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.

Cancer is one of the leading causes of mortality worldwide. Many of the protein from bacterium possess antitumor activity. The present study was carried out to evaluate the anti-cancer activity of Ribosomal Protein L36 (RPL36) from the Escherichia coli BL21 on human laryngeal carcinoma Hep-2 cells and Human hepatoma Hep G-2 cells. Evaluation of the effect of RPL36 response was made by the study of tumor growth response including cell morphology. The human laryngeal carcinoma Hep-2 cells treated with 0.3125-10 mu g/ml of RPL36 for 24 hours displayed significant cell growth inhibition (p < 0.05, n = 6) in assayed using MTT compared to the control (untreated) cells. For comparison, human hepatoma Hep G-2 cells displayed no significant change (p > 0.05; n = 6) when compared to the control (untreated) cells. The RPL36 has a time and dose dependent on Hep-2 cells growth inhibition. The data indicate that the effect at low concentrations is better than high concentrations, and the concentration of 0.625 1/4g/ml has the best rate of growth inhibition. The Real Time PCR array analysis of gene expression profile indicated that the anti-cancer activities of RPL36 on Hep-2 cells may involve DAPK1, DAPK3 and FADD signaling pathway for cell Apoptosis. The results of this study support the efficacy of Escherichia coli BL21ribosomal protein L36 as an anticancer agent for human laryngeal carcinoma Hep-2 cells. However, further studies of the mechanism and the signal transduction pathways are required.

The fungal polysaccharides have been revealed to exhibit a variety of biological activities, including antitumor, immune-stimulation and antioxidation activities. In the present study, the immune and anticancer activities of a novel polysaccharide, BSF-A, isolated from Boletus speciosus Frost was investigated. The inhibitory rate of S180 tumors in mice treated with 40 mg/kg BSF-A reached 62.449%, which was the highest rate from the three doses administered; this may be comparable to mannatide. The antitumor activity of BSF-A is commonly considered to be a consequence of the stimulation of the cell-mediated immune response, as it may significantly promote the macrophage cells in the dose range of 100-400 µg/ml in vitro. The levels of the cytokines, IL-6, IL-1β and TNF-α, and nitric oxide, induced by BSF-A treatment at varying concentrations in the macrophage cells were similar to the levels in the cells treated with lipopolysaccharide. There was weak expression of the TNF-α, IL-6, IL-1β and inducible nitric oxide synthase mRNA in the untreated macrophages, but this increased significantly in a dose-dependent manner in the BSF-A-treated cells. BSF-A also had a time- and dose-dependent effect on the growth inhibition of the Hep-2 cells, with the concentration of 400 µg/ml having the highest inhibitory rate. A quantitative PCR array analysis of the gene expression profiles indicated that BSF-A had anticancer activities that affected cell apoptosis in the Hep-2 cells. The results obtained in the present study indicated that the purified polysaccharide of Boletus speciosus Frost is a potential source of natural anticancer substances.

In the present study, morphological and genetic characterization of the Tricholoma matsutake was identified from the fruiting bodies of Xiaojin Tricholoma matsutake in southwestern China. The morphological character of Xiaojin sample is coincident with the character of Tricholoma matsutake genus and a phylogenetic tree was also generated based on the sequences of the internal transcribed spacer (ITS) region of related fungi. The analysis of ITS sequence showed that fungus was clustered together with Tricholoma matsutake (AF367418.1) in the phylogenetic tree, which indicated that fungus was a kind of fungi belonging to Subgenus Tricholoma. Meanwhile, the improved cPDA medium was identified as the most suitable medium components for solid culture. Orthogonal design was used to test the relative importance of various factors on liquid fermentation. Among the variables, rotary speed found to be the most significant variables. From further optimization studies, the optimized values of the variables for production were as follows: pH 5.5, temperature 20 degrees C, speed 200 r/min, inoculation volume 20% and vaccination age 5 d. In the study, the highest production of 33.365 g/l mycelium dry weight was obtained with the production of 0.4958248 g/l total sugar. The polysaccharide isolated from the mycelium of Xiaojin Tricholoma matsutake showed relative strong antioxidant activity using DPPH-radical scavenging method. The antioxidant activity for DPPH-radical and the concentration of the T-1 was positively correlated.

