Objective To evaluate the impact of different marine environments on the quality of red blood cells in additive solution with reduced leukocytes transported in a passive blood transport container(2-10℃)in order to provide data for marine transportation of blood.Methods By simulating marine conditions for supply,transportation trials were conducted in both inland and open seas that covered a distance of 20 nautical miles.Paired samplet-tests were used to analyze intra-group changes in quality parameters of red blood cells in additive solution with reduced leukocytes before and after transportation while independent sample t-tests based on these changes(Δ values)were used to study the inter-group differences in severity of damage.Results Intra-group analysis showed that following inland sea transportation,the mean corpuscular volume(MCV)and mean corpuscular hemoglobin(MCH)in these red blood cells were elevated by 0.52%(P<0.05)and 0.68%(P<0.01),with all the indicators within acceptable limits.In contrast,open sea transportation in high seas resulted in a 3.29%increase in the osmotic fragility of erythrocytes(P<0.01),along with a surge of 39.13%in free hemoglobin(FHb)(P<0.01),30.00%in hemolysis rates(P<0.01),and a 0.42%decrease in chloride ion concentrations(P<0.001),with some parameters near critical quality thresholds.Inter-group comparisons confirmed that Δ free hemoglobin(ΔFHb)and Δ hemolysis rates increased more significantly in the open sea group than in the inland sea group(P<0.05),although no statistically significant differences were found in ΔMCV,ΔMCH or Δ chloride ion concentrations(ΔCl⁻)between the two groups.Conclusion This study finds that open-sea transportation causes more significant hemolytic damage to red blood cells in additive solution with reduced leukocytes.The strong mechanical vibrations and impacts resulting from harsh marine conditions may be important determinants of blood quality.It is recommended that anti-shock stabilizing systems be used during long-distance marine transportation of blood and real-time cold-chain monitoring be enforced in the entire transport process to ensure that blood products meet the requirements for clinical quality.
Objective To compare the progression of articular cartilage lesions and subchondral bone microstructural alterations at different postoperative time points among three rat models of osteoarthritis(OA),and to facilitate the selection of OA animal models in research.Methods Sixty-four male Sprague-Dawley rats were randomly assigned to four groups(n=16 per group):a sham group,anterior cruciate ligament transection(ACLT)group,conventional Hulth group,and a group subjected to the conventional Hulth procedure combined with intra-articular sodium iodoacetate injection(Hulth+iodoacetate).Knee joint specimens were harvested at 4 and 8 weeks postoperatively for gross observation and micro-computed tomography(micro-CT).Bone microstructural parameters,including the bone volume fraction(BV/TV),trabecular number(Tb.N),trabecular thickness(Tb.Th),trabecular separation(Tb.Sp),and structure model index(SMI),were quantified via micro-CT analysis.Cartilage structure was observed under light microscopy.Results In each of the model groups,OA phenotypes of varying severity were induced.At 4 weeks postoperatively,roughened articular surfaces,periarticular tissue hyperplasia,and early-stage cartilage damage were detected in the ACLT and Hulth model groups,concurrent with reduced subchondral bone mass and associated osteoporotic changes.At week 8,the disease progressed to the middle and late stages,characterized by severe uneven joint surfaces and the formation of mature osteophytes in the ACLT and Hulth model groups.In the Hulth+iodoacetate group,pathological features of late-stage OA appeared as early as 4 weeks postoperatively.There was extensive cartilage defect with erosion of the subchondral bone.Compared with the sham group,BV/TV and Tb.N were significantly reduced while SMI and Tb.Sp were significantly increased in the Hulth+iodoacetate group(P<0.05).Conclusion Progressive degeneration from early to advanced stages of OA has been recapitulated in the ACLT and Hulth model groups over the 4-8 week period,whereas a severe and highly reproducible OA phenotype can be rapidly induced in the Hulth+iodoacetate model group.Subchondral bone loss,microstructural degeneration,and abnormal bone remodeling have been induced in all model groups.
