Correlation of Folic Acid Metabolic Gene Polymorphisms, Homocysteine, Vitamin B12, and Red Blood Cell Folate with Adverse Pregnancy Outcomes.

Objective: This study aims to investigate the relationship between folic acid (FA) metabolic gene polymorphisms, homocysteine (Hcy), vitamin B12 (Vit B12), and red blood cell folate (RBCF) with adverse pregnancy. The findings of this study can help in the prevention and treatment of adverse pregnancy in the future. Methods: 118 pregnant women admitted to Qingdao Central Hospital between August 2020 and October 2022 were selected for retrospective analysis, including 62 cases of normal delivery (control group, CG) and 56 cases of adverse pregnancy (research group, RG). The single nucleotide polymorphisms of MTHFR C677T, MTHFR A1298C, and MTRR A66G gene loci were tested in both cohorts. Besides, differences in Hcy, Vit B12, and RBCF levels were observed, as well as Hcy, Vit B12, and RBCF alterations in different genotype carriers in the research group. Results: An elevated proportion of MTHFR 677TT-type gene and MTRR 66GG-type gene carriers and a lower proportion of MTRR 66GG-type gene carriers were found in the research group (χ2 = 4.458, 4.238, 4.206, P = .035, .040, .040). As indicated by the Logistic regression analysis, carriers of MTHFR 677TT and MTRR 66GG gene had an increased risk of adverse pregnancy outcomes (95%CI=2.881-5.942, 1.427-3.809, P < .001), while MTRR 66AG carriers had a decreased risk (95%CI=0.124-1.849, P < .001). Finally, Hcy levels of MTHFR 677TT and MTRR 66GG gene carriers increased, while Vit B12 and RBCF decreased; the opposite was true for MTRR 66AG gene carriers (P < .001). Conclusions: FA metabolic gene polymorphisms, Hcy, Vit B12, and RBCF are closely related to adverse pregnancy outcomes, which is of great significance for future clinical evaluation of adverse pregnancy.

Extraction, separation and kinetics of phenylethanosides from Plantago asiatica L. by an innovative extraction technology-deep eutectic solvent-based ultrasound-assisted extraction.

In this paper, the total phenylethanosides (TPS) were extracted efficiently by an innovative extraction technology--deep eutectic solvent-based ultrasound-assisted extraction (DES-UAE) from Plantago asiatica L. Ten diverse types of DESs were synthesized as alternative extraction solutions. The extraction efficiency of DES-3 (constituted by choline chloride and lactic acid) was much higher than those of other DESs. On the basis of single factor tests and Box-Behnken design (BBD), the optimum processing parameters of DES-UAE as follow: DES-3 with molar ratio of 1:3, extraction temperature 51 °C, solid/liquid 22.5 mg/ml, water content 30%, ultrasonic power 65 W, extraction time 23 min. The extraction efficiency of TPS from Plantago asiatica L. was 8.395 mg/ml, which was more superior than those of organic solvents (water, methanol, 50% methanol, ethanol, 50% ethanol). The extraction kinetics experiment results showed that water content had a significant influence upon the extraction efficiency of TPS. At the same time, AB-8 macroporous resin column was used to efficiently isolate TPS from DES extraction with a recovery rate of 88.5%.

Choline alleviated perinatal fluoride exposure-induced learning and memory impairment through α4ß2 nAChRs and α7 nAChRs in offspring mice.

Fluoride pollution is widely present in the living environment. As a critical period of brain development, the perinatal period is extremely vulnerable to fluoride. Studies have found that choline can protect the brain's memory and enhance the ability to focus. However, the effect of choline on perinatal fluoride-induced nerve damage remains unclear. Therefore, 32 Kunming newly conceived female mice and their offspring mice were randomly divided into control, NaF, LC + NaF, and HC + NaF groups, and the HE staining, Y-maze test, RT-PCR, western blotting, immunohistochemistry, etc. were used in this study. The results showed that fluoride decreased the brain organ coefficients and brain protein content (p < 0.05, p < 0.01), and caused histomorphological damage in the hippocampus and cortex, which suggested that fluoride affected the development of the brain and damaged the brain. Moreover, the results of the Y-maze test showed that fluoride increased the number of learning days, error reaction time, and total reaction time, and decreased the AchE activity in the brain (p < 0.05, p < 0.01), which indicated that fluoride reduced the learning and memory ability of the mice. Besides, the results showed that fluoride decreased the mRNA and protein expression levels of α4ß2 nAChRs and α7 nAChRs in the hippocampus and cortex (p < 0.05, p < 0.01). However, perinatal choline supplementation reversed the aforementioned fluoride-induced changes. In short, these results demonstrated that choline alleviated perinatal fluoride-induced learning and memory impairment, which will provide a rationale for the mitigation and prevention of fluoride-induced brain damage.

