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ObjectiveHenoch-Schönlein purpura（HSP） is one of the dominant diseases in Mongolian medicine. Qishun Baolier（QSBLE）， as the main prescription for the treatment of HSP， has significant clinical effect， but its mechanism is not yet clear. Baed on this， this study is intended to screen the differentially expressed proteins before and after treatment， and preliminarily explore the molecular mechanism of QSBLE in the treatment of HSP. MethodTaking oneself as the control， 30 HSP patients aged 6-45 years were collected， and QSBLE was taken orally at 12：00 and 24：00， respectively. The dose was adjusted according to age and the course of treatment was one week. The distribution of proteinuria， hematuria and skin purpura of all patients were determined before and after treatment. The serum samples of 10 patients with clinically significant remission after QSBLE treatment were randomly selected for proteomics. Isobaric tags for relative and absolute quantification（iTRAQ） combined with liquid chromatography tandem mass spectrometry（LC-MS/MS） was used to analyze the proteins in serum of HSP patients before and after treatment， and differential proteins were analyzed bioinformatically and the protein-protein interaction（PPI） networks were constructed. ResultA total of 378 proteins were identified from serum， including 18 differentially expressed proteins， of which 15 proteins were up-regulated and 3 proteins were down regulated. Bioinformatics showed that the differential proteins were mainly involved in biological processes such as immune response， immunoglobulin production， phagocytosis， adaptive immune response before and after treatment. Biological processes， pathways and proteins were used to construct the PPI network， the proteins represented by immunoglobulin heavy constant γ1（IGHG1）， immunoglobulin λ-chain 7-43（IGLV7-43）， gelsolin（GSN） and 60 kDa heat shock protein（HSPD1） were involved in biological processes and related pathways such as adaptive immune response， immunoglobulin production， leukocyte-mediated immunity， regulation of stress response， regulation of immune system processes， regulation of trauma response， and these proteins were at the center of the PPI network. ConclusionQSBLE may play a role in the treatment of HSP by regulating the expression of IGHG1， IGLV7-43， GSN， HSPD1 and other key proteins to affect immune-related biological processes.
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OBJECTIVE@#The study explores the effects of electroacupuncture (EA) at the governing vessel (GV) on proteomic changes in the hippocampus of rats with cognitive impairment.@*METHODS@#Healthy male rats were randomly divided into 3 groups: sham, model and EA. Cognitive impairment was induced by left middle cerebral artery occlusion in the model and EA groups. Rats in the EA group were treated with EA at Shenting (GV24) and Baihui (GV20) for 7 d. Neurological deficit was scored using the Longa scale, the learning and memory ability was detected using the Morris water maze (MWM) test, and the proteomic profiling in the hippocampus was analyzed using protein-labeling technology based on the isobaric tag for relative and absolute quantitation (iTRAQ). The Western blot (WB) analysis was used to detect the proteins and validate the results of iTRAQ.@*RESULTS@#Compared with the model group, the neurological deficit score was significantly reduced, and the escape latency in the MWM test was significantly shortened, while the number of platform crossings increased in the EA group. A total of 2872 proteins were identified by iTRAQ. Differentially expressed proteins (DEPs) were identified between different groups: 92 proteins were upregulated and 103 were downregulated in the model group compared with the sham group, while 142 proteins were upregulated and 126 were downregulated in the EA group compared with the model group. Most of the DEPs were involved in oxidative phosphorylation, glycolipid metabolism and synaptic transmission. Furthermore, we also verified 4 DEPs using WB technology. Although the WB results were not exactly the same as the iTRAQ results, the expression trends of the DEPs were consistent. The upregulation of heat-shock protein β1 (Hspb1) was the highest in the EA group compared to the model group.@*CONCLUSION@#EA can effect proteomic changes in the hippocampus of rats with cognitive impairment. Hspb1 may be involved in the molecular mechanism by which acupuncture improves cognitive impairment.
