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Most basal-like breast cancers (BLBCs) are triple-negative breast cancers (TNBCs), which is associated with high malignancy, high rate of recurrence and distant metastasis, and poor prognosis among all types of breast cancer. However, there are currently no effective therapies for BLBC. Furthermore, chemoresistance limits the therapeutic options for BLBC treatment. In this study, we screen out protein activator of the interferon-induced protein kinase (PACT) as an essential gene in BLBC metastasis. We find that high PACT expression level was associated with poor prognosis among BLBC patients. In vivo and in vitro investigations indicated that PACT could regulate BLBC metastasis by interacting with SUMO-conjugating enzyme Ubc9 to stimulate the SUMOylation and thus consequently the activation of Rac1. BLBC patients receiving chemotherapy presents poorer prognosis with PACT high expression, and PACT disruption sensitizes experimental mammary tumor metastases to chemotherapy, thus providing insights to consider PACT as a potential therapeutic target to overcome acquired chemoresistance in BLBC.

Tumors are one of the most common fatal diseases worldwide and pose a severe threat to human health. Effective tumor prevention and treatment strategies are persistent challenges in the medical community. Angiogenesis plays a critical role in and is the basis for tumor development and metastasis. Circular RNAs (circRNAs) are novel single‑stranded covalently closed RNA molecules that are widely expressed in tumors due to their structural specificity and conservation. circRNAs affect angiogenesis by functioning as microRNA sponges to regulate vascular endothelial growth factor‑related pathways, thereby participating in various stages of tumor growth, invasion and proliferation. The present review summarizes the involvement of circRNAs in the regulation of tumor angiogenesis through competing endogenous RNA mechanisms, with a particular focus on the regulatory role of circRNAs in tumor angiogenesis in various systems. It is considered that circRNAs have great potential for use as tumor diagnostic markers and anti‑angiogenic therapies, and are thus worthy of further research and exploration.

Oxidative stress mediated apoptotic and pyroptotic cell death contributes to intervertebral disc (IVD) degeneration, and platelet-rich plasma (PRP) exerts protective effects to attenuate IVD degeneration. Hence, the present study aimed to validate this issue and uncover the potential underlying mechanisms. The mice and cellular models for IVD degeneration were established by using puncture method and H2O2 exposure, respectively, and we evidenced that NLRP3-mediated cell pyroptosis, apoptosis and inflammatory responses occurred during IVD degeneration progression in vitro and in vivo. Then, the PRP-derived exosomes (PRP-exo) were isolated and purified, and we noticed that both PRP-exo and ROS scavenger (NAC) reversed the detrimental effects of H2O2 treatment on the nucleus pulposus (NP) cells. Further results supported that PRP-exo exerted its protective effects on H2O2 treated NP cells by modulating the Keap1-Nrf2 pathway. Mechanistically, PRP-exo downregulated Keap1, resulting in the release of Nrf2 from the Keap1-Nrf2 complex, which further translocated from cytoplasm to nucleus to achieve its anti-oxidant biological functions, and H2O2 treated NP cells with Nrf2-deficiency did not respond to PRP-exo treatment. In addition, miR-141-3p was enriched in PRP-exo, and miR-141-3p targeted the 3' untranslated region (3'UTR) of Keap1 mRNA for its degradation, leading to Nrf2 translocation. Furthermore, overexpression of miR-141-3p ameliorated the cytotoxic effects of H2O2 on NP cells, which were abrogated by upregulating Keap1 and silencing Nrf2. Taken together, we concluded that PRP secreted exosomal miR-141-3p to activate the Keap1-Nrf2 pathway, which helped to slow down IVD degeneration.

