Hui Wu1*, Dewen Wang2*, Yuhong Meng1, Hui Wang2, Yifan Xie2, Yang Li2, Hongyan Zuo2, Li Zhao2, Cuifang Zhang2 and Ruiyun Peng2 Author Affiliations 1Department of Pathology, Sixth Medical Center of PLA General Hospital, 100048, China 2Beijing Institute of Radiation Medicine. Beijing, China, 100850, China Received: April 04, 2022 | Published: June 03, 2022 Corresponding author: Hui Wu, Department of Pathology, the Sixth Medical Center of PLA General Hospital, 6 Fucheng roads, Beijing 100048, PR China Dewen Wang, Beijing Institute of Radiation Medicine, 27 Taiping roads, Beijing 100850, PR China DOI: 10.26717/BJSTR.2022.44.007029

In this study, rat pheochromocytoma (PC12) cells were induced into an Alzheimer's Disease (AD) neuronal model using nerve growth factor (NGF; 50 ng/mL) and Amyloid P25-35 (20 umol/L). Changes in the morphological structure, cell viability, apoptosis rate, and expression of apoptosis-related protein induced by exposure to a power frequency electromagnetic field (PF-MF; 50 Hz, 100 tiT, 24 h) were detected respectively by light and electron microscopy, the MTT assay, immunohistochemistry, flow cytometry and enzyme-linked immunosorbent assays. The results showed that 3-12 h after PF-MF exposure, the pathological injury was improved partly; metabolic activity was promoted and cell apoptosis was inhibited in the AD neuronal model. In addition, PF-MF exposure significantly inhibited the expression of Caspase8, Caspase3, and CytC, but increased the Bcl-2/Bax ratio of the AD neuronal model. Meanwhile, PF-MF seemed to have no effect on the expression of Fas and TNFR1. This study indicated that the mitochondria/caspase-dependent apoptotic pathway plays an important role in the positive effects of PF-MF on an AD neuronal model. The results suggested that PF-MF exposure might have potential therapeutic value for AD, and the underling molecular mechanisms still need further studies.

In a previous study, we reported the positive effects of extremely low frequency electromagnetic field (ELF-MF) exposure on Alzheimer's disease (AD) rats; however, the underlying mechanism remains unclear. In addition, we found that Raf-1 kinase inhibitor protein (RKIP) was downregulated by microwave exposure in the rat hippocampus. Our hypothesis was that RKIP-mediated NF-κB pathway signaling is involved in the effect of ELF-MF on the AD rat. In this study, D-galactose intraperitoneal (50 mg/kg/d for 42 d) and Aβ25-35 hippocampal (5 μL/unilateral, bilateral, single-dose) injection were implemented to establish an AD rat model. Animals were exposed to 50 Hz and 400 µT ELF-MF for 60 continuous days. The spatial memory ability of the rat was then tested using the Morris water maze. Protein expression and interaction were detected by western blotting and co-immunoprecipitation for RKIP-mediated NF-κB pathway factors. The results showed that ELF-MF exposure partially improved the cognitive disorder, upregulated the levels of RKIP, TAK1, and the RKIP/TAK1 interaction, but downregulated p-IKK levels in AD rats. These results indicated that RKIP-mediated NF-κB pathway signaling plays an important role in the ELF-MF exposure-mediated improvements in the AD rat. Our study suggested that ELF-MF exposure might have a potential therapeutic value for AD. Further in depth studies are required in the future.

Microwave radiation could increase the expression of pro-inflammatory cytokines in rat Sertoli cells, which may impair spermatogenesis. However, the mechanisms that microwave radiation induces the cytokine expression in Sertoli cells remain to be clarified. The activation of TLRs by their ligands can trigger a common signalling pathway to upregulate inflammatory cytokines such as IL-1, IL-6, IL-12 and TNF-α. Microwave radiation can increase the expression of TLRs in lymphocytes. The purpose of this study was to determine the effect of microwave radiation on the TLRs in rat testis. We focus on the effect of TLR2-5 (which is expressed relatively highly) by microwave radiation. The results showed that the expression of TLR2-5 and the pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) was increased both in mRNA and in protein. Furthermore, p-p38, p-ERK1/2, p-JNK and p-NF-κB p65, the key factors of TLR signalling, were also elevated by microwave exposure. And the NF-κB can be induced more dominantly. These results suggest that TLRs signalling can be activated by microwave radiation in testis, which may provide the molecular basis for the in-depth study.

