Selenochemical modification of low molecular weight polysaccharides from Grifola frondosa and the mechanism of their inhibitory effects on gastric cancer cells.

An important micronutrient involved in immune response and antitumor is selenium. LMW-GFP, a polysaccharide extracted from Grifola frondosa seed bodies, has a relatively weak antitumor effect on BGC-823 and MFC cells in vitro, whereas selenium binding to LMW-GFP can significantly increase the in vitro antitumor activity of LMW-GFP. In this study, Se-LMW-GFP was prepared by the HNO3-Na2SeO3 method, and the structures of LMW-GFP and Se-LMW-GFP were characterized by UV-visible spectroscopy of absorption, FTIR spectroscopy, and electron scanning microscopy, and these structural analyses showed that selenium was successfully complexed to LMW-GFP. The selenium content of Se-LMW-GFP was measured to be 2.08 % ± 0.08 % by ICP-MS. The anti-tumor activity of LMW-GFP before and after selenium modification was compared by cellular experiments, and the findings indicated that the anti-tumor activity of Se-LMW-GFP was considerably improved over that of LMW-GFP, and inhibited the proliferation of BGC-823 cells and MFC cells through a combination of the Fas/FasL-mediated exogenous death receptor pathway as well as the endogenous mitochondrial pathway. Our results suggest that Se-LMW-GFP not only has great potential for natural health food and anti-gastric cancer drug development but is also a good selenium supplement.

Structural characteristics of a low molecular weight velvet antler protein and the anti-tumor activity on S180 tumor-bearing mice.

Velvet antler is a traditional Chinese medicine with various pharmacological values, which is an important raw material for traditional Chinese medicinal wine. Nevertheless, the chemical compositions and bioactivities of velvet antler residue used for making medicinal wine are rarely reported, leading to a waste of resources. In this study, a velvet antler protein (VA-pro) was extracted from velvet antler residue by simulating the gastrointestinal digestion, and its composition, structural characteristics and in vivo anti-tumor activities were determined and investigated. VA-pro possessed high purity with a relatively low molecular weight as 22.589 kDa under HPLC, one- and two-dimensional electrophoresis, and it contained high contents of Pro, Gly, Glu and Ala. Besides, the secondary structure of VA-pro was dominated by ß-turn and ß-sheet, and VA-pro possessed similar protein sequence, isoelectric point and amino acid compositions to hypothetical protein G4228_020061. The in vivo results substantiated that VA-pro could improve the body weights and immune organ indices, increase the expressions of sera cytokines and regulate the distributions of T and B lymphocytes subsets in peripheral blood of S180 tumor-bearing mice. Furthermore, VA-pro could effectively inhibit solid S180 tumors growth by inducing S phase cell cycle arrest mediated through mitochondria. To summarize, our study provided theoretical support that VA-pro had the potential to be used as an immunopotentiator in immunocompromised or cancer-bearing hosts.

Flow-induced "waltzing" red blood cells: Microstructural reorganization and the corresponding rheological response.

We investigate flow-induced structural organization in a dilute suspension of tumbling red blood cells (RBCs) under confined shear flow. For small Reynolds (Re = 0.1) and capillary numbers (Ca), with fully coupled hydrodynamic interaction (HI) and without interparticle adhesion, we find that HI between the biconcave discoid particles prompts the formation of layered RBC chains and synchronized rotating RBC pairs, referred here as "waltzing doublets." As the volume fraction ϕ increases, more waltzing doublets appear in RBC files. Stronger shear stress disrupts structural arrangements at higher Ca. We find that the flow-induced organization of waltzing doublets changes how the suspension viscosity varies with ϕ qualitatively. The intrinsic viscosity is particularly sensitive to microstructural rearrangement, increasing (decreasing) with ϕ at low (high) Ca that correlates with the change in the fraction of doublets. We verified flow-induced collective motion with comparison to two-cell simulations in which the cell volume fraction is controlled by varying the domain volume.

Structural Characterization of an Alcohol-Soluble Polysaccharide from Bletilla striata and Antitumor Activities in Vivo and in Vitro.

