Aiqing Li

Nanoplated bismuth titanate sub-microspheres for protein immobilization and their corresponding direct electrochemistry and electrocatalysis

A layered inorganic perovskite sub-micrometer-scale material, nanoplated bismuth titanate (Bi(4)Ti(3)O(12)) sub-microspheres (NBTSMs) constructed with tens of Bi(4)Ti(3)O(12) nanoplates, was for the first time synthesized by a facile hydrothermal synthesis strategy. The NBTSMs were employed as a supporting matrix to explore a novel immobilization and biosensing platform of redox proteins through a combined hydrogen bond and electrostatic assembly process. Biocompatibility, stability, reproducibility, and electrochemical and electrocatalytic properties of the resulting NBTSMs-based composite were studied by UV-vis absorption, FTIR, and electrochemical methods. The research results revealed that the NBTSMs-based composite was a satisfying matrix for proteins to effectively retain their native structure and bioactivity. With advantages of the Bi(4)Ti(3)O(12) layered material, facilitated direct electron transfer of the metalloenzymes with an apparent heterogeneous electron transfer rate constant (k(s)) of 20.0+/-3.8s(-1) was acquired on the NBTSMs-based enzyme electrode. The NBTSMs-based biosensor demonstrated significant electrocatalytic activity for the reduction of hydrogen peroxide with an apparent Michaelis-Menten constant (204 microM), wide linear range (2-430 microM), and low detection limit (0.46 microM, S/N=3). These indicated that the nanoplate-constructed Bi(4)Ti(3)O(12) sub-microspheres were one of ideal candidate materials for direct electrochemistry of redox proteins and the construction of the related enzyme biosensors, and may find potential applications in biomedical, food, and environmental analysis and detection.

Sol–gel hydrothermal synthesis and enhanced biosensing properties of nanoplated lanthanum-substituted bismuth titanate microspheres

Nanoplated lanthanum-substituted bismuth titanate (Bi3.25La0.75Ti3O12, BLTO) microspheres constructed with tens of BLTO nanoplates were synthesized by a sol–gel hydrothermal method. Using nanoplated BLTO microspheres, a novel third-generation H2O2 biosensor was fabricated with loading of myoglobin (Mb) and chitosan (Chi). Scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and X-ray diffraction (XRD) measurements reveal that partial Bi ions of bismuth titanate (BTO, Bi4Ti3O12) are successfully substituted with La ions by the sol–gel hydrothermal method. UV-visible (UV-vis) and Fourier-transform infrared (FTIR) spectra show that Mb encapsulated in the Mb–Chi–BLTO film can retain its bioactivity well. Comparative experiments witness that the Mb–Chi–BLTO biosensor, compared with the Mb–Chi–BTO biosensor, not only has enhanced direct electron-transfer capacity (e.g., stronger redox peak currents (approximately 3-fold) and a larger heterogeneous electron-transfer rate constant of 12.8 ± 3.3 s−1), but also exhibits a wider linear response to H2O2 in the concentration range of 2–490 μM, higher sensitivity (88 mA cm−2 M−1), a lower Michaelis–Menten constant (0.55 mM) and detection limit (0.14 μM), a shorter response time (2.8 s), and better reproducibility and stability. These results imply that La doping greatly improves electrochemical and electrocatalytic properties of the Mb–Chi–BLTO biosensor, which will open up a new idea for the design of third-generation electrochemical biosensors, and the BLTO-based biosensors are also expected to find potential applications in many areas such as clinical diagnosis and food and environmental detection.

EDTA-directed self-assembly and enhanced catalytic properties of sphere-constructed platinum nanochains

A simple and synthetically directed self-assembly approach to the construction of platinum nanochains (PtNCs) has been demonstrated. Micrometre-length PtNCs constructed with spherical Pt nanoparticles (PtNPs) of about 5 nm were synthesized by the reduction of Pt–EDTA (ethylenediaminetetraacetic acid) chelate complex with sodium borohydride (NaBH4). It was found that EDTA played a critical role in the formation of sphere-constructed PtNCs. The PtNPs–Nafion/glassy carbon (GC) and PtNCs–Nafion/GC modified electrodes were fabricated and their corresponding electrocatalytic activities were studied. Comparative studies demonstrated that the PtNCs catalyst had a higher electrochemically active surface area (ECSA, almost 2 times) and better catalytic activity (almost 10 times) in comparison with the PtNPs catalyst. The PtNCs catalyst with a high ECSA and catalyst activity will be a good anode catalyst candidate for direct methanol fuel cells.

