•Garlic polysaccharide (GPS) was modified in selenylation respectively by four methods.•Contents and yields of selenium and immune-enhancing activities of sGPSs were compared.•Selenylation modification can significantly enhance immune-enhancing activity of GPS.•NA-SS method presented the highest selenylation efficiency and was the best method.•sGPS5 displayed strongest activity and optimal modification conditions were gotten.Garlic polysaccharide (GPS) was modified in selenylation respectively by nitric acid-sodium selenite (NA-SS), glacial acetic acid-selenous acid (GA-SA), glacial acetic acid-sodium selenite (GA-SS) and selenium oxychloride (SOC) methods each under nine modification conditions of L9(34) orthogonal design and each to obtain nine selenizing GPSs (sGPSs). Their structures were identified, yields and selenium contents were determined, selenium yields were calculated, and the immune-enhancing activities of four sGPSs with higher selenium yields were compared taking unmodified GPS as control. The results showed that among four methods the selenylation efficiency of NA-SS method were the highest, the activity of sGPS5 was the strongest and significantly stronger than that of unmodified GPS. This indicates that selenylation modification can significantly enhance the immune-enhancing activity of GPS, NA-SS method is the best method and the optimal conditions are 0.8:1 weight ratio of sodium selenite to GPS, reaction temperature of 70 °C and reaction time of 10 h.

•One acidic Epimedium polysaccharide was obtained by chromatographic method.•EPS-1 may be composed of GalpA, Galp and Rhap residues in a ratio of 12:1:1.•EPS-1 can promote the proliferation and cytokine secretion of chicken T cells.•EPS-1 can also increase the immune abilities of immature chBM-DCs.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.

•Nine selenizing lily polysaccharides, sLP1–sLP9, were prepared.•sLPs’ immune-enhancing activities were compared taking unmodified LP as control.•Selenylation modification can significantly enhance LP's immune-enhancing activity.•sLP6 presents the best effect and can be as a new polysaccharide immunopotentiator.Lily polysaccharide (LP) was extracted, purified and selenizingly modified by HNO3–Na2SeO3 method according to L9(34) orthogonal design. Nine selenizing LPs, sLP1–sLP9, were obtained and their immune-enhancing activities were compared taking unmodified LP as control. The results in vitro test showed that sLP6 presented the strongest activity in promoting lymphocytes proliferation in single and synergetic with PHA, and the relative expression level of IL-2, IL-6 and IFN-γ mRNA of chicken peripheral lymphocytes. The results in vivo test showed that sLP6 could promote lymphocytes proliferation and enhance the serum antibody titers and serum IL-2, IL-6, IFN-γ contents more significantly than LP in chickens vaccinated with Newcastle Disease (ND) vaccine. These results indicate that polysaccharide selenizing can significantly enhance the immune-enhancing activity of LP and the optimal modification conditions are 400 mg of Na2SeO3 per 500 mg of LP, the reaction temperature of 70  °C and the reaction time of 6 h.

•We studied the chemical constituents of Euphorbia hirta and obtained thirty-one compounds.•The structures of compounds were elucidated by spectroscopic methods.•Five compounds were firstly isolated from Euphorbia, and seventeen compounds were firstly obtained from this species.•The obtained constituents showed relationships among the Euphorbia plants.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.

•Nine selenizing lycium barbarum polysaccharide, sLBP1–sLBP9, were prepared.•sLBPs’ antioxidant activities were compared taking non-modified LBP as control.•Selenylation modification could significantly raise LBP's antioxidant activity.•sLBP6 presented best effect and could be as a candidate drug of polysaccharide antioxidants.•Optimal modification conditions were 500 mg LBP adding 400 mg Na2SeO3 and reacting 6 h at 70 °C.Lycium barbarum polysaccharide (LBP) was modified by HNO3–Na2SeO3 method according to L9(34) orthogonal design to obtain nine selenizing LBPs (sLBPs), sLBP1–sLBP9. Their antioxidant activities in vitro were compared by free radical-scavenging test. sLBP6, sLBP8 and sLBP9 presented stronger activity. In vivo test, 14-day-old chickens were injected respectively with sLBP6, sLBP8 and sLBP9 taking LBP as control, and serum GSH-Px and SOD activities and MDA content were determined. The results showed that three sLBPs could significantly enhance GSH-Px and SOD activities and decrease MDA content. The actions of sLBPs were significantly stronger than that of unmodified LBP. These results indicated that selenylation modification could significantly enhance the antioxidant activities of LBP, sLBP6 possessed the best efficacy and could be exploited into an antioxidant. The optimal modification conditions were 400 mg of sodium selenite for 500 mg of LBP, reaction temperature of 70 °C and reaction time of 6 h.