In general, Bletilla striata polysaccharides were mostly water-soluble. However, the structural property, immunomodulatory effects and antitumor activities of alcohol-soluble Bletilla striata polysaccharide were rarely reported. In this study, an alcohol-soluble Bletilla striata polysaccharide was firstly extracted, investigated the structural property and evaluated the antitumor activity in vivo and in vitro. Results showed that BSAP was a low molecular weight polysaccharide (2.29×104  Da) and consisted of glucose, xylose and mannose (molar ratio: 2.39 : 1.00 : 0.21). Animal experiments results suggested that BSAP could effectively inhibit the expansion of H22 solid tumors, protect thymus and spleen, improve macrophages, lymphocytes and NK cells activities and enhance lymphocyte subsets proportion, presenting a better immunological enhancement effect in vivo. Additionally, the results of cell experiments showed that BSAP had obvious antitumor effect in vitro, including inhibiting the proliferation of H22 cells and inducing the apoptosis of tumor cells. These results would provide theoretical basis and new ideas for the further development and utilization of BSAP in the biomedical field.

Preparation, Antioxidant and Immunoregulatory Activities of a Macromolecular Glycoprotein from Salvia miltiorrhiza.

Salvia miltiorrhiza has exhibited various bioactive functions due to the existence of polysaccharides, hydrophilic phenolic acids, diterpenoid quinones, and essential oils. However, little research has reported the glycoprotein preparation and corresponding bioactivities. In this study, the water-soluble glycoprotein from S. miltiorrhiza roots was firstly isolated with the extraction process optimized by response surface methodology, and then, the preliminary structural properties, and the antioxidant and immunoregulatory activities were investigated. Results showed that the extraction conditions for higher extraction yields were identified as follows: ultrasonic power of 220 W, ultrasonic time of 2.0 h, extraction temperature of 60 °C, liquid/solid ratio of 20 mL/g, and the glycoprotein yields of 1.63 ± 0.04%. Structural analysis showed that the glycoprotein comprised protein and polysaccharide (contents of 76.96% and 20.62%, respectively), with an average molecular weight of 1.55 × 105 Da. Besides, bioactivities analysis showed that the glycoprotein presented strong scavenging effects on multiple free radicals, and effectively enhanced the antioxidant enzyme activities and immunological indicators in cyclophosphamide-induced immunocompromised mice dose-dependently. These data demonstrated that S. miltiorrhiza glycoprotein presented the potential to be a novel edible functional compound, and could be practically applied in the food industry.

Ultrasonic-Assisted Extraction of Codonopsis pilosula Glucofructan: Optimization, Structure, and Immunoregulatory Activity.

In recent years, multiple edible polysaccharides from Codonopsis pilosula were mainly isolated with high average molecular weights and exhibited various bioactivities, but it was proven that low-molecular-weight polysaccharides could exert stronger activities due to the superior water solubility and permeability. In the present study, the water-soluble polysaccharide C. pilosula with low molecular weight was isolated under ultrasonic assistance at 30 °C, the extraction process was optimized via response surface method (RSM), and the structure and immunoregulatory activity were further investigated. The maximum yield (4.86%) for crude polysaccharides (cCPPs) was obtained under following parameters: ultrasonic power of 370 W, liquid/material ratio of 33 mL/g, ultrasonic time of 81 min. Subsequently, the cCPPs were further purified through dialysis and Sephadex G-25 column to acquire purified polysaccharide (CPPs). Structural analysis indicated that CPPs was a glucofructan (average molecular weight of 4.23 × 103 Da) with (2→1)-ß-D-Fruf and (1→)-α-D-Glcp as the backbone branched by (2→6)-ß-D-Fruf. Additionally, CPPs could enhance immunoregulatory function by stimulating NO production and cytokine (IL-6 and TNF-α) secretion of RAW264.7 macrophages dose-dependently, which presented no cytotoxic effects. These data suggest that CPPs have the potential to be used as a nutritional dietary compound and natural immunostimulant supplement in the food industry.

Structural characterization of a low molecular weight Bletilla striata polysaccharide and antitumor activity on H22 tumor-bearing mice.

In this study, a novel low molecular weight polysaccharide (named LMW-BSP) was extracted from Bletilla striata at 4 °C. The results of structural characteristics analysis showed that LMW-BSP was a 23 kDa neutral polysaccharide contained glucose and mannose at a molar ratio of 1.00:1.26. Structural investigations of the periodate oxidation studies, Smith-degradation as well as methylation were performed, and combined with 1D and 2D NMR spectroscopy, the main chain residues sequence of LMW-BSP was concluded to be: α-D-Manp-(1 â†’ 3)-ß-D-Manp-(1 â†’ [4)-ß-D-Glcp-(1]2 â†’ 4)-ß-D-Manp-(1 â†’ 3)-ß-D-Manp-(1→. Moreover, the antitumor activity of LMW-BSP was evaluated in H22 tumor-bearing mice. And the results suggested that LMW-BSP could effectively improve immune cells activities and lymphocytes subsets proportions dose-dependently in tumor-bearing mice, leading to the apoptosis of H22 cells via G1 phase arrested. LMW-BSP inhibited tumor growth and exhibited antitumor effects in vivo. And it supported considering the novel polysaccharide as a potential drug component in hepatocellular carcinoma treatment.

