Solvated Electrons Generated on the Surface of Na-SPHI for Boosting Visible Light Photocatalytic Hydrogen Evolution with Ultra-High AQE.

Enhancing the utilization of visible-light-active semiconductors with an excellent apparent quantum efficiency (AQE) remains a significant and challenging goal in the realm of photocatalytic water splitting. In this study, a fully condensed sulfur-doped poly(heptazine imide) metalized with Na (Na-SPHI) is synthesized by an ionothermal method by using eutectic NaCl/LiCl mixture as the ionic solvent. Comprehensive characterizations of the obtained Na-SPHI reveal several advantageous features, including heightened light absorption, facilitated exciton dissociation, and expedited charge transfer. More importantly, solvated electron, powerful reducing agents, can be generated on the surface of Na-SPHI upon irradiation with visible light. Benefiting from above advantage, the Na-SPHI exhibits an excellent H2 evolution rate of 571.8 µmol·h-1 under visible light illumination and a super-high AQE of 61.7% at 420 nm. This research emphasizes the significance of the solvated electron on the surface of photocatalyst in overcoming the challenges associated with visible light-driven photocatalysis, showcasing its potential application in photocatalytic water splitting.

Manganese-Implanted Titanium Modulates the Crosstalk between Bone Marrow Mesenchymal Stem Cells and Macrophages to Improve Osteogenesis.

Manganese (Mn) is an essential micronutrient in various physiological processes, but its functions in bone metabolism remain undefined. This is partly due to the interplay between immune and bone cells because Mn plays a central role in the immune system. In this study, we utilized the plasma immersion ion implantation and deposition (PIII&D) technique to introduce Mn onto the titanium surface. The results demonstrated that Mn-implanted surfaces stimulated the shift of macrophages toward the M1 phenotype and had minimal effects on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells (mBMSCs) under mono-culture conditions. However, they promoted the M2 polarization of macrophages and improved the osteogenic activities of mBMSCs under co-culture conditions, indicating the importance of the crosstalk between mBMSCs and macrophages mediated by Mn in osteogenic activities. This study provides a positive incentive for the application of Mn in the field of osteoimmunology.

Establishment of protoplasts transient expression system in Pinellia ternata (Thunb.) Breit.

OBJECTIVE: In this study, we established an efficient and rapid transient expression system in the protoplasts of Pinellia ternata (Thunb.) Breit. (P. ternata). RESULTS: The protoplasts of P. ternata were prepared from plant leaves as the source material by digesting them with the combination of 20 g·l-1 cellulase and 15 g·l-1 macerozyme for 6 h. Based on the screening of PEG concentration, the conditions for PEG-mediated protoplast transformation were improved, and the highest transformation efficiency was found for 40% PEG 4000. Furthermore, we used the subcellular protein localization technique in P. ternata protoplasts to allow further validation of transient expression system. CONCLUSIONS: We present the method that can be applicable for studying both gene verification and expression in P. ternata protoplasts, thus allowing for engineering the improved varieties of P. ternata through molecular plant breeding techniques. This method can also be widely applicable for analyzing protein interactions, detecting promoter activity, for somatic cell fusion in plant breeding, as well as for other related studies.

Copper-Zinc Bimetallic Single-Atom Catalysts with Localized Surface Plasmon Resonance-Enhanced Photothermal Effect and Catalytic Activity for Melanoma Treatment and Wound-Healing.

