ABSTRACT
Objective To investigate the effect and the potential mechanisms of low molecular polysaccharide from agaricus blazei (LMPAB) on H2O2-induced oxidative injury in hippocampal neuronal cells of rats.Methods Hippocampal neuronal cells were isolated from SD rats (24 h) and grew in culture.Cultured cells were divided into normal control group (added the same amount of nutrient solution), model control group (added 500 μmol·L-1H2O2 solution) and LMPAB high, medium, low dose groups (added 20,10,5 mg·L-1 LMPAB solution, respectively, then added 500 μmol·L-1 H2O2 solution each).The hippocampal neuron cell activity was detected with MTT method.The hippocampus neuron mitochondrial membrane potential (MMP) was detected by flow cytometry.According to the reagent instruction methods, malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities were detected.Results The activities of cell, CAT, SOD, GSH-PX and MMP in normal control group and the LMPAB high dose group were significantly higher than those of model control group (P<0.01);The content of MDA in normal control group and LMPAB high dose group was significantly lower than that of model control group (P<0.01).Conclusion The protective effect of LMPAB on hippocampal neurons with H2O2-induced injury may be related with the mechanism of enhancing the neuronal antioxidative capacity.
ABSTRACT
Aim To investigate the effects of midazolam on porcine isolated coronary artery rings pre-contracted by potassium chloride(KCl)and the possible mechanism.Methods The vessel tension recorder system was used.Isotonic tension of porcine isolated coronary artery rings precontracted by KCl(30 mmol·L-1)was recorded.The vasorelaxing action of midazolam and effects of various drugs were observed in the rings.Results Midazolam(3×10-6～1×10-4 mol·L-1)respectively concentration-dependently reduced the contraction induced by KCl,and there was significant difference between the rings with intact and denude endothelium(P<0.05).On KCl-induced precontraction,midazolam′s relaxation was depressed by L-NAME and the blend of L-NAME and L-Arg(P<0.05),but was not affected by Indo,L-Arg and 1400W.The contraction was not prevented by pretreatment with the inhibitor of Na+/Ca2+ exchanger(KB-R7943).The inhibitor of KATP(Gli)restrained the diastolic function of midazolam(P<0.05),while the inhibitor of BKCa(TEA),Kir(BaCl2),KV(4-AP)had no obvious effect.Conclusions Midazolam produces remarkable vasodilatation on KCl pre-contracted porcine isolated coronary artery rings.Its relaxtion effect is via concentration-dependent and endothelium-dependent mechanisms and relevant to the production of NO.Na+/Ca2+ exchanger is not involved midazolam′s vasodilatation on KCl pre-contracted porcine coronary artery rings.The relaxant mechanism of midazolam may be concerned with KATP.The Kir,BKCa and KV may be not involved.
ABSTRACT
Phage display random heptapeptide library was screened with recombinant P37 in this study. The positive phage clones were identified by ELISA and were sequenced, and the amino acid sequences of the polypeptides displayed on phage were deduced. After GST-polypeptides fusion protein was constructed and expressed, its binding to P37 was determined by GST-pull down and Western blot. After 4 rounds of bio-panning, the enriched positive phage clones were identified by ELISA. Eighteen positive phage clones were sequenced and the peptide sequences were as follows. ACAPKPPWLC (12/18), RPLSIDPWSPHL (3/18), RPLSNDPWSPHL (1/18), QNMMSPIEGVRI (1/ 18) and WAPEKDYMQLMK (1/18). The results from GST-pull down and Western blot showed that peptide RPLSIDPWSPHL could interact with P37. The study will be helpful for identifying the protein reacting with P37.