Objective To characterize the cerebrospinal fluid(CSF)microbiome in patients with central nervous system(CNS)infections and to compare its diversity with healthy controls using metagenomic next-generation sequencing(mNGS).Methods Forty-six patients with CNS infections were enrolled as the infection group while 16 healthy individuals served as the control group.CSF samples were collected and subjected to mNGS.The sequencing data was processed via host sequence depletion,taxonomic profiling,and alpha/beta diversity analyses.Alpha diversity was assessed using the Chao1,Shannon,Simpson,and Pielou indices while beta diversity was explored using principal coordinates analysis(PCoA)combined with permutational multivariate analysis of variance(PERMANOVA).Results A total of 143 microbial species were identified from 46 sequencing samples.Proteobacteria,Actinobacteria,Firmicutes,and Ascomycota were the predominant phyla.Alpha diversity indices were significantly different between the infection group and control group(Chao1,Shannon,and Simpson,all P<0.05),suggesting altered microbial richness and diversity in CNS infections.Beta diversity analysis revealed a clear separation in community structure between the two groups via PCoA and PERMANOVA(P<0.05).Conclusion The diversity and composition of the CSF microbiome differ significantly between those with CNS infections and healthy individuals,suggesting correlations between CNS infections and CSF microbiome features.
Metabolic associated fatty liver disease(MAFLD)can progress to liver fibrosis,leading to an increased risk of related decompensation.Investigation into the mechanisms,early diagnosis,and accurate staging of MAFLD-related liver fibrosis are critical to patient prognosis.MicroRNA,which are involved in post-transcriptional regulation,play significant roles in the initiation and progression of liver fibrosis.They exhibit aberrant expressions across different stages of liver fibrosis and are stably present in body fluids,suggesting their potential as novel circulating biomarkers.This article reviews the research on the mechanisms through which microRNA promote or inhibit MAFLD-related liver fibrosis and on their diagnostic value and therapeutic potential.
Objective To establish a lymphoid organoid model based on human peripheral blood mononuclear cells(PBMCs)and characterize the antigen-specific immune responses induced by the recombinant tetanus neurotoxin heavy chain(TeNT-Hc)protein based on this model.Methods Using viability and apoptosis of in vitro cultured PBMCs as indicators,the effects of key parameters,including different organoid culture methods,extracellular matrix,serum components and seeding densities,on the construction of lymphoid organoid model were compared.Recombinant TeNT-Hc protein was used as the model antigen to stimulate the organoids.Histological staining,flow cytometry and enzyme-linked immunosorbent assay(ELISA)were applied to evaluate the formation of germinal center-like structures,dynamic changes in specific immune cell subsets,and levels of antibody secretion.Results Under the optimized culture conditions,the cell viability of the constructed lymphoid organoids was 65.1%after 16 days of culture.Stimulation of lymphoid-like organoids with the model antigen resulted in marked activation and differentiation of B cell and T cell subsets,accompanied by the expansion of follicular helper T cell subsets.Germinal center-like structures were observed on day 8.Lymphoid organoids mounted B and T cell immune responses upon antigen stimulation,and antigen-specific antibody secretion was detected by day 6.PBMC-derived organoids from five healthy volunteers trended towards immune activation in response to antigen stimulation while retaining inter-individual differences in immune response intensity.Conclusion This established PBMC-derived lymphoid organoid model can effectively induce antigen-specific immune responses and possesses the dual ability to conduct standardized analysis and reflect the immune heterogeneity of the population.It provides a novel research tool for screening of vaccine candidate antigens and for the exploration of humoral immune mechanisms.
