Objective To clarify the effects of 5G mobile phone radiofrequency radiation exposure on male mouse fertility and to preliminarily explore the underlying mechanisms. Methods Healthy male C57BL/6 mice aged 7-8 weeks were randomly assigned to Sham group, 3.5 GHz radiofrequency radiation group, and 4.9 GHz radiofrequency radiation group, with 16 mice in each group. The mice were exposed to 3.5 GHz or 4.9 GHz mobile phone radiofrequency radiation for 42 consecutive days (1 h per day). The sperm quality was evaluated using sperm count, deformity rate, and motility. H&E staining was performed to assess testicular tissue structure by observing the morphology of spermatogenic cells at various development stages, the diameter of seminiferous tubules, and the thickness of seminiferous epithelium. The sperm mitochondrial function was assessed using sperm mitochondrial membrane potential and testicular ATP content. The fertility of mice was evaluated through fertility rate, litter size, and survival rate of offspring. The underlying mechanisms were explored by detecting the methylation of LRGUK gene and its mRNA and protein levels. Results Compared with the Sham group, there were no significant changes in sperm count in the 3.5 GHz and 4.9 GHz groups; however, the sperm abnormality rate significantly increased (P < 0.05) and sperm motility significantly decreased (P < 0.05). The structure of testicular tissue, the function of sperm mitochondria, and fertility of mice showed no significant changes as compared with the Sham group. The methylation level of LRGUK gene in the testes significantly increased, while the mRNA and protein expression levels significantly decreased. Conclusion Exposure to 3.5 GHz and 4.9 GHz mobile phone radiofrequency radiation for 42 consecutive days can lead to an increase in sperm deformity rate and a decrease in sperm motility in mice, but has no significant effect on fertility, which may be related to an increase in methylation level of the LRGUK gene in the testes.

Objective:To explore the protective effects of liraglutide on acute lung injury in septic mice and its mechanisms.Methods:Thirty-six male FVB/NJ mice were randomly(random number) divided into three groups: control group (Control, n=12), acute lung injury group (ALI, n=12)and liraglutide intervention group (ALI+LIRA, n=12). Mice model of acute lung injury were prepared by intratracheal instillation of Pseudomonas aeruginosa suspension, while the control group were given intratracheal instillation of equal volume of physiological saline; the mice in ALI+LIRA group were received subcutaneous injection of liraglutide (2 mg/kg) 30 minutes post-induction, while both the mice in control group and ALI group were received subcutaneous injection of equal volume physiological saline. After 24 hours, the mice were euthanized, the lung tissues and bronchoalveolar lavage fluid (BALF) were collected, the lung pathological damage changes were evaluated by hematoxylin eosin staining, the expression of surfactant associated protein D (SP-D)in lung tissue were detected by immunofluorescence assay; total protein concentration in BALF were detected by BCA method, and the levels of interleukin-6 (IL-6) and tumor necrosis factor α(TNF-α)levels in BALF were measured by enzyme-linked immunosorbent assay(ELISA), the protein expression of SP-D in BALF and lung tissue were determined by Western blot. Statistical analysis was performed by SPSS software, and continuous variables were compared with one-way analysis of variance among the groups. Results:Compared with the control group, the mice in ALI group had higher lung histopathology injury score, higher total protein concentration, higher IL-6 and TNF-α levels in BALF, and had less SP-D positive cells in lung tissue; and also had lower expression of SP-D in both BALF and lung tissue, with statistical significance (all P<0.05). Compared with ALI group, the mice in ALI+LIRA group had lower lung histopathology injury score, lower total protein concentration, lower IL-6 and TNF- α levels in BALF, and had more SP-D positive cells in lung tissue; and also had higher expression of SP-D in both BALF and lung tissue, with statistical significance (all P<0.05). Conclusions:Liraglutide attenuates the severity of acute lung injury in septic mice, and its protective mechanism may be associated with the promotion of SP-D secretion.

