The peripheral benzodiazepine receptor (TSPO/PBR) is highly conserved among different species but with perplexing biochemical functions. Multiple ligands of TSPO show commendable regulatory ...activities in lots of biological functions, such as neuro‐protection, cholesterol transport, and so on. These researches support that TSPO may be a potential target for disease treatment and drug development. Previous studies have shown that its ligands benzodiazepines show a satisfactory effect on melanogenic promotion. However, the potential application of TSPO in drug development for pigmentary disorder needs further investigation. In this study, we confirmed the melanogenesis induction of TSPO ligand, Ro5‐4864 in mouse melanoma cell lines, human skin tissue, and zebrafish embryos by inducing melanin synthesis and melanosome transport. Molecular genetics and pharmacological studies showed that TSPO deficiency did not affect melanin production in B16F10 cells and zebrafish embryos, nor did it affect the melanin promotion effect of Ro5‐4864. Whether or not TSPO exists, the expression of lots of melanogenesis‐related proteins, such as TYR, TRP‐1, DCT, Mlph, and Rab27 was upregulated with the Ro5‐4864 administration. These results indicated that Ro5‐4864 induces melanogenesis in a TSPO‐independent manner, which is inconsistent with previous research. This research is a reminder that we need to be very careful during target validation in drug development.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
During further chemical and biological investigations of Picrasma quassioides BENNET, four new bis-β-carboline alkaloids, quassidines E—H (1—4), and three new β-carboline alkaloids, ...canthin-16-one-14-butyric acid (5), 3-(1,1-dimethoxylmethyl)-β-carboline (6), and 6,12-dimethoxy-3-formyl-β-carboline (7), were isolated from its anti-inflammatory CHCl3-soluble fraction. Structures of new compounds were elucidated and characterized by MS and NMR analysis. A plausible biogenetic pathway for quassidine E (1), the first bis-β-carboline alkaloid in which a canthin-6-one moiety and a β-carboline moiety were connected together by a single carbon–carbon bond from the nature, was proposed. Quassidines E—G (1—3) showed potent inhibitory activity on the production of nitric oxide (NO), tumor necrosis factor α (TNF-α), or interleukin 6 (IL-6) in mouse monocyte-macrophage RAW264.7 cells stimulated by lipopolysaccharide (LPS). Analysis of anti-inflammatory activity of all β-carboline and bis-β-carboline alkaloids from P. quassioides showed that the carbonyl groups or double carbon–carbon bonds at C-14 for β-carbolines and C-14′ for bis-β-carbolines were bioactive groups for their in vitro anti-inflammatory activity. Structure–activity relationship of these compounds on inhibitory activity of the three inflammatory cytokines was discussed.
Abstract Repellent guidance molecules provide targeting information to outgrowing axons along predetermined pathways during development. These molecules may also play a role in synaptic ...reorganization in the adult brain and thereby promote epileptogenesis. Our aim was to investigate the expression of Slit2, one of repellent guidance molecules, in temporal lobe epileptic foci from epileptic patients and experimental animals. Thirty-five temporal neocortex tissue samples from patients with intractable temporal lobe epilepsy (TLE) and fifteen histological normal temporal lobes from controls were selected. Fifty-four Sprague–Dawley rats were divided randomly into six groups, including five groups with epilepsy induced by lithium–pilocarpine administration and one control group. Temporal lobe tissue samples were taken from rats at 1, 7, 14, 30, and 60 days post-seizure and from controls. Expression of Slit2 was assessed by immunohistochemistry, immunofluorescence, and Western blot analysis. Slit2 was mainly expressed in neurons in human controls and in both neurons and astrocytes in TLE patients. Slit2 expression was significantly higher in TLE patients as compared with the controls. Slit2-positive cells were mainly neurons in the rat temporal lobe tissues of the control group, the acute period group, and the latent period group, while the Slit2-positive cells were mainly astrocytes in chronic phase. Compared with controls, Slit2 expression in animals in the TLE group gradually decreased from days 1 to 14 post-seizure, but then increased over the levels seen in controls, to peak levels at days 30 and 60. These results suggest that Slit2 may play an important role in the pathogenesis of TLE.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
