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Cambogin is a polycyclic polyprenylated acylphoroglucinol (PPAP) from the Garcinia genus, which has been used traditionally for cancer treatment across Southeastern Asia. In this study, we found that cambogin inhibited breast cancer cell proliferation and induced cell apoptosis in vitro. Cambogin induced the activation of the caspase-independent mitochondrial apoptotic pathway, as indicated by an increase in the ratio of Bax/Bcl-2 and the nuclear translocation of apoptosis inducing factor (AIF). Two-dimensional gel electrophoresis and mass spectrometry revealed that the expression of proteins involving in the radical oxygen species (ROS) pathway was among the most affected upon cambogin treatment. Cambogin enhanced cellular ROS production, and induced the activation of the ASK1–MKK4/MKK7–JNK/SAPK signaling pathway. Pretreatment with ROS scavenger N-acetylcysteine (NAC), an antioxidant, or the JNK inhibitor SP600125 was able to restore cell viability in the presence of cambogin. Importantly, cambogin treatment led to the activation of activating transcription factor-2 (ATF-2) and the trimethylation of histone H3K9 in the activator protein 1 (AP-1) binding region of the Bcl-2 gene promoter. Finally, cambogin exhibited a potential antitumor effect in MCF-7 breast cancer xenografts without apparent toxicity. Taken in conjunction, the present study indicates that cambogin can induce breast adenocarcinoma cell apoptosis and therefore represents therapeutic potential for cancer treatment. Mol Cancer Ther; 14(7); 1738–49. ©2015 AACR.

Supplementary Figure 4. We studied the content of GSSG, TGSH and GSH in response to cambogin in MCF-7 cells. GSSG (oxidized glutathione) levels were elevated upon cambogin treatment whereas GSH levels were decreased from 4 to 24 h, which indicates possible ROS production in cambogin-treated MCF-7 cells.

Supplementary Figure 5. We have shown here that in MCF-7, MDA-MB-468, and SK-BR-3 cells, the pro-apoptotic effects of cambogin were prevented by antioxidative enzymes (CAT and SOD), further supporting the notion that ROS production is responsible for cambogin-induced cell death.

Supplementary Figure 2. Related to Fig. 2, the caspase-independent mitochondrial apoptotic pathway was also activated in response to cambogin in MDA-MB-468 and SK-BR-3 cells, similar to MCF-7 cells tested in Fig. 2.

Supplementary Figure 3. Related to Fig. 4, cambogin-induced apoptosis was ROS-dependent in MDA-MB-468 and SK-BR-3 cells, similar to MCF-7 cells tested in Fig. 4.

Supplementary Figure 6. Related to Fig.5, in MDA-MB-468 and SK-BR-3 cells, similar to MCF-7 cells tested in Fig. 5, JNK/SAPK signaling pathway is also activated upon cambogin treatment, and this is responsible for cambogin-induced cell death.

Supplementary Figure 1. Related to Fig.1, cambogin markedly inhibited the proliferation and colongenicity, as well as induced apoptosis in MDA-MB-468 and SK-BR-3 cells in a time- and dose-dependent fashion, similar to MCF-7 cells tested in Fig. 1.

Supplementary Information. Supplementary Material & Methods; supplementary figure legends; supplementary table 1 and reference.

Background: Previous research has identified a small subgroup of older adults that maintain a high level of cognitive functioning well into advanced age. Investigation of those with superior cognitive performance (SCP) for their age is important, as age-related decline has previously been thought to be inevitable. Objective: Preservation of cortical thickness and volume was evaluated in 76 older adults with SCP and 100 typical older adults (TOAs) assessed up to five times over six years. Methods: Regions of interest (ROIs) found to have been associated with super-aging status (a construct similar to SCP status) in previous literature were investigated, followed by a discovery phase analyses of additional regions. SCPs were aged 70 + at baseline, scoring at/above normative memory (CVLT-II) levels for demographically similar individuals aged 30–44 years old, and in the unimpaired range for all other cognitive domains over the course of the study. Results: In linear mixed models, following adjustment for multiple comparisons, there were no significant differences between rates of thinning or volume atrophy between SCPs and TOAs in previously identified ROIs, or the discovery phase analyses. With only amyloid-β negative individuals in the analyses, again there were no significant differences between SCPs and TOAs. Conclusion: The increased methodological rigor in classifying groups, together with the influence of cognitive reserve, are discussed as potential factors accounting for our findings as compared to the extant literature on those with superior cognitive performance for their age.

PURPOSE The purpose of this essay is to improve computer-aided diagnosis of lung diseases by the removal of bone structures imagery such as ribs and clavicles, which may shadow a clinical view of lesions. This paper aims to develop an algorithm to suppress the imaging of bone structures within clinical x-ray images, leaving a residual portrayal of lung tissue; such that these images can be used to better serve applications, such as lung nodule detection or pathology based on the radiological reading of chest x rays. METHODS We propose a conditional Adversarial Generative Network (cGAN) (Mirza and Osindero, Conditional generative adversarial nets, 2014.) model, consisting of a generator and a discriminator, for the task of bone shadow suppression. The proposed model utilizes convolutional operations to expand the size of the receptive field of the generator without losing contextual information while downsampling the image. It is trained by enforcing both the pixel-wise intensity similarity and the semantic-level visual similarity between the generated x-ray images and the ground truth, via optimizing an L-1 loss of the pixel intensity values on the generator side and a feature matching loss on the discriminator side, respectively. RESULTS The framework we propose is trained and tested on an open-access chest radiograph dataset for benchmark. Results show that our model is capable of generating bone-suppressed images of outstanding quality with a limited number of training samples (N = 272). CONCLUSIONS Our approach outperforms current state-of-the-art bone suppression methods using x-ray images. Instead of simply downsampling images at different scales, our proposed method mitigates the loss of contextual information by utilizing dilated convolutions, which gains a noticeable quality improvement for the outputs. On the other hand, our experiment shows that enforcing the semantic similarity between the generated and the ground truth images assists the adversarial training process and achieves better perceptual quality.