Objective To construct interactive biophilic virtual environments using the virtual reality(VR)technology and biophilia theory in order to study their effects on psychophysiological restoration and cognitive enhancement in individuals exposed to confined environments.Methods High-fidelity interactive biophilic virtual environments(forests,oceans,and cities)and a neutral control scenario were created.A randomized controlled trial was conducted in 235 healthy participants.Stress-related physiological responses were assessed using wearable sensors to measure heart rate variability(HRV)and skin conductance levels(SCLs).Cognitive performance was evaluated using the Stroop color-word test and the Alternative Uses Test(AUT).Results Significant differences were observed between the virtual environments.Exposure to the virtual forests resulted in the most pronounced improvements in perceived restoration and positive effect.Compared with the virtual urban and oceanic environments,the forest environment significantly increased HRV/root mean square of successive differences(RMSSD)(P<0.0001)but decreased SCLs(P<0.0001).Furthermore,the Stroop reaction time was significantly reduced(P<0.001),and AUT creativity scores were significantly enhanced(P<0.0001).Conclusion An interactive virtual biophilic forest environment can effectively mitigate the adverse effects of confinement while delivering dual benefits of psychological restoration and cognitive enhancement.This approach has great implications for maintaining long-term operational capability in individuals exposed to prolonged confinement.
The plateau environment poses significant challenges to the physiology and operational capability of military personnel,which is also a key contributor to non-combat casualties.Traditional evaluation methods using single-modal indicators lack dynamic monitoring capabilities and fail to capture complex dynamic stress reactions.This paper reviews the research progress in multimodal data-driven evaluation models for plateau adaptability by analyzing the pathological mechanisms and influencing factors of plateau adaptability and outlining the limitations of current standards and single-modal machine learning.The construction of multimodal data-driven models is explored,focusing on both data architectures that combine physiological,behavioral and environmental modalities for military scenarios and cross-modal alignment and fusion techniques.Furthermore,military applications such as dynamic early warning of non-combat casualties,personalized acclimatization and intelligent military health support are described.Finally,future developments in multimodal data-driven evaluation models for plateau adaptability are predicted in hopes of contributing to the combat effectiveness of troops on the plateau.
Objective To develop positively charged human ferritin nanoparticles capable of modulating the surface properties of adenovirus vectors via electrostatic interaction-mediated self-assembly in order to enhance the cellular delivery efficiency of adenovirus vectors.Methods Based on structure-guided rational design,combinatorial amino acid mutations were introduced into wild-type human ferritin heavy chain(hF-wt)to generate a panel of positively charged ferritin mutants[(+)hFm].The particle size and electrostatic binding capacity of(+)hFm were characterized using dynamic light scattering(DLS)and agarose gel electrophoresis,respectively,to identify the optimal(+)hFm that could self-assemble into nanoparticles and exhibit a high affinity for negatively charged biomacromolecules.Furthermore,the properties of the complex formed by the adenovirus type 5 vector(Ad5)and(+)hFm[Ad5@(+)hFm]were determined.The efficacy with which target transgene expressions were enhanced and the potential cytotoxicity of Ad5@(+)hFm were subsequently evaluated in HeLa cells.Results The positively charged ferritins constructed by combinatorial amino acid mutations successfully self-assembled into nanoparticles with size distributions ranging from 10 to 70 nm.Among these mutants,(+)hFm-2 showed strong electrostatic binding to plasmid DNA at a migration retardation rate of up to 95.59%.At a molar ratio of 1∶1000[Ad5 to(+)hFm-2],the two components formed a complex with modulated surface properties,which significantly improved Ad5-mediated transgene expressions in the cellular model without inducing detectable cytotoxicity.Conclusion The positively charged engineered ferritin variant(+)hFm-2 can efficiently bind to Ad5 via electrostatic interaction,enhance Ad5-mediated transgene expressions,and possess high biocompatibility.
Objective To investigate the efficacy of needle-free injection(NFIT)through cervical lymph nodes as a novel strategy for drug delivery into the brain,and compare its efficiency for different drugs.Methods Using Evans blue as an injectant in vitro,the differences in drug diffusion between NFIT and needle-dependent injection were studied.By using sodium fluorescein as an injectant in vivo,the tissue distribution of sodium fluorescein in mice over time was evaluated after injection by different means before the dynamic changes of drug accumulation in the brain and lymphatic tissues were observed.Three representative drugs-cyanine5.5(Cy5.5,a small molecule compound),Cy5.5-bovine serum albumin(Cy5.5-BSA,a protein drug),and Cy5.5-hyaluronic acid(Cy5.5-HA,a high molecular weight polymer)were used to evaluate the brain delivery efficiency of different drugs via NFIT.Results In vitro diffusion experiments found a more rapid and uniform diffusion of drugs after NFIT while in vivo experiments showed that the delivery efficiency of fluorescein sodium through intracervical lymphatic NFIT was significantly higher in the cortex and hippocampus of mice than with traditional injection methods.A comparison of three representative drugs confirmed that intracervical lymphatic NFIT was an effective way to deliver small molecule compounds and protein drugs to the brain,but was not workable for polymers of high molecular mass.Conclusion Intracervical lymphatic NFIT can efficiently mediate the brain delivery of small molecules and protein drugs,which promises to be a drug delivery method for the application of such drugs to the brain.