Nanomaterials with photothermal combined chemodynamic therapy (PTT-CDT) have attracted the attention of researchers owing to their excellent synergistic therapeutic effects on tumors. Thus, the preparation of multifunctional materials with higher photothermal conversion efficiency and catalytic activity can achieve better synergistic therapeutic effects for melanoma. In this study, a Cu-Zn bimetallic single-atom (Cu/PMCS) is constructed with augmented photothermal effect and catalytic activity due to the localized surface plasmon resonance (LSPR) effect. Density functional theory calculations confirmed that the enhanced photothermal effect of Cu/PMCS is due to the appearance of a new d-orbital transition with strong spin-orbit coupling and the induced LSPR. Additionally, Cu/PMCS exhibited increased catalytic activity in the Fenton-like reaction and glutathione depletion capacity, further enhanced by increased temperature and LSPR. Consequently, Cu/PMCS induced better synergistic anti-melanoma effects via PTT-CDT than PMCS in vitro and in vivo. Furthermore, compared with PMCS, Cu/PMCS killed bacteria more quickly and effectively, thus facilitating wound healing owing to the enhanced photothermal effect and slow release of Cu2+ . Cu/PMCS promoted cell migration and angiogenesis and upregulated the expression of related genes to accelerate wound healing. Cu/PMCS has potential applications in treating melanoma and repairing wounds with its antitumor, antibacterial, and wound-healing properties.

Optimizing the band structure of sponge-like S-doped poly(heptazine imide) with quantum confinement effect towards boosting visible-light photocatalytic H2 generation.

Simultaneously manipulating the nanostructure and band structure of semiconductors for boosting the photocatalytic performance of photocatalyts is highly desirable. Herein, a series of hierarchical sponge-like S-doped poly(heptazine imide) (HS-SPHI) assembled by ultrathin nanosheets were successfully fabricated via a facile bottom-up supramolecular preassembly approach using melamine (MA) and trithiocyanuric acid (TTCA) as precursors. Benefiting from the synergistic effect of the S-doping and their unique hierarchical porous structure coupled with quantum confinement effect, the as-obtained HS-SPHIs are endowed with extended visible-light response, improved charge separation efficiency, enlarged specific surface area, and enhanced thermodynamic driving force for water reduction. As a result, all the HS-SPHIs exhibit remarkable boosting visible-light (>420 nm) photocatalytic H2evolution (PHE). The maximum PHE rate achieved by HS-SPHI-650 can be up to 3584.2 µmol g-1h-1, with an apparent quantum efficiency (AQE) of 14.67 % at 420 nm, which is about 22.4 times than that of pristine bulk g-C3N4 (B-GCN). We believe that this work will provide a significant strategy for optimizing the band structure of PCN in order to improve its photocatalytic performance.

Transcriptome revealing the dual regulatory mechanism of ethylene on the rhynchophylline and isorhynchophylline in Uncaria rhynchophylla.

Rhynchophylline (RIN) and isorhynchophylline (IRN) are extracted from Uncaria rhynchophylla, which are used to treat Alzheimer's disease. However, the massive accumulation of RIN and IRN in U. rhynchophylla requires exogenous stimulation. Ethylene is a potential stimulant for RIN and IRN biosynthesis, but there is no study on the role of ethylene in RIN or IRN synthesis. This study investigated the regulation of ethylene in RIN and IRN biosynthesis in U. rhynchophylla. An increase in the content of RIN and IRN was observed that could be attributed to the release of ethylene from 18 mM ethephon, while ethylene released from 36 mM ethephon reduced the content of RIN and IRN. The transcriptome and weighted gene co-expression network analysis indicated the up-regulation of seven key enzyme genes related to the RIN/IRN biosynthesis pathway and starch/sucrose metabolism pathway favored RIN/IRN synthesis. In comparison, the down-regulation of these seven key enzyme genes contributed to the reduction of RIN/IRN. Moreover, the inhibition of photosynthesis is associated with a reduction in RIN/IRN. Photosynthesis was restrained owing to the down-regulation of Lhcb1 and Lhcb6 after 36 mM ethephon treatment and further prevented supply of primary metabolites (such as α-D-glucose) for RIN/IRN synthesis. However, uninterrupted photosynthesis ensured a normal supply of primary metabolites at 18 mM ethephon treatment. AP2/ERF1, bHLH1, and bHLH2 may positively regulate the RIN/IRN accumulation, while NAC1 may play a negative regulatory role. Our results construct the potential bidirectional model for ethylene regulation on RIN/IRN synthesis and provide novel insight into the ethylene-mediated regulation of the metabolism of terpenoid indole alkaloids.