Objective To study the characteristics of a rat model of migraine with hyperactivity of liver-yang and blood stagnation,which was established by Fuzi Decoction combined with electrical stimulation of the trigeminal ganglion.Methods The 30 SD rats were divided into normal group,model group,and Zhengtian pill group(1.6 g·kg-1),with 10 rats in each group.Rats in the model group and Zhengtian pill group were fed with aconite decoction(2.00 g·kg-1)once a day for 28 day to establish the hyperthermia model;On the 15 day of intragastric administration,rats in the Zhengtai pill group were simultaneously given Zhengtai pill solution once a day for 14 day;At 29 day,the trigeminal ganglion was stimulated to create the migraine model of hyperliver and stasis.After 30 min of the last dose,the macroscopic signs and behaviors were observed,and the blood rheology was detected by blood visticometer.Positive substance P(SP)expression,mRNA and relative protein expression in the trigeminal cervical pulp complex(TCC)in rat mice by immunohistochemistry,RT-qPCR and Western blot analysis.Results Compared with the normal group,the model group has macro signs such as red eyes,irritability,cage fighting,dark tongue and stasis points;regular head positioning and frequent hair management;increased distance in the central area(P<0.05).The relative viscosity of whole blood,plasma viscosity,red blood cell aggregation index all increased(P<0.05),and the red blood cell variant index decreased(P<0.05);SP positive expression,mRNA and the relative protein expression increased in TCC(P<0.05).Compared with model group,Zhengtian pill group can improve macro representation and behavior significantly;can significantly reduce central distance(P<0.05);can reduce plasma viscosity,high-shear relative viscosity of whole blood,and red blood cell aggregation index(P<0.05);increase the Red blood cell variant index significantly(P<0.05);down-regulate SP positive expression,mRNA and relative protein expression in TCC(P<0.05,P<0.01).Conclusion SD rats were used for 4 weeks to construct a migraine model of hyperhepatic hyperactivity and stasis,which was basically consistent with the clinical manifestations.

Sleep is essential for the maintenance of human normal functions.Nowadays,the occurrence of active sleep deprivation(ASD)or passive sleep depriva-tion(PSD)is becoming more and more common due to the inability of the body adapting to the rapid changes in the internal and external environment.SD is not only an action,a brief process or a result,but also a directly or indirectly sustained state,which is associated to sleep time,circadian rhythm or sleep quality.SD can lead to numerous adverse effects on the body,such as sleep-related acute and chronic diseases.Long-term SD increases the risk of neurological and cardiovascular dis-eases as well as immune system dysfunction.In addi-tion,SD may affect the reproductive health of the body,giving rise to a series of potential fertility problems.In recent years,the correlation research and mechanism between SD and the related diseases have become a focus of scholars' attention.Numerous lines of evidence suggest that pathological sleep,such as insomnia and sleep apnea syndrome,is associated with impaired repro-ductive function.Disruptions in the circadian rhythm can also lead to impaired hypothalamic-pituitary-gonadal(HPG)axis function and thereby interfere with the repro-ductive process.Our research team has demonstrated that SD significantly affects the fertility of male and female rats and has adverse effects on reproduction.By new generation sequencing(NGS)and RT-PCR verifica-tion,we have identified differently expressed genes that are involved in mediating the effect of SD on fertility.However,the mechanisms and biological macromolecules regulated by SD are worthy of being further explored.This paper provides a brief review of SD research and then focuses on the adverse impact of SD on fertility,conducting a literature review to sort out the ideas and pro-vide references for research in this field.

