Objective:To investigate the sterilizing potential of zinc gluconate,calcium chloride(CaCl_(2))and cadmium chloride(CdCl_(2))following a single intratesticular administration in adult male rats.Methods:60 adult male W...Objective:To investigate the sterilizing potential of zinc gluconate,calcium chloride(CaCl_(2))and cadmium chloride(CdCl_(2))following a single intratesticular administration in adult male rats.Methods:60 adult male Wistar albino rats(Rattus norvegicus)weighing 160-200 g and aged 5-6 months randomly received a single intratesticular injection of normal saline(Group A),zinc gluconate 13.3 mg/mL plus L-Arginine(Group B),20%CaCl_(2)(Group C),and CdCl_(2)0.5 mg/kg body weight plus ethylenediaminetetraacetic acid(EDTA)(Group D),respectively,along the entire route from the caudoventral aspect of each testis.They were euthanized up to 180 days to evaluate reproductive tract toxicology.Results:The reproductive organ weights were markedly reduced,with testes severely atrophied in group B,pea-sized and stony hard in group C,and moderately reduced in group D.Azoospermia was evident in groups B and C,while sperm concentration was reduced to<1 million/mL with zero sperm motility in group D.Rats of groups B and C failed to show mounting and copulatory behaviour.A completely disorganized mesh of cellular elements was observed in the seminiferous tubules of group B,while pyknotic germ cell and arrest of spermatogenesis,exfoliated germ cells,occasional syncytial bodies and smaller Leydig cells were evident in groups C and D.Significantly reduced testosterone levels,increased luteinizing hormone(LH)and follicle-stimulating hormone(FSH)levels,returned to normal after 90 days in group D.Conclusions:Zinc gluconate and CaCl_(2)injections severely affected the reproductive organs and libido and rats treated with CdCl_(2)exhibited diminished spermatogenesis with normal libido.Thus,the need-based selection of intratesticular agents should consider their distinct effects on spermatogenesis,libido,and hormonal balance for achieving targeted sterilization outcomes.展开更多
Our previous study demonstrated that combined transplantation of bone marrow mesenchymal stem cells and retinal progenitor cells in rats has therapeutic effects on retinal degeneration that are superior to transplanta...Our previous study demonstrated that combined transplantation of bone marrow mesenchymal stem cells and retinal progenitor cells in rats has therapeutic effects on retinal degeneration that are superior to transplantation of retinal progenitor cells alone.Bone marrow mesenchymal stem cells regulate and interact with various cells in the retinal microenvironment by secreting neurotrophic factors and extracellular vesicles.Small extracellular vesicles derived from bone marrow mesenchymal stem cells,which offer low immunogenicity,minimal tumorigenic risk,and ease of transportation,have been utilized in the treatment of various neurological diseases.These vesicles exhibit various activities,including anti-inflammatory actions,promotion of tissue repair,and immune regulation.Therefore,novel strategies using human retinal progenitor cells combined with bone marrow mesenchymal stem cell-derived small extracellular vesicles may represent an innovation in stem cell therapy for retinal degeneration.In this study,we developed such an approach utilizing retinal progenitor cells combined with bone marrow mesenchymal stem cell-derived small extracellular vesicles to treat retinal degeneration in Royal College of Surgeons rats,a genetic model of retinal degeneration.Our findings revealed that the combination of bone marrow mesenchymal stem cell-derived small extracellular vesicles and retinal progenitor cells significantly improved visual function in these rats.The addition of bone marrow mesenchymal stem cell-derived small extracellular vesicles as adjuvants to stem cell transplantation with retinal progenitor cells enhanced the survival,migration,and differentiation of the exogenous retinal progenitor cells.Concurrently,these small extracellular vesicles inhibited the activation of regional microglia,promoted the migration of transplanted retinal progenitor cells to the inner nuclear layer of the retina,and facilitated their differentiation into photoreceptors and bipolar cells.These findings suggest that bone marrow mesenchymal stem cell-derived small extracellular vesicles potentiate the therapeutic efficacy of retinal progenitor cells in retinal degeneration by promoting their survival and differentiation.展开更多
AIM： TO characterize a culture model of rat CCA cells, which were derived from a transplantable TTA-induced CCA and designated as Chang Gung CCA （CGCCA）. METHODS： The CGCCA cells were cultured at in vitro passage ...AIM： TO characterize a culture model of rat CCA cells, which were derived from a transplantable TTA-induced CCA and designated as Chang Gung CCA （CGCCA）. METHODS： The CGCCA cells were cultured at in vitro passage 12 times on a culture dish in DMEM medium. To measure the doubling time, 103 cells were plated in a 96-well plate containing the growth medium. The cells were harvested 4 to 10 d after seeding, and a standard MTT assay was used to measure the growth. The phenotype of CACCA cell and xenograft was determined by immunohistochemical study. We also determine the chromosomal alterations of CGCCA, G-banding and spectral karyotyping studies were performed. The CGCCA cell line was transplanted into the nude mice for examining its tumorigenicity. 2-Deoxy-2-（18F）fluoro-D- glucose （FDG） autoradiography was also performed to evaluate the FDG uptake of the tumor xenograft. RESULTS： The doubling time for the CGCCA cell line was 32 h. After transplantation into nude mice, FDG autoradiography showed that the tumors formed at the cell transplantation site had a latency period of 4-6 wk with high FDG uptake excluding necrosis tissue. Moreover, immunohistochemical staining revealed prominent cytoplasmic expression of c-erb-B2, CK19, c-Met, COX-n, EGFR, MUC4, and a negative expression of K-ras. All data confirmed the phenotypic features of the CGCCA cell line coincide with the xenograft mice tumors, indicating cells containing the tumorigenicity of CCA originated from CCA. In addition, karyotypic band- ing analysis showed that the diploid （2n） cell status combines with ring and giant rod marker chromosomes in these clones; either both types simultaneously appeared or only one type of marker chromosome in a pair appeared in a cell. The major materials contained in the marker chromosome were primarily identified from chromosome 4. CONCLUSION： The current CGCCA cell line may be used as a non-K-ras effect CCA model and to obtain information and reveal novel pathways for CCA. Further applications regarding tumor markers or therapeutic targeting of CCA should be addressed accordingly.展开更多
BACKGROUND Simulated microgravity environment can lead to gastrointestinal motility disturbance.The pathogenesis of gastrointestinal motility disorders is closely related to the stem cell factor(SCF)/c-kit signaling p...BACKGROUND Simulated microgravity environment can lead to gastrointestinal motility disturbance.The pathogenesis of gastrointestinal motility disorders is closely related to the stem cell factor(SCF)/c-kit signaling pathway associated with intestinal flora and Cajal stromal cells.Moreover,intestinal flora can also affect the regulation of SCF/c-kit signaling pathway,thus affecting the expression of Cajal stromal cells.Cajal cells are the pacemakers of gastrointestinal motility.AIM To investigate the effects of Bifidobacterium lactis(B.lactis)BLa80 on the intestinal flora of rats in simulated microgravity and on the gastrointestinal motility-related SCF/c-kit pathway.METHODS The internationally recognized tail suspension animal model was used to simulate the microgravity environment,and 30 rats were randomly divided into control group,tail suspension group and drug administration tail suspension group with 10 rats in each group for a total of 28 days.The tail group was given B.lactis BLa80 by intragastric administration,and the other two groups were given water intragastric administration,the concentration of intragastric administration was 0.1 g/mL,and each rat was 1 mL/day.Hematoxylin&eosin staining was used to observe the histopathological changes in each segment of the intestine of each group,and the expression levels of SCF,c-kit,extracellular signal-regulated kinase(ERK)and p-ERK in the gastric antrum of each group were detected by Western blotting and PCR.The fecal flora and mucosal flora of rats in each group were detected by 16S rRNA.RESULTS Simulated microgravity resulted in severe exfoliation of villi of duodenum,jejunum and ileum in rats,marked damage,increased space between villi,loose arrangement,shortened columnar epithelium of colon,less folds,narrower mucosal thickness,reduced goblet cell number and crypts,and significant improvement after probiotic intervention.Simulated microgravity reduced the expressions of SCF and c-kit,and increased the expressions of ERK and P-ERK in the gastric antrum of rats.However,after probiotic intervention,the expressions of SCF and ckit were increased,while the expressions of ERK and P-ERK were decreased,with statistical significance(P<0.05).In addition,simulated microgravity can reduce the operational taxonomic unit(OTU)of the overall intestinal flora of rats,B.lactis BLa80 can increase the OTU of rats,simulated microgravity can reduce the overall richness and diversity of stool flora of rats,increase the abundance of firmicutes in stool flora of rats,and reduce the abundance of Bacteroides in stool flora of rats,most of which are mainly beneficial bacteria.Simulated microgravity can increase the overall richness and diversity of mucosal flora,increase the abundance of Bacteroides and Desulphurides in the rat mucosal flora,and decrease the abundance of firmicutes,most of which are proteobacteria.After probiotics intervention,the overall Bacteroidetes trend in simulated microgravity rats was increased.CONCLUSION B.lactis BLa80 can ameliorate intestinal mucosal injury,regulate intestinal flora,inhibit ERK expression,and activate the SCF/c-kit signaling pathway,which may have a facilitating effect on gastrointestinal motility in simulated microgravity rats.展开更多
