Biological roles of microRNA-140 in tumor growth, migration, and metastasis of osteosarcoma in vivo and in vitro.

The objective of this study was to explore the biological roles of microRNA-140 (miR-140) in tumor growth, migration, and metastasis of osteosarcoma (OS) in vivo and in vitro. Between 2007 and 2014, 47 cases of OS samples and normal bone tissue samples adjacent to OS were selected from our hospital. Tissue biopsies from OS patients were used to measure miR-140 levels to obtain a correlation between clinicopathological features and miR-140 expression. In vitro, MG63 human osteosarcoma cells were divided into four groups: blank group, miR-140 mimic group, miR-140 inhibitor group, and negative control (NC; empty plasmid) group. qRT-PCR was used to detect miR-140 expression, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cell proliferation, flow cytometry was used to detect cell cycle distribution, and scratch migration assay was used to detect cell migration. In vivo, the relative expression of miR-140 level in OS tissue was lower than that in the adjacent normal bone tissue. miR-140 expression is inversely correlated with tumor size, Enneking stage, and tumor metastasis. In vitro, compared with blank group and NC group, relative miR-140 expression was increased, cell proliferation was inhibited, cell population in G0/G1 phase was increased, cell population in G2/M phase and S phases and proliferation index (PI), and cell migration distance were decreased in the miR-140 mimic group, but the relative expression and all the cell indexes were found opposite trend in the miR-140 inhibitor group. In conclusion, in vivo and vitro findings provided evidence that miR-140 could inhibit the growth, migration, and metastasis of OS cells.

Retraction Notice to: lncRNA CEBPA-AS1 Overexpression Inhibits Proliferation and Migration and Stimulates Apoptosis of OS Cells via Notch Signaling.

[This retracts the article DOI: 10.1016/j.omtn.2019.10.017.].

[Clinical observation on plastic splint treatment of middle clavicle fracture based on a new classification].

OBJECTIVE: According to 73 patients with middle clavicle fracture treated conservatively, a new classification of middle clavicle fracture was proposed, and the clinical effect of plastic splint in the treatment of middle clavicle fracture was observed. METHODS: Total 73 patients with middle clavicle fracture treated with plastic splint from September 2018 to August 2020 were analyzed retrospectively. All the patients were divided into 4 types according to the degree of fracture displacement. There were 16 cases of typeⅠ, including 7 males and 9 females, ranging in age from 18 to 37 years old, with a mean of (28.6±7.8) years old;12 cases of mild disease, 3 cases of moderate disease and 1 case of severe disease. There were 16 cases of type Ⅱ, including 6 males and 10 females, ranging in age from 25 to 49 years old, with a mean of (37.3±9.4) years old;5 cases of mild disease, 8 cases of moderate disease and 3 cases of severe disease. There were 7 cases of type Ⅲ, including 4 males and 3 females, ranging in age from 33 to 57 years old;2 cases of mild disease, 3 cases of moderate disease and 2 cases of severe disease. There were 34 cases of type Ⅳ, including 16 males and 18 females, ranging in age from 48 to 82 years old, with a mean of(66.4±14.9) years old;7 cases of mild disease, 17 cases of moderate disease and 10 cases of severe disease. All patients received plastic splint external fixation for 4 weeks. Visual analgue scale (VAS) and Constant-Murley shoulder scores before treatment and 1, 3 and 9 months after treatment were observed and recorded to evaluate the change of pain degree and shoulder function recovery before and after treatment. The patients' satisfaction with the appearance after treatment was recorded at the latest follow-up. The X-ray findings at the latest follow-up were used to judge whether the patient had fracture nonunion. And according to the fracture healing time and imaging findings, the excellent and good rate of clinical curative effect in patients with different types was obtained. RESULTS: All patients were followed up, and the duration ranged from 9 to 11 months, with a mean of (9.8±0.7) months. The VAS scores of typeⅠ, typeⅡand type Ⅳ before treatment were 2.88±0.83, 3.67±0.80 and 6.92±1.71 respectively, which were decreased to 0.54±0.19, 0.77±0.25 and 1.18±0.17 respectively after 9 months of treatment. The Constant-Murley shoulder scores of typeⅠ, typeⅡand type Ⅳ were 65.81±2.09, 63.50±2.22 and 47.93±2.91 respectively before treatment, and increased to 88.56±2.11, 85.12±2.23 and 71.25±2.16 respectively after 9 months of treatment. Five patients were not satisfied with the appearance after treatment;6 patients had no obvious continuous callus passing through after 9 months of treatment, which was fracture nonunion. CONCLUSION: The classification of middle clavicle fracture is more appropriate to the clinic, which has a certain clinical guiding significance for the selection of treatment methods and prognosis of middle clavicle fracture. Plastic splint is effective in the treatment of middle clavicle fracture without obvious displacement and overlapping displacement, and the incidence of complications is low. It can be popularized in clinic.

Therapeutic potential of experimental autoimmune encephalomyelitis by Fasudil, a Rho kinase inhibitor.

