Hirsutella sinensis mycelium polysaccharides attenuate the TGF-ß1-induced epithelial-mesenchymal transition in human intrahepatic bile duct epithelial cells.

Hirsutella sinensis is the anamorph of Ophiocordyceps sinensis, and its mycelia has been used to effectively treat a variety of hepatobiliary diseases in clinical practice. In the present study, we performed a systematic study on the composition and structure of its polysaccharides, and then employed a TGF-ß1-induced human intrahepatic bile duct epithelial cell-epithelial-mesenchymal transition (HIBEC-EMT) model to investigate their effects on treating primary biliary cholangitis (PBC) based on hepatic bile duct fibrosis. Four polysaccharide fractions were obtained from H. sinensis mycelia by hot-water extraction, DEAE-cellulose column and gradient ethanol precipitation separation. HSWP-1a was an α-(1,4)-D-glucan; HSWP-1b and HSWP-1d mainly consisted of mannoglucans with a backbone composed of 1,4-linked α-D-Glcp and 1,4,6-linked α-D-Manp residues branched at O-6 of the 1,4-linked α-D-Glcp with a 1-linked α-D-Glcp as a side chain; and HSWP-1c mainly contained galactomannoglucans. These polysaccharide fractions protected HIBECs from a TGF-ß1-induced EMT, according to HIBEC morphological changes, cell viability, decreased E-cadherin and ZO-1 expression, and increased vimentin and collagen I expression. Furthermore, the effects of the polysaccharides might be mediated by inhibiting the activation of the TGF-ß/Smad signaling pathway, which attenuated hepatic bile duct fibrosis and potential PBC effects.

Bioinformatics analysis and in vivo validation study of Ophiocordyceps sinensis (Berk.)G.H.Sungetal against lung adenocarcinoma.

ETHNOPHARMACOLOGICAL RELEVANCE: Lung adenocarcinoma (LUAD) is one of the main types of lung cancer. Ophiocordyceps sinensis has many potentially useful pharmacologic features, such as lung protection, and both anti-inflammatory and antioxidant activities. AIM OF THE STUDY: This study was conducted to investigate-using bioinformatics and in vivo experimental validation-the possible role of O. sinensis against LUAD. MATERIALS AND METHODS: We obtained important targets of O. sinensis for the treatment of LUAD using network pharmacology techniques and deep mining of the TCGA database, and validated them by molecular docking techniques and in vivo experiments. RESULTS: Through bioinformatics analysis and research, we screened BRCA1 and CCNE1 as important biomarkers for LUAD and as core targets of O. sinensis against LUAD. The non-small cell lung cancer signaling pathway, PI3K-Akt signaling pathway, and HIF-1 signaling pathway are potentially important pathways of O. sinensis against LUAD. The molecular docking results showed good binding between the active components in O. sinensis and the two core targets, and the in vivo experimental validation results indicated that O. sinensis had good inhibitory effects in the Lewis lung cancer (LLC) model. CONCLUSIONS: BRCA1 and CCNE1 are crucial biomarkers for LUAD and are important targets for O. sinensis to exert anti-LUAD effects.

Structure and immunomodulatory activity of a water-soluble α-glucan from Hirsutella sinensis mycelia.

Hirsutella sinensis, the anamorph of Ophiocordyceps sinensis (syn. Cordyceps sinensis), is a great substitute for precious and rare wild Cordyceps sinensis to effectively treat a variety of lung and kidney diseases. In this study, an α-glucan (named as HSWP-2a) was obtained by hot water extraction, DEAE-cellulose separation, and Sepharose CL-6B purification from H. sinensis mycelia. Different from known α-glucans, HSWP-2a is an α-(1 â†’ 4)-D-glucan that branched at O-6, O-3, or O-2 with a terminal 1-linked α-D-Glcp as side chain, with an average molecular weight of 870.70 kDa. Immunological tests showed that HSWP-2a could remarkably enhance the phagocytosis of macrophages and increase the production of NO, IL-1ß, IL-6, and TNF-α, via activating the p38, JNK, and NF-κB signaling pathways. Moreover, HSWP-2a could significantly promote splenic lymphocyte proliferation. Taken together, HSWP-2a may be potentially utilized as a natural immunomodulatory agent.

