1.Effects of long-term 2.65 GHz radiofrequency radiation on inflammatory response and intestinal microbiota in mice
Keqin LI ; Yanhui HAO ; Ying LIU ; Jun WANG ; Hongyan ZUO ; Hong YANG ; Yang LI ; Hua DENG
Journal of Army Medical University 2025;47(15):1815-1824
Objective To investigate the effects of long-term radiofrequency(RF)radiation at 2.65 GHz on behavior,inflammatory response,and intestinal microecology in mice in order to provide a basis for the safety assessment of long-term RF exposure.Methods One hundred and eight male C57BL/6N mice(17~21 g,6~8 weeks old)were randomly assigned to a control group(Con)and a RF exposure group.The mice of the RF exposure group were subjected to whole-body uniform exposure to 2.65 GHz RF radiation in an electromagnetic reverberation chamber for 3 h/day for 28 consecutive days.RF field distribution and changes in core body temperature were monitored using an electromagnetic radiation analyzer and a fiber-optic temperature probe,respectively.Cognitive function was assessed using the Y-maze and novel object recognition(NOR)test.Anxiety-like behaviors were evaluated through open field test(OFT)and elevated plus maze(EPM),while depressive-like behaviors were examined with sucrose preference test(SPT)and tail suspension test(TST).HE staining was used to observe the histopathological changes in mouse tissues.Radioimmunoassay(RIA)was employed to detect the expression of pro-inflammatory cytokines,TNF-α and IL-1 β,as well as anti-inflammatory cytokines,IL-4 and IL-10 in the serum,brain,jejunum,and spleen samples.Additionally,metagenomic sequencing was performed to assess alterations in the gut microbiota composition.Results Long-term RF radiation led to a maximal increase of 0.59℃in the core body temperature,but had no significant effects on cognitive function,anxiety-like behaviors,or depressive-like behaviors,or apparent damage of the hippocampal or jejunal tissues in the exposed mice.However,RF exposure significantly up-regulated the expression of the pro-inflammatory cytokine TNF-α in serum(P<0.05),and did not significantly alter the concentrations of other cytokines(IL-1β,IL-4,IL-10),caused significant decrease in α-diversity of the intestinal microbiota(P<0.01),with reduced relative abundances of Ligilactobacillus murinus and Acetatifactor muris(P<0.05),while elevated abundances of Lachnospiraceae bacterium(P<0.01).Conclusion Long-term exposure to 2.65 GHz RF radiation induces systemic inflammatory responses and disrupts gut microbiota homeostasis in mice.
2.Roles of oligodendrocytes in cognitive impairment induced by microwave radiation in mice
Jun WANG ; Yujie LIU ; Xuefeng YANG ; Keqin LI ; Ying LIU ; Zhe YANG ; Yumeng YE ; Zehang LIU ; Hongyan ZUO ; Yanhui HAO ; Yang LI
Military Medical Sciences 2025;49(10):738-746
Objective To investigate the functional changes of oligodendrocytes in a mouse model of cognitive impairment induced by microwave radiation and the mechanism.Methods C57BL/6N male mice were exposed to S-band microwave at 2.856 GHz and 8 mW/cm2 for 15 min.The rectal temperature of mice was monitored by an optical fiber thermometer during microwave radiation.The changes of autonomous exploration behavior and learning and memory ability of mice on the 1st and 7th days after microwave radiation were detected via the open field test and novel object recognition test.Immunofluorescence was used to detect the expression and distribution of neuroglia-2 proteoglycan(NG2)and myelin basic protein(MBP)in the hippocampus of mice on the 1st and 7th days after radiation.Clemastine fumarate,a drug that promoted the maturation of oligodendrocyte precursor cells was administered by gavage,and the expression levels of brain-derived neurotrophic factor(BDNF)and fibroblast growth factor 2(FGF2)in hippocampal tissues were detected by radioimmunoassay at 1 and 7 days after radiation.The changes of myelin sheath structure an 1 and 7 days after radiation were observed by transmission electron microscopy.The effects of clemastine fumarate on learning and memory impairment induced by microwave exposure in mice