Comparative Study of Staphylococcus epidermidis Virulence Profiles in an animal Model of Endophthalmitis

Introduction: Gram-positive staphylococci are responsible for over 90% of cases of postoperative infectious endophthalmitis, with Staphylococcus epidermidis accounting for approximately 70% of isolated microorganisms. Objective: To evaluate the relationship between S. epidermidis virulence and severity of endophthalmitis in an animal model. Methodology: New Zealand albino rabbits were divided into two groups and administered 100 colony-forming units of S. epidermidis strains. In the virulent group, four rabbits received a multi-antibiotic resistant, mecA, ica, and atlE gene carrying S. epidermidis strain, from a patient´s conjuntival microbiota and three a biofilm-forming S. epidermidis ATCC 35984. In the non-virulent group, five rabbits were inoculated with a strain sensitive to all tested antibiotics and lacking mecA, ica, and atlE genes, also from a patient and three rabbits received the non-producer biofilm S. epidermidis ATCC 29122. Clinical and ultrasound examinations were conducted every three hours until endophthalmitis symptoms appeared, followed by daily clinical assessments. Histological evaluations were performed 15 days post-inoculation. Results: The less virulent group displayed milder inflammation and reduced intraocular damage in comparison to the more virulent group based on clinical and ultrasound observations. Nevertheless, histopathological analysis revealed similar inflammation in all groups, 15 days post-inoculation. Discussion: Less virulent S. epidermidis strains induced less severe inflammation as observed through clinical and ultrasound assessments. However, long-term histopathological assessments showed effects comparable to those seen with the more virulent strain.

Introducción: Gram-positive staphylococci are responsible for over 90% of cases of postoperative infectious endophthalmitis, with Staphylococcus epidermidis accounting for approximately 70% of isolated microorganisms. Objetivo: Evualuar la relación entre la virulencia de S. epidermidis y la gravedad de la endoftalmitis en un modelo animal. Metodología: Se dividieron conejos albinos de Nueva Zelanda en dos grupos y se les administraron 100 unidades formadoras de colonias de cepas de S. epidermidis. En el grupo virulento, cuatro conejos recibieron una cepa multirresistente de S. epidermidis portadora de gen mecA, ica y atlE, de la microbiota conjuntival de un paciente y tres conejos la cepa S. epidermidis ATCC 35984 formadora de biopelículas. En el grupo no-virulento, se inocularon cinco conejos con una cepa sensible a todos los antibióticos probados y que carecía de los genes mecA, ica y atlE, también de un paciente y tres conejos recibieron la cepa S. epidermidis ATCC 29122 no productora de biopelícula. Se realizaron exámenes clínicos y ecográficos cada tres horas hasta la aparición de síntomas de endoftalmitis, seguido de evaluaciones clínicas diarias. Las evaluaciones histológicas se realizaron 15 días después de la inoculación. Resultados: El grupo menos virulento mostró una inflamación más leve y un daño intraocular reducido en comparación con el grupo más virulento según observaciones clínicas y ecográficas. Sin embargo, el análisis histopatológico reveló una inflamación similar en todos los grupos, 15 días después de la inoculación. Discusión: Las cepas de S. epidermidis menos virulentas indujeron una inflamación menos grave, como se observó mediante evaluaciones clínicas y ecográficas. Sin embargo, las evaluaciones histopatológicas a largo plazo mostraron efectos comparables a los observados con la cepa más virulenta.

Fitting Degrees of Animal Models of Diarrhea-irritable Bowel Syndrome with Clinical Characteristics of Western Medicine and Traditional Chinese Medicine / 中国实验方剂学杂志