BACKGROUND Magnetic compression anastomosis(MCA)offers a simple and reliable technique for inducing anastomoses at any point along the digestive tract.Evidence regarding whether the design of the MCA device influences...BACKGROUND Magnetic compression anastomosis(MCA)offers a simple and reliable technique for inducing anastomoses at any point along the digestive tract.Evidence regarding whether the design of the MCA device influences the anastomosis effect is lacking.AIM To investigate any difference in the side-to-side colonic anastomosis effect achieved with cylindrical vs circular ring magnets.METHODS We designed cylindrical and circular ring magnets suitable for side-to-side colonic anastomosis in rats.Thirty Sprague-Dawley rats were randomly divided into a cylindrical group,circular ring group,and cylindrical–circular ring group(n=10/group).Side-to-side colonic anastomosis was completed by transanal insertion of the magnets without incision of the colon.Operation time,perioperative complications,and magnet discharge time were recorded.Rats were euthanized 4 weeks postoperatively,and anastomotic specimens were obtained.The burst pressure and anastomotic diameter were measured sequentially,and anastomosis formation was observed by naked eye.Histological results were observed by light microscopy.RESULTS In all 30 rats,side-to-side colonic anastomosis was completed,for an operation success rate of 100%.No postoperative complications of bleeding and intestinal obstruction occurred,and the postoperative survival rate were 100%.The operation time,magnet discharge time,anastomotic bursting pressure,and anastomotic diameter did not differ significantly among the three designs(P>0.05).Healing was similar across the groups,with gross specimens showing good anastomotic healing and good mucosal continuity observed on histological analysis.CONCLUSION This study found no significant difference in the establishment of rat side-to-side colonic anastomosis with the use of cylindrical vs circular ring magnets.展开更多
Frozen shoulder(FS),also known as adhesive capsulitis,is a condition that causes contraction and stiffness of the shoulder joint capsule.The main symptoms are per-sistent shoulder pain and a limited range of motion in...Frozen shoulder(FS),also known as adhesive capsulitis,is a condition that causes contraction and stiffness of the shoulder joint capsule.The main symptoms are per-sistent shoulder pain and a limited range of motion in all directions.These symp-toms and poor prognosis affect people's physical health and quality of life.Currently,the specific mechanisms of FS remain unclear,and there is variability in treatment methods and their efficacy.Additionally,the early symptoms of FS are difficult to distinguish from those of other shoulder diseases,complicating early diagnosis and treatment.Therefore,it is necessary to develop and utilize animal models to under-stand the pathogenesis of FS and to explore treatment strategies,providing insights into the prevention and treatment of human FS.This paper reviews the rat models available for FS research,including external immobilization models,surgical internal immobilization models,injection modeling models,and endocrine modeling models.It introduces the basic procedures for these models and compares and analyzes the advantages,disadvantages,and applicability of each modeling method.Finally,our paper summarizes the common methods for evaluating FS rat models.展开更多
This paper introduces part of the content in the association standard,T/CAAM0002–2020 Nomenclature and Location of Acupuncture Points for Laboratory Animals Part 2:Rat.This standard was released by the China Associat...This paper introduces part of the content in the association standard,T/CAAM0002–2020 Nomenclature and Location of Acupuncture Points for Laboratory Animals Part 2:Rat.This standard was released by the China Association of Acupuncture and Moxibustion on May 15,2020,implemented on October 31,2020,and published by Standards Press of China.The standard was drafted by the Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,and the Nanjing University of Chinese Medicine.Principal draftsmen:Xiang-hong JING and Xing-bang HUA.Participating draftsmen:Wan-Zhu BAI,Bin XU,Dong-sheng XU,Yi GUO,Tie-ming MA,Xin-jun WANG,and Sheng-feng LU.展开更多
Background:Adenoid hypertrophy(AH)is a common pediatric disease that signifi-cantly impacts the growth and quality of life of children.However,there is no replica-ble and valid model for AH.Methods:An AH rat model was...Background:Adenoid hypertrophy(AH)is a common pediatric disease that signifi-cantly impacts the growth and quality of life of children.However,there is no replica-ble and valid model for AH.Methods:An AH rat model was developed via comprehensive allergic sensitization,chronic inflammation induction,and chronic intermittent hypoxia(CIH).The modeling process involved three steps:female Sprague-Dawley rats(aged 4-5 weeks)were used for modeling.Allergen sensitization was induced via intraperitoneal administra-tion and intranasal provocation using ovalbumin(OVA);chronic nasal inflammation was induced through intranasal lipopolysaccharide(LPS)administration for sustained nasal irritation;CIH akin to obstructive sleep apnea/hypopnea syndrome was induced using an animal hypoxia chamber.Postmodel establishment,behaviors,and histologi-cal changes in nasopharynx-associated lymphoid tissue(NALT)and nasal mucosa were assessed.Arterial blood gas analysis and quantification of serum and tissue levels of(interleukin)IL-4 and IL-13,OVA-specific immunoglobulin E(sIgE),eosinophil cationic protein(ECP),tumor necrosis factor(TNF-α),IL-17,and transforming growth factor(TGF)-βwere conducted for assessment.The treatment group received a combination of mometasone furoate and montelukast sodium for a week and then was evaluated.Results:Rats exhibited notable nasal symptoms and hypoxia after modeling.Histopathological analysis revealed NALT follicle hypertrophy and nasal mucosa in-flammatory cell infiltration.Elevated IL-4,IL-13,IL-17,OVA-sIgE,ECP,and TNF-αlev-els and reduced TGF-βlevels were observed in the serum and tissue of model-group rats.After a week of treatment,the treatment group exhibited symptom and inflam-matory factor improvement.Conclusion:The model effectively simulates AH symptoms and pathological changes.But it should be further validated for genetic,immunological,and hormonal back-grounds in the currently used and other strains and species.展开更多
Objective To explore the sequential effects of hypoxic exercising on miR-27/PPARγand lipid metabolism targetgene and protein expression levels in the obesity rats’liver.Methods 13-week-old male diet-induced obesity ...Objective To explore the sequential effects of hypoxic exercising on miR-27/PPARγand lipid metabolism targetgene and protein expression levels in the obesity rats’liver.Methods 13-week-old male diet-induced obesity rats were randomlydivided into three groups(n=10):normal oxygen concentration quiet group(N),hypoxia quiet group(H),hypoxic exercise group(HE).Exercise training on the horizontal animal treadmill for 1 h/d,5 d/week for a total of 4 week,and the intensity of horizontaltreadmill training was 20 m/min(hypoxic concentration was 13.6%).Comparison of the weights of perirenal fat and epididymal fat in rats across different groups and calculation of Lee’s index based on body weight and body length of rats in each group were done.And the serum concentrations of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),high-densitylipoprotein cholesterol(HDL-C)levels were detected.RT-PCR and Western Blot were used to detect the levels of miR-27,PPARγ,CYP7A1 and CD36.Results Hypoxic exercise decreased the expression levels of miR-27 in the obese rats’liver,however,theexpression level of PPARγwas gradually increased.The expression levels of miR-27 in HE group were significantly lower than Ngroup(P<0.05).The expression levels of PPARγmRNA in N group were significantly lower than H group(P<0.05),especially lowerthan HE group(P<0.01).The protein expression of PPARγprotein in N group was significantly lower than that other groups(P<0.01).The expression of lipid metabolism-related genes and proteins increased in the obese rats’liver.The expression of CYP7A1mRNA in N group was significantly lower than H group(P<0.05),especially lower than HE group(P<0.01).The expression ofCYP7A1 protein in the obese rats’liver in N group was extremely lower than H group and HE group(P<0.01).The proteinexpression of CD36 in N group was significantly lower than that in HE group(P<0.05).Hypoxia exercise improved the relatedphysiological and biochemical indexes of lipid metabolism disorder.The perirenal fat weight of obese rats in HE group wasextremely lower than N group and H group(P<0.01),and the perirenal fat weight in N group was significantly higher than H group(P<0.05).The epididymal fat weight in N group was significantly higher than H group(P<0.05),and extremely higher than HEgroup(P<0.01).The Lee’s index in HE group was extremely lower than N group and H group(P<0.01).The serum concentration ofTC in obese rats in HE group was extremely lower than N group and H group(P<0.01).The serum concentration of TG in HE groupwas extremely lower than N group and H group(P<0.01).The serum concentration of LDL-C in N group was extremely higher thanHE group(P<0.01).The serum concentration of HDL-C in N group was extremely lower than H group(P<0.01).Conclusion Hypoxiaand hypoxia exercise may negatively regulate the levels of PPARγby inhibiting miR-27 in the obese rats’liver,thereby affecting theexpression of downstream target genes CYP7A1 and CD36,and promoting cholesterol,fatty acid oxidation and HDL-C transport inthe liver,and ultimately the lipid levels in obese rats were improved.The effect of hypoxia exercise on improving blood lipid isbetter than simple hypoxia intervention.展开更多