were assessed via open field and new object recognition experiments.Results Under the experimental conditions,the rectal temperature in mice caused by microwave radiation increased by less than 1 ℃,which was within the thermal safety range of the body.The open field test showed that compared with the control group,the microwave radiation group didn't change significant in terms of movement speedon the 1st and 7th days,but the time spent exploring in the central area was significantly reducedon the 1st day after radiation(P<0.05).In the novel object recognition test,the indexes of the mice on the 1st day were significantly reduced(P<0.05),indicating that the anxiety like behavior and cognitive function of the mice were impaired after microwave radiation.Compared with the control group,the proportion of NG2+area in the hippocampus was significantly decreased(P<0.05)in the microwave radiation group,while that of MBP+area hardly changed on the 1st day after microwave radiation(P>0.05).The expression level of oligodendrocyte related BDNF in the hippocampus was significantly decreased(P<0.05).The myelin of the corpus callosum was broken,and the myelin g ratio was significantly increased(P<0.05),suggesting that micro wave radiation could reduce the number of oligodendrocyte precursors and damage the secretion and myelin function of oligodendrocyte.Compared with the radiation group,the expression levels of BNDF and FGF2 in the radiation combined with clemastine fumarate group were up-regulated,the myelin g ratio was significantly decreased on the 1st day after radiation(P<0.05),and the novel object recognition index was significantly increased(P<0.05).Conclusion Pulsed microwave radiation below the body's fever threshold can cause cognitive dysfunction and other brain damage in mice.The impaired secretion and myelin function of oligodendrocytes and the decreased self-repair ability are the important mechanisms of cognitive dysfunction induced by microwave radiation.
3.Intestinal tissue injury and gut microbiome changes in mice treated with whole body irradiation by gamma rays
Min DUAN ; Zhe YANG ; Ying LIU ; Yongyi WANG ; Yumeng YE ; Yanhui HAO ; Hongyan ZUO ; Yang LI
Military Medical Sciences 2024;48(9):641-649
Objective To investigate the effects of 0.5 Gy 60Co γ-ray irradiation on intestinal tissue injury and intestinal microflora in mice.Methods C57BL/6N mice were irradiated with 0.5 Gy 60Co γ-ray at 1 d,3 d,7 d and 14 d after irradiation.Jejunum tissues were fixed and frozen,and feces were frozen.Hematoxylin-eosin staining was used to observe the pathological injury to jejunum after irradiation,ki67 immunohistochemical staining was adopted to detect the proliferation of jejunum crypt cells,and TdT-mediated dUTP nick end labeling was employed to detect the apoptosis of jejunum crypt cells.The expressions of TNF-α,IL-1β,IL-6 and IL-10 cytokines in small intestines were detected via radioimmunoassay.The changes of intestinal flora in mice after irradiation were analyzed by metagenomic sequencing,and LEfSe analysis and ROC analysis were used to screen the bacteria with significant differences.Results After 0.5 Gy 60Co γ-ray irradiation,the proliferative cells of the jejunal crypt were significantly decreased at 1 d after irradiation(P<0.05),while the apoptotic cells were significantly increased at 1 and 3 d after irradiation(P<0.01).The expression of TNF-α at 7 and 14 d after irradiation,that of IL-1 β at 1,3,7 and 14 d after irradiation and that of IL-6 at 3,7 and 14 d after irradiation were significantly increased(P<0.05),while the expression of IL-10 at 7 and 14 d after irradiation was significantly decreased(P<0.05).After 0.5 Gy 60Co γ-ray irradiation,intestinal flora composition changed significantly at phylum,genus and species levels,and Lactobacillus murinus,Lactobacillus johnsonii,Alistipes-unclassified,Mucispirillum schaedleri underwent the most significant changes and had higher LDA scores.Conclusion The whole body irradiation of 0.5 Gy 60Co γ-ray can cause intestinal tissue damage and change the composition of intestinal flora in mice.