Flight fatigue can impact the ability of pilots to complete long endurance cross-domain flight missions,and skeletal muscle fatigue caused by insufficient muscle endurance is an important cause of flight fatigue.Circadian rhythm and clock genes reportedly play an important role in regulating muscle endurance,and BMAL1 is one of the core clock genes that affect muscle endurance by regulating muscle metabolism,energy production,and muscle fiber-type switching.Disturbance of circadian rhythm or abnormal expressions of BMAL1 can lead to decreased muscle endurance,resulting in body fatigue.In this paper,the research progress in the role of clock gene BMAL1 in regulating muscle endurance is reviewed,which is expected to provide insights and methods for improving muscle endurance and anti-fatigue ability of pilots in long-endurance cross-domain flights.
Current methods for bioterrorism detection and monitoring have limitations,highlighting the urgent need for a rapid,on-site and real-time detection tool as a complementary approach.Biomedical detection dogs can serve as an effective means of detecting and monitoring bioterrorism attacks,thereby providing technical support for early warning and prevention.This article reviews the olfactory mechanisms of canines,analyzes the types of bioterrorism agents and characteristics of their odor signatures,and explores the applications,advantages,and potential scenarios of using biomedical detection dogs in countering bioterrorism.Strategic recommendations are also offered in hopes of providing a reference for the application of sniffer dogs in bioterrorism detection.
目的 调查中国维和医疗分队空运后送对象分布特点,为促进维和空运医疗后送行动时效与安全提供参考.方法 回顾性调查2003-2018年刚果(金)、苏丹、马里、黎巴嫩等4个联合国维和任务区空运医疗后送情况,描述性统计分析空运伤病员的伤情分布与疾病种类特点.结果 在811例空运伤病员中,疾病占比66.3%,创伤占比33.7%.创伤病例中,脑损伤、骨折合计占比超50%.疾病病例中,非传染性疾病占比64.5%,传染性疾病占比35.5%.结论 维和空运后送对象具有多样化特点,既有常见脑损伤、骨折等创伤伤员,又有疟疾、肺结核、艾滋病等传染病病员.建议加强颅脑与骨创伤医护力量及卫生防疫,针对后送对象差异灵活调整机载药材物资编配,严格落实空运医疗后送业务操作规范,提升多元文化与冲突环境伤病员空运的时效与安全.