Background/Objective: Hepcidin, an iron-regulating hormone, suppresses the release of iron by binding to the iron exporter protein, ferroportin, resulting in intracellular iron accumulation. Given that iron dyshomeostasis has been observed in Alzheimer’s disease (AD) together with elevated serum hepcidin levels, the current study examined whether elevated serum hepcidin levels are an early event in AD pathogenesis by measuring the hormone in cognitively normal older adults at risk of AD, based on high neocortical amyloid-β load (NAL). Methods: Serum hepcidin levels in cognitively normal participants (n = 100) aged between 65–90 years were measured using ELISA. To evaluate NAL, all participants underwent 18F-florbetaben positron emission tomography. A standard uptake value ratio (SUVR)

Background: Non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutations will inevitably develop drug resistance after being treated with the third-generation EGFR-tyrosine kinase inhibitor (TKI), osimertinib. In recent years, many studies have been focusing on the ability of exosomal miRNAs secreted by tumor cells to transmit resistance information. However, the mechanism of exosome-derived miRNAs in osimertinib resistance remains unexplored.Methods: We extracted and sequenced exosomes from the supernatant of the osimertinib-resistant cell line, H1975-OR, and the sensitive cell line, H1975. The results were compared with plasma exosome sequencing before and after the appearance of drug resistance in three NSCLC clinical patients treated with oral osimertinib. Exosome-derived miRNAs that had significantly increased expression levels after osimertinib resistance were screened for expanded validation in other 64 NSCLC patients.Results: Cluster analysis of the target genes revealed that exosomal miRNAs participate in osimertinib resistance mechanisms through the activation of bypass pathways (RAS-MAPK pathway abnormality and PI3K pathway activation). Exosome-derived miR-184 (p = 0.0325) and miR-3913-5p (p = 0.0169) expression levels increased significantly after the onset of osimertinib resistance. Exosomal miR-184 was correlated with lactate dehydrogenase levels (p = 0.018). Exosomal miR-3913-5p was associated with TNM stage (p = 0.045), platelet count (p = 0.024), tumor marker carcinoembryonic antigen (p = 0.045), and distant metastases (p = 0.049), especially bone metastasis (p = 0.03). In the subgroup of patients with EGFR exon 21 L858R point mutation, miR-184 (p = 0.0104) and miR-3913-5p (p = 0.0085) derived from serum exosomes had both significantly increased expression levels. In the subgroup of T790M-positive patients, miR-3913-5p derived from serum exosomes may also be a good indicator of osimertinib resistance (p = 0.013).Conclusions: The expression levels of miR-184 and miR-3913-5p derived from exosomes in the peripheral blood of NSCLC patients could be used as biomarkers to indicate osimertinib resistance.

Cellular senescence is one of the most significant factors involved in aging and age-related diseases. Senescence of vascular smooth muscle cells (VSMCs) adversely affects the function of the cardiovascular system and contributes to the development of atherosclerosis, hypertension, and other cardiovascular diseases. Glucagon-like peptide-1 (GLP-1) is an important incretin hormone involved in insulin release and vascular tone. GLP-1 is quickly degraded by the enzyme dipeptidyl peptidase-4 (DPP-4). Omarigliptin is a new DPP-4 inhibitor that has demonstrated anti-inflammatory and antioxidative stress properties. In the present study, we investigated the effects of the selective DPP-4 inhibitor omarigliptin (OMG) on VSMCs exposed to insult from tumor necrosis factor-α (TNF-α), one of the main inflammatory signaling molecules involved in cellular senescence. We found that OMG could suppress TNF-α-induced expression of pro-inflammatory cytokines (interleukin-1β (IL-1β), IL-6, and IL-8) and inhibit oxidative stress by reducing the production of H2O2 and protein carbonyl. OMG ameliorated the increase in senescence-associated β-galactosidase (SA-β-gal) and telomerase activity induced by TNF-α. The plasminogen activator inhibitor-1 (PAI-1)/p53/p21 pathway is a key inducer of cellular senescence. OMG ameliorated the acetylation of p53 at lysine 382 (K382) and subsequent activation of p21 via inhibition of PAI-1. Importantly, our experiments revealed that blockage of silent information-regulator 1 (SIRT1) abolished the inhibitory effects of OMG on p53 acetylation, SA-β-gal activity, and telomerase activity in VSMCs. These results suggest that OMG may have the potential to delay or prevent the progression of age-related cardiovascular diseases by modulating the activity of SIRT1.

Hepatocellular carcinoma (HCC) is an aggressive malignancy with increasing mortality in China. Angiogenesis is crucial for tumor formation, development and metastasis in HCC. Previous studies indicated that high expression levels of elongation factor 2 kinase (eEF2K), a protein kinase that negatively regulates the elongation stage of translation, were associated with poor prognosis of HCC. Here, we show that pharmacological inhibition or knockdown of eEF2K in highly metastatic liver cancer cells inhibits their colony forming and migratory capacities, as well as reducing their invasiveness. Importantly, knocking down eEF2K by lentiviral directed shRNA prevented tumor growth and angiogenesis of HCC in mice. Silencing of eEF2K in endothelial cells (HUVECs) led to a reduction in vascularization, evidenced by a decrease in capillary-like structures in the matrigel. Notably, knocking down eEF2K reduced the expression of angiogenesis-related growth factors in liver cancer cells and the expression of growth factor receptors on HUVECs, and thus restricted signaling crosstalk that promotes angiogenesis between HCC cells and endothelial cells. We also showed that silencing of eEF2K effectively reduced protein levels of SP1/KLF5 transcription factors and hence decreased the levels of bound SP1/KLF5 to the VEGF promoter, resulted in a decrease in VEGF mRNA expression. Knocking down eEF2K also led to a striking decrease in the phosphorylation of PI3K/Akt and STAT3, indicating inactivation of these tumorigenic pathways. Taken together, our data suggest that eEF2K contributes to angiogenesis and tumor progression in HCC via SP1/KLF5-mediated VEGF expression, as well as the subsequent stimulation of PI3K/Akt and STAT3 signaling.