Structural characterization of a water-soluble polysaccharide from Angelica dahurica and its antitumor activity in H22 tumor-bearing mice.

A novel Angelica dahurica polysaccharide (ADP) with Mw of 6.09 × 103 Da was isolated. The contents of total sugar and uronic acid in ADP were 91.04% and 12.69%. The structure characteristics indicated that ADP was an acidic polysaccharide consisting of rhamnose, arabinose, galactose, glucose, mannose, glucuronic acid and galacturonic acid (0.09: 0.61: 1.88: 1: 0.14: 0.63: 0.03). Moreover, there were →3)-Manp-(1→, →4, 6)-Galp-(1→, →4)-Galp-(1→, →3)-Glcp-(1→, →5)-Araf-(1→, →2)-Galp-(1→ in ADP with relative molar ratios of 0.32:0.57:0.29:0.95:0.71:0.26. In vivo experiments suggested that ADP significantly inhibited the tumor growth of mice, increased the activities of spleen lymphocytes and natural killer (NK) cells, improved the cytokine level (IL-2 and TNF-α) and the proportions of lymphocyte subsets in the peripheral blood. The tumor cell progression was arrested in the G1 phase, and the apoptosis rate of tumor cells were 7.54% and 19.32% at the dose of 100 and 200 mg/kg, which was consistent with the results of pathological observation. In summary, the study might provide a theoretical basis for the application on functional foods containing Angelica dahurica polysaccharides.

Antitumor and immunoregulatory activities of a novel polysaccharide from Astragalus membranaceus on S180 tumor-bearing mice.

The Astragalus membranaceus polysaccharide (APS4) with direct cytotoxicity on various cancer cells has been prepared in our previous study, while the underlying therapeutic role of APS4 on solid tumors in vivo hasn't been investigated yet. Therefore, in this paper, the lymphocytes-mediated antitumor and immunoregulatory activities of APS4 were researched by establishing S180 tumor-bearing mice model. Flow cytometry analysis revealed that APS4 could effectively regulate the percentages of CD3+, CD4+, CD8+ T cells and CD19+ B cells in thymus, peripheral blood and spleen of S180 tumor-bearing mice, dose-dependently. H&E staining and cell cycle determination of solid tumors manifested that APS4 treatment could significantly inhibit the growth of solid tumors by inducing cells apoptosis. Furthermore, two-dimensional electrophoresis and western blot analysis further demonstrated that APS4 could activate antitumor-related immune cells and promote anaerobic metabolism of tumor microenvironment, thereby causing the apoptosis of S180 tumor cells. These data implicated that APS4 could be used as a potential dietary supplement for immune enhancement.

The immunosuppressive effects of low molecular weight chitosan on thymopentin-activated mice bearing H22 solid tumors.

In the present study, the low molecular weight of chitosan (CS) was prepared and its activity on thymopentin-activated mice bearing H22 solid tumors was further researched. The purity and molecular weight of CS were determined by UV and HPGPC spectra, and its immunosuppressive effects on H22 tumor-bearing mice were evaluated through determination on immune organs, cells and cytokines. Results showed that CS contained little impurities with the average molecular weight of 1.20 × 104 Da. The in vivo antitumor experiments demonstrated that CS facilitated to destroy immune organs (thymuses and spleens), suppress immune cells (lymphocytes, macrophages and NK cells) activities and reduce immune-related cytokines (TNF-α, IFN-γ, IL-2 and IL-4) expressions of H22 tumor-bearing mice even with simultaneous TP5 stimulation. Our data suggested that CS could not be applied to improve immune response in cancer-bearing patients, but might be employed for treatments on autoimmune diseases or organ transplant patients.

A novel acid polysaccharide from Boletus edulis: extraction, characteristics and antitumor activities in vitro.