Objective:To investigate the effect of radiofrequency radiation (RF) from 5G mobile phone communication frequency bands (3.5 GHz and 4.9 GHz) on the permeability of the blood-brain barrier (BBB) in mice.Methods:A total of 24 healthy adult male C57BL/6 mice (6-8 weeks old) were randomly divided into Sham, 3.5 GHz RF and 4.9 GHz RF groups, and 8 mice in each group. Mice in the RF groups were systemically exposed to 5G cell phone radiation for consecutive 35 d(1 h/d) with 50 W/m 2 power density. The BBB permeability of mice was detected by Evans Blue (EB) fluorescence experiment. The expression levels of the BBB tight junction-related proteins (ZO-1, occludin and claudin-11) and the gap junction-related protein Connexin 43 were determined by Western blot. Results:The number of spots, fluorescence intensity and comprehensive score of EB were significantly increased in 3.5 GHz RF group and 4.9 GHz RF group compared with the Sham group ( t=12.98, 17.82, P<0.001). Compared with the Sham group, the content of S100B in mouse serum was significantly increased in 3.5 GHz RF group and 4.9 GHz RF group ( t=19.34, 14.68, P<0.001). The BBB permeability was increased in the RF group. The expression level of occludin protein was significantly reduced in the 3.5 GHz RF group ( t=-3.13, P<0.05), and this decrease was much profound in the 4.9 GHz RF group ( t=-6.55, P<0.01). But the protein levels of ZO-1, Claudin-11 and Connexin 43 in the cerebral cortex of the RF groups had no significantly difference in comparison with the Sham group( P>0.05). Conclusions:The continuous exposure of mobile phone RF at 3.5 GHz or 4.9 GHz for 35 d (1 h/d) induces an increase of BBB permeability in the mouse cerebral cortex, perhaps by reducing the expression of occludin protein.

OBJECTIVE To expl ore the mechanism of effe ctive fractions from Xiongma decoction in the treatment of migraine with hyperactivity of liver-yang and blood stasis. METHODS Totally 70 male SD rats were randomly divided into normal group ， model group ，positive control group （Flunarizine hydrochloride capsules 0.9 mg/kg），low-dose and high-dose groups of Xiongma decoction effective fractions （ethyl acetate extract 0.87，3.46 g/kg，n-butanol extract 1.80，7.20 g/kg）. Except for normal group ， rats in other groups were given aconite decoction （2 g/kg），once a day ，for 4 consecutive weeks to establish the hyperactivity model of liver-yang. On the 15th day of modeling ，all administration groups were given corresponding drugs intragastrically at the same time ，once a day ，for 2 consecutive weeks. On the 29th day of modeling ，rats trigeminal ganglion was stimulated to establish the migraine model with hyperactivity of liver-yang and blood stasis ，then the medication was maintained for another time according to the above method. The macroscopic signs and behavior of the rats were observed ；positive expression ，mRNA and protein expression of transient receptor potential vanilloid 1 （TRPV1），calcitonin generelated peptide （CGRP），calcitonin receptor-like receptor （CRLR） and receptor-associated membrane proteins 1 （RAMP1） in trigeminal cervical spinal complex （TCC） were detected by immunohistochemistry ，RT-qPCR and Western blot assay. RESULTS Rats in model group showed macrophysical signs and behaviora l manifestations related to migraine with hyperactivity of liver-yang and blood stasis. Thirty minutes after last administration ，the above conditions of rats in Xiongma decoction effective fraction groups were improved significantly. Compared with normal group ， positive expression，mRNA and protein expression of TRPV 1，CGRP， CRLR and RAMP 1 in TCC of rats in the model group were significantly increased （P＜0.05）. Compared with model zengguirong@hnse.org group， most of above indicators in Xiongma decoction effective fraction groups were significantly decreased （P＜0.05）. CONCLUSIONS The mechanism of Xiongma decoction in preventing and treating migraine with hyperactivity of liver-yang and blood stasis may be related to inhibit the activity of TRPV1-CGRP/CGRP receptor signaling pathway in TCC.

