以幼年期(10年以下)、挂果期(20年)和盛年期(40年以上)的银杏树根系土壤为研究对象,探讨银杏雌株树龄与土壤中细菌菌群结构的关系.运用Illumina MiSeq平台测序分析幼年期、挂果期以及盛年期银杏树根系土壤细菌的16S rRNA V3～V4区变异序列,对不同年龄阶段银杏树雌株根系土壤细菌的菌群多样性和结构进行研究.银杏树雌株根系土壤共获得612219条微生物的16S rRNA有效基因序列,在97％的相似水平下共得到OTU个数为23294.热图分析表明,各个年龄段银杏树雌株的土壤细菌群落差异较大,不同年龄段优势属不同.据9个土壤样品的多样性指数分析得出,在3组年龄段银杏雌株的土壤样本中,幼年期Shannon指数、Simpson指数、Chao1指数以及ACE指数都是最大,而挂果期皆最小,表明幼年期银杏雌株土壤的细菌菌群丰富度和多样性最高,挂果期银杏土壤的细菌菌群丰富度和多样性最低.变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)在土壤细菌群落中占绝对优势.挂果期和盛果期银杏雌株土壤中浮霉菌门(Planctomycetes)及变形菌门(Proteobacteria)和相对丰度普遍低于幼年期,挂果期银杏树的土壤与盛果期银杏树的土壤中的放线菌门(Actinobacteria)与绿弯菌门(Chloroflexi)的相对丰度明显高于幼年期银杏树的土壤.WS3在盛果期银杏土壤中的相对丰度最低.银杏树雌株树龄可对土壤细菌菌群的多样性及菌群结构产生影响.伴随着育种年限的增加,银杏雌株表层土壤中细菌多样性呈现先降低后增加的变化趋势,且银杏雌株树龄对其土壤细菌的多样性和结构有显著影响.

晒干的"南薯88"的茎叶,通过热水浸提,乙醇沉淀,DEAE-cellulose column柱层析分离纯化,得到"南薯88"多糖纯品（N88P-1）。通过红外光谱技术（IR）,高效凝胶渗透色谱（HPGPC）,高效液相色谱色谱（HPLC）和核磁共振谱（1 H NMR）等对"南薯88"多糖（N88P-1）的进行了结构鉴定;同时,对"南薯88"多糖（N88P-1）的抗氧化及免疫调控作用进行了初探。结果显示,"南薯88"多糖（N88P-1）的分子量为1 4328 D,红外光谱显示,在3 444.81 cm -1 出现O-H的特征伸缩振动峰,1 640.20 m -1 出现C=O的特征伸缩振动峰,~1H NMR谱显示δ4.93～δ4.86出现异头氢信号,高效液相色谱色谱显示N88P-1的单糖组成为1∶1的半乳糖和甘露糖。活性研究结果显示,一定范围内的N88P-1浓度可以清除ABTS + 自由基,IC 50 值为0.48 mg/mL。N88P-1在0.25～3 mg/mL的浓度范围内与DPPH - 自由基的清除能力呈正相关,IC 50 值为0.96 mg/mL。当N88P-1的浓度为3 mg/mL时,对ABTS + 和DPPH - 自由基的清除能力均达到最高值;免疫细胞增殖活性结果显示,N88P-1在20～80 mg/mL的浓度下,对B细胞和T细胞增殖效果为极显著（P＜0.01）;在10～40 mg/mL的浓度下,对巨噬细胞增殖效果为极显著（P＜0.01）,其中,N88P-1在40 mg/mL的浓度下对3种免疫细胞的增殖效果均达到最佳。