Neurotoxicity of ß-HgS differs from environmental mercury pollutants (MeHgCl and HgCl2) in Neuro-2a cell.

ß-HgS, differing from environmental mercury pollutants (MeHgCl and HgCl2) in chemical form, is used as traditional medicine in Asian countries for thousands of years. In this study, Neuro-2a cells were exposed to ß-HgS, MeHgCl and HgCl2 (5 µM) for 6-24 h. The cell viability of ß-HgS was higher than MeHgCl with 25.9% and 72.4% in 12 h and 24 h respectively. As the incubation time increased, MeHgCl had obvious damage to cell morphology, decreased the ratio of Bcl-2 and Bak and increased the expressions of TNF-α, IL-6 and IL-1ß significantly. Furthermore, the expressions of IL-1ß and IL-6 in HgCl2 group were increased significantly in 6 h and 24 h. The apoptotic rates in MeHgCl and HgCl2 group were respectively higher than ß-HgS with 32.2% and 7.30% in 24 h. Our findings indicate that ß-HgS is much less neurotoxicity than MeHgCl and HgCl2 in Neuro-2a cells.

Tobacco Mosaic Viral Nanoparticle Inhibited Osteoclastogenesis Through Inhibiting mTOR/AKT Signaling.

INTRODUCTION: Tobacco mosaic virus-based nanoparticles (TMV VNPs) were previously shown to promote osteogenic differentiation in vitro. This study aims to investigate whether and how TMV VNPs impact on osteoclastogenesis in vitro and bone injury healing in vivo. METHODS: Raw264.7 cells were cultured in osteoclastogenic medium in culture plates coated with or without TMV and TMV-RGD1 VNPs, followed by TRAP staining, RT-qPCR and WB assessing expression of osteoclastogenic marker genes, and immunofluorescence assessing NF-κB activation. TMV and TMV-RGD1-modified hyaluronic acid hydrogel were used to treat mouse tibial bone injury. Bone injury healing was checked by micro-CT and Masson staining. RESULTS: TMV and TMV-RGD1 VNPs significantly inhibited osteoclast differentiation and downregulated the expression of osteoclastogenic marker genes Ctr, Ctsk, Mmp-9, Rank, and Trap. Moreover, TMV and TMV-RGD1 VNPs inhibited NF-κB p65 phosphorylation and nuclear translocation, as well as activation of mTOR/AKT signaling pathway. TMV and TMV-RGD1-modified HA hydrogel strongly promoted mouse tibial bone injury with increased bone mass compared to plain HA hydrogel. The amount of osteoclasts was significantly reduced in TMV and TMV-RGD1 treated mice. TMV-RGD1 was more effective than TMV in inhibiting osteoclast differentiation and promoting bone injury repair. DISCUSSION: These data demonstrated the great potential of TMV VNPs to be developed into biomaterial for bone injury repair or replacement.

Ma-Nuo-Xi decoction has an immunostimulating effect in cyclophosphamide-immunosuppressed mice.

BACKGROUND: Ma-Nuo-Xi decoction (MNXD), as well as its hundreds of derivative preparations, has been used in Tibetan medicine since the 14th century. MNXD is in accordance with the theory of treatment determination based on syndrome differentiation. This study aimed to compare the effect of the auxiliary MNXD prescription (MNXD-AD) with that of the basic MNXD prescription (MNXD-BD) on the immunostimulating activity of MNXD. METHODS: The immunopotentiation of MNXD, MNXD-BD, and MNXD-AD was evaluated using a cyclophosphamide (CTX)-immunosuppressed mouse model. Their influences on non-specific and specific immunity were evaluated using immune organ indexes, peripheral white blood cell (WBC) count, red blood cell (RBC) count, platelet count, phagocytosis, macrophage-secreted nitric oxide (NO) and cytokines, natural killer (NK) cytotoxic activity, lymphocyte proliferation, serum cytokines, splenic T-lymphocyte subpopulations, and quantitative hemolysis of sheep red blood cell (QHS SRBC) assays. RESULTS: MNXD, MNXD-BD, and MNXD-AD increased the spleen and thymus indexes, as well as the peripheral WBC, RBC, and platelet counts. They also promoted phagocytosis, NO and cytokine secretion from macrophages, NK cytotoxic activity, and lymphocyte proliferation, and also raised the CD4+ /CD8+ T-cell ratio, serum cytokine concentrations, and haemolysin formation in CTX-treated immunosuppressed mice. Compared with MNXD-BD and MNXD-AD, MNXD was superior in restoring the phagocytic index, concanavalin A (ConA)-induced T-lymphocyte proliferation, NO secretion from macrophages, and haemolysin formation, as well as the levels of interleukin 1 beta (IL-1ß), and serum interleukin-2 (IL-2) and interferon gamma (INF-γ). CONCLUSIONS: MNXD, MNXD-BD, and MNXD-AD have excellent immunostimulating and myelosuppression-restoring activities on CTX-immunosuppressed mice. Among them, MNXD-AD might be an immunomodulator, which may happen to be in line with the clinical experience of Tibetan medicine physicians of using it to promote the efficacy of MNXD-BD.