The barbed nail characteristics,combat boot characteristics,biological contrastive test and hominine foot equivalent research are carried out for stabbing effect test and assessment. An equivalent hominine foot target is developed,of which soft tissues and bone structure are made of silicone rubber and epoxy. The target is made according to the standard man dimension and booted with a pair of special homebred combat boots. A stabbing effect evaluation model was established by using the stabbing position,depth,quantity and remaining things of the barbed nails stabbing into hominine foot as the input data based on the biological test,equivalent test and war injuries data. In the model,the degree of impact on operations is divided into levels 1- 4 from low to high. The effect of the barbed nail on personnel mobility could be assessed by using the effect assessment model and the test data of the equivalent targets.

In the present study, we investigated whether Raf-1 kinase inhibitory protein (RKIP) is important for neural cell apoptosis induced by microwave exposure and explored the role of MEK/ERK/CREB pathway regulated by RKIP in the apoptosis. Differentiated PC12 cells were exposed to continuous microwave radiation at 2.856 GHz for 5 min with average power density of 30 mW/cm2. RKIP sense and anti-sense recombinant plasmids were constructed and transfected into PC12 cells, respectively. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining and caspase-3 activity assay were used to detect cell apoptosis. The results showed that RKIP was downregulated after microwave exposure while the MEK/ERK/CREB signaling pathway was activated excessively. Moreover, the ratio of Bcl-2/Bax decreased, activity of caspase-3 increased, and thus apoptotic DNA fragmentation increased. RKIP overexpression significantly inhibited the phosphorylation of MEK, ERK, and CREB, while RKIP downregulation had the reverse effect. Furthermore, U0126 was found to antagonize the changes caused by RKIP downregulation after exposure to radiation. In conclusion, RKIP plays an important role in the neural cell apoptosis induced by microwave radiation, and the regulation of cell apoptosis by RKIP is partly through the MEK/ERK/CREB pathway. This suggests that RKIP may act as a key regulator of neuronal damage caused by microwave radiation.

Although some epidemiological investigations showed a potential association between long-term exposure of extremely low frequency electromagnetic fields (ELF-EMF) and Alzheimer's disease (AD), no reasonable mechanism can explain this association, and the related animal experiments are rare. In this study, ELF-EMF exposure (50 Hz 400 µT 60 d) combined with D-galactose intraperitoneal (50 mg/kg, q.d., 42 d) and Aβ25-35 hippocampal (5 μl/unilateral, bilateral, single-dose) injection was implemented to establish a complex rat model. Then the effects of ELF-EMF exposure on AD development was studied by using the Morris water maze, pathological analysis, and comparative proteomics. The results showed that ELF-EMF exposure delayed the weight gain of rats, and partially improved cognitive and clinicopathologic symptoms of AD rats. The differential proteomic analysis results suggest that synaptic transmission, oxidative stress, protein degradation, energy metabolism, Tau aggregation, and inflammation involved in the effects mentioned above. Therefore, our findings indicate that certain conditions of ELF-EMF exposure could delay the development of AD in rats.

