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The main purpose of this study was the preparation, optimization, characterization, and in vitro immunological activity evaluation of the polysaccharides of Atractylodes macrocephala Koidz-loaded nanostructured lipid carriers (PAMK-NLC). Both the blank NLC and PAMK-NLC were prepared using melt-emulsification and ultra-sonication techniques. The optimization of preparation conditions were a stearic acid to caprylic/capric triglyceride mass ratio of 2:1, a poloxamer 188 to soybean lecithin mass ratio of 2:1, and an ultrasound time of 12 min; these parameters were identified using response surface methodology (RSM). Under these conditions, the encapsulation efficiency was around 76.85% and the polydispersity index was around 0.281, indicating a good dimensional distribution. Furthermore, the potential of PAMK-NLC to enhance immunological activity in vitro was investigated in murine splenic lymphocytes. The results showed that PAMK-NLC could significantly stimulate lymphocyte proliferation and increase the ratio of CD4+ to CD8+ T cells. In conclusion, these findings indicated that PAMK-NLC possessed an efficient immunological activity, and that NLC is suitable for the delivery and protection of PAMK in clinical use.

This paper develops a cooperative emergency inventory model to deal with supply disruptions. To achieve a cooperative state between the two sides in supply chain, the game theory is used and three coordination mechanisms are proposed. With the emergency inventory method which has been established, many tests of numerical experiments are made. The aim is to find the optimal mechanism so that the conditions of three mechanisms need to meet are analyzed. Results show that the total profits of the whole supply chain are unchanged in return mechanism. In cost-sharing mechanism, the manufacturer’s profits are smaller than the profits before coordination, and the supplier’s profits are greater than the former, but total profits of the whole supply chain become larger. The proposed model with the introduction of a second manufacturer mechanism performs best, which can make up the expenses by using the new joined manufacturers. It also can help the manufacturers respond to the supply disruptions effectively and that will not harm the profits of both sides.

In this study, angelica sinensis polysaccharide (ASP) was encapsulated into the Poly (lactic-co-glycolicacid) (PLGA) to constitute the ASP-PLGA. The aim of the study is to obtain optimal encapsulation efficiency of ASP-PLGA by optimizing the preparation conditions and investigate the immunological enhancement activity of ASP-PLGA. To obtain the optimal processing parameters, the parameters, such as the ratio of organic phase (o) to internal water phase (w1), the ratio of external water phase (w2) to organic phase (o) and the concentration of Pluronic F68 (F68) (w/v) were examined through response surface methodology (RSM). The optimum preparation conditions of ASP-PLGA for maximizing encapsulation efficiency (EE) was as follows:ratio of organic phase (o) to internal water phase (w1) at 10:1, ratio of external water phase (w2) to organic phase (o) at 11:1, the concentration of F68 at 0.8% and the experimental EE 67.89±0.48% was achieved. In addition, the results showed that ASP-PLGA could significantly promote the lymphocytes proliferation and increase the ratio of CD4+ to CD8+ T cells compared with ASP and blank PLGA. This study provided strong evidence that the immunological enhancement of ASP was significantly improved through establishing a drug delivery system with PLGA.

Port city is an important part of marine economic activities, whose development contributes to the connection of economic and environment between home and abroad. Sustainability of port cities not only can enhance the competitiveness of port cities, but also affects the city development process and characteristics. Therefore, it is pivotal to evaluate sustainability of port cities in advance. However, existing research on sustainable port cities is quite limited. And most of them are limited to the qualitative perspective, with little quantitative analysis. Therefore, this paper adopts a novel hybrid model THPD to evaluate sustainable port cities, expanding sustainability theories by taking port cities as research object. 21 port cities in China are selected for quantitative study. The results display that the sustainable level of port cities in China is at a medium level. The weaknesses and critical factors are varied, which points out the improvement direction for sustainable port cities.

Japan's decision to discharge nuclear wastewater into the sea on April 13, 2021 has aroused worldwide discussions recently. This paper establishes a tripartite evolutionary game model to explore the evolutionary stable strategies (ESSs) of Japan, other countries, and international environmental protection organization (IEPO). The results show: i) The choice of Japan is closely related to the treatment cost of nuclear wastewater. When Japan chooses the non-discharge strategy, other countries will always choose the acceptance strategy, while IEPO will trade off between its credibility and international assistance. When Japan chooses the discharge strategy, the choice of other countries depends on the difference of litigation compensation and cost, while that of IEPO is determined by the comparison of the litigation compensation and credibility with the litigation cost. ii) The treatment cost of nuclear wastewater, negative externalities of marine environment, litigation compensation, international assistance from IEPO, and the proximity coefficient between Japan and other countries are critical factors influencing both evolutionary results and trajectories, while the international assistance from other countries and initial willingness only affect evolutionary trajectories. This paper also puts forward policy implications for the three stakeholders to promote Japan's non-discharge strategy so as to protect the marine environment and human health from the unprecedented damage.

Shipping has become an essential mode of cargo transportation in the world and the number of global ships is also on the rise, along with the increasing marine environment pollution caused by ship waste. Various countries and regions have attached increasing importance to the pollution caused by ship waste in recent years. How to address ship waste management from international shipping has been brought to the official agenda of the international community. This article starts with the ship waste management framework in the international maritime sector and compares and analyzes the regulatory challenges of ship waste management faced by the International Maritime Organization (IMO) and typical countries and regions such as China, the United States, and the European Union, by analyzing the issues in their international maritime ship waste management regulations, including inconsistent and unclear standards, inadequate law enforcement efforts, and blind spots in high seas supervision. Targeting these challenges, this article proposes measures and directions for future supervision and stresses the necessity to unify ship waste discharge standards, improve port facilities, clarify standards, strengthen supervision and law enforcement, and establish a global monitoring system. This article lists the challenges of international maritime ship waste management and proposes practical countermeasures, which will help the international community better supervise ship waste discharge and reduce marine environment pollution.