Subject(s)
Rats , Male , Animals , Rats, Sprague-Dawley , Electroacupuncture , Proteomics , Cognitive Dysfunction/therapy , HippocampusABSTRACT
A growing body of evidence has linked the gut microbiota to liver metabolism. The manipulation of intestinal microflora has been considered as a promising avenue to promote liver health. However, the effects of Lactobacillus gasseri LA39, a potential probiotic, on liver metabolism remain unclear. Accumulating studies have investigated the proteomic profile for mining the host biological events affected by microbes, and used the germ-free (GF) mouse model to evaluate host-microbe interaction. Here, we explored the effects of L. gasseri LA39 gavage on the protein expression profiles of the liver of GF mice. Our results showed that a total of 128 proteins were upregulated, whereas a total of 123 proteins were downregulated by treatment with L. gasseri LA39. Further bioinformatics analyses suggested that the primary bile acid (BA) biosynthesis pathway in the liver was activated by L. gasseri LA39. Three differentially expressed proteins (cytochrome P450 family 27 subfamily A member 1 (CYP27A1), cytochrome P450 family 7 subfamily B member 1 (CYP7B1), and cytochrome P450 family 8 subfamily B member 1 (CYP8B1)) involved in the primary BA biosynthesis pathway were further validated by western blot assay. In addition, targeted metabolomic analyses demonstrated that serum and fecal β-muricholic acid (a primary BA), dehydrolithocholic acid (a secondary BA), and glycolithocholic acid-3-sulfate (a secondary BA) were significantly increased by L. gasseri LA39. Thus, our data revealed that L. gasseri LA39 activates the hepatic primary BA biosynthesis and promotes the intestinal secondary BA biotransformation. Based on these findings, we suggest that L. gasseri LA39 confers an important function in the gut‒liver axis through regulating BA metabolism.
Subject(s)
Mice , Animals , Bile Acids and Salts/metabolism , Lactobacillus gasseri , Proteomics , Liver/metabolism , BiotransformationABSTRACT
Objective To compare the differentially expressed proteins between cypermethrin-resistant and -sensitive Culex pipiens pallens, so as to unravel the mechanism underlying the resistance to cypermethrin in Cx. p. pallens. Methods A quantitative proteomic analysis was performed among cypermethrin-sensitive and -resistant isolates of Cx. p. pallens using isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with liquid chromatography with tandem mass spectrometry (LC-MS/MS). Results A total of 164 differentially expressed proteins were identified between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens, including 54 up-regulated proteins and 110 down-regulated proteins. A large number of cuticular proteins, larval cuticular proteins, pupal cuticular proteins and cuticular structural constituent proteins, which are associated with cytoskeletal structure and components, were differentially expressed between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens. Thirteen proteins, which were involved in energy production and conversion, translation, ribosomal structure and biogenesis, lipid transport and metabolism, post-translational modification, protein turnover, chaperones, cytoskeleton and intracellular transportation, were validated to be differentially expressed between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens, which may serve as potential markers of cypermethrin resistance. Conclusion Multiple insecticide resistance mechanisms contribute to the resistance to cypermethrin in Cx. p. pallens, including cuticular resistance and metabolic resistance, and the cuticular protein genes and cytochrome P450 enzymes may play an important role in the resistance of Cx. p. pallens to cypermethrin.
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In the past decade, relative proteomic quantification using isobaric labeling technology has developed into a key tool for comparing the expression of proteins in biological samples. Although its multiplexing capacity and flexibility make this a valuable technology for addressing various biological questions, its quantitative accuracy and precision still pose significant challenges to the reliability of its quantification results. Here, we give a detailed overview of the different kinds of isobaric mass tags and the advantages and disadvantages of the isobaric labeling method. We also discuss which precautions should be taken at each step of the isobaric labeling workflow, to obtain reliable quantification results in large-scale quantitative proteomics experiments. In the last section, we discuss the broad applications of the isobaric labeling technology in biological and clinical studies, with an emphasis on thermal proteome profiling and proteogenomics.
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Proteome/metabolism , Proteomics/methods , Reproducibility of Results , Tandem Mass Spectrometry/methodsABSTRACT
The present study aimed to explore the mechanism of the sweating of Dipsacus asper on content changes of triterpene sa-ponins by detecting the total triterpene saponins and the index component asperosaponin Ⅵ in the crude and sweated D. asper, and analyzing the differentially expressed proteins by isobaric tags for relative and absolute quantification(iTRAQ) combined with LC-MS/MS. After sweating, the content of total triterpene saponins decreased manifestly, while that of asperosaponin Ⅵ increased significantly. As revealed by the iTRAQ-LC-MS/MS analysis, 140 proteins with significant differential expression were figured out, with 50 up-regulated and 90 down-regulated. GO analysis indicated a variety of hydrolases, oxido-reductases, and transferases in the differential proteins. The results of activity test on two differentially expressed oxido-reductases were consistent with those of the iTRAQ-LC-MS/MS analysis. As demonstrated by the analysis of enzymes related to the triterpene saponin biosynthesis pathway, two enzymes(from CYP450 and UGT families, respectively, which are involved in the structural modification of triterpene saponins) were significantly down-regulated after sweating. The findings suggested that sweating of D. asper presumedly regulated triterpene saponins by affecting the expression of downstream CYP450 s and UGTs in the biosynthesis pathway of triterpene saponins of D. asper.
