The discontinuation of denosumab[antibody targeting receptor activator of nuclear factor kappa B ligand(RANKL)]therapy may increase the risk of multiple vertebral fractures;however,the underlying pathophysiology is la...The discontinuation of denosumab[antibody targeting receptor activator of nuclear factor kappa B ligand(RANKL)]therapy may increase the risk of multiple vertebral fractures;however,the underlying pathophysiology is largely unknown.In patients who underwent discontinuation after multiple injections of denosumab,the levels of tartrate-resistant acid phosphatase 5b increased compared to pretreatment levels,indicating a phenomenon known as“overshoot.”The rate of decrease in bone mineral density during the withdrawal period was higher than the rate of decrease associated with aging,suggesting that the physiological bone metabolism had broken down.Overshoot and significant bone loss were also observed in mice receiving continuous administration of anti-RANKL antibody after treatment was interrupted,resembling the original pathology.In mice long out of overshoot,bone resorption recovered,but osteoblast numbers and bone formation remained markedly reduced.The bone marrow exhibited a significant reduction in stem cell(SC)antigen 1-and platelet-derived growth factor receptor alpha-expressing osteoblast progenitors(PαS cells)and alkaline phosphatase-positive early osteoblasts.Just before the overshoot phase,the osteoclast precursor cell population expands and RANKL-bearing extracellular vesicles(EVs)became abundant in the serum,leading to robust osteoclastogenesis after cessation of anti-RANKL treatment.Thus,accelerated bone resorption due to the accumulation of RANKLbearing EVs and long-term suppression of bone formation uncoupled from bone resorption leads to the severe bone loss characteristic of denosumab discontinuation.展开更多
Age-related osteoporosis poses a significant challenge in musculoskeletal health;a condition characterized by reduced bone density and increased fracture susceptibility in older individuals necessitates a better under...Age-related osteoporosis poses a significant challenge in musculoskeletal health;a condition characterized by reduced bone density and increased fracture susceptibility in older individuals necessitates a better understanding of underlying molecular and cellular mechanisms.Emerging evidence suggests that osteocytes are the pivotal orchestrators of bone remodeling and represent novel therapeutic targets for age-related bone loss.Our study uses the prematurely aged PolgD257A/D257A(PolgA)mouse model to scrutinize age-and sex-related alterations in musculoskeletal health parameters(frailty,grip strength,gait data),bone and particularly the osteocyte lacuno-canalicular network(LCN).Moreover,a new quantitative in silico image analysis pipeline is used to evaluate the alterations in the osteocyte network with aging.Our findings underscore the pronounced degenerative changes in the musculoskeletal health parameters,bone,and osteocyte LCN in PolgA mice as early as 40 weeks,with more prominent alterations evident in aged males.Our findings suggest that the PolgA mouse model serves as a valuable model for studying the cellular mechanisms underlying age-related bone loss,given the comparable aging signs and age-related degeneration of the bone and the osteocyte network observed in naturally aging mice and elderly humans.展开更多
Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific...Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific animal models induced by inflammatory cytokines.This study introduces a novel mouse model of PD driven by the proinflammatory cytokine CXCL1,identified in our previous research.The involvement of CXCL1 in PD pathogenesis was validated using subacute and chronic MPTP-induced mouse models.Based on these findings,2-month-old C57BL/6J mice were intravenously administered CXCL1(20 ng/kg/day)for 2 weeks(5 days per week),successfully replicating motor deficits and pathological alterations in the substantia nigra observed in the chronic MPTP model.These results demonstrate the potential of CXCL1-induced inflammation as a mechanism for PD modeling.The model revealed activation of the PPAR signaling pathway in CXCL1-mediated neuronal damage by CXCL1.Linoleic acid,a PPAR-γactivator,significantly mitigated MPTPand CXCL1-induced toxicity and reduced serum CXCL1levels.In addition,the CXCL1-injected mouse model shortened the timeline for developing chronic PD mouse model to 2 weeks,offering an efficient platform for studying inflammation-driven processes in PD.The findings provide critical insights into the inflammatory mechanisms underlying PD and identify promising therapeutic targets for intervention.展开更多
Alzheimer'sdisease(AD)isaprogressive neurodegenerative disorder characterized by cognitive impairment and distinct neuropathological features,including amyloid-βplaques,neurofibrillary tangles,and reactive astrog...Alzheimer'sdisease(AD)isaprogressive neurodegenerative disorder characterized by cognitive impairment and distinct neuropathological features,including amyloid-βplaques,neurofibrillary tangles,and reactive astrogliosis.Developing effective diagnostic,preventative,and therapeutic strategies for AD necessitates the establishment of animal models that accurately recapitulate the pathophysiological processes of the disease.Existing transgenic mouse models have significantly contributed to understanding AD pathology but often fail to replicate the complexity of human AD.Additionally,these models are limited in their ability to elucidate the interplay among amyloid-βplaques,neurofibrillary tangles,and reactive astrogliosis due to the absence of spatially and temporally specific genetic manipulation.In this study,we introduce a novel AD mouse model(APP/PS1-TauP301L-Adeno mice)designed to rapidly induce pathological symptoms and enhance understanding of AD mechanisms.Neurofibrillary tangles and severe reactive astrogliosis were induced by injecting AAVDJ-EF1a-hTauP301L-EGFP and Adeno-GFAP-GFP viruses into the hippocampi of 5-month-old APP/PS1 mice.Three months post-injection,these mice exhibited pronounced astrogliosis,substantial amyloid-βplaque accumulation,extensiveneurofibrillarytangles,accelerated neuronal loss,elevated astrocytic GABA levels,and significant spatial memory deficits.Notably,these pathological features were less severe in AAVTauP301L-expressing APP/PS1 mice without augmented reactive astrogliosis.These findings indicate an exacerbating role of severe reactive astrogliosis in amyloid-βplaque and neurofibrillary tangle-associated pathology.The APP/PS1-TauP301L-Adeno mouse model provides a valuable tool for advancing therapeutic research aimed at mitigating the progression of AD.展开更多
Epithelial ovarian cancer(EOC) is the leading cause of gynecological cancer-related mortality in the developed world. EOC is a heterogeneous disease represented by several histological and molecular subtypes. Therefor...Epithelial ovarian cancer(EOC) is the leading cause of gynecological cancer-related mortality in the developed world. EOC is a heterogeneous disease represented by several histological and molecular subtypes. Therefore, exploration of relevant preclinical animal models that consider the heterogenic nature of EOC is of great importance for the development of novel therapeutic strategies that can be translated clinically to combat this devastating disease. In this review, we discuss recent progress in the development of preclinical mouse models for EOC study as well as their advantages and limitations.展开更多
Background:Spinocerebellar ataxia type 2(SCA2)is a neurodegenerative disease marked by significant clinical and genetic heterogeneity,primarily caused by expanded CAG mutations in the ATXN2 gene.The unstable expansion...Background:Spinocerebellar ataxia type 2(SCA2)is a neurodegenerative disease marked by significant clinical and genetic heterogeneity,primarily caused by expanded CAG mutations in the ATXN2 gene.The unstable expansion of CAG repeats disrupts the genetic stability of animal models,which is detrimental to disease research.Methods:In this study,we established a mouse model in which CAG repeats do not undergo microsatellite instability(MSI)across generations.A humanized ATXN2 cDNA with four CAA interruptions within 73 CAG expansions was inserted into the Rosa26 locus of C57BL/6J mice.A 23 CAG control mouse model was also generated to verify ATXN2 integration and expression.Results:In our model,the number of CAG repeats remained stable during transmission,with no CAG repeat expansion observed in 64 parent-to-offspring transmissions.Compared with SCA2-Q23 mice,SCA2-Q73 mice exhibited progressive motor impairment,reduced Purkinje cell count and volume(indicative of cell atrophy),and muscle atrophy.These observations in the mice suggest that the behavioral and neuropathological phenotypes may reflect the features of SCA2 patients.RNA-seq analysis of the gastrocnemius muscle in SCA2-Q73 mice showed significant changes in muscle differentiation and development gene expression at 56 weeks,with no significant differences at 16 weeks compared to SCA2-Q23 mice.The expression level of the Myf6 gene significantly changed in the muscles of aged mice.Conclusion:In summary,the establishment of this model not only provides a stable animal model for studying CAG transmission in SCA2 but also indicates that the lack of long-term neural stimulation leads to muscle atrophy.展开更多
