Renal protective effect of Ganoderma lucidum / 中国药理学与毒理学杂志

OBJECTIVE Non-alcoholic fatty liver disease(NAFLD) encompasses a series of patho-logic changes ranging from steatosis to steatohepatitis,which may progress to cirrhosis and hepatocel-lular carcinoma.The purpose of this study was to determine whether Ganoderma lucidum polysaccha-ride peptide (GLPP) has therapeutic effect on NAFLD. METHODS ob/ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD. Key metabolic path-ways and enzymes were identified by metabolomics combining with KEGG and PIUmet analyses and key enzymes were detected by Western blotting. Hepatosteatosis models of HepG2 cells and primary hepatocytes were used to further confirm the therapeutic effect of GLPP on NAFLD. RESULTS GLPP administrated for a month alleviated hepatosteatosis, dyslipidemia, liver dysfunction and liver insulin resistance. Pathways of glycerophospholipid metabolism, fatty acid metabolism and primary bile acid biosynthesis were involved in the therapeutic effect of GLPP on NAFLD. Detection of key enzymes revealed that GLPP reversed low expression of CYP7A1,CYP8B1,FXR,SHP and high expression of FGFR4 in ob/ob mice and ApoC3 mice. Besides, GLPP inhibited fatty acid synthesis by reducing the expression of SREBP1c, FAS and ACC via a FXR-SHP dependent mechanism. Additionally, GLPP reduced the accumulation of lipid droplets and the content of TG in HepG2 cells and primary hepato-cytes induced by oleic acid and palmitic acid. CONCLUSION GLPP significantly improves NAFLD via regulating bile acid synthesis dependent on FXR-SHP/FGF pathway, which finally inhibits fatty acid synthesis,indicating that GLPP might be developed as a therapeutic drug for NAFLD.

Efficacy of sublingual immunotherapy with dermatophagoides farinae drops in patients with allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of sublingual immunotherapy (SLIT) with dermatophagoides farinae drops in patients with allergic rhinitis (AR).</p><p><b>METHODS</b>Two hundred and six patients with AR aged from 4 to 60 years were included in this study. Among them, 123 patients completed more than one year treatment, the number of patients completed one year, one year and a half, two years were 61, 41, 21 cases. Rhinitis symptom scores and medication scores of the three groups before and after treatment were compared. And rhinitis symptom scores and medication scores of patients with one year drug withdrawal after one and two years treatment were aslo compared.</p><p><b>RESULTS</b>After SLIT one year, one year and a half, two years treatment, the symptoms in these patients were significantly improved compared with before. The symptom scores (x±s) were reduced from 6.00±2.27, 7.39±1.99 and 6.29±2.14 to 2.95±1.82, 3.28±2.58, 2.48±1.99. The differences were statistically significant (t value was 8.19, 10.29, and 5.97, all P<0.01). The proportion of patients without drug treatment of the three group were 68.9%, 73.2% and 80.9%, there was statistical significance before and after treatment in every group (value was 50.391, 43.619, 27.776, all P<0.01). Symptom improvement of sneezing, runny nose, nasal congestion, nasal itching were similar after one year, one year and a half, two years SLIT treatment, the differences were not statistically significant (F values were 1.200, 1.276 and 2.333, all P>0.05). The proportion of patients without drug treatment in the group which stopped medication one year after two year SLIT (76.2%) was higher than group stopped medication one year after one year SLIT (61.3%). There was no statistical significance (χ2=1.263, P>0.05).</p><p><b>CONCLUSION</b>The dust mite drops can relieve symptoms after one year treatment, but the proposed two years treatment is important for the consolidation of improved symptoms, especially for the effect of reducing the use of symptomatic medication.</p>