Effects of ß-HgS on cell viability and intracellular oxidative stress in PC-12 cells.

Traditional Tibetan medicines containing ß-HgS have been used to treat chronic ailments for thousands of years. However, there has recently been speculation regarding the safety of these medicines due to their high mercury content. Although the toxic effect of ß-HgS has been previously investigated in vivo, the mechanism underlying the toxicity of this compound remains unclear. In this study, we investigate the mechanism of ß-HgS cytotoxicity via experiments performed on rat adrenal gland tumor cells (PC-12). Specifically, we analyze the viability and intracellular oxidative stress state of PC-12 cells treated with varying concentrations of ß-HgS. For comparison purposes, the effects of MeHgCl and HgCl2, two Hg-based compounds, on ROS generation and MDA, GSH/GSSG, Nrf2, NQO-1, and HO-1 levels are also determined. It should be noted that we used the small-molecule thiols of cell culture medium, such as cysteine, to increase the solubility of ß-HgS and prepare a ß-HgS solution to treat PC-12 cells. The obtained results show that ß-HgS inhibits cell viability at concentrations of 200-1000 ng Hg mL-1 (48 h treatment). In the concentration range of 200-600 ng Hg mL-1 (24 h treatment), the inhibitory effect of ß-HgS is stronger than that of MeHgCl; however, this trend is reversed at higher concentrations (800-1000 ng mL-1) and longer exposure times (48 h). Moreover, ß-HgS significantly promotes MDA, but has no appreciable influence on cell apoptosis and ROS generation in PC-12 cells, which suggests that its inhibitory effect on cell viability might be related to the stimulation of ROS-independent oxidative stress. Notably, ß-HgS and HgCl2 significantly increase the GSH content, GSH/GSSG ratio, NQO-1 mRNA expression, and HO-1 protein expression in PC-12 cells, indicating that the antioxidant protection against these compounds is triggered by Nrf2 activation. HPLC-AFS analysis shows that in ß-HgS and HgCl2 solutions, mercury exists in the same form of Hg2+, but the cytotoxicity of the former is greater. This is probably due to the additional oxidative damage induced by the S2- ion in ß-HgS. In conclusion, ß-HgS induces ROS-independent oxidative stress in PC-12 cells, and thus, is obviously cytotoxic. At the same time, it promotes the antioxidant capacity of cells by activating the Nrf2 pathway.

Immunostimulating effect of polysaccharides isolated from Ma-Nuo-Xi decoction in cyclophosphamide-immunosuppressed mice.

Ma-Nuo-Xi Decoction (MNXD) is well-known in Tibetan medicine to be in line with the theory of treatment determination based on syndrome differentiation. However, the components responsible for its immunomodulating effect are unknown. In this study, three polysaccharide components-MNXD-P, MNXD-BD-P, and MNXD-AD-P-were isolated from MNXD and its basic and auxiliary prescription decoctions, of which MNXD-BD-P is composed of ß-(1,4)-d-glucan and RG-I pectin, MNXD-AD-P contains mainly α-(1,4)-d-glucan and some amount of arabinogalactan and/or arabinorhamnogalactan, and MNXD-P contains components of both MNXD-BD-P and MNXD-AD-P. And treatment with these polysaccharides could significantly improve the host's specific and non-specific immunity, including cellular and humoral immunities, as well as promote recovery from myelosuppression in cyclophosphamide (CTX)-immunosuppressed mice. To our knowledge, this is the first report on chemical and immunoactivity study on polysaccharides from traditional Tibetan medicine compounds, which may provide a new idea for development of carbohydrate drugs from them.