•We studied the isolation of Cyclocarya paliurus and obtained sixteen compounds.•The structures of these compounds were elucidated by spectroscopic methods.•Cyclocarioside K was a new compound and five other compounds were firstly obtained from Cyclocarya genus.•C. paliurus has close relationship with some species of Juglandaceae family.A novel triterpenoid, cyclocarioside K, together with fifteen known compounds, including seven triterpenoids (2–8), five flavonoids (9–13), three phenolic derivatives (14–16), was obtained from the leaves of Cyclocarya paliurus (Batal.) Iljinsk (Juglandaceae). Their structures were elucidated by spectroscopic methods, including two-dimensional NMR experiments (HSQC DEPT and HMBC). The chemotaxonomic relationships were also discussed. As a result, 3, 4-secodammarane triterpenoids and kaempferol glycosides may represent phytochemical fingerprints for C. paliurus, and C. paliurus has close relationships with other species of Juglandaceae.

•We studied the chemical constituents of the fruit of Zizyphus jujuba Mill. var. spinosa and obtained twenty-one compounds.•The structures of compounds were elucidated by spectroscopic methods.•Compounds 11–15 and 19–21 were firstly obtained from genus Zizyphus.•The obtained constituents showed the relationships among the plants of Rhamnaceae family.Twenty-one compounds, including ten triterpenoids (1–10), two sterols (11 and 12), six flavonoids (13–18), and three cerebrosides (19–21) were isolated from the fruit of Zizyphus jujuba Mill. var. spinosa (Bunge) Hu ex. H. F. Chou. The structures were elucidated by spectroscopic methods and by comparison of their reported spectral data. These compounds have shown the relationship between this plant and other species from the Rhamnaceae family.

•We studied the isolation of Ilex urceolatus and obtained eighteen compounds.•The structures of these compounds were elucidated by spectroscopic methods.•Compounds 14–18 were firstly isolated from genus Ilex.•Compounds 1–13 were obtained from I. urceolatus for the first time.•I. urceolatus had close relationships with other species from Ilex.Eighteen compounds, including four hemiterpene glycosides (1–4), three triterpenoid saponins (5–7), four triterpenes (8–11), five sterols (12-16) and two monoterpene glucosides (17 and 18), were isolated from the leaves of Ilex urceolatus C. B. Shang, K. S. Tang et D. Q. Du, which was identified as a new species belonging to the genus Ilex. Among them, compounds 14–18 were firstly isolated from the genus Ilex, others were obtained from I. urceolatus for the first time. This work represented the initially phytochemical study on this plant. The isolated compounds have significant chemotaxonomic characteristics with the other species from this genus.

•sTPS70c and sCPPStp demonstrated better effect in vitro.•Sulfation modification improved the immune-enhancing activity of TPS and CPPS in vivo.•sTPS70c at medium dosage presented the best efficacy.•sTPS70c would be expected as a component of new-type immunopotentiator.Based on our previous research, four sulfated polysaccharide (sPSs) from Tremella and Condonpsis pilosula, sTPStp, sTPS70c, sCPPStp and sCPPS50c, were prepared and their effects on splenic lymphocytes proliferation in vitro and the immune response of ND vaccine in chicken were compared taking the unmodified polysaccharide (uPS) TPStp as control. The results showed that four sPSs could significantly or numerically stimulate splenic lymphocyte proliferation singly or synergistically with LPS in vitro, sTPS70c and sCPPStp demonstrated better effect; promote peripheral lymphocytes proliferation and enhance serum HI antibody titer in chickens vaccinated with ND vaccine, the actions of sPSs were stronger than that of uPS, and sTPS70c at medium dosage presented the best efficacy. These indicated that sulfation modification could improve the immune-enhancing activity of TPS and CPPS, sTPS70c possessed the strongest activity and would be expected as a component of new-type immunopotentiator.