探究大熊猫核糖体蛋白基因RPL35A及其编码蛋白的特点,以进一步研究其编码蛋白的功能,为深入开展大熊猫分子生物学研究及大熊猫基因保护等积累科学资料;本研究以大熊猫骨骼肌为材料,采用RT-PCR和降落-PCR方法克隆RPL35A基因的cDNA基因组序列,IPTG法诱导基因在BL21细胞系中的表达;大熊猫RPL35A基因的cDNA序列为337 bp,编码110个氨基酸;该基因的基因组序列长1 275 bp,包含2个外显子和1个内含子;RPL35A基因在BL21细胞系中的表达产物与组氨酸标签蛋白融合形成18 kDa的融合蛋白;该基因的cDNA序列、基因组序列中的外显子片段及其编码的蛋白序列与已报道的人、牛、小家鼠、褐家鼠等物种的对应序列同源性很高,蛋白质理化性质和功能位点基本相同,这说明该基因及其编码蛋白在进化过程中非常保守,可用于分子进化方面的研究,在物种进化关系研究中具有一定的参考意义和研究价值;同时,该基因可在原核生物中表达出具有活性的产物,可进一步开展纯化和蛋白质功能的研究.

A new heteropolysaccharide was extracted and purified from the fruiting bodies of Cantharellus cibarius Fr. The Cantharellus cibarius Fr. polysaccharide (CC-1) had a molecular weight of 61,056 kDa and was mainly formed of the glucose and xylose at ratio of 5:1. Structure identification of CC-1 was analysed by a combined application of total hydrolysis, high performance liquid chromatography (HPLC), methylation analysis, gas chromatography-mass spectrometry (GC-MS), infrared (IR) spectra and nuclear magnetic resonance (NMR) spectroscopy. The experimental results showed that CC-1 had a backbone of 1,4-linked--D-glucose which branched at O-6 and the branches were mainly composed of 61)--D-xylopyranose residue. CC-1 exhibited significant in vitro antioxidant effect and proliferation effect of immune cells. The activity study showed CC-1 has ability to clear the ABTS(+) free radical and DPPH- free radical in a certain range of concentration. The proliferation activity of the immune cells showed that the proliferation effect on B cells was very significant (P<0.001) in the concentration of 0.625-80 mg/ml; and the effect of T cell proliferation was also very significant (P<0.001) in the concentration of 5-20 mg/ml. The result of this study introduced Cantharellus cibarius Fr. as a possible valuable source in exhibiting unique immunoregulatory and antioxidant properties.

The fundamental mechanisms underlying the preventional and therapeutic effects of polysaccharides from fungi, including the immunostimulatory, antiviral and antitumor effects, are considered to occur through the modulation and stimulation of the macrophage and complement system. LDG-A, a novel polysaccharide from Lactarius deliciosus (L. ex Fr.) Gray exhibits marked antitumor activities in vivo. However, the underlying molecular mechanism of the antitumor activities of LDG-A remains unclear. In the present study, cell cycle analysis was performed in macrophages and B cells, and the transcriptomes of macrophages in the control group and LDG-A group were sequenced using Illumina sequencing technology to analyze the differentially expressed genes (DEGs), and elucidate the molecular mechanisms underlying the immunomodulatory and antitumor activities of LDG-A. The cell cycle analysis results indicated that LDG-A was able to promote the proliferation of B cells by promoting cell cycle progression in S phase and G(2)/M phase and eliminating cell cycle arrest in G(0)/G(1), and promote the proliferation of macrophages by promoting cell cycle progression in G(0)/G(1) phase and eliminating cell cycle arrest in G(2)/M phase. Of the total number of genes (8,140), 77.00% were expressed [reads per kilobase per million reads (RPKM) 1] and 1,352 genes were highly expressed (RPKM >60) in the LDG-A group. Of 775 unigenes which were identified as DEGs, 469 were downregulated and 306 genes were upregulated. A protein chip method was also used to determine the cytokines secreted by macrophages. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis and GO enrichment analysis indicated that the Janus kinase/signal transducer and activator of transcription, mitogen-activated protein kinase, chemokine, vascular endothelial growth factor and transforming growth factor signaling pathways are markedly enriched for DEGs.