Subject(s)
Humans , Chromatography, Liquid , Dipsacaceae , Saponins , Sweating , Tandem Mass Spectrometry , TriterpenesABSTRACT
Objective: By applying the proteomic method, to analyze differences of protein expressions and signal pathways in veins at arteriovenous fistula in chronic kidney disease stage 5 (CKD5) patients with or without diabetes so as to explore the pathogenesis of high incidence of arteriovenous fistula functional incapacitation in CKD5 patients with diabetes. Methods: The protein expression RAW datasets of vascular access from CKD5 patients with or without diabetes in Proteomexchange Database were screened out and downloaded. Then the identified features were searched from UniProt/SwisssProt human proteins database through the software ProteomeDiscovery (PD). Then, the PD generated a file of quantitative proteins data. The significantly different proteins between two groups were screened out and analyzed by T-test or Adj T-test depending on homogeneity of variance. These significantly different proteins were enriched into different biological pathways through IPA analysis, GO enrichment analysis, and KEGG pathway analysis, which signifies these biological pathways are significantly different. Ultimately, the correlation of proteins in different biological pathway was analyzed by Spearman correlation test. Results: TheiTRAQ labeled proteins RAW datasets comparing the vessels from CKD5 patients with diabetes and without diabetes (PXD010883) were collected at first. After searching identified features again and disposing the data, a total of 120 significantly different proteins including 89 up-regulated proteins and 31 down-regulated proteins were screened out finally. Through GO, KEGG and IPA analyses, there were two significantly different biological pathways. On the one hand, the purine nucleoside monophosphate metabolic change and the ratio of ATP/AMP decrease were reflected in oxidative phosphorylation receding and AMP metabolism enhancing in CKD5 patients with diabetes. On the other hand, the muscle system process which played a vital role in VSM cells receded in CKD5 patients with diabetes. It was specific in the decrease of MYH11, CNN1 and so on. Excitingly, the significantly differential genes in the two pathway had a strong correlation. The results explained why CKD5 patients with diabetes always have cardiovascular disease. Conclusion: The ratio of ATP/AMP reduction caused by diabetes led to muscle function disorder of VSM cells, which explains the reason for the internal fistula functional incapacitation in CKD5 patients with diabetes.
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BACKGROUND: The pellicle, the acellular organic material deposited on the surface of tooth enamel, has been thought to be derived from saliva. In this study, protein compositions of the pellicle, gingival crevicular fluid, and saliva collected from healthy adults were compared to elucidate the origin of pellicle proteins. RESULTS: The pellicle, gingival crevicular fluid, and saliva from the parotid gland or mixed gland were collected; subsequently, protein expression in samples from the respective individual was compared by SDS-PAGE and mass spectrometry. Following SDS-PAGE, proteins in the major bands were identified by mass spectrometry. The band pattern of pellicle proteins appeared different from those of gingival crevicular fluid, or saliva samples. Using mass spectrometry, 13 proteins in these samples were identified. The relative abundance of the proteins was quantitatively analyzed using mass spectrometry coupled with stable isotope labeling and by western blot. Cystatin S and α-amylase detected in pellicle were enriched in saliva samples, but not in gingival crevicular fluid, by western blot, and their abundance ratios were high in saliva and low in gingival crevicular fluid when analyzed by stable isotope labeling. Serotransferrin, however, was found only in the pellicle and gingival crevicular fluid by western blot and its abundance ratio was low in saliva. CONCLUSIONS: Our study revealed that the gingival crevicular fluid appears to contribute to pellicle formation in addition to saliva.
