A Highly Sensitive Electrochemical Biosensor for Detection of Adenosine Triphosphate Based on Space Block / 分析化学

In this work,a highly sensitive electrochemical biosensor for the detection of trace adenosine triphosphate (ATP) was proposed.The biosensor was based on porous anodic alumina (PAA) and SiO2 nanoparticles combining with several oligonucleotides to construct sandwich structure.It was characterized by scanning electron microscopy,fluorescence microscopy,differential pulse voltammetry and electrochemical impedance spectroscopy,which conformed to the reliability of the biosensor fabrication and the feasibility of the detection.In the presence of ATP,the sandwich structures could be destroyed.The variation of the current was directly corresponding to the amount of the ATP.The application of SiO2nanoparticles could effectively reduce the background and increase the sensitivity of the biosensor.The calibration curve of ATP was obtained in the range of 0.025-0.900 nmol/L with the detection limit of 13 pmol/L (S/N=3).Also,the biosensor exhibited a good specificity.Besides,the sensor was constructed easily and possessed excellent regeneration ability.The proposed biosensor was applied in detection of real sample such as mice blood.Therefore,the proposed ATP-sensing biosensor could be expected to be applied in clinical,pharmaceutical and environmental detection.

Effect of artificial musk aqueous extract on the expressions of inflammatory mediators released from lipopolysaccharide-stimulated RAW264.7 cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To identify the anti-inflammatory effects of artificial musk aqueous extract(AME)on lipopolysaccharide-stimulated cytokines secreted or released by RAW264.7 cells.</p><p><b>METHODS</b>Cytokines including interleukin(IL)-6,IL-10,and tumor necrosis factor Α were determined using cytokine enzyme-linked immunosorbent assay kits.</p><p><b>RESULT</b>Compared with model group,the levels of major cytokines such as tumor necrosis factor Α,IL-6,and IL-10 significantly decreased in different AME groups in a dose-dependent manner.</p><p><b>CONCLUSION</b>In lipopolysaccharide-stimulated RAW264.7 macrophages,AME can remarkably inhibit the release of inflammatory cytokines and thus exerts its anti-inflammatory effects.</p>

Synthesis of calophyllolide analogue and its preliminary anti-inflammatory activity / 药学学报

A new pyranocoumarin as an analogue of calophyllolide, compound 6, is firstly prepared and reported to have potent anti-inflammatory activity on carrageenin-induced edema in rats.

Effects of dihydroxy-stilbene compound Vam3 on airway inflammation, expression of ICAM-1, activities of NF-kappaB and MMP-9 in asthmatic mice / 药学学报

The aim of the present study is to investigate the effects of Vam3 which is one of the dihydroxystilbene compounds on expressions of ICAM-1 in the lungs of OVA-induced asthmatic mice and the mechanisms of anti-airway inflammation. Balb/c mice were challenged with OVA inhalation. Lung tissues were stained with Mayer's hematoxylin and eosin for histopathologic examination. The expression of ICAM-1 in the lungs of mice was analyzed by Western blotting and immunohistochemistry method. The NF-kappaB activities were detected by NF-kappaB-luc reporter genetic transient transfection method. The activities of MMP-9 induced by LPS, TNF-alpha and PMA in THP-1 cells were determined by gelatin zymography method. The results showed that Vam3 could inhibit the expression of ICAM-1 in the OVA-induced mouse model. In addition, Vam3 could significantly suppress the activities of NF-kappaB in A549 cells and MMP-9 in THP-1 cells induced by LPS, TNF-alpha and PMA. These results suggested that Vam3 could alleviate the asthmatic inflammation by decreasing ICAM-1 expression in asthmatic mice, down regulating NF-kappaB and MMP-9 activities. Compound Vam3 showed inhibitory effects on inflammatory signal pathways involved in asthma.

Synthesis and anti-inflammatory activity of hydroxylated E,E-1-(3'-indolyl)-5-(substituted phenyl)-1,4-pentadien-3-one derivatives / 药学学报

<p><b>AIM</b>A series of new 1,4-pentadien-3-one derivatives were synthesized to search for new Eight novel hydroxylated non-steroidal anti-inflammatory drugs (NSAIDs) with potent activity.</p><p><b>METHODS</b>E,E-1-(3'-indolyl)-5-( substituted phenyl)-1,4-pentadien-3-one derivatives were synthesized by means of aldol condensation and characterized by 1H NMR, ESI-MS and element analysis. Their anti-inflammatory activity in vitro were evaluated.</p><p><b>RESULTS</b>Preliminary in vitro pharmacological tests showed that all compounds exhibited anti-inflammatory activity.</p><p><b>CONCLUSION</b>Compounds 4d and 4e exhibited potent anti-inflammatory activity and their anti-inflammatory activity was comparable to resveratrol, and were worthy of further study.</p>