Hormesis of methylmercury-human serum albumin conjugate on N9 microglia via ERK/MAPKs and STAT3 signaling pathways.

Methylmercury (MeHg+) is an extremely toxic organomercury cation that can induce severe neurological damage. Once it enters the body, methylmercury binds to amino acids or proteins containing free sulfhydryl groups. In particular, methylmercury is known to bind with human serum albumin (HSA) in human plasma; however, the effects of methylmercury-HSA conjugate (MeHg-HSA) on the central nervous system (CNS) are not fully understood. In the present study, we used the microglial cell line N9 as the target cells to evaluate the effect of MeHg-HSA on physiological function of the CNS preliminarily. The various factors in the cell culture were monitored by MTT assay, total lactate dehydrogenase assay, ELISA, qPCR, Western blot and flow cytometry techniques. The results showed that low-dose treatment with MeHg-HSA activated N9 cells, promoting cell proliferation and total cell number, enhancing NO and intracellular Ca2+ levels, and suppressing the release of TNFα and IL1ß without cytotoxic effects; while high-dose MeHg-HSA exhibited cytotoxic effects on N9 cells, including promoting cell death and increasing the secretion of TNFα and IL1ß. These results indicate that MeHg-HSA causes hormesis in microglia N9 cells. Furthermore, ERK/MAPKs and STAT3 signaling pathways related to the hormesis of MeHg-HSA on N9 cells. In addition, low dose of MeHg-HSA might be viewed as something very close to a lowest observed adverse effect level (LOAEL) for N9 cells. These findings will be useful for investigating the hormesis mechanism of MeHg+ and exploring the specific functions of MeHg-sulfhydryl conjugates on the central nervous system.

Viral etiology of acute respiratory infections in hospitalized children in Novosibirsk City, Russia (2013 - 2017).

BACKGROUND: Acute respiratory infections (ARIs) cause a considerable morbidity and mortality worldwide especially in children. However, there are few studies of the etiological structure of ARIs in Russia. In this work, we analyzed the etiology of ARIs in children (0-15 years old) admitted to Novosibirsk Children's Municipal Clinical Hospital in 2013-2017. METHODS: We tested nasal and throat swabs of 1560 children with upper or lower respiratory infection for main respiratory viruses (influenza viruses A and B, parainfluenza virus types 1-4, respiratory syncytial virus, metapneumovirus, four human coronaviruses, rhinovirus, adenovirus and bocavirus) using a RT-PCR Kit. RESULTS: We detected 1128 (72.3%) samples were positive for at least one virus. The most frequently detected pathogens were respiratory syncytial virus (358/1560, 23.0%), influenza virus (344/1560, 22.1%), and rhinovirus (235/1560, 15.1%). Viral co-infections were found in 163 out of the 1128 (14.5%) positive samples. We detected significant decrease of the respiratory syncytial virus-infection incidence in children with increasing age, while the reverse relationship was observed for influenza viruses. CONCLUSIONS: We evaluated the distribution of respiratory viruses in children with ARIs and showed the prevalence of respiratory syncytial virus and influenza virus in the etiological structure of infections. This study is important for the improvement and optimization of diagnostic tactics, control and prevention of the respiratory viral infections.

Hormesis of mercuric chloride-human serum albumin adduct on N9 microglial cells via the ERK/MAPKs and JAK/STAT3 signaling pathways.

