miR-128 Targets the SIRT1/ROS/DR5 Pathway to Sensitize Colorectal Cancer to TRAIL-Induced Apoptosis.

BACKGROUND/AIMS: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is an ideal anti-tumor drug because it exhibits selective cytotoxicity against cancer cells. However, certain cancer cells are resistant to TRAIL, and the potential mechanisms are still unclear. The aim of this study was to reduce the resistance of colorectal cancer (CRC) cells to TRAIL. METHODS: Quantitative real-time PCR analysis was performed to detect the expression of microRNA-128 (miR-128) in tissues from patients with CRC and CRC cell lines. MTT assays were used to evaluate the effect of miR-128 on TRAIL-induced cytotoxicity against CRC cell lines. The distribution of death receptor 5 (DR5) and the production of reactive oxygen species (ROS) were detected by flow cytometry analysis. Western blot, flow cytometry, and luciferase reporter assays were performed to evaluate the potential mechanism and pathway of miR-128-promoted apoptosis in TRAIL-treated CRC cells. RESULTS: MiR-128 expression was downregulated in tumor tissues from patients with CRC as well as in CRC cell lines in vitro. The enforced expression of miR-128 sensitized CRC cells to TRAIL-induced cytotoxicity by inducing apoptosis. Mechanistically, bioinformatics, western blot analysis, and luciferase reporter assays showed that miR-128 directly targeted sirtuin 1 (SIRT1) in CRC cells. miR-128 overexpression suppressed SIRT1 expression, which promoted the production of ROS in TRAIL-treated CRC cells. This increase of ROS subsequently induced DR5 expression, and thus increased TRAIL-induced apoptosis in CRC cells. CONCLUSION: The combination of miR-128 with TRAIL may represent a novel approach for the treatment of CRC.

miR-142-3p Suppresses Cell Growth by Targeting CDK4 in Colorectal Cancer.

BACKGROUND/AIMS: Deregulation of microRNAs (miRNAs) has been associated with a variety of cancers, including colorectal cancer (CRC). Here, we investigated anomalous miR-142-3p expression and its possible functional consequences in primary CRC samples. METHODS: The expression of miR-142-3p was measured by quantitative RT-PCR in 116 primary CRC tissues and adjacent non-tumor tissues. The effect of miR-142-3p up- or down-regulation in CRC-derived cells was evaluated in vitro by cell viability and colony formation assays and in vivo by growth assays in xenografted nude mice. RESULTS: Using quantitative RT-PCR, we found that miR-142-3p was down-regulated in 78.4 % (91/116) of the primary CRC tissues tested when compared to the adjacent non-tumor tissues. We also found that the miR-142-3p mimic reduced in vitro cell viability and colony formation by inducing cell cycle arrest in CRC-derived cells, and inhibited in vivo tumor cell growth in xenografted nude mice. Inversely, we found that the miR-142-3p inhibitor increased the viability and colony forming capacity of CRC-derived cells and tumor cell growth in xenografted nude mice. In addition, we identified CDK4 as a potential target of miR-142-3p by predictions and dual-luciferase reporter assays. Concordantly, we found that miR-142-3p mimics and inhibitors could decrease and increase CDK4 protein levels in CRC-derived cells, respectively. CONCLUSION: From our results we conclude that miR-142-3p may act as a tumor suppressor in CRC and may serve as a tool for miRNA-based CRC therapy.

Knockdown of Long Non-Coding RNA XIST Inhibited Doxorubicin Resistance in Colorectal Cancer by Upregulation of miR-124 and Downregulation of SGK1.

