Increased expression of serum gelsolin in patients with osteosarcoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Identification of potential serum biomarkers of osteosarcoma to aid in its early diagnosis and in the discovery of possible therapeutic targets is an area of increasing interest.</p><p><b>METHODS</b>Two-dimensional difference-in-gel electrophoresis was used to assess multiple serum samples in patients with osteosarcoma. In addition, differential expression of protein biomarkers was characterized in osteosarcoma serum by using matrix-assisted desorption/ionization time-of-flight mass spectrometry coupled with database interrogation. Serum samples from four individuals with osteosarcoma and four age- and sex-matched healthy control subjects were compared.</p><p><b>RESULTS</b>Fifty-eight significant protein spot features in the osteosarcoma sera were found. These spot features were excised, digested with trypsin, and analyzed with mass spectrometry. Gelsolin was down-regulated only in osteosarcoma. Furthermore, Western blotting and enzyme linked immunosorbent assay (ELISA) confirmed decreased levels of gelsolin in the osteosarcoma serum samples.</p><p><b>CONCLUSIONS</b>These results indicated that gelsolin might have great potential as a biomarker of osteosarcoma and as a potential target for gene therapy.</p>

Proteomic analysis between keloid and normal skin / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate and search correlative proteins of keloid by comparing the results of differential proteomic analysis between keloid and normal skin.</p><p><b>METHODS</b>From January 2010 to June 2010 two-dimensional gel electrophoresis was used to define patterns of protein expression in keloid skin from 8 patients and matched normal skin from 3 patients. Differential expression protein spots were showed and analyzed by matrix-assisted laser desorption ionization-time of flying/time of flying (MALDI-TOF/TOF) mass spectrometry.</p><p><b>RESULTS</b>This study succeeded to provide a two-dimensional protein profiling comparison between normal skin and keloid. Gel-analysis software identified an average of 2978 spots in keloid while 3053 spots in normal skin and statistical filtering yielded 40 spots of a 4-fold change, 32 of which were identified by using mass spectrometry, 20 were up-regulated and 12 were down-regulated. Functional analysis revealed that these proteins could be fractionated to carrier proteins (3 proteins), signal transduction proteins (4 proteins), proliferation and apoptosis related proteins (2 proteins), cytoskeleton proteins (6 proteins), extracellular matrix proteins (8 proteins), immunity related proteins (3 proteins), tumor related proteins (2 proteins), and function unknown protein (4 proteins).</p><p><b>CONCLUSIONS</b>Proteomic analysis can identify the proteins with variance of keloid versus normal skin. The further research to these differential proteins may help reveal the pathogenesis of keloid and provide new treatments for keloid.</p>

Comparison of two techniques for expression and purification of glycogen synthase kinase 3β / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a method for the expression of glycogen synthase kinase 3β with high purity and biological activity.</p><p><b>METHODS</b>E.coli expression system and baculovirus-insect cell expression system were used to produce the kinase, followed by purification using His-tag and GST-tag and determination of its purity and activity by SDS-PAGE and kinase reaction, respectively.</p><p><b>RESULTS</b>Glycogen synthase kinase 3β produced from E.coli represented 54% of the total bacterial protein, as compared with 96% of the total protein from the insect cell system .Glycogen synthase kinase 3β produced from insect cell exhibited an one-fold higher biological activity than the protein obtained from E.coli.</p><p><b>CONCLUSIONS</b>Compared with the protein from E.coli system, glycogen synthase kinase 3β from the insect cell expression system is endowed with a higher purity and bioactivity.</p>

Expression of CD45 in the serum of patients with Crohn's disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To search for differentially expressed proteins in the serum of patients with Crohn's disease.</p><p><b>METHODS</b>Serum protein samples obtained from 4 patients with Crohn's disease and 4 normal adults were cross-labeled with different CyDyes and underwent two-dimensional differential in-gel electrophoresis (2-D DIGE) and imaging analysis. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to identify the differentially expressed proteins.</p><p><b>RESULTS</b>2-D DIGE revealed that the protein on spot 973 was overexpressed by 2.55 folds in the serum of patients with Crohn's disease compared with that in normal adults (P<0.05). The protein was identified as CD45 using mass spectrometry.</p><p><b>CONCLUSION</b>CD45 overexpression in the serum of patients with Crohn's disease may play a role in the disequilibrium of the immune system.</p>

Difference of gene expression between the central and the peripheral epithelia of the bovine lens / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Equatorial lens epithelial cells proliferate and differentiate into fiber cells throughout life, while central lens epithelial cells proliferate little and do not form fiber cells. This study aimed to investigate the differences in gene expression between the central and the peripheral epithelial cells of the bovine lens.</p><p><b>METHODS</b>Lens epithelia were dissected into central (<or= 11.5 mm diameter, cLEC) and peripheral regions (pLEC). The differences in gene expression and protein accumulation between these two regions were assayed by microarray analysis and two-dimensional electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Differently expressed proteins were validated by immunoanalyses.</p><p><b>RESULTS</b>By microarray analysis, 67 transcripts were at least two-fold lower and 269 at least two-fold higher in pLEC compared with that in cLEC. Thirty-four protein spots, including 20 in cLEC and 14 in pLEC, were identified by two dimensional electrophoresis and mass spectrometry. Of these 34 protein products, 28 were represented by probe sets on the microarray. Nine transcripts changed in the same direction and four transcripts in the opposite direction to their protein products. Immunoanalyses revealed that three (mitogen-activated protein kinase 1 (MAPK1), nidogen (NID), small nuclear ribonucleoprotein N (SNRPN)) out of four transcripts with opposite change between 2-DE and microarray assay showed the same changes as the results of 2-DE gel analyses. The genes differently expressed between cLEC and pLEC mainly include those related to the MAPK, transforming growth factor beta (TGFbeta) signaling and glycolysis pathways.</p><p><b>CONCLUSION</b>The results suggested that there were distinctly different genome activities, including a specific group of pathways, between central and peripheral lens epithelial cells.</p>

