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Background Glioblastoma multiforme, the most aggressive and malignant primary brain tumor, is characterized by rapid growth and extensive infiltration to neighboring normal brain parenchyma. Our previous studies delineated a crosstalk between PI3K/Akt and JNK signaling pathways, and a moderate anti-glioblastoma synergism caused by the combined inhibition of PI3K p110β (PI3Kβ) isoform and JNK. However, this combination strategy is not potent enough. MLK3, an upstream regulator of ERK and JNK, may replace JNK to exert stronger synergism with PI3Kβ. Methods To develop a new combination strategy with stronger synergism, the expression pattern and roles of MLK3 in glioblastoma patient’s specimens and cell lines were firstly investigated. Then glioblastoma cells and xenografts in nude mice were treated with the PI3Kβ inhibitor AZD6482 and the MLK3 inhibitor URMC-099 alone or in combination to evaluate their combination effects on tumor cell growth and motility. The combination effects on cytoskeletal structures such as lamellipodia and focal adhesions were also evaluated. Results MLK3 protein was overexpressed in both newly diagnosed and relapsing glioblastoma patients’ specimens. Silencing of MLK3 using siRNA duplexes significantly suppressed migration and invasion, but promoted attachment of glioblastoma cells. Combined inhibition of PI3Kβ and MLK3 exhibited synergistic inhibitory effects on glioblastoma cell proliferation, migration and invasion, as well as the formation of lamellipodia and focal adhesions. Furthermore, combination of AZD6482 and URMC-099 effectively decreased glioblastoma xenograft growth in nude mice. Glioblastoma cells treated with this drug combination showed reduced phosphorylation of Akt and ERK, and decreased protein expression of ROCK2 and Zyxin. Conclusion Taken together, combination of AZD6482 and URMC-099 showed strong synergistic anti-tumor effects on glioblastoma in vitro and in vivo. Our findings suggest that combined inhibition of PI3Kβ and MLK3 may serve as an attractive therapeutic approach for glioblastoma multiforme.

Additional file 1: Table S1. WHO classification and IDH mutation status of glioma cell lines applied in our study. Table S2. Clinical information of the LGG and GBM cohort included for our OS analysis from TCGA database.

Additional file 2: Figure S1. a–c The role of Skp2 expression in the OS of patients with IDH1wt GBM, IDHwt LGG and even IDHmut LGG was analyzed. d, e The role of Skp2 expression in the OS of IDHmut LGG patients with or without 1p19q co-deletion was analyzed. Figure S2. Cell cycle cascade rate were detected in three cell lines, U87, U138 and LNZ308, upon LV, TMZ or combined treatments. With regard to the strong effect of TMZ in glioma cells, we chose 10 nM as the combination concentration of lovastatin in our study. Lovastatin caused a little bit of G2/M arrest in U87 and LNZ308, but not in U138 cell. The G1 phase cell numbers were enhanced in U138 and LNZ308 cells upon lovastatin treatment. Lovastatin did not further induce G2/M arrest when combined with TMZ in U87 and U138. However, in LNZ308 cells, lovastatin promoted G2/M arrest upon TMZ treatments dramatically. (*p

Heavy metal pollution is a global ecological safety issue, especially in crops, where it directly threatens regional ecological security and human health. In this study, the back-propagation (BP) neural network optimized by the genetic algorithm (GA) was used to predict the concentration of cadmium (Cd) in rice grain based on influencing factors. As an intelligent information processing system, the GA-BP neural network could learn the laws of Cd movement in the soil-crop system through its own training and use the soil properties to predict the concentration of Cd in grain with high accuracy. The total soil Cd concentration, clay content, Ni concentration, cation exchange capacity (CEC), organic matter (OM), and pH have important impacts and interactions on Cd concentration in rice grain were selected as input factors of the prediction model based on Pearson's correlation analysis and GeoDetector. By using GA to optimize the initial weight, the prediction accuracy of the GA-BP neural network model was optimal compared with the BP neural network model and multiple regression analysis. Based on the Cd concentration predicted in grain by the model, human exposure and health risk can be assessed quickly, enabling measures to be taken in time to reduce the transfer of Cd from soil to the food chain.

