD 1 plasmid. Conclusion Through comparative proteomic research, we have found that the abundance of several proteins can be dramatically changed when the large plasmid pCD 1 is missing,suggesting that the plasmid can regulate the expression of many genes located in the chromosome .

A preliminary analysis of the secretome of aggregated dermal papilla cells / 中华皮肤科杂志

Objective To study the expression ot secreted proteins in aggregated dermal papilla cells (DPCs).Methods DPCs were isolated from human scalp tissue and subjected to primary culture and subculture.Aggregated and non-aggregated DPCs served as the subject of this study.Secreted proteins were prepared from these cells and subjected to two-dimensional polyacrylamide gel electrophoresis.Differentially expressed proteins were screened by the PDQuest image analysis software.Protein spots were digested and identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry,and finally analyzed using the National Center for Biotechnology Information (NCBI) non-redundant (Nr) protein database.Results Two-dimensional electrophoresis maps with good repeatability and high resolution were established.Image analysis of 2-D gels revealed that the average number of detected protein spots was 1 134 ± 52 and 1 078 ± 36 in aggregated and nonaggregated DPCs respectively,and the majority of these protein spots were matched between aggregated and nonaggregated DPCs.Twenty-eight protein spots showed more than 5-fold difference between the two groups of cells,and 10 proteins were preliminarily identified as differentially expressed proteins by peptide-mass fingerprinting.Of these differentially expressed proteins,8 proteins including Rhogdi 1,filamin A,cystatin C,fibronectin,cyclophilin A,procollagen C proteinase enhancer 1,tissue inhibitor of metalloproteinase and tissue inhibitor of metalloproteinase-2 were up-regulated,and 2 proteins including neuropolypeptide h3 and matrix metalloproteinase-3/tissue inhibitor of metalloproteinase-1 complex were down-regulated in aggregated DPCs compared with non-aggregated DPCs.Conclusions Differentially expressed proteins between aggregated and non-aggregated DPCs are mainly implicated in cell signaling pathway,cellular proliferation and differentiation,extracellular matrix synthesis and degradation,and so on.

Comparison of two protein preparation methods by two-dimensional gel electrophoresis / 军事医学

Objective To compare the separation effects of protein samples extracted by two different methods with two -dimensional gel electrophoresis (2-DE) .Methods Ultrasonic disruption and glass-beads grounding were used to prepare protein samples of Gram-negative bacteria , Gram-positive bacteria and animal tissues .The actual results of the two sample preparation methods were compared by 2-DE.Results The 2-DE maps of samples extracted by the two methods were obtained.Conclusion The 2-DE maps of glass-beads grounding samples are better than those of ultrasonic disruption thanks to their lower backgrounds , which are beneficial for further image analyses .

Functions of gene phoN1 in Shigella flexneri 2a:preliminary study / 军事医学

Objective To explore the function of gene phoN1 in Shigella flexneri.Method Using the λ-Red recombi-nant system, phoN1was knocked out from S.flexneri 2a strain 301.Comparative proteomics was performed to analyze the differences between mutant and wild-type strains in protein expression profiles .Sereny tests and competitive infection assays were carried out to compare the virulence of mutant and wild-type strains .Results The deletion mutant of phoN1 was suc-cessfully constructed .No significant difference between the two strains was found in the comparative proteomics analyses . The function of gene phoN1 might be unrelated to the invasion ability of S.flexneri according to the results of Sereny tests and competitive infection assays .Conclusion Gene phoN1 might be of no use for the in vitro survival and host cell invasion of S.flexneri.

