Cangxi Tongbi Capsules promote chondrocyte autophagy by regulating circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit development of knee osteoarthritis / 中国中药杂志

To investigate the mechanism by which Cangxi Tongbi Capsules promote chondrocyte autophagy to inhibit knee osteoarthritis(KOA) progression by regulating the circRNA＿0008365/miR-1271/p38 mitogen-activated protein kinase(MAPK) pathway. The cell and animal models of KOA were established and intervened with Cangxi Tongbi Capsules, si-circRNA＿0008365, si-NC, and Cangxi Tongbi Capsules combined with si-circRNA＿0008365. Flow cytometry and transmission electron microscopy were employed to determine the level of apoptosis and observe autophagosomes, respectively. Western blot was employed to reveal the changes in the protein levels of microtubule-associated protein light chain 3(LC3)Ⅱ/Ⅰ, Beclin-1, selective autophagy junction protein p62/sequestosome 1, collagen Ⅱ, a disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTS-5), and p38 MAPK. The mRNA levels of circRNA＿0008365, miR-1271, collagen Ⅱ, and ADAMTS-5 were determined by qRT-PCR. Hematoxylin-eosin staining was employed to reveal the pathological changes of the cartilage tissue of the knee, and enzyme-linked immunosorbent assay to measure the levels of interleukin-1β(IL-1β) and tumor necrosis factor-alpha(TNF-α). The chondrocytes treated with IL-1β showed down-regulated expression of circRNA＿0008365, up-regulated expression of miR-1271 and p38 MAPK, lowered autophagy level, increased apoptosis rate, and accelerated catabolism of extracellular matrix. The intervention with Cangxi Tongbi Capsules up-regulated the expression of circRNA＿0008365, down-regulated the expression of miR-1271 and p38 MAPK, increased the autophagy level, decreased the apoptosis rate, and weakened the catabolism of extracellular matrix. However, the effect of Cangxi Tongbi Capsules was suppressed after interfering with circRNA＿0008365. The in vivo experiments showed that Cangxi Tongbi Capsules dose-dependently inhibited the p38 MAPK pathway, enhanced chondrocyte autophagy, and mitigated articular cartilage damage and inflammatory response, thereby inhibiting the progression of KOA in rats. This study indicated that Cangxi Tongbi Capsules promoted chondrocyte autophagy by regulating the circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit the development of KOA.

Effects of diallyl trisulfide on cell cycle and apoptosis of human osteosarcoma cell line Saos-2 / 肿瘤

Objective: To investigate the effects of DATS (diallyl trisulfide) on cell proliferation, cell cycle and apoptosis of human osteosarcoma cell line Saos-2, and to explore their potential molecular mechanisms. Methods: Saos-2 cells were treated with 25, 50 and 100 μmol/L DATS for 24, 48, 72, and 96 h. The change of cell morphology was observed under an inverted microscope. The effect of DATS on the proliferation of Saos-2 cells was detected by MTT assay. After treatment with DATS at desired concentrations for various time intervals, the cell cycle and apoptotic rate of Saos-2 cells were analyzed by flow cytometry. The expressions of DATS-sensitive proteins in Saos-2 cells were systematically detected using 2-DE (two-dimensional gel electrophoresis) and mass spectrometry analysis. Results: DATS could inhibit the proliferation of Saos-2 cells in a dose- and time-dependent manner. Moreover, DATS could arrest the cell cycle at phase G0/G1 and induce the apoptosis in a time- and concentrationdependent manner. A total of 27 unique DATS-sensitive proteins, including 18 downregulated proteins and 9 upregulated proteins, were found in Saos-2 cells. Conclusion: DATS can suppress the proliferation of Saos-2 cells by blocking cell cycle progression and inducing apoptosis. The results of proteomics not only suggest the anticarcinogenic mechanism of DATS, but also provide a novel insight into the therapeutic targets for gene therapy of osteosarcoma. Copyright © 2013 by TUMOR.

Diagnosis of thyroid space-occupying lesions using real-time contrast-enhanced ultrasonography with sulphur hexafluoride microbubbles / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the value of contrast-enhanced ultrasound (CEUS) in the diagnosis of thyroid occupied lesions with injection of sulphur hexafluoride microbubbles.</p><p><b>METHODS</b>Fifty nine cases of conventional ultrasonic diagnosis of thyroid lesions in patients with 73 lesions re-sulfur hexafluoride microbubble ultrasound contrast real-time inspection, the use of CEUS and contrast pulse sequencing (CPS).</p><p><b>RESULTS</b>Seventy-three lesions were satisfied with the dynamic contrast perfusion imaging. Ultrasound contrast prompted the 15 lesions (13 cases) of malignant lesions by postoperative pathology confirmed papillary thyroid carcinoma, ultrasound images showed a low or mixed echo, boundary ambiguity, accompanied by Microcalcification foci, blood flow distribution I rank or grade III. Ultrasound prompted 58 lesions (46 cases) of benign lesions, blood flow distribution of grade II or III, the performance of a variety of two-dimensional ultrasound image, showing cystic, solid or liquid-solid mass, border clearance. Contrast with the surrounding thyroid tissue, lesions<or=20 mm in diameter papillary thyroid carcinoma are poor blood supply type; diameter>or=20 mm papillary thyroid carcinoma, manifested prior to the beginning of the adjacent thyroid substance to enhance and dissection, showing the form of high-enhanced perfusion; thyroid follicular adenoma as early as the beginning of the adjacent thyroid substance to enhance and dissection, mass retained within the contrast agent longer time, showing the form of high-enhanced perfusion; nodular goiter enhanced for thyroid nodules and adjacent synchronization started to pick up in real terms, after the 6 lesions showed nodular contrast slightly earlier than the adjacent thyroid substance started to pick up, 28 lesions with synchronous real beginning of the adjacent thyroid dissection, 11 lesions of nodular dissipated early on (started receding time < or = 25 s) adjacent to thyroid substance.</p><p><b>CONCLUSION</b>The real-time CEUS with injecting sulphur hexafluoride microbubbles is valuable in the diagnosis and differential diagnosis of the thyroid occupied lesions.</p>

Studies on Diffusion Properties of Alginate Gel Entrapped with Thiobacillus ferrooxidans Cells / 中国生物工程杂志

By non-steady method,the effective diffusivity of ferrous sulphate within alginate calcium gel entrapped without bacteria was measured.Meanwhile the oxidation ability of entrapped bacteria was analyzed.Experimental results showed that the effective diffusion coefficient of ferrous sulphate decreased with the increase of alginate concentration,the optimum alginate concentration is 2%(W/V).The effect of calcium chloride on the effective diffusivity was neglectable.The incubation of ferrooxidans would pass through 10 hours,and the diffusion coefficient within gel entrapped Thiobacillus ferrooxidans cells was less remarkably than that of ferrous sulphate without entrapped cells.For the entrapped cells,the absolute oxidation time was shortest and the rate change was fastest with the initial Fe concentration 5g/L.The absolute oxidation time was same when the initial Fe concentration was 8g/L and 10g/L.