Background: Glioblastoma multiforme (GBM) is the most aggressive and malignant primary brain tumor characterized by rapid growth and extensive infiltration to neighboring normal brain parenchyma, which contribute to tumor recurrence and poor prognosis. Myricetin is a natural flavonoid with potent anti-oxidant, anti-inflammatory and anti-cancer activities, which may serve as a potential and harmless agent for GBM treatment. Methods: To investigate the anti-glioblastoma effects of myricetin, GBM cells were treated with myricetin alone or in combination with temozolomide. Its effects on GBM cell motility and cytoskeletal structures including lamellipodia, focal adhesions and membrane ruffles were also evaluated. Results: We showed that myricetin alone inhibited glioblastoma U-87 MG cell proliferation, migration and invasion, whereas combination of myricetin and temozolomide did not exhibit any synergistic effect. The inhibitory effect on GBM cell proliferation is independent of PTEN status. Moreover, myricetin showed less cytotoxicity to normal astrocytes than GBM cells. Formation of lamellipodia, focal adhesions, membrane ruffles and vasculogenic mimicry were blocked by myricetin, and phosphorylation of ROCK2, paxillin and cortactin was suppressed. In addition, myricetin could inhibit PI3K/Akt and JNK signaling, and bind to a series of kinases and scaffold proteins including PI3K catalytic isoforms (p110α, p110β and p110δ), PDK1, JNK, c-Jun, ROCK2, paxillin, vinculin and VEcadherin. Conclusions: In conclusion, myricetin is a multi-targeted drug that has potent anti-migratory and antiinvasive effects on GBM cells, and suppresses formation of lamellipodia and focal adhesions, suggesting that it may serve as an alternative option for GBM treatment.

Osteosarcoma (OS) is the most common primary aggressive and malignant bone tumor. Newly diagnostic OS patients benefit from the standard therapy including surgical resection plus radiotherapy and neoadjuvant chemotherapy (MAP chemotherapy: high-dose methotrexate, doxorubicin and cisplatin). However, tumor recurrence and metastasis give rise to a sharp decline of the 5-year overall survival rate in OS patients. Little improvement has been made for decades, urging the development of more effective therapeutic approaches. ErbB receptor family including EGFR, HER2, HER3 and HER4, being important to the activation of PI3K/Akt and MAPK signaling pathways, are potential targets for OS treatment. Genetic aberrations (amplification, overexpression, mutation and altered splicing) of ErbB are essential to the growth, apoptosis, motility and metastasis in a variety of cancers. Overexpression of ErbB family is associated with the poor prognosis of cancer patients. A number of monoclonal antibodies or inhibitors specific for ErbB family have entered clinical trials in a range of solid tumors including breast carcinoma, lung carcinoma and sarcoma. Here, we summarized the roles and expression of ErbB family in OS and the current development of ErbB-targeted therapeutic strategies including chemotherapies and immunotherapies for OS treatment.

Glioma is the most frequent central nervous system tumor in adults. The overall survival of glioma patients is disappointing, mostly due to the poor prognosis of glioblastoma (Grade IV glioma). Isocitrate dehydrogenase (IDH) is a key factor in metabolism and catalyzes the oxidative decarboxylation of isocitrate. Mutations in IDH genes are observed in over 70% of low-grade gliomas and some cases of glioblastoma. As the most frequent mutation, IDH1(R132H) has been served as a predictive marker of glioma patients. The recently developed droplet digital PCR (ddPCR) technique generates a large amount of nanoliter-sized droplets, each of which carries out a PCR reaction on one template. Therefore, ddPCR provides high precision and absolute quantification of the nucleic acid target, with wide applications for both research and clinical diagnosis. In the current study, we collected 62 glioma tissue samples (Grade II to IV) and detected IDH1 mutations by Sanger direct sequencing, ddPCR, and quantitative real-time PCR (qRT-PCR). With the results from Sanger direct sequencing as the standard, the characteristics of ddPCR were compared with qRT-PCR. The data indicated that ddPCR was much more sensitive and much easier to interpret than qRT-PCR. Thus, we demonstrated that ddPCR is a reliable and sensitive method for screening the IDH mutation. Therefore, ddPCR is able to applied clinically in predicting patient prognosis and selecting effective therapeutic strategies. Our data also supported that the prognosis of Grade II and III glioma was better in patients with an IDH mutation than in those without mutation.