2D map of proteins from human renal stone matrix and evaluation of their effect on oxalate induced renal tubular epithelial cell injury

Purpose Proteins constitute a major portion of the organic matrix of human calcium oxalate (CaOx) renal stones and the matrix is considered to be important in stone formation and growth. The present study evaluates the effect of these proteins on oxalate injured renal epithelial cells accompanied by a 2D map of these proteins. Materials and Methods Proteins were isolated from the matrix of kidney stones containing CaOx as the major constituent using EGTA as a demineralizing agent. The effect of more than 3kDa proteins from matrix of human renal (calcium oxalate) CaOx stones was investigated on oxalate induced cell injury of MDCK renal tubular epithelial cells. A 2D map of >3kDa proteins was also generated followed by protein identification using MALDI-TOF MS. Results The >3kDa proteins enhanced the injury caused by oxalate on MDCK cells. Also, the 2D map of proteins having MW more than 3kDa suggested the abundance of proteins in the matrix of renal stone. Conclusion Studies indicate that the mixture of >3kDa proteins in the matrix of human renal stones acts as promoter of calcium oxalate crystal nucleation and growth as it augments the renal epithelial cell injury induced by oxalate. The effect of promoters masks the inhibitors in the protein mixture thereby leading to enhanced renal cell injury. 2D map throws light on the nature of proteins present in the kidney stones. .

Screening and identifying oxaliplatin-resistance-associated proteins in colorectal cancer cell lines / 肿瘤

Objective: To screen and identify oxaliplatin-resistance-associated proteins in CRC (colorectal cancer) cell lines using proteomics technologies in order to find new biomarkers for individual therapy of CRC. Methods: Oxaliplatin-resistant human CRC cell line HT-29/L-OHP (oxaliplatin) was established. The total proteins in HT-29 and HT-29/L-OHP cells were extracted. The differentially expressed proteins between HT-29 and HT-29/L-OHP cells were screened and identified using 2-DE (two-dimensional gel electrophoresis) and MALDI-TOF-MS (matrix assisted laser desorption-ionization time-of-flight tandem mass spectrometry). Some proteins obtained were validated by Western blotting. Results: The 2-DE maps of total proteins in HT-29 and HT-29/L-OHP cells were established. Of the 38 protein spots identified as differentially expressed proteins (over two-fold, P < 0.05) between HT-29 and HT-29/L-OHP cells, 37 protein spots were positively identified by MALDI-TOF-MS (17 proteins were up-regulated and 20 proteins were down-regulated as compared with the parental HT-29 cells). The result of Western blotting showed that the PCBP1 [poly (C)-binding protein-1] and TUBB2A (tubulin beta 2A ) proteins were up-regulated while ANXA3 (annexin A3) and STIP1 (stress-induced-phosphoprotein 1) proteins were down-regulated in HT-29/L-OHP cells. The result of Western blotting was consistent with that of proteomics. Conclusion: There were 37 oxaliplatin-resistance-associated proteins in CRC identified in this study which may provide useful evidence in further research on mechanism of oxaliplatin-resistance in CRC. Copyright © 2013 by TUMOR.

Screening for psoriasis-associated proteins by serological proteome analysis / 中华皮肤科杂志

Objective To screen for differentially expressed proteins in sera from patients with common types of psoriasis,and to identify plasma protein markers for psoriasis.Methods Serum samples were collected from 6 patients with progressive psoriasis vulgaris,5 patients with erythroderma psoriaticum,and 6 healthy human controls,and then pooled into 3 pools:psoriasis vulgaris pool,erythroderma psoriaticum pool and control pool.After removal of high-abundance albumin and IgG,the pooled samples were analyzed by two-dimensional electrophoresis (2-DE).An electrophoretic gel image analysis software was used to locate differentially expressed protein spots followed by peptide mass fingerprinting with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS).The National Center for Biotechnology Information (NCBI) protein databases were then searched for the identification of differentially expressed proteins.Results All the three pooled serum samples were well seperated by 2-DE.As the gel image analysis software showed,there were 33 protein spots differentially expressed between the patients with psoriasis vulgaris and the healthy controls,17 between the patients with erythroderma psoriaticum and the healthy controls,and 26 between the patients with psoriasis vulgaris and those with erythroderma psoriaticum.Finally,14 proteins were identified as differentially expressed proteins.The patients with psoriasis vulgaris showed higher expression of complement component 3,interleukin-16,vitamin D-binding protein and α1-antitrypsin compared with the healthy controls; the patients with erythroderma psoriaticum showed increased expression of complement component 3,complement component H,α1-antitrypsin,hemopexin and haptoglobin,but decreased expression of serum amyloid protein compared with the healthy controls,as well as enhanced expression of α1-antitrypsin,complement component H,complement component 4 and haptoglobin compared with those with psoriasis vulgaris.Conclusion Differences exist in serum protein profiles between patients with psoriasis vulgaris and erythroderma psoriaticum,and healthy human controls.