To explore the regularity of ship density and ensure the safety of maritime traffic, a ship density model based on ship scale conversion and grid is established by considering the impact of ship size on traffic safety. A non-parametric kernel density estimation (KDE) method is used to obtain the probability density function of the ship density by selecting the appropriate kernel function and determining the optimal bandwidth. To process and store Automatic Identification System (AIS) data efficiently, a grid-based storage method of AIS data is proposed in this paper, and the relationship between grid width and sampling interval is established. The experiments result proves that the grid-based AIS data storage method saves 64% of storage space compared with the original AIS data. The ship density model based on ship scale conversion can reflect the impact of ship size on the maritime traffic. The distribution regularity of ship density can be obtained through the probability density curve of ship density and statistical characteristics. The research results show that the proposed method can provide theoretical support for the safety supervision of the maritime authority. It can also be recommended for the evaluation of VTS centers in vessel traffic management.

Nanoparticle delivery of functional molecules and vaccine is a promising method for enhancing the immune response. The objective of this study was to design chitosan (CS)-modified ginseng stem-leaf saponins (GSLS)-encapsulated cubosomes (Cub-GSLSCS) as a vaccine delivery system and explore its immunologic activity and adjuvanticity. In this study, CS-modified GSLS-encapsulated cubosomes (Cub-GSLSCS) were prepared. The storage stability of GSLS and that of ovalbumin (OVA) were measured. Additionally, the immunopotentiation of Cub-GSLSCS were assessed on potentiating macrophage in vitro, and the adjuvant activity was evaluated through immune response triggered by OVA model antigen. The encapsulation efficiency of optimized Cub-GSLSCS was about 65 % with Im3m nanostructure. The Cub-GSLSCS showed excellent stability and sustained release for up to 28 days. In vitro, Cub-GSLSCS nanoparticles improved cellular uptake, stimulated cytokines secretion of IL-6, IL-12, TNF-α, and generated more inducible nitric oxide synthase (iNOS) to produce higher levels of nitric oxide (NO) compared with other groups. Furthermore, the immunoadjuvant effects of OVA encapsulated Cub-GSLSCS nanoparticles (Cub-GSLSCS-OVA) were observed through immunized mice. Results showed that the ratio of CD4+/CD8 + T lymphocytes was increased in Cub-GSLSCS-OVA group. In addition, Cub-GSLSCS-OVA nanoparticles induced dramatically high OVA-specific IgG, IgG1, and IgG2a levels and stimulated the secretion of cytokines. Cub-GSLSCS may be a potential vaccine delivery system and induce a long-term sustained immunogenicity.

As the main component of international transportation, the shipping industry cannot be underestimated in pollutant emissions. To control the emission of pollutants, IMO and the major countries have designed ECA to limit ships’ emission in the offshore area. The implementation of ECA has a direct impact on the ship’s cost. Therefore, this paper studied how the ship’s behavior changes under ECA policy. Furthermore, we simulated the ship’s evasion trajectory under different ECA policies using AIS data. Meantime, the change of ship cost and emission reduction under different policies is analyzed. We found that the distorted emission reduction is increasing first and decreasing with the ECA boundary moving far away from the coastline. The saved cost by evasion behavior has the same variation trend. Furthermore, the emission reduction efficiency related to the increase in ship’s operating cost has a peak value with the distance of ECA boundary moving. This study could help the government to evaluate and design ECA policy.

Realizing the coordinated development of regional emission reduction and low-carbon transportation has become a hot issue concerned by governments worldwide. Using data from 30 provinces of China’s transport sector, this paper investigates and evaluates the regional differences of carbon emission intensity (CEI). Additionally, we employ the extended STIRPAT (stochastic impacts by regression on population, affluence and technology) and GTWR (geographically and temporally weighted regression) model to reveal the influence of driving factors on CEI from spatial-temporal perspectives. The results indicate the following: (1) CEI tends to decrease and presents a pattern of low in the southeast and high in the northwest in space. (2) The eastern region has the greatest degree of regional difference, and the central region has the smallest. The major reason for the widening of the overall differences in China is the within-group differences in the three geographical regions. (3) CEI in China’s transport sector has regional characteristics, and the effect of urbanization, energy structure, population scale and industrial structure on CEI vary among the regions. Energy intensity occupies a pivotal position among all the driving factors and poses a positive impact on the CEI of transport sector.

The success of subunit vaccines has been hampered by the problems of weak or short-term immunity and the lack of availability of nontoxic, potent adjuvants. It would be desirable to develop safe and efficient adjuvants with the aim of improving the cellular immune response against the target antigen. In this study, the targeting and sustained release of simple nanoliposomes containing Lycium barbarum polysaccharides (LBP) as an efficacious immune adjuvant to improve immune responses were explored. LBP liposome (LBPL) with high entrapment efficiency (86%) were obtained using a reverse-phase evaporation method and then used to encapsulate the model antigen, ovalbumin (OVA). We demonstrated that the as-synthesized liposome loaded with OVA and LBP (LBPL-OVA) was stable for 45 days and determined the encapsulation stability of OVA at 4°C and 37°C and the release profile of OVA from LBPL-OVA was investigated in pH 7.4 and pH 5.0. Further in vivo investigation showed that the antigen-specific humoral response was correlated with antigen delivery to the draining lymph nodes. The LBPL-OVA were also associated with high levels of uptake by key dendritic cells in the draining lymph nodes and they efficiently stimulated CD4+ and CD8+ T cell proliferation in vivo, further promoting antibody production. These features together elicited a significant humoral and celluar immune response, which was superior to that produced by free antigen alone.

Link prediction is a fundamental task in social networks, with the goal of estimating the likelihood of a link between each node pair. It can be applied in many situations, such as friend discovery on social media platforms or co-author recommendations in collaboration networks. Compared to the numerous traditional methods, this paper introduces utility analysis to the link prediction method by considering that individual preferences are the main reason behind the decision to form links, and meanwhile it also focuses on the meeting process that is a latent variable during the process of forming links. Accordingly, the link prediction problem is formulated as a machine learning process with latent variables; therefore, an Expectation–Maximization (EM, for short) algorithm is adopted and further developed to cope with the estimation problem. The performance of the present method is tested both on synthetic networks and on real-world datasets from social media networks and collaboration networks. All of the computational results illustrate that the proposed method yields more satisfying link prediction results than the selected benchmarks, and in particular, logistic regression, as a special case of the proposed method, provides the lower boundary of the likelihood function.