Band structure engineering of PTI in C-PTI/ZnO heterostructures for enhanced visible-light-driven H2 evolution.

Polytriazine imide (PTI), a triazine-based carbon nitride has a wider band gap and more positive conduction band (CB) potential compared to those of graphitic carbon nitride (g-C3N4). Therefore, it is highly desired to develop an effective strategy to optimize the band structure of PTI for the enhancement of the photocatalytic performance, especially upshift the conductive band potential. Here, a ternary C-PTI/ZnO (CPZ) photocatalyst was developed via a simple one-step molten salt method. In the obtained CPZ sample, the carbon ring in-plane connects to the triazine ring, leading to the formation of C-PTI nanosheets. The carbon ring incorporation not only efficiently narrows the band gap of PTI, but also shifts its conduction band potential negatively and accelerates the photogenerated electron transport. In addition, ZnO nanoparticles are well dispersed on the C-PTI nanosheets, further promoting the charge carriers transfer and separation. As a result, the CPZ sample presents a photocatalytic H2 evolution rate up to 52 µmol h-1 under visible light, which is 60 and 179 times higher than that of C-PTI and PTI, respectively.

Highly sensitive fluorescent bioassay of 2,3,7,8-tetrachloro-dibenzo-p-dioxin based on abnormal expression of cytochrome P450 1A2 in human cells.

Current in vitro bioassays of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, a major threat carcinogen) are relied on murine cells and fluorescent probe 7-ethoxyresorufin (7-ER), in which TCDD mostly causes abnormal expression of cytochrome P450 1A1 (CYP1A1). However, for human cells, TCDD mainly leads to a distinct abnormal expression of cytochrome P450 1A2 (CYP1A2). The poor response of 7-ER to CYP1A2 limits the traditional bioassay for human cells. Herein, we report a fluorescent probe N-(3-hydroxybutyl)-4-methoxy-1,8-naphthalimide (HBMN) for in vitro bioassay of TCDD with human cells. HBMN had ca. 60 times higher affinity to CYP1A2 than 7-ER. As such, the sensing sensitivity increased by 10 times, and different expression of CYP1A2 by TCDD induction in different human cells was found. Besides, HBMN was also feasible in rapid screening of TCDD concentration by naked eye. It would open a new way to highly sensitive detect TCDD and understand the pathogenesis of TCDD in different human organs.

Ultrasensitive electrochemical detection of poly (ADP-ribose) polymerase-1 via polyaniline deposition.

Recent findings have thrust poly ADP (ADP: adenosine diphosphate)-ribose polymerase-1 (PARP-1) into the limelight as potential chemotherapeutic target because it is closely related to the development of tumor. So, studies on its detection and inhibitors evaluation have attracted more attention. It is interesting that poly (ADP-ribose) (PAR), the catalytic product of PARP-1 in the existence of nicotinamide adenine dinucleotide (NAD+), possess twice charge density of DNA strands. PAR contain 200 units, i.e., about 400bp bases, and multiple branched strands. So, plentiful negative charges on PAR supplied exquisite environment for PANI deposition, which was triggered by horseradish peroxidase (HRP). Because of the unique electrochemical property of PANI, ultrasensitive electrochemical detection of PARP-1 was proposed. Under optimum conditions, DPV intensity linearly increased with the increment of PARP-1 in the range of 0.005-1.0 U. The detection limit was 0.002 U, which was comparable or more sensitive than that obtained from previously reported strategies.

Electrochemiluminescent detection of cardiac troponin I by using soybean peroxidase labeled-antibody as signal amplifier.

