Chinese Herbal Medicine Xueshuantong Enhances Cerebral Blood Flow and Improves Neural Functions in Alzheimer's Disease Mice.

Reduced cerebral blood flow in Alzheimer's disease (AD) may occur in early AD, which contributes to the pathogenesis and/or pathological progression of AD. Reversing this deficit may have therapeutic potential. Certain traditional Chinese herbal medicines (e.g., Saponin and its major component Xueshuantong [XST]) increase blood flow in humans, but whether they could be effective in treating AD patients has not been tested. We found that systemic XST injection elevated cerebral blood flow in APP/PS1 transgenic mice using two-photon time-lapse imaging in the same microvessels before and after injection. Subchronic XST treatment led to improved spatial learning and memory and motor performance in the APP/PS1 mice, suggesting improved neural plasticity and functions. Two-photon time lapse imaging of the same plaques revealed a reduction in plaque size after XST treatment. In addition, western blots experiments showed that XST treatment led to reduced processing of amyloid-ß protein precursor (AßPP) and enhanced clearance of amyloid-ß (Aß) without altering the total level of AßPP. We also found increased synapse density in the immediate vicinity of amyloid plaques, suggesting enhanced synaptic function. We conclude that targeting cerebral blood flow can be an effective strategy in treating AD.

Platelet-derived growth factor receptor beta identifies mesenchymal stem cells with enhanced engraftment to tissue injury and pro-angiogenic property.

Mesenchymal stem cells (MSCs) are heterogeneous likely consisting of subpopulations with various therapeutic potentials. Here we attempted to acquire a subset of MSCs with enhanced effect in wound healing. We found that human placental MSCs expressing platelet-derived growth factor (PDGF) receptor (PDGFR)-ß exhibited greater proliferation rates and generated more colony-forming unit-fibroblast (CFU-F), compared to PDGFR-ß- MSCs. Notably, PDGFR-ß+ MSCs expressed higher levels of pro-angiogenic factors such as Ang1, Ang2, VEGF, bFGF and PDGF. When 106 GFP-expressing MSCs were topically applied into excisional wounds in mice, PDGFR-ß+ MSCs actively incorporated into the wound tissue, resulting in enhanced engraftment (3.92 ± 0.31 × 105 remained in wound by 7 days) and accelerated wound closure; meanwhile, PDGFR-ß- MSCs tended to remain on the top of the wound bed with significantly fewer cells (2.46 ± 0.26 × 105) engrafted into the wound, suggesting enhanced chemotactic migration and engraftment of PDGFR-ß+ MSCs into the wound. Real-Time PCR and immunostain analyses revealed that the expression of PDGF-B was upregulated after wounding; transwell migration assay showed that PDGFR-ß+ MSCs migrated eightfold more than PDGFR-ß- MSCs toward PDGF-BB. Intriguingly, PDGFR-ß+ MSC-treated wounds showed significantly enhanced angiogenesis compared to PDGFR-ß- MSC- or vehicle-treated wounds. Thus, our results indicate that PDGFR-ß identifies a subset of MSCs with enhanced chemotactic migration to wound injury and effect in promoting angiogenesis and wound healing, implying a greater therapeutic potential for certain diseases.

Dissociation of haemolytic and oligomer-preventing activities of gramicidin S derivatives targeting the amyloid-ß N-terminus.

The intrinsic haemolysis of an amyloid-ß (Aß) N-terminal targeting gramicidin S derivative was successfully dissociated from its Aß oligomer-preventing activities via Ala-scanning-based regulation of molecular amphiphilicity. The representative analogue DGR-7 shows low toxicity but significant efficiency in preventing Aß oligomers and reducing amyloid plaques in APP/PS1 transgenic AD mice.

Preparation of pH-Sensitive Dextran Nanoparticle for Doxorubicin Delivery.

One of challenge for cancer therapy is efficient delivery of anticancer agents into tumor sites to increase efficiency of drugs and reduce side effects. To overcome this challenge, we designed pH- sensitive doxorubicin prodrug (DEX-PEI-DOX) nanoparticles based on dextran-poly(ethylene imine) copolymers (DEX-PEI). The DEX-PEI-DOX conjugates were conveniently prepared by grafting PEI to dextran, and then anticancer drug doxorubicin (DOX) were conjugated to DEX-PEI through acid cleavable cis-aconityl bonds. The experiments of dynamic light scattering (DLS) and transmission electron microscopy (TEM) represented that size of dextran nanoparticles was about 120 nm with uniform spherical shape. In vitro drug release from self-assembled nanoparticles was dependent on the pH of medium due to the cis-aconityl linkage. Confocal images revealed that dextran based pH-sensitive DOX delivery nanoparticle could enter into Human breast carcinoma (MCF-7) cells easily. Therapeutic efficacy against MCF-7 cells in vitro was evaluated through MTT assays and the results showed that dextran nanoparticle had obvious anticancer ability. All above results indicated this pH-sensitive DOX-loaded nanoparticles system would be a useful candidate for cancer therapy.

Dextran based sensitive theranostic nanoparticles for near-infrared imaging and photothermal therapy in vitro.

Indocyanine green (ICG) nanoparticles were developed via electrostatic interactions of ICG and dextran based block copolymers (PEG-dextran(-SS-NH2)) as near-infrared (NIR) theranostic nanoparticles. The nanoparticles could be activated from "OFF" to "ON" of NIR fluorescence in an intracellular environment and used for NIR imaging and photothermal therapy.

PEI protected aptamer molecular probes for contrast-enhanced in vivo cancer imaging.