Effects of allergen and intranasal glucocorticoid on Th17 and RORgamma t in peripheral blood in patients with allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the effect of specific immunotherapy and intranasal glucocorticoid on T help 17 (Th17) cells and RORgammat in peripheral blood in patients with allergic rhinitis (AR).</p><p><b>METHODS</b>Forty patients with allergic rhinitis (group A) were divided randomly into two subgroups (group A1 and A2), and each subgroup had 20 patients. The patients in group A1 were treated with intranasal glucocorticoid (INGS) for one-year. The patients in group A2 were treated with special immunotherapy (SIT) for 4 weeks. Blood samples were respectively taken from 10 healthy individuals (group B), 20 AR patients (group A1) before and after SIT with specific standardized allergen and 20 AR patients (group A2) before and after INGS. The ratio of Th17 cells in peripheral blood monouclear cells (PBMC) were analysed by flow cytometry. The expression of RORgammat mRNA were detected by real-time polymerase chain reaction and the interleukin-23(IL-23), IL-17, IL-6 were detected by enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>The ratio of Th17 cells in PBMC and the expression of RORgammat mRNA in group A [(18.97 +/- 1.05)% and (0.604 +/- 0.027)] were respectively higher than those in group B [(15.12 +/- 1.09)% and (0.447 +/- 0.024)] and the difference reached statistical significance (t were respectively -10.056 and -17.986, each P < 0.01). The level of IL-6, IL-17 and IL-23 in group A were respectively higher than those in group B and the difference reached statistical significance (t were respectively -41.149, -17.618 and -26.824, all P < 0.01). The ratio of Th17 cells in PBMC, the expression of RORgammat mRNA, the level of IL-6, IL-17 and IL-23 before INGS did not show significant difference from those of after INGS in group A1 (t were respectively 0.298, 0.240, -1.136, 0.283 and -1.670, all P > 0.05). The ratio of Th17 cells in PBMC and the expression of RORgammat mRNA were respectively (18.99 +/- 1.14)% and (0.603 +/- 0.027) before SIT and were respectively (16.30 +/- 1.63)% and (0.429 +/- 0.023) after SIT in group A2, and the difference reached statistical significance (t were respectively 6.035 and 22.015, all P < 0.01). The level of IL-6, IL-17 and IL-23 before SIT were lower respectively than those of after SIT in group A2 and the difference reached statistical significance (t were respectively 9.235, 11.289, 7.267, all P < 0.01).</p><p><b>CONCLUSIONS</b>The ratio of Th17 cells in PBMC, the expression of RORgammat mRNA, the level of IL-6, IL-17 and IL-23 were up-regulated in patients with AR. The treatment of SIT could get the 5 items down and the treatment of INGS couldn't.</p>

Isolation, purification and structural analysis of GL-PP-3A, an active polysaccharide peptide from Ganoderma lucidum / 药学学报

GL-PP-3A, an active polysaccharide peptide, was isolated and purified from Ganoderma lucidum, and then its structure was analyzed. Crude polysaccharide peptides were extracted from Ganoderma lucidum with hot water, precipitated with ethanol and then dialyzed from Ganoderma lucidum. Subsequently GL-PP-3A was isolated and purified from the crude polysaccharide peptides by fractional precipitation and chromatography of Bio-Gel P-10 column. The repetitive unit of GL-PP-3A was analyzed by high performance gel permeation chromatography (HPGPC), monosaccharide composition and methylation analysis, 1H NMR and 13C NMR. GL-PP-3A is a heteropolysaccharide which is composed mainly of glucose (Glc), and also contains saccharide residues such as rhamnose (Rha), xylose (Xyl), mannose (Man) and galactose (Gal) and 17 kinds of amino acids. Its weight-average molecular weight (Mw) and number-average molecular weight (Mn) were 1.7 x 10(4) and 1.1 x 10(4), respectively, with the ratio of Mw/Mn ( molecular weight distribution) being of 1.49. Its backbone chain is composed of 1,6-linked beta-D-Glcp and 1,3-liked beta-D-Glcp at a ratio of 2:1. Some of 1,6-linked glucose residuals of the backbone chain are substituted at 2-0 or 3-0, and there are 1 to 3 1,6-linked beta-D-Galp or 1,3-linked alpha-D-Manp in the branched chains, the nonreducing ends of which consist mainly of beta-D-Glcp and a few Rha.

