Structure elucidation and anticancer activity of a heteropolysaccharide from white tea.

White tea, one of the six traditional teas in China, is made only through natural withering and low-temperature drying processes. It demonstrates diverse pharmacological and health-promoting effects, including antioxidant, antiviral, anticancer, and hypolipidemic activities. Despite the significance of polysaccharides in white tea leaves, their fine structure and physiological functions remain unexplored. In this study, the polysaccharide fragment WTP-80a with anticancer activity was isolated and purified from white tea through water extraction, alcohol precipitation, DEAE-52 ion exchange column chromatography, and sephacryl S-200 dextran gel column chromatography. WTP-80a exhibited a molecular weight of 1.14 × 105 Da and consisted of galactose (Gal), arabinose (Ara), rhamnose (Rha), and glucuronic acid (Glc-UA). The main chain skeleton of WTP-80a contained 3,6)-ß-Galp-(1→, 3)-α-Galp-(1→, 5)-α-Araf-(1 â†’ and 3)-α-Glcp-UA-(1→. Branch chains included α-Araf-(1 â†’ and ß-Rhap-(1 â†’ connected to the C3 and C6 positions of →3,6)-ß-Galp-(1→, respectively. In vitro anticancer experiments revealed that WTP-80a effectively hindered the proliferation, colony formation, migration, and invasion of B16F10 cells. Additionally, it induced apoptosis in B16F10 cells by blocking the G2/M phase, increasing active oxygen content, and reducing mitochondrial membrane potential. These findings provide a solid theoretical foundation for the application of white tea polysaccharides as anticancer products.

Bioinformatics analysis for the identification of Sprouty-related EVH1 domain-containing protein 3 expression and its clinical significance in thyroid carcinoma.

The poorly differentiated thyroid carcinoma (THCA) subtype is associated with an aggressive disease course, a less favorable overall prognosis, and an increased risk of distant organ metastasis. In this study, our objective was to explore the potential utility of the Sprouty-related EVH1 domain-containing protein 3 (SPRED3) as a biomarker for early diagnosis and prognosis in THCA patients. The differentially expressed prognostic-related genes associated with THCA were identified by querying The Cancer Genome Atlas (TCGA) database. The difference in the expression of the SPRED3 gene between thyroid carcinoma (THCA) tissues and normal tissues was analyzed using data from The Cancer Genome Atlas (TCGA) and further validated through immunohistochemistry. Univariate and multivariate Cox regression models were used, along with clinical information from THCA patients, to analyze the prognostic value of the SPRED3 gene in THCA patients. Functional enrichment analysis was subsequently performed to elucidate the molecular mechanisms underlying the regulatory effects of the SPRED3 gene on thyroid carcinoma. Additionally, we calculated the percentage of infiltrating immune cells in THCA patients and evaluated their correlation with SPRED3 gene expression. Compared with those in noncancerous thyroid tissue, the gene and protein expression levels of SPRED3 were found to be elevated in thyroid carcinoma tissues. Furthermore, the expression of SPRED3 in thyroid carcinoma exhibited significant correlations with tumor location, histological grade, pathological stage, and tumor node metastasis classification (TNM) stage. Univariate and multivariate Cox proportional hazards (Cox) regression analyses demonstrated that SPRED3 could serve as an independent prognostic factor for predicting the overall survival of THCA patients. The results of functional enrichment analysis suggested the potential involvement of SPRED3 in the regulation of extracellular matrix organization, epidermal development, signaling receptor activator activity, skin development, receptor ligand activity, glycosaminoglycan binding, neuroactive ligand‒receptor interaction, the IL-17 signaling pathway, and the PI3K-Akt signaling pathway. Additionally, there were significant correlations between the expression level of the SPRED3 gene and the infiltration of various immune cells (eosinophils, central memory T cells, neutrophils, macrophages, and NK cells) within the thyroid tumor microenvironment. SPRED3 can be used as a prognostic biomarker in patients with THCA could potentially be therapeutic target for THCA.

NINJ2 deficiency inhibits preadipocyte differentiation and promotes insulin resistance through regulating insulin signaling.