Subject(s)
Humans , Male , Female , Adult , Saliva/chemistry , Proteins/analysis , Gingival Crevicular Fluid/chemistry , Dental Pellicle/chemistry , Mass Spectrometry , Blotting, Western , Electrophoresis, Polyacrylamide GelABSTRACT
Context: Colorectal cancer (CRC) is one of the most common malignancies and one of the leading causes of cancer death worldwide. Establishing early detection methods or markers of CRC is central to improve the survival rate of CRC patients. Nowadays, new molecular tools have been developed to acquire further knowledge on tumor progression. Aims: Comparative proteomics analysis of Vietnamese colorectal carcinoma in different stages was performed to gain an insight into the molecular events taking place in CRC and metastasis. Subjects and Methods: In this study, the comparative protein expression analysis of ten paired CRC and its corresponding noncancerous tissue samples was performed using the combination of isobaric tags for relative and absolute quantitation labeling and mass spectrometry (MS). The data obtained were further analyzed with Ingenuity Pathways Analysis (IPA) system. Results: Based on the MS/MS spectra analyzed by ProteinPilot software, 684 proteins were identified, out of which 215 were observed to be differentially expressed in at least 1 sample pair. Individual protein expression and variation have been identified for each patient. IPA system demonstrated cytoskeletal signaling as the top-ranked functional pathway network associated with the oncogenic function. Conclusions: Our study supplemented the understanding about proteome of Vietnamese CRC patients and identified statistically protein expression differences among samples assisting in finding effective biomarkers for CRC diagnostics
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Objective@#To screen the deferentially expressed proteins in hippocampus and striatum in rat models of diabetes mellitus and normal SD rats, and to elucidate the effects of hyperglycemia on central nervous system.@*Methods@#SD rats were randomly divided into normal group(n=10)and hyperglycemia group(n=15); rat models of diabetes were induced by the high-fat and high-sugar diet combined with single intraperitoneal injection of streptozotocin. The hippocampus and striatum tissues from hyperglycemia rat and normal SD rat(control group) were selected as specimens. Isobaric tags for relative and absolute quantification(iTRAQ) technique and 2-dimensional liquid chromatography-tandem mass spectrometry(2D-LC-MS/MS) were used for identifying the deferentially expressed proteins. The deferentially expressed proteins were analyzed by bio-informatics for searching candidate proteins.Finally, the candidate proteins were verified by Western blotting.@*Results@#A total of 347 proteins showed significantly different expressions, which included 139 up-regulated proteins and 289 down-regulated proteins. These proteins included metabolic glutamate receptor 2, ferritin, leucine repeat protein and complex 2, which are related to Parkinson′s disease. KEEN search indicated that the pathway of dopaminergic synaptic, complement, gap junction, autophagic and Fcgamma R-mediated phagocytosis were involved with these significant differentially expressed proteins. The protein interaction network showed that ferrin, aldehyde/ketone reductase(ARK), Ca2+ -binding protein(CaBP), and protein tyrosine phosphatase(PTP) were located at the crossed nodes. The expression levels of ferrin, ARK, PTP, and CaBP, which were verified by Western blotting, and the results were consistent with those of the quantitative mass spectrometry(P<0.05).@*Conclusion@#Deferentially expressed proteins in hippocampus striatum of diabetic rats can be effectively screened by iTRAQ technique combined with 2D-LC-MS/MS. It may provide new clues for the mechanism of neurodegenerative disease.
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Objective To compare the difference of protein expression between the post-overwintering stage and the diapauses preparation stage in Culex pipiens pallens, so as to reveal the mechanisms underlying the overwintering diapause of Cx. pipienspallens. Methods A quantitative proteomic analysis was performed in Cx. pipiens pallens before and after overwintering diapause by using isobaric tags for relative and absolute quantification (iTRAQ) labeling. Results A total of 244 differentially expressed proteins were identified in Cx. pipiens pallens before and after overwintering diapause, including 126 up-regulated proteins and 118 down-regulated proteins. iTRAQ-based quantitative proteomic analysis revealed that these differentially expressed proteins were linked to function and energy production and conversion, lipid metabolism, remodeling of cytoskeleton, carbohydrate metabolism, protein transport, molecular chaperones, stress tolerance and metabolic enzymes. Conclusions This is the first study to identify the overwintering diapause-related proteins in Cx. pipiens pallens using proteomics tools, which reveals KEGG pathways and GO terms associated with the overwintering diapauses of Cx. pipiens pallens. Our findings provide additional understandings pertaining to the mechanisms underlying the overwintering diapauses of Cx. pipiens pallens.
