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Aim To investigate the effect of glycyrrhizin on the proliferation, apoptosis and chemosensitivity of lung adenocarcinoma cells by regulating microRNA - 216b-5p (miR-216b-5p). Methods Human lung adenocarcinoma H1299 cells were cultured and randomly divided into control group and glycyrrhizin 10, 20, 40, 60, 80 mg • L
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Objective In this study, electroacupuncture (EA) was used to analyze the expression changes of related proteins in neuroglobin (NGB), PI3K/AKT and apoptotic pathways in the temporal cortex of bilirubin encephalopathy (BE) rats, so as to investigate the therapeutic effect of EA on BE and the relevant mechanism of NGB in this process. Totally 39 seven-day-old SD rats were divided into Sham, BE model and BE+EA groups. The neonatal BE model was established by injecting bilirubin solution (10 μg UCB/g Weight) into the cerebellomedullary cistern, Sham group was injected with the same amount of normal saline. BE rats were treated with EA at Baihui (GV20) and Quchi (LI11) acupoints with the frequency of 2/15 Hz for 15 min. Treatment was performed 12 h before modeling, followed by treatment every 12 h, in a total of three times. HE, Nissl staining and electron microscopy (TEM) were used to observe the pathological and ultrastructural changes of nerve cells in each group. Results showed that EA treatment reduced the damage of cortical neurons of BE rats and increase the number of Nissl bodies. TEM confirmed that EA treatment could alleviate the degree of mitochondria edema. Immunofluorescence staining was used to detect the expression sites and cell types of NGB. Results showed that NGB was mainly expressed in cortical neurons. Western blotting showed that EA treatment increased the expression of NGB, PI3K (p110 alpha), pAKT (Ser473) (P< 0. 05, P< 0. 05 and P< 0. 01, respectively) and the ratio of apoptosis-related protein Bcl-2/Bax (P < 0. 001), decreased the expression of Cleaved Caspase-3 (P< 0. 05) in the temporal cortex of rats. TUNEL staining showed that EA reduced the number of apoptotic cells (BE group 186. 00±13. 86 vs BE+EA group 78. 67±11. 85, P< 0. 01) . This study confirms that EA can promote the expression of NGB in the temporal cortex of BE rats, then activate the PI3K/AKT pathway to exert its neuroprotective function and inhibit the occurrence of apoptosis. EA may become a potential treatment method for BE.
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OBJECTIVE To observe the effects of glycogen synthase 3β (GSK-3β) in the regula-tion of NLRP3 inflammasome activation after acute myocardial infarction (MI) in Sprague Dawley(SD) rats. METHODS Ligation of the left anterior descending (LAD) in SD rats was used to establish an acute myocardial infarction model. SD rats were randomly divided into 3 groups (n=10, each group):sham group,MI group,and MI+SB group:the GSK-3β inhibitor(SB216763)was given 1 h by intrave-nous injection(0.6 mg·kg-1·d-1)before surgery.The serum and heart tissue were collected to measure lactate dehydrogenase (LDH) and IL-1β content and mRNA and protein levels of NLRP3, ASC, Cas-pase-1,IL-1β and GSK-3β after 2 days and 7 days operation,respectively.RESULTS The serum levels of LDH and IL-1β in the MI group were significantly higher than those in the sham group(P<0.01),and the MI+SB group was obviously lower than the MI group(P<0.01).In addition,mRNA and protein levels of NNLRP3, ASC, Caspase-1, IL-1β and GSK-3β expressions in MI group were clearly increased (P<0.01) compared with those in sham group.These indicators were significantly decreased in SB+MI group (P<0.01). Interestingly, the indicators were all higher at 7 days than 2 days. CONCLUSION GSK-3β inhibition induces cardioprotection via attenuating the activation of NLRP3 inflammasome after the acute myocardial infarction in rats.
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OBJECTIVE To explore the effects of perinatal inflammation on the expression of proin-flammatory cytokines and DMEs (drug metabolism enzymes) in offspring mice. METHODS C57BL/6 maternal mice were administrated with single dose 100 μg·kg-1LPS(lipopolysaccharide)or saline(vehicle) during gestation (day 10 after fertilization). Offspring mice were sacrificed at 30 d after birth and liver samples were collected.Real-time PCR was adopted to test the mRNA expression of proinflammatory cytokines (Nrlp3 and IL-1β), nuclear receptors (Pxr and Car), and DMEs (Cyp3a11, 2b10, 1a2, and Ugt1a1).RESULTS Gender different expression of candidate genes was observed.The expression of Car,in the maternal injection of LPS groups,was significantly decreased in both female and male offspring (n=3-8/group, P<0.01). Concomitantly, a significantly lower expression of Cyp3a11 was found in both female and male offspring (P<0.01, P<0.05, respectively). Furthermore, the expression of Ugt1a1 was reduced in male offspring following maternal administration of LPS (P<0.01). In male offspring, Nrlp3 expression was specially decreased(P<0.05).Interestingly,there was an approximately 66% reduction in mRNA level of Cyp1a2 in female offspring (P<0.01), while in male offspring Cyp1a2 expression showed an increased trend (P>0.05) compared with vehicle group. The expression of Pxr, Cyp2b10, and IL-1β was no difference between LPS treatment group and vehicle group(P>0.05).CONCLUSION Maternal LPS administration affects the expression of proinflammatory cytokines, nuclear receptors and DMEs in mouse offspring.