Al3+-directed self-assembly and their electrochemistry properties of three-dimensional dendriform horseradish peroxidase/polyacrylamide/platinum/single-walled carbon nanotube composite film.

A novel general methodology for protein immobilization and third-generation biosensor construction is demonstrated, which involves Al(3+)-directed polyacrylamide (PAM) self-assembly into an ordered dendriform structure, easily immobilizing enzymes and nanoparticles. Platinum/single-walled carbon nanotube (Pt/SWCNT) heterojunction nanomaterials were for the first time fabricated via an EDTA-directed synthesis strategy. The Pt/SWCNTs were employed as a supporting matrix to explore a novel immobilization and biosensing platform of redox proteins through cooperating Al(3+)-directed PAM self-assembly. Compared with the almost single-layer horseradish peroxidase (HRP)/PAM film electrode, multilayer HRP/PAM/Pt/SWCNT film electrode exhibited a pair of much stronger redox peaks at -0.22 V (vs. Ag/AgCl). Moreover, with advantages of the ordered multilayer HRP/PAM/Pt/SWCNT film, facilitated direct electron transfer of the metalloenzymes with an apparent heterogeneous electron transfer rate constant (k(s)) of 14.94+/-1.36 s(-1) and smaller peak-to-peak separation (DeltaE(p)) of about 37 mV was acquired on the PAM/Pt/SWCNT-based enzyme electrode. The PAM/Pt/SWCNT-based biosensor demonstrated significant electrocatalytic activity for the reduction of hydrogen peroxide with a small apparent Michaelis-Menten constant (87 microM), wide linear range (1-270 microM), very low detection limit (0.08 microM, S/N=3), and high sensitivity (372 mA cm(-2) M(-1)). Together, these indicated that the Al(3+)-directed HRP/PAM/Pt/SWCNT film was one of ideal candidate materials for direct electrochemistry of redox proteins and the construction of the related enzyme biosensors, and may find potential applications in biomedical, food, and environmental analysis and detection.

Coordinatively Self-Assembled Luminescent Gold Nanoparticles: Fluorescence Turn-On System for High-Efficiency Passive Tumor Imaging

A fluorescence turn-on system for highly efficient and prolonged tumor imaging has been established by a Co2+-induced coordination self-assembly strategy, in which luminescent glutathione (GSH)-modified gold nanoparticles (LGAuNPs) are assembled into LGAuNPs assemblies (LGAuNPs-Co) through a coordination bond between an unoccupied orbit of Co2+ and lone pair electrons of GSH on the surface of LGAuNPs. The LGAuNPs-Co is sensitive to microenvironment pH, and its quenched luminescence will be turned on in tumor tissues (acidic microenvironment), which behaves as a fluorescence turn-on system for passive tumor imaging. The fluorescence turn-on system combines advantages of the enhanced permeability and retention (EPR) effect of NPs and pH-induced fluorescence turn-on property at the tumor site, which results in a larger fluorescence intensity (FI) difference between normal and tumor tissues as compared with that of luminescent Au NPs (LAuNPs, only with the EPR effect) (∼12-fold). Such a large FI difference results in that LGAuNPs-Co has rapid (∼1.6 h), persistent (∼24 h p.i.), and highly efficient tumor targeting capability in comparison with LGAuNPs. Moreover, the LGAuNPs-Co also has much longer tumor retention, faster renal clearance, and lower reticuloendothelial system (RES) uptake than LGAuNPs. Therefore, the fluorescence turn-on system is very promising for cancer diagnosis and therapy.

