Design, synthesis, and evaluation of the novel ozagrel-paeonol codrug with antiplatelet aggregation activities as a potent anti-stroke therapeutic agent.

Introduction: Ischemic stroke is the second most common chronic disease worldwide and is associated with high morbidity and mortality. Thromboembolism and platelet aggregation are the most characteristic features of stroke. Other than aspirin, no standard, accepted, or effective treatment for acute ischemic stroke has been established. Consequently, it is essential to identify novel therapeutic compounds for this condition. Methods: In this study, novel ozagrel/paeonol-containing codrugs were synthesized and characterized using 1H-NMR, 13C-NMR, and mass spectroscopy. Their antiplatelet aggregation activity was evaluated, with compound PNC3 found to exhibit the best effect. Subsequently, studies were conducted to assess its neuroprotective effect, pharmacokinetic properties and model its binding mode to P2Y12 and TXA2, two proteins critical for platelet aggregation. Results: The results indicated that PNC3 has good bioavailability and exerts protective effects against oxygen-glucose deprivation injury in PC12 cells. Molecular docking analysis further demonstrated that the compound interacts with residues located in the active binding sites of the target proteins. Conclusion: The codrugs synthesized in this study display promising pharmacological activities and have the potential for development as an oral formulation.

Phenotypic Variation in Flower Color and Morphology in the Gerbera (Gerbera hybrida) F1 Hybrid Population and Their Association with EST-SSR Markers.

Gerbera (Gerbera hybrida) is a widely cultivated ornamental plant. However, its genetic improvement is limited by the lack of genetic analysis and molecular markers for traits. In this study, we analyzed the phenotypic and genotypic variation of 140 F1 progeny from two gerbera varieties with different flower types and colors. We evaluated the flower's morphology, color, and pigment content of the F1 population and performed cluster principal component analysis (PCA) and correlation analysis. The results showed that the main ornamental traits of the hybrid progeny varied greatly. The segregation ratios of single and double flowers and ligulate and split ray florets were both 1:1. The flower colors of the F1 progeny were mainly red and purple-red, similar to the male parent's color. Furthermore, we conducted a genetic analysis of the hybrid progeny using EST-SSR markers and performed association analysis with phenotypic traits. We identified 2, 2, 3, 1, and 2 loci to be associated with peduncle length (PL), ray floret length (RFL), and outer ray floret; the level of apex relative to the top of involucre (LAI); outer corolla lips (OCL); and the b* of ray floret color, respectively. Our results reveal the genetic patterns of important ornamental traits and provide a theoretical basis and practical tools for gerbera genetic breeding.

An UPLC - Q - Orbitrap method for pharmacokinetics and tissue distribution of four triterpenoids in rats after oral administration of Poria cocos ethanol extracts.

Poria cocos (Schw.) Wolf, is a fungus that is widely used as medicine and dietary supplement in China. But its action mechanism is still not very clear. In this paper, a rapid, specific and sensitive high performace liquid chromatography coupled with hybrid quadrupole - orbitrap mass sepctrometry (UPLC - Q - Orbitrap MS) method has been developed and validated to simultaneously determine of four triterpenoids including Dehydrotumulosic acid (DTA), Dehydropachymic acid (DPA), Pachymic acid (PA), Dehydrotrametenolic acid (DMA) from Poria cocos in rat plasma and tissues. The analyte was extracted from rat plasma and tissue homogenates by protein precipitation with acetonitrile using glibenclamide as the internal standard (IS). Chromatographic separation was carried out on ACQUITY UPLC BEH - C18 column (2.1 mm × 50 mm, 1.7 µm) with a mobile phase composed of acetonitrile - water (containing 1.0 mmol/L ammonium acetate) using gradient elution at a flow rate of 0.2 mL/min. Electrospray ionization (ESI -) under negative ion mode was used, and its quantization was performed with multiple reaction monitoring (MRM) mode. The method was fully validated and successfully applied to pharmacokinetics and tissue distribution study in rats after oral administration of ethanol extracts of Poria cocos. Compared with that of plasma exporsure, triterpenoids could be detected in various tissues with a relatively high degree of tissue distribution. After oral administration, the concentration orders in seven different tissues were ranked as DTA > PA > DPA > DMA in intestine and stomach, wheras DTA > DMA > PA > DPA in heart, liver, spleen, lung and kidney tissues, which is speculated that DPA, PA may be converted into DMA in vivo. In conclusion, this results may provide a material basis for study of the pharmacological actions of triterpenoids in Poria cocos.