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OBJECTIVE To demonstrate the long-or short-term impacts of neonatal Pxr(pregnane X receptor) agonists exposureon DMEs (drug metabolism enzymes) expression in adulthood. METHODS C57BL/6 mice(day 5,postnatal)were injected with different doses(0,50,100,150,200 mg·kg-1·d-1, constitutive 4 d)of PCN(pregnenolone-16a-carbonitrile).Mice at different ages(day 5,10,15,25,postna-tal)were administrated with 200 mg·kg-1·d-1PCN in constitutive 4 d.All mice were sacrificed at day 60 after birth. Liver samples were collected for detecting the expression of Pxr target genes. RESULTS Compared with vehicle group, the significant inductions of Cyp2b10, Cyp3a11 and Pxrwere observed in high dose groups (150, 200 mg·kg-1·d-1, 5-8 d after birth) both in male and female mice (n=4-9/group,P<0.05).Furthermore,high dose groups(200 mg·kg-1·d-1,5-8 d after birth)were found to have higher mRNA expression levels of Cyp2a4,Ugt1a1,Abcc4,and Oatpla4 in female mice,while Papss2 in male mice compared with vehicle groups (n= 4-9/group, P<0.05). Interestingly, a decreased mRNA expression of Sult2a1 was identified in 200 (5-8 d) groups (n=4-9/group, P<0.05). Consistent with these results, the protein expression of Cyp3a11 was only increased in 200 (5-8 d) groups compared with the vehicle groups(n=3/group,P<0.05).Importantly,the persistent impacts on DMEs only occurred in day 5 and day 25 treatment groups,not day 10 and day 15 groups(n=4/group).CONCLUSION Neonatal Pxr activation has a long-term effect on the expression of DMEs in C57BL/6 mice.Dose and treatment exposure time are two key factors involved in this permanent alteration procedure.
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This study was designed to investigate effects of pargyline on histone methylation in the promoter and enhancer regions and transcription of cytochrome P450 3A4/3A7 (CYP3A4/3A7) gene. Human primary fetal liver cells were isolated, cultured and randomly divided into several groups including control, solvent, pargyline low, middle, high dose (treated with 0.6, 1.2, 2.4 mmol·L-1). HepG2 cells were cultured and treated with 0.03, 0.3, 3 mmol·L-1 pargyline. After 48 hours, total RNAs were prepared from the cells to determine the expression of CYP3A mRNA in primary fetal cells and HepG2 cells with real-time quantative PCR (qPCR). HepG2 cells were cultured and then treated with 3 mmol·L-1 pargyline for 48 hours. The chromatin immunoprecipitation (ChIP) assay was performed with dimethylation of histone H3 at lysine 4 (H3K4me2), and IgG antibodies respectively. The precipitated DNA was resuspended and used for qPCR. Primers were used to detect different regions of CYP3A4/3A7 promoter and enhancer. Occupancy of H3K4me2 was shown as percent of input DNA relative to control cells. The results suggested that pargyline has an effect on primary fetal liver cells and HepG2 cells proliferation. The level of CYP3A7 was markedly enhanced in human primary fetal liver cells by treatment with 1.2, 2.4 mmol·L-1 of pargyline (P-1 of pargyline in HepG2 cells (P<0.001) compared with solvent control. Occupancy of H3K4me2 on human CYP3A4 promoter (-362 to +53) and enhancer segment (-7 836 to -6 093) harbored the overlapping hepatocyte nuclear factors 4A (HNF4A) binding site compared with a negative control. Occupancy of H3K4me2 on human CYP3A7 promoter (-163 to +103) and enhancer segment (-4 054 to -3 421, -6 265 to -6 247) overlapped with glucocorticoid receptor (GR) binding site. In conclusion, the enriched H3K4me2 in the promoter and enhancer regions was induced by pargyline with HNF4A or GR binding site in CYP3A4/3A7 gene to activate the corresponding genes.