Detection of serum biomakers of osteosarcoma by proteomic profiling / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect differentially expressed proteins in serum of patient with osteosarcoma.</p><p><b>METHODS</b>8 serum protein samples were recruited (4 cases of osteosarcoma and 4 cases of normal adults), cross-labeled with variant CyDye, followed by two-dimensional differential in-gel electrophoresis (2-D DIGE), image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).</p><p><b>RESULTS</b>24 protein spot-features were significantly increased, and 34 were significantly decreased in the serum from patients with osteosarcoma relative to the controls. The mass spectrometry analysis revealed 18 unique proteins that were increased, and 25 unique proteins decreased in the serum of patients with osteosarcoma. Gelsolin was down-regulated in osteosarcoma, and Western blotting also confirmed a decreased level of gelsolin in the serum of patients with osteosarcoma.</p><p><b>CONCLUSION</b>Our results indicate that gelsolin may have great potential as a biomarker of osteosarcoma and as a potential target for therapy. These preliminary data suggest that incorporation of more samples and new datasets will permit the identification of serum biomarkers for osteosarcoma.</p>

Serum proteomic variation study in patients with Crohn disease / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To search differentially expressed proteins in serum of patients with Crohn disease.</p><p><b>METHODS</b>Serum protein samples from 4 patients with Crohn disease and 8 healthy adults were recruited cross-labeled with variant CyDye, and then followed by two-dimensional differential in-gel electrophoresis (2-D DIGE), image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).</p><p><b>RESULTS</b>The 2-D electrophoresis results were compared between the Crohn disease patients and the healthy adults. The spot 1058 expression in serum of Crohn disease patients increased by 1.68 folds as compared with healthy adults (P<0.05). The protein was identified as haptoglobin by mass spectrometry.</p><p><b>CONCLUSION</b>Up-regulating expression of haptoglobin in serum of Crohn disease patients may play a role in disequilibrium of immunity system.</p>

Role of phospho-calcium/ calmodulin-dependent protein kinase II in the induction and maintenance of long-term potentiation of C-fiber-evoked field potentials in spinal dorsal horn of the rat / 生理学报

Our previous studies have shown that long-term potentiation (LTP) of C-fiber-evoked field potentials in the spinal dorsal horn is NMDA receptor dependent. It is known that elevation of Ca(2+) in the postsynaptic neurons through NMDA receptor channels during high-frequency stimulation of the afferent fibers is crucial for LTP induction, but how this leads to a prolonged potentiation of synaptic transmission in the spinal dorsal horn is not clear. In the hippocampus, a rise of Ca(2+) activates calcium/calmodulin-dependent protein kinase II (CaMK II) through autophosphorylation. Once this occurs, the kinase remains active, even when Ca(2+) concentration returns to baseline level. Phosphorylated CaMK II potentiates synaptic transmission by enhancement of AMPA receptor channel function via phosphorylation of GluR1 subunit of the receptor and the addition of AMPA receptors to synapses. Up to now, the role of CaMK II in the induction and maintenance of LTP of the C-fiber-evoked field potentials in spinal dorsal horn has not been evaluated. In the present study, we examined the expression of CaMK II and phospho-CaMK II in the lumbar segments (L4-L6) of the rat spinal dorsal horn at 30 min and 3 h after the establishment of LTP induced by tetanic electrical stimulation of the sciatic nerve (40 V, 0.5 ms pulses at 100 Hz for 1 s repeated four times at 10 s intervals) by using Western blot and electrophysiological techniques. To determine the role of the phospho-CaMK II in the induction and maintenance of the spinal LTP, a selective CaMK II inhibitor KN-93 (100 micromol/L) was applied directly onto the spinal cord at the recording segments before and after LTP induction. We found that (1) the protein level of phospho-CaMKII increased at both 30 min and 3 h after LTP induction, while the total protein level of CaMK II increased at 3 h but not at 30 min after LTP induction. (2) Spinal application of KN-93 at 30 min prior to the tetanus blocked both LTP induction and the increase in phospho-CaMK II. (3) 30 min after LTP induction, spinal application of KN-93 depressed LTP and the level of phospho-CaMK II (n=3). (4) Spinal application of KN-93 at 3 h after LTP, however, affected neither the amplitude of the spinal LTP nor the level of phospho-CaMK II in the spinal dorsal horn. These results suggest that activation of CaMK II is probably crucial for the induction and the early-phase maintenance of LTP of C-fiber-evoked field potentials in the spinal dorsal horn.