Screening biomarkers in bone marrow supernatant of multiple myeloma by 2-DE and MALDI-TOF-MS / 中华检验医学杂志

Objective To analyze the differentially expressed proteins in bone marrow supernatants of multiple myeloma patients by using 2-DE and MALDI-TOF-MS,and search for the special protein markers for studying the mechanism of the development and diagnosis or differential diagnosis of multiple myeloma.Methods The bone marrow supernatant samples of fourteen multiple myeloma patients,five other hematologic malignancies and five normal controls were collected.After removing albumin and IgG,the proteins in the supernatants were separated by 2-DE.Three groups images were analyzed and compared by Imagemnster 2D platinum 5.0 analysis software.Differentially expressed proteins were selected if the protein spots intensity showed more than 3 fold increase or decrease among different groups.The identities of the differentially expressed proteins with good repeatibility were determined by PMF based on by MALDI-TOF-MS or MALDI-TOF-MS/MS and NCBInr database search.Results 2-DE maps of bone marrow supernatants of the three groups could be analyzed and compared by image analysis of software.Forty-seven and fifty-eight differentially expressed protein spots were detected in multiple myeloma samples compared with normal controls and other hematologic malignancies samples,respectively.Forty-one reproducible spots were analyzed and identified by mass spectrum.Compared with other hematologic malignancies and normal controls,five up-expressed proteins and three down-expressed proteins were identified in multiple myeloma samples.They includes immunoglobulin J chain κ and λ light chain,provirus ancestral Gag polyprotein,mature oxy-cope catalytic antibody with hapten for up-expressed proteins,and hemoglobin,haptoglobin Hp2,zinc-alPha-2-glycoprotein for down-expressed proteins.These differentially expressed proteins reflect the features of muhiple myeloma,and relate to the development,progression and therapy of multiple myeloma.Conclusions Eight differentially expressed proteins in bone marrow supernatants of multiple myeloma are identified by using 2-DE and MALDI-TOF-MS.These differentially expressed proteins could be useful in studying the mechanism of the development and progression of multiple myeloma,and in developing diagnosis and differential diagnosis of multiple myeloma.

Two-dimensional electrophoresis analysis for protein profile change in zebrafish alcohol syndrome model / 中华围产医学杂志

Objective To study the putative mechanisms underlying fetal alcohol syndrome by comparative protein-profile analysis between normal and ethanol-treated zebrafish embryo with twodimensional electrophoresis (2-DE).Methods Zebrafish embryos were exposed in 400 mmol/L ethanol at dome stage for 3 hours,and then ethanol-induced abnormalities were observed.Proteomes of zebrafish embryos at early stages including zygote stage,dome stage,shield stage and 5-somite stage,were separated by 2-DE.The subtraction analysis method was applied to eliminate the interference from maternal derived proteins.The ethanol-treated embryos at 5-somite stage was analyzed by 2-DE,and the protein profile was compared with that generated from control embryos at the same stage.The data obtained from 2-DE analysis were verified by in-situ hybridization.Results 400 mmol/L ethanol treatment caused axial malformation (62%) and cyclopia (60%) in zebrafish embryos.The 2-DE analysis showed that the expression of Collagen2al (Col2a1) and TAR DNA binding protein (TDP) was decreased in 12 hours post fertilization (12 hpf) ethanol-treated embryos by 81% and 73%,respectively.The in-situ hybridization also demonstrated that the expression of Col2al in axial mesoderm was reduced by ethanol treatment at the same stage.But for 24 hpf ethanoltreated embryos,the expression of Col2al in axis recovered to a comparable level to that in control embryos,while the structure of neural tube was disrupted severely by ethanol exposure.Conclusions It is suggested that the expressions of Col2al and TDP were disrupted by ethanol during early stage,which might induce the zebrafish developmental abnormalities.The ethanol interference on early expression of Col2al is supposed to be one of the major reasons leading to later abnormalities of axis and neutral tube.