Chinese angelica polysaccharide (CAP) was extracted by water decoction and ethanol precipitation, purified through eliminating protein by Sevage method and column chromatography of Sephadex G-200, then selenizingly modified by nitric acid–sodium selenite method according to L9(34) orthogonal design of three-factors, the usage amount of sodium selenite, reaction temperature and reaction time, at three level to obtain nine selenizing CAPs, sCAP1–sCAP9. Their effects on chicken peripheral lymphocytes proliferation in vitro were compared by MTT assay taking the non-modified CAP as control. The results showed that selenylation modification could significantly enhance the immune-enhancing activity of CAP, sCAP2 presented best effect and the optimal modification conditions were 200 mg of sodium selenite for 500 mg of CAP, the reaction temperature of 70 °C and the reaction time of 6 h.Highlights► Nine selenizing Chinese angelica polysaccharide, sCAP1–sCAP9, were prepared. ► sCAPs’ immune-enhancing activities were compared taking non-modified CAP as control. ► Selenylation modification could significantly raise CAP's immune-enhancing activity. ► sCAP2 presented best effect and the optimal modification conditions were defined.

The preparation conditions of epimedium polysaccharide-propolis flavone liposome (EPL) were optimized by response surface methodology taking entrapment rates of epimedium polysaccharide and propolis flavone as indexes. The immunoenhancement of EPL prepared with optimized condition was determined taking epimedium polysaccharide-propolis flavone suspension (EPS) and epimedium polysaccharide-propolis flavone watery solution (EPW) as control. The results showed that the optimized preparation condition was as follows: the ratio of drug to lipid was 14:1, the ratio of soybean phospholipid to cholesterol was 6:1, and the ultrasonic time was 19 min. EPL could significantly promote the proliferation of T and B lymphocytes singly or synergistically with PHA or LPS, mRNA expression of IL-2 and IL-6 and secretion of IgG and IgM as compared with EPS and EPW. These results indicated that liposome could significantly improve the immunoenhancement of epimedium polysaccharide-propolis flavone immunopotentiator (EPI) and would be as the suitable dosage form of EPI.Highlights► Preparation conditions of EPL were optimized by response surface method. ► The optimal preparation condition for EPL was set up. ► EPL could significantly promote T and B lymphocytes proliferation. ► EPL could promote mRNA expression of IL-2 and IL-6 and secretion of IgG and IgM.

•Three selenizing Chinese angelica polysaccharide, sCAP2, sCAP6 and sCAP8, were prepared.•Three sCAPs’ immune-enhancing activities were compared taking non-modified CAP as control.•Selenylation modification could significantly raise CAP's immune-enhancing activity.•sCAP2 presented best effect and would be as a component drug of new-type immunoenhancer.On the basis of previous test that selenizing Chinese angelica polysaccharides (sCAPs) with stronger immune-enhancing activity in vitro were picked out, the immune-enhancing activity in vivo of three sCAPs, sCAP2, sCAP6 and sCAP8, at high and low dosage were compared taking the unmodified Chinese angelica polysaccharide (CAP) as control by determination of peripheral lymphocyte proliferation, serum antibody titer, IFN-γ and IL-6 contents in chicken vaccinated with Newcastle Disease vaccine. The results showed that three sCAPs at suitable dosage could significantly promote lymphocyte proliferation, enhance serum antibody titer, IFN-γ and IL-6 contents as compared with unmodified CAP, sCAP2 at low dosage possessed the strongest action. These results indicated that selenylation modification could significantly enhance the immune-enhancing activity of CAP, sCAP2 possessed the best efficacy and would be as a component drug of new-type immunoenhancer.