Glioblastoma multiforme (GBM) is thought to result from an imbalance between glucose metabolism and tumor growth. The Myc oncogene and lethal-7a microRNA (let-7a miRNA) have been suggested to cooperatively regulate multiple downstream targets leading to changes in chromosome stability, gene mutations, and/or modulation of tumor growth. Here, we review the roles of Myc and let-7a in glucose metabolism and tumor growth and addresses their future potential as prognostic markers and therapeutic tools in GBM. We focus on the functions of Myc and let-7a in glucose uptake, tumor survival, proliferation, and mobility of glioma cells. In addition, we discuss how regulation of different pathways by Myc or let-7a may be useful for future GBM therapies. A large body of evidence suggests that targeting Myc and let-7a may provide a selective mechanism for the deregulation of glucose metabolic pathways in glioma cells. Indeed, Myc and let-7a are aberrantly expressed in GBM and have been linked to the regulation of cell growth and glucose metabolism in GBM. This article is part of a Special Issue entitled "Targeting alternative glucose metabolism and regulate pathways in GBM cells for future glioblastoma therapies".

The phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway and c-Jun N-terminal kinase (JNK) pathway are responsible for regulating a variety of cellular processes including cell growth, migration, invasion and apoptosis. These two pathways are essential to the development and progression of tumors. The dual roles of JNK signaling in apoptosis and tumor development determine the different interactions between the PI3K/Akt and JNK pathways. Activation of PI3K/Akt signaling can inhibit stress- and cytokine-induced JNK activation through Akt antagonizing and the formation of the JIP1-JNK module, as well as the activities of upstream kinases ASK1, MKK4/7 and MLK. On the other hand, hyperactivation of Akt and JNK is also found in cancers that harbor EGFR overexpression or loss of PTEN. Understanding the activation mechanism of PI3K/Akt and JNK pathways, as well as the interplays between these two pathways in cancer may contribute to the identification of novel therapeutic targets. In the present report, we summarized the current understanding of the PI3K/Akt and JNK signaling networks, as well as their biological roles in cancers. In addition, the interactions and regulatory network between PI3K/Akt and JNK pathways in cancer were discussed.

Many drug candidates with high therapeutic efficacy have low water solubility, which limits the administration and transport across physiological barriers, for example, the tumor tissue barrier. Therefore, strategies are needed to permeabilize the physiological barriers safely so that hydrophobic drugs may be delivered efficiently.This review focuses on prospects for therapeutic application of lipid-based drug delivery carriers that increase hydrophobic drugs to improve their solubility, bioavailability, drug release, targeting and absorption. Moreover, novel techniques to prepare for lipid-based drug delivery to extend pharmaceuticals with poor bioavailability such as surface modifications of lipid-based drug delivery are presented. Industrial developments of several drug candidates employing these strategies are discussed, as well as applications and clinical trials.Overall, hydrophobic drugs can be encapsulated in the lipid-based drug delivery systems, represent a relatively safe and promising strategy to extend drug retention, lengthen the lifetime in the circulation, and allow active targeting to specific tissues and controllable drug release in the desirable sites. However, there are still noticeable gaps that need to be filled before the theoretical advantage of these formulations may truly be realized such as investigation on the use of lipid-based drug delivery for administration routes. This research may provide further interest within the area of lipid-based systems, both in industry and in the clinic.

Characteristics of isoform-selective PI3K inhibitors. Supplementary Table 2. Characteristics of JNK inhibitor SP600125. Supplementary Table 3. Combination index (CI) for cell viability in GBM cell line U-87 MG in the combination of PIK-75 and SP600125 (n = 3). Supplementary Fig. 1: Half-maximal inhibitory concentration (IC50) values of PIK-75, TGX-221, CAL-101 and SP600125 on glioblastoma cell line U87-MG. Supplementary Fig. 2. Effects of PIK-75, TGX-221, CAL-101 and SP600125 on cell viability of glioblastoma cell line U-373 MG. Supplementary Fig. 3. Combination effects of isoform-selective PI3K inhibitors and SP600125 on proliferation of glioblastoma cells and normal human astrocytes. Supplementary Fig. 4. Combination effects of isoform-selective PI3K inhibitors and SP600125 on migration of U-373 MG glioblastoma cells. Supplementary Fig. 5. Combination effects of isoform-selective PI3K inhibitors and SP600125 on invasion of U-373 MG glioblastoma cells. Supplementary Fig. 6. Combination effects of isoform-selective PI3K inhibitors and SP600125 on glioblastoma U-87 MG cell apoptosis. (DOC 3768 kb)

