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It is attractive but remains challenging to develop nanomaterials-based methods to realize the colorimetric detection of ultratrace biological analytes. In this work, an HAuCl 4 -H 2 O 2 system was developed for the visual detection of intracellular hydrogen peroxide (H 2 O 2 ) based on the Au-Pt/SiO 2 sensing platform. Owing to their excellent intrinsic peroxidase-like activity, Au-Pt/SiO 2 nanospheres were introduced to catalyze the decomposition of H 2 O 2 and then directly reduce HAuCl 4 into gold nanoparticles (Au NPs) for the naked-eye detection. The produced Au NPs behaved color variations from light purple to wine red as H 2 O 2 concentration increased. Under the optimal conditions, this strategy showed excellent sensitivity for H 2 O 2 with a detection limit of 0.1 nM by naked-eye readout and 0.1 pM by UV–vis spectrometer (S/N = 3), as well as acceptable selectivity for H 2 O 2 , which could meet the demand for the detection of H 2 O 2 in biological systems. The method was further validated by the determination of H 2 O 2 in HeLa cells with low cellular cytotoxicity. Thus, this strategy would provide simple and sensitive detection of H 2 O 2 in biological systems, revealing the potential application in clinical, biological and environmental researches.

Summary Currently, patients receiving cancer treatments routinely suffer from distressing toxic effects, most originating from premature drug leakage, poor biocompatibility, and off-targeting. For tackling this challenge, we construct an intracellular Ca2+ cascade for tumor therapy via photothermal activation of TRPV1 channels. The nanoplatform creates an artificial calcium overloading stress in specific tumor cells, which is responsible for efficient cell death. Notably, this efficient treatment is activated by mild acidity and TRPV1 channels simultaneously, which contributes to precise tumor therapy and is not limited to hypoxic tumor. In addition, Ca2+ possesses inherent unique biological effect and normal cells are more tolerant of the undesirable destructive influence than tumor cells. The Ca2+ overload leads to cell death due to mitochondrial dysfunction (upregulation of Caspase-3, cytochrome c, and downregulation of Bcl-2 and ATP), and in vivo, the released photothermal CuS nanoparticles allow an enhanced 3D photoacoustic imaging and provide instant diagnosis., Graphical Abstract, Highlights • The Ca2+ cascade is selectively constructed in vivo without being limited to hypoxic tumor • Ca2+ overload leads to mitochondrial dysfunction and cancer death • The released CuS NPs provide an enhanced 3D photoacoustic imaging • Ca2+-interference therapy avoids the obstacles of traditional treatment, Medical Imaging; Nanoparticles; Cancer

Currently, patients receiving cancer treatments routinely suffer from distressing toxic effects, most originating from premature drug leakage, poor biocompatibility and off-targeting. For tackling this challenge, we construct an intracellular Ca2+ cascade for tumor therapy via photothermal activation of TRPV1 channels. The nanoplatform creates an artificial calcium overloading stress in specific tumor cells, which is responsible for efficient cell death. Notably, this efficient treatment is activated by mild acidity and TRPV1 channels simultaneously, which contributes to precise tumor therapy and is not limited to hypoxic tumor. In addition, Ca2+ possesses inherent unique biological effect and normal cells are more tolerant of the undesirable destructive influence of NPs than tumor cells. The Ca2+ overload leads to cell death due to mitochondrial dysfunction (upregulation of Caspase-3, cytochrome c, and downregulation of Bcl-2 and ATP) and in vivo, the release photothermal CuS NPs allows an enhanced 3D photoacoustic (PA) imaging and provide instant diagnosis.

Breast cancer is the leading cause of cancer-related death among women, with a more 20% 5-year survival rate after metastases. It is therefore critical to improve early diagnosis in order to improve disease prognosis. This study investigates cytokine profiles of breast cancer serum with the aim of identifying biomarkers for early diagnosis. A solid-phase antibody array was used for screening 274 biomarkers in serum from breast cancer patients. ELISA assay was carried out to identify biomarkers with differential expression. The serum levels of IL-8, MIP-1 alpha, MIP-1 beta, MMP-8, Resistin, FLRG, and BCAM were significantly higher in breast cancer patients, but LAP and TSH-β levels were lower. ELISA assay results confirmed those of the antibody array. Our results suggest that these cytokines, screened by antibody array, might serve as novel inflammatory markers in breast cancer patients. Whether these biomarkers are specific for breast cancer and can help to improve diagnoses and prognoses of breast cancer needs further investigation.

// Linhai Li 1, * , Lidan Chen 1, * , Jun Li 2, * , Weiyun Zhang 1 , Yang Liao 1 , Jianyun Chen 1 , Zhaohui Sun 1 1 Department of Laboratory Medicine, Guangzhou General Hospital of Guangzhou Military Command of PLA, Guangzhou, Guangdong 510010, P.R. China 2 Department of Information, No.4 Hospital of PLA, XiNing, Qinghai 810007, P.R. China * These authors have contributed equally to this work Correspondence to: Linhai Li, e-mail: mature303@126.com Keywords: mitochondrial DNA, mutation, high-throughput sequencing, breast cancer Received: January 01, 2016 Accepted: April 02, 2016 Published: April 21, 2016 ABSTRACT The presented study performed an mtDNA genome-wide association analysis to screen the peripheral blood of breast cancer patients for high-risk germline mutations. Unlike previous studies, which have used breast tissue in analyzing somatic mutations, we looked for germline mutations in our study, since they are better predictors of breast cancer in high-risk groups, facilitate early, non-invasive diagnoses of breast cancer and may provide a broader spectrum of therapeutic options. The data comprised 22 samples of healthy group and 83 samples from breast cancer patients. The sequencing data showed 170 mtDNA mutations in the healthy group and 393 mtDNA mutations in the disease group. Of these, 283 mtDNA mutations (88 in the healthy group and 232 in the disease group) had never been reported in the literature. Moreover, correlation analysis indicated there was a significant difference in 32 mtDNA mutations. According to our relative risk analysis of these 32 mtDNA mutations, 27 of the total had odds ratio values (ORs) of less than 1, meaning that these mutations have a potentially protective role to play in breast cancer. The remaining 5 mtDNA mutations, RNR2-2463 indelA, COX1-6296 C>A, COX1-6298 indelT, ATP6-8860 A>G, and ND5-13327 indelA, whose ORs were 8.050, 4.464, 4.464, 5.254 and 4.853, respectively, were regarded as risk factors of increased breast cancer. The five mutations identified here may serve as novel indicators of breast cancer and may have future therapeutic applications. In addition, the use of peripheral blood samples was procedurally simple and could be applied as a non-invasive diagnostic technique.

