Over-expression of lncRNA TMEM161B-AS1 promotes the malignant biological behavior of glioma cells and the resistance to temozolomide via up-regulating the expression of multiple ferroptosis-related genes by sponging hsa-miR-27a-3p.

A growing body of evidence suggests that long-chain non-coding RNA (lncRNA) plays an important role in the malignant biological behavior and drug resistance of glioblastoma (GBM) cells. In this study, we analyzed the role and potential mechanism of lncRNA TMEM161B-AS1 in the malignant biological behavior of GBM cells and temozolomide (TMZ) resistance. Studies have found that FANCD2 and CD44 are significantly related to the occurrence of GBM, TMZ resistance and the survival of GBM patients. Knockdown of TMEM161B-AS1 down-regulated the expression of FANCD2 and CD44 by sponging hsa-miR-27a-3p, inhibited the proliferation, migration, invasion and promoted apoptosis, ferroptosis of U87 cells and U251 cells. Down-regulation of lncRNA TMEM161B-AS1 and/or over-expression of hsa-miR-27a-3p down-regulated the expression of FANCD2 and CD44, and inhibited the tumor growth in nude mice. These results demonstrated that the lncRNA TMEM161B-AS1-hsa-miR-27a-3p-FANCD2/CD44 signal axis regulated the malignant biological behavior of GBM and TMZ resistance. These findings were expected to provide promising therapeutic targets for the treatment of glioma.

Polysaccharides from Premna microphylla turcz ameliorate inflammation via the enhancement of intestinal resistance in host.

ETHNOPHARMACOLOGICAL RELEVANCE: Premna microphylla turcz is traditionally used as a folk remedy. Its roots, stems and leaves can be invoked as medicines, which have the functions of detoxification, swelling and hemostasis. It belongs to the Premna in the Verbenaceae and is mainly distributed in the mountains of southeastern China. However, there are few reports of in-depth studies on the anti-inflammatory effects of polysaccharide, which was the main component in Premna microphylla turcz. MATERIALS AND METHODS: The flies were fed with standard corn flour-yeast medium to cause inflammation by sodium lauryl sulfate (SDS). The treatment group contained Premna microphylla turcz polysaccharide (pPMTLs) extract. The survival rate was obtained by feeding a vial containing five layers of filter paper, which was infiltrated with the 5% sucrose solution contaminated with SDS or SDS polysaccharide. The microvilli and nucleus of the midgut epithelial cells of different treatments were observed by transmission electron microscope, and the expression of inflammation-related genes was detected by real-time quantitative PCR (qRT-PCR). Finally, 16S rDNA analysis was conducted on the differences in the composition of the intestinal microbes of Drosophila. RESULTS: In the current study, we showed that pPMTLs significantly prolonged the life span of SDS-inflamed flies from 5 days to 6 days. And pPMTLs reduced the rupture of microvilli in the midgut and restored the nuclear structure. In addition, pPMTLs significantly improved expression level of immune-related genes in Inflammation Drosophila especially the defensin (4.32 ± 0.75 vs 9.97 ± 0.52 SDS-polysaccharide group: SDS group, p < 0.001). The analysis of intestinal microbiota showed that pPMTLs decreased the relative abundance of Raoultella while Wolbachia increased (p < 0.05). CONCLUSIONS: Collectively, our results revealed the potential application of pPMTLs in enhancing inflammation defense, which would be enormous significance for the inflammation-related disorders treatment.

The level of thymic stromal lymphopoietin and its gene polymorphism are associated with rheumatoid arthritis.

OBJECTIVE: To study the correlation between TSLP gene SNPs and RA in a Han Chinese population. METHODS: The genotypes of TSLP genes rs11466749, rs11466750 and rs10073816 among 197 RA patients and 197 controls were analysed by direct sequencing. ELISA was used to detect the plasma TSLP level. Logistic regression analysis was also conducted to identify risk factors for RA. RESULTS: The rs11466749 locus GG genotype (OR = 5.30, 95% CI: 1.76-15.95, P < 0.01), dominant model (OR = 1.68, 95% CI: 1.03-2.73, P = 0.04), recessive model (OR = 5.15, 95% CI: 1.72-15.43, P < 0.01), and G allele (OR = 2.02, 95% CI: 1.33-3.09, P < 0.01) were associated with an increased risk of RA. The rs1073816 locus AA genotype (OR = 4.58, 95% CI: 1.49-14.01, P < 0.01), dominant model (OR = 1.75, 95% CI: 1.09-2.79, P = 0.03), recessive model (OR = 4.27, 95% CI: 1.40-13.00, P = 0.03) and A allele (OR = 1.94, 95% CI: 1.29-2.91, P < 0.01) were associated with an increased risk of RA. The rs1073816 locus GA genotype (OR = 0.29, 95% CI: 0.18-0.45, P < 0.01), dominant model (OR = 0.32, 95% CI: 0.21-0.49, P < 0.01) and A allele (OR = 0.45, 95% CI: 0.32-0.63, P < 0.01) were related to a decreased risk of RA susceptibility. The rs1466749 locus GG genotype, rs11466750 AA genotype, and rs10073816 GG genotype were independent risk factors for RA (P < 0.05). The AUC of plasma TSLP level in the diagnosis of RA was 0.8661 (95% CI: 0.8301-0.9002, P < 0.001). There were statistically significant differences in plasma TSLP levels among subjects with different genotypes at rs11466749, rs11466750, and rs10073816 in the TSLP gene (P < 0.05). CONCLUSION: Plasma TSLP levels are a potential molecular marker of RA. SNPs at rs11466749, rs11466750 and rs10073816 of the TSLP gene are related to the susceptibility of the Han Chinese population to RA.

