Differential proteomic analysis of human genetic prion disease patients in frontal lobe tissues / 中华实验和临床病毒学杂志

Objective@#To search for biomarkers for human familial prion disease.@*Methods@#Two-dimensional differential gel electrophoresis (2D-DIGE) proteomic analysis has been performed in frontal lobe tissues of 3 patients suffering from human familial prion disease (PrP) and 3 age-and sex-matched patients suffering from sudden death due to heart failure without neurological disease.@*Results@#The maps revealed 14 polypeptide chains differentially modulated in the PrP samples, among those, 7 could be identified upon digestion and MALDI-TOF/MS analysis, of which 6 appeared to be up-regulated, 1 being down-regulated.@*Conclusions@#We highlight Galectin-1(Gal-1), ryanodine receptor 2 (RyR2), ubiquitin, Rab-interacting lysosomes protein-like protein 1 (RILPL-1) profillin 2 (PFN2), in the differential map. These proteins are related to neurogenesis, the clearance of misfolded proteins, stasis of calium channel, myoclonus and so on. These proteins are potential biomarkers or targets for treatment of prion disease.

Differential proteomic analysis of liver tissues between male and female C57BL / 6J mice by 2D-DIGE / 中国比较医学杂志

Objective To identify the differential proteomic expressions between the liver tissues of male and female mice, and investigate the mechanisms underlying gender differences in liver diseases. Methods Two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry ( MALDI-TOF-MS) were used to identify the differentially expressed proteins in the liver tissues of male and female C57BL/6J mice. The differentially expressed proteins were validated by Western blot and further analyzed by bioinformatics, including Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG). Results Among the auto-detected 1767 protein spots by 2D-DIGE, 325 protein spots were differentially expressed (|ratio|≥1. 5, P< 0. 05) between the liver tissues of male and female mice, in which 78 spots were randomly selected for MALDI-TOF-MS identification and finally 48 distinct proteins were obtained. Compared with females, 14 and 34 proteins were up-or down-regulated in males, respectively. Among them, 6 differentially expressed proteins were validated by Western blot which confirmed the reliability of 2D-DIGE results. GO analysis showed that the differentially expressed proteins in the liver tissues of male and female mice are associated to various cellular component, molecular function and biological process. 6 pathways were significantly different between the liver tissues of males and females depending on KEGG analysis. Conclusions The proteomic data and related analysis of the liver tissues of C57BL/6J mice offer crucial clues for elucidating the underlying mechanisms of different gender effects on liver diseases.

Differential proteomic analysis of liver tissues between male and female C57BL / 6J mice by 2D-DIGE / 中国比较医学杂志

Objective To identify the differential proteomic expressions between the liver tissues of male and female mice, and investigate the mechanisms underlying gender differences in liver diseases. Methods Two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry ( MALDI-TOF-MS) were used to identify the differentially expressed proteins in the liver tissues of male and female C57BL/6J mice. The differentially expressed proteins were validated by Western blot and further analyzed by bioinformatics, including Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG). Results Among the auto-detected 1767 protein spots by 2D-DIGE, 325 protein spots were differentially expressed (|ratio|≥1. 5, P< 0. 05) between the liver tissues of male and female mice, in which 78 spots were randomly selected for MALDI-TOF-MS identification and finally 48 distinct proteins were obtained. Compared with females, 14 and 34 proteins were up-or down-regulated in males, respectively. Among them, 6 differentially expressed proteins were validated by Western blot which confirmed the reliability of 2D-DIGE results. GO analysis showed that the differentially expressed proteins in the liver tissues of male and female mice are associated to various cellular component, molecular function and biological process. 6 pathways were significantly different between the liver tissues of males and females depending on KEGG analysis. Conclusions The proteomic data and related analysis of the liver tissues of C57BL/6J mice offer crucial clues for elucidating the underlying mechanisms of different gender effects on liver diseases.

Effects of differential expression saliva proteins on OLP patients with Huashi Xingyu Qingre decoction treatment / 实用医学杂志

Objective To analyze the effect of Huashi Xingyu Qingre decoction therapy through identification of the differentially expressed saliva proteins of oral lichen planus. Method The saliva of OLP patients before and after treatment were collected. Total saliva proteins were extracted. The differentially expressed saliva proteins were screened by two-dimensional fluorescence difference gel electrophoresis and identified by liquid chromatography-mass spectrometry. The differentially expressed proteins were analyzed by Western-blot. Results Six differentially expressed proteins were identified as salivary amylase, serum albumin, IgM, carbonic anhydrase VI, zinc-α2- glycoprote and sIgA. The expression level of serum albumin, IgM, carbonic anhydrase VI and zinc-α2-glycoprotein after treatment were lower than that before. However, the expression level of sIgA was higher. The differences were statistically significant. Conclusions Some differentially expressed saliva proteins of OLP before and after Huashi Xingyu Qingre decoction therapy are characterized, and they may play a vital part in the occurrence and development of OLP.

