Direct assessment of articular cartilage and underlying subchondral bone reveals a progressive gene expression change in human osteoarthritic knees.

OBJECTIVE: To evaluate the interaction of articular cartilage (AC) and subchondral bone (SB) through analysis of osteoarthritis (OA)-related genes of site-matched tissue. DESIGN: We developed a novel method for isolating site-matched overlying AC and underlying SB from three and four regions of interest respectively from the human knee tibial plateau (n = 50). For each site, the severity of cartilage changes of OA were assessed histologically, and the severity of bone abnormalities were assessed by microcomputed tomography. An RNA isolation procedure was optimized that yielded high quality RNA from site-matched AC and SB tibial regions. Quantitative polymerase chain reaction (Q-PCR) analysis was performed to evaluate gene expression of 61 OA-associated genes for correlation with cartilage integrity and bone structure parameters. RESULTS: A total of 27 (44%) genes were coordinately up- or down-regulated in both tissues. The expression levels of 19 genes were statistically significantly correlated with the severity of AC degeneration and changes of SB structure; these included: ADAMTS1, ASPN, BMP6, BMPER, CCL2, CCL8, COL5A1, COL6A3, COL7A1, COL16A1, FRZB, GDF10, MMP3, OGN, OMD, POSTN, PTGES, TNFSF11 and WNT1. CONCLUSIONS: These results provide a strategy for identifying targets whose modification may have the potential to ameliorate pathological alterations and progression of disease in both AC and SB simultaneously. In addition, this is the first study, to our knowledge, to overcome the major difficulties related to isolation of high quality RNA from site-matched joint tissues. We expect this method to facilitate advances in our understanding of the coordinated molecular responses of the whole joint organ.

Prospective study of warfarin dosage requirements based on CYP2C9 and VKORC1 genotypes.

Polymorphisms in CYP2C9 and VKORC1 have been shown to be associated with warfarin dose requirements and could be used to predict warfarin dose. We conducted a prospective study in which warfarin dose was prescribed based on CYP2C9 and VKORC1 polymorphisms in 108 Han-Chinese patients without prior warfarin treatments. Using the genotype-based dosing, 83% of patients reached stable, therapeutic international normalized ratio (INR) within 2 weeks of treatment initiation and none of the patients developed clinical bleeding or thromboembolic event. Ten percent (11) of patients with INR > 4 and no clinical bleeding were detected during this study. At 12 weeks, 69% of the patients' maintenance doses matched the prediction. Dosing algorithms incorporating genetic factors, age, and body surface area were developed, which could explain up to 62% of the total variation (R(2) of 0.62). This study demonstrated that pharmacogenetics-based dosing could improve time to stable, therapeutic INR, reduce adverse events, and achieve high sensitivity.

Study of a three-stage fluidized bed process treating acrylic synthetic-fiber manufacturing wastewater containing high-strength nitrogenous compounds.

Polyacrylonitrile (PAN) is one of the major synthetic fibers commonly used in the mass production of clothing. The chemical synthesis of PAN is carried out by polymerization of the acrylonitrile (AN) monomers with co-monomers such as vinyl acetate, methyl acrylate and cyclohexyl acrylate. Using water quality analysis of the PAN wastewater, high concentration of organic nitrogen was found and the TKN/COD ratios achieved were 0.15-0.26, indicating the complicated biodegradation characteristics for the PAN wastewater. In order to enhance biodegradation of nitrogenous compounds in PAN wastewater, a combined three-stage process of thermophilic anaerobic/anoxic denitrification/aerobic nitrification fluidized bed reactors was employed. The results indicated that the concentration of effluent in the three-stage process of OD and organic nitrogen was 175 mg/L and 13 mg/L, respectively. Furthermore, molecular biotechnology was applied to study the microbial population in the thermophilic anaerobic fluidized bed reactor. From the results of denaturing gradient gel electrophoresis, the diversity of PAN-degrading bacteria would change in different volumetric loading. Furthermore, the bacteria communities in the thermophilic anaerobic fluidized bed reactor were also studied by fluorescence in situ hybridization and confocal laser scanning microscopy. Alpha and delta-Proteobacteria were dominant in the bacteria population, and some high G+C content bacteria and Clostridium could be characterized in this system.