RESUMO
Temporomandibular joint osteoarthritis (TMJ OA) is a common and heterogeneous disease that causes painful and progressive joint degeneration, which restricts daily activities, including talking and chewing. Long noncoding RNAs (lncRNAs) are an important class of genes involved in various physiological and pathological functions, including osteoarthritis (OA).The present study aimed to identify the lncRNAs that are important in TMJ OA and their potential functions. Here, we found that HOTAIR was significantly upregulated in the synovial fluid of TMJ OA patients compared with that of normal controls. Increased HOTAIR was similarly observed in the synovial fluid of TMJ OA rabbits as compared to control rabbits. Furthermore, in interleukin-1ß (IL-1ß)-induced TMJ OA in vitro model (primary rabbit condylar chondrocytes), the expressions of matrix metalloproteinase (MMP)-1, MMP3, MMP9 and HOTAIR were all dramatically increased. Most importantly, knockdown of HOTAIR in IL-1ß-induced TMJ OA in vitro model could not only reverse the IL-1ß-stimulated expressions of MMP1, MMP3 and MMP9, but also significantly decrease the apoptosis rate induced by IL-1ß in primary rabbit condylar chondrocytes. Our data provides new insight into the mechanisms of chondrocytes destruction in TMJ OA.
Assuntos
Condrócitos/metabolismo , Interleucina-1beta/farmacologia , Metaloproteinases da Matriz/genética , Osteoartrite/genética , RNA Longo não Codificante/metabolismo , Transtornos da Articulação Temporomandibular/genética , Adulto , Animais , Apoptose , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/enzimologia , Condrócitos/patologia , Regulação Enzimológica da Expressão Gênica , Humanos , Metaloproteinases da Matriz/biossíntese , Pessoa de Meia-Idade , Osteoartrite/enzimologia , Osteoartrite/metabolismo , Osteoartrite/patologia , Coelhos , Líquido Sinovial/metabolismo , Transtornos da Articulação Temporomandibular/enzimologia , Transtornos da Articulação Temporomandibular/metabolismo , Transtornos da Articulação Temporomandibular/patologia , Regulação para CimaRESUMO
BACKGROUND: Rapid advances in multislice computed tomography (MSCT) technology facilitate accurate clinical imaging. The newly developed 64-slice CT increases temporal and spatial resolution efficiently. PURPOSE: The purpose of this study is to evaluate the application of 64 slice spiral computed tomography (CT) on the imaging of the normal optics canal. METHODS AND MATERIALS: 100 healthy adults were investigated using 64 slice spiral CT. The optics canal was scanned, reconstructed and examined. RESULTS: Among the four walls of the optic canal, the medial wall is the longest one. The upper wall and outer wall are inferior to the medial wall while the inferior wall is the shortest one. All the data accomplished by the 64 slice CT was consistent with the results of previous reports using other methods. CONCLUSION: The results suggested that the 64 slice spiral CT could be a valuable and accurate method for measuring the length of optics canal walls.
RESUMO
A gene encoding the trehalose phosphorylase (TreP), which reversibly catalyzes trehalose degradation and synthesis from alpha-glucose-1-phosphate (alpha-Glc-1-P) and glucose, was cloned from Thermoanaerobacter tengcongensis and successfully expressed in Escherichia coli. The overexpressed TreP, with a molecular mass of approximately 90 kDa, was determined by SDS-PAGE. It catalyzes trehalose synthesis and degradation optimally at 70 degrees C (for 30 min), with the optimum pHs at 6.0 and 7.0, respectively. It is highly thermostable, with a 77% residual activity after incubation at 50 degrees C for 7 h. Under the optimum reaction conditions, 50 microg crude enzyme of the TreP is able to catalyze the synthesis of trehalose up to 11.6 mmol/L from 25 mmol/L alpha-Glc-1-P and 125 mmol/L glucose within 30 min, while only 1.5 mmol/L out of 250 mmol/L trehalose is degraded within the same time period. Dot blotting revealed that the treP gene in T. tengcongensis was upregulated in response to salt stress but downregulated when trehalose was supplied. Both results indicate that the dominant function of the T. tengcongensis TreP is catalyzing trehalose synthesis but not degradation. Thus it might provide a novel route for industrial production of trehalose.
Assuntos
Glucosiltransferases/genética , Thermoanaerobacter/genética , Cromatografia Líquida de Alta Pressão , Eletroforese em Gel de Poliacrilamida , Estabilidade Enzimática , Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Glucose/metabolismo , Glucofosfatos/metabolismo , Glucosiltransferases/metabolismo , Concentração de Íons de Hidrogênio , Cinética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidade por Substrato , Temperatura , Thermoanaerobacter/enzimologia , Trealose/metabolismoRESUMO
The lipA gene encoding a thermostable esterase was cloned from Thermoanaerobacter tengcongensis and over-expressed in Escherichia coli. The recombinant esterase, with a molecular mass of approx. 43 kDa determined by SDS-PAGE, was purified to homogeneity through Sephadex G-100 gel filtration. The purified enzyme actively hydrolyzed tributyrin but not olive oil. Maximum activity was observed on p-nitrophenyl (NP)-propionate (C3) and p-NP-butyrate (C4), with little activity towards p-NP-palmitate (C16). The esterase was optimally active at 70 degrees C (over 15 min) and at pH 9. It is highly thermostable, with a residual activity greater than 80% after incubation at 50 degrees C for more than 10 h. The activity was not inhibited by 5 mM EDTA and PMSF, indicating the esterase is not a metalloenzyme and may contain a specific structure around the catalytic serine residue. In addition, it was stable for 1 h at 37 degrees C in 1% CHAPS and Triton X-100 but not stable in 1% Tween 20 or SDS.