Objective:To investigate the effect of thoracic X-ray irradiation on the spermatogenesis of adult male mice.Methods:A total of 24 healthy adult male C57BL/6 mice (6-8 weeks old) were randomly divided into radiation group (Radiation) and sham-radiation group (Sham), 12 mice in each group. The area of thoracic irradiation was 1.5 cm× 2 cm, and the dose rate was 3.04 Gy/min, 8 Gy/d for 3 consecutive days, 24 Gy in total. At 7 d and 21 d after thoracic irradiation, the bilateral testes and epididymal tails were stripped and the testicular index was calculated. The morphology of testis was examined by haematoxylin-eosin (HE) staining, then the diameter of seminiferous tubules and the thickness of seminiferous epithelium were measured. The sperms were collected from the bilateral epididymal tails for sperm counting. The level of apoptosis in testis and levels of apoptosis-related proteins were detected by TUNEL and Western blot, respectively.Results:Compared with Sham group, the morphology of testis and epididymis was seriously damaged, the diameter of seminiferous tubules significantly decreased at 21 d after irradiation ( t = 8.93, P < 0.05), and the seminiferous epithelium significantly decreased at 7 d and 21 d after irradiation ( t = 4.24, 12.77, P < 0.05). In addition, the number of sperms significantly decreased ( t = 4.30, 2.98, P < 0.05). The number of TUNEL positive cells in the seminiferous epithelium significantly increased at 7 d and 21 d after irradiation ( t = -2.73, -3.74, P < 0.05). Meanwhile, the level of cleaved Caspase-3 protein significantly increased at 7 d and 21 d after irradiation ( t = -2.96, -2.46, P < 0.05). The concentrations of SCF and GDNF did not change at 7 d after irradiation, but were significantly increased at 21 d after irradiation ( t = -10.46, -5.42, P < 0.05). Conclusions:The thoracic X-ray irradiation could lead to spermatogenesis disorder in male adult mice, and the induction of spermatogenic cell apoptosis and the secretory dysfunction of sertoli cells may be involved.

Objective: To  investigate  the  effect  of  X-ray  on  the  polarization  of  mouse  microglia  BV-2  cells. Methods: BV-2 cells at the logarithmic growth stage were randomly divided into the Sham irradiation group and 10 Gy irradiation group. The latter group was given a single X-ray irradiation at a dose of 1.28 Gy/min for 7 min 49 s. The activation rate of BV-2 cells was observed and analyzed under a microscope at 1, 3, 6, 24 h and 48 h after irradiation.The changes of cell morphology were observed by HE staining and immunofluorescence staining; The levels of M1-type activation markers (TNF-α and IL-1β) and M2-type activation marker TGF-β1 in the supernatant of BV-2 cells were detected by ELISA. The levels of polarization-related proteins of M1-type (CD86 and iNOS) and M2-type (CD206) in BV-2 cells were detected by Western blotting. Results　: Morphological results showed that BV-2 cells became larger, and their protrude became coarse and shorter, showing "amoeba" like changes after 10 Gy X-ray irradiation. Compared with the Sham group, the activation rate of BV-2 cells was significantly increased at 3 h, and reached the peak at 6 h, and began to recover at 48 h after irradiation. ELISA results showed an obvious increase in the level of TNF-α and TGF-β1 48 h after irradiation.The level of IL-1β showed a transient decrease at 3～6 h, increased at 24 h, and reached the peak 48 h after irradiation. Western blotting results showed that CD86 protein level did not change significantly at each time points after irradiation, and iNOS protein level in- creased significantly at 1, 6, 24 h and 48 h after irradiation. A fluctuating change in CD206 protein level was found after irradiation. Conclusion　 10 Gy X-ray irradiation can induce the activation of BV-2 cells in vitro, and the polarization type changes with the time after irradiation.