Background: Duck viral hepatitis (DVH), caused by duck hepatitis A virus (DHAV), is a fatal contagious infectious disease which spreads rapidly with high morbidity and high mortality, and there is no effective clinical drug against DVH.Materials and Methods: Raw Rehmannia Radix Polysaccharide (RRRP), Lycii Fructus polysaccharides and Astragalus Radix polysaccharides were experimented in vitro and in vivo. Mortality rate, livers change, liver lesion scoring, peroxidative injury evaluation indexes in vitro and in vivo, and hepatic injury evaluation indexes of optimal one were detected and observed in this experiment.Results: RRRP could reduce mortality with the protection rate about 20.0% compared with that of the viral control (VC) group, finding that RRRP was the most effective against DHAV. The average liver scoring of the VC, blank control (BC), RRRP groups were 3.5, 0, 2.1. Significant difference (P

Chinese angelica polysaccharides (CAP) and selenizing CAP (sCAP) were prepared and identified through FTIR and SEM observation. Their antioxidant activities in vitro and hepatoprotective effects in vivo were compared by free radical-scavenging tests or with CCl4-induced hepatic injury model mice. The results showed that for DPPH radical, superoxide anion and hydroxyl radical, the scavenging capabilities of sCAP were significantly stronger than those of CAP. In hepatic injury model mice, sCAP could significantly reduce ALT, AST and ALP contents and raised TP content in serum, significantly reduce MDA and ROS contents and raised SOD and T-AOC activities in liver homogenate in comparison with CAP; obviously relieve the pathological changes of liver and significantly inhibit the expressions of p-ERK, p-JNK and p-p38 protein as compared with those in model control group. These results indicate that selenylation modification can enhance the antioxidant and hepatoprotective actions of Chinese angelica polysaccharide. A action mechanism of sCAP is suppressing the protein expression of MAPK signaling pathway.

Context: The flavonoid prescription baicalin-linarin-icariin-notoginsenoside R1 (BLIN) has a curative effect on duck virus hepatitis (DVH) caused by duck hepatitis A virus type 1 (DHAV-1). However, the mechanism of this curative effect is not understood. Objective: This study investigates the mechanism of the curative effect of BLIN on DVH caused by DHAV-1. We analyzed the anti-DHAV-1 reproduction mechanism and immuno-regulatory effect of BLIN. Materials and methods: The anti-DHAV-1 reproduction effects of BLIN at 20, 10, 5 and 2.5 μg/mL in vitro, as well as the influence of BLIN at 20 μg/mL on DHAV-1 adsorption, replication and release were tested using the qRT-PCR method. The promotion abilities of BLIN at 20, 10, 5 and 2.5 μg/mL on T- and B-lymphocyte proliferation were investigated by the MTT method. IL-2 and IFN-γ levels and total anti-DHAV-1 antibody secretion after treatment with DHAV-1 for 4, 8 and 54 h were determined by ELISA. Results: BLIN showed a dose-dependent DHAV-1 reproduction inhibitory effect. The inhibitory effect was highest at 20 μg/mL, where DHAV-1 adsorption and release were significantly lower. Meanwhile, BLIN at 5 μg/mL significantly increased T and B lymphocyte proliferation. BLIN stimulated total anti-DHAV-1 antibody secretion in ducklings at the dosage of 4 mg per duckling, but did not stimulate IL-2 and IFN-γ secretion significantly. Conclusions: BLIN inhibits DHAV-1 reproduction by suppressing its adsorption and release. Additionally, BLIN promoted the duckling antiviral response.

Nanomedicine, the medical application of nanotechnology, promises a seemingly limitless range of applications from drug delivery to adjuvants and therapeutics. Our current research is focused on natural polymer-based liposome adjuvants. With the aim of inducing protective and long-lasting immunity, the immunological adjuvant activity of Rehmannia glutinosa polysaccharide liposome (RGPL) was investigated. In vivo, the splenic lymphocyte proliferation ratios and ovalbumin-specific immunoglobulin G titers of ovalbumin-RGPL-vaccinated mice were significantly upregulated. In draining lymph nodes, the expression of MHC II+CD11c+ and CD86+CD11c+ was increased by RGPL; in addition, the percentages of central memory cells (TCM) and effector memory cells (TEM) were also elevated. RGPL could effectively provide adequate antigen exposure in lymph nodes. In vitro, RGPL could promote dendritic cell maturation and enhance dendritic cell functions, such as the mixed lymphocyte reaction and antigen presentation. Overall, the results demonstrated that RGPL has the potential to act as an effective controlled release vaccine adjuvant.

Nanoparticle-based vaccine delivery systems have been extensively used to promote and induce immune responses to protein antigens. The properties of the nanoparticles, such as size, surface charge, and antigen loading mode, have been proved to significantly influence the adjuvant effect and immunoreactivity of nanoparticle-based vaccine delivery systems. The purpose of the study was to investigate how the surface charge and antigen loading mode of nanoparticles impact the immune responses. In this study, three ovalbumin (OVA)-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles with different surface charges and antigen loading modes were developed. The three nanoparticles were designed as antigen encapsulated with negatively charged (Angelica sinensis polysaccharide (ASP)-PLGA/OVA), antigen encapsulated with polyethylenimine (PEI)-coated (ASP-PLGA/OVA-PEI), and antigen adsorbed on PEI-coated (ASP-PLGA-PEI-OVA) nanoparticles. The Angelica sinensis polysaccharide (ASP) was used as the immunopotentiator and encapsulated into three nanoparticles. The results demonstrated that both PEI-coated (positively charged) nanoparticles promoted the antigen escape from the endosome, which led to the cytoplasmic antigen delivery to generate cross presentation, compared to negatively charged nanoparticles. In addition, PEI-coated nanoparticles activated the DCs in lymph nodes 5 days after the primary vaccination. In vivo experiments demonstrated that both antigen-encapsulated nanoparticles induced more potent and long-term antigen-specific antibody responses, compared to that of antigen-adsorbed nanoparticles. Thus, the PEI-coated and antigen-encapsulated nanoparticles (ASP-PLGA/OVA-PEI) as a vaccine adjuvant delivery system have the potential to induce strong and long-term humoral and cellular immune responses.