Background:Most mutations in the COL6A3 gene lead to collagen VI-related myopathies.This is due to a reduced expression or mislocalization of the COL6A3 protein.Therefore,studying the consequence of knocking out the C...Background:Most mutations in the COL6A3 gene lead to collagen VI-related myopathies.This is due to a reduced expression or mislocalization of the COL6A3 protein.Therefore,studying the consequence of knocking out the Col6a3 gene in mouse models is relevant,but the Col6a3 mouse models reported so far do not entirely abolish COL6A3 protein expression.Methods:Here,we present the development,validation and preliminary phenotypic characterization of a novel CRISPR-based knockout mouse model targeting Col6a3 exon 3(Col6a3^(d3/d3)).Results:In this mouse model,Col6a3 mRNA is still expressed at a similar level to wild-type littermates,although the expected protein is undetectable by mass spectrometry.Histological analysis of Col6a3^(d3/d3)quadriceps revealed an abnormally high frequency of muscle cells with internally nucleated muscle cells,consistent with a myopathy phenotype.Interestingly,Col6a3^(d3/d3)mice are smaller in size,with their fat,muscle,and bone kept proportional compared to wild-type littermates.Conclusions:In summary,we performed the validation and preliminary phenotypic characterization of a novel Col6a3 knockout mouse model that could be further characterized and used to study COL6A3 biology and model collagen VI-associated diseases.展开更多
Murine subarachnoid hemorrhage(SAH)induced using the filament perforation method is a useful in vivo experimental model to investigate the pathophysiological mechanisms in the brain underlying SAH.However,identifying ...Murine subarachnoid hemorrhage(SAH)induced using the filament perforation method is a useful in vivo experimental model to investigate the pathophysiological mechanisms in the brain underlying SAH.However,identifying mice with comorbid acute neurogenic pulmonary edema(NPE),a life-threatening systemic consequence often induced by SAH,in this model is difficult without histopathological investiga-tions.Herein,we present an imaging procedure involving dual-energy X-ray absorp-tiometry(DXA)to identify NPE in a murine model of SAH.We quantified the lung lean mass(LM)and compared the relationship between micro-computed tomography(CT)evidence of Hounsfield unit(HU)values and histopathological findings of PE.Of the 85 mice with successful induction of SAH by filament perforation,16(19%)had NPE,as verified by postmortem histology.The DXA-LM values correlate well with CT-HU levels(r=0.63,p<0.0001).Regarding the relationship between LM and HU in mice with post-SAH NPE,the LM was positively associated with HU values(r2=0.43;p=0.0056).A receiver operating characteristics curve of LM revealed a sensitivity of 87%and specificity of 57%for detecting PE,with a similar area under the curve as the HU(0.79±0.06 vs.0.84±0.07;p=0.21).These data suggest that confirming acute NPE using DXA-LM is a valuable method for selecting a clinically relevant murine NPE model that could be used in future experimental SAH studies.展开更多
Background:Rabies virus(RABV)-derived neuronal tracing tools are extensively applied in retrograde tracing due to their strict retrograde transsynaptic transfer property and low neurotoxicity.However,the RABV infectio...Background:Rabies virus(RABV)-derived neuronal tracing tools are extensively applied in retrograde tracing due to their strict retrograde transsynaptic transfer property and low neurotoxicity.However,the RABV infection and expression of fluorescence products would be gradually cleared while the infected neurons still survive,a phenomenon known as non-cytolytic immune clearance(NCLIC).This phenomenon introduced the risk of fluorescence loss and led to the omission of a subset of neurons that should be labeled,thereby interfering in the analysis of tracing results.Methods:To compensate for the fluorescence loss problem,in this study,we developed a novel marker footprints(MF)mouse,involving a Cre recombinase-dependent red fluorescent reporter system and systemic expression of glycoprotein(G)and ASLV-A receptor(TVA).Using this mouse model combined with the well-developed RABV-EnvA-ΔG-GFP-Cre viral tool,we developed a novel green-to-red spectral labeling strategy.Results:Neurons in the MF mouse could be co-labeled with green fluorescence from the very quick expression of the viral tool and with red fluorescence from the relatively slow expression of the neuron itself,so neurons undergoing NCLIC with green fluorescence loss could be relabeled red.Furthermore,newly infected neurons could be labeled green and other neurons could be labeled yellow due to the temporal expression difference between the two fluorescent proteins.Conclusions:This is the first polysynaptic retrograde tracing labeling strategy that could label neurons using spectral fluorescence colors with only one injection of the viral tool,enabling its application in recognizing the labeling sequence of neurons in brain regions and enhancing the spatiotemporal resolution of neuronal tracing.展开更多
Craniometaphyseal dysplasia(CMD),a rare craniotubular disorder,occurs in an autosomal dominant(AD)or autosomal recessive(AR)form.CMD is characterized by hyperostosis of craniofacial bones and metaphyseal flaring of lo...Craniometaphyseal dysplasia(CMD),a rare craniotubular disorder,occurs in an autosomal dominant(AD)or autosomal recessive(AR)form.CMD is characterized by hyperostosis of craniofacial bones and metaphyseal flaring of long bones.Many patients with CMD suffer from neurological symptoms.The pathogenesis of CMD is not fully understood.展开更多
Dear Editor,Crimean–Congo hemorrhagic fever(CCHF),caused by the CCHF virus(CCHFV),is a severe tick-borne illness with a wide geographical distribution,posing a significant threat with case fatality rates ranging from...Dear Editor,Crimean–Congo hemorrhagic fever(CCHF),caused by the CCHF virus(CCHFV),is a severe tick-borne illness with a wide geographical distribution,posing a significant threat with case fatality rates ranging from 5%to 70%(Hawman and Feldmann,2023).Due to the lack of approved vaccines and therapeutics,the World Health Organization(WHO)has listed CCHF as one of the priority diseases(Semper et al.,2024).CCHF initially presents as a nonspecific febrile illness,characterized by fever,malaise,myalgia,and nausea,which can rapidly progress to hemorrhagic disease.The hemorrhagic stage is particularly pronounced in severe cases,with rapid progression to disseminated intravascular coagulation(DIC),overt bleeding,kidney or liver failure,and shock(Frank et al.,2024).Up to date,there is an absence of a suitable animal model that can accurately mimic the coagulopathy and bleeding associated with CCHFV infection.Consequently,our understanding of the pathogenic mechanisms underlying these conditions remains limited(Rodriguez et al.,2022).展开更多
Background:It is well recognized that developing new animal models,refining the existing mouse models,and thoroughly characterizing their features are essential for gaining a deeper understanding of rosacea pathogenes...Background:It is well recognized that developing new animal models,refining the existing mouse models,and thoroughly characterizing their features are essential for gaining a deeper understanding of rosacea pathogenesis and for advancing therapeutic strategies in this direction.Accordingly,we aimed to characterize the pathological features of a long-term LL-37-induced mouse model of rosacea and to compare the disease manifestations and pathophysiological characteristics between short-term and long-term LL-37-induced models.A key focus was to investigate differential gene expression and the underlying mechanisms of immune system dysregulation in these models.Methods:We comparatively assessed skin lesion manifestations,the extent of inflammatory infiltration,sebaceous gland alterations,fibrosis,and angiogenesis in both models.Assessments were performed using photographic documentation,hematoxylin-eosin(HE)staining,Van Gieson's(VG)staining,immunohistochemistry,and Western blotting.Furthermore,we employed RNA sequencing to analyze differential gene expression in mouse skin.The RNA sequencing data were validated using immunofluorescence staining and Western blotting,with a specific focus on gene variations and mechanisms related to immune system dysregulation.Results:Mice subjected to long-term LL-37 induction developed rosacea-like pathological features,including angiogenesis,thickened skin tissue,and sebaceous gland hypertrophy.In the short-term LL-37-induced model,immune dysregulation primarily involved the innate immune response.However,long-term LL-37 induction resulted in significant activation of both innate and adaptive immune responses.Conclusion:The long-term LL-37-induced mouse model offers a valuable animal model for the detailed investigation of the pathological mechanisms driving moderate-to-severe rosacea with prolonged disease duration.Importantly,this model provides a significant experimental foundation for exploring the potential role of immune system dysregulation in rosacea pathogenesis.展开更多
