Rui Li

Soochow University (PRC), Wu-hsien, Jiangsu Sheng, China

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Publications (21)43.89 Total impact

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    ABSTRACT: A number of single-nucleotide polymorphisms within the 3'-UTR of genes have been shown to relate to the occurrence of cancers. In this study, by using polymerase chain reaction-restriction fragment length analysis method, we determined an SNP rs1599795 in the 3'-UTR of CD80 gene in 183 gastric cancer patients and 348 healthy controls. Statistical analysis results showed that SNP rs1599795 genotypes were significantly correlated with the risk of gastric cancer. Compared with the AA homozygotes, the TA heterozygotes were significantly more prevalent in the patients (OR 1.44, 95 % CI 0.98-2.11) with a larger tumor size (P = 0.001), deeper infiltration (P = 1.5 × 10(-5)), higher possibility of lymph node metastasis (P = 0.003), and more in the late stage (TNM stage III and IV; P = 0.003); the TT homozygotes had larger tumor size (P = 0.001) and lower degree of differentiation (P = 2.2 × 10(-4)). Dual-luciferase reporter assays showed that miR-132-3p, miR-212-3p, and miR-361-5p inhibited the expression of CD80 through binding with the CD80 3'-UTR, and this inhibitory role of miR-132-3p, miR-212-3p, and miR-361-5p was impacted by rs1599795. Our findings have shown that the SNP rs1599795 in CD80 3'-UTR, through disrupting the regulatory role of miR-132-3p, miR-212-3p, and miR-361-5p in CD80 expression, contributed to the occurrence of gastric cancer.
    Medical oncology (Northwood, London, England). 08/2014; 31(8):60.
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    ABSTRACT: Co-inhibitor B7-H1 expresses in various cancers and contributes to cancer immune evasion by inhibiting T cell activation and proliferation, yet the regulatory mechanisms for B7-H1 over-expression in cancers remain largely unknown. Here, the expression of B7-H1 and PTEN proteins were firstly detected by using immunohistochemistry method. B7-H1 immunoreactivities were found in 54.5% (55/101) of the colorectal cancer tissues with no expression in the normal tissues, and the PTEN protein immunoreactivities were observed in 51.5% (52/101) of the colorectal cancer tissues and 72.3% (73/101) of the normal tissues. Statistical analysis results indicated that the B7-H1 expression was negatively correlated to the PTEN expression in colorectal cancer (p=0.001). Then the expressions of microRNAs (miRNAs) in six pairs of colorectal cancer and normal tissues were determined by miRNA array, and 30 up-regulated miRNAs were found in the colorectal cancer tissues. Finally, the impact of these up-regulated miRNAs on PTEN expression was tested by using dual-luciferase reporter assay system, from which the results indicated that miR-20b, -21, and -130b were involved in suppression of PTEN expression. These findings suggest that miR-20b, -21, and -130b, up-regulated in colorectal cancer, through inhibiting the expression of PTEN, result in B7-H1 over-expression in colorectal cancer.
    Human immunology 01/2014; · 2.55 Impact Factor
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    ABSTRACT: A number of single nucleotide polymorphisms (SNPs) within the 3'-UTR of genes have been proved to be contributed to the risk of cancers. Here, we determined an SNP rs4819388 in the 3'-UTR of B7-H2 gene in 183 gastric cancer patients and 348 healthy controls by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Statistical analysis results showed that the rs4819388 genotypes were significantly related to the occurrence of gastric cancer. Compared with the GG homozygotes, the GA heterozygotes were significantly more prevalent in the patients (OR=1.60, 95%CI=1.08-2.37). In addition, the A allelic frequencies were significantly higher than that of the G allele in the patients with lymphatic metastasis (p=0.034) and/or advanced TNM stage (p=0.032). Dual-luciferase reporter assays showed that miR-24 inhibited the expression of B7-H2 through binding with the B7-H2 3'-UTR, and this inhibitory role of miR-24 was impacted by rs4819388. Our findings suggest that the SNP rs4819388 in B7-H2 3'-UTR, through disrupting the regulatory role of miR-24 on B7-H2 expression, contributes to the occurrence of gastric cancer.