The post-disaster emergency medical rescue (EMR) is critical for people's lives. This paper presents a stochastic Petri net (SPN) model based on the process of the rescue structure and a Markov chain model (MC), which is applied to the optimization of the EMR process, with the aim of identifying the key activities of EMR. An isomorphic MC model is developed for measuring and evaluating the time performance of the EMR process during earthquakes with the data of the 2008 Wenchuan earthquake. This paper provides a mathematical approach to simulate the process and to evaluate the efficiency of EMR. Simultaneously, the expressions of the steady state probabilities of this system under various states are obtained based on the MC, and the variations of the probabilities are analyzed by changing the firing rates for every transition. Based on the concrete data of the event, the authors find the most time consuming and critical activities for EMR decisions. The model results show that the key activities can improve the efficiency of medical rescue, providing decision-makers with rescue strategies during the large scale earthquake.

In previous researches, the results showed that Alhagi honey polysaccharide-loaded poly(D,L-lactic-co-glycolic) acid nanoparticles (AHPP) as immune adjuvant enhanced Th1 immune responses. In order to further enhance the immune adjuvant activity and phagocytosis of the nanoparticles, three kinds of Alhagi honey polysaccharide-loaded cationic polymer modified PLGA nanoparticles were prepared to investigate the effects on macrophages in vitro. After treatment with the nanoparticles, the effects of phagocytosis, co-stimulatory molecules expression, nitric oxide (NO), inducible nitric oxide synthase (iNOS), and cytokines secretion were evaluated. The results showed that the surface structure of cationic polymer modified AHPP nanoparticles were not obviously changed, and the stability was greatly improved. Cationic polymer modified AHPP nanoparticles significantly stimulated phagocytic activity, MHCII+, CD86+, and CD80+ expression of macrophages. In addition, the levels of NO, iNOS, TNF-α, IFN-γ, IL-1β and IL-12 were enhanced in the peritoneal macrophages by stimulation with cationic polymer modified AHPP nanoparticles. Among them, polyethyleneimine modified PLGA nanoparticles (PEI-AHPP) showed the best effects on the expression of co-stimulatory molecules, and secretions of NO, iNOS, and cytokines. These results indicated that PEI-AHPP could enhance the activation of macrophages, and it could be potentially used as an AHP delivery system for the induction of strong immune responses.

Rabbit hemorrhagic disease (RHD) is an acute, high fatal contagious disease induced by rabbit hemorrhagic disease virus (RHDV) with acute severe hepatic injury and causes huge economic loss worldwide. In order to develop an effective and reliable drug to treat this disease in clinic, a prescription formulated with baicalin, linarin, icariin, and notoginsenoside R1 (BLIN) according to the theory of syndrome differentiation and treatment in traditional Chinese veterinary medicine was applied to investigate its curative effects against RHD in vivo. The preliminary study results showed that BLIN prescription exerted good curative effect on RHD therapy. To further validate the curative effect and to investigate the possible related curative mechanisms of this drug, the survival rates, the plasma biochemical indexes of hepatic function, the plasma evaluation indexes of oxidative injury, and the RHDV gene expression levels were detected and then the correlation among these indexes was also analyzed. These results showed that BLIN prescription could significantly increase the survival rate, reduce the hepatic injury severity, alleviate the oxidative injury, and decrease the RHDV gene expression level in rabbits infected with RHDV. All these results indicate that BLIN prescription possesses outstanding curative effect against RHD, and the curative mechanism may be related to its antioxidant and anti-RHDV activities. Therefore, this prescription can be expected to be exploited into a new candidate for RHD therapy in clinic.

Ports are an essential impetus for the development of cities, and cities provide important resources and infrastructure for ports. As a significant factor to enhance the competitiveness of port cities, the sustainable ability affects the development characteristics of cities. How to evaluate the sustainability has become the priority of port cities development. However, existing researches mainly focus on ports or cities separately, instead of combining the two. Therefore, we first establish a two-stage interaction model to associate port and city. Then we propose a new index system, covering the economic, environmental, and social indicators. Based on the evaluation framework, we adopt the directional distance function based on the slack-based measure model (SBM-DDF) and the coupling coordination degree (CCD) to measure the sustainability of port cities from the perspective of external influence and internal interaction. We select nine port cities in China as research objects. The results display their relative differences in sustainability. Hong Kong performs the best, while Xiamen performs the worst. According to the experience of Hong Kong, this paper puts forward corresponding improvement measures aiming at the disadvantage of port cities in order to achieve better sustainable development.

Five iriflophenone glycosides were obtained from the stems of Aquilaria sinensis by phytochemical methods. Their structures were determined by spectroscopic methods and comparison with literature data. As a novel compound, compound 1 was given a trivial name aquilarinoside B.

Hot topics, as a common phenomenon on social media platform, play a major role in public opinion. This paper aims to discuss the issues about the duration of hot topics: which factors influence the duration of a hot topic on a social media platform? To answer this question, Cox regression model of survival analysis was introduced to make empirical analysis. The survey data containing 60 hot topics from 2011 to 2013 was collected from a popular social media platform of China. Besides, model verification is implemented to validate the adopted model and the calibrated parameters. As a result, this paper finds that the number of participants and opinion leaders have a significant positive influence on an incident’s duration, and official response times have a significant negative impact; whereas, the number of attending media has no significant impact. Furthermore, the prediction accuracy in the validation is up to 85 %, which implies that the obtained results would be robust. The established model and the designed empirical analysis are demonstrated to be adaptable to analyze the duration of hot topics, and three factors are found to effect the duration of a hot topic based on our collected dataset. Our analytic method could also be adopted in numerous problems related to the duration issues in the field of information management.

