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Objective To explore the effect of blood-brain barrier disruption on expression of AQP-4,through comparing the cell morphology and the expression of aquaporin-4(AQP-4)of cultured astrocytes in medium with and without fetal bovine serum(FBS). Methods Cerebral cortical astrocytes from female Wistar rats were cultured in serum free medium,DMEM supplement-ed with 10% FBS,and serum free medium supplemented with 10% FBS.Phase contrast microscope was used to detect the cell morphology and cell size. Immunofluorescence staining and reverse real-time quantitative poly-merase chain reaction(RT-qPCR)were used to examine the expression of glial fibrillary acidic protein(GFAP), AQP-4 and metabotropic glutamate receptor 5(mGluR5). Results Astrocytes in serum free medium showed extensive process bearing morphology,small body and nucleus,and high refractivity.In contrast,in two kinds of 10% FBS-containing medium,astrocytes were flat with large body and nucleus,weak refractivity,as well as short process.Analysis of immunofluorescence staining and RT-qPCR revealed a down-regulation of GFAP and AQP-4 protein and mRNA expression in two kinds of 10% FBS-con-taining medium, compared with that in serum free medium (P<0.001), however, there was no difference in mGluR5 protein and mRNA expression(P>0.05). Conclusion FBS changed astrocyte morphology and down-regulated the expression of GFAP and AQP-4.
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Objective To study the feasibility of novelty detection model in assessing the subject novelty of medical literature and comparatively analyze the advantages and disadvantages of words-overlap algorithm and co-words-based inverse file frequency quantitative algorithm. Methods Two novelty detection models were established for the 8 research subjects in PubMed-covered literature. The feasibility of two novelty detection models in assessing the subject novelty of medical literature was assessed according to the subject novelty of literature analyzed by experts, ROC curves and AUC values. Results Words-overlap algorithm showed that the fluctuating amplitude of subject novelty was rather high, which can thus reflect the difference between the contents in literature on the data. ROC curves and AUC values-based analysis revealed a high accuracy of words-overlap algorithm for judging the novelty of literature while co-words-based inverse file frequency quantitative algorithm displayed a low accuracy for judging the novelty of literature. Conclusion The novelty of literature detected with the two novelty detection methods is moderately related. The mean novelty value detected with the two novelty detection methods is of statistical signifi-cance. However, the novelty of literature detected with words-overlap algorithm is higher than that detected with co-words-based inverse file frequency quantitative algorithm.
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Gentiana L. is the largest genera in Gentianaceae. Plants in Gentiana L. contain a lot of chemical constituents with pharmacological activities, especially iridoids. Iridoids has good hepatoprotective, anti-inflammatory, antipyretic, antiviral, and other pharmacological effects. This article summarizes the research progress of iridoids in plants from Gentiana L., which provide references for the studies of chemical constituents in Gentiana L. and the exploitation of medicinal resources.
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<p><b>OBJECTIVE</b>To investigate the role of extracellular signal-regulated kinase1/2 (ERK1/2) pathway in the regulation of aquaporin 4 (AQP4) expression in cultured astrocytes after scratch-injury.</p><p><b>METHODS</b>The scratch-injury model was produced in cultured astrocytes of rat by a 10-μL plastic pipette tip. The morphological changes of astrocytes and lactate dehydrogenase (LDH) leakages were observed to assess the degree of scratch-injury. AQP4 expression was detected by immunofluorescence staining and Western blot, and phosphorylated-ERK1/2 (p-ERK1/2) expression was determined by Western blot. To explore the effect of ERK1/2 pathway on AQP4 expression in scratch-injured astrocytes, 10 µmol/L U0126 (ERK1/2 inhibitor) was incubated in the medium at 30 min before the scratch-injury in some groups.</p><p><b>RESULTS</b>Increases in LDH leakage were observed at 1, 12, and 24 h after scratch-injury, and AQP4 expression was reduced simultaneously. Decrease in AQP4 expression was associated with a significant increase in ERK1/2 activation. Furthermore, pretreatment with U0126 blocked both ERK1/2 activation and decrease in AQP4 expression induced by scratch-injury.</p><p><b>CONCLUSION</b>These results indicate that ERK1/2 pathway down-regulates AQP4 expression in scratch-injured astrocytes, and ERK1/2 pathway might be a novel therapeutic target in reversing the effects of astrocytes that contribute to traumatic brain edema.</p>
Subject(s)