The immunoenhancement of compound polysaccharides, APS–sEPS composed with astragalus polysaccharide (APS) and sulfated epimedium polysaccharide (sEPS), was observed in immunosuppressed model chicken induced by cyclophosphamide (Cy). 11-day-old chickens were injected with Cy once a day for three successive days except vaccine control group. At day-14-old, all chickens were vaccinated with ND vaccine, and in experimental groups simultaneously administrated with APS–sEPS at three dosages, APS and sEPS once a day for three successive days. On days 7, 14, 21 and 28 after the administration, the peripheral T-lymphocyte proliferation, serum antibody titers, IFN-γ, IL-2, IgG and IgM were determined. The results displayed that APS–sEPS could overcome Cy-induced immunosuppression, significantly promote T-lymphocyte proliferation and raised serum antibody titers, IFN-γ, IL-2, IgG and IgM levels, its high and medium doses were superior to single APS or sEPS. This demonstrated that APS and sEPS could synergistically resist the immunosuppression and APS–sEPS was an effective immunopotentiator.Highlights► The immunoenhancement of APS–sEPS was observed in immunosuppressed model chicken. ► APS–sEPS could promote T-lymphocyte proliferation and raise serum antibody titers. ► APS–sEPS could enhance serum IFN-γ, IL-2, IgG and IgM level. ► APS and sEPS could synergistically resist the immunosuppression. ► APS–sEPS was an effective immunopotentiator.

Total Auricularia auricula polysaccharide (AAPt) was prepared by extracting and removing the proteins. Column chromatography was used to further graded it into AAP1 and AAP2. Three AAPs were modified by chlorosulfonic acid–pyridine method to obtain three sulfated AAPs (sAAPs), sAAPt, sAAP1 and sAAP2, respectively. Three sAAPs and Newcastle disease virus (NDV) were added into cultivation system of chicken embryo fibroblast (CEF) in three manners, pre-, post- and simultaneous-adding polysaccharide with NDV respectively, taking three non-modified AAPs as control. Their anti-viral activities were compared by MTT method. The results showed that sAAPs and AAPs at a certain concentration could significantly inhibit the cellular infectivity of NDV in three manners. The effects of sAAPs were better than that of AAPs. It indicated that sulfated modification could enhance the antiviral activity of AAP. sAAP1 and sAAPt possessed stronger activity and would be as the component of a new-type antiviral drug.Highlights► Three sulfated A. auricula polysaccharides, sAAPt, sAAP1 and sAAP2, were prepared. ►The antiviral activities of sAAPs were compared taking the non-sulfated AAPs as control. ► Sulfation modification could enhance the antiviral activity of AAP. ► sAAP1 and sAAPt were better and could be used as component of a new-type antiviral drug.

The crude total Auricularia auricula polysaccharide (AAPct) was extracted by water decoction and ethanol precipitation, protein was removed to obtain total A. auricula polysaccharide (AAPt), then was graded into AAP1 and AAP2 through column chromatography. sAAPt, sAAP1 and sAAP2 were prepared by chlorosulfonic acid-pyridine method. In vitro test, the effects of sAAPt, sAAP1, sAAP2, AAPt, AAP1 and AAP2, on chicken peripheral lymphocytes proliferation were compared. The results showed that sAAPt and sAAP1 demonstrated better effect. In vivo test, 14-day-old chickens were injected respectively with sAAPt, sAAP1, AAPt and AAP1 at the first vaccination of ND vaccine, once a day for three days. On days 7, 14, 21 and 28 after the first vaccination, the peripheral lymphocytes proliferation and antibody titer were determined. The results indicated that sAAPt possessed the best efficacy and would be expected to be used as a component of a new-type immunopotentiator.Highlights► Three sulfated A. auricula polysaccharides (sAAPt, sAAP1 and sAAP2) were prepared. ► Immune-enhancing activities of sAAPs were compared taking the non-sulfated AAP as control. ► Sulfation modification could significantly enhance the immune-enhancing activity of AAP. ► sAAPt is the best and could be used as component of a new-type immunopotentiator.