Background:New variants of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continue to drive global epidemics and pose significant health risks.The pathogenicity of these variants evolves under immune press...Background:New variants of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continue to drive global epidemics and pose significant health risks.The pathogenicity of these variants evolves under immune pressure and host factors.Understanding these changes is crucial for epidemic control and variant research.Methods:Human angiotensin-converting enzyme 2(hACE2)transgenic mice were in-tranasally challenged with the original strain WH-09 and the variants Delta,Beta,and Omicron BA.1,while BALB/c mice were challenged with Omicron subvariants BA.5,BF.7,and XBB.1.To compare the pathogenicity differences among variants,we con-ducted a comprehensive analysis that included clinical symptom observation,meas-urement of viral loads in the trachea and lungs,evaluation of pulmonary pathology,analysis of immune cell infiltration,and quantification of cytokine levels.Results:In hACE2 mice,the Beta variant caused significant weight loss,severe lung inflammation,increased inflammatory and chemotactic factor secretion,greater mac-rophage and neutrophil infiltration in the lungs,and higher viral loads with prolonged shedding duration.In contrast,BA.1 showed a significant reduction in pathogenicity.The BA.5,BF.7,and XBB.1 variants were less pathogenic than the WH-09,Beta,and Delta variants when infected in BALB/c mice.This was evidenced by reduced weight loss,diminished pulmonary pathology,decreased secretion of inflammatory factors and chemokines,reduced macrophage and neutrophil infiltration,as well as lower viral loads in both the trachea and lungs.Conclusion:In hACE2 mice,the Omicron variant demonstrated the lowest pathogenic-ity,while the Beta variant exhibited the highest.Pathogenicity of the Delta variant was comparable to the original WH-09 strain.Among BALB/c mice,Omicron subvari-ants BA.5,BF.7,and XBB.1 showed no statistically significant differences in virulence.展开更多
Backgroud:Thoracic Trauma and Limb Fractures Are the Two most Common Injuries in Multiple Trauma.However,there Is Still a Lack of Mouse Models of Trauma Combining Tibial Shaft Fracture(TSF)and Thoracic Trauma.In this ...Backgroud:Thoracic Trauma and Limb Fractures Are the Two most Common Injuries in Multiple Trauma.However,there Is Still a Lack of Mouse Models of Trauma Combining Tibial Shaft Fracture(TSF)and Thoracic Trauma.In this Study,we Attempted to Develop a Novel Mouse Model of TSF Combined with Blunt Chest Trauma(BCT).Methods:A total of 84 C57BL/6J male mice were used as the multiple trauma model.BCT was induced by hitting the chests of mice with heavy objects,and TSF was in-duced by hitting the tibia of mice with heavy objects after intramedullary fixation.Serum specimens of mice were received by cardiac puncture at defined time points of 0,6,12,24,48,and 72 h.Results:Body weight and body temperature tended to decrease within 24 h after mul-tiple trauma.Hemoglobin analyses revealed a decrease during the first 24 h after mul-tiple trauma.Some animals died by cardiac puncture immediately after chest trauma.These animals exhibited the most severe pulmonary contusion and hemorrhage.The level of lung damage varied in diverse mice but was apparent in all animals.Classic he-matoxylin and eosin(H&E)-stained paraffin pulmonary sections of mice with multiple trauma displayed hemorrhage and an immunoinflammatory reaction.Bronchoalveolar lavage fluid(BALF)and serum samples of mice with multiple trauma showed an upreg-ulation of interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-1α(TNF-1α)compared with the control group.Microimaging confirmed the presence of a tibia fracture and pulmonary contusion.Conclusions:The novel mouse multiple trauma model established in this study is a common trauma model that shows similar pathological mechanisms and imaging characteristics in patients with multiple injuries.This study is useful for determining whether blockade or intervention of the cytokine response is beneficial for the treat-ment of patients with multiple trauma.Further research is needed in the future.展开更多
Backgroud:Intervertebral disc degeneration(IDD)is one of the common degenerative diseases.Due to ethical constraints,it is difficult to obtain sufficient research on humans,so the use of an animal model of IDD is very...Backgroud:Intervertebral disc degeneration(IDD)is one of the common degenerative diseases.Due to ethical constraints,it is difficult to obtain sufficient research on humans,so the use of an animal model of IDD is very important to clarify the pathogenesis and treatment mechanism of the disease.Methods:In this study,thirty 2-month-old mice were selected for operation to establish a coccygeal IDD model.The distal tail portion of the tail(beyond the 17th coccygeal vertebra)and a small piece of skin above the 8th coccygeal vertebra were excised,and the two incisions were brought together after flexion,and secured with sutures.The heights and signal intensities of the intervertebral discs(IVDs)were assessed using microcomputed tomography(μCT)and magnetic resonance imaging(MRI)at 0,6,12 weeks postoperatively.The overall tissue morphology,cell distribution and density,and extracellular matrix of the IVDs were also assessed using Hematoxylin and Eosin(HE),Safranin O-Fast Green and immunohistochemical staining.Results:All mice in the experimental group survived after the operation,and there were no complications such as wound infection,tail necrosis and suture shedding.The experimental results demonstrated that the suturing method can successfully initiate IDD.Different severity levels of IDD can be induced by controlling the bending angle of the IVDs within the tail loop;however,for consistency,histologic and imaging results should be obtained at the same bending angle and looping period.Conclusions:This IDD model is an effective method for studying the etiology and treatment of degenerative IVD disease.展开更多
Background:Dengue fever,an acute insect-borne infectious disease caused by the dengue virus(DENV),poses a great challenge to global public health.Hepatic involve-ment is the most common complication of severe dengue a...Background:Dengue fever,an acute insect-borne infectious disease caused by the dengue virus(DENV),poses a great challenge to global public health.Hepatic involve-ment is the most common complication of severe dengue and is closely related to the occurrence and development of disease.However,the features of adaptive immune responses associated with liver injury in severe dengue are not clear.Methods:We used single-cell sequencing to examine the liver tissues of mild or se-vere dengue mice model to analyze the changes in immune response of T cells in the liver after dengue virus infection,and the immune interaction between macrophages and T cells.Flow cytometry was used to detect T cells and macrophages in mouse liver and blood to verify the single-cell sequencing results.Results:Our result showed CTLs were significantly activated in the severe liver injury group but the immune function-related signal pathway was down-regulated.The rea-son may be that the excessive immune response in the severe group at the late stage of DENV infection induces the polarization of macrophages into M2 type,and the macrophages then inhibit T cell immunity through the TGF-βsignaling pathway.In ad-dition,the increased proportion of Treg cells suggested that Th17/Treg homeostasis was disrupted in the livers of severe liver injury mice.Conclusions:In this study,single-cell sequencing and flow cytometry revealed the characteristic changes of T cell immune response and the role of macrophages in the liver of severe dengue fever mice.Our study provides a better understanding of the pathogenesis of liver injury in dengue fever patients.展开更多