The migration of aberrant inflammatory cells into the central nervous system plays an important role in the pathogenesis of demyelinating diseases potentially through the Rho/Rho-kinase (Rock) pathway, but direct evidence from human and animal models remains inadequate. Here we further confirm that Fasudil, a selective Rock inhibitor, has therapeutic potential in a mouse model of myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE). The results show that Fasudil decreased the development of EAE in C57BL/6 mice. Immunohistochemistry disclosed that expression of Rock-II in the perivascular spaces and vascular endothelial cells of spleens, spinal cords, and brains was elevated in EAE and was inhibited in the Fasudil-treated group. T-cell proliferation specific to MOG(35-55) was markedly reduced, together with a significant down-regulation of interleukin (IL)-17, IL-6, and MCP-1. In contrast, secretion of IL-4 was increased, and IL-10 was slightly elevated. There were no differences in the percentages of CD4(+)CD25(+), CD8(+)CD28(-), and CD8(+)CD122(+) in mononuclear cells. Histological staining disclosed a marked decrease of inflammatory cells in spinal cord and brain of Fasudil-treated mice. These results, together with previous studies showing the inhibitory effect of Fasudil on T-cell migration, might expand its clinical application as a new therapy for multiple sclerosis by decreasing cell migration and regulating immune balance.

[Characteristics of clinical findings and radiological assessment of high grade developmental spondylolisthesis].

OBJECTIVES: To investigate clinical appearance and radiological characteristics of high grade developmental spondylolisthesis and their significance. METHODS: In a retrospective study, a group of 6 female patients, who were diagnosed as high grade developmental spondylolisthesis and treated in Peking University third hospital from March 2007 to December 2008 were included. Clinical and radiological characteristics of the 6 patients were investigated and the following parameters were measured on standing lateral X-ray: PI (pelvic incidence), SK (sacral kyphosis) and LL (Lumbar lordosis). A series of 44 patients who came to out-patient department due to LBP and had no positive findings on lateral lumbar X-ray were selected as the control group. The four parameters were compared between study group and control group. Clinical meanings of significant difference were discussed. RESULTS: Clinical findings of high grade developmental spondylolisthesis included bending of knees, deformity of trunk and sciatica. Radiological appearances were characterized with kyphosis of lumbo-sacral joint, retroverted pelvis and domed sacrum. Spondylolisthesis patients has an average PI of (52 +/- 7) degrees which was significant higher than the control group [(43 +/- 8) degrees ] (P < 0.01). LL of study group [(51 +/- 10) degrees ] was higher than that of the control group [(18 +/- 9) degrees ] (P < 0.01) and SK of the study group [(12 +/- 11) degrees ] were lower than that of the control group [(21 +/- 10) degrees ] (P < 0.05). CONCLUSIONS: Characteristics of clinical findings of spondylolisthesis patients have obvious cosmetic appearance, significant pain and lower neurological deficits. Radiological characteristics display deformity of lumbo-sacral joint. PI, LL and SK are significant parameters for high grade spondylolisthesis.

lncRNA CEBPA-AS1 Overexpression Inhibits Proliferation and Migration and Stimulates Apoptosis of OS Cells via Notch Signaling.

Osteosarcoma (OS) is the most common primary bone malignancy derived from primitive bone-forming mesenchymal cells. Long noncoding RNA (lncRNA) expression profiles have been intensively studied for their involvement in OS. Herein, we clarify whether lncRNA CEBPA-AS1 is a regulator of NCOR2 in OS cells. Microarray-based expression analysis identified OS-related differentially expressed lncRNA and predicted microRNAs (miRs) binding to lncRNA and mRNA. lncRNA CEBPA-AS1 and NCOR2 were found to be weakly expressed in OS tissues and cells. Next, functional investigation revealed that lncRNAs CEBPA-AS1 bound to miR-10b-5p to upregulate NCOR2. Following that, gene-targeted knockdown and overexpressed recombinant vectors of lncRNA CEBPA-AS1 and NCOR2 were constructed to explore the effects of lncRNA CEBPA-AS1 and NCOR2 on cell proliferation, differentiation, migration, and apoptosis. Finally, tumor formation was measured in nude mice. lncRNA CEBPA-AS1 overexpression or NCOR2 elevation inhibited cell proliferation and migration, and alkaline phosphatase (ALP) and bone gla protein (BGP) activity, while enhancing apoptosis and tumor formation. Furthermore, NCOR2 was elevated in response to lncRNA CEBPA-AS1 overexpression, thus repressing the Notch signaling pathway. Taken together, lncRNA CEBPA-AS1 overexpression inhibits OS progression through diminishing activation of the Notch signaling pathway via upregulating NCOR2. Therefore, lncRNA CEBPA-AS1 may serve as a molecular target for treating OS.

Two new mixed-ligand coordination polymers based on multi-N chelating ligand inhibit YAP expression and induce caspase-mediated spinal tumor cell apoptosis.