OBJECTIVE: Genetic variants in ninjurin-2 (NINJ2; nerve injury-induced protein 2) confer risk of ischemic strokes and coronary artery disease as well as endothelial activation and inflammation. However, little is known about NINJ2's in vivo functions and underlying mechanisms. METHODS: The phenotypes of NINJ2 knockout mice were analyzed, and mechanisms of NINJ2 that regulate body weight, insulin resistance, and glucose homeostasis and lipogenesis were investigated in vivo and in vitro. RESULTS: This study found that mice lacking NINJ2 showed impaired adipogenesis, increased insulin resistance, and abnormal glucose homeostasis, all of which are risk factors for strokes and coronary artery disease. Mechanistically, NINJ2 directly interacts with insulin receptor/insulin-like growth factor 1 receptor (INSR/IGF1R), and NINJ2 knockdown can block insulin-induced mitotic clonal expansion during preadipocyte differentiation by inhibiting protein kinase B/extracellular signal-regulated kinase (AKT/ERK) signaling and by decreasing the expression of key adipocyte transcriptional regulators CCAAT/enhancer-binding protein ß (C/EBP-ß), C/EBP-α, and peroxisome proliferator-activated receptor γ (PPAR-γ). Furthermore, the interaction between NINJ2 and INSR/IGF1R is needed for maintaining insulin sensitivity in adipocytes and muscle via AKT and glucose transporter type 4. Notably, adenovirus-mediated NINJ2 overexpression can ameliorate diet-induced insulin resistance in mice. CONCLUSIONS: In conclusion, these findings reveal NINJ2 as an important new facilitator of insulin receptors, and the authors propose a unique regulatory mechanism between insulin signaling, adipogenesis, and insulin resistance.

NPF transporters in synaptic-like vesicles control delivery of iron and copper to seeds.

Accumulation of iron in seeds is essential for both plant reproduction and human nutrition. Transport of iron to seeds requires the chelator nicotianamine (NA) to prevent its precipitation in the plant vascular tissues. However, how NA is transported to the apoplast for forming metal-NA complexes remains unknown. Here, we report that two members of the nitrate/peptide transporter family, NAET1 and NAET2, function as NA transporters required for translocation of both iron and copper to seeds. We show that NAET1 and NAET2 are predominantly expressed in the shoot and root vascular tissues and mediate secretion of NA out of the cells in resembling the release of neurotransmitters from animal synaptic vesicles. These findings reveal an unusual mechanism of transmembrane transport in plants and uncover a fundamental aspect of plant nutrition that has implications for improving food nutrition and human health.

Human Leukocyte Antigen Profile Predicts Severity of Autoimmune Liver Disease in Children of European Ancestry.

BACKGROUND AND AIMS: Genetic predisposition to autoimmune hepatitis (AIH) in adults is associated with possession of human leukocyte antigen (HLA) class I (A*01, B*08) and class II (DRB1*03, -04, -07, or -13) alleles, depending on geographic region. Juvenile autoimmune liver disease (AILD) comprises AIH-1, AIH-2, and autoimmune sclerosing cholangitis (ASC), which are phenotypically different from their adult counterparts. We aimed to define the relationship between HLA profile and disease course, severity, and outcome in juvenile AILD. APPROACH AND RESULTS: We studied 236 children of European ancestry (152 female [64%], median age 11.15 years, range 0.8-17), including 100 with AIH-1, 59 with AIH-2, and 77 with ASC. The follow-up period was from 1977 to June 2019 (median 14.5 years). Class I and II HLA genotyping was performed using PCR/sequence-specific primers. HLA B*08, -DRB1*03, and the A1-B8-DR3 haplotype impart predisposition to all three forms of AILD. Homozygosity for DRB1*03 represented the strongest risk factor (8.8). HLA DRB1*04, which independently confers susceptibility to AIH in adults, was infrequent in AIH-1 and ASC, suggesting protection; and DRB1*15 (DR15) was protective against all forms of AILD. Distinct HLA class II alleles predispose to the different subgroups of juvenile AILD: DRB1*03 to AIH-1, DRB1*13 to ASC, and DRB1*07 to AIH-2. Possession of homozygous DRB1*03 or of DRB1*13 is associated with fibrosis at disease onset, and possession of these two genes in addition to DRB1*07 is associated with a more severe disease in all three subgroups. CONCLUSIONS: Unique HLA profiles are seen in each subgroup of juvenile AILD. HLA genotype might be useful in predicting responsiveness to immunosuppressive treatment and course.

Comparison of conbercept and ranibizumab combined mitomycin C-augmented trabeculectomy for neovascular glaucoma.