Background:SARS-CoV-2,first identified in late 2019,has given rise to numerous variants of concern(VOCs),posing a significant threat to human health.The emer-gence of Omicron BA.1.1 towards the end of 2021 led to a pa...Background:SARS-CoV-2,first identified in late 2019,has given rise to numerous variants of concern(VOCs),posing a significant threat to human health.The emer-gence of Omicron BA.1.1 towards the end of 2021 led to a pandemic in early 2022.At present,the lethal mouse model for the study of SARS-CoV-2 needs supplementation,and the alterations in neutrophils and monocytes caused by different strains remain to be elucidated.Methods:Human ACE2 transgenic mice were inoculated with the SARS-CoV-2 proto-type and Omicron BA.1,respectively.The pathogenicity of the two strains was evalu-ated by observing clinical symptoms,viral load and pathology.Complete blood count,immunohistochemistry and flow cytometry were performed to detect the alterations of neutrophils and monocytes caused by the two strains.Results:Our findings revealed that Omicron BA.1 exhibited significantly lower vir-ulence compared to the SARS-CoV-2 prototype in the mouse model.Additionally,we observed a significant increase in the proportion of neutrophils late in infection with the SARS-CoV-2 prototype and Omicron BA.1.We found that the proportion of monocytes increased at first and then decreased.The trends in the changes in the proportions of neutrophils and monocytes induced by the two strains were similar.Conclusion:Our study provides valuable insights into the utility of mouse models for simulating the severe disease of SARS-CoV-2 prototype infection and the milder manifestation associated with Omicron BA.1.SARS-CoV-2 prototype and Omicron BA.1 resulted in similar trends in the changes in neutrophils and monocytes.展开更多
Background:Subcortical ischemic vascular dementia(SIVD)is a common subtype of vascular dementia.Currently,the bilateral common carotid artery stenosis(BCAS)mouse model is the most suitable SIVD rodent model.In this st...Background:Subcortical ischemic vascular dementia(SIVD)is a common subtype of vascular dementia.Currently,the bilateral common carotid artery stenosis(BCAS)mouse model is the most suitable SIVD rodent model.In this study,we investigated the functional and structural impairments in the hippocampus 1 month after BCAS.Methods:We used behavioral tests,laser speckle flowmetry,long-term potentiation,histochemical staining,molecular experiments,and voxel-based morphometry to evaluate the hippocampal impairments.Results:Behavioral studies revealed that BCAS mice exhibited worse performance.Laser speckle flowmetry detected an obvious decrease in cerebral blood flow.The synaptic plasticity of the perforant path-dentate gyrus pathway was inhibited.Decreased fractional anisotropy and increased mean diffusivity were detected in the hippocampus via diffusion tensor imaging data.A reduction in gray matter volume,which was most prominent in the hippocampus and its surrounding areas,was de-tected via voxel-based morphometry analysis.Impairments in cell morphology and myelin integrity were validated using histochemical staining and molecular biology techniques.In addition,the numbers of GFAP+astrocytes and Iba1+microglia in-creased in the hippocampus.Conclusions:Overall,our study demonstrates early functional and structural impair-ments in the hippocampus contributing to learning and memory deficits after 1 month of BCAS,indicating that the hippocampus is vulnerable to chronic cerebral ischemia.展开更多
[Objectives]To explore a new method for induction of allergic rhinitis in mice,and compare and evaluate it with common modeling methods.[Methods]36 mice were randomly divided into the control group,blank group and exp...[Objectives]To explore a new method for induction of allergic rhinitis in mice,and compare and evaluate it with common modeling methods.[Methods]36 mice were randomly divided into the control group,blank group and experimental group,and there were 12 mice in each group.The mice in the control group were conventionally induced.That is,the mice were first injected intraperitoneally with the mixture composed of OVA 50μg,[Al(OH)3]5 mg and 1ml of normal saline once every other day,and then since the 15 th d,20μL of 5%OVA solution was dropped into each nasal cavity once a day,which lasted for 7 d.The blank group was treated with the same amount of normal saline according to the control group,and received intraperitoneal injection and bilateral nasal drip respectively.In the experimental group,mice were first given intraperitoneal injection of the mixture composed of ovalbumin(OVA)75μg,aluminum hydroxide gel[Al(OH)3]8 mg and normal saline 1.5 mL for basic sensitization.On the 26 th d,20μL of 3%OVA solution was dropped into each nasal cavity once a day,which lasted for 10 d.The number of sneezes,the number of nose scratching,the amount of nasal discharge,and the activity of mice in each group were observed,and the behavior of allergic reaction was scored.Meanwhile,the number of eosinophils in the nasal discharge of mice and the IgE content in serum were measured.[Results]The score of nasal stimulation symptoms,the number of eosinophils and serum IgE level of mice in the control group and the experimental group were higher than those in the blank group(P<0.05),and there was no statistical significance between the two groups in the three indicators(P>0.05).[Conclusions]The modeling method was more suitable for the development of allergic rhinitis patients condition,and reduced the probability of death of mice due to modeling,and simplified the experimental operation.展开更多
Cutaneous exposure to food allergens through a disrupted skin barrier is recognized as an important cause of food allergy,and the cutaneous sensitized mouse model has been established to investigate relevant allergic ...Cutaneous exposure to food allergens through a disrupted skin barrier is recognized as an important cause of food allergy,and the cutaneous sensitized mouse model has been established to investigate relevant allergic disorders.However,the role of different genetic backgrounds of mice on immune responses to food allergens upon epicutaneous sensitization is largely unknown.In this study,two strains of mice,i.e.,the BALB/c and C57BL/6 mice,were epicutaneously sensitized with ovalbumin on atopic dermatitis(AD)-like skin lesions,followed by intragastric challenge to induce IgE-mediated food allergy.Allergic outcomes were measured as clinical signs,specific antibodies and cytokines,and immune cell subpopulations,as well as changes in intestinal barrier function and gut microbiota.Results showed that both strains of mice exhibited typical food-allergic symptoms with a Th2-skewed response.The C57BL/6 mice,rather than the BALB/c mice,were fitter for establishing an epicutaneously sensitized model of food allergy since a stronger Th2-biased response and severer disruptions in the intestinal barrier and gut homeostasis were observed.This study provides knowledge for selecting an appropriate mouse model to study food-allergic responses associated with AD-like skin lesions and highlights the role of genetic variations in the immune mechanism underlying pathogenesis of food allergy.展开更多
文摘The discontinuation of denosumab[antibody targeting receptor activator of nuclear factor kappa B ligand(RANKL)]therapy may increase the risk of multiple vertebral fractures;however,the underlying pathophysiology is largely unknown.In patients who underwent discontinuation after multiple injections of denosumab,the levels of tartrate-resistant acid phosphatase 5b increased compared to pretreatment levels,indicating a phenomenon known as“overshoot.”The rate of decrease in bone mineral density during the withdrawal period was higher than the rate of decrease associated with aging,suggesting that the physiological bone metabolism had broken down.Overshoot and significant bone loss were also observed in mice receiving continuous administration of anti-RANKL antibody after treatment was interrupted,resembling the original pathology.In mice long out of overshoot,bone resorption recovered,but osteoblast numbers and bone formation remained markedly reduced.The bone marrow exhibited a significant reduction in stem cell(SC)antigen 1-and platelet-derived growth factor receptor alpha-expressing osteoblast progenitors(PαS cells)and alkaline phosphatase-positive early osteoblasts.Just before the overshoot phase,the osteoclast precursor cell population expands and RANKL-bearing extracellular vesicles(EVs)became abundant in the serum,leading to robust osteoclastogenesis after cessation of anti-RANKL treatment.Thus,accelerated bone resorption due to the accumulation of RANKLbearing EVs and long-term suppression of bone formation uncoupled from bone resorption leads to the severe bone loss characteristic of denosumab discontinuation.