A novel cold-water-soluble polysaccharide (BEP), with a molecular weight of 6.0 × 106 Da, was isolated from Boletus edulis. BEP consists of galactose, glucose, xylose, mannose, glucuronic, and galacturonic acid in a ratio of 0.34:0.28:0.28:2.57:1.00:0.44. The IR results showed that BEP was an acid polysaccharide, containing α-type and ß-type glucoside bonds. MTT assay showed BEP could inhibit cell proliferation significantly. Morphological observation demonstrated that BEP-treated MDA-MB-231 and Ca761 cells exhibited typical apoptotic morphological features. Flow cytometry analysis revealed that BEP caused mitochondrial membrane potential collapse. Annexin V-FITC/PI staining indicated that BEP induced apoptosis of MDA-MB-231 and Ca761 cells through cell block in S phase and G0/G1 phase, respectively. Western blot results showed that BEP could increase the Bax/Bcl-2 ratios, promote the release of cytochrome C, and activate the expression of caspase-3 and caspase-9 in MDA-MB-231 and Ca761 cells. In conclusion, our results demonstrated that BEP could inhibit the proliferation of breast cancer cells and induce apoptosis through mitochondrial pathways.

A Novel Optimization of Water-Soluble Compound Polysaccharides from Chinese Herbal Medicines by Quantitative Theory and Study on Its Characterization and Antioxidant Activities.

The present study optimized the extraction characterization and antioxidant activities of water-soluble compound polysaccharides (CPs) from hawthorn, lotus leaf, Fagopyrum tataricum, semen cassiae, Lycium barbarum, and Poria cocos Chinese herbal medicines that have mass ratios of 4 : 2 : 2 : 1.5 : 1 : 1. The CPs yield equation was predicted using quantitative theory, to which a maximum CPs yield of 7.18±0.24 % under the following optimal extraction conditions: a water-to-raw material ratio of 30 mL/g, an extraction temperature of 65 °C, an extraction time of 45 min, and extraction mode ultrasonic-assistant extraction. CPs were consisted of Ara, Gal, Glc, Xyl, Man, GalA and GlcA in a molar ratio of 3.1 : 2.6 : 50.6 : 1.7 : 20.4 : 17.2 : 4.2. The HPGPC profiles and FT-IR spectra implied that CPs were heterogeneous acidic polysaccharides and possessed the ß-d-pyranose configuration. Congo red test, CD spectrum and SEM revealed that CPs with three helix conformation showed a flocculent, granulous or sheet-like appearance. Furthermore, the relationships between antioxidant activity and concentration of CPs displayed significant positive correlation, and the scavenging abilities for DPPH, hydroxyl radical, ABTS, superoxide-anion radical and reducing power of CPs were 93.56±2.51 %, 84.03±1.69 %, 83.29±1.93 %, 37.49±1.93 % and 0.467±0.006 at a concentration of 4.0 mg/mL. Therefore, CPs could be applied as a potential natural antioxidant in pharmaceutical or functional food fields.

The preparation of a cold-water soluble polysaccharide from Grifola frondosa and its inhibitory effects on MKN-45 cells.

In this study, a novel water soluble polysaccharide (named GFP-4) was extracted from Grifola frondosa at 4 oC, and its preliminary structure and inhibitory effects on human gastric carcinoma MKN-45 cells through the Fas/FasL death receptor apoptosis pathway were investigated. High-performance gel permeation chromatography (HPGPC), fourier-transform infrared spectroscopy (FT-IR), and ion chromatography (IC) results showed that GFP-4 was a 1.09 × 106 Da neutral hetero polysaccharide with pyranose rings, and α- and ß-type glycosidic linkages that contained galactose, glucose, and mannose at a molar ratio of 1.00:3.45:1.19. MTT results indicated that GFP-4 significantly inhibited the proliferation of MKN-45 cells in a concentration-dependent manner. The H&E staining and Hoechst 33342/PI double staining results showed that GFP-4-treated MKN-45 cells were subjected to underwent typical apoptotic morphologic changes such as nuclear pyknosis, chromatin condensation, and an increase of membrane permeability. Annexin V-FITC/PI double staining, cell cycle analysis, and western blot results revealed the GFP-4 induced MKN-45 cells apoptosis through the Fas/FasL-mediated death receptor pathway with cells arrested at the G0/G1 phase. These data indicate that GFP-4 is a promising candidate for treating gastric cancer and provide a theoretical basis for the future development and utilization of G. frondosa clinically.