    Molecular Immunology 05/2013; 56(1-2):98-103. · 2.65 Impact Factor
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    ABSTRACT: This study is designed to screen the CD40 related signal transduction pathway in AGS cells and construction of gene silencing vector. Analysis results showed 414 differential genes expression, including upregulation of 209 genes and downregulation of 205 genes. Basing on the ratio of signal in experimental group to signal in control group, 45 genes (38 genes upregulation and seven genes downregulation) with significant (P < 0.01) change in expression levels were screened according to the screening standard (signal log ratio ≥1 or ≤-1). These genes involved into metabolism, cell cycle and apoptosis, signal transduction and stress response. Furthermore, PI3K mRNA expression level in PI3K siRNA transfected AGS cells was 0.2335 ± 0.0116 72 h after transfection. This value was significantly (P < 0.05) lower than that in blank and negative control groups. PI3K protein expression in PI3K siRNA transfected AGS cells was significantly (P < 0.05) lower than that in blank and PI3K siRNA/N transfected groups. Therefore, PI3K siRNA gene silencing vector can significantly inhibit PI3K mRNA and protein expression in AGS cells.
    Molecular Biology Reports 05/2013; · 2.51 Impact Factor
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    ABSTRACT: Single nucleotide polymorphisms (SNPs) in putative microRNA (miRNA) target sites (miRSNPs) could affect the binding of miRNA with the target and contribute to the susceptibility of human cancers. However, the role of miRSNPs in gastric cancer susceptibility remains largely unknown. Since the over-expression of B7-H1 protein has been reported to be closely related to disease progression of gastric cancer, we investigated the possible role of miRSNPs at the 3'-untranslated region (3'-UTR) of B7-H1 in the risk of developing gastric cancer. In this association study on 205 gastric adenocarcinoma patients and 393 non-cancer controls, we found that the genotype distribution of a common C>G polymorphism (rs4143815) was significantly different between the cases and controls (P = 1.32 × 10(-8)). Compared with CC homozygotes, GG homozygotes and G allele carriers showed 3.73-fold (P = 2.98 × 10(-8)) and 1.85-fold (P = 0.002) increased risk of gastric adenocarcinoma, respectively. Stratified analyses indicated that variant genotypes had a strong association with the clinic-pathological features of gastric cancer including differentiation grade, depth of tumor infiltration, and tumor node metastasis (TNM) stage (P < 0.001). Luciferase reporter assay indicated that this SNP might be responsible for aberrant B7-H1 protein expression in gastric cancer by disrupting the interaction between miR-570 and B7-H1 mRNA. These results are consistent with our hypothesis and indicate that genetic polymorphisms influencing B7-H1 expression modify cancer susceptibility.
    Human Genetics 02/2013; · 4.63 Impact Factor
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    ABSTRACT: Background: To better understand gastric cancer occurrence and prognosis, we explored the expression of molecules in the CD40 pathway and their correlation with gastric cancer prognosis. Patients and Methods: We measured the expression of CD40, VEGF, AKT, PI3K, and S100 in gastric cancer tissues and adjacent normal tissues from 128 patients by immunohistochemistry. Results: The expression of CD40, VEGF, AKT, and PI3K were significantly higher in tumor tissue than in normal tissue, while S100 expression in dendritic cells (DC) was lower. Expression of CD40, VEGF, AKT, and PI3K significantly increased with T stage, while S100 expression decreased with T stage. Lymph node metastasis was associated with low or negative S100 expression. PI3K expression increased with clinical stage, while negative S100 expression was associated with higher clinical stages. Multivariate analysis did not indicate significant associations between any of these markers and recurrence or mortality. Conclusion: The correlation between T stage of gastric cancer and the higher expression of CD40, VEGF, AKT, and PI3K, along with lower S100 expression in DC, may provide insights into future targets for more effective immunotherapy for cancer.