Increased Apoptosis in the Paraventricular Nucleus Mediated by AT1R/Ras/ERK1/2 Signaling Results in Sympathetic Hyperactivity and Renovascular Hypertension in Rats after Kidney Injury

Background: The central nervous system plays a vital role in the development of hypertension, but the molecular regulatory mechanisms are not fully understood. This study aimed to explore signaling in the paraventricular nucleus (PVN) which might contribute to renal hypertension. Methods: Renal hypertension model was established by five-sixth nephrectomy operation (5/6Nx) in male Sprague Dawley rats. Ten weeks afterwards, they were random assigned to no treatment, or intracerebroventricular injection (ICV) with artificial cerebrospinal fluid, losartan [angiotensin II receptor type 1 (AT1R) antagonist], farnesylthiosalicylic acid (Ras inhibitor), PD98059 (MEK inhibitor), or SB203580 (p38 inhibitor) and Z-DEVD-FMK (caspase-3 inhibitor). Before and after treatment, physiological and biochemical indices were measured. Immunohistochemistry, western blot and RT-PCR were applied to quantify key components of renin-angiotensin system, apoptosis-related proteins, Ras-GTP, and MAPKs in the PVN samples. TUNEL assay was used to measure the situ apoptosis in PVN. Results: The 5/6Nx rats showed significantly elevated systolic blood pressure, urinary protein excretion, serum creatinine, and plasma norepinephrine (p < 0.05) compared to sham rats. The expression of angiotensinogen, Ang II, AT1R, p-ERK1/2, or apoptosis-promoting protein Bax were 1.08-, 2.10-, 0.74-, 0.82-, 0.83-fold higher in the PVN of 5/6Nx rats, than that of sham rats, as indicated by immunohistochemistry. Western blot confirmed the increased levels of AT1R, p-ERK1/2 and Bax; meanwhile, Ras-GTP and p-p38 were also found higher in the PVN of 5/6Nx rats, as well as the apoptosis marker cleaved caspase-3 and TUNEL staining. In 5/6Nx rats, ICV infusion of AT1R antagonist, Ras inhibitor, MEK inhibitor or caspase-3 inhibitor could lower systolic blood pressure (20.8-, 20.8-, 18.9-, 14.3%-fold) together with plasma norepinephrine (53.9-, 57.8-,63.3-, 52.3%-fold). Western blot revealed that blocking the signaling of AT1R, Ras, or MEK/ERK1/2 would significantly reduce PVN apoptosis as indicated by changes of apoptosis-related proteins (p < 0.05). AT1R inhibition would cause reduction in Ras-GTP and p-ERK1/2, but not vice versa; such intervention with corresponding inhibitors also suggested the unidirectional regulation of Ras to ERK1/2. Conclusion: These findings demonstrated that the activation of renin-angiotensin system in PVN could induce apoptosis through Ras/ERK1/2 pathway, which then led to increased sympathetic nerve activity and renal hypertension in 5/6Nx rats.

Hydrosoluble 50% N-acetylation-thiolated chitosan complex with cobalt as a pH-responsive renal fibrosis targeting drugs

Abstract About 50% N-acetylation-thiolated chitosan possessing good water solubility was modified from commercial low-molecular-weight chitosan. Chitosan performed obvious target toward renal tubular epithelial cells, and bivalent cobalt ions improved the renal fibrosis inflammation significantly. There were many complexation sites on chitosan after being modified with sulfydryl. So sulfydryl played a role of connecting bridge between chitosan and cobalt ions. Then, this N-acetylation-thiolated chitosan cobalt (NTCC) nanocomplex was designed. The nanocomplex showed excellent stability under normal physiological conditions, and cobalt would be released from the biomaterials in acidic environment. As it was affected by inflammation, the pH in renal fibrosis lesion region was acidic. So there was a specific drug release process happening in lesion region. And drug release efficiency was determined by acidity, which demonstrated that lower the acidity, the faster and more the cobalt ion release. When this nanocomplex was intraperitoneally injected into ureter-obstructed mice, obvious attenuation of fibrotic progression was shown. It was demonstrated that NTCC exhibited special renal-targeting capacity and could be chosen as drug for treating renal fibrosis.

Advanced oxidation protein products promote NADPH oxidase-dependent β-cell destruction and dysfunction through the Bcl-2/Bax apoptotic pathway.