To determine whether microwave (MW) radiation induces neural cell apoptosis, differentiated PC12 cells and Wistar rats were exposed to 2.856GHz for 5min and 15min, respectively, at an average power density of 30 mW/cm(2). JC-1 and TUNEL staining detected significant apoptotic events, such as the loss of mitochondria membrane potential and DNA fragmentation, respectively. Transmission electron microscopy and Hoechst staining were used to observe chromatin ultrastructure and apoptotic body formation. Annexin V-FITC/PI double staining was used to quantify the level of apoptosis. The expressions of Bax, Bcl-2, cytochrome c, cleaved caspase-3 and PARP were examined by immunoblotting or immunocytochemistry. Caspase-3 activity was measured using an enzyme-linked immunosorbent assay. The results showed chromatin condensation and apoptotic body formation in neural cells 6h after microwave exposure. Moreover, the mitochondria membrane potential decreased, DNA fragmentation increased, leading to an increase in the apoptotic cell percentage. Furthermore, the ratio of Bax/Bcl-2, expression of cytochrome c, cleaved caspase-3 and PARP all increased. In conclusion, microwave radiation induced neural cell apoptosis via the classical mitochondria-dependent caspase-3 pathway. This study may provide the experimental basis for further investigation of the mechanism of the neurological effects induced by microwave radiation.

Terrorism is a global issue and a constant international threat. As a result, anti-terrorism and emergency response strategies are tasks of critical importance that have a direct impact on the national security of every country in the world. This paper reviews new characteristics of international anti-terrorism measures and offers an in-depth reflection on emergency medical response countermeasures; additionally, this paper presents the goals of related research, which include: 1) to present a model of a highly efficient medical response command; 2) to introduce the pre-planning phases of the emergency medical response; 3) to establish a response system capable of handling various types of terror attacks; 4) to promote anti-terrorism awareness to the general public and emphasize its prevention; and 5) to continue basic investigations into emergency medical responses for various types of terrorist attacks（for example, the classifications and characteristics of new injuries, pathophysiology, prevention and treatment of the resultant stress disorders, improved high-efficiency medical response measures and equipment, etc.）.

OBJECTIVE:To study the effects of electromagnetic pulse (EMP), S-band high power microwave (S-HPM), and X-band high power microwave (X-HPM) on the Ca(2+) concentration and caspase-3 expression in Raji cells and the relationship between Ca(2+) concentration and caspase-3 expression, and to investigate the regulatory mechanism of electromagnetic radiation damage.METHODS:Raji cells were cultured conventionally. Some cells were irradiated by EMP, S-HPM, and X-HPM in the logarithmic growth phase for 6 hours and then collected; others received sham irradiation as a control. The Ca(2+) concentration in the cells was measured by laser scanning confocal microscope; the caspase-3 expression in the cells was evaluated by Western blot.RESULTS:Compared with the control group (Ca(2+) fluorescence intensity = 43.08 ± 2.08; caspase-3 expression level = 0.444 ± 0.13), the EMP,S-HPM, and X-HPM groups had significantly increased Ca(2+) concentrations, with Ca(2+) fluorescence intensities of 69.56 ± 1.71, 50.06 ± 1.89, and 70.68 ± 1.59, respectively (P < 0.01), and had upregulated caspase-3 expression, with expression levels of 0.964 ± 0.12, 0.586 ± 0.16, and 0.970 ± 0.07, respectively (P < 0.01). Each of the EMP and X-HPM groups had significantly higher Ca(2+) fluorescence intensity and caspase-3 expression level than the S-HPM group (P < 0.01), but there were no significant differences between the EMP and X-HPM groups. The linear regression analysis showed that the caspase-3 expression was upregulated as the Ca(2+) concentration increased, with a positive correlation between them (P < 0.01).CONCLUSION:EMP, S-HPM, and X-HPM cause damage probably by increasing the Ca(2+) concentration in cells and in turn inducing caspase-3 overexpression.