Mercury chloride (HgCl2), a neurotoxicant that cannot penetrate the blood-brain barrier (BBB). Although when the BBB are got damaged by neurodegenerative disorders, the absorbed HgCl2, mainly in form of Hg (II)-serum albumin adduct (Hg-HSA) in human plasma, can penetrate BBB and affect central nervous system (CNS) cells. Current study planned to evaluate the effect of Hg-HSA on the physiological function of N9 microglial cells. At low dosage (15 ng/mL) of Hg-HAS, the observed outcomes was: promoted cell propagation, Nitric Oxide (NO) and intracellular Ca2+ levels enhancement, suppressed the release of TNF-α and IL-1ß and inhibited cell proliferation. At high dosage (15 µg/mL) we observed decline in NO and intracellular Ca2+ levels, and increment in the release of TNF-α and IL-1ß. These biphasic effects are similar to hormesis, and the hormesis, in this case, was executed through ERK/MAPKs and JAK/STAT3 signaling pathways. Study of quantum chemistry revealed that Hg2+ could form stable coordination structures in both Asp249 and Cys34 sites of HSA. Although five-coordination structure in Asp249 site is more stable than four-coordination structure in Cys34 site but four-coordination structure is formed easily in-vivo in consideration of binding-site position in spatial structure of HSA.

An antioxidant α-glucan from Cladina rangiferina (L.) Nyl. and its protective effect on alveolar epithelial cells from Pb2+-induced oxidative damage.

Air pollution is a serious global health problem nowadays. So, it is an emergency to pay sufficient attention to treat and prevent the diseases caused by air pollution, especially respiratory disease and lung damage. Cladina rangiferina (L.) Nyl. is an edible lichen that has been used in medicinal diets to treat respiratory and other diseases for over 500 years. In this study, a water-soluble polysaccharide, CRWP-P, was obtained from C. rangiferina by hot-water extraction, freeze-thawing separation, and Fehling reagent purification. Structural analysis showed that CRWP-P is a linear α-(1 → 3),(1 → 4)-d-glucan without branches. Its Mw was determined to be 1.05 × 105 Da. Its (1,3)-α-d-glucopyranosyl: (1,4)-α-d-glucopyranosyl ratio is approximately 1:2. Antioxidant activity assay showed that C. rangiferina polysaccharides, especially CRWP-P, had appreciable DPPH radical-scavenging activity and reducing power. Notably, they could effectively decrease cell breakdown and ROS generation, inhibit lipid peroxidation, increase key antioxidase activity, and promote glutathione redox cycling in Pb2+-oxidative injured A549 alveolar epithelium cells. Overall, the results of this study indicated that C. rangiferina polysaccharides, especially CRWP-P, have the potential to be natural antioxidants for the treatment of lung oxidative damage induced by lead of air pollutants.

pH-Dependent Effects of L-Cysteine on Mercury Dissolution of α-HgS and ß-HgS.

Mercury sulfide is an insoluble inorganic mercury compound, and it is the main chemical form in traditional oral mercury-containing medicines. Hg2+ has a high affinity for thiols, and small molecule thiols in the gastrointestinal tract may promote mercury dissolution of mercury sulfide by binding to Hg2+. L-cysteine is the only amino acid that possesses a reducing sulfhydryl group (-SH), out of the 20 amino acids. This study investigates the effect of L-cysteine on mercury dissolution of mercury sulfide at pHs ranging from 1.2 to 7.2. The results showed that L-cysteine had different pH-dependent effects on the mercury dissolution of α-HgS and ß-HgS. For α-HgS, the dissolved mercury concentration increased from 5.47 ± 0.97 ng/mL to 12.49 ± 0.54 ng/mL when the pH rose from 1.2 to 4.2, and decreased to 3.37 ± 0.70 ng/mL at pH 6.0 and then increased to 9.36 ± 0.79 ng/mL at pH 7.2. For ß-HgS, the dissolved mercury concentration increased from 151.09 ± 2.25 ng/mL to 2346.71 ± 62.62 ng/mL when the pH increased from 1.2 to 7.2. In conclusion, L-Cys was distinctly enhanced upon mercury dissolution of α-HgS and ß-HgS with increasing pH. These results may contribute to our understanding of the mercury absorption mechanism of traditional oral mercury-containing medicines.

A homogalacturonan from Hippophae rhamnoides L. Berries enhance immunomodulatory activity through TLR4/MyD88 pathway mediated activation of macrophages.