The antibacterial and biological properties of silver-loated coralline hydroxyapatite (Ag-CHA) as a new antibacterial implant material were investigated in this study. Compared to other antibiotic and chemical bactericidal agents, Ag+ does not bring bacterial resistance to drugs and has less toxicity. The porous CHA was formed by hydrothermal exchange, then Ag+ was loated onto CHA through ion exchange and adsorption. The microstructure and composition of Ag-CHA were characterized by scanning electron microscopy (SEM), Rutherford backscattering spectrometry (RBS), and energy dispersive spectrometry (EDS). Antibacterial activity of Ag-CHA on the clinical strains of Escherichia coli (E. Coli) and Staphylococcus aureus (S. Aureus) was evaluated by the flat plate diffusion method. The antibacterial activity of Ag-CHA was found to be correlated with the concentration of Ag+ in a dose-dependent manner, which indicated that the optimal antibacterial and biocompatible effects of Ag-CHA could be obtained with Ag+ concentrations from 5×10−5 to 1×10−4 mol/L.

Six groups of grass carp (average weight 266.9 ± 0.6 g) were fed diets containing 197, 385, 770, 1082, 1436 and 1795 mg choline/kg, for 8 weeks. Fish growth, and muscle nutrient (protein, fat and amino acid) content of young grass carp were significantly improved by appropriate dietary choline. Furthermore, muscle hydroxyproline concentration, lactate content and shear force were improved by optimum dietary choline supplementation. However, the muscle pH value, cooking loss and cathepsins activities showed an opposite trend. Additionally, optimum dietary choline supplementation attenuated muscle oxidative damage in grass carp. The muscle antioxidant enzyme (catalase and glutathione reductase did not change) activities and glutathione content were enhanced by optimum dietary choline supplementation. Muscle cooking loss was negatively correlated with antioxidant enzyme activities and glutathione content. At the gene level, these antioxidant enzymes, as well as the targets of rapamycin, casein kinase 2 and NF-E2-related factor 2 transcripts in fish muscle were always up-regulated by suitable choline. However, suitable choline significantly decreased Kelch-like ECH-associated protein 1 a (Keap1a) and Kelch-like ECH-associated protein 1 b (Keap1b) mRNA levels in muscle. In conclusion, suitable dietary choline enhanced fish flesh quality, and the decreased cooking loss was due to the elevated antioxidant status that may be regulated by Nrf2 signaling.

Gliomas are a common adult central nervous system tumor, and glioblastoma (GBM), which has a poor prognosis, is the most lethal of all gliomas. The overall survival of GBM patients is only 12-14 months after diagnosis. With progress in the precision of personal medication, therapeutic options for various tumors have become gradually dependent on the molecular profiles of patients. GBM is one of the tumors in which treatment response relies largely on the molecular characteristics of the tumor. Therefore, awareness of the genetic background of each patient will help decision-making regarding the best treatment strategy to use. In this review, a novel molecular classification of gliomas based on recent findings of their genetic characteristics is introduced. Representative molecular markers, such as IDH1 mutation, 1p19q co-deletion, MGMT promoter methylation and EGFRvIII amplification, are described. Furthermore, the development of non-coding RNAs and omics studies of GBM are briefly discussed. Finally, a novel concept for non-invasive detection that could facilitate both diagnosis and treatment monitoring is presented. There is no doubt that the use of molecular profiling by biomarkers will indeed improve the overall survival and quality of life of GBM patients.

