A fluorescence sensing platform of theophylline based on the interaction of RNA aptamer with graphene oxide.

RNA, with a structure similar to DNA, should exhibit similar behaviors when it interacts with graphene. In this work, we designed a sensing platform of theophylline based on the interaction of an RNA aptamer with graphene oxide (GO) using the fluorescence as a sensing signal. Firstly, quantum dots (QDs) were modified with the selected ssRNA that can be used as an aptamer to recognize the theophylline. The fluorescence of QDs will be quenched in the presence of GO due to the noncovalent assembly between ssRNA aptamer and GO, leading to fluorescence resonance energy transfer (FRET) from QDs to GO, fluorescence "turn-off". Then, in the presence of theophylline, the ssRNA aptamer recognizes theophylline to form a dsRNA-theophylline complex. The weak affinity between the complex and GO makes QDs move away from the GO surface, leading to the fluorescence recovery of QDs, fluorescence "turn-on". Because of the high fluorescence quenching efficiency, unique structure of GO and specificity of the RNA aptamer, the proposed sensing platform exhibits high sensitivity and excellent selectivity for the determination of theophylline. The excellent performance of the sensor based on GO provides new opportunities for sensitive and selective detection of biorecognition events.

Synergistic antitumoral activity and induction of apoptosis by novel pan Bcl-2 proteins inhibitor apogossypolone with adriamycin in human hepatocellular carcinoma.

AIM: To investigate the in vitro and in vivo activities and related mechanism of apogossypolone (ApoG2) alone or in combination with adriamycin (ADM) against human hepatocellular carcinoma (HCC). METHODS: The IC50 of ApoG2 in vitro was tested by WST assay, and the synergistic effect was analyzed using the CalcuSyn method. Cell apoptosis was determined using 4',6-diamidino-2- phenylindole staining and flow cytometric analysis. Western blotting was used to determine the expression of apoptosis-related proteins. In vivo activity was evaluated in the xenograft model in nude mice, and apoptosis in tumor tissues was determined by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling (TUNEL) assay. RESULTS: The IC50 of ApoG2 in HCC cells was 17.28-30.63 micromol/L. When ApoG2 was combined with ADM, increased cytotoxicity and apoptosis were observed in SMMC-7721 cells compared to treatment with ApoG2 alone. The Western blotting results indicated that the ApoG2 induced apoptosis in SMMC-7721 cells by downregulating anti-apoptotic proteins Bcl-2, Mcl-1, and Bcl-XL, up-regulating pro-apoptotic protein Noxa, and promoting the activities of caspases-9 and -3. The tumor growth of xenograft SMMC-7721 was inhibited in nude mice when ApoG2 was administered orally without causing damage to the normal tissues. The in vivo study also indicated an increasing anti-tumoral effect when ApoG2 at 100 or 200 mg/kg dosages were used together with ADM at 5.5 mg/kg, with relative tumor proliferation rate (T/C) values of 0.456 and 0.323, respectively. Apoptosis induced in vivo by ApoG2 alone or combined with ADM was confirmed by TUNEL assay in tumor tissues. CONCLUSION: ApoG2 is a potential non-toxic target agent that induces apoptosis by upregulating Noxa, while inhibiting anti-apoptotic proteins and promoting the effect of chemotherapy agent ADM in HCC.

XIAP Limits Autophagic Degradation of Sox2 and Is A Therapeutic Target in Nasopharyngeal Carcinoma Stem Cells.

Rationale: Nasopharyngeal carcinoma (NPC) is the most frequent head and neck tumor in South China. The presence of cancer stem cells (CSCs) in NPC contributes to tumor maintenance and therapeutic resistance, while the ability of CSCs to escape from the apoptosis pathway may render them the resistant property to the therapies. Inhibitor of apoptosis proteins family proteins (IAPs), which are overexpressed in nasopharyngeal carcinoma stem cells, may play an important role in maintaining nasopharyngeal cancer stem cell properties. Here, we develop a novel CSC-targeting strategy to treat NPC through inhibiting IAPs. Methods: Human NPC S-18 and S-26 cell lines were used as the model system in vitro and in vivo. Fluorescence activated cell sorting (FACS) assay was used to detect nasopharyngeal SP cells and CD44+ cells. The characteristics of CSCs were defined by sphere suspension culture, colony formation assay and cell migration. The role of XIAP on the regulation of Sox2 protein stability and ERK1-mediated phosphorylation of Sox2 signaling pathway were analyzed using immunoblotting, immunoprecipitation, immunofluorescence, phosphorylation mass spectrometry, siRNA silencing and plasmid overexpression. The correlation between XIAP and Sox2 in NPC biopsies and their role in prognosis was performed by immunohistochemistry. APG-1387 or chemotherapies-induced cell death and apoptosis in S-18 and S-26 were determined by WST, immunoblotting and flow cytometry assay. Results: IAPs, especially X chromosome-linked IAP (XIAP), were elevated in CSCs of NPC, and these proteins were critically involved in the maintenance of CSCs properties by enhancing the stability of Sox2. Mechanistically, ERK1 kinase promoted autophagic degradation of Sox2 via phosphorylation of Sox2 at Ser251 and further SUMOylation of Sox2 at Lys245 in non-CSCs. However, XIAP blocked autophagic degradation of Sox2 by inhibiting ERK1 activation in CSCs. Additionally, XIAP was positively correlated with Sox2 expression in NPC tissues, which were associated with NPC progression. Finally, we discovered that a novel antagonist of IAPs, APG-1387, exerted antitumor effect on CSCs. Also, the combination of APG-1387 with CDDP /5-FU has a synergistic effect on NPC. Conclusion: Our study highlights the importance of IAPs in the maintenance of CSCs in NPC. Thus, XIAP is a promising therapeutic target in CSCs and suggests that NPC patients may benefit from a combination treatment of APG-1387 with conventional chemotherapy.

