OBJECTIVETo identify potential serum biomarkers for distinguishing between latent tuberculosis infection (LTBI) and active tuberculosis (TB).

METHODSA proteome microarray containing 4,262 antigens was used for screening serum biomarkers of 40 serum samples from patients with LTBI and active TB at the systems level. The interaction network and functional classification of differentially expressed antigens were analyzed using STRING 10.0 and the TB database, respectively. Enzyme-linked immunosorbent assays (ELISA) were used to validate candidate antigens further using 279 samples. The diagnostic performances of candidate antigens were evaluated by receiver operating characteristic curve (ROC) analysis. Both antigen combination and logistic regression analysis were used to improve diagnostic ability.

RESULTSMicroarray results showed that levels of 152 Mycobacterium tuberculosis (Mtb)-antigen- specific IgG were significantly higher in active TB patients than in LTBI patients (P < 0.05), and these differentially expressed antigens showed stronger associations with each other and were involved in various biological processes. Eleven candidate antigens were further validated using ELISA and showed consistent results in microarray analysis. ROC analysis showed that antigens Rv2031c, Rv1408, and Rv2421c had higher areas under the curve (AUCs) of 0.8520, 0.8152, and 0.7970, respectively. In addition, both antigen combination and logistic regression analysis improved the diagnostic ability.

CONCLUSIONSeveral antigens have the potential to serve as serum biomarkers for discrimination between LTBI and active TB.

Adolescent ; Adult ; Aged ; Antibodies, Bacterial ; Antibody Specificity ; Antigens, Bacterial ; Biomarkers ; blood ; Female ; Humans ; Latent Tuberculosis ; blood ; diagnosis ; Logistic Models ; Male ; Middle Aged ; Mycobacterium tuberculosis ; Protein Array Analysis ; methods ; Proteome ; genetics ; Proteomics ; methods ; ROC Curve ; Young Adult

Characterization of the colon cancer immunome and its autoantibody signature from differentially-reactive antigens (DIRAGs) could provide insights into aberrant cellular mechanisms or enriched networks associated with diseases. The purpose of this study was to characterize the antibody profile of plasma samples from 32 colorectal cancer (CRC) patients and 32 controls using proteins isolated from 15,417 human cDNA expression clones on microarrays. 671 unique DIRAGs were identified and 632 were more highly reactive in CRC samples. Bioinformatics analyses reveal that compared to control samples, the immunoproteomic IgG profiling of CRC samples is mainly associated with cell death, survival, and proliferation pathways, especially proteins involved in EIF2 and mTOR signaling. Ribosomal proteins (e.g., RPL7, RPL22, and RPL27A) and CRC-related genes such as APC, AXIN1, E2F4, MSH2, PMS2, and TP53 were highly enriched. In addition, differential pathways were observed between the CRC and control samples. Furthermore, 103 DIRAGs were reported in the SEREX antigen database, demonstrating our ability to identify known and new reactive antigens. We also found an overlap of 7 antigens with 48 "CRC genes." These data indicate that immunomics profiling on protein microarrays is able to reveal the complexity of immune responses in cancerous diseases and faithfully reflects the underlying pathology.
Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; genetics ; immunology ; metabolism ; Case-Control Studies ; Colonic Neoplasms ; immunology ; metabolism ; Computational Biology ; methods ; Computer Simulation ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Immunoglobulin G ; immunology ; Male ; Middle Aged ; Protein Array Analysis ; methods

OBJECTIVETo investigate the protein markers in the urine of chronic renal failure (CRF) patients of Chinese medicine damp syndrome (CMDS) based on surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) technique.

METHODSThe urine was sampled from 90 CRF patients of CMDS and 60 CRF patients of non-CMDS. Then the proteomics of the urine was studied by H4 gene chip. The chips were scanned and analyzed using PBS II (a protein chip reader).

RESULTS(1) Totally 25 differential protein peaks were identified in the e/m range of 1 000-20 000 of the protein atlas of the two groups (P < 0.01). (2) The urine protein predictive model of CFR patients of CMDS was established after bioinformatic analysis. Totally 4 biomarkers consisting of M/Z 8 654.96, M/Z 2 081.65, M/Z 18 667.3, and M/Z 2 242.14 were obtained, which could better classify the samples of CMDS and those of non-CMDS. Its accuracy rate was 84.7%, the sensitivity was 92.2%, and the specificity was 73.3%. (3) Between the CMDS group and the non-CMDS group, 7 kinds of proteins in the urine were possibly identified by SwissProt Database.

