ABSTRACT
Objective To analyze the differentially expressed proteins in bone marrow supernatants of multiple myeloma patients by using 2-DE and MALDI-TOF-MS,and search for the special protein markers for studying the mechanism of the development and diagnosis or differential diagnosis of multiple myeloma.Methods The bone marrow supernatant samples of fourteen multiple myeloma patients,five other hematologic malignancies and five normal controls were collected.After removing albumin and IgG,the proteins in the supernatants were separated by 2-DE.Three groups images were analyzed and compared by Imagemnster 2D platinum 5.0 analysis software.Differentially expressed proteins were selected if the protein spots intensity showed more than 3 fold increase or decrease among different groups.The identities of the differentially expressed proteins with good repeatibility were determined by PMF based on by MALDI-TOF-MS or MALDI-TOF-MS/MS and NCBInr database search.Results 2-DE maps of bone marrow supernatants of the three groups could be analyzed and compared by image analysis of software.Forty-seven and fifty-eight differentially expressed protein spots were detected in multiple myeloma samples compared with normal controls and other hematologic malignancies samples,respectively.Forty-one reproducible spots were analyzed and identified by mass spectrum.Compared with other hematologic malignancies and normal controls,five up-expressed proteins and three down-expressed proteins were identified in multiple myeloma samples.They includes immunoglobulin J chain κ and λ light chain,provirus ancestral Gag polyprotein,mature oxy-cope catalytic antibody with hapten for up-expressed proteins,and hemoglobin,haptoglobin Hp2,zinc-alPha-2-glycoprotein for down-expressed proteins.These differentially expressed proteins reflect the features of muhiple myeloma,and relate to the development,progression and therapy of multiple myeloma.Conclusions Eight differentially expressed proteins in bone marrow supernatants of multiple myeloma are identified by using 2-DE and MALDI-TOF-MS.These differentially expressed proteins could be useful in studying the mechanism of the development and progression of multiple myeloma,and in developing diagnosis and differential diagnosis of multiple myeloma.
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Objective To assess the levels of glucose-6-phosphate isomerase(GPI) mRNA in peripheral blood monocytes and serum GPI levels in patients with rheumatoid arthritis, and analyze the association of serum GPI with MCV antibody, CCP antibody and RF of RA. Methods Fluorogenic quantitative real-time polymerase chain reaction (FQ-RT-PCR) was used to examine mRNA expression on peripheral blood monocytes in 60 RA patients (28 case in active stage,32 cases in stable stage) ,30 patients with other rheumatic diseases, and 30 healthy controls. ELISA was used to detect the levels of serum GPI, anti-MCV antibodies, anti-CCP antibodies and RF in each group. Results The levels of GPI mRNA in RA group [△Ct=4.21 (3.04-7.23)] were significantly higher than those in patients with other rheumatic diseases [△Ct=8.42 (5.16-9.98),P<0.01] and healthy controls [△Ct=8.66 (4.90-10.01), P<0.01]. There were statistically significant differences of GPI mRNA levels between active RA [△Ct=3.78 (1.28-6.09)] and inactive RA[△Ct =5.88(3.23-8.94),H=11.760,P<0.01)]. The RA group serum GPI levels [3.02 (2.02-8.39) mg/L] were higher than those of other rheumatic diseases [0.20 (0.11-0.32) mg/L] and healthy controls [0.18(0.08-0.30) mg/L]. There were significant differences of serum GPI levels between active RA group [4.84(2.81-10.38) mg/L] and inactive RA group[2.12 (1.26-4.34) mg/L] (H=9.830, P<0.01). The sensitivities of GPI, anti-MCV and anti-CCP were 68% (41/60) ,57% (34/60),58% (35/60), respectively and specificities were 95% (57/60), 92% (55/60) and 93% (56/60), respectively. Conclusions The high expression of GPI mRNA in RA patients shows that it may play a pathological role in the development of RA, and it may be correlated with the activity of RA. It may be a valuable diagnostic parameter for RA, because of its high sensitivity and specificity.
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Objective To detect the expression of GITR mRNA in peripheral blood mononuclear cells (PBMCs) of patients with primary biliary cirrhosis (PBC) and its relationship with the disease activity of PBC.Methods Real time quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) was used to examine GITR mRNA expression on peripheral blood monocytes of 44 active PBC, 34 stable PBC, 30.hepa-toma cases and 30 healthy controls. Results The mean GITR mRNA expression of active PBC (△Ct=10.5±