    Onkologie 01/2013; 36(1-2):26-31. · 1.00 Impact Factor
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    ABSTRACT: In this study, we investigate effect of PI3K gene silencing on growth, migration and related proteins expression of CD40 signal-mediated gastric cancer cells. We observed that combination of sCD40L with PI3K siRNA could significantly inhibit AGS cells growth, block cells in G1 phase, and promote tumour cells apoptosis after 24 h treatment. Transwell test showed that numbers of cells per visual field in group PI3K siRNA or group sCD40L (after 24 h PI3K siRNA or sCD40L alone treatment) were fewer than that (32.54 ± 4.22) in control group. Numbers of cells per visual field in (after 24 h combination treatment of PI3K siRNA with sCD40L) were significantly fewer than that in group PI3K siRNA or group sCD40L. Compared with group sCD40L, expression level of Fas protein in group sCD40L + PI3K siRNA was significantly increased. The findings suggest that PI3K siRNA may strengthen CD40-induced specific antitumour effect via blocking PI3K/Akt signal pathway, resisting tumour immunoediting regulated by CD40 signal. Combination of sCD40L and PI3K siRNA is an important mechanism of gastric cancer therapy.
    Molecular Biology Reports 11/2012; · 2.51 Impact Factor
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    ABSTRACT: Allele-specific amplification on the basis of polymerase chain reaction (PCR) has been widely used for single-nucleotide polymorphism (SNP) genotyping. However, the extraction of PCR-compatible genomic DNA from whole blood is usually required. This process is complicated and tedious, and is prone to cause cross-contamination between samples. To facilitate direct PCR amplification from whole blood without the extraction of genomic DNA, we optimized the pH value of PCR solution and the concentrations of magnesium ions and facilitator glycerol. Then, we developed multiplex allele-specific amplifications from whole blood and applied them to a case-control study. In this study, we successfully established triplex, five-plex, and eight-plex allele-specific amplifications from whole blood for determining the distribution of genotypes and alleles of 14 polymorphisms in 97 gastric cancer patients and 141 healthy controls. Statistical analysis results showed significant association of SNPs rs9344, rs1799931, and rs1800629 with the risk of gastric cancer. This method is accurate, time-saving, cost-effective, and easy-to-do, especially suitable for clinical prediction of disease susceptibility.
    Genetic Testing and Molecular Biomarkers 10/2012; · 1.44 Impact Factor
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    ABSTRACT: The objective of the paper is to investigate the combined effect of sCD40L and phosphatidylinositol 3-kinase (PI3K) siRNA on transplanted tumours growth and microenvironment in nude mice with gastric cancer. 48 h after modeling, the animals were randomly divided into saline + AGS group (A), sCD40L + AGS group (B), saline + PI3K siRNA group (C) and sCD40L + PI3K siRNA group (D), six mice in each group. The mouse in the groups was inoculated with exponential phase AGS cell or PI3K gene silencing cells (100 μl, 5 × 10(6)). After tumour size reaches 0.2-0.3 cm, Tumours in animals of groups were injected with sCD40L (100 μl, 10 mg/kg) or equal volume of saline, thrice each day, respectively. Microvessel density (MVD), apoptosis index, and expression levels of PI3K, Survivin and vascular endothelial growth factor (VEGF) proteins in transplanted tumor cells in gastric cancer nude mice were analyzed by utilizing Immunohistochemistry, western blot, terminal deoxynucleotidyl transferase dUTP nick end labeling assays. Results showed that combination of sCD40L with PI3K siRNA could significantly decrease tumour size, MVD, expression levels of PI3K, Survivin and VEGF proteins, and increase apoptosis index. It can be concluded that combination of sCD40L with PI3K siRNA provides a promising future for gastric cancer therapy.
    Molecular Biology Reports 06/2012; 39(9):8755-61. · 2.51 Impact Factor
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    ABSTRACT: CD40 signaling plays a critical role in the survival rate of gastric cancer patients. Tumour samples were collected from 73 patients with who were diagnosed as gastric cancer in general surgery department in the 1st affiliated hospital of Suzhou University between September 2002 and July 2003. All patients had not received radiotherapy and chemotherapy before operation. These patients include 46 male and 27 female. Here we show that CD40 is constitutively expressed in the human gastric carcinoma tissues, and CD40 protein and mRNA positive expression in gastric cancer tissues closely correlated with lymph node metastasis and tumour TNM stage. CD40 positive expression in gastric cancer patients with lymph node metastasis was markedly higher than that in gastric cancer patients without lymph node metastasis. CD40 positive expression in stage III-IV gastric cancer patients was markedly higher than that in stage I-II gastric cancer patients. Moreover, CD40 expression closely correlated with prognosis of gastric cancer patients. Therefore, CD40 was taken as grouping variable, and lymph node metastasis and clinical staging were taken as stratification variables, respectively, further analysis showed that prognosis in gastric cancer patients with lymph node metastasis and CD40 positive expression was markedly worse than that in gastric cancer patients without lymph node metastasis and CD40 negative expression (P = 0.0076). These results suggest that CD40 signaling plays a critical role in the survival of gastric cancer patients.