Optimization and Kinetics Modeling of Microwave-Assisted Subcritical n-Butane Extraction of Tigernut Oil.

In this study, tigernut oil was extracted from tigernut meal by subcritical n-butane extraction with the assistance of microwave pretreatment. Effects of microwave pulse duration, particle size of tigernut meal, and subcritical extraction variables (temperature, time, solid-liquid ratio, number of extraction cycles) on extraction efficiency were examined by single-factor experiments and Response Surface Methodology (RSM) modeling. The results indicate that microwaving (560 W, 6 min) significantly increased the subcritical extraction efficiency. The variation of extraction yield could be interpreted as a nonlinear function of extraction time, temperature and liquid-solid ratio. Changing the independent variables could affect the oil extraction efficiency. The subcritical extraction of tigernut oil with a liquid-solid ratio of 3.62 kg/(kg of tigernut meal) at a temperature of 52°C for 32 min after three extraction cycles produced the most oil, and a maximum yield (24.736%) of tigernut oil was achieved. The ratio of unsaturated to saturated fatty acids (4.68 UFA/SFA), low acid value (3.30 mg KOH/g oil), low peroxide value (0.28 meq.kg-1), and preponderance of oleic acid indicate a high-quality oil. To describe the extraction kinetics, a modified Brunner's mathematical model was used. The model fit the experimental data well over the entire operating range, and the explanation coefficient exceeds 96%. Our results can be used to develop an optimized method for subcritical fluid extraction of tigernut oil and can move industry further toward implementing microwave-assisted subcritical extraction in oil processing.

Early-Onset Diabetes Mellitus in Chromosome 8p11.2 Deletion Syndrome Combined With Becker Muscular Dystrophy - A Case Report.

Background: Chromosome 8p11.2 includes several key genes in development such as the FGFR1, ANK1, KAT6A, and SLC20A2 genes. Deletion of this fragment causes a contiguous gene syndrome. Currently, few cases of interstitial deletion of whole 8p11.2 have been reported. We report a rare case of 8p11.2 deletion syndrome with the unique phenotypes, presenting with early-onset diabetes. Case Description: A 20-year-old man with a 1-year history of diabetes mellitus was admitted to the Endocrinology Clinic. Physical examination revealed the dysmorphic facial features, and broad and foreshortened halluces. Laboratory examination indicated spherocytosis anemia, and hypogonadotropic hypogonadism. Bone mineral density analysis showed decreased bone density in the lumbar vertebrae. Brain CT showed calcification. Whole-exome sequencing revealed a 7.05-Mb deletion in 8p11 containing 43 OMIM genes, and a large in-frame deletion of exons 48-55 in the DMD gene. Metformin was given to the patient after which his blood glucose was well controlled. HCG was injected subcutaneously and was supplemented with calcium and vitamin D, which led to an improvement in the patient's quality of life. Conclusion: We report a rare case of 8p11.2 deletion syndrome with unique phenotypes, and early-onset diabetes. It is challenging for endocrinologists to simultaneously reconcile a combination of these diseases across multiple disciplines. We discussed the influencing factors of early-onset diabetes in this patient and speculated that it was caused by complex interactions of known and unknown genetic backgrounds and environmental factors.

Petroleum extract of Farfarae Flos alleviates nasal symptoms by regulating the Th1-Th2 cytokine balance in a mouse model of Allergic Rhinitis.