Objective:To screen and identify the potential targets of carthamin against sepsis by studying the characteristics of carthamin.Methods:The pharmacological parameters and molecular characteristics of carthamin were analyzed with the aid of Traditional Chinese Medicine Systems Pharmacology (TCMSP). The targets of carthamin were screened by SwissTargetprediction (a website providing compound target prediction) and Drug Repositioning and Adverse drug Reaction via Chemical-Protein Interactome (DRAR-CPI). The anti-sepsis targets were selected from the three databases of Online Mendelian Inheritance in Man (OMIM), Comparative Toxicogenomics Database (CTD) and Therapeutic Targets Database (TTD). The targets of carthamin screened by the two websites and disease targets selected from the three databases were matched to screen the targets of carthamin against sepsis. The anti-sepsis potential targets of carthamin were identified by molecular docking software.Results:The oral bioavailability of carthamin was 41.15%, the drug-likeness was 0.24, and the rotational bond number was 1, which indicated that carthamin was well absorbed by oral administration and showed good drug formation. A total of 115 potential targets of carthamin were screened by SwissTargetprediction and DRAR-CPI; 149 disease targets were found from OMIM, CTD and TTD databases; 115 target proteins of carthamin screened by the two websites were matched with the disease targets , and 10 target proteins were found to be both molecular targets and disease targets. The 10 target proteins were coagulation factor Ⅸ (F9), adenosine A1 receptor (ADORA1), nitric oxide synthase 2 (NOS2), mitogen activity protein kinase 1 (MAPK1), cathepsin G (CTSG), neutrophil elastase (ELANE), protein C (PROC), lipocalin 2 (LCN2), glucose-6-phosphate dehydrogenase (G6PD) and prostaglandin endoperoxidase 2 (PTGS2). Molecular docking software analysis showed that carthamin had the ability to bind to the above 10 target proteins, which were potential targets of carthamin against sepsis. Carthamin could interact with the key amino acid residues of the targeted proteins, so as to play the corresponding efficacy.Conclusion:Carthamin combines with the targets could reduce the tissues and organs damage of sepsis by regulating CTSG, ELANE and LCN2, reduce inflammatory response of sepsis by regulating ADORA1, PTGS2, NOS2, MAPK1 and mediating PROC and F9 to inhibit clotting, and improve oxidative stress, reduce the incidence of sepsis by regulating G6PD, finally, prevented and treated sepsis.

ABSTRACT OBJECTIVE：To study the effects of the extract of Xiongmatang on 5-HT and CGRP in brain tissue of rats with liver-yang hyperactivity and blood stagnation migraine. METHODS：Fifty male spontaneously hypertension （SHR） rats were randomly divided into model control group，Zhengtian pill positive control group （1.6 g/kg），the extract of Xiongmatang low-dose，medium-dose and high-dose groups（4.5，9.0，18.0 g/kg，by crude drug），with 10 rats in each group. Another 10 normal rats were taken as normal control group. Administration group was given relevant medicine intragastrically，once a day，for consecutive 28 d. Normal control group and model control group were given equal volume normal saline intragastrically. After last administration，except for normal control group，other groups were used to stimulate trigeminal ganglia（10 min）to establish liver-yang hyperactivity and blood stagnation migraine model，and maintained the administration once more after making the model. 30 min later，general behavior and tongue quality of rats were observed，and the blood pressure was measured；the contents of 5-HT and CGRP in cerebral tissue of rats were determined by ELISA. The protein expression of CGRP in cerebral tissue of rats were determined by Western blot method. RESULTS： Compared with the normal control group， the rats in the model control group had behavioral symptoms，such as the color of conjunctiva deepened and reddened， excessive hairdressing，head flicking，and most of the rats had purple tongue；the systolic pressure and the content and protein expression of CGRP were all increased obviously（P＜0.05 or P＜0.01）， while the content of 5-HT in cerebral tissue was decreased obviously（P＜0.01）. Compared with model control group，general behavior and tongue quality of rats were improved significantly in administration groups. Systolic pressure，the content and the protein expression of CGRP in cerebral tissue of rats were decreased significantly in Zhengtian pill positive control group and the extract of Xiongmatang high-dose group （P＜0.05），while the content of 5-HT in cerebral tissue of rats were increased significantly in Zhengtian pill positive control group，the extract of Xiongmatang medium-dose and high-dose groups（P＜0.05）.CONCLUSIONS：The extract of Xiongmatang has obvious protective effect on liver-yang hyperactivity and blood stagnation migraine model rats，the mechanism of which may be associated with reducing the content of CGRP in cerebral tissue and raising the content of 5-HT in cerebral tissue