This study aims to conduct a comparative study of two strains of laboratoryrats,hooded and Wistar,to select a suitable alternative to nude rats for ovariantissue xenotransplantation.The study investigated the effects ...This study aims to conduct a comparative study of two strains of laboratoryrats,hooded and Wistar,to select a suitable alternative to nude rats for ovariantissue xenotransplantation.The study investigated the effects of ovarian tissuetransplantation using three experimental groups:(1)the control group receivingvitrified-warmed human ovarian tissue,(2)the OTW(ovarian tissue transplantationin Wistar)group with human ovarian tissues in Wistar rats,and(3)the OTH(ovariantissue transplantation in hooded)group with ovarian tissues transplanted into hoodedrats.A total of 12 rats(6 each from the two transplantation groups)were used,withtissue samples implanted in the dorsal neck muscles.Outcomes were evaluatedusing histological analyses,including hematoxylin and eosin and Masson's trichromestaining,as well as immunohistochemical assessments of CD31 for angiogenesis,fibrosis,and necrosis.The OTH group exhibited more blood vessel numbers and theVegf gene expression than both the OTW and control groups,with significantly lowerlevels of fibrosis and necrosis compared to the OTW and control groups.The studyindicates that hooded rats could serve as a valuable alternative for specific researchwhen nude mice are not available.Their immune systems are more compatible withhuman ovarian tissue transplants than Wistar rats,and they exhibit calmer behaviors,making them better suited for laboratory work.展开更多
Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tari(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neu...Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tari(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neural circuit tracing.Anterograde labeling was achieved by injecting anterograde self-complementary adeno-associated viruses(scAAVs)into the IC.Retrograde tracing involved NTS injections of either retrograde scAAVs or FluoroGold(FG),combined with immunofluorescence histochemical staining to identify IC-originating projection neurons.For postsynaptic neurochemical phenotype characterization,IC was injected with AAV2/1-CaMKII-Cre,while a mixture of AAV2/9-Syn-DIO-mCherry and AAV2/9-VGAT1-EGFP was injected into the NTS.The rats were allowed to survive for one week following scAAVs or FG injection or four weeks after recombinase-dependent systems injection.Then the rats were sacrificed,and serial brain sections were prepared for immunofluorescence histochemical staining(brain section containing FG)and subsequent fluorescence/confocal microscopic analysis.Results:(1)Anterograde viral tracing re-vealed dense axonal terminals from the IC projecting to the medial subnucleus of the NTS,while retrograde tracing re-vealed that IC neurons projecting to the NTS were predominantly localized within the dysgranular layer;(2)IC-NTS projection neurons were exclusive glutamatergic(100%,n=3);(3)NTS neurons receiving IC inputs were mainly lo-calized in the medial subnucleus,and were predominantly GABAergic(79.8±3.2%,n=3).Conclusion:The pres-ent results indicate that a descending pathway from excitatory neurons of the IC terminates onto inhibitory neurons of the NTS,which might represent a potential neuromodulatory target for visceral pain disorders.展开更多
Background:Scientific animal models are indispensable for studying trauma repair.This work aimed at establishing a more scientific rat trauma model by studying different rat trauma models caused by different trauma nu...Background:Scientific animal models are indispensable for studying trauma repair.This work aimed at establishing a more scientific rat trauma model by studying different rat trauma models caused by different trauma numbers,locations,and trauma attachment tension unloaders and rat age.Methods:A four-trauma self-upper,lower,left and right control model;a two-trauma self-trauma bare and ring control model;and a young and old rat trauma model were created to evaluate the condition of these traumas.Results:In the four-trauma self-control model,the healing status of the upper proximal cephalic trauma was better than that of the lower proximal caudal trauma,whereas there was no significant difference between the left and right trauma.The healing rate and postwound condition of the trauma with a ring control in the two-trauma model were better than those of the bare side.The healing speed of the old rats was slower,and the amount of extracellular matrix in the subcutaneous tissue after healing was significantly lower than that of the young rats.Conclusion:The double trauma with a ring is a more scientific and reasonable experimental model.There is a significant difference between young and old rats in the wound healing process.Therefore,the appropriate age of the rats should be selected according to the main age range of the patients with similar conditions in the clinical setting being mimicked.展开更多
[Objectives]To investigate the effects of different doses of chili pepper on inflammatory response factors in rats.[Methods]Sixty male SD rats were randomly divided into blank control group,high-dose chili pepper grou...[Objectives]To investigate the effects of different doses of chili pepper on inflammatory response factors in rats.[Methods]Sixty male SD rats were randomly divided into blank control group,high-dose chili pepper group,medium-dose chili pepper group,and low-dose chili pepper group.Chili pepper aqueous solution was administered by gavage for three months.One day after the last administration,serum and cerebrospinal fluid were collected to measure the concentrations of hs-CRP and IL-6 in serum,and PCT in cerebrospinal fluid.[Results]Compared with the blank control group,the high-dose,medium-dose,and low-dose chili pepper groups showed significant increases in hs-CRP,IL-6,and PCT levels(P<0.05).[Conclusions]Different doses of chili pepper exerted varying effects on inflammatory factors in rats.展开更多
This study aimed to assess the therapeutic potential of Anemarrhenae Rhizoma(AR)in osteoporotic rats and to elucidate the metabolic pathways involved in AR’s role in alleviating osteoporosis(OP).OP was induced in rat...This study aimed to assess the therapeutic potential of Anemarrhenae Rhizoma(AR)in osteoporotic rats and to elucidate the metabolic pathways involved in AR’s role in alleviating osteoporosis(OP).OP was induced in rats through ovariectomy(OVX),followed by oral administration of either high or low doses of AR,as well as estradiol valerate,over a 14-week period.Micro-computed tomography(Micro-CT)was employed to examine the femur tissue morphology,while enzyme-linked immunosorbent assay(ELISA)was adopted to measure serum levels of PINP and CTX-I to evaluate AR’s efficacy in treating OP.Additionally,metabolomic profiling of femur tissues was conducted using gas chromatography-mass spectrometry(GC-MS).The bioactive components of AR,along with its therapeutic targets for OP,were identified through UPLC-MS/MS and online database searches,and metabolic networks were established by integrating differential metabolites and potential targets.Furthermore,Western blotting analysis confirmed key molecular targets.The findings revealed that AR treatment significantly mitigated OVX-induced OP in rats.Metabolomic analysis indicated that AR exerted its effects by modulating the levels of 10 key metabolites(such as linoleic acid and inositol)and influencing five crucial metabolic pathways,including linoleic acid metabolism and the phosphoinositide signaling system.Among these,the linoleic acid metabolic pathway emerged as a pivotal focus for further investigation based on the constructed interaction network of differential metabolites and targets.Western blotting analysis demonstrated that AR reversed the up-regulation of CYP1A2 and CYP2C9,two targets associated with the linoleic acid metabolic pathwa y,in OP rats.In conclusion,AR appeared to ameliorate OP by modulating metabolite levels in OVX rats,with its mechanism of action likely centered on regulating the linoleic acid metabolic pathway.展开更多