Our previous study isolated a natural high-methoxyl homogalacturonan (HRWP-A) from Hippophae rhamnoides and showed antitumor activity in vivo. In this study, the immunomodulatory activity and mechanisms of action of HRWP-A were further investigated. Results showed that HRWP-A could recover the body condition and activated macrophage in Cyclophosphamide (CTX)-induced immunosuppressed mice. Further, we investigated the possible mechanism underlying the effects of HRWP-A on mouse peritoneal macrophages. qPCR and western blot revealed that HRWP-A upregulated the expression of TLR4 mRNA in vitro. This process was accompanied by a clear increase in MyD88 expression and p-IκB-α, but these effects were largely abrogated by pretreatment with anti-TLR4 antibodies. The effects of HRWP-A on macrophage NO, IL-1ß and IL-6 production were also inhibited by anti-TLR4 antibodies and were greatly influenced by the NF-κB inhibitor PDTC. Moreover, HRWP-A failed to induce the production of NO, IL-1ß and IL-6 in peritoneal macrophages prepared from C3H/HeJ mice, which have a point mutation in the Tlr4 gene, suggesting the involvement of the TLR4 molecule in HRWP-A-mediated macrophage activation. These results may have important implications for our understanding of the structure-activity relationship of immunopotentiating polysaccharides from medicinal herbs.

Anticancer and immunostimulating activities of a novel homogalacturonan from Hippophae rhamnoides L. berry.

Our previous study isolated an anti-fatigue polysaccharide (HRWP) from the Hippophae rhamnoides berry. In this study, using ion-exchange chromatography and gel filtration chromatography in turn, a water-soluble homogenous polysaccharide HRWP-A was isolated from HRWP. Structural analysis determined that HRWP-A was a polysaccharide with repeating units of (1→4)-ß-d-galactopyranosyluronic residues, of which 85.16% were esterified with methyl groups. An antitumor activity assay showed that HRWP-A could significantly inhibit the Lewis lung carcinoma (LLC) growth in tumor-bearing mice. Further experiments suggested that the antitumor effect of HRWP-A might be mediated through immunostimulating activity, as it enhances the lymphocyte proliferation, augments the macrophage activities, as well as promoting NK cell activity and CTL cytotoxicity in tumor-bearing mice. To our knowledge, this is the first report on a natural antitumor high-methoxyl homogalacturonan pectin from the H. rhamnoides berry-a compound that acts as a potential immunostimulant and anticancer adjuvant.

Structural elucidation and antioxidant activity of a water-soluble polysaccharide from the fruit bodies of Bulgaria inquinans (Fries).

The non-lichenized ascomycete Bulgaria inquinans (Fries), growing in the Changbai Mountain of China, has been used as medicinal diet for many years. In a previous study, we have reported that a heteropolysaccharide BIWS-4b from the fruit bodies of B. inquinans (Fries) exhibited markedly antimalarial and immunostimulating activities. In this paper, the structural features and antioxidant activity of BIWS-4b were investigated. The results showed that BIWS-4b contains an α-(1→2), (1→6)-mannan core to which the glucogalactan chains are attached. The glucogalactan chains were composed of (1→6)-, (1→5)- and (1→5,6)-linked ß-Galf, (1→4)-linked and non-reducing terminal ß-Glcp units, and might be attached to the mannan core at the O-2 positions of α-Manp units. The antioxidant assays showed that BIWS-4b exhibited good activities, including free radicals scavenging effects, ferrous ion-chelating ability and reducing power. Thus, BIWS-4b could be used as a natural antioxidant agent for food and pharmaceutical industries.

The photocytotoxicity of different lights on mammalian cells in interior lighting system.

In the present paper, two light sources commonly used in interior lighting system: incandescent light and light emitting diode (LED) were chosen to evaluate their influences on three kinds of mammalian cells, together with UVA and UVB, and the mechanism of the photocytotoxicity was investigated in terms of intracellular ROS production, lipid peroxidation, SOD activity and GSH level assays. The results showed that LED and incandescent light both had some photocytotoxicities. In the interior lighting condition (100lx-250lx), the cytotoxicities of LED and incandescent lamp on RF/6A cells (rhesus retinal pigment epithelium cell line) were stronger than that on two fibroblast cell lines, while the cytotoxicity of UVA and UVB on HS68 cells (fibroblast cell line) was highest in the tests. The mechanism analysis revealed that the photocytotoxicities of LED and incandescent lamp were both caused by cell lipid peroxidation. LED and incandescent light could promote the production of ROS, raise lipid peroxidation level and lower the activity of the antioxidant key enzymes in mammalian cells, and finally cause a number of cells death. However, the negative function of LED was significantly smaller than incandescent light and ultraviolet in daily interior lighting condition. And the significantly lower photocytotoxicity of LED might be due to the less existence of ultraviolet. Therefore, LED is an efficient and relative safe light source in interior lighting system, which should be widely used instead of traditional light source.