Preparation and characterisation of a novel hydrogel based on Auricularia polytricha ß-glucan and its bio-release property for vitamin B12 delivery.

BACKGROUND: This study investigates a novel hydrogel synthesis method and its bio-release property. This hydrogel, with a three-dimensional network structure based on Auricularia polytricha ß-glucan, was characterised by means of Fourier transform infrared spectroscopy, 1 H NMR and scanning electron microscopy. Vitamin B12 (VB12 , cobalamin) as a hydrophilic functional food component was entrapped into these hydrogels. The in vitro release profile of VB12 was established in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). RESULTS: The results showed that the hydrogel had medium pore size from 30 to 300 µm, and the swelling ratio increased with the degree of substitution. The hydrogel demonstrated good stability in SGF and bio-release capability in SIF for VB12 . The accumulated release rate is about 80% in SIF and below 20% in SGF, which indicated the significant different release property in stomach and intestine. CONCLUSION: The Auricularia polytricha ß-glucan-based hydrogel has a good swelling ratio, pepsin stability and pancrelipase-catalysed biodegradation property. The bio-release rate is significantly different in SIF and SGF, which indicated that this hydrogel could be a good intestinal target carrier of VB12 . © 2017 Society of Chemical Industry.

Structural characterization and immunomodulatory activity of a novel polysaccharide from Pteridium aquilinum.

PAP1-A, a novel heteropolysaccharide with an average molecular mass of 1.35×105Da, was isolated from Pteridium aquilinum using a combination of chromatography by DEAE Sepharose Fast Flow and Sepharose 4B. The monosaccharide component of PAP1-A was L-rhamnose, L-arabinose, L-fucose, D-xylose, D-mannose, D-glucose and D-galactose in the molar ration of 1.82: 1.53: 1.42: 1.31: 5.24: 1: 12.35. The predicted structure of PAP1-A was established to be a complex polysaccharide with a main chain consisted of α-(1→4)-linked galactose partially substituted at O-6 position, with the (1→2)-linked xylose, (1→3)-linked arabinose, (1→3)-linked rhamnose, (1→3,6)-linked mannose, and (1→6)-linked mannose, as branches. Fucose, glucose, mannose, and rhamnose were located at the termini of the branches. The immunomodulatory activity assay showed that PAP1-A could significantly promote the RAW264.7 cells proliferation and induce the production of NO from RAW264.7 cells. Therefore, PAP1-A shows as a potent immunomodulator with potential applications in the medical and food industries.

Preparative separation of conjugated linoleic acids (CLAs) from fermented Camellia oleifera Abel cake by ß-cyclodextrin (ß-CD) encapsulation using pH-zone-refining countercurrent chromatography.

This paper concentrates on the separation of three conjugated linoleic acid (CLA) isomers (trans-9,trans-11 CLA, trans-10,cis-12 CLA and cis-9,trans-11 CLA) by ß-cyclodextrin (ß-CD) encapsulation using countercurrent chromatography from Camellia oleifera Abel cake fermented by lactic acid bacteria Lactobacillus sp. LL-ZSDS001. The elution sequence of the CLA isomers, the mixing zones and mechanism of separation are discussed. The separation of 305.9mg of the crude sample yielded three isomeric compounds: 91.3mg of trans-9,trans-11 CLA, 84.1mg of trans-10,cis-12 CLA and 79.7mg of cis-9,trans-11 CLA at high purities of 98%, 94% and 96%, respectively.

Structure analysis and antimutagenic activity of a novel salt-soluble polysaccharide from Auricularia polytricha.

