Application of urinary proteomics in early diagnosis of bladder urothelial carcinoma / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the difference in urinary proteome between patients with bladder urothelial carcinoma (BUC) and healthy volunteers and to provide a basis for the early diagnosis of BUC.</p><p><b>METHODS</b>The urine samples from BUC patients and healthy volunteers (controls) were treated by 25% ethanol precipitation and two-dimensional gel electrophoresis (2-DE), and the obtained urinary proteins were subjected to Coomassie brilliant blue staining and analysis by PDQuest 8.0 (2-DE image analysis software); the differentially expressed proteins were sequenced by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry and identified using the Swiss-Prot database; the differential expression of these proteins was verified by western blot.</p><p><b>RESULTS</b>High-resolution and high-reproducibility 2-DE images were obtained from the urine samples of BUC patients and controls, with 789 ± 18 and 762 ± 14 protein spots, respectively. Compared with the control group, the BUC grouP had significantly decreased expression of 6 protein spots and significantly increased expression of 11 protein spots. The mass spectrometry revealed five proteins with increased expression in the BUC group, including fibrinogen, lactate dehydrogenase B, apolipoprotein A1, clusterin, and haptoglobin, and the results were confirmed by western blot.</p><p><b>CONCLUSION</b>There is significant difference in urinary proteome between BUC patients and healthy volunteers; the identification of differentially expressed proteins in urine lays the foundation for identifying potential molecular markers in early diagnosis of BUC.</p>

Significance of apolipoprotein A1 as biomarker for early diagnosis and classification of bladder urothelial carcinoma / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the significance of apolipoprotein (Apo)-A1 in urine as a biomarker for early diagnosis and classification of bladder urothelial carcinoma (BUC).</p><p><b>METHODS</b>Urine samples were divided into four groups: normal control group, benign bladder disease group, low-grade malignant BUC group, and high-grade malignant BUC group. Apo-A1, which showed significantly different expression among the four groups, was selected according to the two-dimensional electrophoresis (2-DE) images of the four groups, and enzyme-linked immunosorbent assay (ELISA) was used to quantify Apo-A1 in the four groups. A receiver operating characteristic (ROC) curve was generated, and the optimal operating points on the ROC curve were found to determine the critical concentrations of Apo-A1 for early diagnosis of BUC and differentiation of low-grade and high-grade malignant BUC. The results were verified clinically, and the specificity and sensitivity were calculated.</p><p><b>RESULTS</b>The 2-DE images showed that that the level of Apo-A1 increased from the normal control grouP to high-grade malignant BUC group. The ELISA showed that there was no significant difference in Apo-A1 level between the normal control grouP and benign bladder disease group, but the Apo-A1 level was significantly higher in the BUC groups than in the normal control grouP and benign bladder disease grouP (P < 0.01); the high-grade BUC grouP had a significantly higher Apo-A1 level than the low-grade BUC grouP (P < 0.01). The BUC patients and those without BUC could be differentiated with an Apo-A1 concentration of 18.22 ng/ml, while the low-grade and high-grade malignant BUC could be differentiated with an Apo-A1 concentration of 29.86 ng/ml. When used as a biomarker, Apo-A1 had a sensitivity of 91.6% (98/107) and a specificity of 85.7% (42/49) for diagnosis of BUC and had a sensitivity of 83.7% (41/49) and a specificity of 89.7% (52/58) for BUC classification.</p><p><b>CONCLUSION</b>Apo-A1 may be a biomarker for early diagnosis and classification of BUC and shows promise for clinical application.</p>

Primary study on changes of serum proteomics in rabbit superior mesenteric artery occlusion shock / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explor the changes of serum proteomics in rabbits superior mesenteric artery occlusion (SMAO) shock as well as its possible effect in SMAO shock.</p><p><b>METHODS</b>SMAO shock model in rabbits were induced by occlusion of the superior mesenteric artery, serum samples were obtained from rabbits before and after SMAO shock, proteins in samples were separated by two-dimensional electrophoresis(2-DE), spots in the 2-DE map were detected and evaluated by PDQuest software 8.0. The spots with different expression level were subjected to matrix assisted laser desorption/ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF-TOF-MS) for identification, the protein database was searched to further characterized the differential proteins.</p><p><b>RESULTS</b>19 differential protein spots were screened out in the 2-DE maps, 11 proteins were up-regulated and 8 proteins were down-regulated in SMAO shock rabbits' s serum. 4 of the 19 differential protein spots were selected for MALDI-TOF-TOF-MS study, and 2 of the 4 differential protein spots were identified satisfactoryly as paraoxonase and haptoglobin, which content were increased in rabbits' s serum after SMAO shock.</p><p><b>CONCLUSION</b>Serum proteomics of rabbit change remarkablely before and after SMAO shock, paraoxonase and haptoglobin may be associated with the compensation after SMAO shock.</p>

Relationship and clinical significance of KiSS-1, nuclear factor kappa B (NF-kappaB), p50, and matrix metalloproteinase 9 expression in breast cancer / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the expression of KiSS-1, nuclear factor kappa B (NF-kappaB) p50 and matrix metalloproteinase 9 (MMP-9) in breast cancer tissue and the relationship with clinicpathological factors.</p><p><b>METHODS</b>Immunohistochemical staining for KiSS-1, NF-KappaBp50, and MMP-9 protein was performed in 152 cases of human breast tissue [92 cases of BC, 30 cases of epithelial hyperplasia, and 30 cases of peritumoral breast tissue (PMT)] and 54 cases of axillary lymph node metastases. In-situ hybridization for KiSS-1 mRNA was done in 50 cases of breast cancer, and 20 cases of PMT.</p><p><b>RESULTS</b>(1) The expression of KiSS-1 gene was significantly higher in well-differentiated breast cancer than in PMT, and this expression progressively decreased with decreasing degree of tumor differentiation, increasing pathological grade, TNM stage and the presence of lymph node metastases. The expression of KiSS-1 gene in lymph node metastasis was markedly lower than the corresponding primary tumor. There was correlation between the expression of KiSS-1 mRNA and KiSS-1 protein in breast cancer group. (2) The expression of NF-kappaKBp50 and MMP-9 increased progressively with decreasing degree of tumor differentiation, increasing TNM stage, large tumor size ( >2 cm) and the presence of lymph node metastases.</p><p><b>CONCLUSIONS</b>The expression of KiSS-1 protein showed negative correlation with that of NF-kappaBp50 and MMP-9 respectively. MMP-9 protein expression was positively correlated with NF-kappap50 protein expression. These suggest that the genes of KiSS-1, NF-kappaBp50 and MMP-9 could be involved in the progression and metastasis of breast cancer.</p>