Photodynamic therapy suffers from poor tumor selectivity and poor therapeutical efficacy. In this paper, an amphiphilic chimeric peptide is fabricated to realize sequential acidity-responsive tumor-targeted transport of photosensitizer and in situ photodynamic therapy in nuclei. In vitro studies demonstrate that the acidic tumor microenvironment successfully sheds the mask of cationic nuclear localization sequence (NLS) of the negatively charged chimeric peptide. This charge reversal remarkably accelerates cellular uptake of chimeric peptide in tumor cells and maximizes the photodynamic therapeutical efficacy in nuclei. Most importantly, direct disguise of the biofunctional NLS sequence decreases the complexity and increases the performance of the chimeric peptide further by achieving long blood retention time, specific tumor accumulation, minimal side effects, and efficient antitumor therapy in vivo.

The miniaturization of gold nanorods exhibits a bright prospect for intravital photoacoustic imaging (PAI) and the hollow structure possesses a better plasmonic property. Herein, miniature hollow gold nanorods (M-AuHNRs) (≈46 nm in length) possessing strong plasmonic absorbance in the second near-infrared (NIR-II) window (1000-1350 nm) are developed, which are considered as the most suitable range for the intravital PAI. The as-prepared M-AuHNRs exhibit 3.5 times stronger photoacoustic signal intensity than the large hollow Au nanorods (≈105 nm in length) at 0.2 optical density under 1064 nm laser irradiation. The in vivo biodistribution measurement shows that the accumulation in tumor of miniature nanorods is twofold as high as that of the large counterpart. After modifying with a tumor-targeting molecule and fluorochrome, in living tumor-bearing mice, the M-AuHNRs group gives a high fluorescence intensity in tumors, which is 3.6-fold that of the large ones with the same functionalization. Moreover, in the intravital PAI of living tumor-bearing mice, the M-AuHNRs generate longer-lasting and stronger photoacoustic signal than the large counterpart in the NIR-II window. Overall, this study presents the fabrication of M-AuHNRs as a promising contrast agent for intravital PAI.

Photodynamic therapy (PDT) brings excellent treatment outcome while also causing poor tumor microenvironment and prognosis due to the uncontrolled oxygen consumption. To solve this issue, a novel PDT strategy, oxygenated PDT (maintain the tumor oxygenation before and after PDT) was carried out by a tumor and apoptosis responsive photoactivity conversion nanocomposite (MPPa-DP). Under physiological conditions, this nanocomposite has a low photoactivity. While at H

Glutamate Ionotropic Receptor Kainate Type Subunit 3 (GRIK3) is an important excitatory neurotransmitter receptor that plays a significant role in various neurodegenerative diseases. However, the biological functions of GRIK3 in malignancies are largely unknown because of limited related studies. Here, we primarily reported that the expression of GRIK3 was higher in breast cancer tissues than in adjacent noncancerous tissues. GRIK3 expression was also positively correlated with the prognosis of patients with breast cancer. GRIK3 promoted the proliferation and migration abilities of breast cancer cells and enhanced the growth of orthotopically implanted tumors. Mechanically, GRIK3 influenced a range of signaling pathways and key signal transducers, including two epithelial‐mesenchymal transition regulators, SPDEF and CDH1. Heterogenous expression of SPDEF and CDH1 counteracted the migration and invasion abilities, respectively, of breast cancer cells induced by GRIK3. Moreover, overexpression of GRIK3 increased the expression of mesenchymal markers and decreased the expression of epithelial markers, resulting in the translocation of β‐catenin into the nucleus and the increased β‐catenin transcriptional activity. In conclusion, the present study reported a novel oncogenic role of GRIK3. Meanwhile, GRIK3, as a membrane receptor, may also serve as a potential therapeutic target for the treatment of breast cancer.

Hepatitis B e-antigen (HBeAg) is the secretory form of the nucleocapsid of the hepatitis B virus (HBV), which is a marker of viral replication. In this study, a novel signal amplification system (SAS) based on the lateral flow immunoassay (LFIA) was used for rapid detection of HBeAg in blood samples from patients or blood donors. In this assay, the detection antibody was conjugated with gold nanoparticles (GNPs), and the capture antibody was labeled with biotin. The presence of targeting antigen HBeAg in blood sample would act as a bridge with biotinylated captured antibody and GNP-conjugated detection antibody to form the dendritic nanoparticle complex. The dendritic complexes in the sample solution were migrated and immobilized on the testing line of strip coated with antibiotin antibodies. Signal intensity was massively amplified by the SAS, which was positively correlated with the concentration of targeting antigen in the blood sample and was assessed by eyes or strip scanner. The SAS worked only when targeting antigens were present in the sample. By using this SAS-LFIA, we were able to detect a very low concentration of HBeAg (9 ng/mL), which was 27-fold sensitive than that by conventional LFIA (cLFIA). A number of 420 blood samples were detested by this novel SAS-LFIA, the results were in accordance with those of enzyme-linked immunosorbent assay (ELISA) completely, while the cLFIA missed an HBeAg-positive sample. In conclusion, the novel SAS has high specificity and sensitivity, which can be used to replace the conventional rapid test and ELISA in clinical diagnosis.

Few prognostic indicators with differential expression have been reported among the differing ER statuses. We aimed to screen important breast cancer prognostic genes related to ER status and to construct an efficient prognostic prediction system. mRNA expression profiles were downloaded from TCGA and GSE70947 dataset. Two hundred seventy-one overlapping differentially expressed genes (DEGs) between the ER- and ER+ breast cancer samples were identified. Among the 271 DEGs, 109 prognostically relevant mRNAs were screened. mRNAs such as RASEF, ITM2C, CPEB2, ESR1, ANXA9, and VASN correlated strongly with breast cancer prognosis. Three modules, which contained 28, 9 and 8 enriched DEGs, were obtained from the network, and the DEGs in these modules were enriched in response to hormone stimulus, epithelial cell development, and host cell entry. Using bayes discriminant analysis, 48 signature genes were screened. We constructed a prognostic prediction system using the 48 signature genes and validated this system as relatively accurate and reliable. The DEGs might be closely associated with the prognosis in patients with breast cancer. We validated the effectiveness of our prognostic prediction system by GEO database. Therefore, this system might be a useful tool for preliminary screening and validation of potential prognosis indicators for ER+ breast cancer derived from mechanistic research.