Detection of specific IgE of anti-Staphylococcus aureus enterotoxins in nasal polyps and analysis theoretically about the superantigen hypothesis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To search the evidence for the presence of superantigen of Staphylococcus aureus enterotoxin (SE) in the pathogenesis of nasal polyposis.</p><p><b>METHODS</b>In a cohort of population composed of 42 cases who belonged to three groups: nasal polyposis, simple chronic rhinosinusitis (CRS), and control group without any rhinopathy, detecting the specific IgE against SE-A and B (SEA and SEB), total IgE (TIgE), eosinophilic cationic protein (ECP) of the local mucosa by means of FRAST (UniCAP system), as well as the serum TIgE, and serum anti-SEA and SEB SIgE (only in 8 cases); meanwhile the secretion culture was performed for aerobic bacteria from the middle meatus.</p><p><b>RESULTS</b>There was no evidence to support that SE played as a superantigen in all mucosa samples (42 cases) and 8 cases serum samples out of the 42 patients. The range of TIgE in mucosa was 4.59 -70.21 kIU/2 mg tissue protein, the mean was (17.85 +/- 14.31) kIU/2 mg tissue protein; in serum the total IgE was 7.44 - 344.00 kIU/L, the mean was (88.65 +/- 80.03) kIU/L The positive culture of Staphylococcus aureus was obtained from only 3 cases from secretion of middle meatus (1 from nasal polyps, 2 from CRS). There was no significance statistically among the three groups on the tissue fluorescence value of SIgE for SE, the means of tissue TIgE and ECP.</p><p><b>CONCLUSIONS</b>No evidence was found to support the role of SE acting as a superantigen among our cases who did not have persistent asthma. It is suggested that further study and investigation is required to prove the superantigen Hypothesis in the pathogenesis of NPs.</p>

Expression of T-bet and its relation with IgE and eosinophil cationic protein in patients with allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of T-bet mRNA in peripheral blood mononuclear cells (PBMC) as well as its relations with total IgE (TIgE), eosinophil cationic protein (ECP) in serum and nasal allergic symptoms in patients with allergic rhinitis (AR).</p><p><b>METHODS</b>The allergen, TIgE and ECP in serum of patients with AR were detected by Unicap CAP system. Blood sample was taken from 8 healthy individuals and 22 patients with allergic rhinitis. PBMC was isolated by density gradient centrifugation and one part of them was cultured with 50 microg/ml mite allergen. PBMC was subjected to analysis of T-bet mRNA expression using semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The ratio of T-bet to beta-actin mRNA levels was 0.381 +/- 0.099 in patients and 0.750 +/- 0.067 in normal individuals, the difference was significantly (P <0.01). The expression intensity of T-bet mRNA had no relation to varying severity of allergic symptoms and concentration of ECP and the correlation coefficient was 0.187 and -0.165 (all P > 0.05). However, there was an inverse correlation between expression intensity of T-bet mRNA and TIgE concentration (r = -0.525, P < 0.05). Mean mRNA level (x +/- s) of T-bet expression before and after being stimulated by allergen was 0.381 +/- 0.099 and 0.365 +/- 0.104 respectively, which indicated no significant differences (P > 0.05).</p><p><b>CONCLUSIONS</b>Among allergic patients whose allergen was mite, there was a down-regulated expression of T-bet mRNA, which had no relation to ECP concentration and allergic symptoms, but was one of important links in mechanisms of imbalance of Th1/Th2 in AR. There was no effect of specific allergen on T-bet mRNA in patients with AR T-bet was one of indirect factors that affected the level of IgE.</p>

Immunopharmacological action of sinomenine, an alkaloid isolated from Sinomenium acutum, and its mechanism of action in treating rheumatoid arthritis / 药学学报