PURPOSE: To compare the efficacy and security of conbercept and ranibizumab combined with trabeculectomy and panretinal photocoagulation for neovascular glaucoma (NVG). METHODS: One hundred and sixty patients with NVG were randomly divided into a conbercept group comprised of 80 patients and a ranibizumab group comprised of 80 patients. The postoperative and preoperative visual acuities, intraocular pressures frequency of anti-glaucoma medications, and surgical complications were recorded. The expressions of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (FLT-1), and placenta-like growth factor (PLGF) in the aqueous humor were determined using an enzyme-linked immunosorbent assay. Examining the fundus and obtaining photographs used indirect ophthalmoscopy. Kaplan-Meier and log-rank analyses estimated the success rates. RESULTS: All patient follow-up periods were at 1 year. The differences observed in IOP and the frequencies of anti-glaucoma medications at various follow-up time points were not statistically significant (all P > 0.05). The differences observed in both the group visual acuities at various follow-up time points were not statistically significant (P > 0.05). Rates of surgery complications were 18.75% and 25.00% in the conbercept group and ranibizumab group, respectively. The expressions of VEGF, FLT-1, and PLGF significantly decreased (all P < 0.05). The recurrence percentages were 30.00% and 36.25% after conbercept and ranibizumab treatment, respectively. CONCLUSION: The conbercept effects were similar with that of ranibizumab. Intravitreal injection of conbercept was effective for NVG treatment, which provides a new therapeutic drug for NVG treatment.

C-terminal of E1A binding protein 2 promotes the malignancy of osteosarcoma cells via JAK1/Stat3 signaling.

Previous studies have demonstrated that the C-terminal of E1A binding proteins (CtBPs) influences tumorigenesis by participating in cell signal transduction in various human malignancies. However, the detailed expression patterns of CtBP isoforms in human osteosarcoma (OS) and the molecular mechanisms of CtBP involvement in tumor cell phenotypes requires further investigation. In the present study, the expression patterns of CtBP2 in OS cells and tissues were explored by immunohistochemistry. Fetal osteoblast cells were transfected with a eukaryotic expression plasmid to overexpress CtBP2, and the endogenous CtBP2 in OS cells was silenced via a short hairpin RNA. These transfections were validated and the phosphorylation levels of the JAK1/Stat3 signaling pathway were explored via western blotting. Furthermore, the malignant phenotype of OS cells was evaluated via a Cell Counting Kit-8 assay, cell colony formation assay, cell migration assay and scratch wound healing assay. The results revealed that the expression of CtBP2, but not CtBP1, was upregulated in OS tissue samples and the elevated expression level of CtBP2 was notably associated with distant metastasis. CtBP2 was demonstrated to modulate cell migration and invasion via JAK1/Stat3 signaling pathway in fetal osteoblast cells. In addition, genetic silencing of CtBP2 expression in OS cells notably reduced cell migration abilities and the phosphorylation of the JAK1/Stat3 pathway. In summary, the present studies revealed that the loss of CtBP2 constrained distant metastasis through the JAK1/Stat3 pathway in OS, suggesting that targeting CtBP2 may be a practical anti-tumor approach to prevent OS tumor progression.

Major vault protein plays important roles in viral infection.

Viral replication and related protein expression inside the host cells, and host antiviral immune responses can lead to the occurrence of diverse diseases. With the outbreak of viral infection, a large number of newly diagnosed and died patients infected with various viruses are still reported every year. Viral infection has already been one of the major global public health issues and lead to huge economic and social burdens. Studying of viral pathogenesis is a very important way to find methods for prevention, diagnosis, and cure of viral infection; more evidence has confirmed that major vault protein (MVP) is closely associated with viral infection and pathogenesis, and this review is intended to provide a broad relationship between viruses and MVP to stimulate the interest of related researchers.

Inflammatory cytokines in highly myopic eyes.

Currently, myopic retinopathy is the most common irreversible blinding disease but its pathophysiology is not completely clear. A cross-sectional, observational study was conducted in a single center to analyze aqueous samples from highly myopic eyes (axial length >25 mm, n = 92) and ametropic or mild myopic eyes (n = 88) for inflammatory cytokines. Vascular endothelial growth factor (VEGF), Interleukin 6 (IL-6), and matrix metalloproteinase-2 (MMP-2) were measured using an enzyme-linked immunosorbent assay. IL-6 and MMP-2 were significantly higher in the highly myopic eyes than in the non-high myopic eyes (IL-6: 11.90 vs. 4.38 pg/mL, p < 0.0001; MMP-2: 13.10 vs. 8.82 ng/mL, p = 0.0003) while adjusting for age, gender, and intraocular pressure. There was a significant positive association between levels of IL-6 and MMP-2 in aqueous humor and the axial lengths of the eye globes (IL-6, ß = 0.065, p < 0.0001, n = 134; MMP-2, ß = 0.097, p < 0.0001, n = 131). Conversely, VEGF in aqueous humor was significantly lower in the highly myopic eyes than in the non-high myopic eyes (45.56 vs. 96.90 pg/mL, p < 0.0001, n = 153) while age, gender, and intraocular pressure were adjusted. The results suggest that low-grade intraocular inflammation may play an important role in the development and progression of high myopia and myopic retinopathy.