Figure S3. Distribution of mRNA expression density. Solid and dashed lines indicate the density distribution curve before and after the removal of mRNA with low expression levels, respectively. (PDF 7 kb)

Figure S2. Distribution of SOSTDC1 methylation levels in the control and luminal breast cancer tissues. Red and blue points indicate methylation levels in the control and cancer samples, respectively. Black lines indicate the mean methylation levels of the corresponding samples. (PDF 19 kb)

Figure S5. Kaplanâ Meier survival analysis based on risk score model system (a) and Luminal subtypes using the Metabric cohort (b). (a) The black and red lines indicates the low-risk group and the high-risk group; (b) The black and red lines indicates the Luminal A and Luminal B breast cancer tissues. (PDF 29 kb)

Figure S1. Box plot showing the methylation levels of 14 genes in the control and luminal breast cancer tissues. Blue and orange boxes indicate methylation levels in the control and cancer samples, respectively. (PDF 20 kb)

Background: With the development of more rapid and sensitive detection methods based on PCR techniques, the contributions of respiratory viral infections to community-acquired pneumonia (CAP) in adult patients are being more and more recognized. Yet, up to now, there has been a lack of synthetic data that clearly demonstrates the incidence of respiratory viral infections in adult patients with CAP. Objectives: We intended to demonstrate the incidence of respiratory viral infections detected by PCR and real-time PCR in adult patients with CAP. Methods: We searched PubMed and Embase for studies providing the incidence of respiratory viral infections in adult patients with CAP. We investigated potential sources of heterogeneity by a univariant metaregression analysis and calculated the combined incidence of viral infections, viral infections mixed with other pathogens and individual respiratory virus species. Results: We eventually identified 23 eligible reports with a total number of 6,404 patients. Incidences ranged from 8.6 to 56.2% for overall respiratory viral infections. We noted significant heterogeneity in incidence estimates for the incidence of viral infections (Cochran's χ2 = 269.9, p < 0.0001, I2 = 91.8%). The combined incidence of viral infections was 22.4% (95% CI = 19.0-25.7). Incidences of viral coinfections with other pathogens ranged from 3 to 28%. A high level of heterogeneity was identified as well during the estimates for incidences of coinfections (χ2 = 200.9, p < 0.0001, I2 = 91.5%). The combined incidence of viral coinfections with other pathogens was 12.4% (95% CI = 9.7-15.0). Our heterogeneity analyses suggested that a lower respiratory tract sample was associated with higher overall viral incidence. Moreover, the influenza virus, rhinovirus and coronavirus were the 3 most frequently detected viral pathogens in adult patients with CAP according to our study. Conclusions: Respiratory viruses are probably crucial pathogens of adult patients with CAP, with the influenza virus being the most frequent viral pathogen identified. More than half of the viral infections are characterized as mixed infections with other pathogens.

Background Robust and precise molecular prognostic predictors for luminal breast cancer are required. This study aimed to identify key methylation sites in luminal breast cancer, as well as precise molecular tools for predicting prognosis. Methods We compared methylation levels of normal and luminal breast cancer samples from The Cancer Genome Atlas dataset. The relationships among differentially methylated sites, corresponding mRNA expression levels and prognosis were further analysed. Differentially expressed genes in normal and cancerous samples were analysed, followed by the identification of prognostic signature genes. Samples were divided into low- and high-risk groups based on the signature genes. Prognoses of low- and high-risk groups were compared. The Gene Expression Omnibus dataset were used to validate signature genes for prognosis prediction. Prognosis of low- and high-risk groups in Luminal A and Luminal B samples from the TCGA and the Metabric cohort dataset were analyzed. We also analysed the correlation between clinical features of low- and high- risk groups as well as their differences in gene expression. Results Fourteen methylation sites were considered to be related to luminal breast cancer prognosis because their methylation levels, mRNA expression and prognoses were closely related to each other. The methylation level of SOSTDC1 was used to divide samples into hypo- and hyper-methylation groups. We also identified an mRNA signature, comprising eight transcripts, ESCO2, PACSIN1, CDCA2, PIGR, PTN, RGMA, KLK4 and CENPA, which was used to divide samples into low- and high-risk groups. The low-risk group showed significantly better prognosis than the high-risk group. A correlation analysis revealed that the risk score was an independent prognostic factor. Low- and high- risk groups significantly correlated with the survival ratio in Luminal A samples, but not in Luminal B samples on the basis of the TCGA and the Metabric cohort dataset. Further functional annotation demonstrated that the differentially expressed genes were mainly involved in cell cycle and cancer progression. Conclusions We identified several key methylation sites and an mRNA signature for predicting luminal breast cancer prognosis. The signature exhibited effective and precise prediction of prognosis and may serve as a prognostic and diagnostic marker for luminal breast cancer. Electronic supplementary material The online version of this article (10.1186/s12885-018-4314-9) contains supplementary material, which is available to authorized users.

Estrogen receptor-positive (ER+) and ER-negative (ER-) subtypes of breast cancer have distinct clinical outcomes because they respond differentially to endocrine therapies. We aimed to comprehensively analyze differentially expressed microRNA (miRNAs), long non-coding RNAs (lncRNAs) and mRNAs in different ER subtypes as well as to identify prognosis-related RNAs. The expression levels of miRNAs, lncRNAs, and mRNAs between breast cancer and normal samples were compared using data from The Cancer Genome Atlas database. Differentially expressed miRNAs, lncRNAs and mRNAs between ER+ and ER- samples were also screened. An ER subtype-related miRNA-lncRNA-mRNA network was constructed. lncRNAs and mRNAs in this network were further subjected to an analysis of their associations with patient prognosis. Sets of differentially expressed miRNAs, lncRNAs, and miRNAs between breast cancer and normal samples were identified among which 14 miRNAs, 78 lncRNAs, and 475 mRNAs were differentially expressed between ER subtypes. Relationships between these RNAs were analyzed. The resultant ER subtype-related miRNA-lncRNA-mRNA network consisted of 14 nodes, among which LINC0092 and chromosome 2 open reading frame 71 (C2orf71) were correlated with better prognosis of breast cancer. LINC0092 was co-expressed with SFRP1 and RGMA and regulated by hsa-miR-449a and hsa-miR-452-5p. C2orf71 was co-expressed with LINC00511 and regulated by hsa-miR-184. Cross-talk among differentially expressed miRNAs, lncRNAs, and miRNAs may be an important feature in ER+ and ER- subtypes of breast cancer. LINC0092 and C2orf71, two of these cross-talking RNAs, may serve as novel prognostic predictor of breast cancer because of their close associations with prognosis.