To evaluate the immune activation and reactive oxygen species scavenging activity of Cordyceps militaris polysaccharides (CMP) in vivo, 90 male BALB/c mice were randomly divided into six groups. The mice in the three experimental groups were given cyclophosphamide at 80 mg/kg/d via intraperitoneal injection and 17.5, 35, or 70 mg/kg body weight CMP via gavage. The lymphocyte proliferation, phagocytic index, and biochemical parameters were measured. The results show that the administration of CMP was able to overcome the CY-induced immunosuppression, significantly increased the spleen and thymus indices, and enhanced the spleen lymphocyte activity and macrophage function. CMP can also improve the antioxidation activity in immunosuppressed mice, significantly increase the superoxidase dismutase, catalase, and glutathione peroxidase levels and the total antioxidant capacity, and decrease the malondialdehyde levels in vivo.Highlights► Immunosuppressed mice was administrated Cordyceps militaris polysaccharides (CMP). ► CMP can overcome the cyclophosphamide-induced immunosuppression. ► Enhancing spleen lymphocyte activity and macrophage function. ► Raising the SOD, CAT, GSH-Px and TAOC level and the spleen and thymus indices. ► Decreasing MDA level.

The total Siberian solomonseal rhizome polysaccharide (SRPSt) and four fractional SRPS30, SRPS50, SRPS70 and SRPS80, were extracted by one-step or stepwise ethanol precipitation method. In test in vitro, their effects on chicken peripheral lymphocytes proliferation were compared. The result showed that SRPS70 and SRPS50 presented better effect. In test in vivo, 14-day-old chickens were injected, respectively, with SRPS50, SRPS70 and SRPSt, at the first vaccination of ND vaccine, once a day for three successive days. On days 7, 14, 21 and 28 after the first vaccination, the peripheral lymphocytes proliferation and antibody titer were determined. The result indicated that lymphocyte proliferation and antibody titers in SRPS50 group at all time points were the highest, significantly higher than those of non-adjuvant control group and most of another two SRPSs groups. These results confirmed that SRPS50 possessed the strongest action and was the immune-enhancing active site of SRPS.

In order to investigate the possibility of sulfating modification in improving the anti-viral activity of lentinan (LNT), three kinds of sulfated LNTs (sLNT), sLNT1, sLNT2 and sLNT3 with the degree of substitution of 0.69, 0.98 and 1.37 were prepared by chlorosulfonic acid–pyridine method with three modification condition, respectively. The sLNTs and avain infectious bronchitis virus (IBV) were added into chicken embryo fibroblast (CEF) in three manners, pre-adding polysaccharide, post-adding polysaccharide and mixed adding polysaccharide with IBV, respectively, taking non-modified LNT as control. The anti-viral activities of three sLNTs were compared by MTT method. The results showed that all of sLNTs and LNT at a certain concentration could significantly inhibit IBV growth in three manners. The effects of modified sLNTs were better than that of non-modified LNT. It indicated that sulfated modification could enhance the anti-viral activity of LNT.

In order to select the sulfated polysaccharide prescription (sPSP) with the best immunoenhancement, 270 14-day-old chickens were averagely divided into nine groups and vaccinated with ND-IV vaccine. The chickens in seven experimental groups were intramuscularly injected with three sulfated polysaccharides and their four prescriptions (sPSPs). The serum antibody and peripheral lymphocyte proliferation were measured. On day 23 after vaccination, all chickens except for Negative control group were challenged with Newcastle disease virus. The morbidity, mortality and immune protective rate were calculated and their serum antibody titers were determined. The results showed that all sulfated polysaccharides and their prescriptions could obviously or significantly enhance the immune response of ND vaccine in vaccinated chicken and increase the immune protective rate in challenged chicken. sPSP1 possessed the best efficacy, stronger than its components as well, and would be expected as a candidate of new-type immune adjuvant.