Subject(s)
Animals , Bacterial Proteins , Metabolism , Base Sequence , Molecular Sequence Data , Mycoplasma hyorhinis , Metabolism , Oligopeptides , Metabolism , Peptide Library , Protein Binding , SwineABSTRACT
<p><b>OBJECTIVE</b>To study the total glucosides of Radix Paeoniae Rubra induced K562 tumor cell apoptosis of signaling pathways and related gene changes.</p><p><b>METHOD</b>By MTT and flow cytometric, real-time quantitatie polymerase chain reaction (PCR) and Western blot methods researched the level of genes and proteins.</p><p><b>RESULT</b>The total glucosides of Radix Paeoniae Rubra could inhibit K562 cell growth by MTT method, there was the relationship of concentration-time between them, TGC prompted K562 cell translocation of phosphatidylserine, may be apoptosis through non-receptor-dependent pathways because caspase-3 mRNA, caspase-9 mRNA and cytochrome C increased, caspase-8 mRNA had no significant change. The expression of Bcl-2 protein and Bcl-X1 protein could decreased, the expression of Bax protein could increased by regulating gene expression.</p><p><b>CONCLUSION</b>TGC induced K562 cell apoptosis that might be through the mitochondria pathway, when cell apoptosis occured, Bcl-2 or Bcl-XI proteins combination of Bax protein would be displacement, then the mitochondrial membrane became permeable to release a series of material, then lead to cell death.</p>
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Apoptosis , Genetics , Caspases , Genetics , Dose-Response Relationship, Drug , Gene Expression Regulation, Neoplastic , Glucosides , Pharmacology , K562 Cells , Paeonia , Chemistry , Signal Transduction , Time FactorsABSTRACT
Objective To connect the content of clinical immunology and inspection with network,so that it is more convenient for medical students to understand and study this course,and the learners’interest in learning is aroused.Methods FrontPage 2003 web page authoring software,as well as animation software(flash) and picture editing software(Photoshop) were used to make a vivid image of the teaching content be shown through the web page,and effectiveness evaluation was done through peer review and students survey about the software.Results Using FrontPage to create web pages and change knowledge into learning pattern.got good results in studetns’self-study,and the"online self-test"section was added,which would more effectively check study effect.There were good student feedbacks.Conclusion Applying this learning pattern to study will play a very good role in deepening students’understanding of curriculum,improving their study efficiency and stimulating their interest in learning.
ABSTRACT
Objective To study the effects of ganoderma applanatum polysaccharides(GAPS) on cell morphology, proliferation and PDGFR -?expression in cell lines MGC - 803 , and to explore its potential mechanisms of anti - tumor of GAPS. Methods Cell morphology was observed by inverting microscope. MTT assay was used to investigate the inhibitory effect of GAPS on MGC -803.Expressions of PDGFR -?was analyzed by flow cytometry (FCW). The fluorescence intensity of expressions of PDGFR -?was observed by fluorescence microscope. Results Cells of GAPS group were irregular shaped and grew poorly. GAPS inhibited the proliferation of MGC -803 cells in dose - dependent and time - dependent manners . After MGC - 803 cells were treated with GAPS for 72h, GAPS could down - regulate expression of PDGFR -?observed by fluorescence microscope which was consistented with the results of FCW with statistic significance difference as compared with control group (P
ABSTRACT
Objective To investigate the effects of quercetin on the proliferation of mammary cancer cell in vitro. Methods Effects of quercetin on the cell proliferation was tested in ER-positive MCF-7 cell and ER-negative MDA-MB231 cell by MTT measurement. And to evaluate the estrogen-like effect of quercetin and its relation with the estrogen-receptor by pure estrogen receptor antagonist ICI 182, 780 as a tool. Cell cycle was examined by flow cytometry. Result Quercetin (10～50 ?mol/L) stimulated proliferation of ER-positive MCF-7 cell compared with solvent control, whereas ER-negative MDA-MB231 cell proliferation effect was inhibited, and the cell cycle was impulsed from G1 to S, DNA synthesizing was inhanced, PI was also increased. The above function on boosting MCF-7 cell proliferation can be inhibited by adding estrogen receptor antagonism ICI182, 780. Conclusion Quercetin has the estrogen-like activities through the estrogen response pathway.
ABSTRACT
Objective Experiment the tumor animal model of the H22 Cancer of the liver mile,study the inhibition effect of chitosan oligosaccharide and bifidobacteria on tumour.Methods the growth of tumour were observed after chitosan oligosaccharide and bifidobacteria were injected into tumor-bearing mice;then the immunify funcfion weremcasured.Results Growth of tumour were inhibited by chitosan oligosaccharide and bifidobacteria,they can increase the content of IL-2 and IFN-?and the weight of spleen and thymus of the mice.Conclusions chitosan oligosaccharide and bifidobacteria could inhibit the growth of tumour,inproue immunity function.
ABSTRACT
Objective:To study TGC on tumor apopotosis and to apply the experimenting basis of TGC's developing and utilizing.Methods:The flow cytometer(FCM) was used for determination apoptosis.The protein expression of Bcl-2 and the mRNA expression of c-myc were evaluated by immunohistochemistry and in situ hybridization.Apoptosis small body were observed by EU.Results:The TGC group appear the Ap apex,the number of cell of G0-G1 period reduce obviously,the cells of S period increase obviously.Bcl-2 and the c-myc mRNA expression levels were decreased in TGC group.The structure of inner membrane was intacted with EU.Chromatin gathered side of cell,the nucleus turns into the nucleus band and nucleus project,there were many cells of the apoptosis and meniscus or the garland shape(the apoptosis corpuscle formatiol) in TGC group.Conclusion:The total glucosides of radix paeoniae induces the tumor cell apoptosising,its mechanism maybe closely related with adjusting expression levels of the Bcl-2 and the c-myc mRNA.