The accumulation of plasma advanced oxidation protein products (AOPPs) has been linked with diverse disorders, including diabetes, chronic kidney disease, obesity, and metabolic syndrome. The aim of the present study was to evaluate the pathophysiological relevance of AOPPs in β-cell destruction and dysfunction. Exposure of cultured rat β-cells (INS-1) to AOPPs induced an increase in Bax expression, caspase-3 activity, and apoptosis as well as a decrease in Bcl-2 expression in a dose- and time-dependent manner. AOPP challenge rapidly increased the production of intracellular superoxide by activation of NADPH oxidases, demonstrated by p47phox translocation and interaction with p22phox and gp91phox, and this in turn led to apoptosis. AOPPs treatment resulted in β-cell apoptosis, AOPPs accumulation, and decreased insulin content in pancreas and plasma in unilateral nephrectomized rats. Chronic inhibition of NADPH oxidase by apocynin prevented β-cell apoptosis and ameliorated insulin deficiency in AOPP-challenged rats. This study demonstrates for the first time that accumulation of AOPPs promotes NADPH oxidase-dependent β-cell destruction and dysfunction by the Bcl-2/Bax-caspase apoptotic pathway. This finding may provide a mechanistic explanation for β-cell destruction and dysfunction in patients with diverse disorders.

Inlaid Nd-substituted bismuth titanate nanoplates for protein immobilization and Nd-controlled electrochemical properties

Neodymium (Nd) substituted bismuth titanate (Bi(4-x)Nd(x)Ti(3)O(12), BNTO-x) nanoplates inlaid one another were prepared by sol-gel hydrothermal method, which was explored for protein immobilization and biosensor fabrication. Comparative experiments witnessed that Bi(3+) ions in bismuth titanate (Bi(4)Ti(3)O(12), BTO) were successfully substituted with Nd(3+) ions, and the electrochemical properties of the Hb-Chi-BNTO biosensors closely depended on the Nd(3+) ion content. With increasing the Nd(3+) doping content, the electrochemical performance of the Hb-Chi-BNTO-x biosensors showed regularly variable. Moreover, compared with the Hb-Chi-BTO and other Hb-Chi-BNTO-x biosensors, the Hb-Chi-BNTO-0.85 biosensor had more excellent electrochemical and electrocatalytic properties such as stronger redox peak currents (approximately three-fold), smaller peak-to-peak separation (50 mV), larger heterogeneous electron transfer rate (14.1 ± 3.8s(-1)), higher surface concentration of electroactive redox protein (about 8.16 × 10(-11)mol/cm(2)), and better reproducibility and stability. The Nd-depended electrochemical properties of the Hb-Chi-BNTO biosensors may open up a new idea for designing third-generation electrochemical biosensors, and the BNTO-0.85-based biosensor is also expected to find potential applications in many areas such as biomedical, food, and environmental detection.

Highly Stabilized Core-Satellite Gold Nanoassemblies in Vivo: DNA-Directed Self-Assembly, PEG Modification and Cell Imaging

Au nanoparticles (NPs) have important applications in bioimaging, clinical diagnosis and even therapy due to its water-solubility, easy modification and drug-loaded capability, however, easy aggregation of Au NPs in normal saline and serum greatly limits its applications. In this work, highly stabilized core-satellite Au nanoassemblies (CSAuNAs) were constructed by a hierarchical DNA-directed self-assembly strategy, in which satellite Au NPs number could be effectively tuned through varying the ratios of core-AuNPs-ssDNA and satellite-AuNPs-ssDNAc. It was especially interesting that PEG-functionalized CSAuNAs (PEG-CSAuNAs) could not only bear saline solution but also resist the enzymatic degradation in fetal calf serum. Moreover, cell targeting and imaging indicated that the PEG-CSAuNAs had promising biotargeting and bioimaging capability. Finally, fluorescence imaging in vivo revealed that PEG-CSAuNAs modified with N-acetylation chitosan (CSNA) could be selectively accumulate in the kidneys with satisfactory renal retention capability. Therefore, the highly stabilized PEG-CSAuNAs open a new avenue for its applications in vivo.