The difference of origin and extraction method significantly affects the intrinsic quality of licorice: A new method for quality evaluation of homologous materials of medicine and food.

As a homologous material of both medicine and food, licorice is a famous traditional Chinese medicine. In the application process, different origins and different extraction methods have an impact on the intrinsic quality of licorice. In order to ensure the intrinsic quality of commercially available licorice products, and to explore the influence of origin and extraction methods on the quality of licorice, we put forward a simple and effective discriminatory method for "chemometrics analysis-based fingerprint establishment". First, fingerprints of licorice ethanol extraction (LEE) and licorice water extraction (LWE) were established. Then, similarity analysis (SA), hierarchical clustering analysis (HCA), principal component analysis (PCA) and other chemometrics methods were used to select qualitative and quantitative markers. Besides, the quantitative determination of 7 compounds of licorice with statistical significance was carried out, which provided accurate and informative data for quality evaluation. Finally, discriminant analysis was used to trace the origin of licorice.

Classification and Association Analysis of Gerbera (Gerbera hybrida) Flower Color Traits.

Floral color plays a crucial role in plant life such as plant-pollinator interactions and modifying the abiotic environment of reproductive structures. In the current study, 123 gerbera accessions were divided into six color groups (white, yellow, orange, pink, red, and purple), based on Royal Horticultural Society Color Chart calibration and colorimeter measurement. Partial least squares discriminant analysis showed that the white group was mainly affected by L* value, a* value, C value, and total anthocyanin contents, while the yellow group was positively correlated with L* value, b* value, and total anthocyanin contents. Similarly, the orange group was mainly affected by b* value and total carotenoid contents, whereas the pink group was positively correlated with L* and h values. Furthermore, the red group was affected by L* value, a* value, C value, and total anthocyanin contents, whilst the purple group was mainly distributed by L* value, a* value, b* value, and total anthocyanin contents. Based on 'Jin Xiang' transcriptome data, 14,106 expressed sequence tag (EST)-SSR markers were identified and 48 pairs of primers (19 newly developed primers) were screened. Population genetic structure, neighbor-joining clustering, and principal coordinate analysis showed that 123 gerbera accessions could be divided into two groups. EST-SSR-based association analysis showed that 1, 1, 2, 1, 1, 2, and 1 significant loci were related to L*, a*, b*, C, and h, total carotenoid, and total anthocyanin contents, respectively. These results provide an important reference for flower color classification and genetic improvement of gerbera.

SDN enabled flexible optical data center network with dynamic bandwidth allocation based on photonic integrated wavelength selective switch.

Optical switching techniques featuring the fast and large capacity have the potential to enable low latency and high throughput optical data center networks (DCN) to afford the rapid increasing of traffic-boosted applications. Flexibility of the DCN is of key importance to provide adaptive and dynamic bandwidth to handle the variable traffic patterns generated by the heterogeneous applications while optimizing the network resources. Aiming at providing the flexible bandwidth for optical DCNs, we propose and experimentally investigate a software-defined networking (SDN) enabled reconfigurable optical DCN architecture based on novel optical top of rack (OToR) switch exploiting photonic-integrated wavelength selective switch. Experimental results show that the optical bandwidth per link can be automatically reallocated under the management of the deployed SDN control plane according to the variable traffic patterns. With respect to the network with inflexible interconnections, the average packet loss of the reconfigurable DCN decreases 1 order of magnitude and the server-to-server latency performance improves of 42.2%. Scalability investigation illustrates limited (11.7%) performance degradation as the reconfigurable network scale from 2560 to 40960 servers. Both the numerical and experimental assessments validate the proposed DCN with reconfigurable bandwidth feature and lower latency variations with respect to the inflexible DCNs.

Enhanced immune responses in pigs by DNA vaccine coexpressing GP3 and GP5 of European type porcine reproductive and respiratory syndrome virus.