Background Neutrophil-to-lymphocyte ratio (NLR) is related to prognosis in non-small cell lung cancer (NSCLC). However, no consensus on the relationship of pretreatment NLR and survival outcomes of systemic therapy in NSCLC exists. This meta-analysis investigated the prognostic role of pretreatment NLR during systemic therapy for NSCLC, including chemotherapy, immunotherapy and targeted therapy. Methods PubMed, Web of Science and Cochrane Library databases were systematically searched up to April 09, 2019. Hazard ratios (HRs) with their 95% confidence intervals (CIs) were pooled to investigate the association of pretreatment NLR with progression-free survival (PFS) and overall survival (OS). Results In total, 27 articles with 4,298 participants were selected. The pooled results showed that elevated pretreatment NLR was associated with inferior PFS (HR, 1.45, 95% CI, 1.28-1.66) and OS (HR, 1.63, 95% CI, 1.43-1.84) during systemic therapy. Subgroup analyses according to the treatment strategy suggested that higher pretreatment NLR was significantly associated with shorter survival in all therapies, including chemotherapy (PFS HR, 1.74, 95% CI, 1.39-2.17; OS HR, 1.73, 95% CI, 1.26-2.36), immunotherapy (PFS HR, 1.53, 95% CI, 1.27-1.84; OS HR, 2.50, 95% CI, 1.60-3.89) and targeted therapy (PFS HR, 1.53, 95% CI, 1.04-2.25; OS HR, 1.92, 95% CI, 1.14-3.24). Conclusions Pretreatment NLR is a promising prognostic indicator for NSCLC patients receiving systemic therapy, including chemotherapy, immunotherapy and targeted therapy.

Immune checkpoint inhibitors (ICIs) combined with chemotherapy have been applied as a first-line treatment for lung cancer, but consistent beneficial results have not been documented. Therefore, our meta-analysis aimed to evaluate the effectiveness and safety of combination therapy to promote its application.We searched electronic databases for studies that estimated the safety and efficacy of combined therapy. The objective response rate (ORR) and disease control response (DCR) parameters were evaluated with odds ratio (OR) values of the combination arm over the non-combination arm. Hazard ratios (HR) and its 95% confidence intervals (95% CI) were used to calculate progression-free survival (PFS) and overall survival (OS) in the combination and non-combination arms. All treatment-related adverse events (TRAEs) and 3 to 5 TRAEs were expressed as relative risk (RR) values of the combination arm over the non-combination arm.Ten eligible studies involving 4,887 patients were identified. The pooled ORs for ORR and DCR were 1.85 (95% CI: 1.30-2.63, P0.01) and 1.14 (95% CI: 0.70-1.86, P0.01), respectively. The pooled HRs for PFS and OS were 0.67 (95% CI: 0.58-0.79, P0.001) and 0.76 (95% CI: 0.65-0.88, P0.001), respectively. In subgroup analysis, ORR and DCR were significantly improved in the programmed cell death-1/L1 (PD-1/L1) blockade for non-small cell lung cancer (NSCLC) group (subgroup A), with a combined OR values of 2.36 (95% CI: 1.79-3.13, P0.001) and 1.92 (95% CI: 1.10-3.35, P0.001), respectively. However, no significant benefits were observed in the cytotoxic T lymphocyte antigen-4 (CTLA-4) blockade for small cell lung cancer (SCLC) (subgroup B) and CTLA-4 blockade for NSCLC groups (subgroup C). In addition, a significant improvement in PFS was observed in subgroup A, subgroup B and subgroup C, with pooled HR values of 0.58 (95% CI: 0.52-0.63, P0.001), 0.86 (95% CI: 0.76-0.97, P0.05) and 0.83 (95% CI: 0.68-1.00, P0.05), respectively. Only subgroup A exhibited an OS benefit, with a combined HR value of 0.67 (95% CI: 0.55-0.81, P0.001). Moreover, as the expression of PD-L1 increased, the PFS and OS benefits were more significantly. Furthermore, patients without central nervous system (CNS) metastasis who were treated with PD-1/L1 inhibitors had a longer OS than patients with CNS metastasis (HR: 0.67, 95% CI: 0.55-0.80, P0.001). Finally, combined therapy was associated with 3 to 5 TRAEs (RR: 1.26, 95% CI: 1.08-1.47; P0.01).Patients treated with immunotherapy and chemotherapy in combination exhibited superior in ORR, DCR, PFS and OS as well as slightly increased TRAE levels compared with those of patients treated with either monotherapy.