e14565 Background: H1 is a peptide with 15 AA which can bind to a cell membrane protein. H2 is a peptide with 12 AA which can bind to EGFR. The sequence of these two peptides are completely consistent with human proteins. It is used conduct experiment to assess the safety, pharmacokinetics (PK), pharmacodynamics (PD) and anti-tumor activity. Methods: H1 and H2 peptide are synthesized with Peptide Synthesizer. Some of H1 and H2 peptide are processed by using PEG. The experiment of Pharmacodynamics, pharmacokinetics, pharmacology, drug toxicity test and drug safety test are adopting conventional standards. An improved tomographic method was used for target protein combined with H1 peptide detection. H1 and H2 peptides are conjugated with FITC for Pharmacokinetics respectively. Results: H1 peptide can bind to a cell membrane protein of 47KD. H1 peptide can cause some tumor cell of cell line dead but not impact normal cell in vitro. The tumor growth inhibition rate of H1 peptide with the dose of 2 ug/kg was 69%, and rate of the Cyclophosphamide as positive contrast was 67% in vivo. The rate of H2 at the same dose was 53%. No pathological lesion of tissue and organs of experimental animal were observed by acute toxicity test and subchronic toxicity test at the dose of 400000 ug/kg of H1 or H2 peptide. The half-life period of H1 peptide in blood was 3.6 minutes, while the half-life period of H1 peptide with PEG processing was 156 minutes. Conclusions: H1 and H2 peptide have anti-tumor effect in vivo and in vitro. It is worth pointing out that the anti-tumor effect of H1 and H2 peptide is as strong as Cyclophosphamide but has no toxic side-effects.

To understand the effect of Lactarius deliciosus Gray polysaccharide (LDG-A) on the proliferation of specific immunocytes and tumor cell apoptosis,we tested and observed the proliferation and morphological changes of T lymphocytes and B lymphocytes in the presence of LDG-A,determined the number of T lymphocytes and B lymphocytes in each stage of the cell cycle,and also evaluated antibody secretion by B lymphocytes under the stimulation of LDG-A.In addition,the apoptosis and cytoskeleton of lung cancer cell A549 were evaluated.The results showed that the proliferation of B lymphocytes and T lymphocytes' proliferation was promoted LDG-A in a dose-response manner.LDG-A could also reduce the number of B lymphocytes at the G0/G1 phase and promote them to enter the G2/M phase.Meanwhile,LDG-A also promoted the secretion of IgM,IgD and IgE by B lymphocytes.In addition,LDG-A in the range of 12.5-50.0 μg/mL could induce the apoptosis of lung cancer cell A549 in a dose-dependent manner.Moreover,we observed fractured F-actin filaments in the cells as well as damaged protein fiber networks during apoptosis.In conclusion,LDG-A has a promoting effect on the proliferation of specific immunocytes,and accelerates the apoptosis of tumor cells in vitro.

A new heteropolysaccharide was isolated from the fruiting bodies of Lactikporus supharells (Fr.) Murr. The immune response of polysaccharides to the cells in vitro was further investigated. The heteropolysaccharide was named LSM-X, the structure of polysaccharide LSM-X was characterized by HPGPC, IR, HPLC, GC-MS, and NMR. The results of structural analysis showed that polysaccharide LSM-X consists mainly of beta-D-glucose, alpha-D-galactose and a-D-lyxose, which ratios were 4: 2: 1, respectively. The main chain was 1,6-linked beta-D-glucose, the 4-O had a side chain of two 1,6-linked alpha-D-galactose residues and a 1-linked alpha-D-lyxose residue. The results of immunomodulatory activity showed that polysaccharide LSM-X could promote B cells and T cells proliferation. When the concentration of LSM-X was 10 mu g/mL, the number of B cells and T cells were largest compare with blank control group. The B cell and T cell cycle were detected by flow cytometry. Under the stimulation of polysaccharide LSM-X, the number of B cells decreased significantly in G0/G1 phase, and the number of B cells increased significantly in G2/M phase, while the number of T cells increased significantly in G0/G1 phase, and the number of T cells decreased significantly in both S and G2/M phase. Thus, polysaccharide LSM-X could promote B cell and T cell proliferation by regulating the cycle of B cells and T cells, respectively. In summary, this study identified the structure of LSM-X and explored the proliferation of immune cells. It also provided scientific basis for future studies on biological activity.