This work proposed an electrochemiluminescence (ECL) immunosensor for quantitative monitoring of cardiac troponin I (cTnI) in plasma with soybean peroxidase (SBP) labeled-antibody as signal amplifier. The ECL sandwich immunosensor was constructed by covalent binding anti-cTnI capture antibody (Ab1) to polyethylenimine-functionalized graphene matrix, which was obtained by a simple hydrothermal reaction between polyethylenimine (PEI) and graphene oxide (GO). After that, the SBP-labeled detection antibody (SBP-Ab2), synthesized by NaIO4 method, was immobilized on the surface of electrode through sandwich immunoreaction. The SBP on electrode surface displayed strong and stable ECL signal of luminol in the presence of H2O2, which could be used for cTnI detection with a concentration range of 5-30,000pg/mL and a detection limit of 3.3pg/mL. This proposed SBP-modified immunosensor displayed high sensitivity, selectivity and accuracy, and was expected not only to detect cTnI in practical human plasma sample but also to be used in other biomarkers detection.

In Situ Detection and Imaging of Telomerase Activity in Cancer Cell Lines via Disassembly of Plasmonic Core-Satellites Nanostructured Probe.

The label-free localized surface plasmon resonance (LSPR) detection technique has been identified as a powerful means for in situ investigation of biological processes and localized chemical reactions at single particle level with high spatial and temporal resolution. Herein, a core-satellites assembled nanostructure of Au50@Au13 was designed for in situ detection and intracellular imaging of telomerase activity by combining plasmonic resonance Rayleigh scattering spectroscopy with dark-field microscope (DFM). The Au50@Au13 was fabricated by using 50 nm gold nanoparticles (Au50) as core and 13 nm gold nanoparticles (Au13) as satellites, both of them were functionalized with single chain DNA and gathered proximity through the highly specific DNA hybridization with a nicked hairpin DNA (O1) containing a telomerase substrate (TS) primer as linker. In the presence of telomerase, the telomeric repeated sequence of (TTAGGG)n extended at the 3'-end of O1 would hybridized with its complementary sequences at 5'-ends. This led the telomerase extension product of O1 be folded to form a rigid hairpin structure. As a result, the Au50@Au13 was disassembled with the releasing of O1 and Au13-S from Au50-L, which dramatically decreased the plasmon coupling effect. The remarkable LSPR spectral shift was observed accompanied by a detectable color change from orange to green with the increase of telomerase activity at single particle level with a detection limit of 1.3 × 10-13 IU. The ability of Au50@Au13 for in situ imaging intracellular telomerase activity, distinguishing cancer cells from normal cells, in situ monitoring the variation of cellular telomerase activity after treated with drugs were also demonstrated.

Confining a bi-enzyme inside the nanochannels of a porous aluminum oxide membrane for accelerating the enzymatic reactions.

An artificial metabolon with high conversion efficiency was constructed by confining a bi-enzyme into porous aluminum oxide nanochannels, which accelerated enzymatic reactions by minimizing the diffusion loss of intermediate species.

Construction of iron-polymer-graphene nanocomposites with low nonspecific adsorption and strong quenching ability for competitive immunofluorescent detection of biomarkers in GM crops.

We developed a new immunofluorescent biosensor by utilizing a novel nanobody (Nb) and iron-polymer-graphene nanocomposites for sensitive detection of 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacdterium tumefaciens strain CP4 (CP4-EPSPS), which considered as biomarkers of genetically modified (GM) crops. Specifically, we prepared iron doped polyacrylic hydrazide modified reduced graphene nanocomposites (Fe@RGO/PAH) by in-situ polymerization approach and subsequent a one-pot reaction with hydrazine. The resulting Fe@RGO/PAH nanocomposites displayed low nonspecific adsorption to analytes (11% quenching caused by nonspecific adsorption) due to electrostatic, energetic and steric effect of the nanocomposites. After Nb immobilizing, the as-prepared Fe@RGO/PAH/Nbs showed good selectivity and high quenching ability (92% quenching) in the presence of antigen (Ag) and polyethylene glycol (PEG) modified CdTe QDs (Ag/QDs@PEG), which is a nearly 4 fold than that of the unmodified GO in same condition. The high quenching ability of Fe@RGO/PAH/Nbs can be used for detection of CP4-EPSPS based on competitive immunoassay with a linearly proportional concentration range of 5-100ng/mL and a detection limit of 0.34ng/mL. The good stability, reproducibility and specificity of the resulting immunofluorescent biosensor are demonstrated and might open a new window for investigation of fluorescent sensing with numerous multifunctional graphene based materials.

