ABSTRACT
Carotenoids play a pivotal role in plant. Tagetes erecta, commonly called marigold, has increasing nutritional and economic value due to its high level of carotenoids in flower. However, the functional genes in the carotenoid biosynthesis of T. erecta have not been studied. In this work, three T. erecta varieties with flowers of yellow, yellow-orange and orange color, respectively, were examined for carotenoids composition and corresponding expression profiling of biosynthetic genes at four developmental stages. The results indicated that the varieties with higher lutein content, orange-flower 'Juwang' and yellow-orange 'Taishan', exhibited significant upregulation of genes in the upstream biosynthesis pathway, especially PDS (phytoene desaturase), PSY (phytoene synthase) and ZDS (zeta-carotene desaturase), whereas downstream carotenoid cleavage genes CCD (carotenoid cleavage dioxygenase) were markedly downregulated throughout flower development in the highest lutein containing variety 'Juwang'. Furthermore, marigold TePDS, TePSYS3 and TeZDS were isolated and transformed into tomato. Overexpression of TePDS or TeZDS resulted in the promotion of fruit ripening and accumulation of carotenoids in the transgenic lines. On the other hand, marigold TePSYS3 showed multiple effects, not only on fruit carotenogenesis but also on pigmentation patterns in vegetative tissues and plant growth. Taken together, the variations in expression profiles of the biosynthetic genes contribute to dynamic change in carotenoid levels and diversity of flower coloration in T. erecta. These functional genes of T. erecta were verified in tomato and provide targets for genetic improvement of fruit carotenoids accumulation.
Subject(s)
Carotenoids , Flowers , Fruit , Pigmentation , Solanum lycopersicum , Tagetes , Tagetes/metabolism , Tagetes/genetics , Carotenoids/metabolism , Solanum lycopersicum/genetics , Solanum lycopersicum/metabolism , Solanum lycopersicum/growth & development , Flowers/genetics , Flowers/metabolism , Flowers/growth & development , Fruit/genetics , Fruit/metabolism , Fruit/growth & development , Pigmentation/genetics , Gene Expression Regulation, Plant , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/geneticsABSTRACT
Northern Africa has become the first region in the world to exhaust its water resources, with a 40 % decrease in per capita water availability south of the Sahara over the past decade. While adjusting production structures and consumption can regulate the supply-demand dynamics of water resources, the extent of the impact of virtual water-induced pressure on both the regional and national levels in Africa remains largely understudied. Applying the standard Penman formula, this research calculates the water footprint of eight cereal crops in 54 African countries from 1990 to 2021. By integrating corresponding data on cereal trade, the study analyzes changes in virtual water stress. The findings indicate a decline in the per-unit production and consumption water footprints for African cereals. However, the continuous expansion of cultivation areas contributes to a rising water stress. In comparison to 1990, the water stress for soybeans, sorghum, rice, maize, and cassava increased by 149.72 %, 146.88 %, 133.89 %, 123.30 %, and 90.8 %, respectively, in 2021. Only barley showed a reduction in water stress by 23.22 %. The study underscores the growing interconnectedness of virtual water trade (VWT) among African nations from 1990 to 2021, leading to a more balanced trade distribution. VWT has reduced water stress by 7.65 %, 2.08 %, and 1.8 % in Western, Central, and Northern Africa, respectively, while increasing pressure in Southern and Eastern Africa by 10.51 % and 1.01 %. The flow of virtual water in Africa is most influenced by spatial proximity, primarily occurring between adjacent countries or regions. Forecasts for water stress under the five scenarios of SSPs-RCP8.5 have been conducted, revealing a continuous increase in water stress across Africa. Furthermore, analysis of the SSP2-RCP8.5 scenario indicates that by 2030 and 2040, African cereal crops are projected to face virtual water resource stress increases of 7 % and 18.76 %, respectively, compared to 2020 levels. During the same period, Sierra Leone is anticipated to experience a growth rate in virtual water stress of approximately 1903.38 %. Consequently, altering crop cultivation structures and enhancing VWT are poised to alleviate water resource pressure, promoting the scientific management of agricultural water resources in Africa.