BACKGROUND/AIMS: Doxorubicin (DOX) is a widely used chemotherapeutic agent for colorectal cancer (CRC). However, the acquirement of DOX resistance limits its clinical application for cancer therapy. Mounting evidence has suggested that aberrantly expressed lncRNAs contribute to drug resistance of various tumors. Our study aimed to explore the role and molecular mechanisms of lncRNA X-inactive specific transcript (XIST) in chemoresistance of CRC to DOX. METHODS: The expressions of XIST, miR-124, serum and glucocorticoid-inducible kinase 1 (SGK1) mRNA in DOX-resistant CRC tissues and cells were detected by qRT-PCR or western blot analysis. DOX sensitivity was assessed by detecting IC50 value of DOX, the protein levels of P-glycoprotein (P-gp) and glutathione S-transferase-π (GST-π) and apoptosis. The interactions between XIST, miR-124 and SGK1 were confirmed by luciferase reporter assay, qRT-PCR and western blot. Xenograft tumor assay was used to verify the role of XIST in DOX resistance in CRC in vivo. RESULTS: XIST expression was upregulated and miR-124 expression was downregulated in DOX-resistant CRC tissues and cells. Knockdown of XIST inhibited DOX resistance of CRC cells, as evidenced by the reduced IC50 value of DOX, decreased P-gp and GST-π levels and enhanced apoptosis in XIST-silenced DOX-resistant CRC cells. Additionally, XIST positively regulated SGK1 expression by interacting with miR-124 in DOX-resistant CRC cells. miR-124 suppression strikingly reversed XIST-knockdown-mediated repression on DOX resistance in DOX-resistant CRC cells. Moreover, SGK1-depletion-elicited decrease of DOX resistance was greatly restored by XIST overexpression or miR-124 inhibition in DOX-resistant CRC cells. Furthermore, XIST knockdown enhanced the anti-tumor effect of DOX in CRC in vivo. CONCLUSION: XIST exerted regulatory function in resistance of DOX possibly through miR-124/SGK1 axis, shedding new light on developing promising therapeutic strategy to overcome chemoresistance in CRC patients.

Admission Inflammation Markers Influence Long-term Mortality in Elderly Patients Undergoing Hip Fracture Surgery: A Retrospective Cohort Study.

OBJECTIVES: Hip fractures in elderly patients are associated with a high mortality rate. Most deaths associated with hip fracture result from complications after surgery. Recent studies suggest that some inflammation biomarkers may be useful to estimate excess mortality. This study aimed to investigate the prognostic value of admission inflammation biomarkers in elderly patients with hip fracture. METHODS: We reports on a retrospective study of elderly hip fracture patients admitted to a hospital in China between January 2015 and December 2019. A total of 1085 patients were included in the study, and their demographic and pre-operative characteristics were analyzed. The inflammation biomarkers included monocyte to lymphocyte ratio (MLR), neutrophil to lymphocyte ratio (NLR), and C-reactive protein (CRP) to albumin ratio (CAR). The predictive performance of NLR, MLR and CAR was assessed by receiver operating characteristics (ROC) curve analysis and the association between admission inflammation markers and mortality was evaluated by Cox proportional regression. RESULTS: The 30-day, 1-year, 2-year, and 4-year mortality were 1.6%, 11.5%, 21.4% and 48.9%, respectively. The optimal cut-off values of admission NLR, MLR and CAR for 1-year mortality were 7.28, 0.76, and 1.36. After adjusting the covariates, preoperative NLR ≥ 7.28 (HR = 1.419, 95% CI: 1.080-1.864, p = 0.012) were found to be only independent risk factors with 4-year all-cause mortality, the preoperative CAR ≥ 1.36 was independently associated with 1-year (HR = 1.700, 95% CI: 1.173-2.465, p = 0.005), 2 year (HR = 1.464, 95% CI: 1.107-1.936, p = 0.008), and 4-year (HR = 1.341, 95% CI: 1.057-1.700, p = 0.016) all-cause mortality, While age, CCI score, and low hemoglobin at admission were also risk factors for postoperative all-cause mortality. CONCLUSION: Admission CAR and NLR may be useful indicators for predicting the long-term mortality of elderly patients undergoing hip fracture surgery, and that more research is needed to validate these findings.

Autologous grafts of double-strut fibular cortical bone plate to treat the fractures and defects of distal femur: a case report and review of literature.

We reported a 23-year-old man who was involved in a high-speed motorcycle accident. He sustained a closed fracture at the right distal femur. The primary fracture happened on February 2008. He underwent open reduction and internal fixation with cloverleaf plate. And one hundred days after the surgery, the proximal screws were pulled-out, but the bone union was not achieved. Treatment consisted of exchanging the cloverleaf plate with a locking compression plate and using an auto-iliac bone graft to fill the nonunion gap. In July 2009, the patient had a sharp pain in the right lower limb. The X-ray revealed that the plate implanted last year was broken, causing a nonunion at the fracture site. Immediately the plate and screws were removed and an intramedullary nail was inserted reversely from the distal femur as well as a 7 cm long bone from the right fibula was extracted and longitudinally split into two pieces to construct cortical bone plates. Then we placed them laterally and medially to fracture site, drilled two holes respectively, and fastened them with suture. We carried on auto-iliac bone grafting with the nonunion bone grafts. The follow-up at 15 months after operation showed that the treatment was successful, X-ray confirmed that there was no rotation and no angular or short deformity. We briefly reviewed the literature regarding such an unusual presentation and discussed in details the possible etiology and the advantages of autologous double-strut fibular grafts to cope with such an intractable situation.