Farfarae Flos is a traditional Chinese medicine that has long been used to treat allergies. In this study, we aimed to investigate the effect of a petroleum extract of Farfarae Flos (PEFF) in a mouse model of allergic rhinitis (AR) and to explore the underlying molecular mechanisms of action. An animal model of AR was established by sensitization and challenge of BALB/c mice with ovalbumin (OVA). PEFF was administered intranasally and AR nasal symptoms were assessed on a semi-quantitative scale according to the frequencies of nose rubbing and sneezing and the degree of rhinorrhea. The mechanism of action of PEFF was evaluated by histological analysis of nasal mucosa architecture and inflammatory status; ELISA-based quantification of serum OVA-specific IgE, interferon-γ (IFN-γ), and interleukin-4 (IL-4) concentrations; and immunohistochemical and western blot analysis of T-bet and GATA3 protein expression in nasal mucosa and spleen tissues. The results showed intranasal administration of PEFF alleviated AR symptom scores and reduced both the infiltration of inflammatory cells and tissue damage in the nasal mucosa. PEFF significantly decreased serum concentrations of OVA-specific IgE (P<0.01) and IL-4 (P<0.05) and significantly increased IFN-γ (P<0.01). PEFF also upregulated the expression of T-bet protein (P<0.05) but downregulated GATA3 protein (P<0.05) in nasal mucosa and spleen tissues. In conclusion, PEFF effectively reduces AR nasal symptoms and serum IgE levels in a mouse model and may act by correcting the imbalance between Th1 and Th2 responses.

Polydatin inhibits ZEB1-invoked epithelial-mesenchymal transition in fructose-induced liver fibrosis.

High fructose intake is a risk factor for liver fibrosis. Polydatin is a main constituent of the rhizome of Polygonum cuspidatum, which has been used in traditional Chinese medicine to treat liver fibrosis. However, the underlying mechanisms of fructose-driven liver fibrosis as well as the actions of polydatin are not fully understood. In this study, fructose was found to promote zinc finger E-box binding homeobox 1 (ZEB1) nuclear translocation, decrease microRNA-203 (miR-203) expression, increase survivin, activate transforming growth factor ß1 (TGF-ß1)/Smad signalling, down-regulate E-cadherin, and up-regulate fibroblast specific protein 1 (FSP1), vimentin, N-cadherin and collagen I (COL1A1) in rat livers and BRL-3A cells, in parallel with fructose-induced liver fibrosis. Furthermore, ZEB1 nuclear translocation-mediated miR-203 low-expression was found to target survivin to activate TGF-ß1/Smad signalling, causing the EMT in fructose-exposed BRL-3A cells. Polydatin antagonized ZEB1 nuclear translocation to up-regulate miR-203, subsequently blocked survivin-activated TGF-ß1/Smad signalling, which were consistent with its protection against fructose-induced EMT and liver fibrosis. These results suggest that ZEB1 nuclear translocation may play an essential role in fructose-induced EMT in liver fibrosis by targeting survivin to activate TGF-ß1/Smad signalling. The suppression of ZEB1 nuclear translocation by polydatin may be a novel strategy for attenuating the EMT in liver fibrosis associated with high fructose diet.

Polygonum cuspidatum extract attenuates fructose-induced liver lipid accumulation through inhibiting Keap1 and activating Nrf2 antioxidant pathway.

BACKGROUND: Polygonum cuspidatum has been used in traditional Chinese medicine to treat liver disorders associated with oxidative stress, inflammation and lipid accumulation for centuries in patients. PURPOSE: The aim of this study was to examine whether P. cuspidatum extract (PCE) prevented against fructose-induced liver lipid accumulation via regulating Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. METHOD: PCE was administered orally to male Sprague-Dawley rats given 10% fructose drinking water for 6 weeks at 80 and 160 mg/kg once daily for 11 weeks. RESULTS: PCE significantly alleviated liver lipid accumulation in fructose-fed rats with metabolic syndrome. It also inhibited Keap1, activated Nrf2 antioxidant pathway, resulting in the suppression of oxidative stress, evidenced by reducing hydrogen peroxide (H2O2), malondialdehyde (MDA) and hydroxy radical (OH•) levels, and increasing glutathione (GSH)/oxidized glutathione (GSSG) ratio as well as superoxidase dismutase (SOD) and catalase (CAT) activity in the liver of fructose-fed rats. Additionally, PCE up-regulated peroxisome proliferator activated receptor-α (PPAR-α), and down-regulated sterol regulatory element binging protein 1 (SREBP-1), fatty acid synthetase (FAS) and stearoyl-CoA desaturase-1 (SCD-1) in this animal model, being consistent with its reduction of triglyceride (TG) levels. CONCLUSION: These results demonstrate that PCE reduces oxidative stress, and prevent lipid accumulation in the liver of fructose-fed rats possibly by targeting the Keap1/Nrf2 pathway. PCE may be a promising therapeutic strategy for fructose-associated liver lipid accumulation.