Acute lung injury (ALI) is an early pathophysiologic change in acute respiratory distress syndrome and its management can be challenging. Omalizumab (Xolair™) is a recombinant DNA-derived, humanized antibody. OMZ-SPT is a polypeptide on the heavy chain of omalizumab monoclonal antibody. Here, we found that intramuscular administration of OMZ-SPT significantly improved survival and attenuated lung inflammation in female C57BL/6 mice suffering from lipopolysaccharide (LPS)-induced ALI. We also demonstrated that OMZ-SPT can inhibit expression of the inflammatory cytokines tumor necrosis factor-α, interleukin-1β and interleukin-6 by ELISA in mice suffering from LPS-induced ALI and a mouse macrophage line (RAW264.7 cells). In addition, we showed that OMZ-SPT inhibited LPS-induced activation of nuclear factor-kappa B (NF-κB) signaling and total expression of NF-κB by western blotting. These data suggest that OMZ-SPT could be a novel therapeutic choice for ALI.

A new heteropolysaccharide was isolated from the fruiting bodies of Tricholoma matsutake which had a molecular weight of 12078 Da. The results of structural features analysis showed that T. matsutake polysaccharide, here named TMP-B, was mainly composed of α - D - glucose and α - D - galactose which ratios were 7:2 and had a backbone of 1, 4 - linked α - D - glucose which branches were mainly composed of two 6 - linked α - D - galactose residue, and the α - D - galactose was 1, 6 - linked. Antitumor activity results showed that heteropolysaccharide TMP-B could inhibit the growth of S180 tumor in vivo and promote the apoptosis of L929 cells in vitro. Immunoregulatory activity results showed that TMP-B could promote the proliferation of macrophages by affecting G0/G1 phase, S phases and G2/M phases and promote cytokines release and gene expression. The result of this study introduced Maerkang T. matsutake as a possible valuable source which helped to exhibit unique antitumor and immunoregulatory properties.

The antitumor and immunoregulatory activities of polysaccharide LC-1 from Lactarius camphoratum (Bull.) Fr.were studied by using the human cervical cancer Hela cells and the mouse macrophage RAW264.7 cells.The two kinds of cells were cultured in vitro, and the effects of different concentrations of LC-1 on the proliferation and cell cycle of Hela cells and the proliferation and phagocytosis of RAW264.7 cells were investigated.Moreover, the secretion of NO, IL-6 and TNF-alpha of RAW264.7 cells were tested using CCK-8 method, flow cytometry and ELISA technique, respectively.The results showed that polysaccharide LC-1 from L.camphoratum (Bull.) Fr.has obvious inhibitory effect on Hela cells in vitro, and could influence the apoptosis cell cycle of Hela cells.When the concentration of LC-1 was 10 μg/mL, the content of Sub peaks was 19.4%.It was further found that polysaccharide LC-1 could regulate the immune activity of macrophage.When the concentration of LC-1 was 10 μg/mL, the phagocytic activity and proliferation rate of macrophage were 67.48% and 87.09%, respectively.LC-1 could also promote the transformation of macrophages from G0/G1 phase to G2 phase and S phase and stimulate macrophages to produce IL-6 and TNF-α, showing a dose-dependent.However, no obvious effect on the production of NO was observed.In conclusion, in vitro, polysaccharide LC-1 from L.camphoratum (Bull.) Fr.could inhibit the growth of cervical carcinoma Hela cells and promote the proliferation and phagocytosis of macrophage, as well as stimulate macrophages to produce immune factors.