The structural characteristics of an acid-soluble polysaccharide from Grifola frondosa and its antitumor effects on H22-bearing mice.

The edible mushroom G. frondosa has been used as a kind of functional food for the prevention and therapy of various diseases in Asian countries. In the present work, a novel acid-soluble polysaccharide (GFAP) was successfully isolated from G. frondosa under room temperature and hydrochloric acid solution treatment. Results of chemical composition analysis, UV and HPGPC spectra showed that GFAP mainly contained 94.28% of carbohydrate with the average molecular weight of about 644.9 kDa. GC, FT-IR, NMR and methylation analysis further indicated that GFAP was a neutral sugar mainly composed of (1 → 3)-ß-D-Glcp and (1 → 3)-α-D-Manp. The in vivo antitumor experiments demonstrated that GFAP could effectively protect thymuses and spleens of tumor-bearing mice and inhibit the growth of H22 solid tumors with the inhibitory rate of 36.72%. Besides, GFAP could significantly improve the activities of NK cells, macrophages, CD19+ B cells and CD4+ T cells, leading to the apoptosis of H22 cells via G0/G1 phase arrested. Our data demonstrated that GFAP holds great application prospect to be a safe and effective antitumor adjuvant in the future.

The caspases-dependent apoptosis of hepatoma cells induced by an acid-soluble polysaccharide from Grifola frondosa.

The edible mushroom Grifola frondosa has been used as a functional food for diseases prevention and therapy in Asian countries. In the present work, an acid-soluble polysaccharide (GFAP) was prepared from Grifola frondosa under room temperature and hydrochloric acid solution treatment, and its inhibitory effects on H22 and HepG2 cells were investigated. Results of MTT indicated that GFAP could effectively suppress proliferations of HCC cells, dose-dependently. Microscopic observation results demonstrated that GFAP-treated HCC cells showed apoptotic characteristics like membrane blebbing, chromatin condensed, nucleus pycnosis and fragmentation. Annexin V-FITC/PI staining and cell cycle distribution results showed that GFAP could induce the apoptosis of H22 and HepG2 cells via arresting them in G1 and S phases respectively. Rh123, JC-1 staining and western blotting results suggested that GFAP could significantly increase the permeability of mitochondrial membrane of HCC cells, and upregulated the expressions of Bax, cytochrome c, cleaved-caspase-3 and cleaved-caspase-9, which indicated that GFAP could trigger apoptosis of HCC cells through mitochondria apoptotic pathway in a caspases-dependent pattern. Our data demonstrated that GFAP holds great application prospect as a safe and effective antitumor drug for hepatocellular carcinoma therapy.

Seleno-Chitosan induces apoptosis of lung cancer cell line SPC-A-1 via Fas/FasL pathway.

In this study, human lung cancer SPC-A-1 cells were cultured with Seleno-Chitosan to study the mechanism of apoptosis. CCK-8 results showed that with the increase of the concentration of Seleno-Chitosan and the prolongation of culture time, the inhibition on the proliferation of SPC-A-1 cells gradually increased, and the morphology of SPC-A-1 cells changed obviously. Flow cytometry result suggested that the concentration of calcium ion, level of reactive oxygen species and mitochondrial membrane potential increased. Western blot showed that Seleno-Chitosan induced apoptosis via the mitochondrial pathway in SPC-A-1. Eventually, we found 15 proteins that were expressed abnormally by Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), which were all related to Fas/FasL pathway of cells.

Relationship between heat treatment on structural properties and antitumor activity of the cold-water soluble polysaccharides from Grifola frondosa.

Grifola frondosa is a basidiomycete fungus with potential biomedical applications owing to the presence of bioactive polysaccharides. The activities of polysaccharides are influenced by many factors, particularly temperature; however, the optimal temperature and conditions for preparation of polysaccharides from this organism have not yet been determined. Therefore, in this study, cold-water soluble polysaccharides from Grifola frondosa were extracted at 4 °C (GFP-4) and purified. GFP-4-30, GFP-4-60 and GFP-4-90 were obtained from GFP-4 after treatment at 30 °C, 60 °C, or 90 °C, respectively, for 6 h. MTT results showed that GFP-4 had the highest inhibitory effects on the proliferation of SPC-A-1 cells in vitro. High-performance gel permeation chromatography results demonstrated that the molecular weight of GFP-4 was 1.05 × 106 Da and that GFP-4-30, GFP-4-60, and GFP-4-90 showed different levels of degradation and generated small molecule sugars. Fourier transform infrared spectroscopy, gas chromatography, and nuclear magnetic resonance results indicated that GFPs mainly consisted of α-D-Galp, α-D-Manp and α-D-Glcp. Periodate oxidation, Smith degradation, and methylation results showed that the backbones of the molecules consisted of 1,3-linked-Galp. After heat treatment, percentages of (1 → 3,4) α-D-Galp in heat-treated polysaccharides were obviously decreased, indicating their lower branching degree, and resulting in weaker antitumor effects. Overall, our findings demonstrated changes in the structure-activity relationships of GFP-4 after heat treatment and provided a theoretical basis for the application of GFP-4 in the food and drug industries.