ABSTRACT
We conducted this study to investigate the radioprotective effects of recombinant human thrombopoietin (rhTPO) on beagle dogs irradiated with 3.0 Gy 60Co gamma rays. Fifteen healthy adult beagles were randomly assigned to a control group with alleviating care, and 5 and 10 µg/kg rhTPO treatment group. All animals received total-body irradiation using 60Co γ-ray source at a dose of 3.0 Gy (dose rate was 69.1 cGy/min). The treatment group received intramuscular injection of rhTPO 5 and 10 µg/kg at 2 h postirradiation, and the control group was administrated the same volume of normal saline. The survival rate, clinical signs, peripheral hemogram, serum biochemistry, and histopathological examination of animals in each group were assessed. Single administration of 10 µg/kg rhTPO at 2 h postirradiation promoted the recovery of multilineage hematopoiesis and improved the survival rate of beagles irradiated with 3 Gy 60Co γ rays. The administration of 10 µg/kg rhTPO alleviated fever and bleeding, reduced the requirement for supportive care, and may have mitigated multiple organ damage.
Subject(s)
Gamma Rays , Hematopoiesis , Radiation-Protective Agents , Recombinant Proteins , Thrombopoietin , Whole-Body Irradiation , Animals , Dogs , Thrombopoietin/pharmacology , Thrombopoietin/administration & dosage , Recombinant Proteins/administration & dosage , Recombinant Proteins/pharmacology , Hematopoiesis/drug effects , Hematopoiesis/radiation effects , Humans , Radiation-Protective Agents/pharmacology , Radiation-Protective Agents/administration & dosage , Male , Cobalt Radioisotopes , Female , Dose-Response Relationship, RadiationABSTRACT
OBJECTIVE: To confirm the therapeutic effect of recombinant human thrombopoietin (rhTPO) on rhesus monkeys irradiated with 5.0 Gy 60Co γ-ray, and provide experimental basis for clinical treatment of similar patients. METHODS: Fourteen adult rhesus monkeys were irradiated with 60Co γ-ray on both sides at the dose of 5.0 Gy (dose rate 69.2 cGy/min) to establish the acute radiation sickness model. The monkeys were divided into irradiation group (n=5), rhTPO 5 µg/kg group (n=4) and rhTPO 10 µg/kg group (n=5). Two hours after irradiation, the three groups of monkeys were injected with saline 0.1 ml/kg, rhTPO 5 µg/kgï¼0.1 ml/kgï¼ and rhTPO 10 µg/kg(0.2 ml/kg), respectively. The general signs, survival, peripheral hemogram and serum biochemistry of rhesus monkeys were observed before and after irradiation, and the differences between rhTPO group and irradiation control group were compared. RESULTS: After total body irradiation with 5.0 Gy60Co γ-ray, rhesus monkeys successively showed fever, hemorrhage, sharp decrease of whole blood cell counts in peripheral blood and disorder of serum biochemical indexes. Compared with the irradiated control group, a single intramuscular injection of rhTPO 5 µg/kg or 10 µg/kg 2 hours after irradiation could improve the symptoms of fever and bleeding, increase the nadir of peripheral red blood cells and platelets counts, shorten the duration of hemocytopenia, and advance the time for blood cells to return to the pre-irradiation level. The serum biochemical results showed that rhTPO could improve the abnormality of serum biochemical indexes in rhesus monkeys induced by 5.0 Gy total body irradiation to some extent. Compared with the two administration groups, the therapeutic effect of rhTPO 10 µg/ kg was better. CONCLUSION: A single injection of rhTPO 5 µg/ kg or 10 µg/ kg 2 hours after irradiation can alleviate the injury of multilineage hematopoiesis and promote the recovery in monkeys irradiated by 5.0 Gy γ-ray. It also improves animal signs and has obvious therapeutic effect on acute radiation sickness.
Subject(s)
Radiation Injuries , Humans , Animals , Macaca mulattaABSTRACT
Eyebox uniformity is an important indicator to evaluate the performance of optical waveguide displays. However, there is currently no standard design approach that achieves ideal uniformity over the full field of view (FOV) within the eyebox. Here, a novel method for eyebox uniformity optimization based on linked list processing is proposed. The linked list processing method can fast record the light trajectory and calculate the optimal numerical diffraction efficiency distribution of the coupler. We use the linked list method for an L-shaped diffractive optical waveguide and solve the matched coupler structure by combining rigorous coupled-wave analysis (RCWA) and the simplex method. By building the model on LightTools and demonstrating the illuminance uniformity, the feasibility of the method is verified. In the FOV range of 15°× 15°, the eyebox uniformity reaches 0.9 at the central viewing angle and the overall eyebox uniformity is 0.617.
