Impact of 1.8GHz Radiofrequency Electromagnetic Fields on Gene Expression Profiles in Rat Neurons / 预防医学

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.

Role of platelet in angiogenesis / 浙江大学学报·医学版

Angiogenesis is a process involving the growth of new blood vessels from pre-existing vessels though sprouting or other ways. It plays an important role in both physiological and pathological processes. Researches have found that platelets may contribute to angiogenesis as well. In this paper, we review the role of platelet in angiogenesis, especially the relationship with tumor angiogenesis, and discuss clinical implications of these findings.

Effect of 1.8 GHz radiofrequency electromagnetic fields on gene expression of rat neurons / 中华劳动卫生职业病杂志

<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>

Preliminary study on role of lipid rafts in receptor clustering induced by 50 Hz magnetic fields and its mechanism / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the relationship among a 50 Hz magnetic field (MF)-induced epidermal growth factor receptor (EGFR) clustering,lipid rafts and acid sphingomyelinase (ASM), and to explore its possible mechanism.</p><p><b>METHODS</b>Human amnion FL cells were exposed to 50 Hz, 0.4 mT MF for 15 min. EGF treatment was used as positive control. Nystatin was employed to study lipid rafts since it could disrupt lipid rafts structure.The EGF receptors, ASM and lipid rafts were labeled with polyclonal anti-EGFR antibody, anti-ASM antibody and FITC-Cholera toxin B, respectively. The images were observed by laser confocal scanning microscope.</p><p><b>RESULT</b>Both EGF treatment and 50 Hz MF exposure could induce EGFR clustering; however, nystatin pretreatment disrupted this effect. MF exposure turned ASM (labeled with Cy3) from a diffused state in the sham exposure group to a concentrated state on the cell membrane, which co-localized with lipid rafts (labeled with FITC).</p><p><b>CONCLUSION</b>The results suggest that the EGFR clustering induced by 50 Hz MF depends on intact lipid rafts on cellular membrane, and the ASM might participate in the process of EGFR clustering.</p>

Effects of 50 Hz magnetic fields on DNA double-strand breaks in human lens epithelial cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of 50 Hz magnetic fields (MF) on DNA double-strand breaks in human lens epithelial cells (hLECs).</p><p><b>METHODS</b>The cultured human lens epithelial cells were exposed to 0.4 mT 50 Hz MF for 2 h, 6 h, 12 h, 24 h and 48 h. Cells exposed to 4-nitroquinoline-1-oxide, a DNA damage agent, at a final concentration of 0.1 micromol/L for 1 h were used as positive controls.After exposure, cells were fixed with 4 % paraformaldehyde and for H2AX (gamma H2AX) immunofluorescence measurement. gamma H2AX foci were detected at least 200 cells for each sample. Cells were classified as positive when more than three foci per cell were observed. Mean values of foci per cell and percentage of foci positive cells were adopted as indexes of DNA double-strand breaks.</p><p><b>RESULT</b>The mean value of foci per cell and the percentage of gamma H2AX foci positive cells in 50 Hz MF exposure group for 24 h were (2.93 +/-0.43) and (27.88 +/-2.59)%, respectively, which were significantly higher than those of sham-exposure group [(1.77 +/-0.37) and (19.38+/-2.70)%, P <0.05], and the mean value of foci per cell and the percentage of gamma H2AX foci positive cells in 50 Hz MF exposure group for 48 h were (3.14 +/-0.35) and (31.00 +/-3.44)%, which were significantly higher than those of sham-exposure group (P <0.01). However there was no significant difference between 50 Hz MF exposure groups for 2 h, 6 h, 12 h and sham-exposure group for above two indexes (P >0.05).</p><p><b>CONCLUSION</b>0.4 mT 50 Hz MF exposure for longer duration might induce DNA double-strand breaks in human lens epithelial cells in vitro.</p>

Effects of millimeter wave on gene expression in human keratinocytes / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To explore the effect of millimeter wave exposure at low power density on gene expression in human keratinocytes (HaCaT).</p><p><b>METHODS</b>HaCaT keratinocytes were exposed to 30.16 GHz millimeter wave with power densities of 1.0 or 3.5 mW/cm2 for 30 min per day. Gene expression profiles were obtained using the Affymetrix human genome U95A GeneChip. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to confirm the differential expression of genes obtained from Genechip analysis.</p><p><b>RESULT</b>PAR-2 and ERGIC-53 genes in HaCaT cells were up-regulated by 3.5 mW/cm2 millimeter wave exposure for 4 times. ERGIC-53 gene was also up-regulated by 1.0 mW/cm2 millimeter wave exposure for 4 times. However, no significant change for PAR-2 expression was found after the same exposure.</p><p><b>CONCLUSION</b>Millimeter wave exposure could affect gene expression in human keratinocytes, which might be related to the intensity and the times of exposure.</p>