Diarrhea-irritable bowel syndrome （IBS-D） is one of the common functional bowel diseases in clinical practice. Since it pathogenesis is complex and has not been fully elucidated， effective treatment methods remains to be developed for this disease. Establishing the animal models of IBS-D in accordance with the clinical characteristics of traditional Chinese medicine （TCM） and Western medicine helps to reveal the pathogenesis of this disease and improve the treatment plan. The fitting degree of an animal model with clinical characteristics is an indicator to evaluate the effectiveness of the animal model in simulating the disease characteristics of Western medicine and the syndromes of TCM based on the latest diagnostic standards. By reviewing the relevant articles about the animal models of IBS-D， we discovered that rats were the preferred animals for modeling， and the models were mainly induced by single factors， double factors， or the combination of multiple factors. The established animal models mainly present symptoms or signs associated with visceral hypersensitivity or/and gastrointestinal motility abnormalities. The single factor-induced rat models of IBS-D had high fitting degrees with the clinical characteristics of Western medicine but low fitting degrees with the TCM syndromes. The animal models induced by two or more factors had high but varied fitting degrees with the clinical characteristics of Western medicine. In addition， the animal models of IBS-D considering TCM syndromes mainly focuses on the syndrome of liver depression and spleen deficiency， and few models were established for the syndromes of spleen-kidney Yang deficiency， spleen-stomach dampness-heat， spleen deficiency and dampness excess， and cold and heat in complexity. Therefore， it is essential to improve the existing or develop new animal models of IBS-D in the future， so as to provide more tools for deciphering the mechanisms of TCM and Western medicine and developing treatment methods for this disease.

Effect of glutaminase 1 inhibitor on carbon tetrachloride-induced liver fibrosis in mice / 中国生物制品学杂志

@#Objective To investigate the effect of glutaminase 1(GLS1)specific inhibitor BPTES[bis-2-（5-phenylacetamido-1,3,4-thiadiazol-2-yl）ethyl sulfide]on the liver fibrosis in the mouse model of liver fibrosis induced by carbon tetrachloride(CCl4).Methods Male C57BL/6J mice were intraperitoneally injected with olive oil(control group),10%CCl4(10 μL/g,model group)or 10% CCl4(10 μL/g)+ BPTES(10 mg/kg,treatment group),with 10 mice in each group,two doses a week for four weeks to establish liver fibrosis model. Collagen deposition in mouse liver tissue was observed by Sirius red staining. The expression levels of actin alpha 2(Acta2),collagen typeⅠalpha 1(Col1a1)GLS1 and GLS1 protein were detected by qRT-PCR and immunohistochemical staining.Results Compared with the control group,the liver tissue of mice in the model group was generally enlarged,the surface was not smooth and granular,and the ratio of liver mass to tibia length significantly increased(t = 2. 979,P < 0. 05);The Sirius red positive area of collagen deposition increased signifi-cantly in the liver tissue of mice in the model group(t = 7. 661,P < 0. 01),the relative expression levels of Acta2 and Col1a1 significantly increased(t = 4. 335 and 5. 319,respectively,each P < 0. 01),and the mRNA and protein levels of GLS1 significantly increased(t = 5. 319 and 9. 725,respectively,each P < 0. 01). However,compared with the model group,the BPTES treatment group had a reduction in liver mass,a significant reduction in the Sirius red positive area of collagen deposition in liver tissue(t = 7. 427,P < 0. 01),and a significant reduction in the relative expressions of Atca2 and Col1a1(t = 3. 713 and 2. 628,respectively,each P < 0. 05).Conclusion Inhibition of GLS1activity can significantly improve the degree of liver fibrosis induced by CCl4,providing a new idea for the treatment of liver fibrosis.

Animal Models of Ovarian Cancer Based on Characteristics of Clinical Symptoms of Traditional Chinese and Western Medicine / 中国实验方剂学杂志