Super activated platelet lysate (sPL) is a derivative of platelet-rich plasma (PRP) that contains high levels of several growth factors. In this study, we synthesized a temperature-sensitive hydrogel that contained temperature-sensitive Poly(DL-lactide-glycolide-glycolide acid) (PLGA), SrCl2, and HA, and loaded it with different concentrations of sPL. The hydrogel showed satisfactory encapsulation efficiency and release of the growth factors in a sustained manner, indicating its suitability as a drug carrier. The sPL-loaded hydrogel was inserted into the necrotic femoral head of a rat model and core decompression was applied and resulted in significantly accelerated bone repair and regeneration. Therefore, encapsulation of sPL in a hydrogel scaffolding may be an effective strategy for treating femoral head necrosis.

ObjectiveTo evaluate the therapeutic effect of platelet lysate (PL)/vancomycin/nano-hydroxyapatite sustained-release material on treating staphylococcus aureus-induced infectious bone defects.Methods40 New Zealand white rabbits were inoculated with staphylococcus aureus to construct the chronic tibial infectious bone defect model. After incision, debridement and washing, control group 1 was not given any filling, control group 2 was filled with PL/nano-hydroxyapatite sustained release material, control group 3 was filled with vancomycin/ nano-hydroxyapatite sustained release material, and the treatment group was filled with PL/vancomycin/nano-hydroxyapatite sustained-release material. Afterwards, the drug release profiles were determined in vitro and in vivo. Then, X-rays and bone specimens were used to evaluate the efficacy of the treatments.ResultsTGF-β and PDGF were effectively released for 28 days in vitro. In addition, results of the inhibition zone experiment of the composite material proved that vancomycin had favorable antibacterial activity, which effectively suppressed bacteria for as long as 43 days, thus achieving the sustained-release antibacterial effect. The drug release profiles in vitro also demonstrated that the vancomycin concentration within the lesion region was the highest in composite material, and the infection in experimental rabbits was markedly alleviated. The original backbone deformity regained the normal shape, the normal bone marrow structure began to recover 6 weeks later, and the nano-hydroxyapatite transformed into the trabecula structure. By contrast, the inflammation in the control group still existed, with no obvious new bone formation.ConclusionThe PL/vancomycin/nano-hydroxyapatite sustained-release material effectively treats chronic infectious bone defects.

Background: Kashin-Beck disease (KBD) is currently an endemic form of osteoarthritis. In this study, we explored novel KBD diagnostic biomarkers.Methods: The GSE59446 dataset was used to conduct Weighted Gene Co-expression Network Analysis (WGCNA) and differentially expressed genes (DEGs) analysis with peripheral blood samples of 100 healthy individuals and 100 KBD patients. As part of the gene ontology pathway enrichment analysis, genes related to SONFH and DEGs were selected from the extraction module. Then, central DEGs were selected for LASSO analysis, and, based on SVM-RFE and DEG results, overlapping genes were identified as key KBD genes. Next, we analyzed the correlations between the selected genes and age, gender, and other factors to eliminate their influences on gene expression. Finally, we evaluated the diagnostic value of key KBD genes using case information collected by us.Results: Seven gene co-expression modules were created using WGCNA. The turquoise module was identified as a KBD key module since it showed the highest correlation to KBD. The functional enrichment analysis revealed that the genes associated with this key module were mainly involved in mitochondrial reactions, protein heterooligomerization, and negatively regulating cysteine-type endopeptidase-dependent apoptotic processes. Additionally, 12 key genes were identified using the LASSO analysis, 5 major genes using SVM-RFE analysis, and 36 DEGs were screened through the "limma" R package. The GLRX5 gene - pivotal in DEGs, LASSO, and SVM-RFE - was further aggregated as the key KBD gene. Correlation analyses confirmed the GLRX5 diagnostic value for KBD and that it was not related to age, gender, and other factors. Finally, data from our patients demonstrated that GLRX5 can be a KBD diagnostic biomarker.Conclusions: We demonstrated that the target gene GLRX5 can be a KBD non-invasive diagnosis biomarker.