Ethnopharmacological relevance: The decoction of the flowers of Abelmoschus manihot (L.) Medic is traditionally used for the treatment of jaundice and various types of chronic and acute hepatitis in Anhui and Jiangsu Provinces of China. Phytochemical studies have indicated that total flavonoids extracted from flowers of Abelmoschus manihot (L.) Medic (TFA) were the major constituents of the flowers. The present study was designed to investigate the hepatoprotective effect of the plant extracts against carbon tetrachloride (CCl4) induced hepatocyte damage in vitro and liver injury in vivo.Material and methods: In the in vitro studies, freshly isolated rat hepatocytes were exposed to CCl4 (1%) along with/without various concentrations of TFA (4.5-72 mg/L). Cell damage was assessed by the trypan blue exclusion method and alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) in the medium were analyzed. In the in vivo studies, the hepatoprotective activity of TFA (125, 250 and 500 mg/kg) were investigated on CCl4-induced liver damages in mice. The levels of ALT, AST and ALP, gamma glutamyltransferase (gamma-GT), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) and nitric oxide (NO) were determined in serum. Hepatic malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and glutathione transferase (GST) were measured in the liver homogenates. Cytokine transcript levels of TNF-alpha, IL-1 beta and inducible nitric oxide synthase (iNOS) in the liver tissues of mice were measured using reverse transcription-polymerase chain reaction (RT-PCR). Livers were dissected out and evaluated for histomorphological changes.Results: A concentration-dependent increase in the percentage viability was observed when CCl4-exposed hepatocytes were treated with different concentrations of TFA. Levels of ALT, AST and ALP in the medium were significantly decreased. In the animal studies, TFA showed significant protection with the depletion of ALT, AST, ALP and gamma-GT in serum as was raised by the induction of CCl4. Moreover, TFA decreased the MDA level and elevated the content of GSH in the liver as compared to those in the CCl4 group. Furthermore, activities of antioxidative enzymes, including SOD, GPx, CAT and GST, were enhanced dose dependently with TFA. Meanwhile, the inflammatory mediators (e.g., TNF-alpha, IL-1 beta and NO) were inhibited by TFA treatment both at the serum and mRNA levels. Additionally, histological analyses also showed that TFA reduced the extent of liver lesions induced by CCl4.Conclusion: These results suggested that TFA protected mice against CCl4-induced liver injury through antioxidant stress and antiinflammatory effects. This finding justified the use of this plant in traditional medicine for the treatment of liver disease. (C) 2013 Elsevier Ireland Ltd. All rights reserved.

In order to define the neurobiological effects of extremely low frequency electromagnetic fields(ELF-EMFs),the cognitive function and hippocampal morphology of rats exposed to ELF-EMFs were experimentally investigated.Some male Wistar rats were exposed to ELF-EMFs(50 Hz,400 μT,1 V/m) for 60 d firstly,meanwhile these body weights were recorded weekly.After the exposure,the cognitive function,hippocampal morphology and content of Nissl bodies were measured by Morris water maze test,light/electronic microscopy and toluidine blue staining image analysis,respectively.The results showed that,during the exposure,the growth rate of rat body weight significantly decreased,but it restored after stopping exposure for 2 weeks.Immediately after the exposure being stopped,the average escape latency was significantly prolonged(P0.01),but it restored 7 d later.Part of the rat hippocampal neurons showed characteristic morphological changes of degeneration and apoptosis,and damages in synaptic structural,and the content of Nissl bodies reduced(P0.05);however,the lesions mentioned above recovered 15 d later and even restored 30 d later.It is concluded that the ELF-EMFs exposure(50 Hz,400 μT,60 d) can decrease the growth rate of body weight,cause cognitive dysfunctions,damage hippocampal formations,and reduce the content of Nissl bodies.However,these effects occur mainly in the early stage after the exposure being stopped,thus they were characterized by short duration and recoverability.

OBJECTIVE:To observe the toxicity of hyperoside in rat embryo-fetal development, in order to provide preference for safe use of drugs during gestation period.METHOD:Healthy pregnant rats were randomly divided into hyperosid groups (30, 175, 1000 mg x kg(-1) x d(-1)), the positive control group (cyclophosphamide, 7 mg x kg(-1) x d(-1)) and the solvent control group (1% aqueous carboxymethylcellulose). These rats were orally administered with hyperosid or vehicle during 6-15 d after gestation and subcutaneously injected with cyclophosphamide during 11-13 d. Maternal clinical sign, abortions, premature deliveries and body weight were monitored throughout gestation. At termination (gestation days 20), pregnant females were evaluated for clinical symposiums, weight change, corpora lutea count, existence and death of embryos; live fetuses were examined for gender, external, visceral and skeletal malformation and variations.RESULT:All pregnant rats showed no significant abnormality in appearance, viscera and skeletal development. However, there was a difference between the high-dose group of hyperoside and negative control group in the fetus body weight, the length of the embryos and the length of tail (P < 0.05).CONCLUSION:Pregnant women are suggested to cautiously use hyperoside because it shows certain impact on development of fetal rats under the experimental conditions.