Since 1984, Beijing’s rapid economic growth and urbanization have been heading into resource and environmental problems, such as cropland decrease, environmental deterioration, and traffic jams. Therefore, more space for glades and trees is needed to improve the urban environment. However, the central government’s decision to maintain 227,000 ha of cropland for national food security created a dilemma for Beijing: the need to choose between keeping the cropland or planting more trees and enlarging the glades. Actually, people can use the croplands to serve both purposes—food production and eco-environmental service—because croplands are multifunctional anyway. Besides food production, croplands also have ecological, landscape, cultural, and tourism roles, especially in megacities. This chapter analyzed land use and crop structure based on the land-use survey and agricultural statistic data while discussing the existing state of agriculture, including economic profit, labor cost, water resource depletion, and environmental impact. This study then proposed a four-ring agricultural pattern that radiates outward from the city center and exploits the multifunctional nature of croplands: an inner-city ring providing urban beautification with flowers and grass; a city-skirt ring with fruit trees, grain crops, and grass, from which intensive agriculture should be prohibited; a suburban plain ring for efficient, intensive agriculture; and lastly, the mountainous regions, for ecological conservation and unique agricultural products. This way, cultivation is combined with landscaping and ecological protection to give Beijing a more comfortable and beautiful living environment. The proposed spatial planning of agriculture is different from the traditional agricultural ring pattern identified by Thunen in 1826, but it inherits and develops the thought illustrated in Garden Cities of Tomorrow, a book written by Howard in 1945.

Gang Wang,1,2,* Jun Jie Wang,1,2,* Tony SS To,3 Hua Fu Zhao,3 Jing Wang3 1Department of Pharmaceutics, Shanghai Eighth People’s Hospital, Shanghai, People’s Republic of China; 2College of Pharmacy, Hubei University of Medicine, Shiyan, Hubei Province, People’s Republic of China; 3Department of Health Technology and Informatics, The Hong Kong Polytechnic University, Hong Kong SAR, People’s Republic of China *These authors contributed equally tothis work Abstract: Flavonoids, the major polyphenol components in Cotinus coggygria (CC), have been found to show an anticancer effect in our previous study; however, the exact mechanisms of inducing human glioblastoma (GBM) cell death remain to be resolved. In this study, a novel polyvinylpyrrolidone K-30/sodium dodecyl sulfate and polyethyleneglycol-coated liposome loaded with CC flavonoids (CCFs) was developed to enhance solubility and the antibrain tumor effect, and the molecular mechanism regarding how CCF nanoliposomes (CCF-NLs) induce apoptotic cell death in vitro was investigated. DBTRG-05MG GBM cell lines treated with CCF-NLs showed potential antiproliferative effects. Regarding the underlying mechanisms of inducing apoptosis in DBTRG-05MG GBM cells, CCF-NLs were shown to downregulate the expression of antiapoptotic B-cell lymphoma/leukemia 2 (Bcl-2), an apoptosis-related protein family member, but the expression of proapoptotic Bcl-2-associated X protein was enhanced compared with that in controls. CCF-NLs also inhibited the activity of caspase-3 and -9, which is the initiator caspase of the extrinsic and intrinsic apoptotic pathways. Blockade of caspase activation consistently induced apoptosis and inhibited growth in CCF-NL-treated DBTRG-05MG cells. This study further investigated the role of the Akt pathway in the apoptotic cell death by CCF-NLs, showing that CCF-NLs deactivated Akt. Specifically, CCF-NLs downregulated the expression of p-Akt and SIRT1 as well as the level of phosphorylated p53. Together, these results indicated SIRT1/p53-mediated cell death was induced by CCF-NLs, but not by extracellular signal-regulated kinase, in DBTRG-05MG cells. Overall, this study suggested caspase-dependent activation of both the intrinsic and extrinsic signaling pathways, probably through blockade of the SIRT1/p53-mediated mitochondrial and Akt pathways to exert the proapoptotic effect of CCF-NLs in DBTRG-05MG GBM cells. Keywords: Cotinus coggygria flavonoid nanoliposomes, cell death, SIRT1, mitochondrial, PI3K/Akt pathway