The COVID-19 pandemic posed a challenge for clinical management of a new lung disease that was characterized by inflammation,endothelial cell dysfunction,and thrombosis,which occur after the replication phase of infec...The COVID-19 pandemic posed a challenge for clinical management of a new lung disease that was characterized by inflammation,endothelial cell dysfunction,and thrombosis,which occur after the replication phase of infection of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).There are many laboratory models of active SARS-CoV-2 infection in mice,reflecting an acute lung injury in an otherwise healthy animal,but there is a lack of accurate animal models of the postviral inflammatory phase of the COVID-19 lung reflecting severe disease.The monocrotaline(MCT)-treated rat is a widely used laboratory model of pulmonary hypertension(PH).Not often discussed,however,are the observed changes in inflammation,edema,fibrosis,and microthrombosis in the lung prior to PH.At the cellular level,there is loss of pneumocytes and endotheliopathy,and at the molecular level the MCT rat lung is characterized by a proinflammatory cytokine profile,namely elevated interleukin 6,transforming growth factorβand tumor necrosis factor,M1 macrophage phenotype,and dysregulation of the angiotensin converting enzyme(ACE)/ACE2 balance.The systems-level pathophysiology of the MCT-treated rat includes progressive cardiopulmonary dysfunction.The MCT-treated rat clearly differs from the COVID-19 lung in terms of the triggers for pathology,but there are many parallels apparent in both the MCT-treated rat and the COVID-19 lung.The MCT-treated rat lung as a model of the COVID-19 lung may provide an in-depth understanding of the factors that drive the lung to more severe pathology,treatments that benefit lung recovery,or the factors that prove a useful research platform for future emerging respiratory threats of similar pathology.展开更多
Background:C1QL3 is widely expressed in the brain and is specifically produced by a subset of excitatory neurons.However,its function is still not clear.We established C1ql3-deficient rats to investigate the role of C...Background:C1QL3 is widely expressed in the brain and is specifically produced by a subset of excitatory neurons.However,its function is still not clear.We established C1ql3-deficient rats to investigate the role of C1QL3 in the brain.Methods:C1ql3 knockout(KO)rats were generated using CRISPR/Cas9.C1ql3 KO was determined by polymerase chain reaction(PCR),DNA sequencing,and western blot-ting.Microglia morphology and cytokine expression with or without lipopolysaccha-ride(LPS)stimulus were analyzed using immunohistochemistry and real-time PCR.The brain structure changes in KO rats were examined using magnetic resonance imaging.Neuronal architecture alteration was analyzed by performing Golgi staining.Behavior was evaluated using the open field test,Morris water maze test,and Y maze test.Results:C1ql3 KO significantly increased the number of ramified microglia and decreased the number of hypertrophic microglia,whereas C1ql3 KO did not in-fluence the expression of pro-inflammatory factors and anti-inflammatory factors except IL-10.C1ql3 KO brains had more amoeboid microglia types and higher Arg-1 expression compared with the WT rats after LPS stimulation.The brain weights and HPC sizes of C1ql3 KO rats did not differ from WT rats.C1ql3 KO damaged neuronal integrity including neuron dendritic arbors and spine density.C1ql3 KO rats demonstrated an increase in spontaneous activity and an impairment in short working memory.Conclusions:C1ql3 KO not only interrupts the neuronal integrity but also affects the microglial activation,resulting in hyperactive behavior and impaired short memory in rats,which highlights the role of C1QL3 in the regulation of structure and function of both neuronal and microglial cells.展开更多
Background:The Cre/loxP system is most popular in mice,but its application in rats has largely lagged far behind.The rat is vital laboratory animal,especially in toxicological and neurological studies.Generating genet...Background:The Cre/loxP system is most popular in mice,but its application in rats has largely lagged far behind.The rat is vital laboratory animal,especially in toxicological and neurological studies.Generating genetic tools to manipulate neurons in rats could benefit neurological research.Methods:Using the CRISPR/Cas9 system,we inserted a Cre cassette into endogenous Thy1 and NeuN loci.Thy1-Cre rats featured a downstream P2A-linked insertion,while NeuN-Cre was inserted at the transcriptional start site.The Cre activity was assessed by crossing with a Cre reporter(Rosa26 imCherry)rat and through analyzing mCherry expression patterns.The specificity of cell type was further confirmed by immunofluorescence with NeuN antibody.Phenotypic consequences were assessed by crossing with ND1^(LSL) rats to deplete ND1,followed by monitoring weight/survival and conducting motor function tests.Results:We generated two neuron-specific rats(Thy1-Cre and NeuN-Cre),which exhibited high neuron-specific Cre expression in brain and spinal cord with minor leakage in other tissues.Thy1-Cre showed minor leakage in spleen,lung and kidney while NeuN-Cre showed minor leakage in spleen and kidney.ND1^(Thy1-Cre) and ND1^(NeuN-Cre) rats both showed decreased body weights and survival times.The ND1^(NeuN-Cre) rats died within two weeks,while ND1^(Thy1-Cre) rats lived longer with impaired motor function.Conclusions:We successfully generated two neuron-specific NeuN-Cre and Thy1-Cre rats,and systemically analyzed their expression pattern.展开更多
Humans and animals have a fundamental ability to use experiences and environmental information to organize behavior.It often happens that humans and animals make decisions and prepare actions under uncertain situation...Humans and animals have a fundamental ability to use experiences and environmental information to organize behavior.It often happens that humans and animals make decisions and prepare actions under uncertain situations.Uncertainty would significantly affect the state of animals’minds,but may not be reflected in behavior.How to“read animals’mind state”under different situations is a challenge.Here,we report that neuronal activity in the medial prefrontal cortex(mPFC)of rats can reflect the environmental uncertainty when the task situation changes from certain to uncertain.Rats were trained to perform behavioral tasks under certain and uncertain situations.Under certain situations,rats were required to simply repeat two nose-poking actions that each triggered short auditory tone feedback(single-task situation).Whereas under the uncertain situation,the feedback could randomly be either the previous tone or a short musical rhythm.No additional action was required upon the music feedback,and the same secondary nose-poking action was required upon the tone feedback(dual-task situation);therefore,the coming task was uncertain before action initiation.We recorded single-unit activity from the mPFC when the rats were performing the tasks.We found that in the dual task,when uncertainty was introduced,many mPFC neurons were actively engaged in dealing with the uncertainty before the task initiation,suggesting that the rats could be aware of the task situation change and encode the information in the mPFC before the action of task initiation.展开更多
Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed ra...Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed rat resource has rapidly increased,but the liver-specific Cre rat lines for studying liver diseases and interested genes are limited,especially in a spatially and temporally restricted manner.Methods:RNA sequencing and real-time polymerase chain reaction(PCR)were used to screen and confirm the presence of liver-specific genes.Apoa4-Cre rats and Cyp2c11-Cre rats were produced by CRISPR/Cas9 knockin.Rosa26-imCherry rats were employed to hybridize with the Cre rats to obtain the Apoa4-Cre/Rosa26-imCherry and Cyp2c11-Cre/Rosa26-imCherry rats.The temporal and spatial patterns of Cre expression were determined by the observation of red fluorescence on tis-sue sections.Hematoxylin-eosin stain was used to evaluate the liver histopathologic changes.The blood biochemical analysis of several liver enzymes and liver lipid profile was performed to evaluate the liver function of Cre rats.Results:Apoa4 and Cyp2c11 were identified as two liver-specific genes.Apoa4-Cre and Cyp2c11-Cre rats were produced and hybridized with Rosa26-imCherry rats.The red fluorescence indicated that the Cre recombinases were specially expressed in the juvenile and adult liver and not in other organs of two hybridized rats.All the blood biochemical parameters except low-density lipoprotein(LDL)did not change signifi-cantly in the Cre rats.No histological alterations were detected in the livers of the Cre rats.Conclusions:Liver-specific Apoa4-Cre and Cyp2c11-Cre rats have been established successfully and could be used to study gene knockout,specifically in juvenile and adult liver.展开更多
Although Steen's solution plays a crucial role in maintaining vascular oncotic pressure and reducing pulmonary edema due to its high concentration of human serum albumin during ex vivo lung perfusion(EVLP),it incu...Although Steen's solution plays a crucial role in maintaining vascular oncotic pressure and reducing pulmonary edema due to its high concentration of human serum albumin during ex vivo lung perfusion(EVLP),it incurs high costs and potential immune responses in rat models.In our preliminary research,donor lungs perfused with Steen's solution exhibited significant lung injury post transplantation,likely due to a strong immune response elicited by the high concentration of xenogenic proteins upon reperfusion.This study explores a low-cost and efficient perfusate composition for use in rat EVLP and lung transplant models.The modified perfusate employed in this study uses artificial colloids as the primary component,with an albumin concentration of only 2%,demonstrating comparable physiological parameters,metabolic levels,and pathological changes to those of Steen's solution during EVLP.Moreover,the modified composition exhibited good biocompatibility in transplantation,with minimal lung injury.In summary,the proposed perfusate offers a safer and more economical alternative for establishing rat EVLP and lung transplant models,facilitating the advancement of related research.展开更多
基金financially assisted to Timanshi Chansoriya of the fellowship grant of UGC-NFPwD,New Delhi sanctioned vide UGC F.No.01-01/2019-Sch dated 06.11.2020(Serial No.352 ID:NFPWD-2018-20-UTP-6522).