    Molecular Biology Reports 06/2012; 39(9):8741-7. · 2.51 Impact Factor
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    ABSTRACT: The aim of this study was to investigate the molecular mechanisms underlying the antitumour effects of CD40L through analysing the change of genes expression profile in AGS using Affymetrix Gene Chip. Human gastric carcinoma AGS cells were first incubated with 2 μg/ml sCD40L or equal volume of medium (control) in F12 medium. RNA was isolated from AGS and were reverse transcribed, labeled with digoxigenin-11-dUTP, and then hybridized with Clontech Atlas mouse cDNA expression arrays for comparison. Performing clustering analysis, we found that 7 detected genes were down-regulated and 38 were upregulated as the sCD40L acted on AGS. To further verify the results of gene chip screening, Gene Database was searched, finding that the most significantly up-regulated genes were Gadd45a, c-Jun and Bcl-2, and the most significantly down-regulated genes were Cyclin D1, CDC6, TNFR10B, c-IAP2 and ORC5L. Based upon these findings, the signalling pathways that possibly mediate CD40-induced apoptosis are proposed.
    Molecular Biology Reports 02/2012; 39(6):6615-23. · 2.51 Impact Factor
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    ABSTRACT: Inhibitory costimulatory molecule CD274 expresses in various cancers and contributes to cancer immune evasion by inhibiting T cell activation and proliferation, yet the regulatory mechanisms for CD274 overexpression in cancers are poorly understood. In this study, we discovered a novel mechanism of CD274 expression regulated by miR-570. A guanine-to-cytosine mutation at the 3'-UTR of CD274 mRNA led to CD274 overexpression by disrupting the miR-570 binding. The mutations were widely observed in cancers by sequencing of 276 gastrointestinal cancers (esophageal, gastric, colorectal, hepatocellular, and pancreatic cancers). This mutation was significantly associated with CD274 overexpression in gastric cancer (P = 1.44×10(-10)) and with the pathological features including differentiation grade, depth of tumor invasion, lymph node metastasis, and tumor-node-metastases (TNM) stage. These findings suggest a novel regulatory mechanism for CD274 overexpression in gastric cancer mediated by miR-570 and a somatic mutation in CD274 3'-UTR, and provide a new insight to gastric carcinogenesis.
    Human Mutation 12/2011; 33(3):480-4. · 5.21 Impact Factor
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    ABSTRACT: Quantitative determination of the allele frequency of single-nucleotide polymorphism (SNP) in pooled DNA samples is a promising approach to clarify the relationships between SNPs and diseases. Here, we present such a simple, accurate, and inexpensive method for quantitative determining the allele frequency in pooled DNA samples. Three steps of DNA pooling, PCR amplification and sequencing are involved in this assay. Although direct determination of the allele frequency from the two allele-specific fluorescence intensities is possible, correction for differential response of alleles is important. We explored the effect of differential response of alleles on test statistics and provide a solution to this problem based on heterozygous fluorescence intensities. We demonstrate the accuracy and reliability of this assay on pooled DNA samples with pre-determined allele frequencies from 7.1% to 53.9%. The accuracy of allele frequency measurements is high, with a correlation coefficient of r² = 0.997 between measured and known frequencies. We believe that by providing a means for SNP genotyping up to hundreds of samples simultaneously, inexpensively, and reproducibly, this method is a powerful strategy for detecting meaningful polymorphic differences in candidate gene association studies.
    Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 03/2011; 879(7-8):527-32. · 2.78 Impact Factor
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    ABSTRACT: The CD40 signaling pathway plays a key role in tumor cell proliferation, differentiation, and apoptosis. Gastric cancer usually possesses a higher level of CD40 expression than normal tissue. We evaluated inhibition of soluble CD40 ligand (sCD40L) in apoptosis induction of gastric cancer cells. Gastric cancer cells (AGS and BGC-823) were incubated with sCD40L. Cell viability and cell cycle were determined by methyl-tetrazolium (MTT) assay and flow cytometry, respectively. The results showed that sCD40L hindered cell growth, arrested cells at G0/G1 phase and induced apoptosis. In conclusion, sCD40L suppress growth of gastric cancer cells through apoptosis induction and cell cycle quiescence. This work will provided a new approach to gene therapy of gastric cancer.
    Molecular Biology Reports 03/2011; 38(8):5459-64. · 2.51 Impact Factor
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    ABSTRACT: A gastric cancer cell line, AGS has high-level expression of CD40. CD40 is present on the surfaces of a large variety of cells, including B cells, endothelial cells, dendritic cells and some carcinoma cells, and delivers signals regulating diverse cellular responses, such as proliferation, differentiation, growth suppression, cell death. In this research, the expression of CD40 and CD40 transcription in gastric cancer cell lines (AGS, BGC-823, HGC-27, SGC-7901) was investigated by semi-quantitative RT-PCR. Cancer cell proliferation inhibitory assay was also performed using different concentrations of anti-CD40 monoclonal antibody (mAb), 5C11. Results indicated that treatment of 5C11 alone and combined 5C11, IFN-alpha and 5-FU both significantly inhibited cancer cell proliferation. The synergistic effects of combined 5C11, IFN-alpha and 5-FU on growth inhibition of human gastric cancer AGS cells could be observed. These results suggest that humanized anti-CD40 monoclonal antibody (mAb), 5C11 can be used as a valuable reagent for clinical application.
    Immunology letters 02/2010; 131(2):120-5. · 2.91 Impact Factor
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    ABSTRACT: In this study, Astragalus polysaccharides was analysed using high performance liquid chromatography (HPLC) and Fourier transform infrared (FT-IR). Compared with retention time of six standard compounds in HPLC analyses, we found that the polysaccharides were composed of glucose. The absorption bands at 1639 and 1538 cm−1 were attributed to the stretching vibration of the C–O bond of carboxyl group. The absorption bands at 1035 and 956 cm−1 suggested that the extract contained pyrene monomer in its structure. The antioxidant activity of Astragalus polysaccharides was evaluated by various antioxidant assays. Astragalus polysaccharides showed strong antioxidant activity in all the tested methods. The in vitro antioxidant capacity of Astragalus polysaccharides was significantly correlated with its content. In addition to the antioxidant activity of the polysaccharides, it still showed strong antitumour activity. Then, CD40 gene siRNA plasmid (Psilencer1. 0-U6-P I3K-siRNA) was constructed. Antitumour activity of siRNA was evaluated. Among these samples, Astragalus polysaccharides exhibited more antitumour potency than siRNA. Mechanism of antitumour activity of Astragalus polysaccharides and siRNA may be explained by decreasing CD40 in cells.