Inhibition of migration and invasion by berberine via inactivation of PI3K/Akt and p38 in human retinoblastoma cell line.

BACKGROUND: As a clinically important natural isoquinoline alkaloid, berberine has been reported to possess various pharmacological effects. OBJECTIVES: This study was aimed to investigate the effect of berberine on cell migration and invasion in human retinoblastoma (Rb) cells. MATERIAL AND METHODS: The cytotoxicity of berberine was estimated by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay. After being stimulated with berberine under various concentrations, the cell migration and invasion were evaluated by transwell assay. Then, the expression levels of epithelial-mesenchymal transition (EMT) markers were determined by quantitative reverse transcription PCR (qRT-PCR) and western blot analysis. Furthermore, the phosphorylation levels of protein kinase B (Akt) and p38 were detected by western blot analysis. Finally, the effect of phosphatidylinositol-3-kinase (PI3K) and p38 inhibitors on cell migration and invasion was estimated by transwell assay. Untreated cells acted as control for all the experiments. RESULTS: The concentrations of berberine for further studies were controlled in a range of 0 to 100 µM. The cell migration and invasion were both suppressed by berberine in a dose-dependent manner compared to the control (p < 0.05 or p < 0.001). Berberine remarkably down-regulated expression of E-cadherin and up-regulated expression of vimentin and α-SMA compared to the control (p < 0.01 or p < 0.001). Furthermore, the phosphorylation levels of Akt and p38 were both down-regulated by berberine in comparison to the control. Furthermore, the addition of berberine accompanied by LY294002 or SB203580 significantly suppressed cell migration and invasion compared to the addition of berberine alone (p < 0.05). CONCLUSIONS: Berberine suppressed cell migration and invasion via inactivation of PI3K/Akt and p38.

A preliminary comparative clinical study of vertebroplasty with multineedle or single-needle interstitial implantation of (125)I seeds in the treatment of osteolytic metastatic vertebral tumors.

OBJECT: Percutaneous vertebroplasty (PVP) combined with brachytherapy using the interstitial implantation of (125)I seeds has previously yielded encouraging clinical results in the treatment of metastatic vertebral tumors. However, the bone cement injection volume is very small due to the osteolytic damage to the metastatic vertebrae, and the ideal spatial distribution of the (125)I seeds is difficult to achieve. In the current study, the authors present a clinical method for puncture needle insertion to achieve a greater bone cement injection volume and a more ideal spatial distribution of the (125)I seeds. METHODS: Twenty-nine patients with osteolytic metastatic vertebral tumors were divided into 2 groups and were treated with either PVP combined with multineedle interstitial implantation of (125)I seeds, or PVP combined with single-needle interstitial implantation of (125)I seeds. Clinical efficacy was evaluated according to a visual analog scale (VAS) of pain, the Karnofsky Performance Scale (KPS), and the Response Evaluation Criteria In Solid Tumors (RECIST). RESULTS: Back pain was significantly alleviated in all patients after surgery. Compared with the preoperative scores, the VAS scores were significantly decreased in both groups at 1 week and 3 months postoperatively (p < 0.05), but there were no significant intergroup differences (p > 0.05). The postoperative quality of life was improved in both groups; the KPS scores increased significantly compared with the preoperative scores (p < 0.05), and the postoperative KPS scores were significantly different between the 2 groups (p < 0.05). No intergroup differences were observed in pain alleviation, but the bone cement injection volume was significantly greater in the multineedle group than in the single-needle group (p < 0.05). The clinical benefit rate and disease control rate at 3 months after the operation were both significantly better for the multineedle group (p < 0.05). CONCLUSIONS: The outcomes of PVP combined with multineedle interstitial implantation of (125)I seeds in patients with osteolytic metastatic vertebral tumors appeared to be better than the outcomes of PVP combined with single-needle interstitial implantation of (125)I seeds. These better outcomes may be the result of the greater bone cement injection volume and the more ideal spatial distribution of the (125)I seeds.