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<p><b>OBJECTIVE</b>To investigate the effect of midazolam on human ether-a-go-go (hERG) K+ channels exogenously expressed in human embryonic kidney cells (HEK-293) and the underlying molecular mechanisms.</p><p><b>METHODS</b>Whole-cell patch clamp technique was used to record WT, Y652A and F656C hERG K+ current expressed in HEK-293 cells.</p><p><b>RESULTS</b>Midazolam inhibited hERG K+ current in a concentration-dependent manner, the half-maximum block concentrations (IC50) values were (1.31 ± 0.32) µmol/L. The half-activation voltage (V1/2) were (2.32 ± 0.38) mV for the control and (-1.96 ± 0.83) mV for 1.0 µmol/L midazolam. The half-inactivation voltage (V1/2) was slightly shifted towards negative voltages from (-49.25 ± 0.69) mV in control to (-57.53 ± 0.53) mV after 1.0 µmol/L midazolam (P < 0.05). Mutations in drug-binding sites (Y652A or F656C) of the hERG channel significantly attenuated the hERG current blockade by midazolam.</p><p><b>CONCLUSION</b>Midazolam can block hERG K+ channel and cause the speed of inactivation faster. Mutations in the drug-binding sites (Y652 or F656) of the hERG channel were found to attenuate hERG current blockage by midazolam.</p>
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Humans , Dose-Response Relationship, Drug , Ether-A-Go-Go Potassium Channels , HEK293 Cells , Midazolam , Pharmacology , Mutation , Patch-Clamp Techniques , Potassium Channel Blockers , Pharmacologyالملخص
<p><b>BACKGROUND</b>Quantitative magnetic resonance imaging (qMRI) of articular cartilage represents a powerful tool in osteoarthritis research, but has so far been confined to a field strength of 1.5 T. The aim of the study was to determine the reproducibility and accuracy of qMRI assessments of the knee cartilage volume by comparing quantitative swine cartilage volumes of the sagittal (sag) multi echo data imagine combination water-excitation (MEDICwe) sequence and the fast low-angle shoot water-excitation (FLASHwe) sequence at 3.0-T MRI to directly measured volumes (DMV) of the surgically removed articular cartilage.</p><p><b>METHODS</b>Test-retest MRI was acquired in 20 swine knees. Two sag FLASHwe sequences and two sag MEDICwe sequences (spatial resolution 0.4 mm × 0.4 mm × 1.0 mm of 3-dimension (3D) were acquired at 3-T MRI in a knee. Articular cartilage volume was calculated from 3D reformations of the MRI by using a manual program. Calculated volumes were compared with DMV of the surgically removed articular cartilage. Knee joint cartilage plates were quantified paired in order.</p><p><b>RESULTS</b>In the knee joint of swine, reproducibility errors (paired analysis) for cartilage volume were 2.5% to 3.2% with sag FLASHwe, and 1.6% to 3.0% with sag MEDICwe. Correlation coefficients between results obtained with qMRI and DMV ranged from 0.90 to 0.98 for cartilage volume. Systematic pairwise difference between results obtained with qMRI and DMV ranged from -1.1% to 2.8%. Random pairwise differences between results obtained with qMRI and DMV ranged from (2.9 ± 2.4)% to (6.8 ± 4.5)%.</p><p><b>CONCLUSIONS</b>FLASHwe and MEDICwe sequences permit highly accurate and reproducible analysis of cartilage volume in the knee joints of swine at 3-T MRI. Cartilage volume reproducibility for the MEDICwe data is slightly higher than the FLASHwe data.</p>
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Animals , Cartilage, Articular , Pathology , Magnetic Resonance Imaging , Methods , Swineالملخص
To understand the HA1 genetic variation characterization of influenza H3N2 virus isolates in Zhu-hai during 2008-2009, we selected 20 of H3N2 Influenza strains cultured in MDCK cell. Viral RNAs were extracted and amplified by using RT-PCR. The amplified products were purified after identified by gel electrophoresis and then the nucleotide sequences of the amplicons were determined. The results were analyzed by the software ClustalX and MEGA4. 1. When compared with the amino acid sequences of the epitopes of HA1 district of H3N2 influenza vaccine recommended by WHO in 2008, changes were found in those of H3N2 influenza strains in Zhuhai in 2008: K140I in all of H3N2 influenza strains, L157S in 08-0343 and 08-0677, K158R in 08-0466, 08-0620 and 08-0667, K173E in 08-0466 and 08-0620, K173N in 08-0667, and I192T in 08-0667. The epitopes of HA1 district of H3N2 influenza strains in Zhuhai in 2009 are different from that of H3N2 influenza vaccine during the same time: K173Q and P194L occur in all of H3N2 influenza strains, N144K, K158N, and N189K occur in the strains except the strain 09-0056. HA1 domain of H3N2 influenza strains in 2009 has lost a glycosylation site at amino acid position 144 while the glycosylation sites of HA1 domain of H3N2 influenza stains isolated in 2008 remained. This study suggested that H3N2 influenza virus in Zhuhai in 2008 was not evolved a novel variant and H3N2 influenza variant in 2009 was attributed to antigenic drift in HA1 district.