A portable chemiluminescence imaging immunoassay for simultaneous detection of different isoforms of prostate specific antigen in serum.

A multianalyte chemiluminescence (CL) imaging immunoassay strategy for sensitive detection of different isoforms of prostate specific antigen (PSA) was developed. The microtiter plates were fabricated by simultaneously immobilizing of free-PSA (f-PSA) and total-PSA (t-PSA) capture antibody on nitrocellulose (NC) membrane. Each of the array were spotted in replicates of six spots within a spacing of 2mm. 16 or 48 detection wells were integrated on a single NC membrane and each well could be used as a microreactor and microanalysis chamber. Under a sandwiched immunoassay, the CL signals on each sensing site were collected by a charge-coupled device (CCD), presenting an array-based chemiluminescence imaging. Soybean peroxidase (SBP) was used to label f-PSA or t-PSA monoclonal antibody. With the amplification effects of two enhancers, 3-(10'-phenothiazinyl) propane-1-sulfonate (SPTZ) and 4-morpholinopyridine (MORP), the CL intensity could significantly enhanced, which improved the sensing sensitivity and detection limit. Under the optimal conditions, the linear response to the analyte concentration ranged from 0.01-36.7ng/mL and 0.02-125ng/mL for f-PSA and t-PSA, respectively. The results for the detection of forty serum samples from prostate cancer patients and cancer-free patients showed good agreement with the clinical data, suggesting that the proposed assay had acceptable accuracy. The proposed CL imaging immunoassay possess high throughput and acceptable reproducibility, stability and accuracy, which made it great potential to available to distinguish different isoforms of PSA in serum samples.

Involvement of NFκB signaling in mediating the effects of GRK5 on neural stem cells.

Nuclear factor κB (NFκB) signaling plays ubiquitous roles in inflammation, immune response and neurogenesis. G protein-coupled receptor kinase 5 (GRK5) can protect neurons from degeneration. GRK5 also mediates tumor necrosis factor-α (TNFα)-induced NFκB signaling through the phosphorylation of IκBα. Here, we show that NFκB signaling is involved in neural stem cell (NSC) differentiation. The IκBα/p65 pathway was activated by phorbol myristate acetate (PMA), a stimulator of protein kinase C (PKC). Once the NFκB was activated, the initial stage of neural differentiation was induced, with an increased level of GRK5 in NSCs. This finding was reversed in response to the NFκB inhibitor N-acetyl cysteine (NAC). To evaluate the effect of GRK5-NFκB signaling crosstalk on NSC neurogenesis and apoptosis, GRK5 was knocked down by siRNAs in cell culture. SiRNAs against GRK5 not only impaired neural differentiation and axogenesis, but also induced apoptosis of NSC. GRK5 knockdown affected the transcription of NFκB, phosphorylation of the liver kinase B1 (LKB1) and the activity of caspase 3, thereby modulated neurogenesis and apoptosis. Taken together, our findings reveal a novel function of GRK5 in neurogenesis and provide insight into the molecular mechanisms underlying neurodevelopmental disorders and neurodegenerative diseases.

Construction of photoelectrochemical thrombin aptasensor via assembling multilayer of graphene-CdS nanocomposites.