CONCLUSIONSThis study had primarily screened the protein markers in the urine of CRF patients of CMDS, and established the predictive model. The protein markers in the urine were identified by database, thus providing certain experimental evidence for clinical typing of CRF patients of CMDS.

Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Kidney Failure, Chronic ; diagnosis ; urine ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Protein Array Analysis ; methods ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

OBJECTIVETo establish serum protein fingerprint model for early diagnosis of pancreatic cancer with surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS) and bioinformatics techniques.

METHODSA total of 73 samples were analyzed in this study, including 31 cases of pancreatic cancers, 22 cases of pancreatitis and 20 healthy individuals. Samples were first analyzed by SELDI-TOF-MS and two patterns of differentiation model were constructed with support vector machine arithmetic method.

RESULTSThe pattern 1 model differentiating pancreatic cancer patients from healthy individuals had a specificity and a sensitivity of both 100.0%. The pattern 2 model differentiating pancreatic cancer from pancreatitis had a specificity of 95.5% and a sensitivity of 93.5%.

CONCLUSIONSELDI-TOF-MS technique combined with bioinformatics can facilitate to identify biomarkers for pancreatic cancer.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; blood ; diagnosis ; Protein Array Analysis ; methods ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Support Vector Machine

OBJECTIVETo establish and evaluate a protein microarray method for combined measurement of serum ferritin (SF) and soluble transferrin receptor (sTfR).

METHODSMicroarrayer was used to print both anti-SF antibodies I and anti-sTfR antibodies I on each protein microarray. Anti-SF antibodies II and anti-sTfR antibodies II were used as detection antibodies and goat antibodies coupled to Cy3 were used as antibodies III. The detection conditions of the quantitative analysis method for simultaneous measurement of SF and sTfR with protein microarray were optimized and evaluated. The protein microarray was compared with commercially available traditional tests with 26 serum samples.

RESULTSBy comparison experiment, mouse monoclonal antibodies were chosen as the probes and contact printing was chosen as the printing method. The concentrations of SF and sTfR probes were 0.5 mg/mL and 0.5 mg/mL respectively, while those of SF and sTfR detection antibodies were 5 μg/mL and 0.36 μg/mL respectively. Intra- and inter-assay variability was between 3.26% and 18.38% for all tests. The regression coefficients comparing protein microarray with traditional test assays were better than 0.81 for SF and sTfR.

CONCLUSIONThe present study has established a protein microarray method for combined measurement of SF and sTfR.

Animals ; Antibodies, Monoclonal ; analysis ; Ferritins ; blood ; Mice ; Protein Array Analysis ; methods ; Rabbits ; Receptors, Transferrin ; blood

BACKGROUNDThe expression of genes encoding a number of pathogenetic pathways involved in colorectal cancer could potentially act as prognostic markers. Large prospective studies are required to establish their relevance to disease prognosis.

METHODSWe investigated the relevance of 19 markers in 790 patients enrolled in a large randomised trial of 5-fluorouracil using immunohistochemistry and chromogenic in situ hybridisation. The relationship between overall 10-year survival and marker status was assessed.

RESULTSMinichromosome maintenance complex component 2 (MCM2) and cyclin A were significantly associated with overall survival. Elevated MCM2 expression was associated with a better prognosis (HR = 0.63, 95%CI: 0.46 - 0.86). Cyclin A expression above the median predicted an improved patient prognosis (HR = 0.71, 95%CI: 0.53 - 0.95). For mismatch repair deficiency and transforming growth factor β receptor type II (TGFBRII) overexpression there was a borderline association with a poorer prognosis (HR = 0.69, 95%CI: 0.46 - 1.04 and HR = 2.11, 95%CI: 1.02 - 4.40, respectively). No apparent associations were found for other markers.

CONCLUSIONThis study identified cell proliferation and cyclin A expression as prognostic indicators of patient outcome in colorectal cancer.

Aged ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; Cyclin A ; metabolism ; DNA Mismatch Repair ; genetics ; physiology ; Female ; Humans ; In Situ Hybridization ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Prognosis ; Prospective Studies ; Protein-Serine-Threonine Kinases ; metabolism ; Receptors, Transforming Growth Factor beta ; metabolism ; Tissue Array Analysis ; methods

OBJECTIVETo study the value of multiple Helicobacter pylori (H.pylori) antibody detection by protein array in the diagnosis of H.pylori infection in children.

METHODSBiopsy specimens obtained by gastroscopy from 120 children with digestive system symptoms were detected by rapid urease test (RUT) and modified Giemsa staining. Positivity in both RUT and Giemsa staining was the "gold criterion" of H.pylori infection. Serum samples of these patients were obtained and the antibodies against cytotoxin associated gene A protein (CagA), vacuolating toxin A (VacA), urease, heat shock protein 60 (Hsp60) and RdxA (nitroreductase) were detected by protein array technique.