Background The third-generation of EGFR-TKI Osimertinib has become an important treatment option for patients with EGFR-mutant advanced NSCLC. In recent years, more and more studies have begun to pay attention to the ability of miRNAs in exosomes secreted by tumor cells to transmit resistance information. The mechanisms of exosomal miRNAs involved in Osimertinib resistance need to be studied. Methods We constructed an Osimertinib-resistant cell line H1975-OR, and collected the exosomes from the cells of the drug-resistant strain and the sensitive strain, and extracted respective RNA for sequencing. We also compared miRNAs in serum exosomes of three patients before and after resistance to Osimertinib. Enlarged samples were then validated in 64 NSCLC patients. Results Cluster analysis of target genes revealed that miRNAs in exosomes participate in Osimertinib resistance mechanisms through the activation of bypass pathways (RAS-MAPK pathway abnormality, PI3K pathway activation). MiR-184 and miR-3913-5p increased significantly in serum exosomes of patients with later Osimertinib resistance. These two miRNAs mainly cause EGFR resistance in two types of NSCLC patients, EGFR exon 21 L858R mutation and T790M positive. Conclusions Exosomes miR-184 and miR-3913-5p mainly cause Osimertinib resistance in lung cancer patients containing EGFR exon 21 L858R+ and T790M+. They are regarded as important biomarkers of third-generation EGFR-TKI resistance. This not only enriches the application of liquid biopsy in lung cancer drug resistance, but also provides a direction for future targeted drug research.

BACKGROUND: Exudative pleural effusion (EPE) is a common diagnostic challenge. The utility of medical thoracoscopy (MT) and closed pleural biopsy (CPB) to aid in the diagnosis of EPE has been reported in many published studies. Herein, we perform a systematic review and meta-analysis to compare the diagnostic yield and safety of CPB and MT in EPE. METHODS: Four databases were searched for studies reporting the diagnostic yield of CPB and MT for EPE. The quality of the included studies was evaluated according to the quality assessment of diagnostic accuracy studies (QUADAS) tool. The pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), and complication risks were compared between the two groups. RESULTS: Ten studies dealing with CPB and twenty-three studies dealing with MT for the diagnosis of EPE were included in this meta-analysis. Pooled sensitivity, specificity, PLR, NLR and DOR of CPB group was 77%, 99%, 32.55, 0.22, 165.71, respectively, while pooled sensitivity, specificity, PLR, NLR and DOR of MT group was 93%, 100%, 10.82, 0.08, 162.81, respectively. The area under the summary receiver operating characteristic (SROC) curve of CPB and MT were both 0.97. The ability of CPB to diagnose non-malignant diseases was like MT (69% vs. 68%), while the ability was lower than that of MT to diagnose malignant diseases (72% vs. 92%). The pooled diagnostic accuracy of CPB and MT for mesothelioma was 26% (95% CI, 14–38%) and 42% (95% CI, 22–62%) (P

Background Heme and iron homeostasis is perturbed in Alzheimer’s disease (AD); therefore, the aim of the study was to examine the levels and association of heme with iron-binding plasma proteins in cognitively normal (CN), mild cognitive impairment (MCI), and AD individuals from the Australian Imaging, Biomarker and Lifestyle Flagship Study of Ageing (AIBL) and Kerr Anglican Retirement Village Initiative in Ageing Health (KARVIAH) cohorts. Methods Non-targeted proteomic analysis by high-resolution mass spectrometry was performed to quantify relative protein abundances in plasma samples from 144 CN individuals from the AIBL and 94 CN from KARVIAH cohorts and 21 MCI and 25 AD from AIBL cohort. ANCOVA models were utilized to assess the differences in plasma proteins implicated in heme/iron metabolism, while multiple regression modeling (and partial correlation) was performed to examine the association between heme and iron proteins, structural neuroimaging, and cognitive measures. Results Of the plasma proteins implicated in iron and heme metabolism, hemoglobin subunit β (p = 0.001) was significantly increased in AD compared to CN individuals. Multiple regression modeling adjusted for age, sex, APOEε4 genotype, and disease status in the AIBL cohort revealed lower levels of transferrin but higher levels of hemopexin associated with augmented brain amyloid deposition. Meanwhile, transferrin was positively associated with hippocampal volume and MMSE performance, and hemopexin was negatively associated with CDR scores. Partial correlation analysis revealed lack of significant associations between heme/iron proteins in the CN individuals progressing to cognitive impairment. Conclusions In conclusion, heme and iron dyshomeostasis appears to be a feature of AD. The causal relationship between heme/iron metabolism and AD warrants further investigation.

Platelets are implicated as key players in the metastatic dissemination of tumor cells. Previous evidence demonstrated platelets retained cytoplasmic RNAs with physiologically activity, splicing pre-mRNA to mRNA and translating into functional proteins in response to external stimulation. Recently, platelets gene profile of healthy or diseased individuals were characterized with the help of RNA sequencing (RNA-Seq) in some studies, leading to new insights into the mechanisms underlying disease pathogenesis. In this study, we performed RNA-seq in platelets from 7 healthy individuals and 15 non-small cell lung cancer (NSCLC) patients. Our data revealed a subset of near universal differently expressed gene (DEG) profiles in platelets of metastatic NSCLC compared to healthy individuals, including 626 up-regulated RNAs (mRNAs and ncRNAs) and 1497 down-regulated genes. The significant over-expressed genes showed enrichment in focal adhesion, platelets activation, gap junction and adherens junction pathways. The DEGs also included previously reported tumor-related genes such as PDGFR, VEGF, EGF, etc., verifying the consistence and significance of platelet RNA-Seq in oncology study. We also validated several up-regulated DEGs involved in tumor cell-induced platelet aggregation (TCIPA) and tumorigenesis. Additionally, transcriptomic comparison analyses of NSCLC subgroups were conducted. Between non-metastatic and metastatic NSCLC patients, 526 platelet DEGs were identified with the most altered expression. The outcomes from subgroup analysis between lung adenocarcinoma and lung squamous cell carcinoma demonstrated the diagnostic potential of platelet RNA-Seq on distinguishing tumor histological types.

