MicroRNA-493-5p inhibits proliferation and metastasis of osteosarcoma cells by targeting Kruppel-like factor 5.

Osteosarcoma, including spinal osteosarcoma, has properties of high degree of malignancy, high rate of recurrence, and high incidence of metastasis. microRNAs can exert oncogenic or tumor suppressive roles in cancer cells. This study explored the effects of microRNA-493-5p (miR-493-5p) on osteosarcoma cell viability, migration, invasion, and apoptosis, as well as the underlying possible mechanism. First, the expression of miR-493-5p in osteosarcoma tissues and cells was detected using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Then, the effects of miR-493-5p overexpression (or suppression) on osteosarcoma cell viability, migration, invasion, and apoptosis, as well as Kruppel-like factor 5 (KLF5) expression, were assessed using the Cell Counting Kit-8 assay, two-chamber transwell assay, Annexin V-FITC/PI apoptosis detection kit, qRT-PCR, and western blotting, respectively. Finally, the roles of KLF5 in miR-493-5p suppression-induced U20S cell viability, migration, and invasion enhancement, as well as the PI3K/AKT pathway activation, were evaluated. We found that miR-493-5p had lower expression in tumor tissues of spinal osteosarcoma and osteosarcoma cells. Overexpression of miR-493-5p inhibited osteosarcoma U20S cell viability, migration, and invasion, but induced cell apoptosis. On the contrary, suppression of miR-493-5p-promoted U20S cell viability, migration, and invasion. KLF5 was a direct target gene of miR-493-5p, which participated in the effects of miR-493-5p on U20S cell viability, migration, invasion, and apoptosis. Furthermore, suppression of the miR-493-5p activated PI3K/AKT pathway in U20S cells by upregulating KLF5. In conclusion, we revealed that miR-493-5p exerted tumor suppressive roles in spinal osteosarcoma and osteosarcoma cells. Overexpression of miR-493-5p inhibited proliferation and metastasis of osteosarcoma cells by downregulating KLF5 and inactivating the PI3K/AKT signaling pathway.

The Effects of Post Heat Treatment on the Microstructural and Mechanical Properties of an Additive-Manufactured Porous Titanium Alloy.

In this work, Ti-6Al-4V (Ti64) porous structures were prepared by selective laser melting (SLM), and the effects of post heat treatment on its microstructural and mechanical properties were investigated. The results showed that as SLM samples were mainly composed of needle-like α' martensite. Heat treatment at 750 °C caused α' phase to decompose, forming a lamellar α+ß mixed microstructure. As the heat treatment temperature increased to 950 °C, the width of lamellar α phase gradually increased to 3.1 µm. Heat treatment also reduced the compressive strength of the samples; however, it significantly improved the ductility of the porous Ti64. Moreover, heat treatment improved the energy absorption efficiency of the porous Ti64. The samples heat-treated at 750 °C had the highest energy absorption of 233.6 ± 1.5 MJ/m3 at ε = 50%.

miR-342-5p inhibits expression of Bmp7 to regulate proliferation, differentiation and migration of osteoblasts.

BACKGROUND: Fracture healing is a complex process, and patients with fracture will undergo non-union or compromised regeneration. MicroRNA (miR)-342-5p is a Notch downstream molecule, and its roles in fracture healing remain unclear. We aimed to explore the functional roles of miR-342-5p in osteoblasts as well as the underlying mechanisms. METHODS: The expression of miR-342-5p in differentiation of MC3T3-E1 cells or hMSCs was examined by quantitative reverse transcription PCR (qRT-PCR). The effects of aberrantly expressed miR-342-5p on cell proliferation, apoptosis, migration, and expressions of proteins associated with proliferation and osteogenic differentiation were determined by Cell Counting Kit-8, trypan blue staining, flow cytometry, Transwell assay, Western blot and qRT-PCR assays, respectively. The downstream factor and the target genes of miR-342-5p as well as the involvements of the mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) pathway were finally assessed. RESULTS: miR-342-5p level was decreased during differentiation of MC3T3-E1 cells or hMSCs. After cell transfection, miR-342-5p overexpression significantly reduced cell viability, induced apoptosis, inhibited proliferation, migration and differentiation, and down-regulated Bmp7 expression. Subsequent experiments showed the effects of miR-342-5p inhibition on MC3T3-E1 cells were abrogated by Bmp7 knockdown. Additionally, COL4A6 and Bmp2 were predicated as target genes of miR-342-5p. Finally, phosphorylated levels of MEK and ERK were increased by miR-342-5p inhibition via up-regulating Bmp7 expression. CONCLUSION: miR-342-5p inhibition promoted proliferation, migration and differentiation of osteoblasts via regulating Bmp7, along with activation of the MEK/ERK pathway.

Green tea polyphenols protect PC12 cells against H2O2-induced damages by upregulating lncRNA MALAT1.