文摘Objective:To investigate the sterilizing potential of zinc gluconate,calcium chloride(CaCl_(2))and cadmium chloride(CdCl_(2))following a single intratesticular administration in adult male rats.Methods:60 adult male Wistar albino rats(Rattus norvegicus)weighing 160-200 g and aged 5-6 months randomly received a single intratesticular injection of normal saline(Group A),zinc gluconate 13.3 mg/mL plus L-Arginine(Group B),20%CaCl_(2)(Group C),and CdCl_(2)0.5 mg/kg body weight plus ethylenediaminetetraacetic acid(EDTA)(Group D),respectively,along the entire route from the caudoventral aspect of each testis.They were euthanized up to 180 days to evaluate reproductive tract toxicology.Results:The reproductive organ weights were markedly reduced,with testes severely atrophied in group B,pea-sized and stony hard in group C,and moderately reduced in group D.Azoospermia was evident in groups B and C,while sperm concentration was reduced to<1 million/mL with zero sperm motility in group D.Rats of groups B and C failed to show mounting and copulatory behaviour.A completely disorganized mesh of cellular elements was observed in the seminiferous tubules of group B,while pyknotic germ cell and arrest of spermatogenesis,exfoliated germ cells,occasional syncytial bodies and smaller Leydig cells were evident in groups C and D.Significantly reduced testosterone levels,increased luteinizing hormone(LH)and follicle-stimulating hormone(FSH)levels,returned to normal after 90 days in group D.Conclusions:Zinc gluconate and CaCl_(2)injections severely affected the reproductive organs and libido and rats treated with CdCl_(2)exhibited diminished spermatogenesis with normal libido.Thus,the need-based selection of intratesticular agents should consider their distinct effects on spermatogenesis,libido,and hormonal balance for achieving targeted sterilization outcomes.
基金supported by the National Natural Science Foundation of China,Nos.82271132(to YL),82101167(to BB)the Natural Science Foundation of Chongqing,Nos.CSTB2022NSCQ-MSX0020(to BB),cstc2019jcyj-msxmX0473(to FC).
文摘Our previous study demonstrated that combined transplantation of bone marrow mesenchymal stem cells and retinal progenitor cells in rats has therapeutic effects on retinal degeneration that are superior to transplantation of retinal progenitor cells alone.Bone marrow mesenchymal stem cells regulate and interact with various cells in the retinal microenvironment by secreting neurotrophic factors and extracellular vesicles.Small extracellular vesicles derived from bone marrow mesenchymal stem cells,which offer low immunogenicity,minimal tumorigenic risk,and ease of transportation,have been utilized in the treatment of various neurological diseases.These vesicles exhibit various activities,including anti-inflammatory actions,promotion of tissue repair,and immune regulation.Therefore,novel strategies using human retinal progenitor cells combined with bone marrow mesenchymal stem cell-derived small extracellular vesicles may represent an innovation in stem cell therapy for retinal degeneration.In this study,we developed such an approach utilizing retinal progenitor cells combined with bone marrow mesenchymal stem cell-derived small extracellular vesicles to treat retinal degeneration in Royal College of Surgeons rats,a genetic model of retinal degeneration.Our findings revealed that the combination of bone marrow mesenchymal stem cell-derived small extracellular vesicles and retinal progenitor cells significantly improved visual function in these rats.The addition of bone marrow mesenchymal stem cell-derived small extracellular vesicles as adjuvants to stem cell transplantation with retinal progenitor cells enhanced the survival,migration,and differentiation of the exogenous retinal progenitor cells.Concurrently,these small extracellular vesicles inhibited the activation of regional microglia,promoted the migration of transplanted retinal progenitor cells to the inner nuclear layer of the retina,and facilitated their differentiation into photoreceptors and bipolar cells.These findings suggest that bone marrow mesenchymal stem cell-derived small extracellular vesicles potentiate the therapeutic efficacy of retinal progenitor cells in retinal degeneration by promoting their survival and differentiation.
文摘AIM： TO characterize a culture model of rat CCA cells, which were derived from a transplantable TTA-induced CCA and designated as Chang Gung CCA （CGCCA）. METHODS： The CGCCA cells were cultured at in vitro passage 12 times on a culture dish in DMEM medium. To measure the doubling time, 103 cells were plated in a 96-well plate containing the growth medium. The cells were harvested 4 to 10 d after seeding, and a standard MTT assay was used to measure the growth. The phenotype of CACCA cell and xenograft was determined by immunohistochemical study. We also determine the chromosomal alterations of CGCCA, G-banding and spectral karyotyping studies were performed. The CGCCA cell line was transplanted into the nude mice for examining its tumorigenicity. 2-Deoxy-2-（18F）fluoro-D- glucose （FDG） autoradiography was also performed to evaluate the FDG uptake of the tumor xenograft. RESULTS： The doubling time for the CGCCA cell line was 32 h. After transplantation into nude mice, FDG autoradiography showed that the tumors formed at the cell transplantation site had a latency period of 4-6 wk with high FDG uptake excluding necrosis tissue. Moreover, immunohistochemical staining revealed prominent cytoplasmic expression of c-erb-B2, CK19, c-Met, COX-n, EGFR, MUC4, and a negative expression of K-ras. All data confirmed the phenotypic features of the CGCCA cell line coincide with the xenograft mice tumors, indicating cells containing the tumorigenicity of CCA originated from CCA. In addition, karyotypic band- ing analysis showed that the diploid （2n） cell status combines with ring and giant rod marker chromosomes in these clones; either both types simultaneously appeared or only one type of marker chromosome in a pair appeared in a cell. The major materials contained in the marker chromosome were primarily identified from chromosome 4. CONCLUSION： The current CGCCA cell line may be used as a non-K-ras effect CCA model and to obtain information and reveal novel pathways for CCA. Further applications regarding tumor markers or therapeutic targeting of CCA should be addressed accordingly.
文摘BACKGROUND Simulated microgravity environment can lead to gastrointestinal motility disturbance.The pathogenesis of gastrointestinal motility disorders is closely related to the stem cell factor(SCF)/c-kit signaling pathway associated with intestinal flora and Cajal stromal cells.Moreover,intestinal flora can also affect the regulation of SCF/c-kit signaling pathway,thus affecting the expression of Cajal stromal cells.Cajal cells are the pacemakers of gastrointestinal motility.AIM To investigate the effects of Bifidobacterium lactis(B.lactis)BLa80 on the intestinal flora of rats in simulated microgravity and on the gastrointestinal motility-related SCF/c-kit pathway.METHODS The internationally recognized tail suspension animal model was used to simulate the microgravity environment,and 30 rats were randomly divided into control group,tail suspension group and drug administration tail suspension group with 10 rats in each group for a total of 28 days.The tail group was given B.lactis BLa80 by intragastric administration,and the other two groups were given water intragastric administration,the concentration of intragastric administration was 0.1 g/mL,and each rat was 1 mL/day.Hematoxylin&eosin staining was used to observe the histopathological changes in each segment of the intestine of each group,and the expression levels of SCF,c-kit,extracellular signal-regulated kinase(ERK)and p-ERK in the gastric antrum of each group were detected by Western blotting and PCR.The fecal flora and mucosal flora of rats in each group were detected by 16S rRNA.RESULTS Simulated microgravity resulted in severe exfoliation of villi of duodenum,jejunum and ileum in rats,marked damage,increased space between villi,loose arrangement,shortened columnar epithelium of colon,less folds,narrower mucosal thickness,reduced goblet cell number and crypts,and significant improvement after probiotic intervention.Simulated microgravity reduced the expressions of SCF and c-kit,and increased the expressions of ERK and P-ERK in the gastric antrum of rats.However,after probiotic intervention,the expressions of SCF and ckit were increased,while the expressions of ERK and P-ERK were decreased,with statistical significance(P<0.05).In addition,simulated microgravity can reduce the operational taxonomic unit(OTU)of the overall intestinal flora of rats,B.lactis BLa80 can increase the OTU of rats,simulated microgravity can reduce the overall richness and diversity of stool flora of rats,increase the abundance of firmicutes in stool flora of rats,and reduce the abundance of Bacteroides in stool flora of rats,most of which are mainly beneficial bacteria.Simulated microgravity can increase the overall richness and diversity of mucosal flora,increase the abundance of Bacteroides and Desulphurides in the rat mucosal flora,and decrease the abundance of firmicutes,most of which are proteobacteria.After probiotics intervention,the overall Bacteroidetes trend in simulated microgravity rats was increased.CONCLUSION B.lactis BLa80 can ameliorate intestinal mucosal injury,regulate intestinal flora,inhibit ERK expression,and activate the SCF/c-kit signaling pathway,which may have a facilitating effect on gastrointestinal motility in simulated microgravity rats.