eIF4E plays key roles in protein synthesis and tumorigenesis. It is phosphorylated by the kinases MNK1 and MNK2. Binding of MNKs to eIF4G enhances their ability to phosphorylate eIF4E. Here, we show that mTORC1, a key regulator of mRNA translation and oncogenesis, directly phosphorylates MNK2 on Ser74. This suppresses MNK2 activity and impairs binding of MNK2 to eIF4G. These effects provide a novel mechanism by which mTORC1 signaling impairs the function of MNK2 and thereby decreases eIF4E phosphorylation. MNK2[S74A] knock-in cells show enhanced phosphorylation of eIF4E and S6K1 (i.e., increased mTORC1 signaling), enlarged cell size, and increased invasive and transformative capacities. MNK2[Ser74] phosphorylation was inversely correlated with disease progression in human prostate tumors. MNK inhibition exerted anti-proliferative effects in prostate cancer cells in vitro. These findings define a novel feedback loop whereby mTORC1 represses MNK2 activity and oncogenic signaling through eIF4E phosphorylation, allowing reciprocal regulation of these two oncogenic pathways. Refereed/Peer-reviewed

BackgroundBlood markers indicative of neurodegeneration (neurofilament light chain; NFL), Alzheimer’s disease amyloid pathology (amyloid-β; Aβ), and neuroinflammation (kynurenine pathway; KP metabolites) have been investigated independently in neurodegenerative diseases. However, the association of these markers of neurodegeneration and AD pathology with neuroinflammation has not been investigated previously. Therefore, the current study examined whether NFL and Aβ correlate with KP metabolites in elderly individuals to provide insight on the association between blood indicators of neurodegeneration and neuroinflammation.MethodsCorrelations between KP metabolites, measured using liquid chromatography and gas chromatography coupled with mass spectrometry, and plasma NFL and Aβ concentrations, measured using single molecule array (Simoa) assays, were investigated in elderly individuals aged 65–90 years, with normal global cognition (Mini-Mental State Examination Score ≥ 26) from the Kerr Anglican Retirement Village Initiative in Ageing Health cohort.ResultsA positive correlation between NFL and the kynurenine to tryptophan ratio (K/T) reflecting indoleamine 2,3-dioxygenase activity was observed (r = .451,p r = .364,p r = .384,p r = .246,p = .014), anthranilic acid (r = .311,p = .002), and quinolinic acid (r = .296,p = .003). Further, significant associations were observed between plasma Aβ40 and the K/T (r = .375,p r = .374,p r = .352,p r = .381,p r = .352,p r = .215,p = .034), kynurenic acid (r = .214,p = .035), anthranilic acid (r = .278,p = .006), and quinolinic acid (r = .224,p = .027) in the cohort. On stratifying participants based on their neocortical Aβ load (NAL) status, NFL correlated with KP metabolites irrespective of NAL status; however, associations between plasma Aβ and KP metabolites were only pronounced in individuals with high NAL while associations in individuals with low NAL were nearly absent.ConclusionsThe current study shows that KP metabolite changes are associated with biomarker evidence of neurodegeneration. Additionally, the association between KP metabolites and plasma Aβ seems to be NAL status dependent. Finally, the current study suggests that an association between neurodegeneration and neuroinflammation manifests in the periphery, suggesting that preventing cytoskeleton cytotoxicity by KP metabolites may have therapeutic potential.

Eukaryotic elongation factor 2 kinase (eEF2K) negatively regulates the elongation phase of mRNA translation and hence protein synthesis. Increasing evidence indicates that eEF2K plays an important role in the survival and migration of cancer cells and in tumor progression. As demonstrated by two-dimensional wound-healing and three-dimensional transwell invasion assays, knocking down or inhibiting eEF2K in cancer cells impairs migration and invasion of cancer cells. Conversely, exogenous expression of eEF2K or knocking down eEF2 (the substrate of eEF2K) accelerates wound healing and invasion. Importantly, using LC-HDMSE analysis, we identify 150 proteins whose expression is decreased and 73 proteins which are increased upon knocking down eEF2K in human lung carcinoma cells. Of interest, 34 downregulated proteins are integrins and other proteins implicated in cell migration, suggesting that inhibiting eEF2K may help prevent cancer cell mobility and metastasis. Interestingly, eEF2K promotes the association of integrin mRNAs with polysomes, providing a mechanism by which eEF2K may enhance their cellular levels. Consistent with this, genetic knock down or pharmacological inhibition of eEF2K reduces the protein expression levels of integrins. Notably, pharmacological or genetic inhibition of eEF2K almost completely blocked tumor growth and effectively prevented the spread of tumor cells in vivo. High levels of eEF2K expression were associated with invasive carcinoma and metastatic tumors. These data provide the evidence that eEF2K is a new potential therapeutic target for preventing tumor metastasis.

Ferritin, an iron storage and regulation protein, has been associated with Alzheimer's disease (AD); however, it has not been investigated in preclinical AD, detected by neocortical amyloid-β load (NAL), before cognitive impairment. Cross-sectional analyses were carried out for plasma and serum ferritin in participants in the Kerr Anglican Retirement Village Initiative in Aging Health cohort. Subjects were aged 65-90 years and were categorized into high and low NAL groups via positron emission tomography using a standard uptake value ratio cutoff=1.35. Ferritin was significantly elevated in participants with high NAL compared with those with low NAL, adjusted for covariates age, sex, apolipoprotein E ɛ4 carriage and levels of C-reactive protein (an inflammation marker). Ferritin was also observed to correlate positively with NAL. A receiver operating characteristic curve based on a logistic regression of the same covariates, the base model, distinguished high from low NAL (area under the curve (AUC)=0.766), but was outperformed when plasma ferritin was added to the base model (AUC=0.810), such that at 75% sensitivity, the specificity increased from 62 to 71% on adding ferritin to the base model, indicating that ferritin is a statistically significant additional predictor of NAL over and above the base model. However, ferritin's contribution alone is relatively minor compared with the base model. The current findings suggest that impaired iron mobilization is an early event in AD pathogenesis. Observations from the present study highlight ferritin's potential to contribute to a blood biomarker panel for preclinical AD.