基金the European Research Council(ERC Advanced MechAGE-ERC-2016-ADG-741883)the Swiss National Science Foundation(no.188522).
文摘Age-related osteoporosis poses a significant challenge in musculoskeletal health;a condition characterized by reduced bone density and increased fracture susceptibility in older individuals necessitates a better understanding of underlying molecular and cellular mechanisms.Emerging evidence suggests that osteocytes are the pivotal orchestrators of bone remodeling and represent novel therapeutic targets for age-related bone loss.Our study uses the prematurely aged PolgD257A/D257A(PolgA)mouse model to scrutinize age-and sex-related alterations in musculoskeletal health parameters(frailty,grip strength,gait data),bone and particularly the osteocyte lacuno-canalicular network(LCN).Moreover,a new quantitative in silico image analysis pipeline is used to evaluate the alterations in the osteocyte network with aging.Our findings underscore the pronounced degenerative changes in the musculoskeletal health parameters,bone,and osteocyte LCN in PolgA mice as early as 40 weeks,with more prominent alterations evident in aged males.Our findings suggest that the PolgA mouse model serves as a valuable model for studying the cellular mechanisms underlying age-related bone loss,given the comparable aging signs and age-related degeneration of the bone and the osteocyte network observed in naturally aging mice and elderly humans.
基金supported by the National Natural Science Foundation of China (32471049,32170984,32471188,32200802)Natural Science Foundation of Shandong Province (ZR2023QH110)。
文摘Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific animal models induced by inflammatory cytokines.This study introduces a novel mouse model of PD driven by the proinflammatory cytokine CXCL1,identified in our previous research.The involvement of CXCL1 in PD pathogenesis was validated using subacute and chronic MPTP-induced mouse models.Based on these findings,2-month-old C57BL/6J mice were intravenously administered CXCL1(20 ng/kg/day)for 2 weeks(5 days per week),successfully replicating motor deficits and pathological alterations in the substantia nigra observed in the chronic MPTP model.These results demonstrate the potential of CXCL1-induced inflammation as a mechanism for PD modeling.The model revealed activation of the PPAR signaling pathway in CXCL1-mediated neuronal damage by CXCL1.Linoleic acid,a PPAR-γactivator,significantly mitigated MPTPand CXCL1-induced toxicity and reduced serum CXCL1levels.In addition,the CXCL1-injected mouse model shortened the timeline for developing chronic PD mouse model to 2 weeks,offering an efficient platform for studying inflammation-driven processes in PD.The findings provide critical insights into the inflammatory mechanisms underlying PD and identify promising therapeutic targets for intervention.
基金supported by the National Research Foundation of Korea (NRF)funded by the Ministry of Science,ICT&Future Planning (2022R1A2C2006229,2022R1A6A3A01086868)Korea Dementia Research Project through the Korea Dementia Research Center (KDRC)funded by the Ministry of Health&Welfare and Ministry of Science and ICT,Republic of Korea (RS-2024-00345328)KIST Institutional Grant (2E32851)。
文摘Alzheimer'sdisease(AD)isaprogressive neurodegenerative disorder characterized by cognitive impairment and distinct neuropathological features,including amyloid-βplaques,neurofibrillary tangles,and reactive astrogliosis.Developing effective diagnostic,preventative,and therapeutic strategies for AD necessitates the establishment of animal models that accurately recapitulate the pathophysiological processes of the disease.Existing transgenic mouse models have significantly contributed to understanding AD pathology but often fail to replicate the complexity of human AD.Additionally,these models are limited in their ability to elucidate the interplay among amyloid-βplaques,neurofibrillary tangles,and reactive astrogliosis due to the absence of spatially and temporally specific genetic manipulation.In this study,we introduce a novel AD mouse model(APP/PS1-TauP301L-Adeno mice)designed to rapidly induce pathological symptoms and enhance understanding of AD mechanisms.Neurofibrillary tangles and severe reactive astrogliosis were induced by injecting AAVDJ-EF1a-hTauP301L-EGFP and Adeno-GFAP-GFP viruses into the hippocampi of 5-month-old APP/PS1 mice.Three months post-injection,these mice exhibited pronounced astrogliosis,substantial amyloid-βplaque accumulation,extensiveneurofibrillarytangles,accelerated neuronal loss,elevated astrocytic GABA levels,and significant spatial memory deficits.Notably,these pathological features were less severe in AAVTauP301L-expressing APP/PS1 mice without augmented reactive astrogliosis.These findings indicate an exacerbating role of severe reactive astrogliosis in amyloid-βplaque and neurofibrillary tangle-associated pathology.The APP/PS1-TauP301L-Adeno mouse model provides a valuable tool for advancing therapeutic research aimed at mitigating the progression of AD.
基金supported by the US National Institutes of Health (R01CA160331, R01CA163377, R01CA202919,R01CA239128, P01AG031862, P50CA228991 to R.G.Z. and K99CA241395 to S.K.)US Department of Defense (OC180109 and OC190181 to R.G.Z.)+2 种基金The Honorable Tina Brozman Foundation for Ovarian Cancer Research (to R.G.Z.)Ovarian Cancer Research Alliance Collaborative Research Development Grant (to R.G.Z.)Core facilities support was provided by a Cancer Centre Support Grant(CA010815) to the Wistar Institute。
文摘Epithelial ovarian cancer(EOC) is the leading cause of gynecological cancer-related mortality in the developed world. EOC is a heterogeneous disease represented by several histological and molecular subtypes. Therefore, exploration of relevant preclinical animal models that consider the heterogenic nature of EOC is of great importance for the development of novel therapeutic strategies that can be translated clinically to combat this devastating disease. In this review, we discuss recent progress in the development of preclinical mouse models for EOC study as well as their advantages and limitations.
基金CAMS Innovation Fund for Medical Sciences,Grant/Award Number:CIFMS,2021-I2M-1-024The Joint Fund for the Department of Science and Technology of Yunnan Province-Kunming Medical University,Grant/Award Number:202201AY070001-007+1 种基金Open Research Fund Project of Yunnan Provincial Key Laboratory of Pharmacology of Natural Medicines,Grant/Award Number:YKLPNP-G2403The Science and Technology Leading Talent Program of Yunnan Province,Grant/Award Number:202405AB350002。
文摘Background:Spinocerebellar ataxia type 2(SCA2)is a neurodegenerative disease marked by significant clinical and genetic heterogeneity,primarily caused by expanded CAG mutations in the ATXN2 gene.The unstable expansion of CAG repeats disrupts the genetic stability of animal models,which is detrimental to disease research.Methods:In this study,we established a mouse model in which CAG repeats do not undergo microsatellite instability(MSI)across generations.A humanized ATXN2 cDNA with four CAA interruptions within 73 CAG expansions was inserted into the Rosa26 locus of C57BL/6J mice.A 23 CAG control mouse model was also generated to verify ATXN2 integration and expression.Results:In our model,the number of CAG repeats remained stable during transmission,with no CAG repeat expansion observed in 64 parent-to-offspring transmissions.Compared with SCA2-Q23 mice,SCA2-Q73 mice exhibited progressive motor impairment,reduced Purkinje cell count and volume(indicative of cell atrophy),and muscle atrophy.These observations in the mice suggest that the behavioral and neuropathological phenotypes may reflect the features of SCA2 patients.RNA-seq analysis of the gastrocnemius muscle in SCA2-Q73 mice showed significant changes in muscle differentiation and development gene expression at 56 weeks,with no significant differences at 16 weeks compared to SCA2-Q23 mice.The expression level of the Myf6 gene significantly changed in the muscles of aged mice.Conclusion:In summary,the establishment of this model not only provides a stable animal model for studying CAG transmission in SCA2 but also indicates that the lack of long-term neural stimulation leads to muscle atrophy.