Application of comparative proteomics in colorectal carcinoma / 国际外科学杂志

Comparative proteomies is to compare the differential expression of overall protein profiles in different time and space,to identify differential proteins or protein groups on the expression amount,expression level,and modification status and to further investigate the differential proteins and their function.The aim of this paper is to introduce the application of comparative proteomies in coloreetal carcinoma on the level of cell,tissue,and body fluid.

A proteomics study of patients with corticobasal degeneration / 中华神经医学杂志

Objective To investigate the molecular mechanism of corticobasal degeneration and search the protein markers ofdiagnoseis and treatment of this disease. Methods The brains of subjects died without clinical or pathological involvement of nervous system (n=4) and brains of patients with corticobasal degeneration (n=4) were obtained at autopsy. Tissues samples were cut from the frontal and temporal lobes, and the striatum. Histological changes of the tissue samples were observed by HE staining, Gallyas-Braak silver staining and Tau protein immunohistochemistry. The samples extracted from the frontal lobe were performed dimensional electrophoresis with immobilized pH gradient (IPG)isoelectric focusing electrophoresis as the first dimension and with vertical sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) as the second dimension; the maps were visualized by Coomassie brilliant blue staining and analyzed with ImageMaster 2D Elite software; the protein profiles were in-gel digested and identified by mass spectrometry (MALDI-TOFTOF). Results Significant neuron loss in the temporal lobe of patients with corticobasal degeneration was noted by HE staining, with mass proliferation of gliocyte in the temporal lobe; sphere-like entanglement in the striatum was noted by Gallyas-Braak silver staining and Tau immunohistochemistry. Fourteen protein spots in the brains of patients with corticobasal degeneration were differentially expressed as compared with those in age-matched nondemented control brains; the expression of 9 proteins (cofilin, uracil DNA glycosylase,Cu-Zn superoxide dismutase, isocitrate dehydrogenase subunit, synaptotagmin Ⅰ, thioredoxin peroxidase 1, glial fibrillary acidic protein, P25 alpha and peroxiredoxin 5) in brains of patients with corticobasal degeneration was up-regulated as compared with that in the normal brain tissue (P＜0.05); the expression of 5 proteins (carbonyl reductase [NADPH] 1, ferritin heavy chain, peptidyl-prolyl cis-trans isomerase A,serum albumin precursor and dihydropyrimidinase-related protein 2) in brains of patients with corticobasal degeneration was down-regulated as compared with that in the normal brain tissues (P＜0.05).Conclusion We get a number of related-proteins of corticobasal degeneration. Some proteins are quite useful in discovering the molecular mechanisms of corticobasal degeneration and may be helpful in the ealry diagnosis and treatment of corticobasal degeneration and in the development of new medicine.

Preliminary study of the impact of plasmid pYC on proteome of Yersinia pestis / 中国地方病学杂志

Objective To investigate the role of plasmid pYC on proteome of Yersinia pestis. Methods Two dimensional electrophoresis was performed to strains of Yersinia pestis with and without the pYC plasmid, and differential proteins were identified by mass spectrometry. Results More than 500 protein spots of Yersinia pestis with and without plasmid pYC were recognized,and their protein profiles were generally similar. The chaperone GroEL was highly expressed in strains with plasmid pYC, whereas the protein GroEL was not encoded by plasmid pYC. ConclusionsPlasmid pYC has an impact on proteome of Yersiniapestis. The proteins of pYC-p10 and pYC-p11 encoded by plasmid pYC may regulate the expression of GroEL.