Advanced connectivity studies in toddlers with Autism Spectrum Disorder (ASD) are increasing and consistently reporting a disruption of brain connectivity. However, most of these studies compare ASD and typically developing subjects, thus providing little information on the specificity of the abnormalities detected in comparison with other developmental disorders (other-DD). We recruited subjects aged below 36 months who received a clinical diagnosis of Neurodevelopmental Disorder (32 ASD and 16 other-DD including intellectual disability and language disorder) according to DSM-IV TR. Structural and diffusion MRI were acquired to perform whole brain probabilistic and anatomically constrained tractography. Network connectivity matrices were built encoding the number of streamlines (DNUM ) and the tract-averaged fractional anisotropy (DFA ) values connecting each pair of cortical and subcortical regions. Network Based Statistics (NBS) was finally applied on the connectivity matrices to evaluate the network differences between the ASD and other-DD groups. The network differences resulted in an over-connectivity pattern (i.e., higher DNUM and DFA values) in the ASD group with a significance of P

Understanding the relationships between the structure and function of the brain largely relies on the qualitative assessment of Magnetic Resonance Images (MRIs) by expert clinicians. Automated analysis systems can support these assessments by providing quantitative measures of brain injury. However, the assessment of deep gray matter structures, which are critical to motor and executive function, remains difficult as a result of large anatomical injuries commonly observed in children with Cerebral Palsy (CP). Hence, this article proposes a robust surrogate marker of the extent of deep gray matter injury based on impingement due to local ventricular enlargement on surrounding anatomy. Local enlargement was computed using a statistical shape model of the lateral ventricles constructed from 44 healthy subjects. Measures of injury on 95 age-matched CP patients were used to train a regression model to predict six clinical measures of function. The robustness of identifying ventricular enlargement was demonstrated by an area under the curve of 0.91 when tested against a dichotomised expert clinical assessment. The measures also showed strong and significant relationships for multiple clinical scores, including: motor function (r = 0.62, P < 0.005), executive function (r = 0.55, P < 0.005), and communication (r = 0.50, P < 0.005), especially compared to using volumes obtained from standard anatomical segmentation approaches. The lack of reliance on accurate anatomical segmentations and its resulting robustness to large anatomical variations is a key feature of the proposed automated approach. This coupled with its strong correlation with clinically meaningful scores, signifies the potential utility to repeatedly assess MRIs for clinicians diagnosing children with CP. Hum Brain Mapp 37:3795–3809, 2016.

Background: Aberrant amyloid-β (Aβ) deposition in the brain occurs two decades prior to the manifestation of Alzheimer’s disease (AD) clinical symptoms and therefore brain Aβ load measured using PET serves as a gold standard biomarker for the early diagnosis of AD. However, the uneconomical nature of PET makes blood markers, that reflect brain Aβ deposition, attractive candidates for investigation as surrogate markers. Objective:Investigation of plasma Aβ as a surrogate marker for brain Aβ deposition in cognitively normal elderly individuals. Methods: Plasma Aβ40 and Aβ42 concentrations were measured using the ultrasensitive Single Molecule Array (Simoa) assay in 95 cognitively normal elderly individuals, who have all undergone PET to assess brain Aβ deposition. Based on the standard uptake value ratios (SUVR) obtained from PET imaging, using the tracer 18F-Florbetaben, plasma Aβ was compared between 32 participants assessed to have low brain Aβ load (Aβ–, SUVR

Executive function (EF) is a set of cognitive capabilities considered essential for successful daily living, and is negatively affected by ageing and neurodegenerative conditions. Underpinning EF performance are functional nodes in the executive control network (ECN), while the structural connectivity underlying this network is not well understood. In this paper, we evaluated the structural white matter tracts that interconnect the ECN and investigated their relationship to the EF performance. Using high‐angular resolution diffusion MRI data, we performed tractography analysis of structural connectivity in a cognitively normal cohort (n = 140), specifically targeting the connectivity between ECN nodes. Our data revealed the presence of a strongly‐connected “structural core” of the ECN comprising three components: interhemispheric frontal connections, a fronto‐parietal subnetwork and fronto‐striatal connections between right dorsolateral prefrontal cortex and right caudate. These pathways were strongly correlated with EF performance (p = .003). Post‐hoc analysis of subregions within the significant ECN connections showed that these effects were driven by a highly specific subset of interconnected cortical regions. The structural core subnetwork of the functional ECN may be an important feature crucial to a better future understanding of human cognition and behaviour.

Long non-coding RNAs (lncRNAs) have been identified to serve an important role in the occurrence, development and metastasis of tumours. However, the role of linc00467 in lung adenocarcinoma (LAD) is unclear. In the present study, it was demonstrated that linc00467 expression was upregulated in human lung tumour tissues compared with normal tissues. In addition, high levels of linc00467 expression were associated with larger tumour sizes and later TNM stages. Functional experiments suggested that linc00467 promoted LAD cell proliferation, migration and invasion, and inhibited apoptosis in vitro. Knockdown of linc00467 altered the expression of downstream genes, including HtrA serine peptidase 3 (HTRA3), and RNA immunoprecipitation and chromatin immunoprecipitation assays indicated that linc00467 recruited EZH2 to the HTRA3 promoter to inhibit its expression. Taken together, the results of the present study indicated that linc00467 served an oncogenic role in LAD tumourigenesis, suggesting that it may be used as a novel diagnostic biomarker and therapeutic target for LAD.