Preparation of a novel glucuronomannan from Auricularia auricala and its immunological activity.

A glucuronomannan (AA-4-H, Mw around 4 KDa) was prepared from the fruit bodies of Auricularia auricala by extraction with hot water, deproteination by Sevag reagent, stepwise precipitation with ethanol and partial acid hydrolysis. Monosaccharides analysis revealed that AA-4-H consisted of 91% mannose (Man) and 9% glucuronic acid (GlcA). FT-IR, NMR and methylation analyses indicated that AA-4-H is a branched glucuronomannan. Its main chains are composed of 1, 3-linked alpha-Manp, side chains are single a-Manp or alpha-GlcA residues attached to the O-2 and O-6 of Man residues of the main chains. Bioassay indicated that AA-4-H remarkably enhanced B lymphocyte proliferation and increased the production of nitric oxide of macrophages in vitro. Thus, glucuronomannan AA-4-H could be explored as a potential immunostimulation agent.

Antioxidant and immunological activities of water-soluble polysaccharides from Aconitum kusnezoffii Reichb.

Aconitum kusnezoffii Reichb., one of the earliest recorded toxic species of genus Aconitum, has been used as traditional Chinese medicine and medicinal diet over the last 2500 years to treat heart failure congestion, neuralgia, rheumatism and gout, etc. In the present paper, four water-soluble polysaccharide fractions isolated from the tubers of A. kusnezoffii Reichb. were studied the antioxidant and immunological activities for the first time. In vitro antioxidant assays indicated that fraction WKCP-A had noticeable scavenging activities on DPPH radical, hydroxyl radical, superoxide anion, H(2)O(2) and self-oxidation of 1,2,3-phentriol, ferrous ion-chelating ability and reducing power. Moreover, the in vivo immunological assay exhibited that fractions WKCP-A and WKHP could more significantly enhance splenic lymphocyte proliferation and macrophage phagocytosis than other fractions. Therefore, the water-soluble polysaccharides from A. kusnezoffii Reichb., especially WKCP-A, have the potential to be explored as novel natural antioxidants and immunostimulating agents for using in functional foods or medicine.

The antitumor and immunostimulating activities of water soluble polysaccharides from Radix Aconiti, Radix Aconiti Lateralis and Radix Aconiti Kusnezoffii.

In the present study, the water-soluble polysaccharides of Radix Aconiti, Radix Aconiti Lateralis and Radix Aconiti Kusnezoffii, were extracted and fractionated into four fractions of each material. The FT-IR and chemical analyses indicated the water-soluble polysaccharides of the three materials were all mainly composed of starch, non-starch type alpha-D-glucans and pectic polysaccharides with different molecular weight distributions and monosaccharide composition ratios. The antitumor assay showed that all the non-starch type polysaccharide fractions had good antitumor activities, and the tumor growth inhibition ratios were 37.24-70.42%. Specifically the inhibition ratios of pectic polysaccharides were over 60%. Moreover, the immunological tests using the Cyclophosphamide (Cy) induced immunosuppressive mice, including phagocytosis of macrophage, NK cell activity, concanavalin A (ConA)-induced T-cell proliferation, lipopolysaccharide (LPS)-induced B-cell proliferation, quantitative haemolysis of sheep red blood cells (SRBC) and dinitro-fluorobenzene (DNFB)-induced delayed-type hypersensitivity (DTH) response assays, exhibited that all the non-starch type polysaccharides, especially the pectic polysaccharide fractions, not only had remarkable immunostimulating activities including nonspecific immunity, cellular immunity and humoral immunity, but also could restore the antitumor drug-suppressed immune function. Therefore, the polysaccharides from Aconitum species might be conveniently exploited to be good immune stimulating modifiers and had the potential to apply in the tumor therapy.