基金Supported by the Key Research and Development Program of Shaanxi,No.2024SF-YBXM-447the Institutional Foundation of The First Affiliated Hospital of Xi’an Jiaotong University,No.2022MS-07the Fundamental Research Funds for the Central Universities,No.xzy022023068.
文摘BACKGROUND Magnetic compression anastomosis(MCA)offers a simple and reliable technique for inducing anastomoses at any point along the digestive tract.Evidence regarding whether the design of the MCA device influences the anastomosis effect is lacking.AIM To investigate any difference in the side-to-side colonic anastomosis effect achieved with cylindrical vs circular ring magnets.METHODS We designed cylindrical and circular ring magnets suitable for side-to-side colonic anastomosis in rats.Thirty Sprague-Dawley rats were randomly divided into a cylindrical group,circular ring group,and cylindrical–circular ring group(n=10/group).Side-to-side colonic anastomosis was completed by transanal insertion of the magnets without incision of the colon.Operation time,perioperative complications,and magnet discharge time were recorded.Rats were euthanized 4 weeks postoperatively,and anastomotic specimens were obtained.The burst pressure and anastomotic diameter were measured sequentially,and anastomosis formation was observed by naked eye.Histological results were observed by light microscopy.RESULTS In all 30 rats,side-to-side colonic anastomosis was completed,for an operation success rate of 100%.No postoperative complications of bleeding and intestinal obstruction occurred,and the postoperative survival rate were 100%.The operation time,magnet discharge time,anastomotic bursting pressure,and anastomotic diameter did not differ significantly among the three designs(P>0.05).Healing was similar across the groups,with gross specimens showing good anastomotic healing and good mucosal continuity observed on histological analysis.CONCLUSION This study found no significant difference in the establishment of rat side-to-side colonic anastomosis with the use of cylindrical vs circular ring magnets.
基金National Key R&D Program of China,Grant/Award Number:2021YFC2502100,2023YFC3603404 and 2019YFA0111900The National Natural Science Foundation of China,Grant/Award Number:82072506,82272611 and 92268115+7 种基金The Hunan Provincial Science Fund for Distinguished Young Scholars,Grant/Award Number:2024JJ2089The Hunan Young Talents of Science and Technology,Grant/Award Number:2021RC3025The Provincial Clinical Medical Technology Innovation Project of Hunan,Grant/Award Number:2023SK2024 and 2020SK53709The Provincial Natural Science Foundation of Hunan,Grant/Award Number:2020JJ3060The National Natural Science Foundation of Hunan Province,Grant/Award Number:2023JJ30949The National Clinical Research Center for Geriatric Disorders,Xiangya Hospital,Grant/Award Number:2021KFJJ02 and 2021LNJJ05The Hunan Provincial Innovation Foundation for Postgraduate,Grant/Award Number:CX20230308 and CX20230312The Independent Exploration and Innovation Project for Postgraduate Students of Central South University,Grant/Award Number:2024ZZTS0163。
文摘Frozen shoulder(FS),also known as adhesive capsulitis,is a condition that causes contraction and stiffness of the shoulder joint capsule.The main symptoms are per-sistent shoulder pain and a limited range of motion in all directions.These symp-toms and poor prognosis affect people's physical health and quality of life.Currently,the specific mechanisms of FS remain unclear,and there is variability in treatment methods and their efficacy.Additionally,the early symptoms of FS are difficult to distinguish from those of other shoulder diseases,complicating early diagnosis and treatment.Therefore,it is necessary to develop and utilize animal models to under-stand the pathogenesis of FS and to explore treatment strategies,providing insights into the prevention and treatment of human FS.This paper reviews the rat models available for FS research,including external immobilization models,surgical internal immobilization models,injection modeling models,and endocrine modeling models.It introduces the basic procedures for these models and compares and analyzes the advantages,disadvantages,and applicability of each modeling method.Finally,our paper summarizes the common methods for evaluating FS rat models.
文摘This paper introduces part of the content in the association standard,T/CAAM0002–2020 Nomenclature and Location of Acupuncture Points for Laboratory Animals Part 2:Rat.This standard was released by the China Association of Acupuncture and Moxibustion on May 15,2020,implemented on October 31,2020,and published by Standards Press of China.The standard was drafted by the Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,and the Nanjing University of Chinese Medicine.Principal draftsmen:Xiang-hong JING and Xing-bang HUA.Participating draftsmen:Wan-Zhu BAI,Bin XU,Dong-sheng XU,Yi GUO,Tie-ming MA,Xin-jun WANG,and Sheng-feng LU.
基金This work was financially supported by the National Natural Science Foundation of China(grant number:8217150152)the Clinical Science and Technology Innovation Project of Shanghai Shenkang Hospital Development Center(grant number:SHDC12021102)the Shanghai Three-Year Action Plan to Further Accelerate the Development of Traditional Chinese Medicine Inheritance and Innovation(grant number:ZY(2021-2023)-0209-05).
文摘Background:Adenoid hypertrophy(AH)is a common pediatric disease that signifi-cantly impacts the growth and quality of life of children.However,there is no replica-ble and valid model for AH.Methods:An AH rat model was developed via comprehensive allergic sensitization,chronic inflammation induction,and chronic intermittent hypoxia(CIH).The modeling process involved three steps:female Sprague-Dawley rats(aged 4-5 weeks)were used for modeling.Allergen sensitization was induced via intraperitoneal administra-tion and intranasal provocation using ovalbumin(OVA);chronic nasal inflammation was induced through intranasal lipopolysaccharide(LPS)administration for sustained nasal irritation;CIH akin to obstructive sleep apnea/hypopnea syndrome was induced using an animal hypoxia chamber.Postmodel establishment,behaviors,and histologi-cal changes in nasopharynx-associated lymphoid tissue(NALT)and nasal mucosa were assessed.Arterial blood gas analysis and quantification of serum and tissue levels of(interleukin)IL-4 and IL-13,OVA-specific immunoglobulin E(sIgE),eosinophil cationic protein(ECP),tumor necrosis factor(TNF-α),IL-17,and transforming growth factor(TGF)-βwere conducted for assessment.The treatment group received a combination of mometasone furoate and montelukast sodium for a week and then was evaluated.Results:Rats exhibited notable nasal symptoms and hypoxia after modeling.Histopathological analysis revealed NALT follicle hypertrophy and nasal mucosa in-flammatory cell infiltration.Elevated IL-4,IL-13,IL-17,OVA-sIgE,ECP,and TNF-αlev-els and reduced TGF-βlevels were observed in the serum and tissue of model-group rats.After a week of treatment,the treatment group exhibited symptom and inflam-matory factor improvement.Conclusion:The model effectively simulates AH symptoms and pathological changes.But it should be further validated for genetic,immunological,and hormonal back-grounds in the currently used and other strains and species.
文摘Objective To explore the sequential effects of hypoxic exercising on miR-27/PPARγand lipid metabolism targetgene and protein expression levels in the obesity rats’liver.Methods 13-week-old male diet-induced obesity rats were randomlydivided into three groups(n=10):normal oxygen concentration quiet group(N),hypoxia quiet group(H),hypoxic exercise group(HE).Exercise training on the horizontal animal treadmill for 1 h/d,5 d/week for a total of 4 week,and the intensity of horizontaltreadmill training was 20 m/min(hypoxic concentration was 13.6%).Comparison of the weights of perirenal fat and epididymal fat in rats across different groups and calculation of Lee’s index based on body weight and body length of rats in each group were done.And the serum concentrations of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),high-densitylipoprotein cholesterol(HDL-C)levels were detected.RT-PCR and Western Blot were used to detect the levels of miR-27,PPARγ,CYP7A1 and CD36.Results Hypoxic exercise decreased the expression levels of miR-27 in the obese rats’liver,however,theexpression level of PPARγwas gradually increased.The expression levels of miR-27 in HE group were significantly lower than Ngroup(P<0.05).The expression levels of PPARγmRNA in N group were significantly lower than H group(P<0.05),especially lowerthan HE group(P<0.01).The protein expression of PPARγprotein in N group was significantly lower than that other groups(P<0.01).The expression of lipid metabolism-related genes and proteins increased in the obese rats’liver.The expression of CYP7A1mRNA in N group was significantly lower than H group(P<0.05),especially lower than HE group(P<0.01).The expression ofCYP7A1 protein in the obese rats’liver in N group was extremely lower than H group and HE group(P<0.01).The proteinexpression of CD36 in N group was significantly lower than that in HE group(P<0.05).Hypoxia exercise improved the relatedphysiological and biochemical indexes of lipid metabolism disorder.The perirenal fat weight of obese rats in HE group wasextremely lower than N group and H group(P<0.01),and the perirenal fat weight in N group was significantly higher than H group(P<0.05).The epididymal fat weight in N group was significantly higher than H group(P<0.05),and extremely higher than HEgroup(P<0.01).The Lee’s index in HE group was extremely lower than N group and H group(P<0.01).The serum concentration ofTC in obese rats in HE group was extremely lower than N group and H group(P<0.01).The serum concentration of TG in HE groupwas extremely lower than N group and H group(P<0.01).The serum concentration of LDL-C in N group was extremely higher thanHE group(P<0.01).The serum concentration of HDL-C in N group was extremely lower than H group(P<0.01).Conclusion Hypoxiaand hypoxia exercise may negatively regulate the levels of PPARγby inhibiting miR-27 in the obese rats’liver,thereby affecting theexpression of downstream target genes CYP7A1 and CD36,and promoting cholesterol,fatty acid oxidation and HDL-C transport inthe liver,and ultimately the lipid levels in obese rats were improved.The effect of hypoxia exercise on improving blood lipid isbetter than simple hypoxia intervention.
