Background: Although the patients suffering from metastatic breast cancer (MBC) have experienced considerably wide survival ranges, reliable biomarkers for its prognosis have remained unidentified. This study attempted to profile the serum cytokine candidates that could potentially be used to predict the prognosis of MBC patients. Methods: By using the ProcartaPlex Multiplex Immunoassay, a total of 45 cytokines were assayed in sera from 18 MBC patients before any treatment. Results: Thirty-three types of cytokines were cross-detectable in more than 50% of the patients and thus were acceptable for a further analysis. Univariate analysis uncovered 10 sera cytokines significantly associated with MBC patient death. Among them, interleukin 8 (IL-8), fibroblast growth factor (FGF2), hepatocyte growth factor (HGF) and VEGF-A were reported, and 6 cytokines which included NGF-β, IFN-γ, IP-10, IL-18, MCP-1 and MIP-1β were identified for the first time as MBC death-correlated serum factors. Conclusions: Identification of these cytokines would facilitate developing novel serum biomarkers for MBC, favoring multiplexed serum cytokine profiling a promising method in predicting the outcome of MBC patients.

Neuronal PAS domain protein 3 (NPAS3) and VGF (VGF Nerve Growth Factor (NGF) Inducible) are important for neurogenesis and psychiatric disorders. Previously, we have demonstrated that NPAS3 regulates VGF at the transcriptional level. In this study, VGF (non-acronymic) was found regulated by NPAS3 in neuronal stem cells. However, the underlying mechanism of this regulation remains unclear. The aim of this study was to explore the correlation of NPAS3 and VGF, and their roles in neural cell proliferation, in the context of psychiatric illnesses. First, we focused on the structure of NPAS3, to identify the functional domain of NPAS3. Truncated NPAS3 lacking transactivation domain was also found to activate VGF, which suggested that not only transactivation domain but other structural motifs were also involved in the regulation. Second, Mutated enhancer box (E-box) of VGF promoter showed a significant response to this basic helix-loop-helix (bHLH) transcription factor, which suggested an indirect regulatory mechanism for controlling VGF expression by NPAS3. κB site within VGF promoter was identified for VGF activation induced by NPAS3, apart from direct binding to E-box. Furthermore, ectopically expressed NPAS3 in PC12 cells produced parallel responses for nuclear factor kappa-light-chain-enhancer of activated B cells [NF-κB (P65)] expression, which specifies that NPAS3 regulates VGF through the NF-κB signaling pathway. Over-expression of NPAS3 also enhances the cell proliferation, which can be blocked by knockdown of VGF. Finally, NPAS3 was found to influence proliferation of neural cells through VGF. Therefore, downstream signaling pathways that are responsible for NPAS3-VGF induced proliferation via glutamate receptors were explored. Combining this work and published literature, a potential network composed by NPAS3, NF-κB, Brain-Derived Neurotrophic Factor (BDNF), NGF and VGF, was proposed. This network collectively detailed how NPAS3 connects with VGF and intersected neural cell proliferation, synaptic activity and psychiatric disorders.

Schizophrenia and other major mental illnesses result from a complex interplay of genetic and environmental factors. We previously identified a mutation in NPAS3 that results in a valine to isoleucine (V304I) amino acid substitution segregating with schizophrenia in a small family. The amino acid change occurs in a potentially critical region for protein function. Furthermore, the same amino acid substitution in proteins related to familial Alzheimer's disease and transthyretin amyloidosis has been associated with protein aggregation. In this study, we demonstrate that NPAS3 is prone to aggregation, and that the V304I mutation in NPAS3 increases this propensity in both bacterial and mammalian expression systems. We also show that NPAS3-V304I reduces soluble endogenous NPAS3, and increases insoluble endogenous NPAS3 and leads to alteration of transcriptional activity. These results suggest that protein aggregation, potentially leading to cell dysfunction via a loss of protein function through sequestration, may contribute to the pathogenesis of schizophrenia and other forms of mental illness. Further exploration of the mechanisms leading to abnormal protein quality control could lead to new therapeutic targets.

OBJECTIVE:To observe the expression of nuclear protein 1 (Nupr1), a stress-related nuclear protein, in a panel of human hepatocellular carcinoma cell lines, and its effects on the proliferation and migration of hepatocellular carcinoma cells by RNA interference-mediated knockdown.METHODS:Real-time quantitative PCR was employed to detect Nupr1 mRNA levels in hepatocellular carcinoma cell lines, including BEL-7402, QSG-7703, SMMC-7721 and HepG2. After Nupr1 expression was knocked down by RNA interference, cell proliferation was monitored by MTT assay and colony formation in plate. Cell cycle was analyzed by flow cytometry and cell migration was assayed by TranswellTM assay.RESULTS:Higher expression of Nupr1 was detected in HepG2 as compared with the other cell lines. Nupr1 expression in HepG2 cells were efficiently knocked down by two short hairpin RNAs (shRNAs), with inhibitory rates being 72.25% and 84.25%, respectively. HepG2 cells with Nupr1 knockdown displayed lower rate of proliferation, G1 arrest, and significantly decreased abilities of cell migration and colony formation. Western blotting showed that Nupr1 knockdown increased the expressions of p21 and p27, two negative regulators of cell cycle.CONCLUSION:Knockdown of Nupr1 inhibited the proliferation and migration of HepG2 hepatocellular carcinoma cells.