Noise magnetic fields mitigates the inhibition of secretion function of primary human villous trophoblasts induced by 50 Hz magnetic fields / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To explore the effects of 50 Hz sinusoidal magnetic fields (MF) on secretion function of primary human villous trophoblasts in vitro, and the interference effect of "noise" MF.</p><p><b>METHODS</b>The trophoblasts were isolated from human villus by trypsin digestion and incubated in DMEM medium.Then the trophoblasts were exposed to 0.4 mT 50 Hz MF and/or "noise" MF respectively for different durations. Each exposure group was matched with one control group which was from the same villus and cultured with the same condition except the MF exposure. The concentrations of human chorionic gonadotropin (HCG) and progesterone in the culture medium were measured by immunofluorescence. Statistical significance of differences between means was determined by one way-ANOVA with P<0.05 considered significant.</p><p><b>RESULT</b>50 Hz MF inhibited the HCG and progesterone secretion significantly when exposure for 72 h (compared with control group, P<0.05). There was no significant change of HCG and progesterone secretion when trophoblasts were exposed to 0.4 mT "noise" MF within 72 h (compared with control group, P>0.05). However, by superimposing the "noise" MF, the inhibition of HCG and progesterone secretion of trophoblasts induced by 50 Hz MF was eliminated.</p><p><b>CONCLUSION</b>The exposure to 50 Hz MF for long period could inhibit trophoblasts secreting HCG and progesterone, and the "noise" MF with the same intensity could eliminate the effects induced by 50 Hz MF.</p>

Renal protective activity of Hsian-tsao extracts in diabetic rats / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the renal protective activity of Hsian-tsao Mesona procumbens Hemsl. water extracts in diabetic rats.</p><p><b>METHODS</b>Thirty Sprague-dawley female rats were randomly divided into three groups (n = 10 each), "control group" with intraperitoneal saline injection, "diabetic group" with 60 mg of intraperitoneal streptozotocin injection per kg of body weight and "Hsian-tsao group" with intragastric administration of Hsian-tsao extraction everyday for 4 weeks after intraperitoneal streptozotocin injection. The body weight and blood sugar were measured before and after model induction in the three groups. Thrombospondin-1 (TSP-1) expressions in the kidney were monitored by immunohistochemistry. Kidney ultrastructural changes were also analyzed by using transmission electron microscopy.</p><p><b>RESULTS</b>Before diabetic model induction, there were no significant differences among the three groups in body weight and blood sugar. Four weeks after the induction of diabetes, the differences became statistically significant. Electron microscopy also revealed disruption of the foot processes of the podocytes and other damages in diabetic group. These damages were significantly less severe in Hsian-tsao group when compared with the diabetic group. TSP-1 expressions in the kidney were significantly increased in both the diabetic group and Hsian-tsao group, but it was relatively lower in Hsian-tsao group than in diabetic group.</p><p><b>CONCLUSION</b>Our results showed that Hsian-tsao treatment in the diabetic rats effectively prevented the pathological alterations in the kidney and decreased the TSP-1 expression. It was suggested that Hsian-tsao had protective effect on the kidneys of the diabetic rats.</p>

Effect of 1.8 GHz radiofrequency electromagnetic fields on the expression of microtubule associated protein 2 in rat neurons / 中华劳动卫生职业病杂志

<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>

Effects of GSM 1800 MHz radiofrequency electromagnetic fields on DNA damage in Chinese hamster lung cells / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effects of GSM 1800 MHz radiofrequency electromagnetic fields (RF EMF) on DNA damage in Chinese hamster lung (CHL) cells.</p><p><b>METHODS</b>The cells were intermittently exposed or sham-exposed to GSM 1800 MHz RF EMF (5 minutes on/10 minutes off) at a special absorption rate (SAR) of 3.0 W/kg for 1 hour or 24 hours. Meanwhile, cells exposed to 2-acetylaminofluorene, a DNA damage agent, at a final concentration of 20 mg/L for 2 hours were used as positive control. After exposure, cells were fixed by using 4% paraformaldehyde and processed for phosphorylated form of H2AX (gammaH2AX) immunofluorescence measurement. The primary antibody used for immunofluorescence was mouse monoclonal antibody against gammaH2AX and the secondary antibody was fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Nuclei were counterstained with 4, 6-diamidino-2-phenylindole (DAPI). The gammaH2AX foci and nuclei were visualized with an Olympus AX70 fluorescent microscope. Image Pro-Plus software was used to count the gammaH2AX foci in each cell. For each exposure condition, at least 50 cells were selected to detect gammaH2AX foci. Cells were classified as positive when more than five foci were detected. The percentage of gammaH2AX foci positive cells was adopted as the index of DNA damage.</p><p><b>RESULTS</b>The percentage of gammaH2AX foci positive cell of 1800 MHz RF EMF exposure for 24 hours (37.9 +/- 8.6)% or 2-acetylaminofluorene exposure (50.9 +/- 9.4)% was significantly higher compared with the sham-exposure (28.0 +/- 8.4)%. However, there was no significant difference between the sham-exposure and RF EMF exposure for 1 hour (31.8 +/- 8.7)%.</p><p><b>CONCLUSION</b>1800 MHz RF EMF (SAR, 3.0 W/kg) for 24 hours might induce DNA damage in CHL cells.</p>