•The selenizing Chinese angelica polysaccharide(sCAP2)and selenizing garlic polysaccharide (sGPS6) were prepared.•The selenizing polysaccharides and non-selenizing polysaccharides on murine macrophages function were firstly compared.•Selenylation modification can significantly enhance immune-enhancing activity of polysaccharide.•sCAP2 displayed the strongest activity and would be as a new immunopotentiator.The effects of two selenizing polysaccharides (sCAP2 and sGPS6) on immune function of murine peritoneal macrophages taking two non-selenizing polysaccharides (CAP and GPS) and modifier Na2SeO3 as control. In vitro test, the changes of selenizing polysaccharides, non-selenizing polysaccharides and Na2SeO3 on murine macrophages function were evaluated by phagocytosis and nitric oxide (NO) secretion tests. In vivo test, the mice were injected respectively with 0.2, 0.4 and 0.6 mg of sCAP2, sGPS6, CAP and GPS, or Na2SeO3 80 μg or normal saline 0.4 mL. The peritoneal macrophages were collected and cultured to determine the contents of TNF-α, IL-6 and IL-10 in supernatants by enzyme-linked immunosorbent assay. The results showed that sCAP2 and sGPS6 could significantly promote the phagocytosis and secretion of NO and three cytokines of macrophages in comparison with CAP and GPS. sCAP2 possessed the strongest activity. This indicates that selenylation modification can further improve the immune-enhancing activity of polysaccharide, and sCAP2 could be as a new immunopotentiator.

•Codonopsis pilosula polysaccharide (CP) was modified to obtain sulfated CP (sCP).•Antioxidant activities of sCP and CP were compared by free radical scavenging test.•Hepatoprotective activities of sCP and CP were compared in hepatic injury model mice.•sCP possessed better antioxidative and hepatoprotective activities as compared with CP.•Sulfation modification can enhance the antioxidative activity of CP.Codonopsis pilosula polysaccharide (CP) was extracted, purified and modified by chlorosulfonic acid-pyridine method to obtain a sulfated CP (sCP). Their antioxidative activities in vitro were compared through the free radical-scavenging test. The results demonstrated that the scavenging capabilities of sCP were significantly stronger than those of CP. In vivo test, the mice hepatic injury model was prepared by BCG/LPS method, then administrated respectively with sCP and CP at three dosages, the biochemical indexes in serum, antioxidative indexes in liver homogenate and histopathological change in liver of the mice were compared. The results showed that in high (200 mg/kg) and middle (150 mg/kg) dosages of sCP groups, the contents of ALT, AST and TNF-α in serum and MDA in liver homogenate were significantly lower than those in the model group and numerically lower than those in the CP groups, the activities of SOD and GSH-Px in liver homogenate were significantly higher than those in the model group and numerically higher than those in the CP groups. In the model group there were obvious pathological changes in the liver, while in the sCP groups were near normal. These results indicate that sCP and CP possess antioxidative activity in vitro and in vivo, the activity of sCP is stronger than that of CP and sulfation modification can enhance the antioxidative and hepatoprotective activities of Codonopsis pilosula polysaccharide.

ObjectiveIn order to select the component drug in promoting blood circulation and removing blood stasis of Chinese herbal medicinal formula for dairy cow mastitis.Methods25 healthy rabbits were allocated randomly into five equal groups. The rabbits in four experimental groups were administered with decoctions of giant knotweed rhizome (GKR, rhizoma polygoni cuspidati), safflower (SF, flos carthami), red sage root (RSR, radix salviae miltiorrhizae) and chuanxiong rhizome (CXR, rhizoma Chuanxiong) by gastrogavage, respectively, in control group, physiological saline, once a day for seven successive days. After the last administration, all rabbits were intravenously injected with 10% macromolecular dextran to induce blood stasis. The blood samples of all rabbits were collected before the first administration, at 2 h after the last administration and 1 h after injection of dextran, respectively for determination of hemorheologic parameters by MVIS-2035 hemorheology auto-analyzing system.ResultsThe results showed that all of four kinds of herbs presented different degree of activating blood flow and removing blood stasis.ConclusionRed sage root was the best especially in resisting blood stasis induced by dextran, and would be selected as main component drug of the prescription for dairy cow mastitis.