AIM： To investigate the mechanism of interleukin （IL）-6 secretion through blocking the IL-17A/IL-17A recepto （IL-17RA） signaling pathway with a short hairpin RNA （shRNA） in hepatic stellate cells ...（HSCs） in vitro . METHODS： HSCs were derived from the livers of adul male Sprague-Dawley rats. IL-6 expression was evalu ated using real-time quantitative polymerase chain reaction and enzyme linked immunosorbent assay. The phosphorylation activity of p38 mitogen activated pro tein kinases （MAPK） and extracellular regulated pro tein kinases （ERK） 1/2 upon induction by IL-17A and suppression by IL-17RA shRNA were examined using Western blotting.RESULTS： IL-6 expression induced by IL-17A was significantly increased compared to control in HSCs （P 0.01 in a dose-dependent manner）. Suppression of IL17RA using lentiviral-mediated shRNA inhibited IL-6 expression induced by IL-17A compared to group with only IL-17A treatment （1.44 ± 0.17 vs 4.07 ± 0.43, P 0.01）. IL-17A induced rapid phosphorylation of p38 MAPK and ERK1/2 after 5 min exposure, and showed the strongest levels of phosphorylation of p38 MAPK and ERK1/2 at 15 min in IL-17A-treated HSCs. IL-6 mRNA expression induced by IL-17A （100 ng/mL） for 3 h exposure was inhibited by preincubation with specific inhibitors of p38 MAPK （SB-203580） and ERK1/2 （PD-98059） compared to groups without inhibitors preincubation （1.67 ± 0.24, 2.01 ± 0.10 vs 4.08 ± 0.59, P 0.01）. Moreover, lentiviral-mediated IL-17RA shRNA 1 inhibited IL-17A-induced IL-6 mRNA expression compared to random shRNA in HSCs （1.44 ± 0.17 vs 3.98 ± 0.68, P 0.01）. Lentiviral-mediated IL17RA shRNA 1 inhibited phosphorylation of p38 MAPK and ERK1/2 induced by 15 min IL-17A （100 ng/mL） exposure. CONCLUSION： Down-regulation of the IL-17RA receptor by shRNA decreased IL-6 expression induced by IL-17A via p38 MAPK and ERK1/2 phosphorylation in HSCs. Suppression of IL-17RA expression may be a strategy to reduce the inflammatory response induced by IL-17A in the liver.
Microsomal casein kinase II (mCKII) is a membrane-bound enzyme present in the microsomal fractions of ROS 17/2.8 osteoblast-like cells. It phosphorylates acidic matrix phosphoproteins such as ...phosphophoryn and osteopontin. Addition of 1.0% Nonidet P-40 facilitates extraction of the optimum amount of detergent -solubilized and -activated enzyme from microsomal fractions. mCKII was partially purified over 3000-fold by sequential chromatography over DEAE-cellulose and heparin-agarose. SDS-polyacrylamide gels, showed that mCKII contained 43 kDa and 31 kDa polypeptides, corresponding to the α- and β- subunits of the enzyme, respectively. The α subunit was identified by anti-CKII antiserum and the β subunit, by its ability to undergo autophosphorylation. The enzyme was inhibited by 50% with 0.4 μg/ml heparin and stimulated by 100% with 1.0 mM spermine when casein was used as a substrate. The phosphorylation of phosphophoryn was reduced to 50% by 0.8 μg/ml heparin, but was increased to 2-2.5 fold by 5 to 15 mM spermine, which may be due to substrate-directed effects. Kinetic analysis showed that the apparent Km values for phosphophoryn (0.39 μM) and for osteopontin (2.1 μM) were lower than that for casein (21.3 μM). Vmax values of phosphophoryn and osteopontin were 2.2-fold and 4.6-fold higher than that of casein. Using the ratio Vmax/Km as a measure of kinetic specificity, osteopontin and phosphophoryn appear to be the more specific substrates than casein for mCKII. Thus, both proteins can be considered as physiological substrates for mCKII.