Ginsenoside-Ro enhances cell proliferation and modulates Th1/Th2 cytokines production in murine splenocytes / 药学学报

<p><b>AIM</b>To study the effects of ginsenoside-Ro on cell proliferation and cytokine production in murine splenocytes.</p><p><b>METHODS</b>The effect of ginsenoside-Ro on murine splenocytes proliferation was studied using [3H] thymidine incorporation assay. Effects of ginsenoside-Ro on the production of cytokines interleukin-2 (IL-2), interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) from murine splenocytes were detected by ELISA method. Effects of ginsenoside-Ro on mRNA level of Th1 cytokine IFN-gamma and Th2 cytokine IL-4 were evaluated by reverse transcription polymerase chain reaction (RT-PCR) analysis.</p><p><b>RESULTS</b>Ginsenoside-Ro showed no mitogenic effect on unstimulated murine splenocytes. It enhanced the proliferation of Con A-induced murine splenocytes and the production of IL-2 at concentrations of 1-10 micromol x L(-1). Moreover, ginsenoside-Ro increased the production and expression of Th2 cytokine IL-4 and decreased the production and expression of Th1 cytokine IFN-gamma in Con A-induced murine splenocytes at concentrations of 2-10 micromol x L(-1).</p><p><b>CONCLUSION</b>Ginsenoside-Ro showed immunomodulatory effects by regulating the production and expression of Th1/Th2 cytokines in murine splenocytes.</p>

Effect of anti-inflammatory drugs on the NF-kappaB activation of HEK293 cells / 药学学报

<p><b>AIM</b>To investigate the regulatory effects of various anti-inflammatory drugs on both endogenous and TNFalpha-induced NF-kappaB activation as well as the relative biological activity.</p><p><b>METHODS</b>HEK293 cells were cultured in 96-well plate and 6-well plate, treated with meloxicam, indomethacin, dexamethasone and hydrocortisone, without or with 10 ng.mL(-1) TNFalpha for 24 hours. Then cell proliferation was measured by MTT and cell apoptosis was analyzed by pI stain-flow cytometry. HEK293/ kappaB-luc cells transfected stably with pElam-kappaB-luc vector, were cultured in 96-well plate and treated as above. Equal amounts of cell lysates were tested for luciferase activity which represents NF-kappaB activation.</p><p><b>RESULTS</b>Endogenous NF-kappaB activation was present in HEK293 cells and its level can be increased about 2 times by 10 ng.mL(-1) TNFalpha-induction. Dexamethasone (1 x 10(-8) mol.L(-1)) and meloxicam (1 x 10(-7) - 1 x 10(-6) mol.L(-1)) can decrease both endogenous and TNFalpha-induced NF-kappaB activation. Hydrocortisone (1 x 10(-9) mol.L(-1)) increases endogenous NF-kappaB activation but decreases TNFalpha-induced one significantly. No influence of indomethacin on endogenous NF-kappaB activation was observed. However, its influence on TNFalpha-induced NF-kappaB activation is needed for further study. Cell apoptosis was observed after treatment with TNFalpha and 1 x 10(-8), 1 x 10(-6) mol.L(-1) dexamethasone and 1 x 10(-7) mol.L(-1) indomethacin, or only with dexamethasone. No significant effect of these anti-inflammatory drugs on cell proliferation was observed.</p><p><b>CONCLUSION</b>Various anti-inflammatory drugs differ in their ability to regulate NF-kappaB activation in HEK293 cells, which indicates that NF-kappaB activation might be a potential useful target to study mechanism and for drug screening.</p>

Effects of musk glucoprotein on chemotaxis of polymorphonuclear leukocytes in vivo and in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effects of Musk glucoprotein on chemotaxis of Polymorphonuclear leukocytes(PMN).</p><p><b>METHOD</b>The chemotaxis of PMN in abdominal cavity in rat induced by carboxymethyl cellulose(CMC) was used as an in vivo animal model and in in vitro it was evaluated by Boyden chamber. The concentration of cytosolic free Ca2+ was quantitated with the fluorescent Ca2+ indicator Fura-2.</p><p><b>RESULT</b>The water extract of Musk at dose of 5, 20, 80 mg.kg-1 (s.c.) significantly inhibited the chemotaxis of PMN in rat; Musk-1 at concentration of 1-100 micrograms.mL-1 can significantly inhibit the chemotaxis of rabbit PMN in vitro; Musk-1 at concentration of 1-100 micrograms.mL-1 can significantly inhibit the increase of cytosolic Ca2+ concentration in PMN of rat.</p><p><b>CONCLUSION</b>Part of mechanisms underlying antiinflammatory action of Musk is to inhibit the chemotaxis of PMN.</p>