Objective: To investigate the chronic toxicity of hyperoside in rats for evaluating its safety and providing a recommendation in defining its clinical dose.Methods: Wistar rats(n=160) were randomly divided into four groups of 40 animals each(20 per sex).They were administered with hyperoside 30,175 and 1 000 mg·kg-1·d-1 by gavage for 6 months followed by subsequent 1 month recovery period;1% aqueous carboxymethylcellulose was used as the control.Mortality,clinical signs,body weight changes,food consumption,hematological and biochemical parameters,gross findings,organ weights and histological markers were monitored during the dosing and recovery periods.Results: Compared with normal control group,no significant differences were observed in rats' behaviors,food-intake,weight gain and related examination indexes among the treatment groups.Hematology index,blood biochemical index,viscera coefficient and histological examination indicated that kidney was possible toxic target organs but the damage was reversible.Conclusion: In general,hyperoside has a better safety.Kidney is possible toxic target organs,but the damage is reversible and can return to normal after withdrawal.We suggest that regular hematology and kidney function index monitoring are necessary when hyperoside is clinically used.

To further assess hyperoside as a potential new anti-hepatitis B virus (HBV) drug, the safety of hyperoside extracted from Abelmoschus manihot (L.) Medic was evaluated by testing its acute toxicity and mutagenic risk. To test the acute toxicity of hyperoside, we determined the median lethal dose (LD 50 ) in mice. Forty healthy BALB/c mice (20 per sex) were administered a single oral dose of 5000 mg/kg hyperoside via the intragastrical route. The number of animals poisoned and died was noted daily for 14 consecutive days. All animals survived and appeared active and normal, indicating that the LD 50 of hyperoside was more than 5000 mg/kg. Potential genotoxicity of hyperoside was investigated using a bacterial reverse mutation assay (Ames test), a chromosome aberration test in Chinese hamster lung (CHL) fibroblasts, and an in vivo micronucleus test in rat bone marrow cells. In the bacterial reverse mutation assay, we observed no increases in the number of revertant colonies at any concentrations of hyperoside regardless of metabolic activation (S9) in all tester strains (TA97, TA98, TA100 and TA102) compared to the vehicle control (P>0.05). Hyperoside did not cause significant structural aberration in CHL cells in the presence or absence of S9 (P>0.05). The micronuclei rates of mice bone marrow cell in all groups showed no significant difference when compared with the negative control (P>0.05). In summary, hyperoside showed no genotoxicity in our experimental conditions.

Objective To explore the changes of SOX9 and WT1 expressions in rat Sertoli cells irradiated by EMP ( electromagnetic pulse),S-HPM ( S-band high power microwave) and X-HPM ( Xband high power microwave).Methods Primary Sertoli cells were isolated from 3-week-old Wistar rats and its purity was immunocytochemistrically indentified with WT1.After exposure to 6 × 104 V/m EMP,100 mW/cm2 S-HPM and X-HPM for 4 min respectively,SOX9 and WT1 expressions in Sertoli cells were determined with real-time PCR and Western blot,respectively.Results SOX9 mRNA expression was decreased at 6 and 12 h post-irradiation of three different bands of electromagnetic microwave ( F =15.20and 4.84,P ＜ 0.05 ).SOX9 protein expression was also decreased at 6 and 24 h after irradiation ( F =8.46 and 7.47,P＜0.05).WT1 mRNA expression was decreased at6 and 12 h (F=13.46 and 5.08,P ＜ 0.05 ),but its protein expression was decreased only at 24 h post-irradiation ( F =10.26,P ＜ 0.05 ).Conclusions Three bands of electromagnetic radiation reduce the expressions of SOX9 and WT1 in rat Sertoli cells,which may provide molecular foundation for genital system hazards induced by microwave radiation.