Autoantibody and human leukocyte antigen profiles in children with autoimmune liver disease and their first-degree relatives.

OBJECTIVE: Familial clustering of juvenile autoimmune liver disease (AILD), including autoimmune hepatitis and autoimmune sclerosing cholangitis (ASC), is rare, despite a high prevalence of autoimmune disorders in AILD families. METHODS: To investigate this discrepancy, we measured autoantibodies diagnostic for AILD, anti-nuclear, anti-smooth muscle, anti-liver kidney microsomal type 1, anti-liver cytosol type 1, and anti-soluble liver antigen antibodies, and human leukocyte antigen profiles in 31 patients and 65 of their first-degree relatives (FDR). The autoantibody profile was compared with that of 42 healthy subjects (HS). RESULTS: Autoantibodies were detected in 71% (22/31) patients. Anti-nuclear antibody or anti-smooth muscle antibody were present in 4/65 FDR (6.2%). HS were negative for all autoantibodies. The frequencies of homozygous HLA DRB1*0301 (DR3) genes and haplotype A1-B8-DR3 were higher in the patients (25% and 43%) than in FDR (9% and 27%) and HS (0% and 16%). The frequencies of disease-protective genes DR4 and/or DR15 were lower in the patients (25%) than in FDR (42%) and HS (42%). Only 1 family contained 2 patients with AILD, 1 with ASC and 1 with primary sclerosing cholangitis. Both patients possessed A1-B8-DR3 genes, the ASC being homozygous and the primary sclerosing cholangitis heterozygous. Six FDR had nonhepatic autoimmune disorders, none being autoantibody positive. CONCLUSIONS: Homozygosity for DR3 plays a major role in the predisposition to juvenile AILD. Diagnostic autoantibodies for AILD are rare among patients' FDR and not linked to clinical manifestation of AILD.

Clinical assessment of reformed lumbar microdiscectomy.

The aim of this study was to evaluate the clinical outcomes of the reformed lumbar microdiscectomy preserving more ligamentum flavum than the traditional microdiscectomy does. A prospective, randomized, controlled clinical study was conducted. Patients with unilateral lumbar disc herniation were randomly divided into two groups. The control group underwent traditional lumbar microdiscectomy, and the test group patients underwent the same procedure but with a curved incision of the lumbodorsal fascia and with more preservation of the ligamentum flavum. Visual analogue scale (VAS) scores and Oswestry scale scores were used to appraise the outcomes. Both groups' clinical parameters were significantly improved after the operation. The VAS scores in the test group showed a less intensity than that in the control group at 3 days, 12 weeks after the operation (P < 0.05), while at 1 year, showed no significant difference. Both groups' postoperative leg pain was significantly relieved (P < 0.05). The VAS scores for leg pain had no significant difference between the two groups at any testing time point after the surgery (P > 0.05). The Oswestry scale scores showed a better lumbar function state in the test group than that in the control group at 12 weeks and 1 year after the operation (P < 0.05). In both groups, there was no patient who had a lumbar disc reherniation. Preserving more ligamentum flavum is helpful to improve the clinical outcomes, and this improvement maybe resulted from the prevention of the fibrosis-related complication and the stability of the spine.

Antitumor and Immunomodulatory Effects of Polysaccharides from Broken-Spore of Ganoderma lucidum.

The antitumor and immunomodulatory activity of broken-spore of Ganodermalucidum polysaccharides (Gl-BSP) were investigated in vivo and in vitro. It was showed that Gl-BSP (50, 100, and 200 mg kg(-1)) exhibited antitumor effect against Sarcoma 180 (S180) in BALB/c mice. The Gl-BSP was not cytotoxicity in S180 cells and PG cells (human lung carcinoma cell) in vitro. However, serum from Gl-BSP-treated S180-bearing mice significantly inhibited S180 and PG cells proliferation in vitro. Moreover, Gl-BSP promoted the splenic lymphocyte proliferation induced by Con A or LPS, enhanced nature killer cell (NK cell) cytotoxic activity, augmented the percentage of neutral red phagocytosis by macrophages, and increased the percentage of the CD4(+) or CD8(+) subset in S180-bearing mice. The serum level of IFN-γ, TNF-α, and nitric oxide was increased by Gl-BSP. Gl-BSP also showed immunomodulatory activities in tumor-bearing mice. Furthermore, neutralization with anti-TNF-α and/or anti-IFN-γ significantly diminished growth inhibition induced by Gl-BSP-treated serum of S180-bearing mice in S180 or PG cells. These observations suggest that the antitumor activity of Gl-BSP may be mainly related to the activation of the immune response of the host organism by the stimulation of NK cells, T cells, and macrophages.