It is of significance to alleviate oxidative damages for the treatment of spinal cord injury (SCI). Studies have ascertained that green tea polyphenols (GTPs) exert protective activities against oxidative damages. In this study, we aimed to investigate the protective effects of GTP against H2O2-caused injuries in PC12 cells as well as the molecular underpinnings associated with long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1). PC12 cells were preincubated with GTP prior to H2O2 stimulation. Furthermore, MALAT1-deficient PC12 cells were constructed by transfection and identified by quantitative real-time polymerase chain reaction (qRT-PCR) assay. Next, viability and apoptosis were detected by cell counting kit-8 and flow cytometry, respectively. Meanwhile, Western blot assay was carried out to monitor the expression alteration of proteins associated with apoptosis (Bcl-2, Bax, pro-Caspase-3/9, and cleaved Caspase-3/9) and autophagy (microtubule-associated protein 1 light chain 3 (LC3)-II, LC3-I, Beclin-1, and p62). Moreover, we examined the expression of ß-catenin and dissected the phosphorylation of phosphatidylinositol 3'-kinase (PI3K) and protein kinase B (AKT). We found that H2O2 decreased the viability of PC12 cells while initiated apoptosis and autophagy processes. GTP-preincubated PC12 cells maintained the viability and resisted the apoptosis and autophagy induced by H2O2. Pointedly, GTP-pretreated PC12 cells showed an increase in MALAT1 after H2O2 stimulation. Of note, the protective effects of GTP were buffered in MALAT1-deficient cells in response to H2O2. The expression of ß-catenin and phosphorylation of PI3K and AKT were upregulated by GTP, while MALAT1 knockdown led to opposite results. To sum up, GTP protected PC12 cells from H2O2-induced damages by the upregulation of MALAT1. This process might be through activating Wnt/ß-catenin and PI3K/AKT signal pathways.

Kaempferol protects chondrogenic ATDC5 cells against inflammatory injury triggered by lipopolysaccharide through down-regulating miR-146a.

Kaempferol is a kind of bioflavonoid exerts diverse pharmacological activities, including anti-apoptotic and anti-inflammatory activities. Kaempferol has been recognized as an effective agent for alleviating the clinical symptoms of osteoarthritis (OA). This study aimed to provide evidence that Kaempferol has potential in the management of OA. Lipopolysaccharide (LPS) stimulation induced a significant cell death and inflammatory injury in ATDC5 cells, as evidenced by the decreased cell viability, the induced apoptosis, the activated caspase-3, and the excessive production of IL-6, IL-8 and TNF-α. Precondition of cells with Kaempferol prevented apoptosis and the release of proinflammatory cytokines triggered by LPS. miR-146a was down-regulated by Kaempferol treatment, and Decorin was up-regulated by miR-146a overexpression. Consistently, both silence of miR-146a and Decorin exhibited Kaempferol-like effects towards ATDC5 cells stimulated by LPS. Moreover, Decorin silence activated PI3K/AKT/mTOR signaling pathway. In rat model of OA, the expression of miR-146a and Decorin in cartilage tissues was repressed by Kaempferol. Also, the activated PI3K/AKT/mTOR signaling pathway in OA animal model was enhanced by Kaempferol administration. These data suggested that Kaempferol exerted potential anti-OA effects through down-regulation of miR-146a, and thus repressing the expression of Decorin.

Poly[µ(2)-aqua-µ(4)-naphthalene-1,8-dicarboxyl-ato-manganese(II)].

The asymmetric unit of the title complex, [Mn(C(12)H(6)O(4))(H(2)O)](n), contains one Mn(II) ion, one 1,8-naphthalene-dicarboxyl-ate (1,8-NDC) ligand and one water mol-ecule. The Mn(II) ion is six-coordinated within a distorted octa-hedral coordination geometry, in which the equatorial sites are occupied by four carboxyl-ate O atoms from four different 1,8-NDC ligands, while the axial positions are occupied by two O atoms of two coordinated water mol-ecules. Adjacent Mn(II) centres are bridged by one coordinated water and two carboxyl-ate groups in a syn-syn mode to form infinite chains along the b axis, which are further cross-linked by the naphthalene spacers of the 1,8-NDC ligands to produce a two-dimensional extended network.

DCDB 2.0: a major update of the drug combination database.

Experience in clinical practice and research in systems pharmacology suggested the limitations of the current one-drug-one-target paradigm in new drug discovery. Single-target drugs may not always produce desired physiological effects on the entire biological system, even if they have successfully regulated the activities of their designated targets. On the other hand, multicomponent therapy, in which two or more agents simultaneously interact with multiple targets, has attracted growing attention. Many drug combinations consisting of multiple agents have already entered clinical practice, especially in treating complex and refractory diseases. Drug combination database (DCDB), launched in 2010, is the first available database that collects and organizes information on drug combinations, with an aim to facilitate systems-oriented new drug discovery. Here, we report the second major release of DCDB (Version 2.0), which includes 866 new drug combinations (1363 in total), consisting of 904 distinctive components. These drug combinations are curated from ∼140,000 clinical studies and the food and drug administration (FDA) electronic orange book. In this update, DCDB collects 237 unsuccessful drug combinations, which may provide a contrast for systematic discovery of the patterns in successful drug combinations. Database URL: http://www.cls.zju.edu.cn/dcdb/