Animals , Rats , Aquaporin 4 , Metabolism , Astrocytes , Metabolism , Butadienes , Cells, Cultured , Down-Regulation , Enzyme Activation , Extracellular Signal-Regulated MAP Kinases , Metabolism , MAP Kinase Signaling System , Nitriles , Rats, Wistar , Skin , Wounds and InjuriesABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of Bushen Antai Recipe (BAR) on expression of prostaglandin I2 (PGI2) and its nuclear receptor peroxisome proliferators-activated receptor delta (PPARdelta) at implantation site in mice with blastocyst implantation dysfunction.</p><p><b>METHODS</b>Pregnant mice were divided into three groups randomly, the normal group, the model group and the BAR group. The pregnant uterus of all mice was cut off on the 4th (D4), 5th (D5), 6th (D6) and 8th (D8) day of pregnancy for determining the PGI2 expression with radio immunoassay; and the mRNA and protein expression of PPARdelta with RT-PCR and immunohistochemistry at implantation site.</p><p><b>RESULTS</b>PGI2 expression in the model group was obviously lower than that in the normal group (P < 0.01), and also lower than that in the BAR group (P < 0.01), while the index was insignificantly different between the normal and the BAR group. Compared with the normal group, the expression of PPARdelta in the model group was delayed temporally and spatially (P < 0.05), while that in the BAR group was not significantly different.</p><p><b>CONCLUSION</b>BAR can improve the implantation in mice with blastocyst implantation dysfunction through promoting the PGI2 expression and its nuclear receptor PPARdelta at implantation site.</p>
Subject(s)
Animals , Female , Male , Mice , Pregnancy , Drugs, Chinese Herbal , Pharmacology , Embryo Implantation, Delayed , Endometrium , Metabolism , Epoprostenol , Immunohistochemistry , PPAR delta , Genetics , RNA, Messenger , Genetics , Radioimmunoassay , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Recent studies showed that Eph/Ephrin tyrosine kinase family plays an important role in the development and functional maintenance of the nervous system, but its function in the sympathetic nervous system is still obscure. In the present study, we examined the effect of Eph/Ephrin-B1 signaling on the whole-cell currents mediated by either alpha7 or alpha3-nicotinic acetylcholine receptors (nAChRs) in acutly dissociated ciliary ganglion (CG) neurons. Firstly, we detected the effect of Ephrin-B1 on nAChRs currents. The neurons were randomly divided into control group, Ephrin-B1Fc-treated group that was stimulated by recombinant Ephrin-B1Fc, IgG-treated group, and Ephrin-B1-treated group. Secondly, we studied the regulatory mechanism of Ephrin-B1Fc on nAChRs currents. The neurons were randomly divided into control group, Ephrin-B1Fc-treated group, PP2 (inhibitor of Src tyrosine kinase) or PD98095 (antagonist of mitogen-activated protein kinase)-treated group, Ephrin-B1Fc + PP2 or PD98095-treated group. The results showed that there was no significant difference between the currents in control group, IgG-treated group and Ephrin-B1-treated group, but Ephrin-B1Fc significantly suppressed both alpha3-nAChRs and alpha7-nAChRs-mediated currents (P=0.002, P=0.003). Pretreatment with PP2 or PD98095 could partially rescue the Ephrin-B1Fc-induced suppression of currents mediated by alpha3-nAChRs or alpha7-nAChRs respectively. These results suggest that the Eph/Ephrin-B1 signaling may inhibit alpha3-nAChRs and alpha7-nAChRs-mediated currents on CG neurons, involving Src tyrosine kinase and mitogen-activated protein kinase signaling in the regulation of sympathetic nervous system.
Subject(s)
Ephrin-B1 , Metabolism , Ganglia, Parasympathetic , Mitogen-Activated Protein Kinases , Metabolism , Neurons , Receptors, Nicotinic , Metabolism , Signal Transduction , alpha7 Nicotinic Acetylcholine Receptor , Metabolism , src-Family Kinases , MetabolismABSTRACT
<p><b>AIM</b>To investigate the protective effect of mGluR1 antisense oligonucleotides and mGluR5 antisense oligonucleotides on impairment of cultured mouse cerebral cortical neurons induced by sodium glutamate (Glu).</p><p><b>METHODS</b>Neuron damage induced by Glu as well as the action of mGluR1 antisense oligonucleotides and mGluR5 antisense oligonucleotides were measured by determining the leakage of lactate dehydrogenase (LDH) from neurons. Immunocytochemistry method was used to detect the expression of anti-mGluRl a and anti-mGluR5. Morphological changes of primary cortical neurons were observed by phase contrast microscope.</p><p><b>RESULTS</b>Following the exposure of the cells to 0.1 mmol x L(-1) Glu for 15 min, LDH leakage from neurons increased. mGluR1 antisense oligonucleotides and mGluR5 antisense oligonucleotides(6 or 8 micromol x L(-1)) as well as 50 micromol x L(-1) LY367385 reduced the LDH leakage. mGluR1alpha and mGluR5 immunopositive cells showed in cultured neurons.</p><p><b>CONCLUSION</b>The protective effects of mGluR1 antisense oligonucleotides and mGluR5 antisense oligonucleotides on neurons damaged by Glu may relate to antagonizing mGluR1a or mGItlR5.</p>