Objective To investigate the dynamic changes in cardiac structure and function at different time points within one week following high-power microwave exposure in rats in order to provide data for formulating protective strategies and elucidating the mechanisms of microwave-induced cardiac injury.Methods Male Wistar rats were exposed to high-power microwave radiation in the S-band(2.856 GHz)at an average power density of 50 mW/cm²for 30 min.At 1,3,and 7 days post-radiation,cardiac electrophysiological function,macroscopic morphology,and microscopic structure were assessed using serum biochemical analysis(the cardiac enzyme profile,injury markers,and ion concentrations),electrocardiography(ECG),echocardiography,hematoxylin-eosin(HE)staining,and transmission electron microscopy.Results Rectal temperatures significantly increased in rats after microwave irradiation.Biochemical results of the serum showed significant increases in levels of aspartate aminotransferase,lactate dehydrogenase,and creatine kinase at 1 and 7 days post-microwave exposure.Peak changes in fatty acid-binding protein,cardiac troponin T,N-terminal pro-B-type natriuretic peptide,and serum calcium and potassium ion concentrations occurred at 3 days post-exposure,indicating maximal injury at this time point.ECG revealed decreased heart rate,prolonged R-R intervals,and shortened corrected QT intervals at 1 day post-irradiation.Three days after exposure,heart rate increased and P-wave amplitude was elevated,but all parameters largely returned to control levels by day 7.Echocardiography suggested significant thickening of the interventricular septum and left ventricular wall,along with increased left ventricular mass starting at 3 days post-irradiation.These structural alterations persisted until day 7.Histopathological and ultrastructural observations confirmed that myocardial injury peaked at 3 days post-radiation,characterized by inflammatory cell infiltration,myofibrillar disarray,mitochondrial swelling and vacuolation,and disruption of sarcomere structure.These lesions were mitigated by day 7,but complete recovery did not occur.Conclusion 2.856 GHz high-power microwave radiation induces acute cardiac injury in rats,manifested as myocardial enzyme leakage,ion homeostasis imbalance,abnormal electrical activity,and tissue structural remodeling.The effect of injury occurs in a time-dependent manner and peaks at 3 days post-radiation.By day 7,there is partial recovery,yet residual structural abnormalities persist.
Objective To investigate the transcription of neutrophils(Neu)expressing high levels of Csf3rhiCD14hiNeu in C3H/HeN mouse lung tissues before and after irradiation.Methods Twelve C3H/HeN mice were randomly divided into an irradiation group and a control group before being exposed to a single dose of 20 Gy chest irradiation.Twenty-four weeks after irradiation,two Neu subsets,Csf3rhiCD14hiNeu and Csf3rlowCD14hiNeu,were selected from lung tissues of irradiated and non-irradiated mice by flow cytometry.Transcriptomic sequencing(RNA-Seq)was performed on the two types of cells.Differentially expressed genes(DEGs)were identified using the DESeq2 package.Functional enrichment analysis and gene set enrichment analysis(GSEA)were performed using the gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)databases.Results At 24 weeks post-irradiation,the alveolar space was significantly reduced,collagen deposition increased,and fibrosis localized in C3H/HeN mice.RNA-seq analysis suggested that there were 1499 DEGs in Csf3rhiCD14hi Neu compared with Csf3rlowCD14hi Neu in the irradiated group,179 of which were upregulated and 1320 downregulated.In the control group,there were 701 significantly upregulated and 431 downregulated genes in Csf3rhiCD14hi Neu compared with Csf3rlowCD14hi Neu.Compared with non-irradiated Csf3rlowCD14hi Neu,there were 1319 upregulated and 219 downregulated genes in the irradiated group.Compared with non-irradiated Csf3rhiCD14hi Neu,the irradiated group had 202 upregulated and 540 downregulated genes.GO enrichment analysis indicated that these DEGs were involved in immune response,Wnt signaling pathway,and cytokine production.KEGG pathway analysis pointed to significant enrichment in the TGF-β signaling pathway,cytokine-cytokine receptor interactions,and Jak-STAT signaling pathway.GSEA revealed significant alterations in non-canonical NF-κB signaling,PD-1 signaling,and laminin interaction-related pathways.Conclusion Thoracic γ-ray irradiation can cause pulmonary fibrosis in C3H/HeN mice,with marked transcriptomic differences between Csf3rhiCD14ʰⁱand Csf3rlowCD14hi Neu subsets in lung tissue post-irradiation.