Differentially expressed serum proteins in oral lichen planus patients before and after treated with Huashi Xingyu Qingre Decoction / 医学研究生学报

Objective Oral lichen planus ( OLP) is a common complaint in the oral mucosa , for which there is no definite therapy hitherto.This article aimed to investigate the possible effect of Huashi Xingyu Qingre Decoction (HXQD) on OLP. Methods This study included 30 randomly selected cases of OLP treated with HXQD .Fasting venous blood and total serum protein were obtained before and after medication for screening and identification of OLP-related differential proteins by two-dimensional fluorescence differ-ence gel electrophoresis (2D DIGE) and liquid chromatography-mass spectrometry (LC-MS), followed by Western blot validation . Results Haptoglobin , antithrombin Ⅲ, C1 complement , and vitamin D binding protein were differentially expressed in the serum of the OLP patients before and after treated with HXQD .Compared with the baseline , the expression of haptoglobin was significantly in-creased (103.086 ±27.536 vs 159.704 ±24.228, P<0.05) while that of antithrombin Ⅲ remarkably decreased after treatment (150.00 ±54.04 vs 98.00 ±28.04, P<0.05). Conclusion Haptoglobin, antithrombin Ⅲ, C1 complement, and vitamin D binding protein are differentially expressed in the serum of OLP patients before and after HXQD medication , which may be associated with the development and progression of OLP .

Proteomic analysis of hepatocellular carcinoma HepG2 cells treated with platycodin D / 中国天然药物

Platycodin D (PD), a triterpenoid saponin isolated from Platycodonis Radix, is a famous Chinese herbal medicine that has been shown to have anti-proliferative effects in several cancer cell lines. The aim of this study was to determine the changes in cellular proteins after the treatment of hepatocellular carcinoma HepG2 cells with PD using proteomics approaches. The cell viability was determined using the MTT assay. The proteome was analyzed by two-dimensional difference gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot analysis was used to confirm the expression of changed proteins. Our results showed that PD inhibited the proliferation of HepG2 cells in concentration- and time-dependent manners. Sixteen proteins were identified to be up-regulated in PD-treated HepG2 cells, including ATP5H, OXCT1, KRT9, CCDC40, ERP29, RCN1, ZNF175, HNRNPH1, HSP27, PA2G4, PHB, BANF1, TPM3, ECH1, LGALS1, and MYL6. Three proteins (i.e., RPS12, EMG1, and KRT1) decreased in HepG2 cells after treatment with PD. The changes in HSP27 and PHB were further confirmed by Western blotting. In conclusion, our results shed new lights on the mechanisms of action for the anti-cancer activity of PD.

Screening of differentialy expressed saliva proteins from oral lichen planus patients by two-dimensional fluo-rescence difference gel electrophoresis and mass spectrometry / 实用口腔医学杂志

Objective:To identify differentially expressed saliva proteins of oral lichen planus(OLP)patients by two-dimensional fluo-rescence difference electrophoresis(2-D DIGE)and mass spectrometry(MS).Methods:3 pairs of saliva samples from OLP patients and matched healthy adults were collected.Saliva proteins were separated by 2-D DIGE and identified by liquid chromatography-mass spectrometry(LC-MS).Results:SDS-PAGE examination showed that the electrophoresis bands were clear and protein loss was rare. Protein dots were highly reproducible by 2-D DIGE.In average,the abundance of (31 7 ±71 )saliva protein spots were found in OLP pa-tients.4 highly reproducible spots were identified to be secretory IgA1 ,zincα-2-glycoprotein,salivary amylase and serum albumin by LC-MS and they were at higher level in OLP patients than those in the healthy controls.Conclusion:Secretory IgA1 ,zincα-2-glyco-protein,salivary amylase and serum albumin are highly expressed in the saliva of OLP patients,and may be related to the occurrence and development of oral lichen planus.

Proteomics analysis of human brain glial cell proteome by 2D gel.