Natural killer (NK) cells are critical components in innate immune response to viral infection. Killer cell immunoglobulin-like receptors (KIRs) are involved in regulating the balance of activation or inhibitory function of NK cells. However, the association of KIRs with the spontaneous clearance of hepatitis C virus (HCV) remains unclear in the Chinese population.A total of 407 HCV-seropositive voluntary blood donors were recruited, including 203 with spontaneous viral clearance and 204 with chronic infection. The presence of KIR genes was detected individually by polymerase chain reaction with sequence-specific primers. Data of HLA and interleukin-28B (IL28B) genotypes were extracted from our previous study.Our results showed that KIR2DL2, 2DS2, 2DL2/2DL3, and 2DL5A-/2DL5B+ were more frequent in subjects with HCV clearance than those with chronic infection (odds ratio [OR], 1.640, p = 0.034; OR, 1.664, p = 0.032; OR, 1.636, p = 0.040; and OR, 2.601, p = 0.012, respectively). Multivariate logistic regression analysis showed that KIR2DL5A-/2DL5B+ associated with HCV clearance (OR, 2.448, p = 0.027), independent of sex, IL28B, and other KIRs. In contrast, KIR2DL3/2DL3 (OR, 0.610, p = 0.034) as well as 2DL3/2DL3+HLA-C1 or C1C1 (OR, 0.580, p = 0.017; and OR, 0.639, p = 0.025, respectively) was found associated with chronic HCV infection. The presence of the homozygous KIR2DL3 with or without its HLA ligand increased the OR of developing chronic HCV infection in the context of IL28B.In this study we identified KIR2DL5A-/2DL5B+ associated with HCV spontaneous clearance, while KIR2DL3/2DL3, 2DL3/2DL3+HLA-C1, or C1C1 associated with chronic infection. Our study highlighted the fact that the roles of KIR and KIR-HLA contributed to the control of HCV infection by innate immune responses.

Photodynamic therapy (PDT) holds great promise in tumor treatment. Nevertheless, it remains highly desirable to develop easy-to-fabricated PDT systems with improved tumor accumulation/internalization and timely therapeutic feedback. Here, we report a tumor-acidity-responsive chimeric peptide for enhanced PDT and noninvasive real-time apoptosis imaging. Both in vitro and in vivo studies revealed that a tumor mildly acidic microenvironment could trigger rapid protonation of carboxylate anions in chimeric peptide, which led to increased ζ potential, improved hydrophobicity, controlled size enlargement, and precise morphology switching from sphere to spherocylinder shape of the chimeric peptide. All of these factors realized superfast accumulation and prolonged retention in the tumor region, selective cellular internalization, and enhanced PDT against the tumor. Meanwhile, this chimeric peptide could further generate reactive oxygen species and initiate cell apoptosis during PDT. The subsequent formation of caspase-3 enzyme hydrolyzed the chimeric peptide, achieving a high signal/noise ratio and timely fluorescence feedback. Importantly, direct utilization of the acidity responsiveness of a biofunctional Asp-Glu-Val-Asp-Gly (DEVDG, caspase-3 enzyme substrate) peptide sequence dramatically simplified the preparation and increased the performance of the chimeric peptide furthest.

Geometrical shape of nanoparticles plays an important role in cellular internalization. However, the applicability in tumor selective therapeutics is still scarcely reported. In this article, we designed a tumor extracellular acidity-responsive chimeric peptide with geometrical shape switch for enhanced tumor internalization and photodynamic therapy. This chimeric peptide could self-assemble into spherical nanoparticles at physiological condition. While at tumor extracellular acidic microenvironment, chimeric peptide underwent detachment of acidity-sensitive 2,3-dimethylmaleic anhydride groups. The subsequent recovery of ionic complementarity between chimeric peptides resulted in formation of rod-like nanoparticles. Both in vitro and in vivo studies demonstrated that this acidity-triggered geometrical shape switch endowed chimeric peptide with accelerated internalization in tumor cells, prolonged accumulation in tumor tissue, enhanced photodynamic therapy, and minimal side effects. Our results suggested that fusing tumor microenvironment with geometrical shape switch should be a promising strategy for targeted drug delivery.

In 2014, an outbreak of dengue virus (DENV) infection led to 45 171 clinical cases diagnosed in Guangdong province, Southern China. However, the potential risk of blood donors asymptomatically infected with DENV has not been evaluated . In the current study we detected anti-DENV IgG antibody and RNA in volunteer Chinese blood donors. We found that anti-DENV IgG antibody was positively detected in 3.4% (51/1500) and two donors were detected as being DENV RNA positive out of 3000 blood samples. We concluded that the presence of potential DENV in blood donors might be potential risk for blood safety. Therefore, screening for DENV infection should be considered in blood donations during a period of dengue outbreak in high epidemic area of China.

The aim of this study is to investigate protective effects of sesaminol on the human bronchial epithelial (BEAS-2B) cell line against oxidative damage of cigarette smoke extract (CSE). BEAS-2B cells were pre-incubated with sesaminol for 12 h and then treated with various concentrations of CSE for 24 h. After that proliferation ability, levels of reactive oxygen species (ROS) and lactate dehydrogenase (LDH), cell apoptosis, activities of catalase (CAT) and superoxide dismutase (SOD), and mRNA levels of IL-8 and IL-6 were measured. The results showed that sesaminol significantly improved BEAS-2B cell viability, reduced the production of ROS and LDH of cells, inhibited cell apoptosis and increased CAT and SOD activities in CSE-treated cells. Sesaminol also inhibited the expression of IL-8 and IL-6 mRNA following CSE exposure. In conclusion, sesaminol may protect BEAS-2B cells against CSE-induced oxidative damage.

SRY-related high-mobility-group box 9 (SOX9) is a member of the SOX family of transcription factors. Accumulating evidence has shown that SOX9 plays a significant role in various malignancies. However, the role of SOX9 in nasopharyngeal carcinoma (NPC) remains unknown. In the present study, up-regulation of SOX9 was observed in both NPC tissues and different NPC cells. Overexpression of SOX9 promoted NPC cell proliferation, migration and invasion. Conversely, knock down of SOX9 inhibited NPC proliferation, colony formation, migration and invasion. Mechanistically, SOX9 bound directly to the promoter region of BMP2 and increased BMP2 expression. In addition, overexpression of SOX9 activated the mTOR pathway partly through BMP2. Collectively, these results identify a novel role for SOX9 as a potential therapeutic marker for the prevention and treatment of NPC.

Photodynamic therapy (PDT) brings excellent treatment outcome while also causing poor tumor microenvironment and prognosis due to the uncontrolled oxygen consumption. To solve this issue, a novel PDT strategy, oxygenated PDT (maintain the tumor oxygenation before and after PDT) was carried out by a tumor and apoptosis responsive photoactivity conversion nanocomposite (MPPa-DP). Under physiological conditions, this nanocomposite has a low photoactivity. While at H2O2-rich tumor microenvironment, the nanocomposite could react with overexpressed H2O2 to produce O2 and release high photoactivity chimeric peptide PPa-DP for oxygenated tumor and PDT. Importantly, when the PDT mediates cell apoptosis, the photoactivity of PPa-DP be effectively quenched and the O2 consumption appeared retard, which avoided further consumption of residual O2 on apoptotic cells. In vitro and vivo studies revealed that this nanocomposite could efficiently change photoactivity, reasonable control O2 consumption and increase residual O2 content of tumor after PDT.