<p><b>AIM</b>To observe the effects of sinomenine on the immune functions and apoptosis of murine lymphocyte as well as on human synovial fibroblast proliferation.</p><p><b>METHODS</b>Both in vivo and in vitro tests were adopted. The lymphocyte proliferation induced by mitogens was assayed by MTT method. Spleen T lymphocyte subtypes were tested with flow cytometry. Spleen lymphocyte apoptosis was analyzed by flow cytometry and DNA ladder methods. In vitro test was adopted to observe the effects of sinomenine on the proliferation of human fibroblast of rheumatoid arthritis.</p><p><b>RESULTS</b>Sinomenine can inhibit the proliferation of mouse lymphocytes induced by ConA, LPS and anti-CD3 mAb but not PMA in vitro, and inhibit the proliferation induced by LPS and PMA in vivo. Sinomenine can reduce up-regulated CD4+/CD8+ ratio of T lymphocyte subtype in adjuvant arthritis rat. At the same concentration increased apoptosis ratio. As to human synovial fibroblast, sinomenine can significantly inhibit proliferation of human fibroblast.</p><p><b>CONCLUSION</b>Sinomenine can inhibit the immunological function and correct imbalance of CD4+/CD8+ ratio of T lymphocyte subtype. It can also increase apoptosis ratio of spleen lymphocyte. This may be the mechanism of its immunological inhibitory effect.</p>

Antioxidant effect of Ganoderma polysaccharide peptide / 药学学报

<p><b>AIM</b>To study the antioxidant effect of Ganoderma polysaccharide peptide (GLPP) and its mechanism.</p><p><b>METHODS</b>Copper was used as oxidant to induce low lipoprotein (LDL) oxidative modification, and alloxan was given i.v. to induce reactive oxygen species (ROS) injury in mice.</p><p><b>RESULTS</b>GLPP decreased oxidation of LDL and the relative electrophoretic mobility (REM) of oxidative product of LDL. After GLPP was given i.p. for 20 days, the concentration of malondialdehyde(MDA) in serum and heart of mice was decreased. The GSHpx enzyme activity was increased, while the SOD level was decreased. The catalase(CAT) levels were not significantly changed by GLPP.</p><p><b>CONCLUSION</b>GLPP showed antioxidant effect by scavenging ROS or enhancing the enzyme activity of GSHpx in vivo and in vitro.</p>

Comparison of the effects of polysaccharides from wood-cultured and bag-cultured Ganoderma lucidum on murine spleen lymphocyte proliferation in vitro / 药学学报

<p><b>AIM</b>To compare the influences of wood-cultured Ganoderma lucidum polysaccharides (Gl-PS-WC) and bag-cultured Ganoderma lucidum polysaccharides (Gl-PS-BC) on the proliferation activities of murine spleen lymphocytes in vitro, and investigate whether Gl-PS-BC can be substituted for Gl-PS-WC.</p><p><b>METHODS</b>Mixed lymphocyte culture (MLC) reaction, lymphocyte proliferation induced by concanavalin A (Con A, 1 mg.L-1) or lipopolysaccharide (LPS, 5 mg.L-1), MLC reactions inhibited by immunosuppressive drugs, cyclosporine A (CsA, 0.1 mg.L-1), mitomycin (Mit C, 0.1 mg.L-1), or antitumor drug, etoposide (VP-16, 0.1 mg.L-1), were detected in the presence or absence of Gl-PS-WC and Gl-PS-BC in the concentration range of 0.2-12.8 mg.L-1.</p><p><b>RESULTS</b>Two kinds of polysaccharides were shown to promote MLC in the range of 0.2-12.8 mg.L-1, increase lymphocyte proliferation induced by Con A or LPS and antagonize the inhibitory effects of CsA, Mit C or VP-16 on MLC. No significant difference was observed between these two kinds of polysaccharides in selected concentrations.</p><p><b>CONCLUSION</b>Gl-PS-WC and Gl-PS-BC showed similar effects on the proliferation activities of murine spleen lymphocytes in vitro.</p>

