Objective To select relatively specific biomarkers in serum from lung adenocarcinoma patients using surface-enhanced laser desorption ionization time of flight mass spectrometry （SELDI-TOF-MS） Protein Chip technology, and study the follow-up results of postoperative serum proteomic patterns. Methods Serum samples from 71 lung adenocarcinoma patients. 71 healthy volunteers with matched gender, age and history of smoking were analyzed by using weak cation exchange 2（WCX2） Protein Chip to select potentially biomarkers. Seventy-one patients were followed-up till 9 months after surgery. Compare the serum proteomic patterns 3,6 and 9 months after surgery. Results Five highly expressed potential biomarkers were identified with the relative molecular weights of 4 047.79, 4 203. 99, 4 959. 81, 5 329. 30 and 7 760. 12 Da. The postoperative serum proteomic patterns changed among individuals, and correlated with patients＇ clinical stage. Conclusions SELDI-TOF-MS Protein Chip technology is a quick, easy, convenient, and high-throughout analyzing method capable of selecting relatively specific, potential biomarkers from the serum of lung adenocarcinoma patients and may have attractive clinical value.
Objective Toa nalyzed ifferentialp roteine xpressiono fc holangiocarcinomai np eripheralb loodb yproteomics technology, and to investigate the significance of proteomics technology in early diagnosis of bile ductmalignancy.M ethods Serum proteinf rom 58p atientsw ithc holangiocarcinomaa nd5 8c ontrols( 20p atientsw ithcholecystolithiasis and 38 healthy people) were detected by surface enhanced laser desorption/ionization-time offlight-mass spectrometry (SELDI-TOF-MS). Ciphergen protein chip software was used to identify proteinic spectra.R esults Comparedw itht hes pectrao fs erum proteini nc ontrolg roup,t herew ere1 0d ifferentiallye xpressedproteins in bile duct carcinoma group, among which three proteins with relative molecular masses of 5. 900 X 10’，9.08 0X 1 0’a nd1 1.86 3X 1 0’w ereu p-regulated( Plt;1 0-’)ands evenp roteinsw ithr elativem olecularm asseso f6.9 59X 1 0’，14.0 00X 1 0’，14.1 29X 1 0’，14.3 02X 1 0’，17.5 57X 10’，17.6 90X 1 0’a nd2 8.5 52X 1 0’w ered ownregulated(Plt; 10-’)。The average concentration of protein with the relative molecular mass of 11. 863 X 10’ incholangiocarcinoma group was eight times more than that in controls group. At the stage I of cholangiocarcinoma,thee xpressiono fp roteinp ointw itha r elativem olecularm asso f5 .90 0X1 0’w ass ignificantlyh ighert hant hosep atientsat the stage III and stage fV (Plt;10-’)，while there were no statistical difference of expression between diffeent clinical stages for the other 9 proteins points. And there were no significant expression differences of the above10 proteins between the patients with and without jaundice following cholangiocarcinoma. Instead, another threeproteinsw ithr elativem olecularm asseso f7 .25 5X 1 03，12.36 4X 1 0’a nd1 5.8 73X 1 03w ered etectedt oh aved ifferentproteine xpressions.A nda llo fth em showedh ighe xpressionsin j aundiceg roup( Plt;10-5).C onclusion Thereare remarkable differences of the expressions of serum proteins in peripheral blood in patients with cholangiocarcinoma.T hep roteinp ointw itha r elativem olecularm asso f1 1.86 3X 1 0’m ayb ea p otentialb iomarkerfo re arlyd iagnosisof cholangiocarcinoma
ObjectiveTo explore the possible active mechanism of the basic fibroblast growth factor (bFGF) long circulation l iposome (LCL) (bFGF+LCL) on spinal cord traction injury in rats at the level of proteomics. MethodsTwenty Sprague Dawly rats were randomly divided into groups A and B, 10 rats in each group. The models of spinal cord traction injury was established at T12-L3 spines. The rats were not treated in group A, and the rats were treated with bFGF+LCL (20μg/ kg) in group B. At 3 weeks after operation, the rats were sacrificed for harvesting T13-L2 spinal tissue specimens. The protein was extracted and quantified in the spinal tissue firstly. The proteins from spinal tissue were separated by two-dimensional gel electrophoresis and identified by mass spectrometry. The different expression profiling was established in each group, and the differentially expressed protein was determined by comparing the level of each spot with gel imaging software and manually. The proteins were identified by nano ultra-high performance liquid chromatography-electrospray tandem mass spectrometry (NanoUPLC-ESI-MS/MS), and the proteins were classified. ResultsThe differentially expressed protein spots were found in 2 groups. Compared with group A, 4 spots were up-regulated and 6 were down-regulated in group B. NanoUPLC-ESI-MS/MS results showed that 18 significant proteins were identified in 26 differentially expressed proteins, including 4 apoptosis-related proteins, 3 nerve signal transduction related proteins, 7 proteins involved in metabolism, 1 unknown function protein, and 3 unnamed proteins. ConclusionThe differentially expressed proteins are found in spinal cord traction injury of rats treated with bFGF+LCL. bFGF+LCL can affect the proteins expression in rats with spinal cord traction injury. The possible active mechanism is that it has protective and repair effects on injured spinal cord by nerve signal transduction, and regulation of nerve cells apoptosis and metabolism.