Liposome-based vaccine delivery systems are known to enhance immune responses. Ganoderma lucidum polysaccharides (GLP) have been widely studied as immunomodulator and it could be as inducers of strong immune responses. In the research, GLP and ovalbumin (OVA) were encapsulated into liposome as vaccine and inoculated to mice. The magnitude and kinetics of the humoral and cellular immune responses were investigated. The results showed that GLP-OVA-loaded liposomes (GLPL/OVA) could induce more powerful antigen-specific immune responses than each single-component formulation. Mice immunized with GLPL/OVA displayed higher antigen-specific IgG antibodies, better splenocytes proliferation, higher cytokine secretion by splenocytes and significant activation of CD3+CD4+ and CD3+CD8+ T cells. Thus the GLPL/OVA formulation produced a heightened humoral and cellular immune response, with an overall Th1 bias. Enhanced immune responses elicited by the GLPL/OVA formulation might be attributed to effective activation and mature of DC in draining lymph nodes. Overall, these findings indicate that GLPL have the potential to enhance immune responses as vaccine delivery systems.

Duck hepatitis A virus type 1 (DHAV-1) is an important agent of duck viral hepatitis. Until recently, the replication cycle of DHAV-1 is still unknown. Here duck embryonic hepatocytes infected with DHAV-1 were collected at different time points, and dynamic changes of the relative DHAV-1 gene expression during replication were detected by real-time PCR. And the morphology of hepatocytes infected with DHAV was evaluated by electron microscope. The result suggested that the adsorption of DHAV-1 saturated at 90 min post-infection, and the virus particles with size of about 50 nm including more than 20 nm of vacuum drying gold were observed on the infected cells surface. What's more, the replication lasted around 13 h after the early protein synthesis for about 5h, and the release of DHAV-1 was in steady state after 32 h. The replication cycle will enrich the data for DVH control and provide the foundation for future studies.

One acidic polysaccharide named EPS-1 was isolated from the aqueous extract of the leaves of Epimedium acuminatum Franch. It may be composed of 1,4-linked α-d-GalpA, 1,3,4-linked α-d-GalpA, 1,6-linked β-d-Galp and terminal α-l-Rhap residues in a molar ratio of 11.0:1.0:1.0:1.0 by chemical and spectroscopic analysis. EPS-1 possessed immune modulation effects on peripheral T lymphocyte and immature chBM-DCs such as promoting the proliferation and cytokine secretion of these cells and increasing the phagocytosis ability of immature chBM-DCs.

The aim of this paper is to understand how disruptions effect on closed-loop supply chain, and how to achieve the coordination of closed-loop supply chain under disruptions situation for the whole supply chain system. Design/methodology/approach- This paper analyzed the effect caused by the disruptions of the closed-loop supply chain from both supply and demand , and applied the established margin profits sharing contract to achieve coordination under the disruptions. Findings- From both supply side and demand side, it drew how the disruptions affect the closed-loop supply chain. When the disruptions have less effect on the forward and reverse cost of closed-loop supply chain, the impact of the closed-loop supply chain is controllable; when the disruptions have lager effect on the forward and reverse cost of closed-loop supply chain, the impact of the closed-loop supply chain is uncontrollable. Research limitations/implications- In this paper, the contract costs are ignored for zero. While in actual situation, to maintain the efficient implementation of this contract must spend considerable cost. Sometimes the cost may be too high to become hinder between manufacturers and retailers. This paper also does not make in-depth study on this issue, so it will be further study in the future. Originality/value- This paper also designed a set of differential profit sharing contract based on revenue sharing contract mechanism. The contract is easy to be managed, easy to be operated, making the closed-loop supply chain system can achieve coordination whether in steady state or under disruptions.

The activation of murine peritoneal macrophages by Ganoderma lucidum polysaccharides liposomes (GLPL) was investigated in vitro. After treatment with GLPL, the changes of the nitric oxide (NO) secretion and iNOS (inducible nitric oxide synthase) activity were evaluated. The results showed that NO production and iNOS activity of macrophages were enhanced compared to GLP and BL group. In addition, both the phagocytic activity and levels of cytokines IL-1β, TNF-α and IFN-γ were enhanced in the peritoneal macrophages of mice by stimulation of GLPL. The expression of the major histocompatibility complex class II molecule (MHC II) on the surface of peritoneal macrophages significantly increased. These indicated that GLPL could enhance the activation of peritoneal macrophages and their potential for use as a delivery system of GLP.

Adjuvants improve vaccine potency by enhancing immunogenicity and sustaining long-term immune responses. Lentinan (LNT), a β-1,3-glucohexaose with β-1,6-branches, is extracted from the mushroom Lentinus edodes and functions as an effective immunostimulatory drug. Previous studies have demonstrated the adjuvant activity of calcium carbonate (CaCO3) microspheres as well as their use as antigen delivery systems. In this study, we successfully loaded CaCO3 microspheres with LNT and evaluated their physicochemical characteristics prior to the adsorption of ovalbumin. Our experimental results demonstrated that LNT-CaCO3 significantly enhanced lymphocyte proliferation, and boosted the frequency of CD69 + B cells and the ratio of CD4+ to CD8 + T cells in spleen lymphocytes. Moreover, LNT-CaCO3 unexpectedly induced the secretion of IgG and Th-associated cytokines (IL-2, IL-4, IFN-γ, and TNF-α) in immunized mice. Therefore, LNT-CaCO3 microspheres induce robust cellular and humoral immune responses and have potential utility as vaccine delivery systems.