A photoelectrochemical (PEC) aptasensor for highly sensitive and specific detection of thrombin was developed by using graphene­CdS nanocomposites multilayer as photoactive species and electroactive mediator hexaammineruthenium(III) chloride (Ru(NH(3))(6)(3+)) as signal enhancer. Graphene­CdS nanocomposites (G­CdS) were synthesized by one-pot reduction of oxide graphene and CdCl2 with thioacetamide. The photoactive multilayer was prepared by alternative assembly of the negatively charged 3-mercaptopropionic acid modified graphene­CdS nanocomposites (MPA-G­CdS) and the positively charged polyethylenimine (PEI) on ITO electrode. This layer-by-layer assembly method enhanced the stability and homogeneity of the photocurrent readout of G­CdS. Thrombin aptamer was covalently bound to the multilayer by using glutaraldehyde as cross-linking. Electroactive mediator (Ru(NH(3))(6)(3+)) could interact with the DNA phosphate backbone and thus facilitated the electron transfer between G­CdS multilayer and electrode and enhanced the photocurrent. Hybridizing of a long complementary DNA with thrombin aptamer could increase the adsorption amount of (Ru(NH(3))(6)(3+)), which in turn boosted the signal readout. In the presence of target thrombin, the affinity interaction between thrombin and its aptamer resulted in the long complementary DNA releasing from the G­CdS multilayer and decreasing of photocurrent signal. On the basis of G­CdS multilayer as the photoactive species, (Ru (NH(3))(6)(3+)) as an electroactive mediator, and aptamer as a recognition module, a high sensitive PEC aptasensor for thrombin detection was proposed. The thrombin aptasensor displayed a linear range from 2.0 pM to 600.0 pM and a detection limit of 1.0 pM. The present strategy provided a promising ideology for the future development of PEC biosensor.

Sensitive and antifouling impedimetric aptasensor for the determination of thrombin in undiluted serum sample.

A highly sensitive and attractive antifouling impedimetric aptasensor for the determination of thrombin in undiluted serum sample was developed. The aptasensor was fabricated by co-assembling thiol-modified anti-thrombin binding aptamer, dithiothreitol and mercaptohexanol on the surface of gold electrode. The performance of aptasensor was characterized by atomic force microscopy, contact angle and electrochemical impedance spectroscopy. In the measurement of thrombin, the change in interfacial electron transfer resistance of aptasensor was monitored using a redox couple of Fe(CN)(6)(3-/4-). The increase in the electron transfer resistance was linearly proportional to the concentration of thrombin in the range from 1.0 to 20ng/mL and a detection limit of 0.3ng/mL thrombin was achieved. The fabricated aptasensor displayed attractive antifouling properties and allowed direct quantification of extrinsic thrombin down to 0.08ng/mL in undiluted serum sample. This work provides a promising strategy for clinical application with impressive sensitivity and antifouling characteristics.

Sensitive competitive flow injection chemiluminescence immunoassay for IgG using gold nanoparticle as label.

A sensitive competitive flow injection chemiluminescence (CL-FIA) immunoassay for immunoglobulin G (IgG) was developed using gold nanoparticle as CL label. In the configuration, anti-IgG antibody was immobilized on a glass capillary column surface by 3-(aminopropyl)-triethoxysilane and glutaraldehyde to form immunoaffinity column. Analyte IgG and gold nanoparticle labeled IgG were passed through the immunoaffinity column mounted in a flow system and competed for the surface-confined anti-IgG antibody. CL emission was generated from the reaction between luminol and hydrogen peroxide in the presence of Au (III), generated from chemically oxidative dissolution of gold nanoparticle by an injection of 0.10 mol L(-1) HCl-0.10 mol L(-1) NaCl solution containing 0.10 mmol L(-1) Br(2). The concentration of analyte IgG was inversely related to the amount of bound gold nanoparticle labeled IgG and the CL intensity was linear with the concentration of analyte IgG from 1.0 ng mL(-1) to 40 ng mL(-1) with a detection limit of 5.2×10(-10) gm L(-1). The whole assay time including the injections and washing steps was only 30 min for one sample, which was competitive with CL immunoassays based on a gold nanoparticle label and magnetic separation. This work demonstrates that the CL immunoassay incorporation of nanoparticle label and flow injection is promising for clinical assay with sensitivity and high-speed.