Magnesium isoglycyrrhizinate ameliorates fructose-induced podocyte apoptosis through downregulation of miR-193a to increase WT1.

High fructose intake is a risk of glomerular podocyte dysfunction. Podocyte apoptosis has emerged as a major cause of podocyte loss, exacerbating proteinuria. Magnesium isoglycyrrhizinate (MgIG) is usually used as a hepatoprotective agent in clinic. Liver and kidney injury often occurs in human diseases. Recent report shows that MgIG improves kidney function. In this study, we found that MgIG significantly alleviated kidney dysfunction, proteinuria and podocyte injury in fructose-fed rats. It also restored fructose-induced podocyte apoptosis in rat glomeruli and cultured differentiated podocytes. Of note, high-expression of miR-193a, downregulation of Wilms' tumor protein (WT1) and RelA, as well as upregulation of C-Maf inducing protein (C-mip) were observed in these animal and cell models. The data from the transfection of miR-193a mimic, miR-193a inhibitor, WT1 siRNA or LV5-WT1 in cultured differentiated podocytes showed that fructose increased miR-193a to down-regulate WT1, and subsequently activated C-mip to suppress RelA, causing podocyte apoptosis. These disturbances were significantly attenuated by MgIG. Taken together, these results provide the first evidence that MgIG restrains fructose-induced podocyte apoptosis at least partly through inhibiting miR-193a to upregulate WT1, supporting the application of MgIG with a novel mechanism-of-action against podocyte apoptosis associated with fructose-induced kidney dysfunction.

Purification and characterization of a glucose-tolerant ß-glucosidase from black plum seed and its structural changes in ionic liquids.

The objective of this study was to characterize a plant origin ß-glucosidase from black plum seeds and identify its conformational changes in twenty-six imidazolium- and amino acid-based ionic liquids (ILs). The results revealed that the purified 60 kDa enzyme was monomeric in nature, maximally active at 55 °C and pH 5.0, and nearly completely inhibited by Hg2+ and Ag+. Attractive peculiarities of the relative low kinetic and higher glucose inhibition constants (Km = 0.58 mM [pNPG]; Ki = 193.5 mM [glucose]) demonstrated its potential applications in food industry. Circular dichroism studies showed that the secondary structural changes of the enzyme depended not only on the anions, but also on the cations of the assayed ILs. Interestingly, no corresponding relations were observed between the changes in enzyme structure induced by ILs and its catalytic activities, suggesting that the influences of ILs on enzymatic processes don't rely simply on enzyme conformational changes.

Polydatin prevents fructose-induced liver inflammation and lipid deposition through increasing miR-200a to regulate Keap1/Nrf2 pathway.

Oxidative stress is a critical factor in nonalcoholic fatty liver disease pathogenesis. MicroRNA-200a (miR-200a) is reported to target Kelch-like ECH-associated protein 1 (Keap1), which regulates nuclear factor erythroid 2-related factor 2 (Nrf2) anti-oxidant pathway. Polydatin (3,4',5-trihydroxy-stilbene-3-ß-D-glucoside), a polyphenol found in the rhizome of Polygonum cuspidatum, have anti-oxidative, anti-inflammatory and anti-hyperlipidemic effects. However, whether miR-200a controls Keap1/Nrf2 pathway in fructose-induced liver inflammation and lipid deposition and the blockade of polydatin are still not clear. Here, we detected miR-200a down-regulation, Keap1 up-regulation, Nrf2 antioxidant pathway inactivation, ROS-driven thioredoxin-interacting protein (TXNIP) over-expression, NOD-like receptor (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome activation and dysregulation of peroxisome proliferator activated receptor-α (PPAR-α), carnitine palmitoyl transferase-1 (CPT-1), sterol regulatory element binging protein 1 (SREBP-1) and stearoyl-CoA desaturase-1 (SCD-1) in rat livers, BRL-3A and HepG2 cells under high fructose induction. Furthermore, the data from the treatment or transfection of miR-200a minic, Keap1 and TXNIP siRNA, Nrf2 activator and ROS inhibitor demonstrated that fructose-induced miR-200a low-expression increased Keap1 to block Nrf2 antioxidant pathway, and then enhanced ROS-driven TXNIP to activate NLRP3 inflammasome and disturb lipid metabolism-related proteins, causing inflammation and lipid deposition in BRL-3A cells. We also found that polydatin up-regulated miR-200a to inhibit Keap1 and activate Nrf2 antioxidant pathway, resulting in attenuation of these disturbances in these animal and cell models. These findings provide a novel pathological mechanism of fructose-induced redox status imbalance and suggest that the enhancement of miR-200a to control Keap1/Nrf2 pathway by polydatin is a therapeutic strategy for fructose-associated liver inflammation and lipid deposition.