ObjectiveTo observe the proteomic changes in vitreous fluid samples from patients with rhegmatogenous retinal detachment combined with choroidal detachment (RRDCD). MethodsA prospective cross-sectional clinical study. Vitreous fluid samples were collected from 35 patients with RRDCD (RRDCD group) and 40 patients with rhegmatogenous retinal detachment (RRD group) who were diagnosed at Wuhan Aier Eye Hospital between November 2021 and December 2023. Prior to vitrectomy, 0.3-0.5 ml of vitreous fluid was collected from the affected eyes. Differentially expressed proteins were analyzed using Data-Independent Acquisition (DIA). Three of these proteins were randomly selected for validation using enzyme-linked immunosorbent assay (ELISA). Bioinformatics analyses, including gene ontology functional enrichment and kyoto encyclopedia of genes and genomes pathway enrichment, were performed to explore the functions of the differentially expressed proteins. ResultsSignificant differences were observed between the RRDCD and RRD groups in intraocular pressure (t=－12.795), the number of retinal tears (t=4.601), the extent of retinal detachment (χ2=39.642), axial length (t=0.840), postoperative proliferative vitreoretinopathy incidence (χ2=4.730), single-surgery reattachment rate (χ2=7.717), and best-corrected visual acuity (t=7.033) at 6 months postoperatively (P＜0.05). A total of 237 differentially expressed proteins were identified between the RRDCD and RRD groups, with 63 upregulated and 174 downregulated. These proteins were involved in pathways such as extracellular matrix-receptor interaction, complement activation, coagulation, and lysosomal pathways. ELISA validation results showed that the expression trends of the three selected proteins in the RRDCD and RRD groups were consistent with the DIA proteomic analysis. Compared to the RRD group, proteins such as fibrin, coagulation factors, cathepsins, and trypsin inhibitors were significantly upregulated in the RRDCD group.ConclusionsThe protein expression profile in vitreous fluid samples from RRDCD patients show significant alterations compared to the RRD group. These differential changes suggest that RRDCD is closely associated with complement and coagulation cascade activation, lysosomal pathways, and extracellular matrix remodeling.
ObjectiveTo summarize the overall diagnostic accuracy of serum proteomic assay for pulmonary tuberculosis through a Meta-analysis.MethodsStudies regarding the diagnostic utility of serum proteomic assay for pulmonary tuberculosis were searched in Scopus, PubMed, Wanfang, China National Knowledge Infrastructure, and CQVIP. The methodical quality was evaluated by Quality Assessment for Studies of Diagnostic Accuracy Studies-2 tool. The pooled sensitivity, specificity, positive/negative likelihood ratios, and diagnostic odds ratio were calculated. Summary receiver operating characteristic curve was generated and the area under the curve was calculated.ResultsThere were 10 articles with 2 433 patients included in this study, containing 1 191 cases and 1 242 controls. The pooled sensitivity, specificity, positive/negative likehood ratios, and diagnostic odds ratio were 0.86, 0.88, 6.72, 0.17, and 46.84, respectively. The area under the curve was 0.93.ConclusionSerum proteomic assay plays a role in diagnosing pulmonary tuberculosis, and proteomic assay represents a novel and useful method for diagnosing pulmonary tuberculosis.
Abstract： Objective To study the molecular mechanism of pathologic states related to cardiopulmonary bypass (CPB) and screen the differential proteins from the serum before and after CPB in the open heart surgery patients. Methods By the twodimensional gel electrophoresis (2DE), we took the blood samples from each of the sixteen open heart surgery patients 30 minutes before CPB, 1 hour after CPB, and 24 hours after CPB. The protein spots were analyzed by the PDQuest image analysis software and the differential protein spots were identified by matrixassisted laser desorption/ionizationtime of flightmass spectrometry (MALDITOF-MS). Then, enzymelinked immunosorbent assay (ELISA) was used to determine the expression level of serum amyloid A protein (SAA) in the serum of healthy people and the enrolled patients before and after CPB. Results Through 2DE in combination with massspectrometry, 7 proteins altered in expression were identified, including SAA, haptoglobin (HPT), leucinerich alpha2-glycoprotein (A2GL), hemoglobin subunit beta (HBB), serine/threonineprotein phosphatase 2A -regulatory subunit B″ subunit gamma (P2R3C), transthyretin (TTHY), and T-complex protein 11-like protein2 (T11L2). ELISA analysis showed that SAA levels in healthy people and the open heart surgery patients 30 minutes before CPB were not statistically different (t=-1.955, P=0.056), while the SAA level rose from 54.47±48.32 μg/ml 30 min before CPB to 1 017.78±189.92 μg/ml 24 hours after CPB in the serum of open heart surgery patients． Conclusion The results of this pilot study illustrate that SAA, HPT, A2GL, HBB, P2R3C, TTHY and T11L2 may be the molecule markers of pathologic state related to CPB. Acute phase reaction happens intensively after CPB in human body.