Inhibition of dexamethasone, indomethacin and resveratrol on matrix metalloproteinase-9 and the mechanism of inhibition / 药学学报

<p><b>AIM</b>To investigate the expression of matrix metalloproteinase-9 (MMP-9) in mouse ears induced with croton oil and the inhibitory effect of dexamethasone, indomethacin and resveratrol on MMP-9 expression, and further explore the relationship between anti-inflammation and MMP-9 inhibition of these three medicines.</p><p><b>METHODS</b>Immuno-histochemistry was used to detect the expression of MMP-9 in mouse ears. Expression of MMP-9 in U937 cells was analyzed by gelatin zymography.</p><p><b>RESULTS</b>Mouse ear edema induced with croton oil was inhibited significantly by dexamethasone and indomethacin at the dose of 10 mg.kg-1 and resveratrol at 50 mg.kg-1 administered subcutaneously. The inhibitory rate was 76.2% (P < 0.001), 56.7% (P < 0.001) and 36.9% (P < 0.001) respectively. The MMP-9 expression increased in mouse ears induced with croton oil and inhibited by dexamethasone, indomethacin and resveratrol at above doses. Gelatin zymography results showed that MMP-9 expression in U937 cells increased significantly after exposed to PMA at 1 x 10(-8) mol.L-1 (P < 0.001); MMP-9 expression induced with phorbol myristate acetate(PMA) was inhibited by dexamethasone at 1 x 10(-9), 1 x 10(-7) and 1 x 10(-5) mol.L-1, indomethacin at 1 x 10(-6) and 1 x 10(-5) mol.L-1 and resveratrol at 1 x 10(-6) and 1 x 10(-5) mol.L-1.</p><p><b>CONCLUSION</b>The inhibition of MMP-9 expression may be one of the anti-inflammatory mechanisms of dexamethasone, indomethacin and resveratrol.</p>

Effect of indomethacin on expression of interleukin-6 caused by lipopolysaccharide in rheumatoid arthritic patients' synoviocyte / 药学学报

<p><b>AIM</b>To study the effects of indomethacin on interleukin-6 (IL-6) expression stimulated with lipopolysaccharide (LPS) in rheumatoid arthritic patients' synoviocyte.</p><p><b>METHODS</b>Fibroblast-like cells (FLS) from rheumatoid arthritic patients' joint tissue were cultured for 24 h and incubated 24 h with LPS (1 mg.L-1) or the supernatant of U937 cells stimulated by LPS (1 mg.L-1). After indomethacin or dexamethasone added into the supernatant of U937 cells, FLS was incubated with the super natant for 24 h. The expression of IL-6 protein was detected by radioimmunoassay. The mRNA expression of IL-6 was accessed by RT-PCR.</p><p><b>RESULTS</b>LPS did not obviously affect the growth of FLS, and the protein secretion and mRNA expression of IL-6 were not changed in FLS treated with LPS. The IL-6 secretion and IL-6 mRNA expression were significantly increased in FLS cultured with the supernatant from U937 cell treated with LPS. Indomethacin at concentrations of 1 x 10(-7)-1 x 10(-5) mol.L-1 obviously inhibited the protein secretion and mRNA expression of IL-6 in FLS cultured with the supernatant from U937 cell stimulated with LPS, and the inhibitory effects increased as the concentrations of indomethacin increased.</p><p><b>CONCLUSION</b>Indomethacin can inhibit the increase of IL-6 expression caused by supernatant of U937 cells stimulated with LPS in FLS.</p>

Effect of lipopolysaccharide on expression of matrix metalloproteinase-9 in human synoviocyte from patients with rheumatoid arthritis / 药学学报