In the shipping industry, forwarders can purchase a “one-stop” service from an ocean shipping (OS) company or a dual “ocean + inland” service from an OS company and an individual inland shipping (IS) company. Additionally, the OS company can strategically decide whether or not to enter the downstream market for cargo-canvassing, which will form a co-opetition relationship with incumbent forwarders. We investigate a strategy matrix based on the following two questions: “What kind of shipping service should be purchased by the forwarder?” “Is it beneficial for OS company to enter the market?” We find that when the forwarder’s cargo-canvassing effort level is low, a dual-service purchasing strategy will benefit both the stakeholders, while market-entry is a self-interested behavior and hurts the forwarder. Enhancing the cargo-canvassing effort level may help the forwarder to overcome this dilemma. However, with an increasing level of effort, there is a mutual inhibition of dual-service purchasing and market-entry, which hurts both the stakeholders’ profits simultaneously. Interestingly, we show that a win-win situation exists when the forwarder adopts a dual-service purchasing strategy without market-entry. Furthermore, we verify that this win-win situation is Pareto-optimized.

We previously reported that Chrysanthemum indicum polysaccharide (CIPS) effectively inhibited the replication of duck hepatitis A virus (DHAV). However, the inhibition mechanisms are still unclear. Autophagy plays important role in virus genomic replication. Therefore, in present study, the effect of autophagy on DHAV genome replication as well as the influence of CIPS on autophagy were studied. qPCR, western blot, and ELISA methods were applied to observe the autophagy and analyze the inhibition mechanisms of CIPS on DHAV. Results showed that DHAV infection increased the expression level of LC3-II and interdicted the degradation of p62. Treating with rapamycin benefited DHAV gene expression level. What's more, DHAV infection and rapamycin treatment also promoted the expression of PI3KC3 and increased the concentration of PI3P. However, CIPS treatment significantly downregulated the expressions of LC3-II and PI3KC3 induced by DHAV and rapamycin, and consequently inhibited autophagosomes formation. As a result, DHAV replication was inhibited.

Astragalus polysaccharides (APS) have long been well known as immune boosters, but have not been fully exploited in clinical settings. Here, poly(lactic-co-glycolic acid) (PLGA) was used to form a nanocarrier for APS to enhance its bioavailability. The aim was to improve the immunoadjuvanticity of conventional APS-loaded PLGA-based nanoparticles (NPs), referred to as APSPs, and to optimize the synthesis parameters to maximize the encapsulation efficiency (EE). As slow drug release can cause insufficient immune responses, ammonium bicarbonate was used to produce pH-responsive APSPs. The optimum parameters for maximizing EE (mean maximum experimental EE: 65.23 ± 0.51%) were an oil phase (O)/internal aqueous phase (W1) ratio of 7:1, an external aqueous phase (W2)/preliminary emulsion (PE) ratio of 5:1, and a Pluronic F-68 concentration of 1.1%. Moreover, the pH-responsive APSPs had low cytotoxicity and significantly enhanced mice splenic lymphocyte proliferation. The increased T-cell CD4+/CD8+ ratio after pH-responsive APSP treatment of mice splenic lymphocytes compared with free APS, blank PLGA NP, and conventional APSP treatment demonstrated its excellent immunoadjuvanticity. This study provides abundant evidence that these novel PLGA-based pH-responsive NPs enhanced the immunoadjuvanticity of APS. Furthermore, pH-responsive APSPs synthesized using the optimum parameters exhibited long-term stability in normal storage conditions, suggesting suitability for clinical application.

This study aimed to optimize the preparation conditions for cetyltrimethylammonium bromide-modified Polygonatum sibiricum polysaccharide cubosomes (CTAB-modified PSP-Cubs) by response surface methodology (RSM). Glyceryl-monooleate (GMO) was used as the lipid base for the cubosomes. The optimal preparation conditions of CTAB-modified PSP-Cubs were as follows: the mass percentage of PSP to GMO (X1), Poloxamer 407 (F127) to GMO (X2) and water to GMO (X3) was 1.4%, 9% and 50%, respectively. The encapsulation efficiency (EE) of CTAB-modified PSP-Cubs under the experimental conditions was 62.4 ± 3.6%, which was close to our predicted value. The particle size, polydispersity index (PDI) and zeta potential of CTAB-modified PSP-Cubs were 427.7 ± 8.0 nm, 0.236 ± 0.024 and 19.2 ± 0.4 mV, respectively. Specific modifications were able to slow down in vitro release behaviors and reduce cytotoxicity to some extent. The effects of CTAB-modified PSP-Cubs on splenic lymphocytes were also investigated. When splenic lymphocytes were treated with CTAB-modified PSP-Cubs together with LPS or PHA, the results showed more favorable effects on cellular proliferation than blank cubosomes or free PSP at certain concentrations.

Dendritic cells (DCs) are professional antigen-presenting cells (APC) that play a central role in the initiation and regulation of immune responses. We have previously demonstrated that Lycium barbarum polysaccharides liposomes (LBPL) as immune adjuvant elicits strong antigen-specific Th1 immune responses. The purpose of this study was to investigate underlying mechanism of liposomes promoting effect of Lycium barbarum polysaccharides (LBP) on activating DCs. LBP were loaded with high entrapment efficiency (86%) into liposomes using reverse phase evaporation. LBPL activation of phenotypic and functional maturation of DCs was explored through mechanistic studies of the TLR4-MyD88-NF-κB signaling pathway and amount of proinflammatory cytokines released. We found that LBPL indeed activated immature DCs and induced DCs maturation characterized by up-regulation of co-stimulatory molecules (MHCII, CD80, CD86), production of cytokines (IL-12p40, TNF-α), and enhancement of antigen uptake. Additionally, we demonstrated that liposomes could promote LBP up-regulation of TLR4, MyD88, TRAF6, NF-κB gene and protein expression.