ABSTRACT
In the post-COVID era, how to improve the level of regional sustainable development has attracted much attention. And the vigorous development of the sports economy may be closely related to the regional sustainable development. This paper explores the impact and mechanism analysis of government sports public expenditure on regional sustainable development from the perspective of sports economic development. The study found that China's sustainable development presents obvious ladder-like characteristics and highlights the regional imbalance and inadequacy of regional green and coordinated development. And the government's increase in public expenditure on sports can significantly promote regional sustainable development and improve the level of regional green and coordinated development. With the continuous improvement of the regional economic development, the effect of sports public expenditure continues to increase. It can be seen from this that implementing the strategy of strengthening the country through sports under the government's guidance is an essential guarantee for the public health and quality of life and the sustainable development of the economy and society. Additionally, the development level of market finance is also an important driving factor for the government's public expenditure on sports to improve the level of sustainable development in the region. From the mechanism analysis, the government activates the local residents' consumption level by increasing the public expenditure on sports, thus promoting regional sustainable development.
Subject(s)
COVID-19 , Quality of Life , COVID-19/epidemiology , COVID-19/prevention & control , China , Economic Development , Humans , Public ExpendituresABSTRACT
BACKGROUND: Rapid diagnosis of infectious pathogens at an early stage is crucial to stabilize the patient's condition, reduce medical costs, and shorten hospital stays. Currently, some point-of-care tests have their own shortcomings. Therefore, we built a microfluidic chip based on loop-mediated isothermal amplification to can quickly and sensitively detect infectious pathogens. METHODS: We extracted the DNA of S. aureus, MRSA, Shigella and Klebsiella pneumoniae. Then, the DNA samples were diluted by 10-fold and examined by two methods: LAMP-microfluidic chip and q-PCR, the sensitivity of whom was also compared. In addition, the specificity of the two was also examined by detecting the target bacteria and other microorganisms using the same methods. Finally, we extracted and tested the DNA of clinically infected humoral samples to determine the coincidence rate between the two methods and the bacterial culture method. RESULTS: For S. aureus, MRSA, Shigella, and Klebsiella pneumoniae, the detection limits of the chip were 2.25 × 103 copies/µl, 5.32 × 103 copies/µl, 2.89 × 103 copies/µl, 6.53 × 102 copies/µl, and the detection limits of q-PCR were 2.25 × 102 copies/µl, 5.32 × 101 copies/µl, 2.89 × 102 copies/µl, 6.53 × 101 copies/µl, respectively. In terms of detection specificity, neither method cross-reacted with other strains. For the detection of infectious humoral samples, the total coincidence rate between the q-PCR and bacterial culture method was 85.7%, 95%, 95%, and 95.5%, and the total coincidence rate between the chip and bacterial culture method was 81%, 95%, 90%, and 86.4%, respectively. CONCLUSION: LAMP-microfluidic chip provides a simple, sensitive, specific, convenient, and rapid pathogen detection method for clinically infected humoral samples without relying on expensive equipment or technical personnels.
Subject(s)
Microfluidics , Staphylococcus aureus , Bacteria/genetics , DNA , Humans , Microfluidics/methods , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques/methods , Polymerase Chain Reaction , Sensitivity and Specificity , Staphylococcus aureus/geneticsABSTRACT
Fluorescently labeled nanoparticles are widely used for evaluating their distribution in the biological environment. However, dye leakage can lead to misinterpretations of the nanoparticles' biodistribution. To better understand the interactions of dyes and nanoparticles and their biological environment, we explored PLGA nanoparticles labeled with four widely used dyes encapsulated (coumarin 6, rhodamine 123, DiI) or bound covalently to the polymer (Cy5.5.). The DiI label was stable in both aqueous and lipophilic environments, whereas the quick release of coumarin 6 was observed in model media containing albumin (42%) or liposomes (62%), which could be explained by the different affinity of these dyes to the polymer and lipophilic structures and which we also confirmed by computational modeling (log PDPPC/PLGA: DiI-2.3, Cou6-0.7). The importance of these factors was demonstrated by in vivo neuroimaging (ICON) of the rat retina using double-labeled Cy5.5/Cou6-nanoparticles: encapsulated Cou6 quickly leaked into the tissue, whereas the stably bound Cy.5.5 label remained associated with the vessels. This observation is a good example of the possible misinterpretation of imaging results because the coumarin 6 distribution creates the impression that nanoparticles effectively crossed the blood-retina barrier, whereas in fact no signal from the core material was found beyond the blood vessels.