Personality factors have been associated with Alzheimer’s disease (AD) and dementia, but they have not been examined against markers of regional brain glucose metabolism (a primary measure of brain functioning) in older adults without clinically diagnosed cognitive impairment. The relationship between personality factors derived from the five-factor model and cerebral glucose metabolism determined using positron emission tomography (PET) with [18F]-2-fluoro-2-deoxy-d-glucose (18F-FDG-PET) was examined in a cohort of 237 non-demented, community-dwelling older adults aged 60–89 years (M ± SD = 73.76 ± 6.73). Higher neuroticism and lower scores on extraversion and conscientiousness were significantly associated with decreased glucose metabolism in brain regions typically affected by AD neuropathological processes, including the hippocampus and entorhinal cortex. Furthermore, while there were significant differences between apolipoprotein E (APOE) ε4 allele carriers and non-carriers on 18F-FDG-PET results in the neocortex and other brain regions (p

Table S1. Cohort characteristics based on neurofilament light chain quartiles. (DOCX 15 kb)

The purpose of this research was to explore the mechanism of action of B. cereus on Pengze crucian carp (Carassius auratus var. Pengze). Fish fed basal diet with supplementation of living (LBC group) and heated-inactivated (HIBC group) B. cereus at the dose of 1 × 109 CFU kg−1, and basal diet served as Control group. Our results indicated that both additives remarkably promoted the growth of fish (P

// Kaikai Shen 1, * , Zhichao Xi 1, * , Jianling Xie 4 , Hua Wang 5 , Chanlu Xie 6 , C.Soon Lee 6, 7 , Paul Fahey 6 , Qihan Dong 6, 7, 8 , Hongxi Xu 1, 2, 3 1 School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai, China 2 Institute of Arthritis Research, Shanghai Academy of Chinese Medical Sciences, Guanghua Integrative Medicine Hospital/ Shanghai University of T.C.M, Shanghai, China 3 Engineering Research Center of Shanghai Colleges for TCM New Drug Discovery, Shanghai, China 4 Nutrition and Metabolism, South Australian Health and Medical Research Institute, Adelaide, Australia 5 Stanley Ho Center for Emerging Infectious Diseases, The Chinese University of Hong Kong, Hong Kong, China 6 School of Science and Health, The University of Western Sydney, Sydney, Australia 7 Central Clinical School and Bosch Institute, The University of Sydney, Sydney, Australia 8 Department of Endocrinology, Royal Prince Alfred Hospital, Sydney, Australia * These authors contributed equally to this work Correspondence to: Hongxi Xu, email: xuhongxi88@gmail.com Qihan Dong, email: q.dong@uws.edu.au Keywords: hepatocellular carcinoma, cancer metastasis, Guttiferone K, profilin1, actin Received: November 20, 2015 Accepted: July 18, 2016 Published: August 01, 2016 ABSTRACT Hepatocellular carcinoma (HCC) is an aggressive malignancy and the 5-year survival rate of advanced HCC is < 10%. Guttiferone K (GUTK) isolated from the Garcinia genus inhibited HCC cells migration and invasion in vitro and metastasis in vivo without apparent toxicity. Proteomic analysis revealed that actin-binding protein profilin 1 (PFN1) was markedly increased in the presence of GUTK. Over-expression of PFN1 mimicked the effect of GUTK on HCC cell motility and metastasis. The effect of GUTK on cell motility was diminished when PFN1 was over-expressed or silenced. Over-expression of PFN1 or incubation with GUTK decreased F-actin levels and the expression of proteins involved in actin nucleation, branching and polymerization. Moreover, a reduction of PFN1 protein levels was common in advanced human HCC and associated with poor survival rate. In conclusion, GUTK effectively suppresses the motility and metastasis of HCC cells mainly by restoration of aberrantly reduced PFN1 protein expression.

// Kaikai Shen 1, * , Fangfang Lu 1, * , Jianling Xie 4, 5 , Minfeng Wu 1 , Bo Cai 1 , Yurong Liu 1 , Hong Zhang 1, 3 , Hongsheng Tan 1, 3 , Yingyi Pan 1 , Hongxi Xu 1, 2, 3 1 School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China 2 Institute of Arthritis Research, Shanghai Academy of Chinese Medical Sciences, Guanghua Integrative Medicine Hospital/Shanghai University of T.C.M, Shanghai 201203, China 3 Engineering Research Center of Shanghai Colleges for TCM New Drug Discovery, Shanghai 201203, China 4 Nutrition & Metabolism, South Australian Health & Medical Research Institute, North Terrace, Adelaide SA5000, Australia 5 Centre for Biological Sciences, Life Sciences Building, University of Southampton, Southampton, SO17 1BJ, UK * These authors have contributed equally to this work Correspondence to: Hongxi Xu, email: xuhongxi88@gmail.com Yingyi Pan, email: pyy16@126.com Keywords: breast cancer, cambogin, reactive oxygen species, NADPH oxidase 1, thioredoxin-1/ASK1 complex Received: December 02, 2015 Accepted: June 29, 2016 Published: July 13, 2016 ABSTRACT Cambogin, a bioactive polycyclic polyprenylated acylphoroglucinol (PPAP) derived from the Garcinia genus, possesses proapoptotic effect in medulloblastoma and breast cancer cells. We have previously demonstrated that the proapoptotic effect of cambogin is driven by the production of reactive oxygen species (ROS). Here we have shown that the inhibitory effect of cambogin on cell proliferation is associated with the loss of mitochondrial transmembrane potential (Δ Ψ m ) and mitochondrial fragmentation. Cambogin also promotes the mutual complex formation of the membrane-bound subunit p22 phox of NADPH oxidase 1 (NOX1), as well as the phosphorylation of the cytosolic subunit p47 phox , subsequently enhancing membrane-bound NOX1 activity, which leads to increases in intracellular and mitochondrial levels of O 2 .- and H 2 O 2 . Pharmacological inhibition of NOX1 using apocynin (pan-NOX inhibitor), ML171 (NOX1 inhibitor) or siRNA against NOX1 prevents the increases in O 2 .- and H 2 O 2 levels and the anti-proliferative effect of cambogin. Antioxidants, including SOD (superoxide dismutase), CAT (catalase) and EUK-8, are also able to restore cell viability in the presence of cambogin. Besides, cambogin increases the dissociation of thioredoxin-1 (Trx1) from ASK1, switching the inactive form of ASK1 to the active kinase, subsequently leads to the phosphorylation of JNK/SAPK, which is abolished upon ML171 treatment. The proapoptotic effect of cambogin in breast cancer cells is also aggravated upon knocking down Trx1 in MCF-7 cells. Taken in conjunction, these data indicate that the anti-proliferative and pro-apoptotic effect of cambogin is mediated via inducing NOX1-dependent ROS production and the dissociation of ASK1 and Trx1.