Objective To explore the effect of knockdown of keratinocyte associated protein 2(KRTCAP2)on ricin-induced apoptosis and the related mechanism.Methods Small interfering RNA(siRNA)transfection was used to establish siNC and siKRTCAP2 cell lines in the cervical cancer cell line HeLa.Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to detect the mRNA expression level of KRTCAP2.Western blot assay was used to evaluate the levels of autophagy and apoptosis.The monodansylcadaverine(MDC)staining kit for cellular autophagy was applied to detect the formation of autophagosomes in siRNA-transfected cells.Cell Counting Kit-8(CCK-8)assay and colony formation assay were conducted to determine cell viability.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining was used to assess cell apoptosis.Results Knockdown of KRTCAP2 significantly enhanced autophagy.Compared with the siNC group,the mRNA expression level of KRTCAP2 was markedly decreased in the siKRTCAP2 group,in which autophagy was significantly elevated,along with significant upregulation of fluorescent signals of autophagosomes.Ricin induced the death of wild-type HeLa cells in a concentration-dependent manner and inhibited cellular autophagy.Knockdown of KRTCAP2 improved cellular resistance to ricin by enhancing autophagy.Compared with the siNC group,autophagy kept increasing while cell viability was enhanced under ricin induction in the siKRTCAP2 group.Chloroquine(CQ),an autophagy inhibitor,partially blocked the resistance of siKRTCAP2-transfected cellsto ricin,accompanied by reduced autophagy and cell viability.In addition,knockdown of KRTCAP2 mitigated ricin-induced cell apoptosis.Conclusion Knockdown of KRTCAP2 can enhance autophagy while ameliorating cell apoptosis induced by ricin,thereby exerting a protective effect on cells.
Objective To design and synthesize a mannose-modified PLGA-PEG nanodrug delivery system capable of brain-targeting accumulation and blood-brain barrier(BBB)permeation.Methods bEnd.3 mouse vascular endothelial cells were used to establish a monolayer cell model using Transwell chambers.The barrier integrity was evaluated by measuring the transendothelial electrical resistance before an in vitro BBB model was established.A post-traumatic stress disorder model was established to find out about the in vivo BBB permeability of nanomaterials.In the in vitro experiment,nanomaterials were delivered to BV2 cells while in the in vivo experiment,nanomaterials were injected into the tail vein of C57BL/6 mice.The ability of nanomaterials to penetrate the blood-brain barrier was detected by a spectrophotometer,laser confocal microscopy and flow cytometry.The distribution and rate of brain enrichment of nanomaterials were dynamically monitored by a small animal in vivo near-infrared fluorescence imaging system.Results In vitro experiments showed that the PPM system loaded with a fluorescent molecule,purpurin 18(P18),could effectively penetrate the BBB with a fluorescence permeability of 7%.The system was capable of both prolonged retention in the brain with a brain accumulation rate of 30%,and significant brain enrichment in a post-traumatic stress disorder(PTSD)mouse model with an enrichment efficiency of 54%.Conclusion The newly developed BBB-penetrating nano-delivery system can ensure effective BBB penetration and targeted delivery in PTSD mouse models,with significant implications and translational potential for neuropharmaceutical development.
The high incidence of exertional heat stroke(EHS)has posed a huge threat to the health and combat capability of military personnel.This paper reviews recent advancements in the prevention and management of EHS in general and three pivotal interventions—heat acclimatization,hydration strategies,and cooling interventions—in particular.An optimized framework for military medical support to EHS is recommended.In terms of heat acclimatization,it is recommended that China's military learn from short-term training protocols employed inother countries,and that a hybrid model combining basic acclimatization with short-term intensive training be adopted.Moreover,intelligent monitoring technologies are to beused as an alternative to traditional laboratory-based heat tolerance tests.Hydration strategies call for individualized regimensand the adherence to the principle of"high-frequency and small-volumes"of fluid intake.Cooling interventions ought to revolve around the critical"golden 30 minutes"therapeutic window,with cold water immersion(CWI)designated as the first option.Additionally,research that aims to investigate the viability of seawater as a CWI alternative is proposed.Based on foreign experience and the realities in China,this paper recommends a comprehensive,integrated and tripartite strategy that involves acclimatization,hydration,and cooling for the whole-process prevention and control of EHS.This approach is expected to provide data for mitigating EHS during military operations in hot and humid environments and for enhancing the efficiency of military medical support.
