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.
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
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.
Objective To find a new specific marker that can be used to early diagnose hepatic fibrosis by detecting the change of serum protein in patients with hepatic fibrosis. Methods This research adopted 50 SD rats (25 males and 25 females), and from which 6 rats were selected randomly (3 males and 3 females) as control group, last 44 rates were divided into four groups according to four pathological stages as hepatic fibrosis model group (experimental group). Distinct proteins in serum were detected by surface-enhanced laser desorption/ionization-time of flight-mass spectra (SELDI-TOF-MS). Radioimmunoassay was used to measure four parameters of hepatic fibrosis which were hyaluronidase (HA), precollagen Ⅲ (PCⅢ), laminin (LN) and collagen Ⅳ (Ⅳ-C). Results Distinct proteins in serum were detected in 8 cases of stage Ⅰ of hapatic fibrosis, 5 cases of stage Ⅱ, 5 cases of stage Ⅲ, 6 cases of stage Ⅳ, and 5 cases of control by SELDI-TOF-MS. Three protein peaks were found (M/Z: 4 203, 4 658, and 7 400). The peaks of M/Z 4 658 and 4 700 proteins were obviously increased in the stage Ⅰ of hepatic fibrosis （Plt;0.05）, while the changes of hepatic fibrosis four parameters appeared in stage Ⅳ of hepatic fibrosis. Conclusion This method shows great potential for early diagnosing of hepatic fibrosis and finding better biomarkers to hepatic fibrosis.
ObjectiveTo analyze the differences in proteins between aneurysm/dissection patients and healthy subjects, and subsequently figure out differential proteins related to medial degeneration of aortic aneurysm/dissection.MethodsAortic wall samples were collected from 6 male aortic aneurysm patients (an aortic aneurysm group, mean age 56.50±8.19 years), 6 male aortic dissection patients (an aortic dissection group, mean age 54.17±6.68 years) and 6 male healthy subjects (a normal group, mean age 40.50±9.31 years) between December 2019 and May 2020 in West China Hospital of Sichuan University. Quantitative proteomics was performed using tandem mass tag (TMT) techniques, followed by gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis.ResultsA total of 63 differential proteins were obtained both in the aortic aneurysm group and the aortic dissection group compared with the normal group, with 30 up-regulating and 33 down-regulating. The differential proteins were involved in multiple biological processes and clusted on peroxisome proliferators-activated receptor (PPAR) signaling pathway, extracellular matrix-receptor interaction signaling pathway and complement and coagulation cascades signaling pathway.ConclusionThe identified proteins may help to demonstrate new molecular mechanisms related to medial degeneration of aortic aneurysm/dissection.
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.
ObjectiveTo characterize proteomic profile in aqueous humor of patients with pathologic myopia (PM) using quantitative proteomic analysis, which may provide new clues to understand the mechanisms and possible treatments of PM.MethodsA cross-sectional study. From January 2019 to August 2019, aqueous humor samples (32 cataract patients) were collected for quantitative proteomic analysis using liquid chromatography tandem mass spectrometry at Tianjin Medical University Eye Hospital. There were 11 males and 21 females. They were 58-76 years old with an average age of 68.41±6.09 years old. Sixteen patients with PM were regarded as PM group, 16 patients without myopia were regarded as the control group. The aqueous humor samples (100-150 μl ) were collected from all patients before cataract surgery. Using protein quantification and non-labeled liquid chromatography tandem mass spectrometry analysis, differentially expressed proteins were obtained. Five different proteins were randomly selected for ELISA verification. The differentially expressed proteins were further analyzed by gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes, which were validated using ELISA in the other twenty samples of each group.ResultsA total of 583 proteins were identified and 101 proteins were found to be differentially expressed, including 63 up-regulated proteins and 38 down-regulated proteins. ELISA verification results showed that the expression trend of the 5 differentially expressed proteins between the PM group and the control group was consistent with the results of Label-free quantitative proteomics analysis. The main classifications of these differentially expressed proteins were protein-binding activity modulator, defense/immunity protein, protein modifying enzyme, metabolite interconversion enzyme, extracellular matrix protein, transfer/carrier protein and so on. The bioinformatics analysis suggested that PM was closely associated with inflammation and immune interactions, and remodeling of extracellular matrix.ConclusionsCompared with the control group, the protein expression profile of PM patients' aqueous humor specimens has obvious changes. These differences indicate that PM is closely related to inflammation and immune interaction and extracellular matrix remodeling.
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.