文摘Background:Most mutations in the COL6A3 gene lead to collagen VI-related myopathies.This is due to a reduced expression or mislocalization of the COL6A3 protein.Therefore,studying the consequence of knocking out the Col6a3 gene in mouse models is relevant,but the Col6a3 mouse models reported so far do not entirely abolish COL6A3 protein expression.Methods:Here,we present the development,validation and preliminary phenotypic characterization of a novel CRISPR-based knockout mouse model targeting Col6a3 exon 3(Col6a3^(d3/d3)).Results:In this mouse model,Col6a3 mRNA is still expressed at a similar level to wild-type littermates,although the expected protein is undetectable by mass spectrometry.Histological analysis of Col6a3^(d3/d3)quadriceps revealed an abnormally high frequency of muscle cells with internally nucleated muscle cells,consistent with a myopathy phenotype.Interestingly,Col6a3^(d3/d3)mice are smaller in size,with their fat,muscle,and bone kept proportional compared to wild-type littermates.Conclusions:In summary,we performed the validation and preliminary phenotypic characterization of a novel Col6a3 knockout mouse model that could be further characterized and used to study COL6A3 biology and model collagen VI-associated diseases.
基金supported by the Grants-in-Aid for Scientific Research from Japan Society for the Promotion of Science KAKENHI 22K09110.
文摘Murine subarachnoid hemorrhage(SAH)induced using the filament perforation method is a useful in vivo experimental model to investigate the pathophysiological mechanisms in the brain underlying SAH.However,identifying mice with comorbid acute neurogenic pulmonary edema(NPE),a life-threatening systemic consequence often induced by SAH,in this model is difficult without histopathological investiga-tions.Herein,we present an imaging procedure involving dual-energy X-ray absorp-tiometry(DXA)to identify NPE in a murine model of SAH.We quantified the lung lean mass(LM)and compared the relationship between micro-computed tomography(CT)evidence of Hounsfield unit(HU)values and histopathological findings of PE.Of the 85 mice with successful induction of SAH by filament perforation,16(19%)had NPE,as verified by postmortem histology.The DXA-LM values correlate well with CT-HU levels(r=0.63,p<0.0001).Regarding the relationship between LM and HU in mice with post-SAH NPE,the LM was positively associated with HU values(r2=0.43;p=0.0056).A receiver operating characteristics curve of LM revealed a sensitivity of 87%and specificity of 57%for detecting PE,with a similar area under the curve as the HU(0.79±0.06 vs.0.84±0.07;p=0.21).These data suggest that confirming acute NPE using DXA-LM is a valuable method for selecting a clinically relevant murine NPE model that could be used in future experimental SAH studies.
基金Hubei Natural Science Foundation of China,Grant/Award Number:2024AFB593。
文摘Background:Rabies virus(RABV)-derived neuronal tracing tools are extensively applied in retrograde tracing due to their strict retrograde transsynaptic transfer property and low neurotoxicity.However,the RABV infection and expression of fluorescence products would be gradually cleared while the infected neurons still survive,a phenomenon known as non-cytolytic immune clearance(NCLIC).This phenomenon introduced the risk of fluorescence loss and led to the omission of a subset of neurons that should be labeled,thereby interfering in the analysis of tracing results.Methods:To compensate for the fluorescence loss problem,in this study,we developed a novel marker footprints(MF)mouse,involving a Cre recombinase-dependent red fluorescent reporter system and systemic expression of glycoprotein(G)and ASLV-A receptor(TVA).Using this mouse model combined with the well-developed RABV-EnvA-ΔG-GFP-Cre viral tool,we developed a novel green-to-red spectral labeling strategy.Results:Neurons in the MF mouse could be co-labeled with green fluorescence from the very quick expression of the viral tool and with red fluorescence from the relatively slow expression of the neuron itself,so neurons undergoing NCLIC with green fluorescence loss could be relabeled red.Furthermore,newly infected neurons could be labeled green and other neurons could be labeled yellow due to the temporal expression difference between the two fluorescent proteins.Conclusions:This is the first polysynaptic retrograde tracing labeling strategy that could label neurons using spectral fluorescence colors with only one injection of the viral tool,enabling its application in recognizing the labeling sequence of neurons in brain regions and enhancing the spatiotemporal resolution of neuronal tracing.
基金supported by NIH/NIDCR grant R01DE025664 to IPC.
文摘Craniometaphyseal dysplasia(CMD),a rare craniotubular disorder,occurs in an autosomal dominant(AD)or autosomal recessive(AR)form.CMD is characterized by hyperostosis of craniofacial bones and metaphyseal flaring of long bones.Many patients with CMD suffer from neurological symptoms.The pathogenesis of CMD is not fully understood.
基金supported in part by grants from the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB0490000 to Z.H.)National Key Research and Development Program(2021YFF0702002 to J.L.,2022YFC2303300 to Z.H.,and 2023YFC2305900 to M.W.)+3 种基金“Youth Commando”project(2023QNTJ-02 TO J.L.)Key Project(2024JZZD-02 to Z.H.)of State Key Laboratory of Virology and BiosafetyWuhan Institute of Virology,the National Natural Science Foundation of China(U22A20336 to Z.H.and Y.Z.)Wuhan Natural Science Foundation(202404071010067 to M.W.and 202404071010068 to J.L.).
文摘Dear Editor,Crimean–Congo hemorrhagic fever(CCHF),caused by the CCHF virus(CCHFV),is a severe tick-borne illness with a wide geographical distribution,posing a significant threat with case fatality rates ranging from 5%to 70%(Hawman and Feldmann,2023).Due to the lack of approved vaccines and therapeutics,the World Health Organization(WHO)has listed CCHF as one of the priority diseases(Semper et al.,2024).CCHF initially presents as a nonspecific febrile illness,characterized by fever,malaise,myalgia,and nausea,which can rapidly progress to hemorrhagic disease.The hemorrhagic stage is particularly pronounced in severe cases,with rapid progression to disseminated intravascular coagulation(DIC),overt bleeding,kidney or liver failure,and shock(Frank et al.,2024).Up to date,there is an absence of a suitable animal model that can accurately mimic the coagulopathy and bleeding associated with CCHFV infection.Consequently,our understanding of the pathogenic mechanisms underlying these conditions remains limited(Rodriguez et al.,2022).