Two new coordination polymers [Zn (bdc)(bpybzimH2)](DMF)0.5 (1, H2bdc=1,4-dicarboxybenzene, bpybzimH2=6,6'-bis-(1H-benzoimidazol-2-yl)-2,2'-bipyridine, DMF=N,N-dimethylformamide) and [Co (bpybzimH2)(sbc)]H2O (2, H2sbc=4-mercaptobenzoic acid) have been successfully prepared under solvothermal conditions using the multi-N chelating organic ligand bpybzimH2 as the foundational building block. In addition, the Cell Counting Kit-8 assay was conducted to evaluate the anti-proliferation activity of compounds 1 and 2 against human spinal tumor cells OPM-2. The cell viability curves showed that the two compounds have anti-proliferation activity on spinal tumor cells, and the activity of compound 1 is higher than compound 2. The annexin V-FITC/PI assay and western blot were used to detect the apoptotic percentage of OPM-2 cells incubated with compounds 1 and 2. The YAP protein expression and its role in cell apoptosis were further studied with qRT-PCR, immunoblotting, and flow cytometer.

Mulberrin (Mul) reduces spinal cord injury (SCI)-induced apoptosis, inflammation and oxidative stress in rats via miroRNA-337 by targeting Nrf-2.

Spinal cord injury (SCI) is a major reason of paralysis, disability and even death in severe cases. Mulberrin (Mul) is a major compound of ramulus mori and turned out that it has potent ability on tyrosinase inhibition. Ramulus mori has been reported to possess anti-inflammation, anti-oxidative stress and anti-apoptosis effects. However, the effects of Mul on SCI progression and the underlying molecular mechanism have never been elucidated before. Here, we established a SCI rat model, which subsequently received Mul treatment. The findings showed that Mul treatment improved the functional recovery of SCI rats, along with reduced spinal cord water contents and myeloperoxidase (MPO) activity. SCI-induced apoptosis was attenuated by Mul, as evidenced by the reduced TUNEL-positive cells, and the decreased pro-apoptotic signals expressions, while increased anti-apoptotic molecule. Further, Mul inhibited inflammatory response in the spinal cord tissues of SCI rats through inactivating nuclear factor-kappa B (NF-κB) pathway. Oxidative stress was also decreased by Mul treatment in SCI mice, associated with the up-regulation of heme oxygenase-1 (HO-1)/nuclear factor E2-related factor 2 (HO-1/Nrf-2) pathway. Significantly, SCI rats showed high expression of miR-337 in spinal cord tissue samples. Astrocytes (AST) stimulated by LPS also had higher expression of miR-337. Nrf-2 was found to be a direct target for miR-337. Over-expressing miR-337 markedly reduced Nrf-2 expression in AST, whereas inhibiting miR-337, Nrf-2 expressions were significantly increased. In vitro, AST transfected with miR-337 inhibitor showed attenuated inflammation, apoptosis and oxidative stress in LPS-treated AST, which was, intriguingly, abolished by Nrf-2 knockdown. Together, the findings above indicated that Mul could attenuate SCI by reducing miR-337 expressions to reduce apoptosis, inflammation and oxidative stress via regulating Nrf-2.

Two new mixed-ligand coordination polymers based on multi-N chelating ligand inhibit YAP expression and induce caspase-mediated spinal tumor cell apoptosis

Two new coordination polymers [Zn (bdc)(bpybzimH2)](DMF)0.5 (1, H2bdc=1,4-dicarboxybenzene, bpybzimH2=6,6′-bis-(1H-benzoimidazol-2-yl)-2,2′-bipyridine, DMF=N,N-dimethylformamide) and [Co (bpybzimH2)(sbc)]H2O (2, H2sbc=4-mercaptobenzoic acid) have been successfully prepared under solvothermal conditions using the multi-N chelating organic ligand bpybzimH2 as the foundational building block. In addition, the Cell Counting Kit-8 assay was conducted to evaluate the anti-proliferation activity of compounds 1 and 2 against human spinal tumor cells OPM-2. The cell viability curves showed that the two compounds have anti-proliferation activity on spinal tumor cells, and the activity of compound 1 is higher than compound 2. The annexin V-FITC/PI assay and western blot were used to detect the apoptotic percentage of OPM-2 cells incubated with compounds 1 and 2. The YAP protein expression and its role in cell apoptosis were further studied with qRT-PCR, immunoblotting, and flow cytometer.

Ani-survivin DNAzymes inhibit cell proliferation and migration in breast cancer cell line MCF-7.

Survivin, a new member of the inhibitor of apoptosis protein (IAP) family, both inhibits apoptosis and regulates the cell cycle. It is overexpressed in breast tumor tissues. In this study, we designed two survivin specific DNAzymes (DRz1 and DRz2) targeting survivin mRNA. The results showed that DRz1 could decrease the expression of survivin by nearly 60%. Furthermore, DRz1 significantly inhibited cell proliferation, induced apoptosis and inhibited migration in MCF-7 cells. In addition, down-regulation of survivin expression was associated with increased caspase-3 and -9 activities in MCF-7 cells after 24 h transfection. In our experiments, the efficacy of DRz1 to influence survivin levels and associated effects were better than DRz2. Survivin-DRz1 might have anti-tumorigenic activity and may potentially provide the basis for a novel therapeutic intervention in breast cancer treatment.