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Objective To compare the difference of protein expression between the post-overwintering stage and the diapauses preparation stage in Culex pipiens pallens, so as to reveal the mechanisms underlying the overwintering diapause of Cx. pipienspallens. Methods A quantitative proteomic analysis was performed in Cx. pipiens pallens before and after overwintering diapause by using isobaric tags for relative and absolute quantification (iTRAQ) labeling. Results A total of 244 differentially expressed proteins were identified in Cx. pipiens pallens before and after overwintering diapause, including 126 up-regulated proteins and 118 down-regulated proteins. iTRAQ-based quantitative proteomic analysis revealed that these differentially expressed proteins were linked to function and energy production and conversion, lipid metabolism, remodeling of cytoskeleton, carbohydrate metabolism, protein transport, molecular chaperones, stress tolerance and metabolic enzymes. Conclusions This is the first study to identify the overwintering diapause-related proteins in Cx. pipiens pallens using proteomics tools, which reveals KEGG pathways and GO terms associated with the overwintering diapauses of Cx. pipiens pallens. Our findings provide additional understandings pertaining to the mechanisms underlying the overwintering diapauses of Cx. pipiens pallens.
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Objective@#To compare the differentially expressed proteins in mice with kidney-yang deficiency and those with kidney-yin deficiency induced by hydrocortisone, and explore the similar and different material bases of male infertility caused by the two types of kidney deficiency.@*METHODS@#Thirty Kunming mice were equally randomized into a normal control, a kidney-yang deficiency and a kidney-yin deficiency group. The animals of the normal control group were injected intraperitoneally with normal saline at 0.2 ml qd for 7 days, while those of the latter two groups with hydrocortisone at 25 mg/kg/d for 10 days and 50 mg/kg/d for 7 days, respectively, for establishment of kidney-yang deficiency and kidney-yin deficiency models. Then the pathological changes in the testicular tissue of the mice were observed by HE staining and the differentially expressed proteins were compared among different groups using isobaric tags for relative and absolute quantitation (iTRAQ) and the bioinformatics method.@*RESULTS@#Sod1 was found to be a reproduction-related node protein differentially expressed in the testis tissues of the two types of kidney-deficiency mice, more highly expressed in the kidney-yin than in the kidney-yang deficiency group (P < 0.05). Five reproduction-associated node proteins were co-expressed in the testes of the two groups of kidney-deficiency mice, with significantly up-regulated expression of Rps28 and down-regulated expressions of Rpl11, Rplp2, Svs2 and Svs3a (P < 0.01).@*CONCLUSIONS@#Sod1 may be one of the key material bases for the differentiation of male infertility caused by kidney-yang deficiency from that induced by kidney-yin deficiency, while Rps28, Rpl11, Rplp2, Svs2 and Svs3a may be the common material bases of male infertility caused by the two types of kidney deficiency.
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Background and Objectives: Bupivacaine is available as a racemic mixture of dextro and levobupivacaine. Many studies show that dextrobupivacaine has greater cardiovascular and central nervous system toxicity than levobupiva-caine. The objectives of the present study were to compare the effects of racemic Bupivacaine + Fentanyl and Levo-bupivacaine + Fentanyl on the complete regression of motor block, onset time to reach T10level sensory block, dura-tion of T10level sensory block, onset time of motor block, duration of sensory block. Materials and Method: The study was conducted in 100 patients undergoing transurethral resection of prostate operation, who received either 1.75 ml Bupivacaine (0.5%) + 25 μg Fentanyl (Gr A) or 1.75 ml Levobupivacaine (0.5%) + 25 μg Fentanyl (Gr B) in-trathecally. Results: Time to complete regression of motor block, onset time toT10level sensory block were signifi-cantly prolonged in Gr A compared to Gr B. The onset time of motor block was significantly shorter in Gr A compared to Gr B. There was no statistically significant difference between the two groups in respect to the duration of T10level sensory block, duration of sensory block. Conclusion: Intrathecal Levobupivacaine + Fentanyl used in the present study can be considered as a suitable alternative to Bupivacaine + Fentanyl for spinal anaesthesia in elective TURP surgery
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Differential proteomics analysis of Sika deer antlers at rapid growth stage (60 d) and ossification stage ( 90 d) was performed by isobaric tags for relative and absolute quantitation ( iTRAQ ) , ultra high performance liquid chromatography and mass spectrometry technologies. A total of 127 differential proteins were identified. Compared with the ossification stage, 80 differential proteins were significantly up-regulated and 47 differential proteins were significantly down-regulated at the rapid growth stage. These differential proteins were mainly distributed in the regions of extracellular matrix, nucleosome, haptoglobin-hemoglobin complex, actin filament, endoplasmic reticulum-Golgi intermediate compartment, endoplasmic reticulum lumen, and endometrium, etc. The up-regulated differential proteins were mainly involved in the regulations of oxygen transport in the blood, nerve growth and regeneration, cartilage and bone development and ATP synthesis compared with ossification stage, and the down-regulated differential proteins were mainly involved in the endochondral ossification process. The changes of protein expression at different growth stages were closely related to antler rapid growth and ossification. Therefore, the results of this study provided a basic data for discovering the molecular mechanisms of antler rapid growth and ossification, and it was of great significance for further study of the pharmacological basis and clinical application of antlers.