BACKGROUND: Auricularia polytricha is known to be a highly nutritious foodstuff. We report here the purification, structure characterization and antimutagenic activity in vivo of a 0.9% NaCl solution-soluble polysaccharide (SSP) from the mycelia of A. polytricha. RESULTS: Analysis by high-performance liquid chromatography coupled to a TSK-G5000PWXL column and gel filtration chromatography on Sephacryl S-400 HR indicated that SSP is homogeneous with an average molecular weight of about 9.30 × 10(5) Da. The structure of SSP was revealed by chemical methods, Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. Results indicated that SSP is a glucan consisting of a1,3-ß-glucan, 1,6-α-glucan, 1,4-α-glucan and 1,3-α-glucan backbone with a single 1,6-α-d-glucopyranosyl side-branching unit on every nine residues, on average, along the main chain. Atomic force microscopy indicates the presence of macromolecular species in morphology and shows a clear association of prolate particle. Meanwhile, SSP was found to significantly preventing micronuclei in polychromatic erythrocytes and reticulocytes of mice (P < 0.01). CONCLUSION: These results indicate that polysaccharide SSP from A. polytricha exhibits antimutagenic activity against the in vivo DNA-damaging effect of the indirectly acting alkylating agent cyclophosphamide.

Structure characterisation of a α ß-glucan polysaccharide from Auricularia polytricha.

A water-soluble α ß-glucan with a molecular weight of 1.62 × 10(5) Da, AAPS -1, was obtained from the fruiting bodies of the edible mushroom Auricularia polytricha by a combined separation of high-speed countercurrent chromatography (HSCCC)-Sephacryl S-300 HR column chromatography. The structure of AAPS-1 was elucidated that the polysaccharide possesses a backbone composed of (1 → 6)-linked-α-D-glucopyranosyl and (1 → 6)-linked-ß-D-glucopyranosyl residues, partially substituted at O-3 of ß-D-glucopyranosyl residue by side chain (1 → 4)-α-D-Glcp and terminated with non-reducing end α-D-Glcp-(1 → on the basis of the analyses of partial hydrolysis, periodate oxidation, acetylation, methylation and NMR spectroscopy ((1)H, (13)C).

Isolation of a polysaccharide with anticancer activity from Auricularia polytricha using high-speed countercurrent chromatography with an aqueous two-phase system.

Polysaccharides from a crude extract of Auricularia polytricha were separated by high-speed countercurrent chromatography (HSCCC). The separation was performed with an aqueous two-phase system of PEG1000-K2HPO4-KH2PO4-H2O (0.5:1.25:1.25:7.0, w/w). The crude sample (2.0 g) was successfully separated into three polysaccharide components of AAPS-1 (192 mg), AAPS-2 (137 mg), and AAPS-3 (98 mg) with molecular weights of 162, 259, and 483 kDa, respectively. These compounds were tested for growth inhibition of transplanted S180 sarcoma in mice. AAPS-2 had an inhibition rate of 40.4%. The structure of AAPS-2 was elucidated from partial hydrolysis, periodate oxidation, acetylation, methylation analysis, and NMR spectroscopy (1H, 13C). These results showed AAPS-2 is a polysaccharide with a backbone of (1-->3)-linked-beta-d-glucopyranosyl and (1-->3, 6)-linked-beta-D-glucopyranosyl residues in a 2:1 ratio, and has one terminal (1-->)-beta-D-glucopyranosyl at the O-6 position of (1-->3, 6)-linked-beta-D-glucopyranosyl of the main chain.

Completed preparative separation of alkaloids from Cephaltaxus fortunine by step-pH-gradient high-speed counter-current chromatography.

Cephalotaxine-type alkaloids are the anti-cancer components in twigs, leaves, roots and seeds of Cephalotaxus fortunine. It is very important to use the limited resource by finding an efficient purification technology of the alkaloids. Separation of cephalotaxine-type alkaloids in Cephalotaxus fortunine by step-pH-gradient high-speed counter-current chromatography (step-pH-gradient HSCCC) was studied in this paper. The step-pH-gradient HSCCC was performed on a HSCCC instrument equipped with a 400-mL column, using the upper phase of ethyl acetate-n-hexane-water, with added 0.01% trifluoroacetic acid (TFA) as stationary phase, and the lower phase of ethyl acetate-n-hexane-water, with added 2% NH(4)OH, 0.2% NH(4)OH and 0.05% TFA as mobile phase. For each separation, 800mg of extract of cephalotaxine-type alkaloids was separated to yield 9.3mg of drupacine, 15.9mg of wilsonine, 130.4mg of cephalotaxine, 64.8mg of epi-wilsonine, 12.8mg of fortunine and 35.6mg of acetylcephalotaxine with purities 81.2%, 85.7%, 95.3%, 97.5%, 89.1% and 96.2%, respectively. The recovery of each alkaloid was more than 90%. The structures of the six alkaloids were identified by electrospray ionization mass spectrum (ESI-MS) and (1)H and (13)C NMR.