OBJECTIVES:Ubiquitin carboxy-terminal hydrolase-L1 (UCH-L1) has been established as a potential biomarker of neuronal damage. There is not much information about the effects of white matter lesions (WMLs) on serum and urine UCH-L1 levels in white matter disease patients. This study was aimed to assess whether serum or urine UCH-L1 levels are a reliable marker of brain damage in patients with WMLs. DESIGN AND METHODS:Serum and urine levels of UCH-L1 were assessed in 125 patients with dizziness, hypertension, type 2 diabetes mellitus, or dyslipidemia. Of these 125 patient cases, 41 showed periventricular WMLs (P-WMLs), 46 showed subcortical WMLs (S-WMLs), and 38 displayed no well-defined WMLs (controls). RESULTS:Serum UCH-L1 levels were significantly different between the WML group and controls (p<0.05). Further subgroup analysis proved that serum UCH-L1 levels in participants with S-WMLs were significantly increased when compared with controls (p<0.001), but there was no significant differences between controls and patients with P-WMLs (p>0.05). However, urine levels of UCH-L1 were similar between these three groups (p>0.05). In addition, multivariate analysis showed that increased serum UCH-L1 levels were independently associated with the severity of WMLs using Fazekas scale (β=0.432, p<0.001). CONCLUSIONS:These findings suggest that serum UCH-L1 levels may serve as a novel biomarker for neuronal damage from WMLs, especially S-WMLs.

The functional study of the edible mycoprotein is important for elucidating the efficacy mechanism of fungi. There are more broad prospects on the field of antitumor mycoprotein. The expression map of Lentinula edodes C91-3 was sequenced and analyzed on the previous work. A novel antitumor mycoprotein-Latcripin-1 (LP1) was found at the first time. In order to clarify the mechanism of its inducing programmed cell death in lung cancer cells, protein LP1 PI3K (phosphatidylinositol 3-kinase) functional domain peptide was obtained by the Rosetta gami prokaryotic expression and purification system. Western blot was used to identify the functional domain and the autophagosomal marker-microtubule associated protein light chain 3. Transmission electron microscopy technology was used to analysed the ultrastructure of the treated cell. Flow cytometry and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) methods were used to detect the biological functions of domain PI3K to lung cancer cell. This work demonstrates that the functional domain PI3K of LP1 maybe induces lung cancer A549 to programmed cell death through autophagy pathway. The functional domain PI3K in L. edodes C91-3 should be ascribed to class III PI3K. Though further study is needed, it will shed new light into developing new anti-tumor polypeptide drugs.

Objective: Deficits in adult and embryonic neurogenesis have been linked with neurological and psychiatric disorders, so it is important to understand the molecular mechanisms underlying this process. SOX11 is a transcription factor known to play a critical role in the regulation of the neuronal and glial differentiation stage of neurogenesis, so we hypothesised that the identification of its target genes would reveal underlying biological processes relevant to disease.Methods: SOX11 protein was over-expressed in HEK293 cells and transcriptional changes assessed by microarray analysis. Selected candidate genes were further tested for SOX11 activation in quantitative reverse transcriptase PCR studies of HEK293 cells and Western analysis of SH-SY5Y cells.Results: Regulated genes included a previously established SOX11 target, known markers of neurogenesis, as well as several genes implicated in neuropsychiatric disorders. Immunofluorescence localised several of the genes within the proliferative subgranular zone of the hippocampus. We observed multiple histone and zinc finger genes regulated by SOX11, many of which were located in two clusters on chromosomes 6 and 19. The chromosome 6 cluster lies within a region of the genome showing the strongest genetic association with schizophrenia.Conclusion: SOX11 appears to regulate a complex programme of chromatin remodelling and downstream gene expression changes to achieve a mature neuronal phenotype. SOX11 target genes are shown to be involved in neurodevelopmental processes important in health and, potentially, disease.

The XVII World Congress of Psychiatric Genetics, sponsored by The International Society of Psychiatric Genetics (ISPG) took place in San Diego, California from 4 to 8 November 2009. Approximately 550 participants gathered to discuss the latest molecular genetic findings relevant to serious mental illness, including schizophrenia, mood disorders, substance abuse, autism, and attention deficit disorder. Recent advances in the field were discussed, including the genome-wide association studies results, copy number variation (CNV) in the genome, genomic imaging, and large multicenter collaborations. The following report, written by junior travel awardees who were assigned sessions as rapporteurs represents some of the areas covered in oral presentation during the conference, and reports on some of the notable major new findings described at this 2009 World Congress of Psychiatric Genetics.