ABSTRACT
ObjectiveTo investigate the effects of hydroxycamptothecin liposomes （LHCPT） on myocardial fibrosis in rats with heart failure by regulating the sphingosine kinase 1 （SphK1）/sphingosine-1-phosphate （S1P） signaling pathway. MethodsSD rats were divided into control group， model group， hydroxycamptothecin （HCPT） group， LHCPT group， captopril group， and LHCPT+K6PC-5 （SphK1 activator） group， with 12 rats in each group. The heart failure rat models in all groups except the control group were established by intraperitoneal injection of doxorubicin and then the corresponding drugs were given once a day. After four weeks， we applied color Doppler ultrasound to detect left ventricular end systolic diameter （LVESD）， left ventricular end diastolic diameter （LVEDD）， and left ventricular ejection fraction （LVEF） in rats； HE and Masson staining for myocardial pathological damage and myocardial fibrosis in rats， respectively； ELISA method for the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in rat myocardial tissues； qRT-PCR for the expression of transforming growth factor-β1 （TGF-β1）， type I collagen （Collagen I）， and type Ⅲ collagen （Collagen Ⅲ） in rat myocardial tissues； Western blot for the expression of SphK1 and S1P proteins in rat myocardial tissues. ResultsCompared with the control group， the model group showed severe myocardial pathological damage and myocardial fibrosis， increased LVESD， LVEDD， levels of TNF-α and IL-6， expression of TGF-β1， Collagen I， Collagen Ⅲ， SphK1， S1P and decreased LVEF （P<0.05）. Compared with the model group， the HCPT group， LHCPT group and captopril group showed alleviated myocardial pathological damage and myocardial fibrosis， decreased LVESD， LVEDD， levels of TNF-α and IL-6， expression of TGF-β1， Collagen I， Collagen Ⅲ， SphK1， S1P and increased LVEF （P<0.05）. Compared with the LHCPT group， the LHCPT+K6PC-5 group showed aggravated myocardial pathological damage and myocardial fibrosis， increased LVESD， LVEDD， levels of TNF-α and IL-6， expression of TGF-β1， Collagen I， Collagen Ⅲ， SphK1， S1P and decreased LVEF （P<0.05）. ConclusionLHCPT may inhibit myocardial fibrosis in heart failure rats by inhibiting the SphK1/S1P signaling pathway.
ABSTRACT
ObjectiveTo evaluate the association between Pro12Ala polymorphism in peroxisome proliferator activated receptorγ2 (PPARγ2) gene and gestational diabetes mellitus(GDM).Methods Publications on genetic association studies of PPARγ2 and GDM were searched using the PubMed database, The HuGE Navigator, China National Knowledge Infrastructure (CNKI), Wanfang database and VIP Science from the inception of the databases to December 1, 2014. Two reviewers independently selected literature according to the inclusion and exclusion criteria, extracted data and assessed the quality of the data using the Newcastle-Ottawa Scale (NOS) standard. Meta-analysis was performed using RevMan 5.3 software.ResultsOverall, 13 eligible articles were identified, including seven in English and six in Chinese, with a total of 2 787 GDM cases and 5 408 healthy controls. Quality assessment showed that the quality of the 13 articles was all good, with NOS≥5. (1) Pro12Ala polymorphism in PPARγ2 (allele Ala or genotype Ala/Ala or Pro/Ala) was shown to be highly associated with GDM occurrence on general evaluation, with anOR(95%CI) of 0.74(0.60-0.93) in the allele model and 0.79(0.65-0.96) in the dominant genetic model (P0.05, respectively), although there was still a significant correlation in polymerase chain reaction-restriction fragment length polymorphism with anOR(95%CI) of 0.58(0.43-0.79) in the allele model and 0.62(0.45-0.85) in the dominant genetic model (P<0.01, respectively).ConclusionsThe Ala allele and the Ala/Ala or Pro/Ala genotypes of the Pro12Ala polymorphism in PPARγ2 can decrease the risk of GDM. However, there are differences in the results which are affected by the genotype analysis method or races.
ABSTRACT
We found that the number of institutions made use of the undocumented in vitro diagnostic reagent in the survey. The phenomenon poses some risks and problems. In use this paper, we analyzed the situation and the reasons for the use of the undocumented in vitro diagnostic reagents, and put forward the corresponding measures.