An efficient method for decoloration of polysaccharides from the sprouts of Toona sinensis (A. Juss.) Roem by anion exchange macroporous resins.

An efficient decoloration method for polysaccharides from the sprouts of Toona sinensis (A. Juss.) Roem (PSTS) by anion exchange macroporous resins (AEMR) was investigated in the present paper. The results suggested that D941 resin offered better decoloration efficiency than other tested resins. Based on single-factor experiments, the optimal decoloration parameters of D941 resin were obtained as follows: temperature of 45°C, sample initial concentration of 30mg/ml, pH value of 8.5, static decoloration time of 90min, dynamic decoloration processing volume of 5.5BV with the flow rate of 2BV/h. Decoloration ratio, PSTS recovery ratio and selectivity coefficient were 91.94±1.23%, 90.05±2.35% and 10.92±0.63, respectively. Most of pigment impurities were successfully removed from PSTS solutions after treated by D941 resin, and there was no significant difference in carbohydrate concentration, characteristic groups and molecular weight. Compared with H2O2 oxidation and activated carbon adsorption, this developed method is superior.

Optimization on Preparation Conditions of Salidroside Liposome and Its Immunological Activity on PCV-2 in Mice.

The aim of this study was to optimize the preparation conditions of salidroside liposome with high encapsulation efficiency (EE) and to study the immunological enhancement activity of salidroside liposome as porcine circovirus type 2 virus (PCV-2) vaccine adjuvant. Response surface methodology (RSM) was selected to optimize the conditions for the preparation of salidroside liposome using Design-Expert V8.0.6 software. Three kinds of salidroside liposome adjuvants were prepared to study their adjuvant activity. BALB/c mice were immunized with PCV-2 encapsulated in different kinds of salidroside liposome adjuvants. The PCV-2-specific IgG in immunized mice serum was determined with ELISA. The results showed that when the concentration of ammonium sulfate was 0.26 mol·L(-1), ethanol volume 6.5 mL, temperature 43°C, ethanol injection rate 3 mL·min(-1), and salidroside liposome could be prepared with high encapsulation efficiency of 94.527%. Salidroside liposome as adjuvant could rapidly induce the production of PCV-2-specific IgG and salidroside liposome I adjuvant proved to provide the best effect among the three kinds of salidroside liposome adjuvants.

Adjuvanticity of compound astragalus polysaccharide and sulfated epimedium polysaccharide per os.

On the basis of previous researches, compound astragalus polysaccharide (APS) and sulfated epimedium polysaccharide (sEPS) oral liquid (AEO) was prepared. Three hundred and twenty 14-day-old chickens were randomly assigned into eight groups and vaccinated with ND vaccine except for blank control (BC) group, repeated vaccination at 28 days old. At the same time of each vaccination, the chickens in three experimental groups were taken orally with AEO, respectively, at three doses, in two component control groups with APS and sEPS, once a day for three successive days; in injection control group were injected with AEI once, and in vaccination control (VC) and BC groups were not administrated. On days 7, 14, 21, 28 and 35 after the first vaccination, peripheral lymphocyte proliferation, the serum antibody titer, IFN-γ and IL-2 concentrations and on day 35 immune organ index were measured. The results showed that AEO at high and medium doses could significantly promote lymphocyte proliferation and development of immune organ, enhance antibody titer and IFN-γ and IL-2 concentration, which was stronger than actions of AEI and two components. The results confirmed that AEO possessed reliable immunoenhancement and could be exploited into an oral immunopotentiator.

The immunological adjuvant activity of gypenosides liposome against Newcastle disease vaccine.