4.Effects of 2 650 MHz radiofrequency radiation on the behavior and hippocampal neurotransmitter release of mice
Yujie LIU ; Jun WANG ; Keqin LI ; Chenxu CHANG ; Ying LIU ; Hongyan ZUO ; Yang LI ; Hong YANG ; Yanhui HAO ; Hua DENG
Chinese Journal of Radiological Medicine and Protection 2024;44(5):354-360
Objective:To investigate the effects of 2 650 MHz radiofrequency (RF) exposure on the behavior and neurotransmitter release of mice.Methods:Adult male C57BL/6N mice were divided into a normal control (CON) group and a radiofrequency radiation (RFR) group using the random number table method. The mice in the RFR group were subjected to single-dose whole-body exposure to a uniform 2 650 MHz RF electromagnetic field for 3 h. During the RF exposure, the field strength in the effective working area of the RF radiation platform was measured using an electromagnetic radiation analyzer, and the changes in the anal temperature of the mice were monitored using an optical fiber thermometer. Moreover, the changes in the cognition, social interaction, and emotion of the mice were determined through the new object recognition test, social preference test, and open field test. Finally, the changes in the hippocampal neurotransmitter release levels of the mice were detected using microdialysis sampling and mass spectrometry, and the changes in the hippocampal tissue structure and ultrastructure were observed via microscopy.Results:Under the test conditions, RF radiation improved the anal temperature of the mice, with a maximum increasing amplitude of 0.61℃, falling within the range of thermal safety. The mice in the RFR group experienced a significant decrease in the frequency and time for exploring new objects ( t=4.50, 2.53, P < 0.05) in the new object recognition test, a significant decrease in the frequency ( t=0.08, P<0.01) and time ( t=0.03, P<0.05) for exploring other mice in the social preference test, and no significant change in the frequency and time for exploring the central area ( P > 0.05) in the open field test. Compared to the CON group, the RFR group showed an increase in the release of 5-hydroxytryptamine (5-HT) ( t=-2.56, P < 0.05) and a decrease in the release of acetylcholine (ACh) ( t=2.21, P < 0.05), no significant difference in the release of glutamate (Glu) and γ-aminobutyric acid (GABA) ( P > 0.05), and no evident damage to the hippocampal tissue and structure and synaptic ultrastructure. Conclusions:2 650 MHz RF radiation may induce cognitive impairment and abnormal social preference in mice, which is attributed to neuronal dysfunctions and neurotransmitter release disorders under RF exposure.
5.Histopathological changes in secondary visual cortex and enhanced calcium activity in neurons being involved in microwave radiation-induced anxiety-like behavior
Zhihua FENG ; Ting PAN ; Ganghua HE ; Chenxu CHANG ; Zhilin CUI ; Meiying YANG ; Yanhui HAO ; Fengsong LIU ; Yang LI ; Hongyan ZUO
Chinese Journal of Radiological Medicine and Protection 2024;44(6):464-471
Objective:To clarify the effects of microwave radiation on anxiety-like behavior, the histomorphology of the secondary visual cortex, and calcium activity in neurons.Methods:36 C57BL/6N mice were selected and divided into control group and microwave radiation group according to the random number table method. In the simple behavioral testing, there were 8 mice in the control group and 7 mice in the radiation group. Combining fiber optic recording with behavioral experiments, there were 8 mice in the control group and 7 mice in the radiation group. Hematoxylin-eosin (HE) staining was conducted with 3 mice in each group. A high-power microwave simulated source in the X-band with a center frequency of 9.875 GHz and an average power density of 12 mW/cm 2 was used to irradiate the mice for 15 minutes, establishing a microwave radiation animal model. Then, anxiety-like behavior changes in the radiation group were identified using the open-field and elevated plus maze (EPM) tests. The effects of microwave radiation on the histomorphology of the secondary visual cortex were investigated using HE staining and optical microscopy. Based on the genetically encoded calcium imaging technique, as well as optical fiber recording combined with behavioral paradigms in the open field and the EPM, the changes of calcium activity in neurons in the V2M region of the secondary visual cortex were detected. Results:Compared to the control group, the radiation group showed a significant decrease in the frequency of exploring the central region of the open field and the open arm of the EPM ( t = 2.24, 3.10, P < 0.05). Furthermore, the radiation group exhibited the degeneration and apoptosis of some neurons in the secondary visual cortex, primarily manifested as pyknosis and deep staining, cell body shrinkage, and the slightly widening of perivascular space. Fiber optic recordings and behavioral experiments indicated that compared to the control group, mice in the radiation group exhibited significantly increased calcium activities in neurons of the secondary visual cortex when exploring the central region of the open field ( t = -2.75, P < 0.05) or the open arm of the EPM ( t = -2.77, -3.41, P < 0.05) compared to those before radiation after microwave exposure. Conclusions:Microwave radiation can induce anxiety-like behaviors and histopathological changes in the secondary visual cortex. Increased calcium activity in neurons of the secondary visual cortex is proved to be an important mechanism underlying the changes in anxiety-like behavior due to microwave radiation.
