Objective To detect the serum protein fingerprint in gastric cancer patients by using the surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and protein chip array technology, screen biomarker candites, build diagnostic models and evaluate its clinical significance. Methods The serum proteomic patterns were detected in 40 patients with gastric cancer, 20 patients with gastric ulcer and 20 healthy blood donors. The diagnostic models were developed and valited by discriminant analysis. Results The peak intensity of differential expression proteins was not found in healthy blood donors, and 1 case was found in patient with gastric ulcer (m/z: 5 910,4 095). The peak intensity of 5 329, 4 095, 5 910, 8 691 and 3 300 (m/z) proteins were significantly higher in 40 gastric cancer patients than those in 20 gastric ulcer patients and 20 healthy blood donors ( P ＜0.05）. Three differential expression proteins were set up a diagnostic model together to diagnose gastric cancer. The diagnostic model made up of the differential expression proteins of 4 095, 5 910 and 8 691 had a sensitivity of 92.5% and a specificity of 97.5% . Conclusion Using SELDI-TOF-MS shows great potential to detect, and screen novel and better biomarkers for gastric cancer.
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.
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 To study the biological activities ofthe nerve regeneration conditioned fluid (NRCF), especially to further separateand identify the protein bands of the relative molecular mass of (232-440)×103. Methods The silicone nerve regeneration chambers were implanted between the cut ends of the sciatic nerve in 6 New Zealand white rabbits (weight, 1.8-2.5 kg). The proteins in NRCF were separated by the native-polycrylamide gel electrophoresis (Native-PAGE), the protein bands of the relative molecular mass of (232-440)×103 were analyzed by the Shotgun technique, liquid chromatography, and mass spectrometry. Results The Native-PAGE result showed that there was 1 protein band of the relative molecular mass over 669×103, (232-440)×103 and (140-232)×103,respectively, and 6 bands of the relative molecular mass of (67-140)×103.Besides, 54 proteins were identified with at least 2 distinct peptides in 1 protein band of the relative molecular mass of (232-440)×103, including 4 unnamed protein products, mainly at the isoelectric points of 5.5-8.0 and of the relative molecular mass of (10-40)×103. Based on their functions in the protein database, allthe identified proteins in this study were classified into the following 5 groups: conjugated protein (43%), transport protein (30%), enzyme (6%), signal transducer (4%), and molecular function-unknown protein (17%). At the subcellular localization of the identified proteins, there was mainly a secreted protein (63%), and the remaining proteins were localized in the membrane and cytoplasm. Conclusion Native-PAGE and the Shotgun technique can effectively separate and identify proteins from NRCF, and can identify the components of the protein band of the relative molecular mass of (232-440)×103 and provide basicinformation on the unnamed protein products in NRCF.
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.
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 The article introduces the present status of the application of comparative proteomics in study of tumor marker. Methods This essay review the present status and advances of the application of comparative proteomics in study of tumor marker through refer considerable literatures about proteome, proteomics and tumor marker. Results Follow the study of human genome deepening; the paradox between the finiteness of genes’ number and stability of genes’ structure and the variety of the life phenomena is more conspicuous. Then, the study of proteomics was pushed to the advancing front of life science research. The application of comparative proteomics to tumor research becomes a hot spot nowadays. Conclusion Screening tumor marker via comparative proteomics is an extremely promising research.
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.
Penicillin is a kind ofβ-lactam drug which has been applied in the clinical treatment firstly in the world, and it has still been widely used at present. The synthesis and regulation mechanism of Penicillium chrysogenum, which is used to produce penicillin, has been studied quite maturely, but its proteomics research started relatively late and fewer reports were published. This paper reviews the synthesis and application of penicillin, transformation of Penicillium chrysogenum, and the research progress of its proteomics. On this basis, the study highlights the advantages of proteomics in the research of protein expression.