Nanoparticles (NPs)-based vaccine delivery systems are widely used for their ability to control the release of antigens and promote immune responses against cancer or infectious diseases. In this study, the immunopotentiator Angelica sinensis polysaccharide (ASP) and model protein antigen ovalbumin (OVA) were encapsulated into Poly(lactic-co-glycolic acid) (PLGA) to formulate the novel NPs-based vaccine delivery system (ASP-PLGA/OVA). These formulations were subcutaneously administered to mice, then the magnitude and kinetics of antibody and cellular immune responses were assessed. The ASP-PLGA/OVA NPs were pherical in shape with smooth surfaces, approximately 225.2 nm in average size, negatively charged (around −11.27 mV), and the encapsulation efficiency of OVA at around 66.28%, respectively. Furthermore, ASP-PLGA/OVA NPs could keep stable at 4 °C over 30 days and provide a sustained and controlled release of OVA from the NPs. The results demonstrated that mice immunized with ASP-PLGA/OVA NPs could significantly enhance lymphocyte proliferation and improve the ratio of CD4+ to CD8+ T cells, thereby ASP-PLGA/OVA NPs could induce a strong cellular immune response. Moreover, the ASP-PLGA/OVA NPs could induce vigorous and long-term IgG immune responses with a mixed Th1 and Th2 responses and up-regulate the levels of Th-associated cytokines. These results suggested that ASP-PLGA/OVA NPs, which stimulated strong and continuous antibody responses and induced cellular immune responses, could potentially serve as an efficient and safe vaccine delivery and adjuvant system against infections and diseases.

The immunoregulation and immunopotentiation of Polysaccharides of Atractylodes macrocephala Koidz (PAMK) have been widely demonstrated. Nanostructured lipid carriers (NLC) have high drug loading capacity for lipophilic and hydrophilic drugs, and have good biocompatibility and high bioavailability. In this study, the effect of PAKM-NLC on the surface molecule expression of bone marrow-derived dendritic cells (BMDCs) in vitro was investigated by flow cytometry, and the cytokines secreted by dendritic cell supernatants were detected by ELISA. The results showed that compared with other control groups, PAMK-NLC could significantly increase the expression of CD80 and CD86 and promote the secretion of IL-1β, IL-12, TNF-α and IFN-γ, indicating that PAMK-NLC have a more pronounced effect on the maturation and differentiation of BMDCs. In addition, effects of PAMK-NLC nanoparticles on OVA-immunized mice were explored. Compared with other control groups, PAMK-NLC-OVA can significantly promote the production of OVA-specific antibodies in serum, stimulate the secretion of cytokines, increase the proliferation rate of spleen lymphocytes after OVA re-stimulation, and induce stronger activation of CD3+CD4+ and CD3+CD8+ lymphocytes. As an adjuvant of OVA, PAMK-NLC has a better immunological enhancement effect than PAMK or blank NLC, and has good adjuvant activity.

Background Duck viral hepatitis (DVH) is an acute disease of young ducklings with no effective veterinary drugs for treatment. Gynostemma pentaphyllum is a well-known traditional Chinese medicine that plays an important role in the treatment of various diseases. Gypenoside (GP), one of the main ingredients of Gynostemma pentaphyllum, was reported with good hepatoprotective effects. However, its low solubility limits its application in the clinics. To improve its solubility and bioactivity, a phosphorylated derivative of gypenoside (pGP) was prepared by the sodium trimetaphosphate-sodium tripolyphosphate (STMP-STPP) method. An infrared spectroscopy method was applied to analyse the structures of GP and pGP. Then, a methyl thiazolyl tetrazolium (MTT) colorimetric assay was applied to study the hepatocyte protective efficacy of these two drugs against duck hepatitis A virus type 1 (DHAV-1) infection, and qPCR, TUNEL labelling and flow cytometry methods were used to study the relevant hepatocyte protective in vitro. Results The infrared spectroscopy detection results showed that the phosphorylation modification of GP was successful. The MTT colorimetric assay results showed that both GP and pGP possessed good hepatocyte protective efficacy in vitro, and pGP performed better than GP when the drug was added before or after virus inoculation. Furthermore, the qPCR results revealed that both drugs could effectively inhibit the adsorption (when adding GP and pGP pre-virus inoculation), replication and release of DHAV-1, and the viral inhibition rate of pGP was greater than that of GP. The subsequent TUNEL labelling and flow cytometry assays showed that both GP and pGP could significantly inhibit duck embryo hepatocyte apoptosis induced by DHAV-1, and the inhibition effect of pGP was much stronger than that of GP. Conclusions GP exerts good hepatocyte protective efficacy not only by inhibiting the proliferation of DHAV-1 but also by inhibiting duck embryonic hepatocyte apoptosis induced by DHAV-1, and phosphorylation modification significantly improves the antiviral and the anti-apoptotic effects of GP. Therefore, pGP has the potential to be developed into a novel drug against DHAV-1 infection.

Duck hepatitis A virus type 1 (DHAV-1) is a serious infectious virus of ducklings. Recent study showed baicalin (BA) and baicalin phospholipid complex (BAPC) possessed anti-DHAV-1 effect. However, the antiviral mechanism is not clear. Therefore, the aim of the present work is to study influences and mechanisms of BA and BAPC on DHAV-1. The effects of BA and BAPC on DHAV-1 replication were analyzed by CCK-8 and RT-qPCR methods. And the results showed BA inhibited the replication of DHAV-1, and BAPC was more effective. Then, the influences of BA and BAPC on DHAV-1 protein translation and RNA synthesis were detected by western blot and RT-qPCR. Both BA and BAPC inhibited the protein translation, and BAPC did better. Furthermore, BAPC also inhibited the RNA synthesis. Afterwards, DHAV-1 IRES activity, DHAV-1 3D protein stability, and cellular Hsp70 expression were studied to in-depth understand the inhibition effects of BA and BAPC on DHAV-1 replication. The results indicated BA and BAPC dropped the protein translation via suppressing DHAV-1 IRES activity. Additionally, BAPC dropped the RNA synthesis via reducing the 3D protein stability and inhibiting cellular Hsp70 expression.