The European (EU) type of porcine reproductive and respiratory syndrome virus (PRRSV) has recently emerged in China. In this study, three recombinant DNA vaccines, pVAX1-EU-ORF3-ORF5 (coexpressing EU type PRRSV GP3 and GP5), pVAX1-EU-ORF3 and pVAX1-EU-ORF5, were constructed and evaluated for their abilities to induce humoral and cellular responses as well as to protect piglets against homologous virus challenge. All piglets were given booster vaccinations at 21 days after the initial inoculation and then challenged 14 days later. Pigs inoculated with pVAX1-EU-ORF3-ORF5 developed significantly higher (P<0.05) PRRSV-specific antibody responses, neutralizing antibodies and levels of IL-4 and IL-10 than those given pVAX1-EU-ORF3, pVAX1-EU-ORF5 or pVAX1. Moreover, pigs immunized with pVAX1-EU-ORF3-ORF5 had markedly increased levels of IFN-γ and IL-2 in serum and T-lymphocytes (CD3(+)CD4(+) and CD3(+)CD8(+) T cells) in peripheral blood. Thus, EU-type PRRSV GP3 and GP5 proteins demonstrated good immunogenicity and reactogenicity and could induce cellular immunity in pigs. Following challenge with the Lelystad virus (LV) strain, piglets inoculated with pVAX1-EU-ORF3-ORF5 showed viremia and virus load distributed in organ tissues that were significantly lower (P<0.05) than those in the pVAX1-EU-ORF3 group and control group, and slightly lower than those in the pVAX1-EU-ORF5 group (P>0.05). As GP3 could enhance humoral- and cell-mediated immune responses to GP5, the results of this study suggested that these two proteins delivered by a vaccine can synergistically induce immunity against PRRSV.

Construction and immunogenicity of a DNA vaccine coexpressing GP3 and GP5 of genotype-I porcine reproductive and respiratory syndrome virus.

BACKGROUND: The European (EU) genotype of porcine reproductive and respiratory syndrome virus (Genotype-I PRRSV) has recently emerged in China. The coexistence of Genotype-I and -II PRRSV strains could cause seriously affect PRRSV diagnosis and management. Current vaccines are not able to protect against PRRSV infection completely and have inherent drawbacks. Thus, genetically engineered vaccines, including DNA vaccine and live vector engineered vaccines, have been developed. This study aimed to determine the enhanced immune responses of mice inoculated with a DNA vaccine coexpressing GP3 and GP5 of a Genotype-I PRRSV. RESULTS: To evaluate the immunogenicity of GP3 and GP5 proteins from European-type PRRSV, three DNA vaccines, pVAX1-EU-ORF3-ORF5, pVAX1-EU-ORF3 and pVAX1-EU-ORF5, were constructed, which were based on a Genotype-I LV strain (GenBank ID: M96262). BALB/c mice were immunized with the DNA vaccines; delivered in the form of chitosan-DNA nanoparticles. To increase the efficiency of the vaccine, Quil A (Quillaja) was used as an adjuvant. GP3 and GP5-specific antibodies, neutralizing antibodies and cytokines (IL-2, IL-4, IL-10 and IFN gamma) from the immunized mice sera, and other immune parameters, were examined, including T-cell proliferation responses and subgroups of spleen T-lymphocytes. The results showed that ORF3 and ORF5 proteins of Genotype-I PRRSV induced GP3 and GP5-specific antibodies that could neutralize the virus. The levels of Cytokines IL-2, IL-4, IL-10, and IFN-γ of the experimental groups were significantly higher than those of control groups after booster vaccination (P < 0.05). The production of CD3+CD4+ and CD3+CD8+ T lymphocyte was also induced. T lymphocyte proliferation assays showed that the PRRSV LV strain virus could stimulate the proliferation of T lymphocytes in mice in the experimental group. CONCLUSIONS: Using Quil A as adjuvant, Genotype-I PRRSV GP3 and GP5 proteins produced good immunogenicity and reactivity. More importantly, better PRRSV-specific neutralizing antibody titers and cell-mediated immune responses were observed in mice immunized with the DNA vaccine co-expressing GP3 and GP5 proteins than in mice immunized with a DNA vaccine expressing either protein singly. The results of this study demonstrated that co-immunization with GP3 and GP5 produced a better immune response in mice.