The application of proteomic technology in differentiation and early diagnosis of myelocytic leukemia / 中华检验医学杂志

Objective To figure out the differentially expressed proteins using proteome technology in leukemia cells induced into different lineages and investigate the application value in early screening of leukemia.Methods With induction of ATRA and NSC67657, the differentiation models was constructed using HL60 cells which has the potentiality to be induced into different lineages by different inducers.Then the differentially expressed proteins in the process of differentiation was separated using two-dimensional electrophoresis and identified using MALDI-TOF MS.The expression of 3 proteins FE1A1, TLE1, NME3 were chosen to be verified in myeloid samples of 5 leukemia patients and 1 normal volunteer using RT-PCR and WB.Results WB showed that NME3 was differentially expressed after both granulocytic and monocytic differentiation( Normal A value = 0.227, NSC67657 A value= 0.079, ATRA A value = 0.064, P ＜ 0.01 ).However, only in 4 of 5 tested patients' myeloid samples, the NME3 protein expression were differentially expressed compared to the normal myeloid sample( Normal A value = 0.082,2 acute leumia transferred from chronic granulocytic leumia A value = 0.274,0.269, acute monocytic leukemia A value = 0.297, one patient with chronic granulocytic leukemia A value = 0.258.There was significant difference between normal and leukemia group, P ＜0.05 ).A value was 0.121 for another patents with chronic granulocytic leukemia The NME3 protein expression was not differentially expressed compared to the normal myeloid sample,P ＞0.05.Conclusions It is still a long way to go for proteome technology from basic research to clinical application.However, the identification of NME3 protein related to differentiation in leukemia patients' myeloid samples had set the foundation for the early diagnosis of leukemia using proteome technology.

Protein expression profile of human glomerular mesangial cells under high glucose / 中华肾脏病杂志

Objective To analyze the protein expression profile of human glomerular mesangial cells (HMCs) under high glucose and to characterize molecular functions and biological processes. Methods HMCs were divided into high glucose cultured group (30 mmol/L) and normal glucose cultured group (5 mmol/L). The total proteins were extracted after culture for 48 hours. The total proteins of the two groups were separated using two-dimensional fluorescence difference in gel electrophoresis (2-D DIGE) and analyzed using DeCyder 2-D difference analysis software. The differentially expressed proteins were further identified using in-gel digestion with trypsin, of which peptide extracts were prepared for MALDI-TOF-MS analysis. Protein identifications were searched in the NCBI protein database using the Mascot search engine. Results One hundred and forty-seven protein spots whose expression levels were significantly increased or decreased more than 1.5 folds under high glucose were identified. Ninety-six differentially expression protein spots were analyzed by peptide mass fingerprinting and 37 kinds of proteins were identified. The protein spots of phosphatidylethanolamine binding protein 1 (PEBP-1), granulysin,ATP synthase H + transporting mitochondrial FO complex subunit F2 were observed only in high glucose group. The expression of 24 proteins was up-regulated by high glucose, including eosinophil cationic protein, RGS membrane-interacting proteins 16 (MIR16), peptidyl-prolyl cis-trans isomerase, disks large homolog DLG2, breast cancer 2, early onset (BRCA2), Catechol-O-methyltransferase etc. The expression of 5 proteins was down-regulated by high glucose, including O-GlcNAc transferase-interacting protein 106 000 isoform 1, proteasome beta 6 subunit precursor,NEFA-interacting nuclear protein NIP30 etc. Conclusions Expression of 147 proteins in HMCs alters under high glucose. These proteins are involved in the regulation of cytoskeleton, glucose metabolism, cell division, gene transcription, signal transduction, phosphorylation, cell proliferation,apoptosis etc. In-depth analysis of these differentially expressed proteins' function and crosstalk is expected to provide an important experimental basis for clarifying the pathogenesis of diabetic nephropathy.