6.Long-term prognostic value of coronary CT angiography-derived fractional flow reserve in elderly patients with CHD
Jingzhou JIANG ; Xiang GUO ; Rui ZUO ; Qian CHEN ; Hongyan QIAO ; Bangjun GUO ; Pengpeng XU ; Tongyuan LIU ; Longjiang ZHANG
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2023;25(12):1255-1259
Objective To evaluate the long-term prognostic value of coronary CT angiography-derived fractional flow reserve(CT-FFR)in elderly patients with coronary heart disease(CHD).Methods A retrospective analysis was performed on 1133 patients with clinically suspected CHD from a prospective observational study based on coronary CTA and CT-FFR at the General Hos-pital of Eastern Theater from April 2018 to March 2019,and 330 elderly CHD patients were even-tually included.According to major adverse cardiovascular events(MACE)occurred or not,295 patients were assigned into non-MACE group and 35 patients into the MACE group.Based on cor-onary CTA data,plaque features were analyzed and CT-FFR values were measured in all lesioned vessels.The relationship of plaque features and CT-FFR with MACE was evaluated by using Cox proportional risk regression model,Kaplan-Meier survival curve,and ROC curve analyses.Results The patients with coronary stenosis(≥50%)or CT-FFR value ≤0.8 had a higher risk of MACE(P<0.01).Univariate Cox analysis showed that coronary stenosis ≥50%and CT-FFR value 0.8 were risk factors of MACE(P<0.01).After adjusting confounding factors,multivariate Cox analysis indicated that CT-FFR ≤0.8(HR=17.037,95%CI:5.060-57.358,P=0.000)was only independent predictor for MACE.The risk prediction model based on CT-FFR presented better performance than the model based on coronary CTA stenosis(C-index:0.820 vs 0.696,P=0.000).Conclusion CT-FFR≤0.8 is an important independent predictor for long-term MACE in elderly CHD patients.Clinical risk stratification based on CT-FFR may optimize prognostic man-agement strategies in these patients.
7.Mechanism of the therapeutic effect of Tetrandrine on striatum injury caused by microwave radiation
Xiaoxu KONG ; Zhihua FENG ; Xuejia WANG ; Ganghua HE ; Ting PAN ; Zhengtao XU ; Yumeng YE ; Yanhui HAO ; Hongyan ZUO ; Yang LI
Chinese Journal of Radiological Medicine and Protection 2023;43(5):328-334
Objective:To study the therapeutic effect Tetrandrine (TET) on striatal injury caused by microwave radiation and underlying mechanism.Methods:C57BL/6N mice were randomly divided into blank control group (C), radiation control group (R), TET group (TET) and TET combined with radiation group (TET+ R). The mice of radiation group were exposed to 2.856 GHz 8 mW/cm2 microwave on whole-body for 15 min. TET (60 mg/kg) was injected intraperitoneally once a day for 3 consecutive days. The TET structure was verified by ultraviolet spectrophotometry. The open field experiment was used to detect the change of anxiety in mice. Histopathological and ultrastructural changes of the striatum were observed by light microscopy and transmission electron microscopy (TMT). Quantitative real-time PCR (qPCR) was used to detect gene expression changes of voltage-gated calcium channel (VGCC) subtype in the striatum.Results:The open field experiments showed that the time and distance of mice to explore the central region after microwave radiation were significantly lower than that before radiation ( t=4.60, 5.18, P<0.01), and the TET administration significantly improved these changes ( F=1.43, 4.37, P < 0.05). 7 d after microwave radiation, some neuronal nuclei in the striatum of mice contracted and could be stained deeply, which was more obvious in the globus pallidus area. The partial neuronal apoptosis, swelling and cavitation of glial cell mitochondria, blurring of synaptic gaps, and widening of perivascular gaps in the striatum were observed by TMT. The above lesions were significantly rescued after TET administration. But both microwave radiation and TET administration had no significant effect on the gene expressions of striatal VGCC ( P > 0.05). Conclusions:TET has a therapeutic effect on anxiety-like behavior and structural damage of striatum caused by microwave radiation, which is independent of the expression of striatal VGCC genes.