ABSTRACT
During the drug development process, many pharmacologically active compounds are discarded because of poor water solubility, but nanoparticle-based formulations are increasingly proposed as a solution for this problem. We therefore studied the distribution of nanoparticulate carriers and the delivery of their poorly water-soluble cargo to a structure of the central nervous system, the retina, under naive and pathological conditions. The lipophilic fluorescent dye coumarin 6 (Cou6) was encapsulated into poly(lactic-co-glycolic acid) PLGA nanoparticles (NPs). After intravenous administration in rats, we analyzed the distribution of cargo Cou6 and of the NP carrier covalently labeled with Cy5.5 in healthy animals and animals with optic nerve crush (ONC). In vivo real-time retina imaging revealed that Cou6 was rapidly released from PLGA NPs and penetrated the inner blood-retina barrier (BRB) within 15 min and PLGA NPs were gradually eliminated from the retinal blood circulation. Ex vivo microscopy of retinal flat mounts indicated that the Cou6 accumulated predominantly in the extracellular space and to a lesser extent in neurons. While the distribution of Cou6 in healthy animals and post ONC was comparable at early time point post-operation, the elimination of the NPs from the vessels was faster on day 7 post ONC. These results demonstrate the importance of considering different kinetics of nano-carrier and poorly water-soluble cargo, emphasizing the critical role of their parenchymal distribution, i.e. cellular/extracellular, and function of different physiological and pathological conditions.
Subject(s)
Nanoparticles , Polyglycolic Acid , Animals , Drug Carriers , Drug Delivery Systems , Lactic Acid , Polylactic Acid-Polyglycolic Acid Copolymer , Rats , Retina , WaterABSTRACT
Hearing deficits impact on the communication with the external world and severely compromise perception of the surrounding. Deafness can be caused by particular mutations in the neuroplastin (Nptn) gene, which encodes a transmembrane recognition molecule of the immunoglobulin (Ig) superfamily and plasma membrane Calcium ATPase (PMCA) accessory subunit. This study investigates whether the complete absence of neuroplastin or the loss of neuroplastin in the adult after normal development lead to hearing impairment in mice analyzed by behavioral, electrophysiological, and in vivo imaging measurements. Auditory brainstem recordings from adult neuroplastin-deficient mice (Nptn-/-) show that these mice are deaf. With age, hair cells and spiral ganglion cells degenerate in Nptn-/- mice. Adult Nptn-/- mice fail to behaviorally respond to white noise and show reduced baseline blood flow in the auditory cortex (AC) as revealed by single-photon emission computed tomography (SPECT). In adult Nptn-/- mice, tone-evoked cortical activity was not detectable within the primary auditory field (A1) of the AC, although we observed non-persistent tone-like evoked activities in electrophysiological recordings of some young Nptn-/- mice. Conditional ablation of neuroplastin in Nptnlox/loxEmx1Cre mice reveals that behavioral responses to simple tones or white noise do not require neuroplastin expression by central glutamatergic neurons. Loss of neuroplastin from hair cells in adult NptnΔlox/loxPrCreERT mice after normal development is correlated with increased hearing thresholds and only high prepulse intensities result in effective prepulse inhibition (PPI) of the startle response. Furthermore, we show that neuroplastin is required for the expression of PMCA 2 in outer hair cells. This suggests that altered Ca2+ homeostasis underlies the observed hearing impairments and leads to hair cell degeneration. Our results underline the importance of neuroplastin for the development and the maintenance of the auditory system.