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Animals , Dogs , Humans , Antigens, Viral , Allergy and Immunology , Cell Line , China , Epitopes , Allergy and Immunology , Glycosylation , Hemagglutinin Glycoproteins, Influenza Virus , Chemistry , Genetics , Allergy and Immunology , Metabolism , Influenza A Virus, H3N2 Subtype , Classification , Genetics , Allergy and Immunology , Mutation , Phylogeny , Sequence Analysis, DNAالملخص
<p><b>OBJECTIVE</b>To observe the sensitivity of the PC-3 cell lines transfected with the PCI-NEO-SNCG plasmid to Cisplatin (DDP), 5-Fluorouracil (5-FU), Adriamycin (ADM), Vincristine (VCR) and Paclitaxel (TAX), and to explore the influence of the SNCG expression on the effectiveness of anti-tumor drugs.</p><p><b>METHODS</b>The plasmids PCI-NEO and PCI-NEO-SNCG were transfected into the hormone-independent prostate cancer cell lines PC-3. RT-PCR was adopted to examine the expression of SNCG in the PC-3 cell lines. The MTT method was employed to detect the suppressive effects of different anti-tumor drugs (DDP, ADM, 5-FU, VCR and TAX) on the cell lines transfected with PCI-NEO and PCI-NEO-SNCG. Flow cytometry was used to analyze the cell cycles and apoptosis of the transfected cells treated with TAX.</p><p><b>RESULTS</b>The 5 anti-tumor drugs suppressed the growth of the cell lines transfected with the plasmids PCI-NEO and PCI-NEO-SNCG in a time-dependant manner. The comparison between the growth-suppressing effects of different anti-tumor drugs on the PC-3 cell lines showed no significant differences between the group transfected with PCI-NEO and that with PCI-NEO-SNCG in DDP, 5-FU, ADM and VCR (P > 0.05), while the rate of suppression of TAX on the latter cell lines was significantly lower than that on the former (P < 0.01). Compared with the PCI-NEO-SNCG plasmid transfected cell lines, after treated with TAX for 48 hours, those transfected with the PCI-NEO plasmid exhibited a significantly larger proportion of cells remaining in the G2-M stage (P < 0.01), a smaller proportion in the G0-G1 and S stages (P < 0.01) and a significantly higher expression of Caspase-3 (P < 0.01).</p><p><b>CONCLUSION</b>The significant reduction of the growth-suppressing effect of TAX in the SNCG-transfected PC-3 cell lines suggests that the expression of SNCG may restrain the effect of TAX. These findings have provided evidence and guide to the individual chemotherapy of prostate cancer.</p>
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Humans , Male , Antineoplastic Agents , Pharmacology , Breast Neoplasms , Genetics , Cell Line, Tumor , Cisplatin , Pharmacology , Drug Screening Assays, Antitumor , Neoplasm Proteins , Genetics , Paclitaxel , Pharmacology , Prostatic Neoplasms , Transfection , gamma-Synuclein , Geneticsالملخص
Objective To study the neuroprotective effect of erythropoietin(EPO) on neonatal rats model with hypoxia-ischemia encephalopathy(HIE).Methods HIE was induced in rats on 7th day of postnatal age by ligation of right common carotid artery,followed by 2 h of hypoxia(80 mL/L O2).The subjects were divided into sham-operated group,control group and EPO group.EPO 4 000 U/(kg?day) was injected daily from day 2 pre-surgery for 9 to 16 days and PBS was injected in the control group.The neuroprotective effect of EPO on HIE model was detected by brain weight,the difference in weights between the ipsilateral(right) and contralateral(left) brain and the function test.In vitro study,the neural progenitor cell line C17.2 under gone apoptosis following an ischemia-like metabolic inhibition.The effect of EPO on the cell line ischemia modle 17.2 was evaluated by detecting Annexin V with flow cytometry.Results The signi-ficant and sustained brain injury in the hypoxia-ischemia and vehicle-treated group was observed and measured by reduction in relative weights of ipsilateral to contralateral and compromised sensorimotor functions in response to postural reflex test,compared with those of sham-operated animals(Pa