The microRNA-21 （miR-21） is known to play a major role in cancer progression; however, its function in the cardiovascular system appears to be even more complex and conflicting. To characterize ...miR-21 expression in the plasma of individuals with or without metabolic syndrome （MetS）, 58 MetS cases and 96 non-MetS controls were investigated.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
In order to improve the precision of gas emission forecasting,this paper proposes a new forecasting model based on Particle Swarm Optimization (PSO).PSO is a novel random optimization method which ...has extensive capability of global optimization.In the model, PSO is used to optimize the weight,width and center of RBF neural network and the optimal model is applied to forecast gas emission.The diversified factors analysised with grey correlation,MATLAB is employed to implement the model for gas emission forecasting.The simulation results show that the gas emission model optimized by PSO is more accurate than the traditional RBF model．
Objective Waist circumference, as a brief indicator of visceral obesity, is associated with multi-metabolic disorders and cardiovascular diseases. The present study was aimed to find out the ...relationship between waist circumference and carotid intima media thickness （C-IMT）, as well as the best waist circumference cutoff for identifying C-IMT elevation in Chinese male patients with newly-diagnosed diabetes. Methods Five hundred and seventy-eight patients from Department of Endocrinology and Metabolism in Shanghai Sixth People＇s Hospital affiliated to Shanghai Jiao Tong University were enrolled. Both physical examination （for measurement of waist circumference） and carotid ultrasonography （for measurement of C-IMT） were performed. Results After grouping according to the quartiles of C-IMT, the waist circumference increased across all its quartiles. The waist circumference in 3rd and 4th quartiles （90.7_＋9.8 cm and 90.8＋9.6 cm） was significant higher than in 1st and 2nd quartiles （P〈0.05）. When subjects were divided into 4 groups according to waist circumference, the C-IMT of subjects with waist circumference 90-95 cm was significant higher than that of subjects with waist circumference 85-90 cm and less than 85 cm respectively （P〈0.05）. Both spearman and partial correlation analysis showed that C-IMT was positively correlated with waist circumference （P〈0.01）. C-IMT was found significantly elevated with the increase of waist circumference. Multiple stepwise regression analysis showed that waist circumference was one of the independent risk factors of C-IMT. After an average of 2.23_＋0.85 years follow up, there was a significant elevation of C-IMT in the group with baseline waist circumference over 90 cm P〈0.05）, while no significant difference was detected in the group with baseline waist circumference less than 90 cm （P=0.27）. Logistic regression showed that baseline waist circumference over 90 cm was associated with a relative risk to C-IMT elevation of 1.132 （95% CI 1.043-1.431, P〈0.05）. Conclusion Among newly-diagnosed diabetic male patients, waist circumference over 90 cm not only reflects sub-clinical atherosclerosis in early stage, but also predicts the progression of atherosclerosis.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The potential of epithelial cells to degrade interstitial collagen was studied by culturing human masticatory mucosa on decalcified dentin matrix. Morphological changes were observed in the ...underlying collagen substratum and in the connective tissue of the explant. Degradation of the substratum was initiated two days after the first contact with epithelial cells exhibiting basal cell markers. Electron microscopic studies confirmed extensive collagen degradation in the vicinity of these cells. No collagen degradation was observed underneath the connective tissue portion of the explant. Experiments in which the explant was partially separated from the underlying substratum by a filter further showed that connective tissue was apparently not involved in the collagen degradation by the epithelial cells. Lysis of connective tissue of the explant was observed in association with epithelial cells that showed a disrupted basal lamina and release of vesicular material from the exposed cell membrane. Collagen fibers were visible inside some epithelial cells suggesting intracellular collagenolysis. Primary cultures of human gingival epithelial cells and porcine periodontal ligament epithelial cells (epithelial cell rests of Malassez) that expressed similar basal cell cytokeratins as the active cells of the mucosal explants secreted collagenase, gelatinase and TIMP to the culture medium. They also contained acid collagenolytic proteinases. When cultured on a porous polycarbonate membrane the epithelial cells secreted collagenolytic enzymes from the pores at cell membrane sites lacking basal lamina. These results provide evidence that proliferating basal epithelial cells have a strong capacity for collagen degradation. It seems that the absence of basement membrane is the signal for these cells to secrete matrix degrading enzymes.
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