Background: The mechanism of the immunoregulatory activities of polysaccharide is still not clear. Materials and Methods: Here, we performed the B-cell, T-cell, and macrophage cell proliferation, the cell cycle analysis of macrophage cells, sequenced the transcriptomes of control group macrophages, and Boletus speciosus Frost polysaccharide (BSF-1) group macrophages using Illumina sequencing technology to identify differentially expressed genes (DEGs) to determine the molecular mechanisms of immunomodulatory activity of BSF-1 in macrophages. Results: These results suggested that BSF-1 could promote the proliferation of B-cell, T-cell, and macrophages, promote the proliferation of macrophage cells by abolishing cell cycle arrests in the G0/G1 phases, and promote cell cycle progression in S-phase and G2/M phase, which might induce cell division. A total of 12,498,414 and 11,840,624 bp paired-end reads were obtained for the control group and BSF-1 group, respectively, and they corresponded to a total size of 12.5 G bp and 11.8 G bp, respectively, after the low-quality reads and adapter sequences were removed. Approximately 81.83% of the total number of genes (8,257) were expressed reads per kilobase per million mapped reads (RPKM ≥1) and more than 1366 genes were highly expressed (RPKM >60) in the BSF-1 group. A gene ontology-enrichment analysis generated 13,042 assignments to cellular components, 13,094 assignments to biological processes, and 13,135 assignments to molecular functions. A Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that the mitogen-activated protein kinase (MAPK) signaling pathways are significantly enriched for DEGs between the two cell groups. Conclusion: An analysis of transcriptome resources enabled us to examine gene expression profiles, verify differential gene expression, and select candidate signaling pathways as the mechanisms of the immunomodulatory activity of BSF-1. Based on the experimental data, we believe that the significant antitumor activities of BSF-1 in vivo mainly involve the MAPK signaling pathways. Abbreviations used: BSF-1: Boletus speciosus Frost polysaccharide.

A new heteropolysaccharide was isolated from the fruiting bodies of Lactarius deliciosus Gray which had a molecular weight of 16 kDa and was mainly composed of the galactose and glucose. Structural elucidation results indicated that Lactarius deliciosus Gray polysaccharide (LDG-B) had a backbone of (1,6)-linked d-galactose and (1, 2, 6)-linked d-galactose which branches were mainly composed of 4-linked d-glucose and 6-linked d-galactose residue. Cell cycle test results showed that LDG-B could promote the proliferation of B cells and macrophage cells by affecting G0/G1 phase, S phases and G2/M phases. The analysis of transcriptomes sequence of macrophages showed a total of 1839 genes were identified as DEGs, and approximately 708 genes were up-regulated, whereas 1131 genes were down-regulated in LDG-B group. KEGG pathway enrichment analysis showed that the MAPK, JAK-STAT and NF-κB signaling pathways are significantly enriched for DEGs in LDG-B group. Analysis of transcriptome resources enabled us to examine gene expression profiles, verify differential gene expression, and select candidate signaling pathways as the mechanisms of the immunomodulatory activity of LDG-B.

The complementary DNA (cDNA) of the giant panda (Ailuropoda melanoleuca) ferritin light polypeptide (FTL) gene was successfully cloned using reverse transcription-polymerase chain reaction technology. We constructed a recombinant expression vector containing FTL cDNA and overexpressed it in Escherichia coli using pET28a plasmids. The expressed protein was then purified by nickel chelate affinity chromatography. The cloned cDNA fragment was 580 bp long and contained an open reading frame of 525 bp. The deduced protein sequence was composed of 175 amino acids and had an estimated molecular weight of 19.90 kDa, with an isoelectric point of 5.53. Topology prediction revealed one N-glycosylation site, two casein kinase II phosphorylation sites, one N-myristoylation site, two protein kinase C phosphorylation sites, and one cell attachment sequence. Alignment indicated that the nucleotide and deduced amino acid sequences are highly conserved across several mammals, including Homo sapiens, Cavia porcellus, Equus caballus, and Felis catus, among others. The FTL gene was readily expressed in E. coli, which gave rise to the accumulation of a polypeptide of the expected size (25.50 kDa, including an N-terminal polyhistidine tag).