Establishment of immune insulin resistance model in the rats by i.v. injection of BCG / 药学学报

<p><b>AIM</b>To observe the stability of BCG-induced insulin resistance model.</p><p><b>METHODS</b>The glucose tolerance, serum glucose, FFA, insulin, triglycerides, cholesterol, TNF-alpha and ALT level were measured. The change of GDR was measured by euglycemic clamp in model rats after given i.v. BCG 2, 4 and 8 weeks.</p><p><b>RESULTS</b>After 2, 4 and 8 weeks, the GIR and glucose tolerance of the animals deceased significantly. After 2, 4 and 8 weeks, BCG infusion resulted in a pronounced reduction in glucose tolerance and insulin-stimulated glucose disposal rate [GDR = GDR: (29 +/- 6) vs (13 +/- 7) mg.kg-1.min-1 2 weeks; (29 +/- 6) vs (11 +/- 7) mg.kg-1.min-1 4 weeks and (23 +/- 3) vs (16 +/- 3) mg.kg-1.min-1 8 weeks, respectively, P < 0.01]. BCG infusion resulted in a pronounced increase in the weights of the liver [(6.2 +/- 0.9) vs (8.2 +/- 1.3) g, P < 0.05] and spleens [(0.51 +/- 0.11) vs (1.4 +/- 0.4) g, P < 0.01]. The histo-pathological results showed that BCG infusion resulted severe inflammation in the livers and spleens and the ratio of beta/alpha in pancreas increased. The serum levels of triglyceride, FFA and glucose were unchanged, but the level of serum TNF-alpha [543 +/- 60) vs (759 +/- 137) pg.mL-1, P < 0.05] and insulin [(31 +/- 5) vs (36 +/- 5) mu.L-1, P > 0.05] increased.</p><p><b>CONCLUSION</b>This novel model of immune insulin resistance is completely and constantly established.</p>

Effects of leflunomide and its active metabolite on the production and mRNA expression of TNF-alpha in peritoneal macrophages and synovial cells with adjuvant arthritis in rats / 药学学报

<p><b>AIM</b>To observe the effects of leflunomide (LEF), an isoxazole immunomodulatory agent and its active metabolite, A771726, on the production and mRNA expression of TNF-alpha in peritoneal macrophages and synovial cells with adjuvant arthritis rats and to further investigate the immunosuppression effects of leflunomide and its mechanisms.</p><p><b>METHODS</b>ELISA methods were used for assaying the levels of TNF-alpha. The RT-PCR methods were used for measuring the expression of TNF-alpha.</p><p><b>RESULTS</b>The production of TNF-alpha was increased in the supernatant of PM psi in adjuvant arthritis (AA) model rat. LEF (5, 10, 25 mg.kg-1, ig) was shown to inhibit the release of TNF-alpha from peritoneal macrophages induced by LPS and the inhibitory effects were in a dose-effect relevance manner. A parallel investigation of cytokine mRNA expression was undertaken using semi-quantitative reverse transcribed polymerase chain reaction (RT-PCR) to follow the kinetics of cytokine appearance in PM psi and synovial membrane tissue cells obtained from AA/normal rats treated with A771726. The results of RT-PCR from macrophages and synovial membrane tissue cells of AA rats at the peak of inflammatory phase showed that TNF-alpha mRNA expression levels were higher than those of normal rats, while the expression of TNF-alpha mRNA was reduced by treating with A771726 in vitro. On the other hand, the TNF-alpha mRNA expression showed kinetics very similar to those obtained by ELISA technique which measured protein expression.</p><p><b>CONCLUSION</b>Leflunomide and its active metabolite, A771726, was found to inhibit the production and mRNA expression of TNF-alpha in peritoneal macrophages and synovial cells with adjuvant arthritis rats model. It might be involved in the mechanisms of its anti-inflammation and immunosupression.</p>