Subject(s)
Humans , Indicators and Reagents , Reference StandardsABSTRACT
Objective To investigate the changes of gene expression in rat neurons induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) and to screen RF EMF-responsive genes. Methods The rat primary cultured neuronal cells were divided into two groups, the radiation group and control group, from which the total RNA was extracted immediately and purified after intermittently (5min on/10min off) exposed or U34 array was applied to detect the changes of gene expression in rat neurons. Results Among 1200 candidate genes, 24 up-regulated and 10 down-regulated genes which are associated with multiple cellular functions (cytoskeleton, signal transduction pathway, metabolism, etc.) after functional classification were found by using Affymetrix microarray suite software 5.0. Although the changes in gene expression were less than 2 folds, they had statistical significance (P<0.01). Conclusion RF radiation of 1.8GHz induce the changes of many genes transcription in rat neurons, some of which indicate the negative effects of RF radiation on neurons.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the changes of gene expression in rat neuron induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) to screen for RF EMF-responsive genes and the effect of different exposure times and modes on the gene expression in neuron.</p><p><b>METHODS</b>Total RNA was extracted immediately and purified from the primary culture of neurons after intermittent exposed or sham-exposed to a frequency of 1.8 GHz RF EMF for 24 hours at an average special absorption rate (SAR) of 2 W/kg. Affymetrix Rat Neurobiology U34 array was applied to investigate the changes of gene expression in rat neuron. Differentially expressed genes (Egr-1, Mbp and Plp) were further confirmed by semi-quantitative revere transcription polymerase chain reaction (RT PCR). The expression levels of Egr-1, Mbp and Plp were observed at different exposure times (6, 24 h) and modes (intermittent and continuous exposure).</p><p><b>RESULTS</b>Among 1200 candidate genes, 24 up-regulated and 10 down-regulated genes were found by using Affymetrix microarray suite software 5.0 which are associated with multiple cellular functions (cytoskeleton, signal transduction pathway, metabolism, etc.) after functional classification. Under 24 h and 6 h intermittent exposure, Egr-1 and Plp in experiment groups showed statistic significance (P < 0.05) compared with the control groups, while expression of Mbp did not change significantly (P > 0.05). After 24 h continuous exposure, Egr-1 and Mbp in experiment groups showed statistic significance (P < 0.05) compared with the control group, while expression of Plp did not change significantly (P > 0.05). Under the same exposure mode 6 h, expression of all the 3 genes did not change significantly. Different times (6, 24 h) and modes (intermittent and continuous exposure) of exposure exerted remarkable different influences on the expression of Egr-1, Mbp, Plp genes (P < 0.01).</p><p><b>CONCLUSION</b>The changes of many genes transcription were involved in the effect of 1.8 GHz RF EMF on rat neurons; Down-regulation of Egr-1 and up-regulation of Mbp, Plp indicated the negative effects of RF EMF on neurons; The effect of RF intermittent exposure on gene expression was more obvious than that of continuous exposure; The effect of 24 h RF exposure (both intermittent and continuous) on gene expression was more obvious than that of 6 h (both intermittent and continuous).</p>
Subject(s)
Animals , Rats , Cells, Cultured , Dose-Response Relationship, Radiation , Down-Regulation , Radiation Effects , Electromagnetic Fields , Neurons , Metabolism , Radiation Effects , Up-Regulation , Radiation EffectsABSTRACT
<p><b>OBJECTIVE</b>To explore the differential proteomic expression in human liver cells L-02 after exposure to HQ.</p><p><b>METHODS</b>Subcultured L-02 cells were treated by HQ for 24 h at a 1 x 10(-4) mol/L concentration and a blank group was set as the control. Immediately after the treatment, total cellular proteins were extracted and separated by 2-DE, and the images were analyzed by PDQuest software. The experiment was totally repeated 3 times with 3 repetitions for each group every time. The well repeated spots were identified by MALDI-TOF MS and then searched in NCBI human protein database with Mascot.</p><p><b>RESULTS</b>About 1,000 spots per gel were found. Compared with the control group, 17, 18 and 24 spots were significantly altered in 3 separate experiments. The 4 well repeated spots were identified by MALDI-TOF MS as Rho GDP dissociation inhibitor GDI alpha, 6-phosphogluconolactonase, erbB3 binding protein EBP1 and lamin A/C, isoform 1 precursor. They were involved in cell skeleton, signal transduction and energy metabolization in functional classification.</p><p><b>CONCLUSION</b>Hydroquinone can change the protein expression in liver cells, which provides clues for exploring the toxic mechanism.</p>