Gut microbiota(GM)-derived metabolites play a crucial regulatory role in the occurrence and development of colon cancer.These metabolites,including short-chain fatty acids,bile acid metabolites,nitrogen-containing compounds,and genotoxic substances,can influence colon cancer progression through multiple mechanisms,such as modulation of intestinal barrier function,DNA damage and repair,inflammatory responses,and immune regulation.Dietary patterns can make a difference in colon cancer risk by altering gut microbiota composition and metabolic activity.In recent years,diagnostic and therapeutic strategies based on GM-derived metabolites,including targeted metabolite interventions,personalized nutritional interventions,and microecological regulation,have promised broad clinical applications.This review summarizes the classification and sources of GM-derived metabolites,explores their mechanisms of action in colon cancer,and focuses on the role of the"diet-gut microbiota-metabolite"axis in colon cancer development.In addition,it highlights the clinical translational potential of GM-derived metabolites and current challenges.
Objective To investigate the real-time biological effects of 2650 MHz radiofrequency radiation(RFR)on learning and memory as well as calcium activity patterns of hippocampal CA1 pyramidal neurons of mice.Methods Adult male C57BL/6N mice were randomly assigned to control(CON)and RFR groups before being exposed to 2650 MHz RFR for 3 hours in an electromagnetic reverberation chamber.Within 1 hour after exposure,changes in learning and memory were assessed using novel object recognition(NOR),object location recognition(OLR),and temporal order recognition(TOR)tests.Calcium activity of CA1 pyramidal neurons was monitored before,during,and after exposure using genetically encoded calcium imaging combined with fiber photometry.Neuronal activation was evaluated by c-Fos immunofluorescence staining.Results Compared with the CON group,RFR-exposed mice showed significantly reduced preference indices for target objects in the NOR(P<0.001),OLR(P<0.01),and TOR(P<0.01)tests within 1 hour post-exposure.During RFR exposure,neuronal calcium signals exhibited an abnormal high-frequency but low-amplitude pattern,characterized by increased event frequency(P<0.05)and decreased single-event amplitude(P<0.05).Furthermore,the immunofluorescence intensity of c-Fos in the hippocampal CA1 region was significantly reduced(P<0.05).Conclusion 2650 MHz radiofrequency radiation exposure can induce immediate effects,such as abnormalities in calcium signaling patterns and suppressed excitability of pyramidal neurons in the hippocampal CA1 region,ultimately leading to impaired learning and memory functions.
Objective To establish and validate a sensitive and reliable ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for quantitative analysis of cepharanthine in rat plasma.Methods Plasma samples were treated using the protein precipitation method.Chromatographic separation was performed on column maintained at 50℃.Gradient elution was employed with mobile phases composed of(A)0.05%formic acid aqueous solution containing 1 mmol/L ammonium formate and(B)0.1%formic acid in acetonitrile at a flow rate of 0.40 mL/min.Mass spectrometric detection was carried out using an electrospray ionization(ESI)source in a positive-ion mode with multiple reaction monitoring(MRM).Mass transition m/z 607.3→576.2 was adopted for cepharanthine and m/z 260.2→116.1 for propranolol,the internal standard.This method was validated and used to quantify cepharanthine in plasma samples collected from rats following a single intragastric administration to investigate its pharmacokinetic profile.Results Cepharanthine proved to be linear in rat plasma over the concentration range of 0.1 to 80 ng/mL.The intra-and inter-batch accuracies ranged from-0.43%to 11.73%,and precisions from 3.04%to 13.83%.The recovery rates of quality control samples ranged from 86.70%to 88.48%.The accuracy and precision met the standards across six lots of the matrix.The quality control samples remained stable under the following conditions:24 hours of storage in the autosampler at 15℃,followed by three freeze-thaw cycles,and 10 days of storage at-20℃.All these met the requirements for bioanalytical analysis of samples.Pharmacokinetic studies showed that the time to maximum concentration(Tmax)was 6 to 12 h,the maximum concentration(Cmax)was(12.31±3.78)ng/mL,the terminal elimination half-life(t1/2)was(10.07±0.66)h,and the mean residence time(MRT0-t)was(24.80±1.98)h after intragastric administration of 3 mg/kg cepharanthine to rats.Conclusion This study has established a UPLC-MS/MS method for quantitative analysis of cepharanthine in the rat plasma matrix,which is simple and sensitive,with a lower limit of quantification of 0.1 ng/mL.