Subject(s)
Hearing , Animals , Auditory Threshold , Evoked Potentials, Auditory, Brain Stem , Hearing Loss , Membrane Glycoproteins/metabolism , Mice , Mice, Inbred C57BL , Mice, Knockout , Plasma Membrane Calcium-Transporting ATPases/metabolismABSTRACT
Ganoderma lucidum (Leyss.Fr.) Karst is one of the well-known medicinal macrofungi all over the world, and mounting researches have focused on the polysaccharides derived from the spores of G. lucidum. In the present study, BALB/c mice (n = 8-10) were administered with crude polysaccharides of G. lucidum spores (CPGS) and the refined polysaccharides of G. lucidum spores (RPGS) for 30 days to investigate their effect on the adaptive immune system. Results showed that CPGS and RPGS displayed diverse effects on the lymphocyte activity in the spleen. The splenocyte proliferation activity upon mitogen was suppressed by CPGS and RPGS, while the NK cell's tumor-killing ability was promoted by CPGS. Both CPGS and RPGS could increase the proportion of naïve T cells in thymus, but only RPGS significantly uplifted the percentage of T cells, as well as the T cell subsets, in peripheral blood, and promoted the activation by upregulating the expression of costimulatory factor CD28. Moreover, 16S sequencing results showed that the effects of CPGS and RPGS were closely related to the regulation of gut microbiota. ß-diversity of the microbiome was evidently changed by CPGS and RPGS. The phytoestrogen/polysaccharide-metabolizing bacteria (Adlercreutzia, Parabacteroides, and Prevotella), and an unclassified Desulfovibrionaceae, were remarkably enriched by CPGS or RPGS, and functions involving carbohydrate metabolism, membrane transport, and lipid metabolism were regulated. Moreover, the enrichments of Adlercreutzia, Prevotella, and Desulfovibrionaceae were positively related to the immune regulation by CPGS and RPGS, while that of Parabacteroides displayed a negative correlation. These findings suggested a promising effect of the polysaccharide from sporoderm-broken spore of G. lucidum in immune regulation to promote health control.
ABSTRACT
PLGA (poly(lactic-co-glycolic acid))-based nanoparticles (NPs) are promising drug carrier systems because of their excellent biocompatibility and ability for sustained drug release. However, it is not well understood how the kinetics of such drug delivery system perform in the retinal blood circulation as imaged in vivo and in real time. To answer this question, PLGA NPs were loaded either with lipophilic carbocyanine perchlorate (DiI) or hydrophilic Rhodamine 123 (Rho123) and coated with poloxamer 188 (P188): PLGA-DiI/P188 and PLGA-Rho123/P188. All particles had narrow size distributions around 130 nm, spherical shape and negative potential. Subsequently, we performed in vivo real-time imaging of retinal blood vessels, combined with ex vivo microscopy to monitor the kinetics and to detect location of those two fluorescent markers. We found that DiI signals were long lasting, detectable >90 min in blood vessels after intravenous injection as visible by homogeneous labelling of the vessel wall as well as by spots in the lumen of blood vessels. In contrast, Rho123 signals mostly disappeared after 15 min post intravenous injection in such compartment. To explore how PLGA NP-loaded cargoes are released in the retina in vivo, we thereafter monitored the Cyanine5.5 amine (Cy5.5) covalently linked PLGA polymer (Cy5.5-PLGA) in parallel to DiI and Rho123. The Cy5.5 signal from PLGA polymer was detectable in the retina vessels >90 min for both, the Cy5.5-PLGA-DiI/P188 and Cy5.5-PLGA-Rho123/P188 groups. Microscopy of the ex vivo retina tissue revealed partial level of colocalization of PLGA with DiI but no colocalization between PLGA and Rho123 at 2 h post injection. This indicates that at least a fraction of the lipophilic DiI was preserved within NPs, whereas no hydrophilic Rho123 was associated with NPs at that time point. In conclusion, the properties of PLGA carrier-cargo system in the blood circulation of the retina might be strongly influenced by the combination of factors, including the individual properties of loaded compounds and blood milieu. Thus, it is unlikely that a single nanoparticle formulation will be identified that is universally effective for the delivery of different compounds.
Subject(s)
Carbocyanines/metabolism , Drug Carriers , Fluorescent Dyes/metabolism , Microscopy, Confocal , Microscopy, Fluorescence , Nanoparticles , Polylactic Acid-Polyglycolic Acid Copolymer/chemistry , Retinal Vessels/metabolism , Rhodamine 123/metabolism , Animals , Carbocyanines/chemistry , Drug Compounding , Fluorescent Dyes/chemistry , Kinetics , Male , Poloxamer/chemistry , Rats , Regional Blood Flow , Rhodamine 123/chemistryABSTRACT
Development of functional materials capable of exhibiting chirality tunable circularly polarized luminescence (CPL) is currently in high demand for potential technological applications. Herein we demonstrate the formation of both left- and right-handed fluorescent helical superstructures from each enantiomer of a chiral tetraphenylethylene derivative through judicious choice of the solution processing conditions. Interestingly, both the aggregation induced emission active enantiomers exhibit handedness inversion of their supramolecular helical assemblies just by varying the solution polarity without any change in their molecular chirality. The resulting helical supramolecular aggregates from each enantiomer are capable of emitting circularly polarized light, thus enabling both right- and left-handed CPL from a single chiral material. The left- and right-handed supramolecular helical aggregates in the dried films have been characterized using spectroscopy, scanning electron microscopy, and transmission electron microscopy techniques. These new chiral aggregation induced emission compounds could find applications in devices where CPL of opposite handedness is required from the same material and would facilitate our understanding of the formation of helical assemblies with switchable supramolecular chirality.