8.Influence of microwave radiation on associative learning and memory function and hippocampal structure in mice
Cuicui HU ; Chenxu CHANG ; Xiaoxu KONG ; Yadong LIU ; Yumeng YE ; Yanhui HAO ; Hongyan ZUO ; Yang LI
Chinese Journal of Radiological Medicine and Protection 2022;42(7):481-486
Objective:To investigate the effects of microwave radiation on associative learning and memory function and hippocampal structure of mice.Methods:C57BL/6N mice were ramdomly divided into sham-radiated group ( n=27) and radiation group ( n=2). The radiation group was exposed to microwave at 2.856 GHz, 8 mW/cm 2 for 15 min, then their spatial and associative learning and memory function were examined with the morris water maze and shuttle box behavior experiment. The pathological changes of hippocampal tissue were observed by HE staining and light microscope, the ultrastructural changes of hippocampal tissue were observed by transmission electron microscope. Results:After microwave radiation, the times of mice crossing the platform for the reverse space exploration decreased from(3.60±0.79) times to (2.55±0.47) times( t=2.21, P=0.046), the average active escape rate decreased significantly ( t = 2.70, P<0.05), and the average active latency and the total shock time was significantly prolonged ( t = -3.09, -3.02, P < 0.05). At 8 d after microwave radiation, the nuclei of some neurons in the CA3 and DG regions of the hippocampus were pyknosis. The neurons were apoptotic, the synaptic spaces blurred, the glial cells swollen, and the perivascular spaces widened in the CA3 region of the hippocampus. Conclusions:Microwave radiation can decline the spatial reference memory ability and associative learning and memory ability of mice. The morphological and pathological changes of hippocampus are the structural basis of this dysfunction.
9.Clinical observation of 70 children with severe pneumonia induced by adenovirus type 7
Feiyan CHEN ; Run DANG ; Hongyan PENG ; Wenmin YANG ; Chunmin ZHANG ; Yunlong ZUO ; Jie HONG ; Mingqi ZHAO ; Yi CHEN ; Yiyu YANG
Chinese Pediatric Emergency Medicine 2020;27(8):587-590
Objective:To explore more clinical characteristics and outcomes of children with severe pneumonia induced by adenovirus type 7(Adv-7).Methods:Seventy children with severe pneumonia induced by Adv-7 admitted to the pediatric intensive care unit of Guangzhou Women and Children′s Medical Center from January 2016 to October 2019 were enrolled.The clinical features, treatments and outcomes of these children were recorded.Results:(1)There were 43 males(61.4%)and 27 females(38.6%). Thirty(42.9%)patients were 0-12 months old, 28(40.0%)patients were 13-36 months old, and 12(17.1%)patients were over 36 months old.(2)The average onset time of severe pneumonia induced by Adv-7 was(11.87±7.10)days before being admitted to PICU.The average sequential organ failure assessment score was 6.80±3.13.The average Murray lung injury score was 2.49±1.15.The average P/F value was(150.57±86.25)mmHg(1 mmHg=0.133 kPa). Sixty-four cases(91.4%)were involved in two or more area of lung on X-rays.All cases were diagnosed with sepsis.(3)Laboratory examination: white blood cell count was(7.6±5.5) ×10 9/L, platelet count was(238.8±164.2)×10 9/L, and C-reactive protein was(39.4±37.2)mg/L.