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Cholangiocarcinoma (CCA) continues to harbor a difficult prognosis and it is difficult to diagnose in its early stages. The molecular mechanisms of CCA oncogenesis and progression are poorly understood. This study aimed to identify candidate biomarkers for CCA. Integrated analysis of microarray data sets was performed to identify differentially expressed genes (DEGs) between CCA and normal tissues. Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were then performed to identify the functions of DEGs. Furthermore, the protein-protein interaction (PPI) network of DEGs was constructed. The expressions of DEGs were validated in human CCA tissues by qRT-PCR. A set of 712 DEGs were identified in CCA compared with normal tissues, including 306 upregulated and 406 downregulated DEGs. It can be shown from the KEGG pathway analysis that some pathways may have important roles in pathology of CCA, including peroxisome proliferator-activated receptor signaling pathway, bile secretion, cell cycle, fat digestion and absorption. PPI network indicated that the significant hub proteins were PKM, SPP1 and TPM1. The abnormally overexpression PKM, SPP1 and TPM1 were closely related to oncogenesis and progression of CCA. PKM, SPP1, TPM1, COL1A1 and COL1A2 may serve as candidate biomarkers for diagnosis and prognosis of CCA.

BACKGROUND Although MTBDRplus is validated for the detection of multidrug-resistant tuberculosis (MDR-TB), its role in the assessment of treatment outcome is less clear. We evaluated the association of MTBDRplus results with treatment outcome in new and previously treated patients in an endemic setting in China and determined factors associated with poor treatment outcomes. METHODS We prospectively enrolled 298 smear-positive pulmonary TB patients who received the World Health Organization recommended initial treatment regimen or retreatment regimen. MTBDRplus was compared with conventional drug susceptibility testing and DNA sequencing for the detection of MDR-TB. Treatment responses were monitored using sputum smear, culture and chest radiography. RESULTS MTBDRplus successfully identified all MDR-TB and had good concordance with sequencing. MDR-TB rates were low among new patients (4/187, 2.1%), but high in previously treated patients (12/28, 42.9%); 65.2% (15/23) of previously treated cases and 17.1% (27/158) of new cases were unsuccessfully treated (P < 0.001). Seven of eight (87.5%) previously treated MDR-TB patients failed the retreatment regimen. In addition to drug resistance, sputum smear positivity at week 8 and cavitation are associated with treatment failure. CONCLUSION Not only did MTBDRplus correctly identify all MDR-TB cases, MTBDRplus results are also associated with treatment outcomes in previously treated patients. The retreatment regimen should no longer be used; treatment should be guided by molecular testing.

Publisher Summary Protein kinase C (PKC) as a collection of Ca2+/phosphatidylserine (PS)/diacylglycerol (DAG)-stimulated kinases has been purified from various sources. The three Ca2+/PS/DAG-stimulated kinases with corresponding Ca2+/PS-dependent phorbol ester-binding activity peaks are designated a PKC I, II, and III. PKC I, II, and III are structurally homologous and behave similarly during purification by ion-exchange, gel-filtration, hydrophobic, and affinity column chromatography. Thus, by using hydroxylapatite column chromatography as a last step of purification, these isozymes are separated and each purified in milligram quantity. For the purification process, fresh rat brains (120 g wet wt) from 80 male Sprague-Dawley rats (200–250 g) are homogenized in 600 ml of ice-cold homogenizing buffer using a Polytron at setting 5 with four 15- sec bursts. The homogenate is centrifuged at 34,000 rpm at 4° for one hr using Beckman 35 rotors. The supernatant fluid is decanted carefully to avoid the turbid fluffy layer and the combined fluffy layer and the pellet are extracted once again with 400 ml of the homogenizing buffer. The combined high-speed supernatant fluid is adjusted to pH 7.5 by adding solid Tris and applied to a DEAE-cellulose (DE-52) column (4.0 × 16 cm) equilibrated with buffer A. The surface of the column is periodically stirred up gently with a glass rod to avoid clotting of the DEAE-cellulose.