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Helicobacter pylori infection is related to the development of gastric diseases. Our previous studies showed that high thioredoxin-1 (Trx1) expression in H. pylori can promote gastric carcinogenesis. To explore the underlying molecular mechanisms, we performed an isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic analysis of stomach tissues from Mongolian gerbil infected with H. pylori expressing high and low Trx1. Differences in the profiles of the expressed proteins were analyzed by bioinformatics and verified using Western blot analysis. We found three candidate proteins, 14-3-3α/β, glutathione-S-transferase (GST), and heat shock protein 70 (HSP70), in high Trx1 tissues compared with low Trx1 tissues and concluded that cellular stress and redox activity-related proteins were involved in the pathogenesis of gastric cancer associated with H. pylori Trx1.
Subject(s)
Animals , 14-3-3 Proteins/physiology , Computational Biology , Gerbillinae , Glutathione Transferase/physiology , HSP70 Heat-Shock Proteins/physiology , Helicobacter Infections/complications , Helicobacter pylori , Oxidation-Reduction , Stomach Neoplasms/etiology , Stress, Physiological , Thioredoxins/physiologyABSTRACT
Objective To analyze the protein expression profile of HeLa cells transfected with pORF5 gene of Chlamydia trachomatis. Methods A lentiviral expression vector containing pORF5 gene was constructed. The lentiviral expression vector and helper plasmids were co-transfected into 293T cells to construct the recombinant lentivirus, which was used to infect HeLa cells. HeLa cells transfected with pORF5 gene and control HeLa cells were sorted out by flow cytometry. The isobaric tags for relative and absolute quantitation ( iTRAQ) approach combined with nano-liquid chromatography-tandem mass spec-trometry ( NanoLC-MS/MS) analysis was performed to understand protein expression profiles and to iden-tify and quantify the differentially expressed proteins in the pORF5-transfected HeLa cells ( pORF5-Hela) and the control HeLa cells. Quantitative real-time PCR ( qRT-PCR ) and Western blot analysis were performed to detect the expression of some proteins at mRNA and protein levels, respectively. Results HeLa cell line stably transfected with pORF5 gene and control HeLa cell line were constructed successful-ly. Totally 314 proteins were differentially expressed between the pORF5-HeLa and control HeLa cells, 159 of which showed increased expression and the other 155 showed decreased expression in pORF5-HeLa cells. The differentially expressed proteins were involved in many processes, such as metabolic process, immune response, biological adhesion and so on. Results of qRT-PCR showed that the expression of HIST1H1C(histone H1. 2C), HBA1(hemoglobin subunit alpha), PARK7(parkinson disease protein 7), HMGB1(high mobility group protein B1) and HMGB2 at mRNA level in pORF5-HeLa cells were up-regulated, while the expression of CLIC1 ( chloride intracellular channel protein 1 ) , KRT7 ( typeⅡ cy-toskeletal 7), SFN(14-3-3 protein sigma) and CDKN2A(cyclin-dependent kinase inhibitor 2A) were down-regulated. Western blot analysis confirmed the enhanced expression of HMGB1 and PRAK7 at pro-tein level. The results of qRT-PCR and Western blot analysis were consistent with proteomic data. Con-clusion Expression profiles for differentially expressed proteins between pORF5-HeLa and control HeLa cells were established successfully. The differentially expressed proteins regulated by pORF5 gene were found to be related to cell metabolism, proliferation, adhesion and so on, suggesting that pORF5 might promote the growth and proliferation of Ct by regulating protein expression and biological behavior of host cells.