Background:The high incidence and mortality of lung cancer has been a widespread concern in the world, among which non-small cell lung cancer (NSCLC)accounts for more than 85% of lung cancer, and its 5-year prognosis is pretty poor. Accumulating studies have shown that traditional Chinese medicine (TCM) and its active ingredients have shown good anti-tumor activity. Based on the results of our previous high-throughput screening, it was the first time to find that Thevebioside (THB, an active ingredient from TCM) is sensitive to NSCLC cells, and then we will still the first to reveal the underling mechanism of its tumor suppressive activity.Methods: In this study, MTT and colony formation assay were used to detect cell growth. Annexin V/ propidium iodide, Hoechst 33342 were applied to test the process of cellular apoptosis or cell cycle arrest which determined by flow cytometry in NSCLC (A549 and H460) cells. qRT-PCR and Western blot was utilized to value the level of expression of SRC&(SRC1-3) and apoptosis related proteins. The ubiquitination degradation of SRC3 was through proteasome dependent pathway were examined by the usage of CHX and MG132 and the co-immunoprecipitation assay. The anticancer activity of THB, Neritaloside，Odoroside H and digoxin in vivo were monitored in nude mice by subcutaneous inoculation or intrapulmonary injection of A549 cells and then administrated these agents (2 mg/ kg or 4mg/kg) by intraperitoneal injection three times a week for 30 days. Tumor growth was monitored by a luciferase living imaging system. Body weight of mice and tumor volume were measured weekly. The survival carve was recorded. Pathological histology of liver, kidney and heart were detected by H&E staining, and its functions were tested by ELISA. Results：THB effectively inhibits the outgrowth of NSCLC cells (A549 and H460) by inducing cellular apoptosis. SRC-3 was significantly down-regulated after THB treatment dependent on ubiquitin-proteasome-mediated degradation, which subsequently inhibited the AKT signaling pathway to promte apoptosis. Animal study supported in vitro results, showing THB had a good suppressive effect on tumor growth. Besides, it also indicated that THB without overt cytotoxic effect on liver, kidney and heart.Conclusion：THB , as an active extract from TCM，exerted inhibitory effect on tumor growth of NSCLC by down-regulating the expression of SCR-3 rely on the ubiquitin-proteasome degradation pathway, thereby inhibiting the AKT signaling to induce cellular apoptosis , with minimal toxicity.

Background Platelet lysate (PL) had a remarkable therapeutic effect on bone repair related diseases, such as delayed fracture healing, femoral head necrosis and meniscal tear. In this study, we investigated the effect of PL on patients with nonunion, cartilage repair and osteonecrosis, and to evaluate the effect of PL on nonunion cells proliferation and the effect of PL on OPG/RANKL signaling pathway in nonunion cell of male rats. To reveal the molecular mechanism of PL for bone healing. Methods We used different concentrations of PL to treat nonunion cells, then detected cell proliferation and protein expression levels of osteoprotegerin (OPG), RANKL, osteopontin (OPN), osteocalcin (OCN) and alkaline phosphatase (ALP). Results The proliferation rate of nonunion cells treated by 5% PL, was significantly higher than that of the control group (P0.05). the results of western blot analysis and immunofluorescence analysis showed that PL improved the expression of OPG, OPN, OCN and ALP proteins in nonunion cells, but PL had no effect on the expression of nuclear factor-κB ligand (RANKL) protein. Conclusions We found that PL had a remarkable therapeutic effect on bone repair related diseases; 5% PL significantly improved the proliferation rate of the nonunion cells; 10% PL had a significantly positive effect on improving the expression levels of osteogenic related genes.

Aims Chondrocyte degeneration is the main cause of osteoarthritis (OA) and increased evidence suggests that miRNAs could have vital roles in the pathology of various cartilage illnesses. miR-1236 has been found to contribute to inflammation in diseases such as pneumonia. However, the exact role of miR-1236 in OA is poorly understood. Materials and methods H&E staining and saffron fixation experiments were employed to determine OA tissues. qRT-PCR and immunohistochemistry were used to detect the expression levels of miR-1236 and PIK3R3. Western blot was performed to detect the expression levels of proteins. Luciferase reporter assays were utilized to investigate the interaction between miR-1236 and PIK3R3. Cell counting assays and AO/EB were used to quantify cell growth and apoptosis. Key findings miR-1236 was up-regulated in OA knee cartilage compared to normal cartilage. Up-regulated expression of miR-1236 suppressed cell proliferation as well as induced apoptosis in chondrocytes. Bioinformatics identified PIK3R3 as a target of miR-1236. Co-transfection with miR-1236 and PIK3R3 could reverse cell apoptosis induced by the miR-1236 mimic. Significance These data enhance our understanding on the role of miR-1236 in OA and identifies miR-1236 as a potential biomarker or possible treatment target within OA.