RESULTSH.pylori infection was identified according to the "gold criterion" in 60 children. Compared with the "gold criterion", the goodness of fit and the coefficient of contingency in the diagnosis of H.pylori infection of the following four groups antibody detection were all statistically significant (P<0.001): anti-Ure antibody alone, anti-Ure antibody combined with anti-CagA antibody, anti-Ure antibody combined with anti-VacA antibody and anti-Ure antibody combined with anti-CagA and anti-VacA antibody. The sensitivity, specificity and accuracy of the detection of anti-Ure antibody combined with anti-CagA antibody for the diagnosis of H.pylori infection were 81.7%, 91.7% and 86.7%, respectively. The antibody detection showed a high positive predictive value (90.7%) and a high negative predictive value (83.3%).

CONCLUSIONSThe antibody detection by protein array, especially the detection of anti-Ure antibody combined with anti-CagA antibody, is valuable in the diagnosis of H.pylori infection.

Adolescent ; Antibodies, Bacterial ; blood ; Child ; Child, Preschool ; Female ; Helicobacter Infections ; diagnosis ; Helicobacter pylori ; immunology ; Humans ; Male ; Protein Array Analysis ; methods ; Sensitivity and Specificity

Semiconductor quantum dots (QDs) are a new kind of biological fluorescence material, which has many advantages, such as broad excitation spectra, tunable emission spectra and good photostability. In the field of biomedical detection, the problems encountered in the traditional organic dye-based biomedical detections, such as short fluorescence lifetime and failure to simultaneous excitation of multiple colors, can be solved by using QDs. Water-soluble QDs combined with specific bio-molecules can label targeting bio-compound, which is useful in bio-molecule detection, cell labeling, tissue imaging, and can be used in fluorescence resonance energy transfer (FRET) technology. Combining QDs and protein chip technology to develop a new technology to detect multiple kinds of tumor markers will be one of the promising clinical applications of QDs with greater sensitivity, specificity, rapidity and convenience.
Animals ; Biomarkers, Tumor ; analysis ; Fluorescence Resonance Energy Transfer ; methods ; Humans ; Protein Array Analysis ; Quantum Dots ; Sensitivity and Specificity

Medical community and pharmaceutical companies are currently facing a dire need for discovery and identification of new druggable targets. However, the discovery of small-molecule target is an important and arduous task for the biological and medical scientists. To overcome the bottlenecks for target validation, many new approaches are being developed, such as chemical proteomics. As a part of proteomics approaches, chemical proteomics employs small-molecule compounds that can specifically interact with the target protein to interfere with and detect proteomics. Therefore, new target identification, drug discovery and research on multi-target-directed drugs will all be benefited from the further advances in chemical proteomics approaches. Chemical proteomics has the potential to greatly enhance the efficiency of the drug discovery process.
Animals ; Drug Delivery Systems ; methods ; Drug Design ; Drug Discovery ; Drugs, Chinese Herbal ; chemistry ; Humans ; Protein Array Analysis ; Proteomics ; methods ; Small Molecule Libraries ; chemistry

OBJECTIVETo investigate the effects of a microfluidic sperm sorter on the routine parameters and DNA integrity of human sperm.

METHODSWe divided 40 semen samples into two aliquots and performed sperm sorting using a self-made polydimethylsiloxane microfluidic sperm sorter and the swim-up method, respectively. Then we evaluated and compared the effects of these two methods on the sperm routine parameters and DNA integrity by computer-assisted sperm analysis and sperm chromatin dispersion test.

RESULTSAfter processing, sperm motility, normal morphology and tail hypoosmotic swelling rate were significantly improved, while sperm DNA damage remarkably decreased (P < 0.01). The microfluidic sperm sorter achieved a significantly lower rate of sperm DNA damage than the swim-up method ([ 8.4 +/- 5.8 ]% vs [16.4 +/- 9.2] %, P < 0.01), but no statistically significant differences were found in all other parameters between the two methods.

CONCLUSIONHigh-quality sperm with less DNA integrity damage could be obtained in sperm sorting with the microfluidic sperm sorter.

Adult ; Cell Separation ; instrumentation ; methods ; DNA ; DNA Damage ; Humans ; Infertility, Male ; genetics ; physiopathology ; Male ; Microfluidics ; Middle Aged ; Protein Array Analysis ; Semen Analysis ; instrumentation ; methods ; Sperm Motility ; Spermatozoa