Mitochondrial DNA mutations have been identified in serveral types of cancer. In breast cancer, germline and somatic mitochondrial DNA (mtDNA) mutations have been identified. A number of mtDNA mutations in breast cancer have been identified in protein-coding regions (in protein-coding genes, such as ND2, COX3, ND4, ND5 and CytB). Mutations in these structure proteins cause impaired electron transport function and lead to electron leakage and increased reactive oxygen species (ROS) production, which in turn increases oxidative stress and oxidative damage to the mitochondria, as well as to cells. These data establish an association between mtDNA mutations and breast cancer; however, there is no reliable prediction of breast cancer predisposition or progression based on mtDNA mutation patterns thus far. In this study, we used high-throughput sequencing to detect mtDNA mutations in the blood of breast cancer patients. Some of these mutations may be used as potential markers for breast cancer diagnosis.

Edible vegetable oil usually produces high concentration triglyceride polymer (TGP) after highly oxidative deterioration, and the toxic effects of TGP in animals are still under debate, with no definitive evidence supporting the toxicity of TGP in animals. The aim of the present study was to evaluate the toxicity of TGP on murine immunocytes. Cultured macrophages were treated with different concentrations of TGP, and then the quantity of live cells, phagocytic activity and lactate dehydrogenase (LDH) release were determined. Additionally, apoptosis was observed by laser scanning confocal microscopy and flow cytometry. The intracellular reactive oxygen species (ROS) level was also determined by flow cytometry. The results showed that the viability and phagocytic activity of macrophages were significantly inhibited by TGP, and elevated LDH release was observed in TGP-treated macrophages. TGP also significantly increased the intracellular ROS level and thus resulted in apoptosis of macrophages. These results suggest that TGP has obvious toxicity against immune function. Practical applications: Triacylglycerols in edible vegetable oil may form polar oxidized TGP by oxidization during the processes of storage, cooking and refining, especially high-temperature frying. The present study has demonstrated that TGP may cause significant toxicity in macrophages and therefore it can be employed as a quality control index for consumer cooking oil.

Ethnopharmacological relevance Cordyceps sinensis has been used as a precious herbal medicine for thousands of years in China. Its polysaccharide fraction has been confirmed possessing immunomodulatory function and we have reported the acid polysaccharide fraction (APSF), from an anamorph of C. sinensis , has stimulating activity on macrophages. The mechanism still needs to be further elucidated. Materials and methods In order to investigate the effects of APSF on macrophage's phenotypes, Ana-1 mouse macrophages were polarized to M2 phenotype by culturing the cells with culture supernatant of H22 cells. M2 phenotype was determined by measuring the expression of TNF-α and checking cell surface markers mannose receptor (MR) and scavenger receptor (SR). After cultured with H22 supernatant for 72 h, the TNF-α level of Ana-1 cells was decreased while the SR and MR expressions were up-regulated, suggesting that Ana-1 cells were polarized towards M2 macrophages. Then the effects of APSF on M2 macrophages were investigated by measuring mRNA levels of TNF-α, inducible nitric oxide synthase (iNOS), IL-12 and IL-10. Nuclear NF-κB was detected by Western blotting. Results APSF treatment increased the expressions of TNF-α, IL-12 and iNOS, and reduced the expression of IL-10 of Ana-1 cells. Besides, the expressions of SR and MR were down-regulated by APSF. And the result of Western blotting showed NF-κB level was decreased in M2 macrophages and up-regulated after APSF treatment. Conclusions APSF may convert M2 macrophages to M1 phenotype by activating NF-κB pathway.

Cordyceps sinensis has been regarded as a precious tonic food and herbal medicine in China for thousands of years. The exopolysaccharide (EPS) from an anamorph of Cordyceps sinensis was found to have antitumor immunomodulatory activity. Mature dendritic cells play a role in initiating antitumor immunity, so we try to investigate the effects of EPS on the murine dendritic cell line DCS.Flow cytometry was used to assay the expression levels of cell surface molecules including major histocompatibility complex (MHC)-II, CD40, CD80, and CD86 of DCS cells and their ability to take up antigens. The ability of DCS cells to activate the proliferation of CTLL-2 T cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method. IL-12 and TNF-α levels were detected using ELISA. Western blotting was performed to estimate the levels of phosphorylated Janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), nuclear factor-κB (NF-κB) p65 and p105.EPS increased the expressions of MHC-II, CD40, CD80, and CD86 of DCS cells and up-regulated their ability to take up antigens. EPS also enhanced their ability to activate the proliferation of CTLL-2 T cells. IL-12 and TNF-α secreted from DCS cells were up-regulated after EPS treatment. Furthermore, EPS significantly caused the decline of p-JAK2 and p-STAT3, significantly increased levels of NF-κB p65 in the nucleus and decreased levels of NF-κB p105 in the cytoplasm.EPS may induce DCS cells to exhibit mature characteristics, and the mechanism involved is probably related to the inhibition of the JAK2/STAT3 signal pathway and promotion of the NF-κB signal pathway.

Dendritic cells are central players in initiating and regulating immune responses. Effective induction of cell-mediated immune responses strongly depends on stimulating T cells by dendritic cells through antigen-presenting complex and co-stimulatory molecules. In this study, dendritic cells were differentiated from human peripheral blood mononuclear cells by using rhGM-CSF and rhIL-4. Then the effects of exopolysaccharide (EPS) from an anamorph of Cordyceps sinensis on human DCs were investigated. The results of flow cytometry showed that surface HLA-DR and co-stimulatory molecules of DCs were increased and DC's phagocytosis of FITC-dextran were significantly decreased after EPS treatment. EPS-treated DCs also displayed enhanced T cell stimulatory capacity in mixed lymphocyte reaction (MLR). ELISA and RT-PCR assay showed EPS increased IL-12 level of DCs. RT-PCR assay also showed EPS decreased VEGF mRNA level of DCs. Furthermore, Western blotting demonstrated that phosphorylated STAT3 were decreased after EPS treatment, suggesting that EPS stimulated the maturation of DCs probably by inhibiting STAT3 activation.

Background and objective: Airway bacterial infections pose a significant challenge to the management of COPD, a disease mainly caused by cigarette smoking. However, the mechanisms of impaired airway mucosal innate immunity against bacteria in COPD remain unclear. We examined the effect of cigarette smoke on prostaglandin E2 (PGE2) and downstream epithelial host defence mechanisms including the antimicrobial substance human β-defensin-2 (hBD-2). Methods: Brushed bronchial epithelial cells were obtained from healthy smokers and individuals with COPD, and cultured under air–liquid interface conditions with or without exposure to whole cigarette smoke (WCS) or Moraxella catarrhalis (Mc) infection. Bacterial load, hBD-2 (a molecule known to kill Mc) and PGE2 were measured. Results: WCS decreased Mc-induced hBD-2 expression and increased Mc load on bronchial epithelial cells from healthy smokers and COPD patients. Moreover, WCS inhibited PGE2 induction following Mc. PGE2 was shown to increase hBD-2 production in bronchial epithelial cells from healthy smokers, but not from COPD patients. Conclusions: The results suggest that in well-differentiated human bronchial epithelial cells, WCS may impair host defence against Mc in part through inhibiting PGE2 production.