The aim of this study is to investigate the genetic influence on the cerebral cortex, based on the analyses of heritability and genetic correlation between grey matter (GM) thickness, derived from structural MR images (sMRI), and associated white matter (WM) connections obtained from diffusion MRI (dMRI). We measured on sMRI the cortical thickness (CT) from a large twin imaging cohort using a surface-based approach (N = 308, average age 22.8 ± 2.3 SD). An ACE model was employed to compute the heritability of CT. WM connections were estimated based on probabilistic tractography using fiber orientation distributions (FOD) from dMRI. We then fitted the ACE model to estimate the heritability of CT and FOD peak measures along WM fiber tracts. The WM fiber tracts where genetic influence was detected were mapped onto the cortical surface. Bivariate genetic modeling was performed to estimate the cross-trait genetic correlation between the CT and the FOD-based connectivity of the tracts associated with the cortical regions. We found some cortical regions displaying heritable and genetically correlated GM thickness and WM connectivity, forming networks under stronger genetic influence. Significant heritability and genetic correlations between the CT and WM connectivity were found in regions including the right postcentral gyrus, left posterior cingulate gyrus, right middle temporal gyri, suggesting common genetic factors influencing both GM and WM. Hum Brain Mapp 37:2331-2347, 2016. © 2016 Wiley Periodicals, Inc.

This study was aimed to evaluate the effects of dietary supplementation of probiotics Bacillus cereus (BC), B. subtilis (BS), Paracoccus marcusii (PM), and Lactobacillus plantarum (LP) on growth, immune response, antioxidant property, and intestine health of juvenile grass carp. Our results showed that dietary probiotics could promote the growth performance, which was significantly increased in BS, PM, and LP groups compared to the fish in Control group. Also, probiotics supplementation significantly enhanced the immunity of fish measured by the content of C3 and C4, and/or AKP activity in serum, and elevated antioxidant property of intestine accompanying by a significantly lower content of malondialdehyde. Compared to Control group, dietary B. cereus and B. subtilis significantly affected intestinal antioxidant system through Nrf2 signaling pathway by up-regulating the expression of catalase, CuZn superoxide dismutase and/or Mn superoxide dismutase, and further improved the intestinal barrier function by up-regulating mRNA levels of ZO-1, ZO-2, ZO-3, and Clauding-12. Besides, B. cereus, P. marcusii, and L. plantarum supplementation could decrease intestinal inflammation by suppressing TLR4 signaling pathway, meanwhile P. marcusii supplementation up-regulated the expression of ZO-2 and ZO-3. Overall, our results indicated that dietary B. cereus, B. subtilis, P. marcusii, and L. plantarum had a positive impact on the growth performance, immunity and antioxidant property of grass carp, and B. cereus, B. subtilis, and P. marcusii exerted benefits on the intestinal health through elevating antioxidant property and decreasing permeability of intestine.

Introduction Most non-small-cell lung cancer (NSCLC) patients who exhibit good clinical responses to EGFR-tyrosine kinase inhibitors (EGFR-TKIs) will inevitably develop disease progression. Herein, through next-generation sequencing (NGS), we aimed to investigate three new disease-progression modes of NSCLC patients after EGFR-TKI treatment. Materials and Methods Patients with sensitive EGFR-mutations who acquired resistance to EGFR-TKIs and whose tissues were subjected to post-progression NGS were enrolled. The clinical characteristics, progression-free survival (PFS), genomic alterations and expression of EGFR-mutations among the three disease-progression modes were retrospectively analyzed. Results and Conclusion The sites of disease progression were as follows: primary foci in 19.5% (8/41) (Mode 1), metastatic foci in 31.7% (13/41) (Mode 2), and both primary and metastatic foci in 48.8% (20/41) (Mode 3). The median PFS in Mode 1 was 6 months (95% CI 1–8), which was significantly shorter than the 11 months (95% CI 8–14) in Mode 2 and the 10 months (95% CI 3–16) in Mode 3 (p = 0.0084). The expression of Del19 was significantly different among the three modes (p = 0.02). The numbers and species of mutant genes in Mode 3 were obviously greater than those in Modes 1 and 2, and no gene amplifications were observed in Mode 2. Mutations in the TP53 gene were the most frequent genetic alteration found in our study, and these accounted for 48.8% (20/41) of all alterations. TP53 mutations in Mode 1 were mainly in exons 6 and 8, while in Mode 2 and Mode 3, all mutations were located from exon 4 to exon 8. A significant benefit in PFS was observed in the metastatic foci progression mode and in the dual primary and metastatic foci progression mode rather than in the primary foci progression mode, which had significant value in the design of therapeutic strategies.