Establishment of a "Four-step method" for extraction and separation of Candida albicans total proteins / 第二军医大学学报

Objective: To introduce a "Four-step method" for extraction and separation of Candida albicans total proteins. Methods: Proteins of C. albicans were extracted step-by-step with 4 kinds of solutions with different solubilities. After quantification, the protein samples were separated by isoelectric focusing electrophoresis and then by SDS-PAGE. Results: Proteins with different solubilities were successfully extracted step-by-step from C. albicans and were separated by two-dimensional gel electrophoresis. Conclusion: The "Four-step method" for extraction of C. albicans proteins is an effective approach to study C. albicans membrane proteome and lays a foundation for further investigation of the mechanisms of antifungal agents and drug resistance in C. albicans.

Proteomic identification of chemosensitivity-associated proteins in human colorectal carcinomas / 肿瘤

Objective:The study aims to screen chemosensitivity-associated proteins in colorectal carcinoma tissues by using two-dimensional gel electrophoresis (2-DE) and mass spectrometry,then identify some differentially-expressed proteins. Methods:The patients with advanced colorectal carcinoma were confirmed by clinical diagnosis. Fresh carcinoma specimens were collected by biopsy and preserved in liquid N2. The tissues were classified into two groups: high sensitivity group (HS) and low sensitivity group (LS) based on drug sensitivity test. The total proteins were extracted and separated by 2-DE. The images were composed, compared, and differentially analyzed to identify the proteins with differential expression in HS and LS groups. Then the differentially-expressed protein spots were incised from the gels and digested by trypsin. The peptide mass fingerprintings (PMF) was acquired after matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching in the Mascot database. Two proteins with differential expression were detected by Western blotting.Results:The 2-DE spectrum of HS and LS groups were established. Most protein spots were distributed in the area with pH 4-8 and relative molecular weight of (20-100)×10~3. The average number of the protein spots was 842±23 in HS group and 793±19 in LS group,respectively. The mean matching rate was 90.7%. The number of differentially-expressed dots between HS and LS group was 79.00±13.56. Thirty protein dots were selected for mass spectrum and bioinformatic analysis, and 9 proteins were identified. Conclusion:Colorectal carcinoma with different chemosensitivity had differential protein expression profiles. The differentially expressed proteins may be associated with chemosensitivity and could be used for prediction of chemosensitivity of colorectal carcinoma.

Polymorphisms of COTL1 gene identified by proteomic approach and their association with autoimmune disorders

To select candidate genes, we attempted to comparative analysis of protein levels between rheumatoid arthritis (RA) patients and healthy controls by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF-MS). We identified 17 proteins that showed up- or down-regulated spots in RA patients. We found that coactosin-like1 (COTL1) were highly expressed in RA patients compared with healthy controls. We performed a case-control study to determine whether the COTL1 gene polymorphisms were associated with RA and systemic lupus erythematosus (SLE). The genotype frequency of c.-1124G>T and the allelic frequency of c.484G>A in RA patients, and the genotype frequency of c.484G>A in SLE patients were significantly different from healthy controls (P = 0.009, 0.027, and 0.025, respectively). We also investigated the correlation with the levels of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCP) antibody in RA patients, and anti-nuclear antibodies (ANA) in SLE patients. The c.484G>A polymorphism in RA patients has significant association with the levels of anti-CCP antibody (P = 0.03). Our findings demonstrated that c.-1124G>T and c.484G>A polymorphisms of the COTL1 gene might be associated with the genetic susceptibility of autoimmune disorders.