INTRODUCTION: Proteomics is increasingly employed in both neurological and oncological research, and applied widely in every area of neuroscience research including brain cancer. Astrocytomas are the most common glioma and can occur in most parts of the brain and occasionally in the spinal cord. Patients with high‑grade astrocytomas have a life expectancy of <1 year even after surgery, chemotherapy, and radiotherapy. MATERIALS AND METHODS: We extracted proteins from tumors and normal brain tissues and then evaluated the protein purity by Bradford test and spectrophotometry method. In this study, we separated proteins by the two‑dimensional (2DG) gel electrophoresis method, and the spots were analyzed and compared using statistical data. RESULTS: On each analytical 2D gel, an average of 800 spots was observed. In this study, 164 spots exhibited up‑regulation of expression level, whereas the remaining 179 spots decreased in astrocytoma tumor relative to normal tissue. Results demonstrate that functional clustering and principal component analysis (PCA) has considerable merits in aiding the interpretation of proteomic data. Proteomics is a powerful tool in identifying multiple proteins that are altered following a neuropharmacological intervention in a disease of the central nervous system (CNS). CONCLUSION: 2‑D gel and cluster analysis have important roles in the diagnostic management of astrocytoma patients, providing insight into tumor biology. The application of proteomics to CNS research has invariably been very successful in yielding large amounts of data.

Early diabetic nephropathy plasma proteomic analysis / 重庆医学

Objective To explore the sensitive blood plasma molecular markers in diabetic nephropathy (DN) .Methods Two-di-mensional fluorescence difference gel electrophoresis (2D-DIGE) was used to analyze early DN patients plasma (n=8) and normal plasma ,proteins that showed differential expression of a 1 .5 fold change were analyzed by MALDI-TOF/TOF mass spectrometry . Results 2D-DIGE maps of plasma proteins in patients with DN and normal plasma were established successfully .We validated 13 differentially expressed proteins detected by 2D-DIGE ,including complement component C3、complement component C4、apolipopro-tein E ,etc .Conclusion Proteomic analysis can objectively revealed the differences of protein expression between the two kinds of blood plasma .The 13 proteins might be the potential plasma molecular markers for the clinical diagnosis of diabetic nephropathy .

Recombinant E. coli expressing Vitreoscilla haemoglobin prefers aerobic metabolism under microaerobic conditions: A proteome-level study.

Vitreoscilla haemoglobin (VHb) expression in heterologous host was shown to enhance growth and oxygen utilization capabilities under oxygen-limited conditions. The exact mechanism by which VHb enhances the oxygen utilization under oxygen-limiting conditions is still unknown. In order to understand the role of VHb in promoting oxygen utilization, changes in the total protein profile of E. coli expressing the vgb gene under its native promoter was analysed. Two-dimensional difference gel electrophoresis (2D DIGE) was employed to quantify the differentially expressed proteins under oxygen-limiting conditions. Overexpression of proteins involved in aerobic metabolic pathways and suppression of proteins involved in non-oxidative metabolic pathways shown in this study indicates that the cells expressing VHb prefer aerobic metabolic pathways even under oxygen limitation. Under these conditions, the expression levels of proteins involved in central metabolic pathways, cellular adaptation and cell division were also found to be altered. These results imply that Vitreoscilla haemoglobin expression alters aerobic metabolism specifically, in addition to altering proteins involved in other pathways, the significance of which is not clear as of now.

Quantitative Proteomics Analysis of LCM Purified Stroma of Nasopharyngeal Carcinoma and Normal Nasopharyngeal Mucosa / 生物化学与生物物理进展

The mechanism of how stroma plays an important role in tumor carcinogenesis is now a hotspot. To delineate the features of stromal protein between nasopharyngeal carcinoma (NPC) and normal nasopharyngeal mucosa(NNM), laser capture microdissection (LCM) was performed to purify stromal cells from the NPC and NNM, respectively. The protein expressed profiles of the stroma of NPC and NNM were compared using fluorescent two-dimensional difference gel electrophoresis (2D-DIGE) and 34 differential protein spots between tumor stroma and normal stroma were chosen to be identified by mass spectrometry (MS). A total of 20 differential proteins were identified, and three differential proteins (CapG, L-plastin and S100A9) were selectively further validated by Western blotting and immunohistochemical analysis to confirm the results of 2D-DIGE. 2D-DIGE patterns of the stroma of NPC and NNM were established for the first time, the results suggested that differentially expressed proteins in the stroma of NPC and NNM may be useful for understanding the relationship between NPC cells and their surrounding microenvironment. Further studying of these proteins will be helpful to elucidate the mechanisms of NPC carcinogenesis and provide new thoughts on therapy of NPC through stroma.