Aptamers have emerged as promising molecular probes for cancer diagnosis. However, their application for in vivo cancer imaging remains limitation due to the poor stability in blood and the degradation by nucleases. In the present study, we generated PEI/aptamer molecular complexes for cancer imaging in vivo by using deoxyribonuclease (DNase)-activatable fluorescence probes (DFProbes) to monitor DNA degradation. The results showed that the complexes with PEI at the N/P ratio from 3.8 to 15 effectively prevented the degradation of DFProbes both in vitro and in vivo. Moreover, PEI successfully protected TD05 aptamers from DNase degradation without affecting its specific recognition of Ramos cells. In tumor bearing mice, PEI/aptamer molecular complexes further demonstrated superior passive tumor targeting and extended circulation time as compared with free aptamer. Hence, the well-defined PEI/aptamer probe is a novel strategy to deliver targeted aptamer for tumor diagnosis and imaging in vivo.

Indocyanine green-loaded biodegradable tumor targeting nanoprobes for in vitro and in vivo imaging.

Indocyanine green (ICG) is a near-infrared (NIR) fluorescence dye for extensive biological application, but limited by its poor aqueous stability in vitro, concentration-dependent aggregation, rapid elimination from the body, and lack of target specificity. In this paper, to overcome these limitations, folate receptor-targeted, ICG dye-doped poly(d,l-lactide-co-glycolide) (PLGA) lipid nanoparticles (FA-ICG-PLGA-lipid NPs) were constructed by a single-step self-assemble and nanoprecipitation method. The prepared FA-ICG-PLGA-lipid NPs exhibited good biocompatibility, monodispersity, excellent NIR penetration ability, significant stability against photobleaching and long circulation time. The intracellular uptake experiment proved the targeting efficacy of the FA-ICG-PLGA-lipid NPs was more effective in folate receptor over-expressing MCF-7 cells than folate receptor negative A549 cells. Furthermore, the in vivo experiments showed the FA-ICG-PLGA-lipid NPs were specifically targeted to the tumor, and its circulation time was much longer than free ICG. These biocompatible and biodegradable NIR-NPs prove a potential application in tumor diagnosis and targeted imaging due to its high aqueous stability, excellent NIR optical properties and significantly targeting property in vivo.

DNase-activatable fluorescence probes visualizing the degradation of exogenous DNA in living cells.

This work presents a method to visualize the degradation of exogenous DNA in living cells using a novel type of activatable fluorescence imaging probe. Deoxyribonuclease (DNase)-activatable fluorescence probes (DFProbes) are composed of double strands deoxyribonucleic acid (dsDNA) which is labeled with fluorophore (ROX or Cy3) and quencher on the end of one of its strands, and stained with SYBR Green I. In the absence of DNase, DFProbes produce the green fluorescence signal of SYBR Green I. In the presence of DNase, SYBR Green I is removed from the DFProbes and the labeled fluorophore is separated from the quencher owing to the degradation of DFProbes by DNase, resulting in the decrease of the green fluorescence signal and the occurrence of a red fluorescence signal due to fluorescence resonance energy transfer (FRET). DNase in biological samples was detected using DFProbes and the fluorescence imaging in living cells was performed using DFprobe-modified Au nanoparticles. The results show that DFProbes have good responses to DNase, and can clearly visualize the degradation of exogenous DNA in cells in real time. The well-designed probes might be useful in tracing the dynamic changes of exogenous DNA and nanocarriers in vitro and in vivo.

A universal quantum dots-aptamer probe for efficient cancer detection and targeted imaging.

Targeted quantum dots have shown as an analytical and imaging tool for cancer detection and molecular imaging. Aptamers have recently been demonstrated as ideal candidates for molecular targeting applications. In the present work, quantum dots (QDs) were encapsulated with functional poly(ethylene glycol)-phospholipids to improve their solubility in water solution. The as-prepared QDs were then conjugated with the AS1411 aptamer recognizing the nucleolin overexpressed on cancer cell surface. The recognition of QD-Aptamer nanocomplex to breast cancer cells was demonstrated using confocal microscopy, and the viability of QDs-aptamer bioconjugate bound cells were not affected within 24 h, indicating that the probe was biocompatible and suitable for in vitro diagnostic assays and live cell imaging. Such well-defined aptamer-conjugated QDs nanoprobe was simple and universal to be extended to prepare various aptamer-nanoparticles hybrid systems for cancer targeting and molecular imaging applications.

Methylene blue-encapsulated phosphonate-terminated silica nanoparticles for simultaneous in vivo imaging and photodynamic therapy.

A bifunctional nanoparticles-based carrier for simultaneous in vivo imaging and photodynamic therapy by encapsulating methylene blue (MB) alone in the phosphonate-terminated silica matrix has been developed. The phosphonate-terminated silica nanoparticles, entrapping water-soluble photosensitizer MB (MB-encapsulated PSiNPs), are synthesized by the controlled synchronous hydrolysis of tetraethoxysilane and trihydroxyl silyl propyl methyl phosphonate in the water-in-oil microemulsion. The resulting MB-encapsulated PSiNPs effectively prevent the leakage of entrapped MB from the particles and provide protection for against reduction by diaphorase. Enough dose of irradiation to the MB-encapsulated PSiNPs under the light of 635 nm results in efficient generation of singlet oxygen and induces photodynamic damage to Hela cells. Furthermore, the non-invasive visualization of MB-encapsulated PSiNPs in mice under the in vivo imaging system confirmed the MB-encapsulated PSiNPs also presents near-infrared luminescence for in vivo imaging. And the effect of the PDT toward the xenograft tumor in vivo is exciting after imaging the MB-encapsulated PSiNPs injected tumor using in vivo optical imaging system. Thus, the single particle platform is effective for simultaneous in vivo imaging and photodynamic therapy without using extra agent, which can provide image-guidance for site-specific photodynamic therapy.