ABSTRACT
Superparamagnetic iron oxide nanoparticles (SPIO-NPs) have great potential to be used in different pharmaceutical applications, due to their unique and versatile physical and chemical properties. The aim of this study was to quantify in vitro cytotoxicity of dextran 70,000-coated SPIO-NPs labelled/unlabelled with rhodamine 123, in C6 glioma cells and primary hippocampal neural cells. In addition, we analyzed the in vitro and in vivo cellular uptake of labelled SPIO-NPs. The nanoparticles, with average size of 10â»50 nm and polydispersity index of 0.37, were synthesized using Massart's co-precipitation method. The concentration-dependent cytotoxicity was quantified by using tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Intracellular localization of SPIO-NPs was detected by confocal laser microscopy. In vivo confocal neuroimaging (ICON) was performed on male Wistar rats after intravitreal injection followed by ex vivo retina whole mount analysis. When used for in vitro testing concentrations in the range of diagnostic and therapeutic dosages, SPIO-NPs proved to be non-cytotoxic on C6 glioma cells for up to 24 h incubation time. The hippocampal cell culture also did not show impaired viability at low doses after 24 h incubation. Our results indicate that our dextran-coated SPIO-NPs have the potential for in vivo drug delivery applications.
Subject(s)
Ferric Compounds/chemistry , Magnetite Nanoparticles/chemistry , Animals , Cell Line, Tumor , Cell Survival/drug effects , Glioma , Magnetite Nanoparticles/toxicity , RatsABSTRACT
A delivery platform with highly selective permeability through the blood-brain barrier (BBB) is essential for brain disease treatment. In this research, we designed and prepared a novel target nanoplatform, that is, layered double hydroxide (LDH) nanoparticle conjugated with targeting peptide-ligand Angiopep-2 (Ang2) or rabies virus glycoprotein (RVG) via intermatrix bovine serum albumin for brain targeting. In vitro studies show that functionalization with the target ligand significantly increases the delivery efficiency of LDH nanoparticles to the brain endothelial (bEnd.3) cells and the transcytosis through the simulated BBB model, that is, bEnd.3 cell-constructed multilayer membrane. In vivo confocal neuroimaging of the rat's blood-retina area dynamically demonstrates that LDH nanoparticles modified with peptide ligands have shown a prolonged retention period within the retina vessel in comparison with the pristine LDH group. Moreover, Ang2-modified LDH nanoparticles are found to more specifically accumulate in the mouse brain than the control and RVG-modified LDH nanoparticles after 2 and 48 h intravenous injection. All these findings strongly suggest that Ang2-modified LDHs can serve as an effective targeting nanoplatform for brain disease treatment.
Subject(s)
Nanoparticles , Animals , Blood-Brain Barrier , Brain , Hydroxides , Ligands , Mice , RatsABSTRACT
Much effort has been devoted to the discovery of effective biomaterials for nerve regeneration. Here, we reported a novel application of gold nanoparticles (AuNPs) modified with 6-mercaptopurine (6MP) and a neuron-penetrating peptide (RDP) as a neurophic agent to promote proliferation and neurite growth of human neuroblastoma (SH-SY5Y) cells. When the cells were treated with 6MP-AuNPs-RDP conjugates, they showed higher metabolic activity than the control. Moreover, SH-SY5Y cells were transplanted onto the surface coated with 6MP-AuNPs-RDP to examine the effect of neurite development. It can be concluded that 6MP-AuNPs-RDP attached to the cell surface and then internalized into cells, leading to a significant increase of neurite growth. Even though 6MP-AuNPs-RDP-treated cells were recovered from frozen storage, the cells still maintained constant growth, indicating that the cells have excellent tolerance to 6MP-AuNPs-RDP. The results suggested that the 6MP-AuNPs-RDP had promising potential to be developed as a neurophic nanomaterial for neuronal growth.