A heteropolysaccharide was isolated from the fruiting bodies of Amanita caesarea using a diethylaminoethyl-cellulose column, Sephacryl S-300 gel column and Sephadex G-200 column. The Amanita caesarea polysaccharide was predominantly composed of -D-glucose and -D-lyxose at a ratio of 2:1, and it had a molecular weight of 19,329 Da. The structural features of the Amanita caesarea polysaccharide were investigated by a combination of total hydrolysis, methylation analysis, gas chromatography-mass spectrometry, and infrared spectra and nuclear magnetic resonance spectroscopy. The results showed that Amanita caesarea polysaccharide (termed AC-1) had a backbone of 1,4-linked -D-glucose and 1,3,6-linked -D-glucose, with branches of one 1-linked -D-lyxose residue. The antioxidant activity of AC-1 was evaluated by two biochemical methods, 2,2-azino-bis diammonium (ABTS(+)) radical scavenging activity and 1,1-diphenyl-2-picrylhydrazyl (DPPH-) radical scavenging activity. The uncontrolled production of free radicals is involved in various diseases, including cancer, atherosclerosis and degenerative aging processes. The results indicated that the Amanita caesarea polysaccharide exhibits strong antioxidant activity, thus, it may be a useful natural product antioxidant.

In the present study, we performed a proliferation assay, phagocytosis assay and cell cycle analysis of macrophages and sequenced the transcriptomes of control group macrophages and TMP-A group macrophages using Illumina sequencing technology to identify differentially expressed genes (DEGs) and determine the molecular mechanisms associated with differences in the immunomodulatory activity of TMP-A in macrophages. The results showed that TMP-A exhibits strong proliferation activity and phagocytosis activity in RAW264.7 cells in vitro and could also promote the proliferation of macrophage cells by abolishing cell-cycle arrest in the G0/G1 phase and promoting the cell cycle in the G2/M phase, which may induce cell division. A total of 12,616,096 and 11,798,839 bp paired-end reads were obtained for the control group and TMP-A group, respectively, and they corresponded to a total size of 12.5 G bp and 11.7 G bp, respectively, after the low-quality reads and adapter sequences were removed. Approximately 79.8% of the total number of genes (10,191) were expressed (RPKM ≥1), and more than 1,372 genes were highly expressed (RPKM >60) in the TMP-A group. A total of 1,043 unigenes were identified as DEGs, and approximately 486 genes were upregulated, whereas 557 genes were down-regulated, which might have contributed to the proliferation activity and phagocytosis activity of TMP-A in the RAW264.7 cells in vitro. A Gene Ontology (GO) enrichment analysis generated 13,042 assignments to cellular components, 13,094 assignments to biological processes, and 13,135 assignments to molecular functions. A KEGG pathway enrichment analysis showed that the MAPK and NF-κB signaling pathways are significantly enriched for DEGs between the two cell groups. Based on the experimental data, we believe that the significant antitumor activities of TMP-A in vivo involve the MAPK and NF-κB signaling pathways because the two signaling pathways intersect.