基金The National Natural Science Foundation of China,Grant/Award Number:82204006Science and Technology Project of Hebei Education Department,Grant/Award Number:QN2022009+1 种基金Medical Science Research Project of Hebei,Grant/Award Number:20221534National Natural Science Foundation of Hebei Province,Grant/Award Number:H2024209038。
文摘Background:It is well recognized that developing new animal models,refining the existing mouse models,and thoroughly characterizing their features are essential for gaining a deeper understanding of rosacea pathogenesis and for advancing therapeutic strategies in this direction.Accordingly,we aimed to characterize the pathological features of a long-term LL-37-induced mouse model of rosacea and to compare the disease manifestations and pathophysiological characteristics between short-term and long-term LL-37-induced models.A key focus was to investigate differential gene expression and the underlying mechanisms of immune system dysregulation in these models.Methods:We comparatively assessed skin lesion manifestations,the extent of inflammatory infiltration,sebaceous gland alterations,fibrosis,and angiogenesis in both models.Assessments were performed using photographic documentation,hematoxylin-eosin(HE)staining,Van Gieson's(VG)staining,immunohistochemistry,and Western blotting.Furthermore,we employed RNA sequencing to analyze differential gene expression in mouse skin.The RNA sequencing data were validated using immunofluorescence staining and Western blotting,with a specific focus on gene variations and mechanisms related to immune system dysregulation.Results:Mice subjected to long-term LL-37 induction developed rosacea-like pathological features,including angiogenesis,thickened skin tissue,and sebaceous gland hypertrophy.In the short-term LL-37-induced model,immune dysregulation primarily involved the innate immune response.However,long-term LL-37 induction resulted in significant activation of both innate and adaptive immune responses.Conclusion:The long-term LL-37-induced mouse model offers a valuable animal model for the detailed investigation of the pathological mechanisms driving moderate-to-severe rosacea with prolonged disease duration.Importantly,this model provides a significant experimental foundation for exploring the potential role of immune system dysregulation in rosacea pathogenesis.
基金National Science and Technology Infrastructure of China,Grant/Award Number:National Pathogen Resource Center-NPRC-32National Key Research and Development Program of China,Grant/Award Number:2023YFF0724800CAMS Innovation Fund for Medical Sciences,Grant/Award Number:2021-I2M-1-035。
文摘Background:New variants of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continue to drive global epidemics and pose significant health risks.The pathogenicity of these variants evolves under immune pressure and host factors.Understanding these changes is crucial for epidemic control and variant research.Methods:Human angiotensin-converting enzyme 2(hACE2)transgenic mice were in-tranasally challenged with the original strain WH-09 and the variants Delta,Beta,and Omicron BA.1,while BALB/c mice were challenged with Omicron subvariants BA.5,BF.7,and XBB.1.To compare the pathogenicity differences among variants,we con-ducted a comprehensive analysis that included clinical symptom observation,meas-urement of viral loads in the trachea and lungs,evaluation of pulmonary pathology,analysis of immune cell infiltration,and quantification of cytokine levels.Results:In hACE2 mice,the Beta variant caused significant weight loss,severe lung inflammation,increased inflammatory and chemotactic factor secretion,greater mac-rophage and neutrophil infiltration in the lungs,and higher viral loads with prolonged shedding duration.In contrast,BA.1 showed a significant reduction in pathogenicity.The BA.5,BF.7,and XBB.1 variants were less pathogenic than the WH-09,Beta,and Delta variants when infected in BALB/c mice.This was evidenced by reduced weight loss,diminished pulmonary pathology,decreased secretion of inflammatory factors and chemokines,reduced macrophage and neutrophil infiltration,as well as lower viral loads in both the trachea and lungs.Conclusion:In hACE2 mice,the Omicron variant demonstrated the lowest pathogenic-ity,while the Beta variant exhibited the highest.Pathogenicity of the Delta variant was comparable to the original WH-09 strain.Among BALB/c mice,Omicron subvari-ants BA.5,BF.7,and XBB.1 showed no statistically significant differences in virulence.
基金This study was supported by Development Project of Key Laboratory of Big Data Analysis and Knowledge Service of Science and Technology Innovation Platform of Yangzhou-Yangzhou University Cooperation,Grant/Award Number:YBK202204Jiangsu Province 333 High-level Talent.Training Project,Grant/Award Number:BRA2020176。
文摘Backgroud:Thoracic Trauma and Limb Fractures Are the Two most Common Injuries in Multiple Trauma.However,there Is Still a Lack of Mouse Models of Trauma Combining Tibial Shaft Fracture(TSF)and Thoracic Trauma.In this Study,we Attempted to Develop a Novel Mouse Model of TSF Combined with Blunt Chest Trauma(BCT).Methods:A total of 84 C57BL/6J male mice were used as the multiple trauma model.BCT was induced by hitting the chests of mice with heavy objects,and TSF was in-duced by hitting the tibia of mice with heavy objects after intramedullary fixation.Serum specimens of mice were received by cardiac puncture at defined time points of 0,6,12,24,48,and 72 h.Results:Body weight and body temperature tended to decrease within 24 h after mul-tiple trauma.Hemoglobin analyses revealed a decrease during the first 24 h after mul-tiple trauma.Some animals died by cardiac puncture immediately after chest trauma.These animals exhibited the most severe pulmonary contusion and hemorrhage.The level of lung damage varied in diverse mice but was apparent in all animals.Classic he-matoxylin and eosin(H&E)-stained paraffin pulmonary sections of mice with multiple trauma displayed hemorrhage and an immunoinflammatory reaction.Bronchoalveolar lavage fluid(BALF)and serum samples of mice with multiple trauma showed an upreg-ulation of interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-1α(TNF-1α)compared with the control group.Microimaging confirmed the presence of a tibia fracture and pulmonary contusion.Conclusions:The novel mouse multiple trauma model established in this study is a common trauma model that shows similar pathological mechanisms and imaging characteristics in patients with multiple injuries.This study is useful for determining whether blockade or intervention of the cytokine response is beneficial for the treat-ment of patients with multiple trauma.Further research is needed in the future.
基金the Youth Foundation of The 909th Hospital of Xiamen University,Grant/Award Number:22QN001Natural Science Foundation of Fujian Province,China,Grant/Award Number:2019J01144,2022J011482,2023J011839 and 2023J011844+2 种基金National Natural Science Foundation of China,Grant/Award Number:81600696 and 82172477the Fujian Medical University Union Hospital Talent Research Launch Project,Grant/Award Number:2024XH001Natural Science Foundation of Zhangzhou,China,Grant/Award Number:ZZ2024J59。
文摘Backgroud:Intervertebral disc degeneration(IDD)is one of the common degenerative diseases.Due to ethical constraints,it is difficult to obtain sufficient research on humans,so the use of an animal model of IDD is very important to clarify the pathogenesis and treatment mechanism of the disease.Methods:In this study,thirty 2-month-old mice were selected for operation to establish a coccygeal IDD model.The distal tail portion of the tail(beyond the 17th coccygeal vertebra)and a small piece of skin above the 8th coccygeal vertebra were excised,and the two incisions were brought together after flexion,and secured with sutures.The heights and signal intensities of the intervertebral discs(IVDs)were assessed using microcomputed tomography(μCT)and magnetic resonance imaging(MRI)at 0,6,12 weeks postoperatively.The overall tissue morphology,cell distribution and density,and extracellular matrix of the IVDs were also assessed using Hematoxylin and Eosin(HE),Safranin O-Fast Green and immunohistochemical staining.Results:All mice in the experimental group survived after the operation,and there were no complications such as wound infection,tail necrosis and suture shedding.The experimental results demonstrated that the suturing method can successfully initiate IDD.Different severity levels of IDD can be induced by controlling the bending angle of the IVDs within the tail loop;however,for consistency,histologic and imaging results should be obtained at the same bending angle and looping period.Conclusions:This IDD model is an effective method for studying the etiology and treatment of degenerative IVD disease.