A robust and efficient framework for tubular structure segmentation in chest CT images.

BACKGROUND: Tubular structure segmentation in chest CT images can reduce false positives (FPs) dramatically and improve the performance of nodules malignancy levels classification. OBJECTIVE: In this study, we present a framework that can segment the pulmonary tubular structure regions robustly and efficiently. METHODS: Firstly, we formulate a global tubular structure identification model based on Frangi filter. The model can recognize irregular vascular structures including bifurcation, small vessel, and junction, robustly and sensitively in 2D images. In addition, to segment the vessels from JVN, we design a local tubular structure identification model with a sliding window. Finally, we propose a multi-view voxel discriminating scheme on the basis of the previous two models. This scheme reduces the computational complexity of obtaining high entropy spatial tubular structure information. RESULTS: Experimental results have shown that the proposed framework achieves TPR of 85.79%, FPR of 24.83%, and ACC of 84.47% with the average elapsed time of 162.9 seconds. CONCLUSIONS: The framework provides an automated approach for effectively segmenting tubular structure from the chest CT images.

A Key Node Mining Method Based on Acupoint-Disease Network (ADN): A New Perspective for Exploring Acupoint Specificity.

In the process of treating pro-diseases with acupuncture, traditional Chinese medicine (TCM) doctors may fine-tune acupuncture prescriptions according to different prior experiences. Different prescriptions will affect the efficiency and effect of acupuncture treatment, and even excessive acupoint selection may cause psychological pressure on patients. We still lack an effective means to analyze the meridian system and acupoint specificity to clarify the mapping relationship between acupoints and diseases. Given the inability of modern medical technology to provide effective evidence support for meridians and acupoints, we combined acupuncture theory with network science for an interdisciplinary discussion. In this paper, we constructed a weighted undirected acupoint-disease network (ADN) based on clinical acupuncture prescription literature and proposed a high-specificity key node mining method based on ADN. Combined with the principle of acupoint selection in TCM, the proposed method balanced the contribution of local areas to the network based on the distribution characteristics of meridians and selected 30 key acupoints with high influence on the global topology according to the evaluation index of key nodes. Finally, we compared the proposed method with the other six classical node importance evaluation algorithms in terms of resolution, network loss, and accuracy. The comprehensive results show that the marked key acupoint nodes make outstanding contributions to the connectivity, topological structure, and weighted benefits of the network, and the stability and specificity of the algorithm guarantee the reliability of the key acupoint nodes. We consider that these key acupoints with high centrality in ADN can be used as core acupoints to help researchers explore targeted and high-impact acupoint combinations under resource constraints and optimize existing acupuncture prescriptions.

MR-Forest: A Deep Decision Framework for False Positive Reduction in Pulmonary Nodule Detection.

With the development of deep learning methods such as convolutional neural network (CNN), the accuracy of automated pulmonary nodule detection has been greatly improved. However, the high computational and storage costs of the large-scale network have been a potential concern for the future widespread clinical application. In this paper, an alternative Multi-ringed (MR)-Forest framework, against the resource-consuming neural networks (NN)-based architectures, has been proposed for false positive reduction in pulmonary nodule detection, which consists of three steps. First, a novel multi-ringed scanning method is used to extract the order ring facets (ORFs) from the surface voxels of the volumetric nodule models; Second, Mesh-LBP and mapping deformation are employed to estimate the texture and shape features. By sliding and resampling the multi-ringed ORFs, feature volumes with different lengths are generated. Finally, the outputs of multi-level are cascaded to predict the candidate class. On 1034 scans merging the dataset from the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine (AH-LUTCM) and the LUNA16 Challenge dataset, our framework performs enough competitiveness than state-of-the-art in false positive reduction task (CPM score of 0.865). Experimental results demonstrate that MR-Forest is a successful solution to satisfy both resource-consuming and effectiveness for automated pulmonary nodule detection. The proposed MR-forest is a general architecture for 3D target detection, it can be easily extended in many other medical imaging analysis tasks, where the growth trend of the targeting object is approximated as a spheroidal expansion.

[Simvastatin promotes murine osteoclasts apoptosis in vitro through NFATc1 pathway].