AIM:To construct the eukaryotic expression vectors of RKIP plasmid and detect its expression in PC12 cells.METHODS:The coding sequence of RKIP was generated by nested-PCR using total RNA extracted from the root ganglion neurons of rats. RKIP gene was cloned into the eukaryotic expression vector pcDNA3.0. After restriction enzyme analysis and sequence identification, the recombinant plasmid was transfected into PC12 cells with non-liposome mediated method by Vigofect. The expression of RKIP was detected by Western blot.RESULTS:The results of enzyme analysis and sequencing both identified DNA sequence of recombinant plasmid pcDNA3.0-RKIP correctly. The expression of RKIP increased obviously after transfection into PC12 cells.CONCLUSION:The eukaryotic expression plasmid of pcDNA3.0-RKIP was constructed successfully and it can be sustainly expressed in PC12 cells. This provides experimental basis for further study on the neurological function of RKIP.

OBJECTIVE:To study the development of changes for Raf kinase inhibitor protein (RKIP) and its mRNA in rats hippocampus after electromagnetic radiation.METHODS:Rats were exposed to X-band high power microwave (X-HPM), S-band high power microwave (S-HPM) and electromagnetic pulse (EMP) radiation source respectively. The animal model of electromagnetic radiation was established. Western blot was used to detect the expression of RKIP, and RT-PCR was applied to detect the expression of RKIP mRNA. The interaction of RKIP and Raf-1 was measured with co-immunoprecipitation method, and the expression of cerebral choline acetyltransferase (CHAT) was measured by immunohistochemistry.RESULTS:The expression of RKIP significantly down-regulated at 6 h after radiation, and recovered at 1 d in group EMP, but the down-regulation continued during 1 approximately 7 d after radiation in the two microwave groups. The expression of RKIP mRNA changed wavily during 6 h approximately 7 d after radiation, which showed down-regulation at 6 h, and up-regulation at 3 d. The interaction of RKIP and Raf-1 decreased during 6 h approximately 7 d after radiation, most significantly at 7 d, and the two microwave groups were more significant. The expression of CHAT decreased continuously during 6 h approximately 7 d after radiation, and generally recovered on 14 d.CONCLUSION:The down-regulation of RKIP and its related proteins of hippocampus is induced by electromagnetic radiation.

Objective To establish the tissue microarray for radiation pulmonary injury in C57BL/6J and C3H/HeN mice and apply this tissue microarray to study the role of FN, LN and a-SMA in the pathogenesis of radiation pulmonary fibrosis. Methods C57BL/6J and C3H/HeN mice were irradiated with 20 Gy 60Co gamma rays on the whole chest. The amount of hydroxyproline in lung was measured. Then the tissue microarray was prepared. The expressions of FN, LN and a-SMA in the lung were detected by immunohistochemistry (IHC) and image analysis. Results The tissue microarray was satisfactorily established, the results of HE and immunohistochemical staining using this tissue microarray were concordant with that of ordinary sections. The changes of the lungs in C57BL/6J mice were as follows: the pathological changes undergone interstitial pneumonia, proliferation and fibrosis. Significant increase of collagen deposition was detected. FN increased significantly on 1 and 3 months after irradiation (p0.01), then decreased gradually to a normal level on 6 months after irradiation. LN elevated gradually after irradiation. The expressions of a-SMA were more intense than that in C3H/HeN mice. The changes of the lungs in C3H/HeN mice were as follows: the pathological changes mainly developed institial pneumonitis, no significant changes of FN expression were observed after irradiation compared to that in the control. LN increased significantly on 1 and 3 months after irradiation, and then decreased gradually. Conclusions The models of pulmonary fibrosis-prone and fibrosis-resistant were established by irradiated with gamma rays. The tissue microarray for radiation pulmonary injury was satisfactorily prepared. The fibrosis-sensitivity of C57BL/6J mice was related with the activation of fibroblasts and the high expression of FN and LN in lung.