Human CD4+CD25(high)CD127 (low/neg) regulatory T cells.

CD4+CD25(high)CD127(low/neg) regulatory T cells (Tregs) play a critical role in the maintenance of peripheral tolerance and in controlling the development of autoimmune diseases. A combination of surface and intracellular markers, namely, CD25, CD39/CD73, CD62L, CD45RO, CD127, glucocorticoid-induced tumor necrosis factor receptor (GITR), CTLA-4, and the forkhead/winged helix transcription factor (FOXP3), has been used to characterize Tregs. Tregs suppress T effector responses mainly in a direct cell-cell contact manner. However, other mechanisms independent from this manner cannot be excluded entirely. It has been shown that Tregs can undergo limited expansion in vitro after the stimulation of TCR in the presence of exogenous cytokines, e.g., IL-2. Expanded Tregs retain their suppression function. Human Tregs have demonstrated their great potential to be used as a therapeutic intervention in preventing graft rejection and treating autoimmune diseases. In this chapter, we have given a review on how to characterize, isolate, expand Tregs and assess their suppressive functions.

[The pharmacological study of Ganoderma spores and their active components].

Recently, the pharmacological study of Ganoderma spores and active components has become a focus of attention in the world. The present review is based on the auctorial research on Ganoderma spores. It involves pharmacological effects of Ganoderma spores and its active components, including immunomodulating effect, antitumor activity and its mechanism in vivo and in vitro, liver-protective effect, gastric ulcer preventing effect, serum glucose and blood fat depressing effects, anti-hypoxia and scavenging free redical, etc. The possible problems and their solutions in this research area are also discussed.

[Comparison of the immunomodulatory effects of spore polysaccharides and broken spore polysaccharides isolated from Ganoderma lucidum on murine splenic lymphocytes and peritoneal macrophages in vitro].

OBJECTIVE: To compare the immunomodulatory effects of spore polysaccharides (Gl-SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum(Leyss et Fr.) Karst. on murine splenic lymphocytes and peritoneal macrophages in vitro. METHODS: Mixed lymphocyte culture reaction (MLR), lymphocyte proliferation in the presence or absence of mitogen, and the cytotoxic activity of splenic natural killer (NK) cells were detected with MTT assay in vitro. The percentage of phagocytosis of neutral red (NR) by mouse peritoneal macrophages was detected by colorimetric assay. Splenic T-lymphocyte subpopulations were measured with flow cytometry(FCM). IL-2, IFN-gamma and TNF-alpha in the culture supernatants were detected by ELISA and biological assay. Nitric oxide (NO) production was examined by Griess reaction. RESULTS: At the concentration range of 0.2-12.8 mg/L, Gl-SP and Gl-BSP were shown to increase lymphocyte proliferation in the presence or absence of mitogen, enhance NK cytotoxic activity, augment the production of TNF-alpha and NO in Gl-SP- or Gl-BSP-activated macrophages, as well the percentage of phagocytosis of NR by macrophages in vitro. Both Gl-SP and Gl-BSP could promote MLR, however, at the dose of 12.8 mg/L, Gl-BSP showed higher activity than Gl-SP in the proliferation of lymphocytes. These two kinds of polysaccharide could significantly increase the secretion of IL-2 and IFN-gamma in doublejway MLR at the concentrations of 0.2-12.8 mg/L, but Gl-BSP had stronger effects than Gl-SP at the same concentrations. Both Gl-SP and Gl-BSP could increase the ratio of T-lymphocyte subpopulations in double-way MLR. At the concentrations of 0.2-12.8 mg/L or 3.2-12.8 mg/L, Gl-BSP demonstrated more significant activity in increasing the percentage of the CD4(+) or CD8(+) subset than Gl-SP. At the concentrations of 0.2-0.8 mg/L, the ratio of the CD4(+) and CD8(+) subset in the Gl-BSP treated group was higher than that of the Gl-SP treated group. CONCLUSION: Gl-SP and Gl-BSP have similar immunomodulatory effects in vitro, as though the immunomodulatory effects of Gl-BSP are stronger than that of Gl-SP.