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Objective：To investigate the effect of miR⁃27b⁃3p on osteosarcoma cells growth and its potential molecular mechanism. Methods：①The expression of miR⁃27b⁃3p in osteosarcoma cell lines was analyzed by RT⁃qPCR. ②CCK⁃8 assay, colony formation assay and flow cytometry analysis were used to detect the effect of miR⁃27b⁃3p and structure⁃specific recognition protein⁃1（SSRP1）on osteosarcoma cell growth. ③The miRNA target prediction database was used to predict potential target gene of miR⁃27b⁃3p, and dual luciferase reporter assay was performed to demonstrate the relationship of miR⁃27b⁃3p and its target genes. ④Efficiency of knockdown or overexpression was validated by Western blot analysis. Results：①The expression of miR ⁃27b ⁃3p in osteosarcoma cell lines and clinical samples was low, and overexpression of miR⁃27b⁃3p inhibited the growth of osteosarcoma cells. ②Bioinformatics analysis and dual⁃fluorescence reporter assay confirmed that SSRP1 was a target gene of miR⁃27b⁃3p, and the expression of SSRP1 was regulated by miR⁃27b⁃3p. ③Silencing of SSRP1 significantly inhibited cell proliferation and increased cell apoptosis of osteosarcoma cells. ④Overexpression of SSRP1 partly reversed the inhibitory effect of miR⁃27b⁃3p on osteosarcoma cell growth. Conclusion：miR⁃27b⁃3p is low expressed in osteosarcoma cells, and it regulates the growth of osteosarcoma cells by targeting SSRP1. Targeting the miR⁃27b⁃3p/ SSRP1 axis may become a new therapeutic strategy for the treatment of osteosarcoma. [ABSTRACT FROM AUTHOR]

Objective To investigate the serum expression characteristics of miR-27b-3p and miR-342-3p in type 2 diabetic nephropathy (DN) patients and the diagnostic value for DN. Methods A total of 157 patients with DN were selected and divided into the normal albuminuria group (NA group, n=49), the microalbuminuria group (MA group, n=41) and the macroalbuminuria group (DN group, n=67) according to the ratio of albumin to creatinine (UACR) of urine specimens. Clinical data and the expression of serum miR-27b-3p and miR-342-3p, renal function and urinary fibrosis indicators were detected by real-time quantitative polymerase chain reaction. Blood urea nitrogen (BUN) and blood creatinine (Scr), were detected by automatic biochemical analyzer. Urine type Ⅳ collagen (Ⅳ-C), laminin (LN) and type Ⅲ procollagen peptide (PCⅢ) were detected by double-antibody sandwich enzyme-linked immunosorbent assay. The factors influencing the pathogenesis of DN and the diagnostic value of miR-27b-3p and miR-342-3p for DN were analyzed. Results Compared with the NA group, the levels of HbA1c, UACR, urinary Ⅳ-C, urinary LN and urinary PCⅢ were increased, while the expresson levels of miR-27b-3p and miR-342-3p were decreased in the DN group and the MA group (P＜0.05). Levels of BUN and Scr were increased in the DN group (P＜0.05), and eGFR level decreased (P＜0.05). The higher level of urinary PCⅢ was a risk factor for DN, while the higher levels of miR-27b-3p, miR-342-3p and eGFR were protective factors of DN (P＜0.05). The expressions levels of miR-27b-3p and miR-342-3p were positively correlated with eGFR (r = 0.647, 0.661, P＜0.05) and negatively correlated with urinary PCⅢ (r=-0.739, P＜0.05). The area under the curve for diagnosing DN by combining eGFR, urinary PCⅢ, miR-27b-3p and miR-342-3p was higher than that of eGFR, urinary PCⅢ, miR-27b-3p, miR-342-3p alone (Z=5.631, 5.794, 6.715, 4.577, P＜0.05). Conclusion The decreased expressions of serum miR-27b-3p and miR-342-3p are associated with decreased renal function and DN occurrence in patients with type 2 diabetes, which can be used as biological indicators for the auxiliary diagnosis of DN. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of circRNA&#95;005647 on fibrotic phenotype of mouse cardiac fibroblasts (CFs) and explore its mechanism., Methods: We used an angiotensin Ⅱ (Ang-Ⅱ) capsule pump (daily dose of 1.46 mg/kg for 2 weeks) to establish a mouse model of myocardial fibrosis in C57BL/6 mice. Masson staining was used to detect myocardial fibrosis in the myocardium. The expression of circRNA&#95;005647 in the myocardium of Ang-Ⅱ-infused mice and in Ang-Ⅱ-treated CFs were detected with real-time PCR. Actinomycin D and RNase R exonuclease digestion were used to test the stability of circRNA&#95;005647 in mouse CFs. Over- expression of circRNA&#95;005647 was achieved in the CFs by infecting the cells with a recombinant circRNA&#95;005647 adenovirus (rAd-circRNA&#95;005647), and the expressions of Col1a1, Col3a1 and Acta2 were detected in the cells with real-time PCR and Western blotting. Dual luciferase reporter assay, RNA antisense purification and RNA Pull down assay were performed to identify the interaction between circRNA&#95;005647 and miR-27b-3p., Results: CircRNA&#95;005647 was up- regulated in the myocardium of Ang-Ⅱ- infused mice and in Ang-Ⅱ-treated mouse CFs ( P < 0.01). CircRNA&#95;005647 was more stable than its host gene Myosin IXA ( Myo9a ) in response to actinomycin D ( P < 0.01) and RNase R exonuclease treatment. The expressions of fibrosis-associated genes was down-regulated in the CFs over-expressing circRNA&#95; 005647 ( P < 0.05). Dual luciferase reporter assay, RNA antisense purification and RNA Pull down assay revealed the interaction between miR-27b-3p and circRNA&#95;005647. MiR-27b-3p obviously enhanced the fibrotic phenotype but reversed the inhibitory effect of circRNA&#95;005647 on the expression of fibrosis associated genes in the CFs ( P < 0.05)., Conclusions: CircRNA&#95;005647 is upregulated in cardiac fibrosis and inhibits the expression of fibrosis-related genes through sponging miR-27b-3p in mouse CFs.