    Carbohydrate Polymers. 01/2010;
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    ABSTRACT: The chemical composition of essential oil from magnolia collected in Suzhou city, China, was studied using gas chromatography-mass spectroscopy. Fifty-four volatile compounds were identified in the essential oil, with eucalyptol (21.55%), β-Pinene(11.07%), D-Limonene (8.93%), 1R-α-Pinene (7.86%), Bicyclo[2.2.1]heptan-2-one, 1,7,7-trimethyl-, (1R)-(6.46%), Cyclohexene, 4-methylene-1-(1-methylethyl)- (5.30%), 1,4-Cyclohexadiene, 1-methyl-4-(1-methylethyl)- (5.10%), camphene (4.72%), being the major chemical constituents. In an in vivo approach, the protective effect of the essential oil against oxidative stress in gastric cancer (GC) mice was also investigated in terms of superoxide dismutase, CAT, and glutathione peroxidase activities. Our results indicate that the essential oil has an effective capability to enhance serum antioxidant enzyme activities and sCD40L, and decrease phosphorylate-activate Akt protein levels in a dose-dependent manner in gastric cencer mice. Overall, essential oil from magnolia appears to be an effective antioxidant and is quite suitable for further biological evaluation. KeywordsGas chromatography-mass spectroscopy-Essential oil-Magnolia-Antioxidant-Gastric cencer-sCD40L-Phosphorylate-activate Akt protein
    Medicinal Chemistry Research 01/2010; 19(9):1203-1209. · 1.61 Impact Factor
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    ABSTRACT: To study expression of CD40 and CD40L in gastric cancer tissue we assessed gastric cancer patients admitted to the Department of Gastroenterology of The First Affiliated Hospital of Soochow University and control subjects. Gastric cancer and normal (from around tumours) tissue samples were obtained from patients. Venous blood samples (gastric cancer and ulcer groups) were drawn on the morning of the day before surgery for the measurement of peripheral sCD40L. The expression of CD40 in gastric carcinoma specimens was examined immuno-histochemically. The clinicopathological factors, including age, sex, tumor size, gross appearance, degree of cellular differentiation, histological classification, depth of tumor invasion, lymph node metastasis, peritoneal dissemination, and TNM stage were analyzed according to the different expression of CD40. The results indicated a high CD40 expression in gastric cancer tissues. This positive expression of CD40 revealed a significant (P < 0.05) correlation with lymphatic metastasis and tumor TNM stage in gastric cancer patients. It is concluded that higher CD40 expression existed in expanding type tumors and could play an important role in clinical diagnosis of gastric cancer patients.
    International Journal of Molecular Sciences 09/2009; 10(9):3900-17. · 2.46 Impact Factor
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    ABSTRACT: OBJECTIVE; To investigate the expression of CD40 in gastric cancer and explore the correlation of CD40 with the outcome of patients. The expression of CD40 was examined by immunohistochemistry in human gastric cancer tissues (73 cases) and adjacent normal gastric tissues (51 cases). CD40 mRNA was detected by RT-PCR in 11 gastric cancer and 11 adjacent normal gastric tissues. The patients were followed up for 60 months. The correlation of CD40 expression and the overall survival time were evaluated by Kaplan-Meier chart and Log-Rank test. Cox model was used for multivariate analysis. The immunoreactivity of CD40 increased in 35/73 gastric cancer tissues (47.9%) as compared to the tumor-free tissues (P = 0.0063). CD40 mRNA were detected in 72.7% (8/11) gastric cancer tissues, while not in all tumor-free tissues. After a 5-year follow-up, the survival rate was 57.9% (22/38) in CD40-negative patients, and 5.7% (2/35) in CD40-positive patients (P = 0.0022). The median survival time of patients with negative, positive, and strong CD40 expression was 56, 29 and 11 months respectively (P = 0.0085). Cox regression analysis suggested that CD40, lymph node metastasis and TNM stage were independent factors affecting the prognosis of gastric cancer patients. Normal and malignant gastric tissues are found to have its own unique CD40 expression patterns. CD40 may play a role in metastatic spread of gastric cancer and its expression may be used as an important survival predictor in human gastric cancer.
    Zhonghua yi xue za zhi 08/2009; 89(30):2124-8.
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    ABSTRACT: One major polysaccharide fractions, glucose, were isolated from the polysaccharides extract of Astragalus (AP), a valuable traditional Chinese medicine, using thin-layer chromatography (TLC) and Sephadex G-100 chromatography. HPLC and IR methods were used for a qualitative and quantitative determination of from polysaccharides of Astragalus. The HPLC method was validated for linearity, precision and accuracy. The results indicated that polysaccharides of Astragalus is an α-(1 → 4)-d-glucan with α-(1 → 6)-linked branches attached to the O-6 of branch points. Bioactivity tests showed that polysaccharides of Astragalus is active for spleen lymphocytes proliferation. The polysaccharides also presented anti-inflammatory activities. These data together suggest that polysaccharides of Astragalus presents significant immune modulating activity, thus supporting the popular use of the polysaccharides in the treatment of gastric cancer diseases.
    Carbohydrate Polymers. 01/2009;