OBJECTIVE: To explore the mechanism by which simvastatin (SIM) regulates osteoclast apoptosis. METHODS: Murine macrophage RAW264.7 cells were divided into 5 groups, namely group A (control group), group B (sRANKL+ M-CSF), group C (SIM+sRANKL+M-CSF), group D (VIVIT peptide+sRANKL+ M-CSF), and group E (SIM+VIVIT peptide+sRANKL+M-CSF). WST-1 assay was used to assess the effects of simvastatin on the proliferation activity of the osteoclasts, and flow cytometry was performed to analyze the effects of SIM and VIVIVIT peptide (a NFATc1 pathway inhibitor) on apoptosis of the osteoclasts. The translocation of NFATc1 into the nucleus was investigated using immunofluorescence assay, and Western blotting was employed to assess the effect of SIM on the phosphorylation of NFATc1 in the nucleus. RESULTS: WST-1 assay showed that SIM (1×10-6 mol/L) treatment for 24 and 48 h significantly inhibited the proliferation of the osteoclasts (P=0.039 and 0.022, respectively). Compared with the control group, the SIM-treated osteoclasts exhibited significantly reduced cell percentage in G0/G1 phase (P=0.041) and increased cells in sub-G1 phase (P=0.028) with obvious cell apoptosis. DAPI staining and flow cytometry showed that both SIM and VIVIVIT peptide alone significantly promoted osteoclast apoptosis (P=0.002 and 0.015, respectively), and their combination produced a similar pro-apoptosis effect (P=0.08). Immunofluorescence and Western blotting showed that SIM significantly inhibited the intranuclear translocation of NFATc1 and the phosphorylation of NFATc1 pathway protein (P=0.013). CONCLUSIONS: SIM promotes osteoclast apoptosis through NFATc1 signaling pathway.

LncRNA BANCR promotes tumorigenesis and enhances adriamycin resistance in colorectal cancer.

Colorectal cancer (CRC) is the third most common malignancy in the United States. Chemotherapeutic resistance is a massive obstacle for cancer treatment. The roles and molecular basis of long non-coding RNA BRAF-activated noncoding RNA (BANCR) in CRC progression and adriamycin (ADR) resistance have not been extensively identified. In this study, we found that BANCR and CSE1L expressions were upregulated in CRC tumor tissues. Meanwhile, CSE1L expression was correlated with depth of CRC. BANCR silencing suppressed cell proliferation and invasion capacity, increased apoptotic rate and potentiated cell sensitivity to ADR. CSE1L downregulation triggered a reduction of cell proliferation and invasion ability, and an increase of apoptosis rate and cell sensitivity to ADR. CSE1L overexpression attenuated si-BANCR-mediated anti-proliferation, anti-invasion and pro-apoptosis effects in CRC cells. BANCR acted as a molecular sponge of miR-203 to sequester miR-203 away from CSE1L in CRC cells, resulting in the upregulation of CSE1L expression. CSE1L knockdown inhibited expressions of DNA-repair-related proteins (53BP1 and FEN1) in HCT116 cells. BANCR knockdown also inhibited tumor growth and enhanced ADR sensitivity in CRC mice model. In conclusion, BANCR knockdown suppressed CRC progression and strengthened chemosensitization of CRC cells to ADR possibly by regulating miR-203/CSE1L axis, indicating that BANCR might be a promising target for CRC treatment.

Dynamic compression promotes proliferation and neovascular networks of endothelial progenitor cells in demineralized bone matrix scaffold seed.

Neovascularization is required for bone formation and successful fracture healing. In the process of neovascularization, endothelial progenitor cells (EPCs) play an important role and finish vascular repair through reendothelialization to promote successful fracture healing. In this study, we found that dynamic compression can promote the proliferation and capillary-like tube formation of EPCs in the demineralized bone matrix (DBM) scaffold seed. EPCs isolated from the bone marrow of rats have been cultured in DBM scaffolds before dynamic compression and then seeded in the DBM scaffolds under dynamic conditions. The cells/scaffold constructs were subjected to cyclic compression with 5% strain and at 1 Hz for 4 h/day for 7 consecutive days. By using MTT and real-time PCR, we found that dynamic compression can significantly induce the proliferation of EPCs in three-dimensional culture with an even distribution of cells onto DBM scaffolds. Both in vitro and in vivo, the tube formation assays in the scaffolds showed that the loaded EPCs formed significant tube-like structures. These findings suggest that dynamic compression promoted the vasculogenic activities of EPCs seeded in the scaffolds, which would benefit large bone defect tissue engineering.