Identification of novel inhibitors of the streptogramin group A acetyltransferase via virtual screening.

Virginiamycin acetyltransferase D (VatD) plays a vital rule in streptogramins resistance by chemically inactivating streptogramin A. Therefore, it is desirable to discover novel small molecular weight inhibitors of VatD via state-of-the-art virtual screening techniques. This "cocktail" strategy by combining VatD inhibitor with streptogramins may provide new therapeutic opportunity for resistant bacteria infections. Structure-based virtual screening method (molecular docking) was applied to rank and score a chemical database containing 300 000 commercially available compounds against the VatD substrate binding site. Twenty six out of the 200 top scored compounds from the docking calculation were selected and submitted to the VatD enzymatic inhibition assay. The plasmid pRSET B/vatD was constructed and transformed into E. coli (trxB) host cells for over-expression, and VatD enzyme was purified and validated by showing acetyltransferase activity to Virginiamycin M1. Three out of these 26 tested compounds showed enzymatic inhibition on VatD with IC50 168.6, 91.0 and 55.2 micromol x L(-1), separately. Other compounds could not be dissolved in the system and/or had little effect on the enzyme (IC50 > 200 micromol x L(-1)). To our knowledge, it is first time that small molecular weight organic compounds were identified as VatD inhibitors. It is expected that the VatD inhibitors identified at present study could serve as lead compounds for the further development of the novel therapeutic agents to overcome streptogramins resistance.

Electrocatalysis of oxygen at hemoglobin-Au colloid-1,4-benzenedimethanethiol modified electrode.

A novel renewable O2 sensor based on the direct electron transfer of hemoglobin (Hb) is proposed. Hb was immobilized on a gold nanoparticles (GNP) associated with a 1,4-benzenedimethanethiol (BDT) monolayer which were modified the electrode. The direct electrochemistry of Hb was investigated by electrochemical methods and cyclic voltammetric showing a pair of redox peaks of Hb. The high efficiency of the Hb/GNP/BDT modified gold electrode towards the catalytic electro-reduction of oxygen has been observed and the potential application of Hb/GNP/BDT modified gold electrode as biosensors to monitor O2 is proposed. The electrocatalytic response showed a linear dependence on the O2 concentration ranging from 2.0 to 40.0 micromol/L.

Electrochemical determination of 6-mercaptopurine at silver microdisk electrodes.

The electrochemical behaviour of 6-mercaptopurine (6-MP) at a microdisk electrode is investigated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The results indicate that 6-MP can be strongly adsorbed on the surface of the static mercury drop electrode (SMDE) and reacts with Ag+ ions which are produced at positive potentials. 6-MP yields a well-defined cathodic stripping signal during the negative scan at about -0.812 V (vs. SCE) in pH 9.0 phosphate buffer solution. The electrode has hence been used for the determination of 6-MP by differential pulse voltammetry (DPV). The linear range is between 2.0x10(-7) and 5.0x10(-5) mol/l, with the calculated detection limit (S/N=3) of 8.0x10(-8) mol/l. The relative standard deviation is 3.0% for eight successive determinations of 4.0x10(-5) mol/l 6-MP. The determination of 6-MP in tablets has also been carried out and satisfactory results have been obtained.

Electrocatalytical oxidation of hydroquinone with ferrocene covalently bound to L-cysteine self-assembled monolayers on a gold electrode.

The preparation of a gold electrode modified by ferrocenecarboxylic acid (FcA) covalently bound to L-cysteine self-assembled monolayer (FcA-SAM) is described. The modified electrode shows an excellent electrocatalytic activity for the oxidation of hydroquinone (QH2) and accelerates the electron transfer rate. The anodic overpotential is reduced by ca. 290 mV compared to those obtained at bare gold electrodes. The charge transfer coefficient and the apparent surface electron transfer rate constant for the redox couple of Q/QH2 at the modified electrode are found to be 0.425 and 0.96 s(-1), respectively. The catalytic current response of DPV increases linearly with the QH2 concentration from 5.7 x 10(-7) to 3.2 x 10(-4) M. The estimation of QH2 in a simulative sample is satisfactory. The method is simple, quick, and sensitive.