When dealing with multivariate survival data, featuring the association structure is a key difference from the univariate survival analysis. In this paper, we explore to use the composite likelihood framework to handle multivariate survival data, where only the lower dimensional survival distributions need to be specified. The development allows us to use available modeling schemes for bivariate survival data to characterize association structures of correlated survival times. The inference procedure is based on the pseudolikelihood which is the product of the lower dimensional bivariate distributions. The proposed estimation procedure is assessed through simulation studies. As a genuine application, we apply the composite likelihood inference procedure to analyze the data from the polybrominated diphenyl ethers (PBDEs) study, where four types of PBDE congeners are available. The associations among the four PBDE congeners, and the relationships between the covariates and the PBDE congeners are of interest. The result shows that there is strong association among the concentrations of the four PBDE congeners, and statistically significant predictors on the concentrations of the four PBDE congeners are identified.

Objective The precise mechanism of nucleus pulposus proliferation in the degeneration of the intervertebral disk pathogenesis remains to be implicated. MicroRNAs (MiRNAs) are a class of 18−22 nucleotides, which are small, noncoding RNAs that inhibit protein translation by binding to the 3′-UTR of target gene. Recent studies have shown that miRNAs play a crucial role in various cell biologies such as cell proliferation, invasion, migration, and cell cycle. However, the role of miR-184 in nucleus pulposus proliferation is still unknown. Method qRT-PCR was performed to measure the expression of miR-184. CCK-8 assay, qRT-PCR, and Western blot were used to measure the functional role of miR-184 in nucleus pulposus (NP) cells. Western blot and Luciferase assays were done to find the miR-184 target gene. Result We demonstrated that expression of miR-184 was upregulated in degenerative NP tissues compared with that in the control NP tissues, and the expression of miR-184 was positively correlated with disk degeneration grade. We identified Growth Arrest Specific Gene 1 ( GAS1 ) as a direct target gene of miR-184 in NP cells, and ectopic expression of miR-184 promoted NP cells proliferation. In addition, we found that GAS1 expression was downregulated in degenerative NP tissues compared with that in the control NP tissues and the GAS1 expression was inversely correlated with the grade of disk degeneration. Moreover, we demonstrated that miR-184 overexpression could induce AKT phosphorylation and ectopic expression of GAS1 decreased the miR-184 overexpressing NP cells proliferation. Conclusion These results demonstrated that miR-184 and the GAS1 /Akt pathway may be a potential therapeutic target for intervertebral disc degeneration.

We investigate claims and assumptions made in several recent papers about fault localization (FL) techniques. Most of these claims have to do with evaluating FL accuracy. Our investigation centers on a new subject program having properties useful for FL experiments. We find that Tarantula (Jones et al.) works well on the program, and we show weak support for the assertion that coverage-based test suites help Tarantula to localize faults. Baudry et al. used automatically-generated mutants to evaluate the accuracy of an FL technique that generates many distinct scores for program locations. We find no evidence to suggest that the use of mutants for this purpose is invalid. However, we find evidence that the standard method for evaluating FL accuracy is unfairly biased toward techniques that generate many distinct scores, and we propose a fairer method of accuracy evaluation. Finally, Denmat et al. suggest that data mining techniques may apply to FL. We investigate this suggestion with the data mining tool Weka, using standard techniques for evaluating the accuracy of data mining classifiers. We find that standard classifiers suffer from the class imbalance problem. However, we find that adding cost information improves accuracy.