<p><b>AIM</b>To study the effects of lipopolysaccharide (LPS), the supernatant of U937 cells stimulated with LPS and dexamethasone on matrix metalloproteinase-9 (MMP-9) expression in the synoviocyte from patients with rheumatoid arthritis(RA).</p><p><b>METHODS</b>Fibroblast-like cells (FLS) from the joint tissue of patients with rheumatoid arthritis were cultured and incubated for 24 h with LPS (1 mg.L-1) or the supernatant of U937 cells stimulated with LPS (1 mg.L-1) for 24 h. Dexamethasone was added to the supernatant of U937 cells and FLS was incubated for 24 h. The activity of MMP-9 was analyzed by gelatin zymography. Protein expression of MMP-9 was detected by Western blot using special polyclonal antibodies. The mRNA expression of MMP-9 was detected by RT-PCR.</p><p><b>RESULTS</b>The expression of MMP-9 was not markedly changed in FLS treated with LPS. The MMP-9 activity, MMP-9 secretion and MMP-9 mRNA expression were significantly increased in FLS cultured with the supernatant from U937 cell treated with LPS. Dexamethasone markedly inhibited the activity, protein secretion and mRNA expression of MMP-9 in FLS cultured with the supernatant from U937 cell stimulated with LPS, and the inhibitory effects were increased as the concentration of dexamethasone increased.</p><p><b>CONCLUSION</b>LPS did not directly affect the expression of MMP-9 in FLS, but it was found to indirectly cause the increase of MMP-9 expression in FLS by stimulating U937 cell. Dexamethasone was found to inhibit this increase of MMP-9 expression.</p>

Effect of lipopolysaccharide on expression of interleukin-6 in human synoviocyte from patients with rheumatoid arthritis / 药学学报

<p><b>AIM</b>To study the effects of lipopolysaccharide (LPS), the supernatant of U937 cells stimulated with LPS and dexamethasone on interleukin-6 (IL-6) expression in the synoviocyte from patients with rheumatoid arthritis (RA).</p><p><b>METHODS</b>Fibroblast-like synoviocytes (FLS) from the joint tissue of patients with rheumatoid arthritis were cultured and incubated for 24 h with LPS (1 mg.L-1) or the supernatant of U937 cells stimulated with LPS (1 mg.L-1) for 24 h. Dexamethasone was added to the supernatant of U937 cells and FLS was incubated for 24 h. The expression of IL-6 protein was detected by radioimmunoassay. The mRNA expression of IL-6 was accessed by RT-PCR.</p><p><b>RESULTS</b>The growth of FLS was not markedly affected by LPS, and the protein secretion and mRNA expression of IL-6 were not markedly changed in FLS treated with LPS. The IL-6 secretion and IL-6 mRNA expression were significantly increased in FLS cultured with the supernatant from U937 cell treated with LPS. Dexamethasone markedly inhibited the protein secretion and mRNA expression of IL-6 in FLS cultured with the supernatant from U937 cell stimulated with LPS. The inhibitory effects were increased as the concentration of dexamethasone increased.</p><p><b>CONCLUSION</b>LPS was not shown to directly affect the expression of IL-6 in FLS, but it indirectly causes the increase of the IL-6 expression in FLS by stimulating U937 cell. Dexamethasone can inhibit this increase of the IL-6 expression.</p>

Inhibitory effect of meloxicam on human polymorphonuclear leukocyte adhesion to human synovial cell / 药学学报

<p><b>AIM</b>To investigate the effect of meloxicam on human polymorphonuclear leukocyte (PMN) adhesion to human synovial cell (HSC), and to explore its mechanism.</p><p><b>METHODS</b>MTT colorimetry was used to determine the adhesion effect of PMN to HSC. Cell-ELISA and RT-PCR methods were used to determine the expression of ICAM-1 and VCAM-1. Nuclear transcription factor-kappa B (NF-kappa B) was measured by electrophoretic mobility shift assay (EMSA) method.</p><p><b>RESULTS</b>Meloxicam was found to effectively inhibit TNF-alpha (50 u.mL-1 for 12 h) and IL-1 beta (50 u.mL-1 for 12 h)-induced adhesion of PMN to HSC (IC50 3.38 x 10(-7) mol.L-1 and 3.56 x 10(-6) mol.L-1, respectively) in a concentration-dependent manner. ICAM-1 protein and mRNA expression induced by TNF-alpha (50 u.mL-1) were inhibited by meloxicam at 1 x 10(-6)-1 x 10(-5) mol.L-1. The activation of NF-kappa B was also inhibited by meloxicam at 1 x 10(-6)-1 x 10(-5) mol.L-1.</p><p><b>CONCLUSION</b>These results suggest that meloxicam inhibit TNF-alpha stimulated PMN-HSC adhesion and expression of ICAM-1 by suppressing the activity of NF-kappa B.</p>