Subject(s)
Humans , Cell Line , Electrophoresis, Gel, Two-Dimensional , Hepatocytes , Metabolism , Hydroquinones , Toxicity , Mass Spectrometry , Proteomics , Reproducibility of ResultsABSTRACT
<p><b>OBJECTIVE</b>To investigate the changes of gene expression in rat neurons induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) and to screen for the RF EMF-responsive genes.</p><p><b>METHODS</b>Newly-born SD rats in 24 hours were sacrificed to obtain cortex and hippocampus neurons. The cells were divided randomly into two groups: the experiment group (the irradiation group) and the control group (the false irradiation group). In the irradiation group, after twelve days' culture, neurons were exposed to 1.8 GHz RF EMF modulated by 217 Hz at a specific absorption rate (SAR) of 2 W/kg for 24 hours (5 minutes on/10 minutes off) while in the false control group, the neurons were put in the same waveguide as in the irradiation group, but were not exposed to any irradiation. The total RNA was isolated and purified immediately after exposure. The affymetrix rat neurobiology U34 assay was used for detecting the changes in gene expression profile according to the manufacturer's instruction. RF EMF-responsive candidate gene was confirmed by using ribonuclease protection assay (RPA).</p><p><b>RESULTS</b>Among 1200 candidate genes, the expression levels of 34 genes were up or down regulated. Microtubule associated protein 2 (Map2) gene was selected as the candidate and subjected to further analysis. RPA data clearly revealed that Map2 was statistically significantly up-regulated after neurons were exposed to the RF EMF (P < 0.05).</p><p><b>CONCLUSION</b>The modulation of gene expression and function of Map2 as a neuron specific cytoskeleton protein is crucial to maintain the normal framework and function of neurons. The finding that 1.8 GHz RF EMF exposure increases the expression of Map2 might indicate some unknown effects of RF EMF on neurons.</p>
Subject(s)
Animals , Female , Male , Rats , Animals, Newborn , Cell Phone , Cells, Cultured , Dose-Response Relationship, Radiation , Down-Regulation , Electromagnetic Fields , Gene Expression , Radiation Effects , Microtubule-Associated Proteins , Genetics , Neurons , Metabolism , Radiation Effects , Radio Waves , Random Allocation , Rats, Sprague-Dawley , Up-RegulationABSTRACT
<p><b>OBJECTIVE</b>To investigate the toxic and DNA damaging effect of acrylamide (AA) on human keratinocytes and its mechanism.</p><p><b>METHODS</b>(1) After the keratinocyte cell line HaCaT cells were exposed to AA with different concentrations for 44 hours, cell survival rate was detected by MTT method. (2) The effects of DNA damage of exposed cells were detected by comet assay. (3) After treating the cells with 2.00 mmol/L of AA plus 0.50 mmol/L of 1-aminobenzotriazole (1-ABT), an inhibitor of cytochrome P-450 enzymes (CYP-450), for 4 hours, the relationship between DNA damage and CYP-450 was studied.</p><p><b>RESULTS</b>(1) Cytotoxicity measurement of AA showed that cell survival rate decreased significantly after 44-hour treatment. (2) Cytotoxicity was not detected after 4-hour AA treatment, but significant DNA damage was observed in all treatment groups, and the degree of damage increased with the concentration of AA. Moreover, the tail lengths of comet cells were in dose-effect relationship. As for cells treated by 1-ABT with 2 mmol/L AA, comet rate and tail length were 15.4% and (8.2 +/- 2.0) micro m respectively, which were decreased significantly (P < 0.01) when compared with 2 mmol/L AA treatment group [80.6% and (44.3 +/- 4.0) micro m].</p><p><b>CONCLUSIONS</b>Acrylamide has significant cytotoxicity and genotoxicity on HaCaT cells. AA-induced DNA damage may be related to the oxidative metabolite(s) of AA through CYP-450.</p>