In this study, a heteropolysaccharide was isolated from the fruiting bodies of Lactarius camphoratum (Bull.) Fr. through DEAE-cellulose column and Sephadex G-200 column. The L. camphoratum (Bull.) Fr. polysaccharide (LC-1) had a molecular weight of 9279 Da and was mainly composed of β-D-Glu and α-D-Gal which ratios were 2:1. Structural features of L. camphoratum (Bull.) Fr. polysaccharide (LC-1) were investigated by a combination of total hydrolysis, methylation analysis, gas chromatography-mass spectrometry (GC-MS), infrared (IR) spectra and nuclear magnetic resonance (NMR) spectroscopy. The results indicated that L. camphoratum (Bull.) Fr. polysaccharide (LC-1) had a backbone of (1→4)-β-D-glucopyranose residues which branches at O-6 based on the experimental results. The branches were mainly composed of one with →3)-α-D-galactopyranose residue. The antioxidant activity of LC-1 was evaluated with two biochemical methods, including 1,1-diphenyl-2-picrylhydrazyl (DPPH(-)) radical scavenging, scavenging activity of 2,2'-azino-bis(3-ethylbenzthiazoline-6-suphonic acid) diammonium (ABTS(+)) radical. The results indicated that LC-1 showed strong antioxidant.

从猴头菇(Hericium erinaceus)子实体中,经过除杂、粉碎、热水浸提、醇沉处理后,得到了猴头菇杂聚糖粗提物,并运用DEAE-cellulose column进行柱层析分离纯化,得到猴头菇寡聚糖(HEO-A).通过水解,单糖分析,扫描电镜技术(SEM),高效凝胶渗透色谱(HPGPC),红外光谱技术(IR)和核磁共振技术(NMR)对猴头菇寡聚糖进行了初步的结构分析.结果显示,猴头菇寡聚糖由木糖和葡萄糖两种单糖构成,均重分子量为1877D.在体外化学模拟条件下,研究了不同浓度的猴头菇寡聚糖对羟基自由基(·OH)、1,1-二苯基-2-苦苯肼自由基(DPPH·)及ABTS自由基的清除作用.对DPPH·自由基的清除能力,其IC50值为12.5mg/mL;对·OH自由基的清除能力,当HEO-A浓度达到5mg/mL时,其清除率可达到最大(37.51%);对ABTS自由基的能力,其IC50值为0.93mg/mL,且当HEO-A的浓度为1.5mg/mL时,清除率即可达95.44%.这些结果说明猴头菇寡聚糖(HEO-A)具有显著的抗氧化功能,可以作为一种理想的天然抗氧化剂资源.

Ribosomal protein S23 (RPS23) is a component of the 40S small ribosomal subunit encoded by the RPS23 gene, which is specific to eukaryotes. The cDNA and genomic sequence of RPS23 were cloned from Ailuropoda melanoleuca (A. melanoleuca) using reverse transcription‑polymerase chain reaction (RT-PCR) technology and touchdown PCR, respectively. The two sequences were analyzed preliminarily and the cDNA of the RPS23 gene was overexpressed in Escherichia coli (E. coli) BL21. The cDNA of RPS23 cloned from giant panda was 472 bp, and it contained an open reading frame (ORF) of 432 bp encoding 142 amino acids. The nucleotide sequence of the coding sequence showed a high degree of homology to some mammals as determined by BLAST analysis, similar to the amino acid sequence. The genomic sequence was 2,105 bp in length, with 4 exons and 3 introns. The primary structure analysis revealed that the molecular weight of the putative RPS23 protein was 15.80 kDa with a theoretical isoelectric point (pI) of 11.23. The molecular weight of the recombinant protein RPS23 was 21.5 kDa with a theoretical pI of 10.57. Topology prediction showed that there are seven different patterns of functional sites in the RPS23 protein of giant panda. RPS23 was successfully expressed in E. coli and its protein fused with the N‑terminal His‑tagged protein triggered the accumulation of an expected 21.5‑kDa polypeptide. The inhibitory rate of tumor growth in mice treated with 0.1 µg/ml RPS23 protein was 49.45%, the highest in the three doses used, which may be comparable to mannatide treatment. Histology of immune organs showed that the tissues were characterized by a regular and tight arrangement, while tumor tissues of the mice in the RPS23 group exhibited a loose arrangement compared to the control group. However, there was no obvious damage to other organs, such as the heart, lung and kidney. Investigations are currently being conducted to determine the bioactive principles of the recombinant protein RPS23 responsible for its anticancer activity.