Cordyceps sinensis as a well-known traditional Chinese tonic has many therapeutic functions. In the present study, an acid polysaccharide (APS) was isolated from cultivated Cordyceps mycelia by ion-exchange and sizing chromatography. The protective capacity of APS against H(2)O(2)-induced oxidative damage in rat pheochromocytoma PC12 cells was investigated by measuring cell viability, lactate dehydrogenase (LDH) release, antioxidant enzyme activity, malondialdehyde (MDA) levels and intracellular accumulation of reactive oxygen species (ROS) and Ca(2+). The results demonstrated that pretreatment of PC12 cells with APS, prior to H(2)O(2) exposure, significantly increased the survival of cells and the activities of glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD), and reduced the levels of LDH and MDA. Intracellular accumulation of reactive oxygen species (ROS) and Ca(2+) were also inhibited by APS treatment. In conclusion, APS was found to increase the cellular antioxidant defence capacity, thereby protecting PC12 cells against oxidative stress.

As a freshwater pearl mussel, Hyriopsis cumingii is widely cultured in China to produce pearls. This study was made to explore the antitumor activity of water-soluble polysaccharide (WSP) from mantles of H. cumingii. Cell viability of human hepatocarcinoma HepG2 cells was estimated by MTT method. Cell cycle analysis was determined by flow cytometry. Apoptosis was observed under fluorescence microscopy and confirmed by DNA fragment assay. Reverse transcriptase-polymerized chain reaction (RT-PCR) and immunocytochemistry were carried to evaluate c-myc, bcl-2 and cyclinD1 gene expressions in HepG2 cells. Furthermore, intracellular reactive oxygen species (ROS) production was assessed by flow cytometry. After WSP treatment, the growth of HepG2 cells was inhibited and most of cells arrested in G0/G1 phage (p < .01); apoptotic changes in nucleus and cytoplasm were also observed in WSP-treated cells; percentage of apoptotic rate significantly increased in a dose-dependent manner (p < 0.001); DNA fragmentation was detected in treated HepG2 cells; The mRNA level and protein level of c-myc, bcl-2 and cyclinD1 were decreased in the treated HepG2 cells. ROS was significantly increased in a dose- and time-dependent manner as well. The results suggested that WSP has potent antitumor activity.

Two new triterpenoids, 3β-hydroxyoleana-11,13(18)-diene-28,30-dioic acid (1) and 3-oxooleana-11,13(18)-diene-28,30-dioic acid (2), one novel triterpenoid glycoside, 3β-O-(6'-O-methyl-β-D-glucuronopyranosyl)oleana-11,13(18)-dien-28-oic acid (3) along with six known compounds (4 - 9) were isolated from the stem bark of Aralia armata (Wall.) Seem. Their structures were elucidated through extensive spectroscopic methods. The herbicidal activities of these compounds against Bidens pilosa L., an invasive weed in P. R. China, were evaluated. Compounds 3, 5, and 6 exhibited more significant herbicidal activities on B. pilosa than the positive-control pendimethalin. Their possible use as herbicidal chemicals or model compounds deserved more attention. The effects of compounds 1 - 9 on Spodoptera litura cultured cell line Sl-1 cell proliferation and its morphology were also evaluated. The results indicated that compounds 1 - 5 affected Sl-1 cell proliferation. Compound 3 showed more obvious proliferation inhibition activities on Sl-1 cell than the positive-control rotenone. With regard to the effect on morphology, compound 2 significantly changed Sl-1 cell, resulting in cell blebbing and vacuole forming. Triterpenoids aremedicinally and agriculturally important, and cytotoxicity of the three new compounds 1 - 3 deserved further studies.

The effects of the immunomodulation and antitumor metastasis of the Chinese medicinal fungus water extract (FWE) were investigated. After B16 melanoma cells were implanted in C57BL/6 mice through their tail veins, the mice were orally administered by FWE at low dose (160 mg/kg/day) and high dose (400 mg/kg/day) for 14 days. The tumor metastatic foci on the lung surface were observed under dissecting microscope. Phagocytosis of peritoneal macrophages was detected through ingesting chicken red blood cells (CRBC) test. Lymphocyte proliferation and the natural killer (NK) cell activity were determined by MTT method and lactate-dehydrogenase (LDH)-release assay, respectively. The expressions of bcl-2 and p53 of livers and lungs were assayed by immunohistochemical method. The results showed that FWE could increase body weights of mice dose dependently. The lung metastasis inhibition rates (MIR) of B16 melanoma cells by low dose and high dose of FWE were 15.5% and 72.7%, respectively. FWE at the two doses enhanced significantly (p

There is increasing evidence that psychological stress and depression trigger changes in various biochemical parameters in animals and in human subjects. In order to study these effects, the impact of chronic mild stress (CMS) on rats, and of the subsequent administration of Banxia-houpu decoction and fluoxetine, were studied regarding their effects on the following biochemical parameters: 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) levels in various brain regions, natural killer (NK) cell and lymphokine-activated killer (LAK) cell activities in spleen, serum lipid profiles including total cholesterol (TC), high density lipoprotein cholesterol (HDLc), low density lipoprotein cholesterol (LDLc) and triglyceride (TG), liver superoxide dismutase (SOD) and nitric oxide synthase (NOS) activities, serum malondialdehyde (MDA), and interleukin-2 (IL-2) levels. The effects of drug administration on preference behavior for consumption of sucrose solution were also assessed. Rats subjected to CMS exhibited a reduction in sucrose intake, 5-HT, 5-HIAA, IL-2, TC, HDLc and LDLc levels, as well as, diminished NK cell and LAK cell activities. Conversely, liver SOD and NOS activities and serum TG and MDA levels were increased following CMS exposures. Administration of Banxia-houpu decoction and fluoxetine produced beneficial effects on the stressed rats by improving sucrose consumption. This behavioral change was accompanied by amelioration of numbers CMS-induced biochemical changes. Banxia-houpu decoction is a traditional Chinese prescription containing pinellia tuber, magnolia bark, hoelen, perilla herb and ginger rhizome, and has been used for centuries in China to treat mental diseases including depression and schizophrenia. However, the pharmacological profile of the decoction is different from that of fluoxetine. These findings suggest that the therapeutic actions of Banxia-houpu decoction are due to a combination of multiple biochemical effects, and may help to elucidate the mechanisms through which distinct biochemical parameters play a role in the etiology of depression.