Polymorphisms of COTL1 gene identified by proteomic approach and their association with autoimmune disorders

To select candidate genes, we attempted to comparative analysis of protein levels between rheumatoid arthritis (RA) patients and healthy controls by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF-MS). We identified 17 proteins that showed up- or down-regulated spots in RA patients. We found that coactosin-like1 (COTL1) were highly expressed in RA patients compared with healthy controls. We performed a case-control study to determine whether the COTL1 gene polymorphisms were associated with RA and systemic lupus erythematosus (SLE). The genotype frequency of c.-1124G>T and the allelic frequency of c.484G>A in RA patients, and the genotype frequency of c.484G>A in SLE patients were significantly different from healthy controls (P = 0.009, 0.027, and 0.025, respectively). We also investigated the correlation with the levels of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCP) antibody in RA patients, and anti-nuclear antibodies (ANA) in SLE patients. The c.484G>A polymorphism in RA patients has significant association with the levels of anti-CCP antibody (P = 0.03). Our findings demonstrated that c.-1124G>T and c.484G>A polymorphisms of the COTL1 gene might be associated with the genetic susceptibility of autoimmune disorders.

Proteomic analysis of plasma from heroin abuser sunder immobilized Ph gradients 4 -7 / 中国药理学与毒理学杂志

AIM To identify differentially expressed proteins in the plasma that may be used as biomarkers for heroin addiction through a two-dimensional (2-D)gel electrophoresis/mass spectrometry approach. METHODS Following removal of albumin and IgG,the plasma from 5 heroin abusers and 5 normal controls was separated by 2-D gel electrophoresis using immobi-lized pH gradients 4 -7 drystrip and PAGE. Gel ima-ges were analyzed using ImageMaster Elit 5.0. Differ-ential proteins were selected and analyzed through tan-dem mass spectrometry. RESULTS Average number for samples was 350±21 protein spots. In them there were 5 spots that differed by more than 1.5 fold be-tween the two groups obtained through image analysis.Through tandem mass spectrometry above spots were identified as fibrinogen γ (increased by 5 fold), hu-man α1-B-glycoprotein (decreased to 1/1.8 of control group), uncleaved α1-antitrypsin (increased by 2.5 fold), chain of transthyretin (decreased to 1/2 of con-trol group ) and ccruloplasmin (increased by 6.6 fold ). CONCLUSION There are differences between heroin abusers and normal controls in the blood plasma proteome. Some proteins may have a role in the damage to central nervous system through heroin abuse. Such proteins may provide novel biomarkers for diagnosis and therapeutic targeting, as well as clues forunderstanding the mechanism of heroin abuse.

Comparative proteomic study at the different stages during initiation and evolution of experimental colorectal carcinoma / 肿瘤

Objective: To research the proteins differentially expressed during evolution of experimental colorectal carcinoma (normal mucosa→adenoma→carcinoma→liver metastasis) so as to find the early diagnostic biomarker of colorectal cancer as well as to understand its pathogenesis mechanism. Methods: Ninety male rats were injected with 1, 2-dimethylhydrazine intraperitoneally and sacrificed at different weeks to establish the experimental colorectal tumor models (from normal mucosa to liver metastasis). These samples at different stages were collected and divided into four groups (normal mucosa group, adenoma group, carcinoma group, and liver metastasis group). The proteins of these 4 groups were extracted to conduct 2-dimensional gel electrophoresis. The differential protein spots were examined by mass spectrometry and analyzed by bioinformatics. Results: Ten differentially expressed proteins were identified by 2-dimensional gel electrophoresis and mass spectrometry including α-enolase, cardiac α-actin (CA), transgelin protein, myosin regulatory light chain smooth muscle isoform (MRLC), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), haptoglobin, disulfideisomerase (DI), creatine kinase mitochondrial (CKm), heat shock protein-8 (HSP-8) and Keratin complex-2 (KC-2). Conclusions: There exist differentially expressed proteins at various stages during the evolution of colorectal carcinoma. These proteins may be the candidate biomarkers for the early diagnosis of colorectal carcinoma. Proteomic technology is an effective way for preliminary identification of the tumor biomarkers.