ABSTRACT
OBJECTIVE: There is 25-35% mutation rate of p53 in cancerous neural cells and this rate reaches 70-76% in glioma cell line. Complement of wild-type p53 has become a potential strategy for protein therapy of cancerous neural cells. Here we investigated the feasibility of a novel RDP-p53 fusion protein for anti-proliferation of cancerous neural cell and the possible mechanism, which would provide an effective approach for targeted delivery of p53 protein to treat cancerous neural cells. METHODS: The RDP-p53 fusion proteins are expressed in Escherichia coli, and they are labeled with FITC and rhodamine B by chemical modification. The fluorescence-labeled proteins are added to human hepatocellular carcinoma cells (HepG-2) and human neuroblastoma cells (SH-SY5Y) in order to investigate the possibility of RDP enhancing the cell uptake efficiency into neural cells as a cell-permeable carrier. The inhibitory effect of RDP-p53 on SH-SY5Y and human glioma cells (U251) was evaluated by MTT assay. Moreover, the anti-proliferation mechanism of RDP-p53 was determined by Apoptosis and Necrosis Assay Kit and flow cytometric analysis. RESULTS: The results showed that RDP-p53 could enter SH-SY5Y cells with high efficiency and selectively inhibit the growth of cancerous neural cells, including SH-SY5Y and U251. Also, cell apoptosis pathway and cell-cycle arrest at the G2/M phase were associated with the inhibition mechanism of RDP-p53 according to the data of flow cytometric analysis. CONCLUSIONS: RDP-p53 could be a novel antitumor candidate for targeting treatment of cancerous neural cells.
Subject(s)
Glioma/metabolism , Neuroblastoma/drug therapy , Tumor Suppressor Protein p53/chemistry , Tumor Suppressor Protein p53/metabolism , Apoptosis , Cell Line, Tumor , Cell Proliferation , Cell Survival , Glioma/chemistry , Humans , Neuroblastoma/chemistry , Neuroblastoma/metabolism , PermeabilityABSTRACT
Highly-efficient delivery of macromolecules into cells for both imaging and therapy (theranostics) remains a challenge for the design of a delivery system. Here, we suggested a novel hybrid protein-lipid polymer nanocapsule as an effective and nontoxic drug delivery and imaging carrier. The biodegradable nanocapsules showed the typical double emulsion features, including fluorescently labeled bovine serum albumin shell, oil phase containing poly(lactic-co-glycolic acid) and linoleic acid, and inner aqueous phase. The nanocapsules were spherical in shape, with an average size of about 180 nm. Proteins packed into the inner aqueous phase of the nanocapsules could be delivered into cells with high efficiency, and the fluorescence of the fluorescently labeled bovine serum albumin could be used for tracing the protein migration and cellular location. Further studies suggested that the co-delivery of transcription factor p53 and lipophilic drug paclitaxel with the nanocapsules acted synergistically to induce Hela cell apoptosis, and the fluorescence of apoptotic cells was clearly observed under a fluorescence microscope. Such multifunctional delivery system would have great potential applications in drug delivery and theranostic fields.
Subject(s)
Drug Carriers/chemistry , Nanocapsules/chemistry , Animals , Cattle , Cytological Techniques , Fluorescent Dyes/chemistry , HeLa Cells , Humans , Serum Albumin, BovineABSTRACT
Despite the significant progress in molecular imaging technologies that has been made in recent years, the specific detection of neural cells still remains challenging. Here, we suggest the use of gold nanoclusters (AuNCs) modified with a brain-targeting peptide as a potential imaging candidate for detecting neural cells in vitro and in mice. AuNCs of less than 10 nm (dynamic light scattering analysis) were first prepared using the "green" synthetic approach, and then a targeting peptide, rabies virus glycoprotein derived peptide (RDP), was conjugated to the AuNCs for improving the efficiency and specificity of neural cell penetration. The conjugate's mechanism of cellular attachment and entry into neural cells was suggested to be receptor-mediated endocytosis through clathrin-coated pits. Also, noninvasive imaging analysis and animal studies indicated that the RDP-modified nanoclusters could concentrate in the brain and locate in neural cells. This study suggests the feasibility of using targeting peptide-modified nanoclusters for noninvasive imaging brain cells in vivo.