Smac mimetics in combination with TRAIL selectively target cancer stem cells in nasopharyngeal carcinoma.

Nasopharyngeal carcinoma is a common malignancy in Southern China. After radiotherapy and chemotherapy, a considerable proportion of patients with nasopharyngeal carcinoma suffered tumor relapse and metastasis. Cancer stem cells (CSC) have been shown with resistance against therapies and thus considered as the initiator of recurrence and metastasis in tumors, where the antiapoptotic property of CSCs play an important role. Smac/DIABLO is an inverse regulator for the inhibitors of apoptosis protein family (IAP), which have been involved in apoptosis. Here, the effects of Smac mimetics on the CSCs of nasopharyngeal carcinoma were studied both in vitro and in vivo, using two clones of nasopharyngeal carcinoma cell line CNE2 as models. We found that one of the clones, S18, had CSC-like properties and IAPs were overexpressed. The combination of Smac mimetics and TNF-related apoptosis-inducing ligand (TRAIL) can reduce the percentage of SP cells and inhibit the colony- and sphere-forming abilities of S18 cells, indicating their ability to attenuate the CSCs. Moreover, in a nasopharyngeal carcinoma xenograft model, the administration of Smac mimetics in combination with TRAIL also led to the elimination of nasopharyngeal carcinoma stem cells. Furthermore, the Smac mimetics in combination with TRAIL induced the degradation of cIAP1 and XIAP and thus induced apoptosis in vitro and in vivo. Taken together, our data show that Smac mimetics exerted an antitumor effect on nasopharyngeal carcinoma cancer stem cells, and this combination treatment should be considered as a promising strategy for the treatment of nasopharyngeal carcinoma.

Modification of conductive polymer for polymeric anodes of flexible organic light-emitting diodes.

A conductive polymer, poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS), was modified with dimethyl sulfoxide (DMSO) in solution state, together with sub-sequential thermal treatment of its spin-coated film. The electrical conductivity increased by more than three orders of magnitude improvement was achieved. The mechanism for the conductivity improvement was studied at nanoscale by particle size analysis, field emission scanning electron microscopy (FESEM), and X-ray photoelectron spectroscopy (XPS). Smaller particle size was observed, resulting in larger contact area and better electrical conductive connections. Connection of conductive PEDOT increased on the surface of the PEDOT:PSS particles, which promoted high conductivity. Flexible anodes based on the modified PEDOT:PSS were fabricated. Flexible organic light-emitting diodes (FOLED) based the polymeric anodes have a comparable performance to those on indium-tin-oxide (ITO) anodes.

Flexible organic light-emitting diodes with a polymeric nanocomposite anode.

Flexible organic light-emitting diodes (FOLEDs) are facing mechanical issues arising from failure of the indium-tin oxide (ITO) films fabricated on flexible substrates. Polymeric nanocomposite anodes were fabricated by including single-wall carbon nanotubes (SWCNTs) in aqueous poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS). The conductivity, transmittance and flexibility of the polymeric nanocomposite anode were characterized. The polymeric nanocomposite anodes fabricated on a poly(ethylene terephthalate) (PET) substrate exhibited superior bending properties to ITO anodes on PET. The FOLEDs fabricated on the polymeric nanocomposite anodes had a low turn-on voltage and higher luminous intensity than those fabricated on ITO/PET anodes. This flexible nanocomposite polymeric anode is a very promising for fully FOLEDs and other optoelectronics.

Preparation of novel mercury-doped silver nanoparticles film glassy carbon electrode and its application for electrochemical biosensor.

A novel mercury-doped silver nanoparticles film glassy carbon (Ag/MFGC) electrode was prepared in this study. Electrochemical behaviors of cysteine on the Ag/MFGC electrode were investigated by electrochemical impedance spectroscopy and cyclic voltammetry (CV). The results indicated that cysteine could be strongly adsorbed on the surface of the Ag/MFGC electrode to form a thin layer. The doped electrode could catalyze the electrode reaction process of cysteine, and the cysteine displayed a pair of well-defined and nearly reversible CV peaks at the electrode in an acetate buffer solution (pH 5.0). The Ag/MFGC electrode was used for determination of cysteine by differential pulse voltammetry. The linear range was between 4.0x10(-7) and 1.3x10(-5) mol/L, with a detection limit of 1.0x10(-7) mol/L and a signal-to-noise ratio of 3. The relative standard deviation was 2.4% for seven successive determinations of 1.0x10(-5) mol/L cysteine. The determinations of cysteine in synthetic samples and urinal samples were carried out and satisfactory results were obtained. Amperometric application of the Ag/MFGC electrode as biosensors is proposed.