The adjuvant activity of gypenosides liposome (GPSL) encapsulated with liposome was investigated in vitro and in vivo. In vitro, different concentrations of GPSL were added into chicken's peripheral blood lymphocytes and splenic lymphocyte. The results showed that GPSL could significantly enhance T and B lymphocytes proliferation singly or synergistically with PHA and LPS and the efficacy were superior to those of gypenosides (GPS) and blank liposome (BL) at most of concentrations. In vivo, three hundred and fifty 14-day-old chickens were assigned to 7 groups randomly and vaccinated with Newcastle disease (ND) vaccine. Simultaneously, the chickens in experimental groups were, respectively, oral administration with the GPSL at three doses, GPS and BL. The results showed that GPSL could significantly enhance lymphocyte proliferation, increase antibody titer, and promote cytokine secretion in vitro and in vivo, moreover, the adjuvant activity of GPSL was better than those of GPS and BL. These indicated that formulations of GPS and liposome can further enhance the immune response against ND vaccine compared with the adjuvant alone.

Optimization on preparation condition of propolis flavonoids liposome by response surface methodology and research of its immunoenhancement activity.

The aim of this study is to prepare propolis flavonoids liposome (PFL) and optimize the preparation condition and to investigate further whether liposome could promote the immunoenhancement activity of propolis flavonoids (PF). PFL was prepared with ethanol injection method, and the preparation conditions of PFL were optimized with response surface methodology (RSM). Moreover, the immunoenhancement activity of PFL and PF in vitro was determined. The result showed that the optimal preparation conditions for PFL by response surface methodology were as follows: ratio of lipid to drug (w/w) 9.6 : 1, ratio of soybean phospholipid to cholesterol (w/w) 8.5 : 1, and speed of injection 0.8 mL·min(-1). Under these conditions, the experimental encapsulation efficiency of PFL was 91.67 ± 0.21%, which was close to the predicted value. Therefore, the optimized preparation condition is very reliable. Moreover, the results indicated that PFL could not only significantly promote lymphocytes proliferation singly or synergistically with PHA, but also increase expression level of IL-2 and IFN-γ mRNA. These indicated that liposome could significantly improve the immunoenhancement activity of PF. PFL demonstrates the significant immunoenhancement activity, which provides the theoretical basis for the further experiment in vivo.

Effect of epimedium polysaccharide-propolis flavone immunopotentiator on immunosuppression induced by cyclophosphamide in chickens.

Two hundred and fifty 11-day-old chickens were randomly assigned into 5 groups and except normal control group injected with cyclophosphamide once a day for 3 successive days. At day-14-old, all chickens were vaccinated with Newcastle disease vaccine. At the same time of the first vaccination, the chickens in three experimental groups were injected respectively with epimedium polysaccharide-propolis flavone immunopotentiator (EPI) at three dosages, once a day for 3 successive days. On days 7, 14, 21 and 28 after the first vaccination, the serum antibody titer and IgG, IgM, IFN-γ and IL-6 concentrations, peripheral lymphocyte proliferation, including immune organ index on day 28, were measured. The results demonstrated that EPI at high and medium doses could significantly enhance antibody titer and IgG, IgM, IFN-γ and IL-6 concentrations, promote lymphocyte proliferation and enlarge immune organ index as compared with model control group. This indicated that EPI could effectively resist the immunosuppression.

Optimization on preparation condition of epimedium polysaccharide liposome and evaluation of its adjuvant activity.

The aim of this strategy was to investigate whether the adjuvant activity of epimedium polysaccharide (EPS) could be further enhanced after encapsulated with liposome. In preparation of EPS liposome (EPSL) test, an orthogonal L(9) (3(4)) test design was used to optimize the preparation condition of EPSL. In adjuvant activity test, 350 14-day-old chickens were randomly assigned to 7 groups and vaccinated with Newcastle disease (ND) vaccine. Simultaneously, the chickens in experimental groups were injected with EPSL at three doses, EPS and blank liposome, respectively. The activity of lymphocytes proliferation, titer of serum antibody and concentrations of cytokines were determined. Results showed that the optimal preparation condition of EPSL was that ratio of drug to lipid, ratio of soybean phospholipid to cholesterol, ultrasonic time, and water bath temperature were 1:30, 4:1, 10 min and 40°C, respectively. EPSL could significantly enhance the immune response of ND vaccine and promote cytokines secretion, and its high dose possessed the best efficacy. These findings indicated that liposome encapsulation could significantly improve the adjuvant activity of EPS.