Sisi Zheng,1,* Tao Qin,2,* Yu Lu,3 Yifan Huang,2 Li Luo,1 Zhenguang Liu,1 Ruonan Bo,1 Yuanliang Hu,1 Jiaguo Liu,1 Deyun Wang1,2 1Institute of Traditional Chinese Veterinary Medicine, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 2Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal, Fujian Agriculture and Forestry University, Fuzhou, 3National Veterinary Product Engineering Research Center, Jiangsu Academy of Agricultural Sciences, Nanjing, People’s Republic of China *These authors contributed equally to this work Background: Poly lactide (PLA) was proved in the last years to be good for use in sustained drug delivery and as carriers for vaccine antigens. In our previous research, pachyman (PHY)-encapsulated PLA (PHYP) nanospheres were synthesized and their function of controlling drug release was demonstrated.Purpose: In order to modify the fast drug-release rate of PHY when inoculated alone, the maturation of bone marrow dendritic cells (BMDCs) in vitro and their immunological enhancement in vivo were explored using PHYP nanospheres.Methods: The maturation and antigen uptake of BMDCs were evaluated, both alone and with formulated antigen PHYP nanospheres, ie, ovalbumin (OVA)-loaded PHYP nanospheres, as an antigen delivery system, to investigate antigen-specific humoral and cellular immune responses.Results: The results indicated that, when stimulated by PHYP, the BMDCs matured as a result of upregulated expression of co-stimulatory molecules; the mechanism was elucidated by tracing fluorescently labeled antigens in confocal laser scanning microscopy images and observing the uptake of nanospheres by transmission electron microscopy. It was further revealed that mice inoculated with OVA-PHYP had augmented antigen-specific IgG antibodies, increased cytokine secretion by splenocytes, increased splenocyte proliferation, and activation of cluster of differentiation (CD)4+ and CD8+ T cells in vivo. Elevated immune responses were produced by OVA-PHYP, possibly owing to the activation and maturation of dendritic cells (in draining lymph nodes).Conclusion: It was corroborated that PHY- and/or OVA-encapsulated PLA nanospheres elicited prominent antigen-presenting effects on BMDCs and heightened humoral and cellular immune responses compared with other formulations. Keywords: PHYP, bone marrow dendritic cell, antigen delivery system, immune response

Dataset descriptions. (DOCX 16 kb)

Further comparison for other datasets. Table S1. Average performance in KNN and SVM classifiers of DKBCGS and KBCGS (two classification). Table S2. Average performance in KNN and SVM classifiers of our method and KBCGS (multi-classification). Table S3. Performance of gene feature selection methods with KNN classifier (high) and SVM classifier (low) in two-class datasets. (DOCX 27 kb)

To study the transfer of risk of dual channel supply chain in the electronic commerce environment, with the price of the risk transmission media, based on the cooperative game theory, The transmission of risk of supply chain in decentralized decision and centralized decision is analyzed in both supplier and retailer. The risk transmission mechanism of dual channel supply chain is researched in the emergence of sudden event, and the model of sudden risk transmission is established. Studies shows that the decision will affect the profit and the risk of the producer and seller, and the influence of producers and sellers is different; In general, the profit has a strong risk transfer to the market capacity and price sensitivity coefficient, has a fairly risky delivery to the consumer preference.

Existing gene selection methods: a brief introduction. (DOCX 18 kb)

SVM and KNN classifiers. (DOCX 18 kb)

Thirty-one compounds, including fourteen triterpenoids (1–14), seven coumarins (15–21), four lignans (22–25), and six diterpenes (26–31) were isolated from the aerial part of Euphorbia hirta. The structures were elucidated by spectroscopic methods and by comparison of their reported spectroscopic data. These compounds have shown the chemical relationships between this plant and other species from the genus Euphorbia.

A novel hemiterpene glycoside, urceolatoside A (1), together with known two hemiterpene glycosides (2, 3), two triterpenoid saponins (4, 5), and two aromatic compounds (6, 7), was isolated from the bark of Ilex urceolatus, a new species belonging to the genus Ilex. Their structures were elucidated as (2E)-2-methylbut-2-en-4-ol-1-O-[(5″-O-(7″E)-caffeoyl)-β-D-apiofuranosyl]-(1→6)-β-D-glucopyranoside (urceolatoside A) (1), aohada-glucoside C (2), pubescenoside B (3), ilexsaponin B1 (4), ilexsaponin B2 (5), p-hydroxyphenethyl-O-β-D-glucopyranoside (6), and 4-hydroxybenzoic acid (7) by spectroscopic methods. Among them, compound 1 was a novel hemiterpene glycoside; compounds 6 and 7 were isolated from this plant for the first time.

Twenty-nine compounds, including five acetophenone derivatives (1–5), three phenanthroindolizidine alkaloids (12–14), seven pentacyclic triterpenoids (15–21) and five C21 steroidal sapogenins (22–26), were isolated from the root of Cynanchum paniculatum (Bunge) Kitag. Their structures were determined by spectroscopic methods and comparison with reported data. Moreover, the chemotaxonomic relationships were also discussed. As a result, acetophenone derivatives, pentacyclic triterpenoids and C21 pregnane sapogenins can be recognized as chemotaxonomic markers for Cynanchum genus, and C. paniculatum has close relationships with some species of genus Cynanchum.

This paper studied the anti-duck hepatitis A virus (DHAV) activities of Astragalus polysaccharide (APS) and its sulfate (sAPS) compared with those of Bush Sophora Root polysaccharide (BSRPS) and its sulfate (sBSRPS). The antiviral activities of APS and sAPS were measured by MTT and real-time PCR methods, in vitro. In vivo experiment, the mortality rate and the evaluation indexes of hepatic injury, peroxidative injury and immune level were measured. Just like the condition of BSRPS and sBSRPS, the anti-DHAV activities of sAPS were stronger than those of APS, both in vitro and in vivo. It indicated sulfated modification could enhance the antiviral ability of polysaccharide. But unlike the antiviral effects of BSPRS and sBSRPS in vivo, APS and sAPS did not reduce the mortality rates as their abilities of scavenging free radicals and alleviating the hepatic injuries were weaker than those of BSRPS and sBSRPS. And they even did not enhance the immune levels.