文摘This study aims to conduct a comparative study of two strains of laboratoryrats,hooded and Wistar,to select a suitable alternative to nude rats for ovariantissue xenotransplantation.The study investigated the effects of ovarian tissuetransplantation using three experimental groups:(1)the control group receivingvitrified-warmed human ovarian tissue,(2)the OTW(ovarian tissue transplantationin Wistar)group with human ovarian tissues in Wistar rats,and(3)the OTH(ovariantissue transplantation in hooded)group with ovarian tissues transplanted into hoodedrats.A total of 12 rats(6 each from the two transplantation groups)were used,withtissue samples implanted in the dorsal neck muscles.Outcomes were evaluatedusing histological analyses,including hematoxylin and eosin and Masson's trichromestaining,as well as immunohistochemical assessments of CD31 for angiogenesis,fibrosis,and necrosis.The OTH group exhibited more blood vessel numbers and theVegf gene expression than both the OTW and control groups,with significantly lowerlevels of fibrosis and necrosis compared to the OTW and control groups.The studyindicates that hooded rats could serve as a valuable alternative for specific researchwhen nude mice are not available.Their immune systems are more compatible withhuman ovarian tissue transplants than Wistar rats,and they exhibit calmer behaviors,making them better suited for laboratory work.
文摘Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tari(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neural circuit tracing.Anterograde labeling was achieved by injecting anterograde self-complementary adeno-associated viruses(scAAVs)into the IC.Retrograde tracing involved NTS injections of either retrograde scAAVs or FluoroGold(FG),combined with immunofluorescence histochemical staining to identify IC-originating projection neurons.For postsynaptic neurochemical phenotype characterization,IC was injected with AAV2/1-CaMKII-Cre,while a mixture of AAV2/9-Syn-DIO-mCherry and AAV2/9-VGAT1-EGFP was injected into the NTS.The rats were allowed to survive for one week following scAAVs or FG injection or four weeks after recombinase-dependent systems injection.Then the rats were sacrificed,and serial brain sections were prepared for immunofluorescence histochemical staining(brain section containing FG)and subsequent fluorescence/confocal microscopic analysis.Results:(1)Anterograde viral tracing re-vealed dense axonal terminals from the IC projecting to the medial subnucleus of the NTS,while retrograde tracing re-vealed that IC neurons projecting to the NTS were predominantly localized within the dysgranular layer;(2)IC-NTS projection neurons were exclusive glutamatergic(100%,n=3);(3)NTS neurons receiving IC inputs were mainly lo-calized in the medial subnucleus,and were predominantly GABAergic(79.8±3.2%,n=3).Conclusion:The pres-ent results indicate that a descending pathway from excitatory neurons of the IC terminates onto inhibitory neurons of the NTS,which might represent a potential neuromodulatory target for visceral pain disorders.
基金Shandong Provincial Natural Science Foundation,Grant/Award Number:ZR2022MH211the Key Program of Shandong Provincial Natural Science Foundation,Grant/Award Number:ZR2020KE018National Natural Science Foundation of China,Grant/Award Number:52003068。
文摘Background:Scientific animal models are indispensable for studying trauma repair.This work aimed at establishing a more scientific rat trauma model by studying different rat trauma models caused by different trauma numbers,locations,and trauma attachment tension unloaders and rat age.Methods:A four-trauma self-upper,lower,left and right control model;a two-trauma self-trauma bare and ring control model;and a young and old rat trauma model were created to evaluate the condition of these traumas.Results:In the four-trauma self-control model,the healing status of the upper proximal cephalic trauma was better than that of the lower proximal caudal trauma,whereas there was no significant difference between the left and right trauma.The healing rate and postwound condition of the trauma with a ring control in the two-trauma model were better than those of the bare side.The healing speed of the old rats was slower,and the amount of extracellular matrix in the subcutaneous tissue after healing was significantly lower than that of the young rats.Conclusion:The double trauma with a ring is a more scientific and reasonable experimental model.There is a significant difference between young and old rats in the wound healing process.Therefore,the appropriate age of the rats should be selected according to the main age range of the patients with similar conditions in the clinical setting being mimicked.
基金Supported by 2019 Mongolian Pharmacy"First-Class Discipline"Scientific Research Project of Inner Mongolia Medical University(myxy1xkky2019-17).
文摘[Objectives]To investigate the effects of different doses of chili pepper on inflammatory response factors in rats.[Methods]Sixty male SD rats were randomly divided into blank control group,high-dose chili pepper group,medium-dose chili pepper group,and low-dose chili pepper group.Chili pepper aqueous solution was administered by gavage for three months.One day after the last administration,serum and cerebrospinal fluid were collected to measure the concentrations of hs-CRP and IL-6 in serum,and PCT in cerebrospinal fluid.[Results]Compared with the blank control group,the high-dose,medium-dose,and low-dose chili pepper groups showed significant increases in hs-CRP,IL-6,and PCT levels(P<0.05).[Conclusions]Different doses of chili pepper exerted varying effects on inflammatory factors in rats.
基金Foundation items:Wuhan Health Research Fund(Grant No.WZ21A06)National Natural Science Foundation of China(Grant No.81903815)。
文摘This study aimed to assess the therapeutic potential of Anemarrhenae Rhizoma(AR)in osteoporotic rats and to elucidate the metabolic pathways involved in AR’s role in alleviating osteoporosis(OP).OP was induced in rats through ovariectomy(OVX),followed by oral administration of either high or low doses of AR,as well as estradiol valerate,over a 14-week period.Micro-computed tomography(Micro-CT)was employed to examine the femur tissue morphology,while enzyme-linked immunosorbent assay(ELISA)was adopted to measure serum levels of PINP and CTX-I to evaluate AR’s efficacy in treating OP.Additionally,metabolomic profiling of femur tissues was conducted using gas chromatography-mass spectrometry(GC-MS).The bioactive components of AR,along with its therapeutic targets for OP,were identified through UPLC-MS/MS and online database searches,and metabolic networks were established by integrating differential metabolites and potential targets.Furthermore,Western blotting analysis confirmed key molecular targets.The findings revealed that AR treatment significantly mitigated OVX-induced OP in rats.Metabolomic analysis indicated that AR exerted its effects by modulating the levels of 10 key metabolites(such as linoleic acid and inositol)and influencing five crucial metabolic pathways,including linoleic acid metabolism and the phosphoinositide signaling system.Among these,the linoleic acid metabolic pathway emerged as a pivotal focus for further investigation based on the constructed interaction network of differential metabolites and targets.Western blotting analysis demonstrated that AR reversed the up-regulation of CYP1A2 and CYP2C9,two targets associated with the linoleic acid metabolic pathwa y,in OP rats.In conclusion,AR appeared to ameliorate OP by modulating metabolite levels in OVX rats,with its mechanism of action likely centered on regulating the linoleic acid metabolic pathway.
基金College of Medicine and Public Health,MD,Advanced Studies ProgramAustralian National Health and Medical Research Council(NHMRC),Grant/Award Number:GNT2003683。
文摘The COVID-19 pandemic posed a challenge for clinical management of a new lung disease that was characterized by inflammation,endothelial cell dysfunction,and thrombosis,which occur after the replication phase of infection of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).There are many laboratory models of active SARS-CoV-2 infection in mice,reflecting an acute lung injury in an otherwise healthy animal,but there is a lack of accurate animal models of the postviral inflammatory phase of the COVID-19 lung reflecting severe disease.The monocrotaline(MCT)-treated rat is a widely used laboratory model of pulmonary hypertension(PH).Not often discussed,however,are the observed changes in inflammation,edema,fibrosis,and microthrombosis in the lung prior to PH.At the cellular level,there is loss of pneumocytes and endotheliopathy,and at the molecular level the MCT rat lung is characterized by a proinflammatory cytokine profile,namely elevated interleukin 6,transforming growth factorβand tumor necrosis factor,M1 macrophage phenotype,and dysregulation of the angiotensin converting enzyme(ACE)/ACE2 balance.The systems-level pathophysiology of the MCT-treated rat includes progressive cardiopulmonary dysfunction.The MCT-treated rat clearly differs from the COVID-19 lung in terms of the triggers for pathology,but there are many parallels apparent in both the MCT-treated rat and the COVID-19 lung.The MCT-treated rat lung as a model of the COVID-19 lung may provide an in-depth understanding of the factors that drive the lung to more severe pathology,treatments that benefit lung recovery,or the factors that prove a useful research platform for future emerging respiratory threats of similar pathology.