Background Cigarette smoke a recognized risk factor for many systemic diseases and also oral diseases. Human beta defensins (HBDs), a group of important antimicrobial peptides expressed by the epithelium, are crucial for local defense and tissue homeostasis of oral cavity. The aim of this study was to evaluate potential effects of whole cigarette smoke (WCS) exposure on the expression and secretion of HBDs by oral mucosal epithelial cells. Methods Immortalized human oral mucosal epithelial (Leuk-1) cells were exposed to WCS for various time periods. HBD-1, -2 and -3 expression and subcellular localization were detected by real time qPCR, immunofluorescence assay and confocal microscopy. According to the relative fluorescent intensity, the expression levels of HBD-1, -2 and -3 were evaluated by digital image analysis system. The alteration of HBD-1, -2 and -3 secretion levels was measured by the Enzyme-Linked Immunosorbent Assay. Results WCS exposure remarkably attenuated HBD-1 expression and secretion while clearly enhanced HBD-2, -3 expression levels and HBD-2 secretion by Leuk-l cells. It appeared that there was no significant effect of WCS exposure on HBD-3 secretion. Conclusions WCS exposure could modulate expression and secretion of HBDs by oral mucosal epithelial cells, establishing a link between cigarette smoke and abnormal levels of antimicrobial peptides. The present results may give a new perspective to investigate smoking-related local defense suppression and oral disease occurrence.

Both excess dosages of drug and unwanted drug carrier can lead to severe side effects as well as the failure of tumor therapy. Here, an Fe3+–gallic acid based drug delivery system is designed for efficient monitoring of drug release in tumor. Fe3+ and polyphenol gallic acid can form polygonal nanoscale coordination polymer in aqueous solution, which exhibits certain antitumor effect. Importantly, this coordination polymer possesses extremely high doxorubicin (DOX) loading efficacy (up to 48.3%). In vitro studies demonstrate that the fluorescence of DOX can be quenched efficiently when DOX is loaded on the coordination polymer. The acidity in lysosome also triggers the release of DOX and fluorescence recovery simultaneously, which realizes real-time monitoring of drug release in tumor cells. In vivo studies further indicate that this polyphenol-rich drug delivery system can significantly inhibit tumor growth with negligible heart toxicity of DOX. This system with minimal side effects should be a promising nanoplatform for tumor treatment.

Cancer is a major public health issue, with metastatic cancer accounting for the overwhelming majority of cancer deaths. Early diagnosis and appropriate treatment of metastatic cancer may largely prolong the survival rate and improve the quality of life for patients. In this study, we have identified a panel of DNA aptamers specifically binding to MDA-MB-231 cells derived from metastatic site-pleural effusion, with high affinity after 15 rounds of selections using the cell-based systematic evolution of ligands by exponential enrichment (SELEX) method. The selected aptamers were subjected to flow cytometry and laser confocal fluorescence microscopy to evaluate their binding affinity and selectivity. The aptamer LXL-1 with the highest abundance in the enriched library demonstrated a low K(d) value and excellent selectivity for the recognition of the metastatic breast cancer cells. Tissue imaging results showed that truncated aptamer sequence LXL-1-A was highly specific to the corresponding tumor tissue and displayed 76% detection rate against breast cancer tissue with metastasis in regional lymph nodes. Therefore, on the basis of its excellent targeting properties and functional versatility, LXL-1-A holds great potential to be used as a molecular imaging probe for the detection of breast cancer metastasis. Our result clearly demonstrates that metastatic-cell-based SELEX can be used to generate DNA ligands specifically recognizing metastatic cancer cells, which is of great significance for metastatic cancer diagnosis and treatment.

An acidic polysaccharide (PY3) was isolated from the hot water extract of the red algae Porphyra yezoensis by successive column chromatographies over DEAE-cellulose and Sephadex G-200. PY3 with an average molecular weight of 1.8x10(5) was demonstrated to be composed of galactose (Gal), 3,6-anhydrogalactose (3,6-AnGal), 6-OSO3-galactose (6-OSO3-Gal) and xylose (Xyl) in an approximate molar ratio of 25 : 15, 10, 1. In view of Smith degradation and methylation and on the basis of spectral evidence including those of IR, GC, GC-MS, and H-1 and C-13 NMR, the most probable repeating unit of PY3 could be proposed as [(1-->3)beta -D-Gal(1 --> 4)alpha -L-3,6-AnGal](3)-[(1 --> 3)beta -D-Gal(1 --> 4)alpha -L-6-OSO3-Gal](2) with a xylose moiety at the C-6 of one of every twenty-five beta -D-Gal residues. To our knowledge, PY3 was shown to be the first porphyran possessing occasional xylose branches.

Although increasing studies have indicated that Nucleotide-oligomerization domain-containing protein 1 (NOD1) signaling could play an important role in gastrointestinal tumorigenesis, the protein expression and function of NOD1 signaling have not been understood well in oral squamous cell carcinoma (OSCC) progression. The objective of this study is, thus, to examine protein expression of NOD1 signaling immunohistochemically in normal, premalignant and malignant specimens of oral cavity, and to take insights into the association between the protein expression of NOD1 signaling pathway and OSCC precession. In this study immunohistochemical expression of NOD1, Receptor-interacting protein 2 (RIP2), Caspase12, human β Defensin1, 2 and 3 (hBD1, 2, 3) was examined in 15 normal controls, 30 cases of oral leukoplakia (OLK) and 60 cases of OSCC. The immunostaining score was assessed by 2 pathologists, respectively. We found that the expression of NOD1, RIP2, Caspase12, hBD1, 2, and 3 decreased along with the progression of OSCC. NOD1 expression was correlated significantly to tumor differentiation, lymph node metastasis, and tumor size. Our results also showed the correlation of hBD2, 3 to lymph node metastasis of OSCC. These results suggest that the dysfunction of NOD1 signaling pathways could be associated with OSCC development and progression. NOD1, RIP2 and Caspase12 could be used as potentially novel biomarkers for oral carcinogenesis.

Sesaminol is one component of sesame oil and has been widely used as the stabilizer to extend the storage period of food oil in China. In this study, we tried to investigate the antioxidant activity of sesaminol on rat pheochromocytoma (PC12) cells oxidative damaged by H2O2. Cell viability, LDH level and apoptosis of the PC12 cells were assayed after treatment with sesaminol for 3 h and exposure to H2O2. Furthermore, superoxide (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and intracellular ROS were assayed after exposure of the PC12 cells to H2O2. The results showed that pre-treatment with sesaminol prior to H2O2 exposure significantly elevated cell survival rate and SOD, CAT and GSH-Px activity. Meanwhile, sesaminol declined the secreted LDH level, apoptosis rate and ROS level of H2O2 exposed cells. Thus, sesaminol may protect PC12 against oxidative injury. Copyright © 2012 John Wiley & Sons, Ltd.