Glioblastoma multiform (GBM) is the most aggressive malignant primary tumor in the central nervous system. Due to the hyperactivation of phosphatidylinositol 3-kinase (PI3K) and c-Jun N-terminal kinase (JNK) pathways in GBM, their respective inhibitors become promising and actively researched therapeutic agents. Our previous study showed that the PI3K catalytic isoform p110δ is highly expressed in GBM cell lines, indicating that p110δ may be a potential target for therapy. However, a single drug may not be sufficient to affect all pathogenic processes such as proliferation, migration and invasion in GBM cells. Therefore, we aimed to evaluate the combination effect of PI3Kδ inhibitor CAL-101 and JNK inhibitor SP600125 on glioma cells. The half maximal inhibitory concentration (IC50) of CAL-101 and SP600125 was first determined by the MTT method in GBM cell line U-87 MG. The combination effect of these 2 inhibitors on cell proliferation was analyzed using Compusyn software and the combination index (CI) was calculated. CI 1.1 indicates an antagonistic effect, and CI between 0.9 and 1.1 indicates an additive effect. Cell migration and invasion capacity of U-87 MG cells were then evaluated under combined treatment with CAL-101 and SP600125. Our results showed that the effect of CAL-101 or SP600125 on cell proliferation was dose- and time-dependent. The IC50 of CAL-101 and SP600125 were 10.15 μM and 19.60 μM respectively. Treatment with SP600125 alone significantly suppressed U-87 MG cell proliferation, and combining 10 μM of CAL-101 and 20 μM of SP600125 showed a moderate synergistic effect. The cytotoxicity of CAL-101 or SP600125 alone, as well as combining them together was low on normal astrocytes. Western blot analysis of the PI3K/Akt and JNK pathways indicated that CAL-101 inhibited Akt phosphorylation at Ser473 and Thr308, whereas SP600125 suppressed JNK activity and Akt phosphorylation at Thr308. Combination of CAL-101 and SP600125 further inhibited Akt phosphorylation at Thr308, but showed no effect on the inhibition of Akt phosphorylation at Ser473 and c-Jun phosphorylation. CAL-101 significantly reduced migration but had no effect on invasion of U-87 MG cells, whereas SP600125 significantly suppressed both their migration and invasion abilities. Combining CAL-101 and SP600125 augmented the inhibitory effect on migration but showed no enhancement on invasion. In conclusion, combination treatment with CAL-101 and SP600125 showed a moderate synergistic effect on GBM cell proliferation, and enhanced the inhibition of cell migration. Our results suggest that GBM patients may benefit from combination therapy targeting both PI3Kδ and JNK. Citation Format: Hua-Fu ZHAO, Jing WANG, Shing-Shun Tony TO. Synergistic effect of PI3Kδ inhibitor CAL-101 and JNK inhibitor SP600125 on glioblastoma cell proliferation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2660. doi:10.1158/1538-7445.AM2015-2660

Background Glioblastoma multiforme is the most aggressive malignant primary brain tumor, characterized by rapid growth and extensive infiltration to neighboring normal brain parenchyma. Both PI3K/Akt and JNK pathways are essential to glioblastoma cell survival, migration and invasion. Due to their hyperactivation in glioblastoma cells, PI3K and JNK are promising targets for glioblastoma treatment. Methods To investigate the combination effects of class IA PI3K catalytic isoforms (p110α, p110β and p110δ) and JNK inhibition on tumor cell growth and motility, glioblastoma cells and xenografts in nude mice were treated with isoform-selective PI3K inhibitors in combination with JNK inhibitor. Results We showed that combined inhibition of these PI3K isoforms and JNK exerted divergent effects on the proliferation, migration and invasion of glioblastoma cells in vitro. Pharmacological inhibition of p110β or p110δ, but not p110α, displayed synergistic inhibitory effect with JNK inhibition on glioblastoma cell proliferation and migration through decreasing phosphorylation of Akt, FAK and zyxin, leading to blockade of lamellipodia and membrane ruffles formation. No synergistic effect on invasion was observed in all the combination treatment. In vivo, combination of p110β and JNK inhibitors significantly reduced xenograft tumor growth compared with single inhibitor alone. Conclusion Concurrent inhibition of p110β and JNK exhibited synergistic effects on suppressing glioblastoma cell proliferation and migration in vitro and xenograft tumor growth in vivo. Our data suggest that combined inhibition of PI3K p110β isoform and JNK may serve as a potent and promising therapeutic approach for glioblastoma multiforme. Electronic supplementary material The online version of this article (doi:10.1186/s13046-016-0356-5) contains supplementary material, which is available to authorized users.