基金Chinese Academy of Medical Sciences Initiative for Innovative Medicine,Grant/Award Number:2021-I2M-1-035 and 2022-I2M-1-011。
文摘Background:Dengue fever,an acute insect-borne infectious disease caused by the dengue virus(DENV),poses a great challenge to global public health.Hepatic involve-ment is the most common complication of severe dengue and is closely related to the occurrence and development of disease.However,the features of adaptive immune responses associated with liver injury in severe dengue are not clear.Methods:We used single-cell sequencing to examine the liver tissues of mild or se-vere dengue mice model to analyze the changes in immune response of T cells in the liver after dengue virus infection,and the immune interaction between macrophages and T cells.Flow cytometry was used to detect T cells and macrophages in mouse liver and blood to verify the single-cell sequencing results.Results:Our result showed CTLs were significantly activated in the severe liver injury group but the immune function-related signal pathway was down-regulated.The rea-son may be that the excessive immune response in the severe group at the late stage of DENV infection induces the polarization of macrophages into M2 type,and the macrophages then inhibit T cell immunity through the TGF-βsignaling pathway.In ad-dition,the increased proportion of Treg cells suggested that Th17/Treg homeostasis was disrupted in the livers of severe liver injury mice.Conclusions:In this study,single-cell sequencing and flow cytometry revealed the characteristic changes of T cell immune response and the role of macrophages in the liver of severe dengue fever mice.Our study provides a better understanding of the pathogenesis of liver injury in dengue fever patients.
基金supported by Beijing Natural Science Foundation(Grant No.Z210014)National Natural Science Foundation of China(Grant No.32070543)+1 种基金National Key Research and Development Project of China(Grant No.2022YFC2303404)CAMS Innovation Fund for Medical Sciences(CIFMS)(Grant No.2022-12M-CoV19-002)
文摘Background:SARS-CoV-2,first identified in late 2019,has given rise to numerous variants of concern(VOCs),posing a significant threat to human health.The emer-gence of Omicron BA.1.1 towards the end of 2021 led to a pandemic in early 2022.At present,the lethal mouse model for the study of SARS-CoV-2 needs supplementation,and the alterations in neutrophils and monocytes caused by different strains remain to be elucidated.Methods:Human ACE2 transgenic mice were inoculated with the SARS-CoV-2 proto-type and Omicron BA.1,respectively.The pathogenicity of the two strains was evalu-ated by observing clinical symptoms,viral load and pathology.Complete blood count,immunohistochemistry and flow cytometry were performed to detect the alterations of neutrophils and monocytes caused by the two strains.Results:Our findings revealed that Omicron BA.1 exhibited significantly lower vir-ulence compared to the SARS-CoV-2 prototype in the mouse model.Additionally,we observed a significant increase in the proportion of neutrophils late in infection with the SARS-CoV-2 prototype and Omicron BA.1.We found that the proportion of monocytes increased at first and then decreased.The trends in the changes in the proportions of neutrophils and monocytes induced by the two strains were similar.Conclusion:Our study provides valuable insights into the utility of mouse models for simulating the severe disease of SARS-CoV-2 prototype infection and the milder manifestation associated with Omicron BA.1.SARS-CoV-2 prototype and Omicron BA.1 resulted in similar trends in the changes in neutrophils and monocytes.
基金supported by Tianjin Key Medical Discipline(Specialty)Construction Project(grant number TJYXZDXK-004A).
文摘Background:Subcortical ischemic vascular dementia(SIVD)is a common subtype of vascular dementia.Currently,the bilateral common carotid artery stenosis(BCAS)mouse model is the most suitable SIVD rodent model.In this study,we investigated the functional and structural impairments in the hippocampus 1 month after BCAS.Methods:We used behavioral tests,laser speckle flowmetry,long-term potentiation,histochemical staining,molecular experiments,and voxel-based morphometry to evaluate the hippocampal impairments.Results:Behavioral studies revealed that BCAS mice exhibited worse performance.Laser speckle flowmetry detected an obvious decrease in cerebral blood flow.The synaptic plasticity of the perforant path-dentate gyrus pathway was inhibited.Decreased fractional anisotropy and increased mean diffusivity were detected in the hippocampus via diffusion tensor imaging data.A reduction in gray matter volume,which was most prominent in the hippocampus and its surrounding areas,was de-tected via voxel-based morphometry analysis.Impairments in cell morphology and myelin integrity were validated using histochemical staining and molecular biology techniques.In addition,the numbers of GFAP+astrocytes and Iba1+microglia in-creased in the hippocampus.Conclusions:Overall,our study demonstrates early functional and structural impair-ments in the hippocampus contributing to learning and memory deficits after 1 month of BCAS,indicating that the hippocampus is vulnerable to chronic cerebral ischemia.
基金the National Innovation and Entrepreneurship Project for University Students(202110599012).
文摘[Objectives]To explore a new method for induction of allergic rhinitis in mice,and compare and evaluate it with common modeling methods.[Methods]36 mice were randomly divided into the control group,blank group and experimental group,and there were 12 mice in each group.The mice in the control group were conventionally induced.That is,the mice were first injected intraperitoneally with the mixture composed of OVA 50μg,[Al(OH)3]5 mg and 1ml of normal saline once every other day,and then since the 15 th d,20μL of 5%OVA solution was dropped into each nasal cavity once a day,which lasted for 7 d.The blank group was treated with the same amount of normal saline according to the control group,and received intraperitoneal injection and bilateral nasal drip respectively.In the experimental group,mice were first given intraperitoneal injection of the mixture composed of ovalbumin(OVA)75μg,aluminum hydroxide gel[Al(OH)3]8 mg and normal saline 1.5 mL for basic sensitization.On the 26 th d,20μL of 3%OVA solution was dropped into each nasal cavity once a day,which lasted for 10 d.The number of sneezes,the number of nose scratching,the amount of nasal discharge,and the activity of mice in each group were observed,and the behavior of allergic reaction was scored.Meanwhile,the number of eosinophils in the nasal discharge of mice and the IgE content in serum were measured.[Results]The score of nasal stimulation symptoms,the number of eosinophils and serum IgE level of mice in the control group and the experimental group were higher than those in the blank group(P<0.05),and there was no statistical significance between the two groups in the three indicators(P>0.05).[Conclusions]The modeling method was more suitable for the development of allergic rhinitis patients condition,and reduced the probability of death of mice due to modeling,and simplified the experimental operation.
基金the financial support received from the Natural Science Foundation of China(32202202 and 31871735)the Zhejiang Provincial Natural Science Foundation of China(LGN22C200027)the Open Fund of the Key Laboratory of Biosafety Detection for Zhejiang Market Regulation(2022BS004)。
文摘Cutaneous exposure to food allergens through a disrupted skin barrier is recognized as an important cause of food allergy,and the cutaneous sensitized mouse model has been established to investigate relevant allergic disorders.However,the role of different genetic backgrounds of mice on immune responses to food allergens upon epicutaneous sensitization is largely unknown.In this study,two strains of mice,i.e.,the BALB/c and C57BL/6 mice,were epicutaneously sensitized with ovalbumin on atopic dermatitis(AD)-like skin lesions,followed by intragastric challenge to induce IgE-mediated food allergy.Allergic outcomes were measured as clinical signs,specific antibodies and cytokines,and immune cell subpopulations,as well as changes in intestinal barrier function and gut microbiota.Results showed that both strains of mice exhibited typical food-allergic symptoms with a Th2-skewed response.The C57BL/6 mice,rather than the BALB/c mice,were fitter for establishing an epicutaneously sensitized model of food allergy since a stronger Th2-biased response and severer disruptions in the intestinal barrier and gut homeostasis were observed.This study provides knowledge for selecting an appropriate mouse model to study food-allergic responses associated with AD-like skin lesions and highlights the role of genetic variations in the immune mechanism underlying pathogenesis of food allergy.