ObjectiveTo establish two-dimensional electrophoresis profiles with high resolution and reproducibility from subcellular immortalized human endocervical cell (H8) and cervical cancer cell (Caski), and to identify the differential expressions of subcellular proteins (cytoplasmic, membranous and nuclear proteins). MethodsH8 cells and Caski cells were incubated, and subcellular proteins of H8 cells and Caski cells were extracted and separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). Then the selected differential protein spots were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and SWISS-PROT database. ResultsWe obtained well-resolved, reproducible 2-DE patterns; 6 differentially expressed cytoplasmic proteins and 3 differentially expressed membranous proteins and 9 differentially expressed nuclear proteins were defined in 2-DE gels. ConclusionSubcellular proteins of cervical precancerous lesion and cervical cancer are separated and analyzed by means of 2-DE and MALDI-TOF-MS/MS. There are significant differences between H8 cells and Caski cells. These data may be valuable for research of cervical precancerous lesion and cervical cancer, or as diagnostic markers and therapeutic targets for cervical cancer.
Objective To identify proteins that have expressed in human eyes from adults and two-month old infants by proteomics approach, so as to build a two-dimensional gel electrophoresis (two-DE) reference map for human retina. The difference of proteomics between the retinas of adults and two-month old infants are also studied. Methods Human retina tissues were collected from donor eyes (nine adults and two infants). Proteins were separated by two-DE. The gels were analyzed by image software. Protein spots were excised from the gels and detected by matrix assisted laser desorption ionization time off light mass spectrometry (MALDI-TOF-MS). Results A total of 1179 spots and 1295 spots were detected respectively on two-DE gels of Coomassie-stained adults and two-month old infants retina, of which 1039 spots were matched in the position. Five spots up-regulated were successfully identified. Human serum albumin and 4 guanylate kinase 1 (GUK1) were identified in adult retina. beta;2-tubulin, transaldolase1 and alpha A-crystallin were identified in infant retina. Conclusion The two-DE reference map for retina proteomics is successfully established. This study provides an evidence of changes in retinal protein levels between adults and infants and biochemical pathways for future studies of human retina development.
ObjectiveTo analyze the protein expression changes in the retina of non-arteritic anterior ischemic optic neuropathy (NAION) in rats.MethodsThe rat NAION (rNAION) model was established by Rose Bengal and laser. Twenty Sprague-Dawley rats were randomly divided into 4 groups, the normal control group, the laser control group, the RB injection control group, and the rNAION model group, with 5 rats in each group. The right eye was used as the experimental eye. The retina was dissected at the third day after modeling. Enzyme digestion method was used for sample preparation and data collection was performed in a non-dependent collection mode. The data were quantitatively analyzed by SWATH quantitative mass spectrometry, searching for differential proteins and performing function and pathway analysis.ResultsCompared with the other three control groups, a total of 184 differential proteins were detected in the rNAION group (expression fold greater than 1.5 times and P＜0.05), including 99 up-regulated proteins and 85 down-regulated proteins. The expressions of glial fibrillary acidic protein, guanine nucleotide binding protein 4, laminin 1, 14-3-3γ protein YWHAG were increased. Whereas the expressions of Leucine-rich glioma-inactivated protein 1, secretory carrier-associated membrane protein 5, and Clathrin coat assembly protein AP180 were decreased. The differential proteins are mainly involved in biological processes such as nerve growth, energy metabolism, vesicle-mediated transport, the regulation of synaptic plasticity, apoptosis and inflammation. Pathway enrichment analysis showed that PI3K-Akt signaling pathway and complement and thrombin reaction pathway was related to the disease.ConclusionThe protein expressions of energy metabolism, nerve growth, synaptic vesicle transport and PI3K-Akt signaling pathway can regulate the neuronal regeneration and apoptosis in NAION.
ObjectiveTo study the differentially expressed proteins of recombinant human erythropoietin (r-HuEPO) in hippocampus of Pentetrazol (PTZ) -induced epileptic rats, and to provide a basis for exploring the pathogenesis of epilepsy and seeking new therapeutic targets. Methods Twelve 6~8-week-old Sprague Dawley rats that weighted 230~250 g were randomly divided into two groups: PTZ group, PTZ+ EPO group. The differential proteins of recombinant human EPO in hippocampus of pentylenetetrazole-induced epileptic rats were analyzed and identified by TMT technique based on mass spectrometry.Results 139 differentially expressed protein sites were detected in hippocampal tissues of epileptic rats, of which 55 were up-regulated and 84 down-regulated. Conclusion Recombinant human erythropoietin can inhibit many differentially expressed proteins in the hippocampus of pentaerythraze-induced eclampsia rats by upregulation of Isocitrate dehydrogenase (NADP), Reduced nicotinamide purine dinucleotide phosphate (NADPH), Thioredoxin reductase 2 mitochondrial (TrxR), reduce nerve cell damage.