An alcohol-soluble polysaccharide from Atractylodes macrocephala Koidz induces apoptosis of Eca-109 cells.

In this study, polysaccharides from Atractylodes macrocephala Koidz (APA) which were soluble in alcohol were prepared, purified, analyzed the structure and investigated the antitumor activity in vitro cell experiment. Results of high-performance gel permeation chromatography (HPGPC), fourier-transform infrared spectroscopy (FT-IR), and gas chromatography (GC) showed that APA was a 2.1KDa neutral hetero polysaccharide composed of arabinose and glucose (molar ratio, 1.00:4.57) with pyranose rings and α-type and ß-type glycosidic linkages. Results by MTT experiments showed that the proliferation inhibition was 74.63% in Eca109 cells treated with 2 mg/mL dose of APA. Annexin V/PI assay, Hoechst 33,258 staining, cell cycle distribution, rhodamine 123 dye assay and western blot assay clarified that APA could accelerate the apoptosis of Eca109 cells by mitochondrial pathway and stocked cells at S phase. These data indicated that APA is a promising potential candidate for therapeutic treatment of esophageal cancer.

Seleno-ß-lactoglobulin (Se-ß-Lg) induces mitochondria-dependant apoptosis in HepG2 cells.

Selenium compounds have been widely investigated as novel anticancer agents due to high efficacy and selectivity against cancer cells in recent years. This study aimed to research the potential inhibitory effects of seleno-ß-lactoglobulin (Se-ß-Lg) on HepG2 cells in vitro. MTT results demonstrated that the synthetized Se-ß-Lg exhibited strong antitumor activity on HepG2 cells with few side effects on human normal cells (LO2) and relatively weaker cytotoxic effects compared to inorganic selenium (SeO2). Scanning electron microscope (SEM), hoechst 33342/PI double staining, annexin V-FITC/PI staining and cell cycle detection results showed that Se-ß-Lg could induce the apoptosis of HepG2 cells via arresting them in S and G2/M phases and lead to the obvious morphological changes (loss of adhesion, cell shrinkage, and membrane blebbing, membrane permeabilities and DNA fragmentation). Besides, JC-1 staining, western blotting (WB) and polymerase chain reaction (PCR) results showed that Se-ß-Lg could gradually destroy the mitochondrial membrane potential of HepG2 cells, and finally resulting in the mitochondria-dependant apoptosis via up-regulation of Bax, Cytochrome c, Caspase-3 and down-regulation of Bcl-2. Our data could provide a theoretical basis for practical application of Se-ß-Lg in food and drug industries.

Polysaccharide extracted from Atractylodes macrocephala Koidz (PAMK) induce apoptosis in transplanted H22 cells in mice.

Polysaccharide of Atractylodes macrocephala Koidz (PAMK) was extracted by alcohol sedimenting ranging from 60% to 90% and purified by ultrafiltration membrane. The high-performance gel permeation chromatography (HPGPC), Fourier-transform infrared spectroscopy (FT-IR), gas chromatography (GC) and Nuclear magnetic resonance (NMR) revealed that PAMK was a 4.1KDa neutral heteropolysaccharide composed of galactose, arabinose and glucose with α-configuration (molar ratio, 1: 1.5: 5). Results of determination of chemical components suggested that PAMK contained 96.47% of polysaccharide and little protein, nucleic acid and uronic acid. Antitumor experiments in vivo could be concluded that PAMK had a significantly cytotoxic and anti-tumor effect by blocking tumor cells in S phase. And not only that, PAMK could protect immune organ efficiently, comparing with cyclophosphamide. The research provided a potential antineoplastic drug component for tumor treatment.