(4)Treatments: intravenous gamma globulin was used in 65 cases(92.9%). Intravenous glucocorticoid was used in 45 cases(64.3%). Fiberoptic bronchoscopy was performed in 43 cases(61.4%). Blood purification treatment was performed in 21 cases(30%). Sixty-three patients(90.0%)were treated with non-invasive or invasive ventilator, high frequency ventilator-assisted ventilation was used in 20 cases(28.6%). Surfactant was used in six cases(8.6%). Extracorporeal membrane oxygenation was used in 19 cases(27.1%). (5)The average treatment time for ventilation was(13.10±11.58)days.The average time for temperature recover was(4.69±4.01)days in PICU.The average PICU stay time was(15.76±12.20)days.The average length of stay was(27.04±13.10)days.There were 16 patients died and the mortality was 22.9%. Conclusion:The children with severe pneumonia induced by Adv-7 were critical and had significant lung injuries.Suffering from active clinical treatment such as extracorporeal membrane oxygenation, some of these patients still had poor prognosis.
10.Clinical effect of high-titer plasma in the treatment of severe viral pneumonia in children
Hongyan PENG ; Feiyan CHEN ; Yunlong ZUO ; Run DANG ; Yiyu YANG
Chinese Journal of Emergency Medicine 2020;29(7):970-975
Objective:To explore the clinical effect of high-titer plasma in the treatment of severe viral pneumonia in children.Methods:A total of 80 eligible children with severe viral pneumonia in intensive care unit of Guangzhou Women and Children Medical Center were enrolled in this retrospective study from January 2016 to June 2019, According to whether high-titer plasma was used or not, patients were divided into the high-titer plasma group (40 cases) and non-high-titer plasma group (40 cases). Chi-square test, Fisher's exact probability test and Mann-Whitney U test were used to compare the basic data, infection indicators, blood gas and ventilator parameters related indicators, treatment and prognosis of the two groups. Results:There were no significant differences in age, sex, admission weight, onset days and fever days between the high-titer plasma group and non-high- titer plasma group before entering the study (all P>0.05); There were no significant differences in the worst SOFA score, Murray lung injury score, chest X-ray involvement and sepsis within 3 days (all P>0.05). There were no significant differences in white blood cells, neutrophils and c-reactive protein between the two groups before and after entering the study (day 3 and 7) (all P> 0.05). There were no significant differences in arterial partial pressure of oxygen (PaO 2), partial pressure of carbon dioxide (PaCO 2), lactic acid (Lac), mean airway pressure (MAP), oxygenation index (P/F) and oxygen index (OI) between the two groups before and after entering the study (day 1, 3 and 7) (all P>0.05). There were no significant differences in the use of immunoglobulin, hormone, fiberoptic bronchoscope, blood purification, high frequency ventilator, pulmonary surfactant and extracorporeal membrane between the two groups (all P>0.05). There were no statistical differences in the number of days using invasive ventilator, the number of days staying in PICU and the total length of hospital stay between the two groups (all P>0.05). There was shorter time of the body temperature decreased to normal time in the high-titer plasma group than in the non-high-titer plasma group ( Z=-2.10, P=0.04). The survival rate of the high-titer plasma group was higher than that of the non-high-titer plasma group (85% vs 65%, χ2=4.27, P=0.04). The average daily cost of the high-titer plasma group was less than that of the non-high-titer plasma group, 3 688.38 yuan (2 335.49, 5 741.51) yuan vs 3 979.24 yuan (2 670.68, 9 992.62) yuan, but the difference was not statistically significant (Z=-1.35, P=0.18). No serious adverse reactions were observed during the treatment with high-titer plasma. Conclusions:The treatment of severe viral pneumonia with high-titer plasma can shorten fever time and improve survival rate. High-titer plasma can be used as a safe and effective treatment for severe viral pneumonia.

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