Cigarette smoke exposure is associated with increased risk of different disorders. Immunological dysfunction especially in macrophages is one of important reasons in the initiation, progression and exacerbation of smoke-related pulmonary illnesses. However, it is still obscure how cigarette smoke impacts the vitality and functions of macrophages. In the present study, we examined the effects of cigarette smoke extract (CSE) on mouse Ana-1 macrophages and tried to elucidate the involved mechanism. The results showed CSE induced cell apoptosis accompanied by increased releasing of lactate dehydrogenase (LDH), mitochondrial injury and oxidative stress. It also inhibited anti-apoptosis protein Bcl-2 expression and promoted pro-apoptosis protein Bax and Bad expressions. Moreover, low-dose CSE increased nuclear NF-κB levels of macrophages; on the contrary, high-dose CSE or long-time treatment decreased it. These observations were in correspondence with changes of intracellular ROS level and antioxidant enzymes’ activity. Furthermore, pretreatment with 10 μM of NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) for 1 h significantly enhanced macrophage apoptosis. Taken together, these data implied that mitochondrial dysfunction and oxidative stress played important roles in the injury of Ana-1 cells caused by CSE, which was related to NF-κB pathway; an anti-apoptotic program played a dominant role at low doses/short-term exposure to CSE, whereas a pro-apoptotic program was initiated at high doses/long-term exposure.

Sesaminol is one component of sesame oil and has been widely used as the stabilizer to extend the storage period of food oil in China. In this study, we tried to investigate the antioxidant activity of sesaminol on rat pheochromocytoma (PC12) cells oxidative damaged by H2 O2 . Cell viability, LDH level and apoptosis of the PC12 cells were assayed after treatment with sesaminol for 3 h and exposure to H2 O2 . Furthermore, superoxide (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and intracellular ROS were assayed after exposure of the PC12 cells to H2 O2 . The results showed that pre-treatment with sesaminol prior to H2 O2 exposure significantly elevated cell survival rate and SOD, CAT and GSH-Px activity. Meanwhile, sesaminol declined the secreted LDH level, apoptosis rate and ROS level of H2 O2 exposed cells. Thus, sesaminol may protect PC12 against oxidative injury.

BackgroundAnterior gradient homolog 2 (AGR2) is a functional protein with critical roles in a diverse range of biological systems, including vertebrate tissue development, inflammatory tissue injury responses, and cancer progression. Clinical studies have shown that the AGR2 protein is overexpressed in a wide range of human cancers, including carcinomas of the esophagus, pancreas, breast, prostate, and lung, making the protein as a potential cancer biomarker. However, the general biochemical functions of AGR2 in human cells remain undefined, and the signaling mechanisms that drive AGR2 to inhibit p53 are still not clearly illustrated. Therefore, it is of great interest to develop molecular probes specifically recognizing AGR2 for its detection and for the elucidation of AGR2-associated molecular mechanism.Methodology/principal findingsThrough a bead-based and flow cytometry monitored SELEX technology, we have identified a group of DNA aptamers that can specifically bind to AGR2 with K(d) values in the nanomolar range after 14 rounds of selections. Aptamer C14B was chosen to further study, due to its high binding affinity and specificity. The optimized and shortened C14B1 has special G-rich characteristics, and the G-rich region of this binding motif was further characterized to reveal an intramolecular parallel G-quadruplex by CD spectroscopy and UV spectroscopy. Our experiments confirmed that the stability of the G-quadruplex structure was strongly dependent on the nature of the monovalent ions and the formation of G-quadruplex structure was also important for the binding capacity of C14B1 to the target. Furthermore, we have designed a kind of allosteric molecule beacon (aMB) probe for selective and sensitive detection of AGR2.Conclusion/significanceIn this work, we have developed new aptamer probes for specific recognition of the AGR2. Structural study have identified that the binding motif of aptamer is an intramolecular parallel G-quadruplex structure and its structure and binding affinity are strongly dependent on the nature of the monovalent ion. Furthermore, with our design of AGR2-aMB, AGR2 could be sensitively and selectively detected. This aptamer probe has great potential to serve as a useful tool for early diagnosis and prognosis of cancer and for fundamental research to elucidate the biochemical functions of AGR2.

BACKGROUND: Glioblastoma is the most common and most lethal form of brain tumor in human. Unfortunately, there is still no effective therapy to this fatal disease and the median survival is generally less than one year from the time of diagnosis. Discovery of ligands that can bind specifically to this type of tumor cells will be of great significance to develop early molecular imaging, targeted delivery and guided surgery methods to battle this type of brain tumor. METHODOLOGY/PRINCIPAL FINDINGS: We discovered two target-specific aptamers named GBM128 and GBM131 against cultured human glioblastoma cell line U118-MG after 30 rounds selection by a method called cell-based Systematic Evolution of Ligands by EXponential enrichment (cell-SELEX). These two aptamers have high affinity and specificity against target glioblastoma cells. They neither recognize normal astraglial cells, nor do they recognize other normal and cancer cell lines tested. Clinical tissues were also tested and the results showed that these two aptamers can bind to different clinical glioma tissues but not normal brain tissues. More importantly, binding affinity and selectivity of these two aptamers were retained in complicated biological environment. CONCLUSION/SIGNIFICANCE: The selected aptamers could be used to identify specific glioblastoma biomarkers. Methods of molecular imaging, targeted drug delivery, ligand guided surgery can be further developed based on these ligands for early detection, targeted therapy, and guided surgery of glioblastoma leading to effective treatment of glioblastoma.

Dendritic cells (DCs) play an important role in initiating antitumour immune response. Tumour progression usually induces defects in DC maturation and thus tumour-bearing hosts exhibit immunosuppression and tumour escape. The previous studies showed that an exopolysaccharide (EPS) from a fungus, one anamorph of Cordyceps sinensis, inhibited tumour growth via activating immune response in the hosts. In view of the crucial actions of DCs in antitumour immunity, the present study aims to explore the effects of EPS on murine DCs. Murine DCs were derived from the bone marrow of C57BL/6 mice, and the effects of EPS on phenotype molecules and ingestion function of DCs were assayed using flow cytometry. Cytokine expressions of DCs were assayed by reverse transcriptase-PCR. Additionally, the level of phosphorylated signal transducers and activators of transcription 3 (p-STAT3) of DCs was evaluated using Western blotting. The results showed that EPS promoted the levels of surface molecules MHC II, CD40, CD80 and CD86 of DCs and decreased their ingestion ability. The mRNA expressions of cytokines (IL-12p40 and TNF-α) and inducible nitric oxide synthase were up-regulated by EPS. We also found that EPS significantly down-regulated p-STAT3 level of DCs. The results suggested that the promotion of DC's maturation and activation by EPS is probably related to the inhibition of STAT3 phosphorylation.