Effects of GSM 1800 MHz radiofrequency electromagnetic fields on protein expression profile of human breast cancer cell MCF-7 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effects of GSM 1800 MHz radiofrequency electromagnetic fields (RF EMF) exposure on protein expression profile of human breast cancer cell line (MCF-7), as to exploring the possible effects on normal cell physiological function.</p><p><b>METHODS</b>MCF-7 cells were continuously or intermittently (5 minutes field on followed by 10 minutes off) exposed to RF EMF for different duration (1 hour, 3 hours, 6 hours, 12 hours, or 24 hours) at an average specific absorption rate (SAR) of 3.5 W/kg. The extracted proteins were separated by 2-dimensional electrophoresis and the protein-spot distribution of the silver-stained gels was analyzed by using PDQuest software 7.1. Each experiment was repeated three times.</p><p><b>RESULTS</b>On the average, around 1100 proteins were detected using pH 4 - 7 IPG strip. There were no differential proteins found under continuous exposure at SAR of 3.5 W/kg for 6 hours. Under other exposure conditions, we found various differentially expressed proteins in exposure groups as compared with the sham-exposed controls. Especially in 3 hours intermittent exposure and 12 hours continuous exposure, eighteen and seven differential proteins were detected, respectively. The categories and functions of these differentially expressed proteins were analyzed by searching of SWISS-PROT protein database, which suggested that these proteins should be related to the functions of biosynthesization, signal transduction, and DNA damage and repair.</p><p><b>CONCLUSIONS</b>Data indicated that the protein expression changes induced by RF radiation might depend on exposure duration and mode. Many biological processes might be affected by RF exposure.</p>

Global gene response to GSM 1800 MHz radiofrequency electromagnetic field in MCF-7 cells / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate whether GSM 1800 MHz radiofrequency electromagnetic field (RF EMF) can change the gene expression profile in MCF-7 cells and to screen RF EMF responsive genes.</p><p><b>METHODS</b>Subcultured MCF-7 cells were intermittently (5-minute fields on/10-minute fields off) exposed or sham-exposed to GSM 1800 MHz RF EMF, which was modulated by 217 Hz EMF, for 24 hours at an average specific absorption rate (SAR) of 2.0 W/kg or 3.5 W/kg. Immediately after RF EMF exposure or sham-exposure, total RNA was isolated from MCF-7 cells and then purified. Affymetrix Human Genome U133A Genechip was applied to examine the change of gene expression profile according to the manufacturer's instruction. Data was analyzed by Affymetrix Microarray Suite 5.0 (MAS 5.0) and Affymetrix Data Mining Tool 3.0 (DMT 3.0). Quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to validate the differentially expressed genes identified by Genechip analysis.</p><p><b>RESULTS</b>A small number of differential expression genes were found in each comparison after RF EMF exposure. Through reproducible and consistent analysis, no gene or five up-regulated genes were screened out after exposure to RF EMF at SAR of 2.0 W/kg or 3.5 W/kg, respectively. However, these five genes could not be further confirmed by RT-PCR.</p><p><b>CONCLUSION</b>The present study did not provide clear evidence that RF EMF exposure might distinctly change the gene expression profile in MCF-7 cells under current experimental conditions, implying that the exposure might not affect the MCF-7 cell physiology, or this cell line might be less sensitive to the RF EMF exposure.</p>

Noise magnetic fields block co-suppression effect induced by power frequency magnetic field and phorbol ester / 中华预防医学杂志

<p><b>OBJECTIVES</b>To explore intervention with electromagnetic noise for co-suppression effect on gap-junctional intercellular communication (GJIC) induced or strengthened by low intensity magnetic field with carcinogen 12-O-tetradecanoylphorbol-13-acetate (TPA).</p><p><b>METHODS</b>Fibroblast cells from NIH 3T3 mice were exposed to extremely low intensity magnetic field (MF) 0.2 mT, 0.2 mT + TPA or/and electromagnetic noise with the same intensity of MF for 24 h, and GJIC was determined using fluorescence recovery analysis after photobleaching (FRAP) with a laser-scanning confocal microscope (Leica, Germany).</p><p><b>RESULTS</b>GJIC function could be co-suppressed by MF of 0.2 mT with TPA, with fluorescence recovery of (23 +/- 11)%, lower than that in the control group [(46 +/- 19)%] and in the group with TPA only [(34 +/- 17) %] (P < 0.01), indicating 0.2 mT MF plus TPA could co-inhibit GJIC (P < 0.01). Superposition of 0.2 mT noise MF could get a fluorescence recovery of (35 +/- 19)% and significantly antagonize its co-suppression by TPA.</p><p><b>CONCLUSION</b>Electromagnetic noise of 0.2 mT could block the intensifying effect of power frequency magnetic field on TPA-induced GJIC inhibition.</p>