The sequences of the internal transcribed spacer (ITS) regions of ectomycorrhizal fungi collected from Sichuan Province were analyzed using a PCR primer pair specific to T. matsutake. The amplified fragments were sequenced and compared with each other to build a phylogenetic tree. The mRNA deep sequencing approach was adopted to identify differentially expressed T. matsutake genes among the transcriptomes developed from a Xiaojin sample. A phylogenetic analysis of the aligned sequences was performed using maximum-likelihood (ML) and neighbor-joining (NJ) analyses. The results clearly showed that the KD (KM657344) and BT (KM657342) strains were more closely related to each other than to other strains. Moreover, T. matsutake from Sichuan differed from those specimens derived from Heilongjiang, Yunnam, and Guizhou provinces of China, Finland, and Japan. Furthermore, there was extremely high homology among these T. matsutake samples, despite some genetic variation. In addition, the genome of T. matsutake was sequenced using Illumina sequencing technology (RNA-seq). In all, a total of 24,549,990 reads were obtained that yielded 18,266,492 high-quality clean reads. The quality reads were excluded later. The BLAST analysis of the sequence reads against the NR database indicated that T. matsutake shared a high number of contigs with Laccaria bicolor. The results also indicated that catalytic activity, metabolic processes, metabolic pathways, and biosynthesis of secondary metabolites were the main functions identified by gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Phylogenetic analysis showed that T. matsutake growing in Sichuan differed from samples growing in other regions. The differences in secondary metabolites between the Sichuan and Xiaojin samples may be due to differences in metabolic pathways. Thus the study provides a foundation for understanding T. matsutake biogeography and origins, and identifies DEGs in the Xiaojin sample to help elucidate the molecular mechanisms in secondary metabolite synthesis.

Background: Oligosaccharides are composed of a variable number of monosaccharide units and very important in the biologically diverse of biological systems. Materials and Methods: Crude water-soluble oligosaccharide was extracted from the fruiting bodies with water and then successively purified by DEAE-cellulose 52 and Sephadex G-100 column chromatography, yielding one major oligosaccharides fractions: LES-A. Structural features of Lactarius deliciosus (L. ex Fr.) Gray oligosaccharide (LDGO-A) were investigated by a combination of monosaccharide component analysis by thin layer chromatography, infrared spectra, nuclear magnetic resonance spectroscopy, scanning electron microscopy, and high-performance gel permeation chromatography analysis. Result: The results indicated that LDGO-A was composed of D-glucose and D-xylose, and the average molecular sizes was approximately 945 Da. The anti-tumor activity of LDGO-A was evaluated in vivo. The inhibitory rate in mice treated with 40 mg/kg LDGO-A can reach 40.02%, being the highest in the three doses, which may be comparable to mannatide. Histology of immune organs shows that the tissues arranged more regular and firmer, but the tumor tissue arranged looser in LDGO-A group than those in the control group. Meanwhile, there is no obvious damage to other organs, such as heart. The anti-tumor activity of the LDGO-A was usually believed to be a consequence of the stimulation of the cell-mediated immune response because it can significantly promote the lymphocyte and macrophage cells in the dose range of 100-400 g/mL in vitro. LDGO-A also effected the expression of some housekeeping genes mRNA in S180 tumor. Conclusion: Accordingly, the LDGO-A might serve as an effective healthcare food and source of natural anti-tumor compounds.

A novel heteropolysaccharide from the fruiting bodies of Gomphus clavatus Gray was isolated through Sephadex G-200 and DEAE-cellulose columns. The Gomphus clavatus Gray polysaccharide (GCG-1) was mainly composed of β-D-glucosepyranose (β-D-Glu) and α-D-galactopyranose (α-D-Gal) in a ratio of 3:2 and had a molecular weight of ~50,000 Da. The structure of GCG-1 was investigated by a combination of total hydrolysis, gas chromatography-mass spectrometry, methylation analysis, nuclear magnetic resonance spectroscopy and infrared spectra. The results indicated that GCG-1 had a backbone of (1 → 4)-β-D-glucosepyranose residues with branches at O-6 and the branches consisted of two with (1 → 3)-α-D-galactopyranose residue. Antioxidation test in vitro showed that it possessed strong free radical scavenging activity, which may be comparable to vitamin C and butylated hydroxytoluene. GCG-1 also induced the apoptosis of HepG-2 cells and affected the mRNA expression of various housekeeping genes in the HepG-2 cells. The results indicated that Gomphus clavatus Gray may be an ideal sources for antioxidant and anticancer agents.