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Objective To determine the expression of serum proteins in pancreatic cancer patients based on mass spectrometry to screen differential proteins and to find potential molecular biomarkers to prediagnosis of pancreatic cancer. Methods The technique of isobaric tags for relative and absolute quantitation (iTRAQ) and two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) were used to analyze the serum proteins in 15 pancreatic cancer patients and 10 healthy controls. Screening for serum differential proteins was performed via retrieving Panther, Mascot and Scaffold databases. Ingenuity Pathway Analysis was used to detect the correlation between proteins. Results The expression level of 442 proteins was quantified, and 76 proteins were found to be differentially expressed which changed corresponding biological process. The up-regulation of DNA repair protein 50 (RAD50) and down-regulation of transforming growth factor β1 (TGFβ1) and apoptosis protease-activating factor 1 (APAF1) which were correlated with cell growth and apoptosis were found in pancreatic cancer. A closely interacted network was formed between these three differential proteins and other molecules, which could effect metabolism in pancreatic cancer patients. Conclusion Differential expression of serum proteins screened by iTRAQ in pancreatic cancer patients may be the potential markers of pancreatic cancer, which can provide molecular basis for early diagnosis of pancreatic cancer.
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Objective To screen for serum protein differentially expressed between women whose fetuses had congenital heart defects(CHD) and women who had normal fetuses. Methods Serum samples were collected from pregnant women whose fetuses had CHD and those whose fe?tuses had no CHD,including a CHD group of 40 women and a control group of 10 women. The CHD group included 4 subgroups as follows:tetralo?gy of Fallot,ventricular septal defects,persistent truncus arteriosus,and a mixture of relatively rare types of CHD(n=10 each). Samples in the same group were pooled to obtain equal amounts of proteins ,and the iTRAQ proteomic approach was used to identify and quantify the proteins dif?ferentially expressed among these groups. Results We successfully identified 606 proteins,among which 47 showed at least a 1.5?fold difference between the CHD and control groups. Among the 47 proteins,23 and 24 were upregulated and downregulated,respectively. Conclusion Several proteins associated with CHD could be identified by using the iTRAQ proteomic approach ,and various proteins were involved in the pathogenesis of CHD in this study.
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Objective@#To investigate the effect of aerobic exercise on the spermatogenic function of male rats and screen out differentially expressed proteins related to spermatonesis-regulation by proteomic analysis.@*METHODS@#We randomly divided 24 SD male rats into groups A (non-exercise control), B (exercise), and C (weight-bearing exercise), those in the latter two groups made to swim for 60 minutes a day and those in group C bearing a load 3% of the body weight, both 6 times a week for 9 weeks. At 24 hours after the last exercise, we obtained the sperm count, measured the levels of such serum reproductive hormones as testosterone (T), luteotrophic hormone (LH), follicle-stimulating hormone (FSH), and gonadotrophin-releasing hormone (GnRH), and employed isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis of the testicular tissue.@*RESULTS@#Compared with group A, group C showed significant increases in sperm concentration (［2.12 ± 0.43］ vs ［3.54 ± 0.52］ ×10⁶/ml, P 0.05) and FSH (［20.49 ± 2.44］ vs ［22.29 ± 2.31］ IU/L, P >0.05). No significant changes were observed in sperm concentration or reproductive hormone levels in group B as compared with A. Group B exhibited obviously more mature sperm and cell layers in the seminiferous epithelium than group A. A total of 47 differentially expressed proteins were identified, of which 37 were up-regulated and the other 10 down-regulated. In addition, another 5 significantly differentially expressed proteins closely related to reproductive function were identified, including up-regulated Anx A1, GPX3, Rimbp3, and Dpy19l2 and down-regulated CYP17. Enrichment analysis showed that the differentially expressed proteins were mainly involved in extracellular matrix-receptor interaction, protein digestion and absorption, and focal adhesion pathways.@*CONCLUSIONS@#Proper-intensity exercise can improve the spermatogenic function of rats. Aerobic exercise promotes spermatogenesis mainly by up-regulating the expressions of the proteins related to the production and differentiation of spermatozoa.