基金The present work was supported by the National Natural Science Foundation(31970508)the National Key Research and Development Program of China(2022YFF0710702).
文摘Background:C1QL3 is widely expressed in the brain and is specifically produced by a subset of excitatory neurons.However,its function is still not clear.We established C1ql3-deficient rats to investigate the role of C1QL3 in the brain.Methods:C1ql3 knockout(KO)rats were generated using CRISPR/Cas9.C1ql3 KO was determined by polymerase chain reaction(PCR),DNA sequencing,and western blot-ting.Microglia morphology and cytokine expression with or without lipopolysaccha-ride(LPS)stimulus were analyzed using immunohistochemistry and real-time PCR.The brain structure changes in KO rats were examined using magnetic resonance imaging.Neuronal architecture alteration was analyzed by performing Golgi staining.Behavior was evaluated using the open field test,Morris water maze test,and Y maze test.Results:C1ql3 KO significantly increased the number of ramified microglia and decreased the number of hypertrophic microglia,whereas C1ql3 KO did not in-fluence the expression of pro-inflammatory factors and anti-inflammatory factors except IL-10.C1ql3 KO brains had more amoeboid microglia types and higher Arg-1 expression compared with the WT rats after LPS stimulation.The brain weights and HPC sizes of C1ql3 KO rats did not differ from WT rats.C1ql3 KO damaged neuronal integrity including neuron dendritic arbors and spine density.C1ql3 KO rats demonstrated an increase in spontaneous activity and an impairment in short working memory.Conclusions:C1ql3 KO not only interrupts the neuronal integrity but also affects the microglial activation,resulting in hyperactive behavior and impaired short memory in rats,which highlights the role of C1QL3 in the regulation of structure and function of both neuronal and microglial cells.
基金Research Project of China Baoyuan Investment Co.,Ltd,Grant/Award Number:Program CBYI202102Haihe Laboratory of Cell Ecosystem Innovation Fund,Grant/Award Number:HH24KYZX0007+4 种基金CAMS Innovation Fund for Medical Sciences,Grant/Award Number:2021-I2M-1-024,2021-I2M-1-034 and 2023-I2M-2-001Fundamental Research Funds for the Central Universities,Grant/Award Number:3332022040 and 3332023164Open Research Project in State Key Laboratory of Vascular Homeostasis and Remodeling,Peking University,Grant/Award Number:202411State Key Laboratory Special Fund,Grant/Award Number:2060204the Non-profit Central Research Institute Fund of the Chinese Academy of Medical Sciences,Grant/Award Number:2023-PT180-01.
文摘Background:The Cre/loxP system is most popular in mice,but its application in rats has largely lagged far behind.The rat is vital laboratory animal,especially in toxicological and neurological studies.Generating genetic tools to manipulate neurons in rats could benefit neurological research.Methods:Using the CRISPR/Cas9 system,we inserted a Cre cassette into endogenous Thy1 and NeuN loci.Thy1-Cre rats featured a downstream P2A-linked insertion,while NeuN-Cre was inserted at the transcriptional start site.The Cre activity was assessed by crossing with a Cre reporter(Rosa26 imCherry)rat and through analyzing mCherry expression patterns.The specificity of cell type was further confirmed by immunofluorescence with NeuN antibody.Phenotypic consequences were assessed by crossing with ND1^(LSL) rats to deplete ND1,followed by monitoring weight/survival and conducting motor function tests.Results:We generated two neuron-specific rats(Thy1-Cre and NeuN-Cre),which exhibited high neuron-specific Cre expression in brain and spinal cord with minor leakage in other tissues.Thy1-Cre showed minor leakage in spleen,lung and kidney while NeuN-Cre showed minor leakage in spleen and kidney.ND1^(Thy1-Cre) and ND1^(NeuN-Cre) rats both showed decreased body weights and survival times.The ND1^(NeuN-Cre) rats died within two weeks,while ND1^(Thy1-Cre) rats lived longer with impaired motor function.Conclusions:We successfully generated two neuron-specific NeuN-Cre and Thy1-Cre rats,and systemically analyzed their expression pattern.
基金supported by the National Natural Science Foundation of China(32060199,32360197,31971035,and 31771182)the Jiangxi Province Natural Science Foundation(20224ACB206016).
文摘Humans and animals have a fundamental ability to use experiences and environmental information to organize behavior.It often happens that humans and animals make decisions and prepare actions under uncertain situations.Uncertainty would significantly affect the state of animals’minds,but may not be reflected in behavior.How to“read animals’mind state”under different situations is a challenge.Here,we report that neuronal activity in the medial prefrontal cortex(mPFC)of rats can reflect the environmental uncertainty when the task situation changes from certain to uncertain.Rats were trained to perform behavioral tasks under certain and uncertain situations.Under certain situations,rats were required to simply repeat two nose-poking actions that each triggered short auditory tone feedback(single-task situation).Whereas under the uncertain situation,the feedback could randomly be either the previous tone or a short musical rhythm.No additional action was required upon the music feedback,and the same secondary nose-poking action was required upon the tone feedback(dual-task situation);therefore,the coming task was uncertain before action initiation.We recorded single-unit activity from the mPFC when the rats were performing the tasks.We found that in the dual task,when uncertainty was introduced,many mPFC neurons were actively engaged in dealing with the uncertainty before the task initiation,suggesting that the rats could be aware of the task situation change and encode the information in the mPFC before the action of task initiation.
基金CAMS Innovation Fund for Medical Sciences(CIFMS),Grant/Award Number:2021-I2M-1-035National Natural Science Foundation of China,Grant/Award Number:31970508。
文摘Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed rat resource has rapidly increased,but the liver-specific Cre rat lines for studying liver diseases and interested genes are limited,especially in a spatially and temporally restricted manner.Methods:RNA sequencing and real-time polymerase chain reaction(PCR)were used to screen and confirm the presence of liver-specific genes.Apoa4-Cre rats and Cyp2c11-Cre rats were produced by CRISPR/Cas9 knockin.Rosa26-imCherry rats were employed to hybridize with the Cre rats to obtain the Apoa4-Cre/Rosa26-imCherry and Cyp2c11-Cre/Rosa26-imCherry rats.The temporal and spatial patterns of Cre expression were determined by the observation of red fluorescence on tis-sue sections.Hematoxylin-eosin stain was used to evaluate the liver histopathologic changes.The blood biochemical analysis of several liver enzymes and liver lipid profile was performed to evaluate the liver function of Cre rats.Results:Apoa4 and Cyp2c11 were identified as two liver-specific genes.Apoa4-Cre and Cyp2c11-Cre rats were produced and hybridized with Rosa26-imCherry rats.The red fluorescence indicated that the Cre recombinases were specially expressed in the juvenile and adult liver and not in other organs of two hybridized rats.All the blood biochemical parameters except low-density lipoprotein(LDL)did not change signifi-cantly in the Cre rats.No histological alterations were detected in the livers of the Cre rats.Conclusions:Liver-specific Apoa4-Cre and Cyp2c11-Cre rats have been established successfully and could be used to study gene knockout,specifically in juvenile and adult liver.
文摘Although Steen's solution plays a crucial role in maintaining vascular oncotic pressure and reducing pulmonary edema due to its high concentration of human serum albumin during ex vivo lung perfusion(EVLP),it incurs high costs and potential immune responses in rat models.In our preliminary research,donor lungs perfused with Steen's solution exhibited significant lung injury post transplantation,likely due to a strong immune response elicited by the high concentration of xenogenic proteins upon reperfusion.This study explores a low-cost and efficient perfusate composition for use in rat EVLP and lung transplant models.The modified perfusate employed in this study uses artificial colloids as the primary component,with an albumin concentration of only 2%,demonstrating comparable physiological parameters,metabolic levels,and pathological changes to those of Steen's solution during EVLP.Moreover,the modified composition exhibited good biocompatibility in transplantation,with minimal lung injury.In summary,the proposed perfusate offers a safer and more economical alternative for establishing rat EVLP and lung transplant models,facilitating the advancement of related research.