ObjectiveOvarian cancer is the third most common gynecologic cancer worldwide， with the second highest mortality rate among gynecologic cancers， and age-standardized rates are gradually increasing in many low- and middle-income countries. At present， its etiology and pathogenesis are not clear. There are no obvious symptoms in the early stage， and when the symptoms become obvious， it often indicates the advanced stage. The 5-year survival rate of the advanced stage is only 17%， which poses a great threat to women's health. Therefore， an in-depth study of the etiology and pathogenesis of ovarian cancer is very important to the exploration of prevention and treatment methods for ovarian cancer. Based on the clinical characteristics of ovarian cancer in traditional Chinese and Western medicine， and combined with the existing evaluation methods of animal models， this study evaluated the animal model of ovarian cancer， and provided analysis and suggestions. MethodThis study searched China National Knowledge Infrastructure （CNKI）， Wanfang data， VIP information database， and PubMed database using the keywords "ovarian cancer" and "animal model"， excluded the articles that did not meet the criteria， and then classified the remaining studies. Combined with the clinical diagnostic criteria of Western medicine and traditional Chinese medicine syndrome differentiation， the related indicators of ovarian cancer animal models were assigned and the degree of agreement was evaluated. ResultThe use of the transplanted animal model exhibited the highest frequency， followed by that of the induced model. The degree of agreement of traditional Chinese medicine for each model was lower than that of Western medicine. The induced ovarian cancer model had a high degree of clinical agreement and was similar to human ovarian cancer in terms of tumor growth pattern， disease progression and complications， which is an ideal animal model of ovarian cancer. Although this animal model can simulate the etiology and pathogenesis of ovarian cancer to a certain extent and reflect some indicators of traditional Chinese and Western medicine， it lacks differentiation of traditional Chinese medicine syndromes. ConclusionOn the basis of the original model， the animal model of ovarian cancer was added with Qi deficiency syndrome， blood deficiency syndrome， Qi stagnation syndrome， blood stasis syndrome， heat-toxin syndrome， and Yang deficiency syndrome to establish an animal model combining traditional Chinese medicine disease and syndrome of ovarian cancer， which could better simulate the clinical actual situation of traditional Chinese and Western medicine and lay a solid foundation for the study of integrated traditional Chinese and Western medicine for the treatment of ovarian cancer.

Application of Animal Models of Corneal Neovascularization Based on Data Mining / 中国实验方剂学杂志

ObjectiveTo provide a reference for the establishment of an ideal corneal neovascularization （CNV） animal model by summarizing the modeling characteristics of CNV animal models. MethodWith "CVN" as the theme word， this paper searched the China National Knowledge Infrastructure （CNKI）， Wanfang， Chinese medical journals full-text database， and PubMed database and screened out relevant literature on CNV animal experiments from 2013 to 2023. The database was established by Excel 2021， and the experimental animal strain， gender， modeling method， detection index， and application category were sorted out. The characteristics of the CNV animal model were analyzed. ResultAfter comparative analysis， it was found that the animal strains were Sprague-Dawley rats （87 times， 29.49%） and New Zealand white rabbits （52 times， 17.63%）. Male animals were recommended. Most modeling methods for efficacy verification and mechanism studies were the alkali burn method. Index detection methods included apparent index observation， histopathological detection， immunohistochemistry （IHC）， Western blot， and various polymerase chain reaction （PCR） tests. Detection indexes included apparent indication， corneal histopathology， CNV regulation， etc. ConclusionThe CNV model of SD rats induced by the alkali burn method is recommended for model replication， and the indexes are mainly selected from the growth of CNV， corneal histopathological test， and vascular endothelial growth factor （VEGF）-related test. In addition， according to the demand， the corneal apparent indication and the basic indexes related to the regulation of CNV， such as vascular endothelial growth factor receptor 2 （VEGFR2）， basic fibroblast growth factor （bFGF）， and secretogranin Ⅲ （Scg3） are also selected. Clinical treatment of CNV relies on anti-inflammatory drugs and anti-VEGF drugs， and there is a lack of application of traditional Chinese medicine （TCM）， so the model needs to be improved by adding elements of TCM syndromes.

Establishment and Evaluation of Animal Model of Acute Myocardial Infarction with Syndrome of Qi and Yin Deficiency / 中国实验方剂学杂志

ObjectiveTo explore the establishment and evaluation methods of the rat model of acute myocardial infarction （AMI） in coronary heart disease with the syndrome of Qi and Yin deficiency by sleep deprivation （SD） combined with isoproterenol （ISO） and preliminarily explore its biological basis. MethodForty SD rats were assigned into normal （no treatment）， SD （treatment in modified multi-platform water environment for 96 h）， ISO （subcutaneous injection of ISO at 100 mg·kg-1 once every other day for a total of 2 times）， and SD+ISO （injection of 100 mg·kg-1 ISO after SD for 72 h and 96 h） groups. The cardiac function was detected by small animal echocardiography. The serum levels of creatine kinase （CK）， creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， and cardiac troponin T （cTnT） were measured by biochemical methods. The pathological changes of the myocardial tissue were observed by hematoxylin-eosin staining. The general state， body weight， grip strength， body temperature， behaviors in open field test， serum levels of cyclic adenosine monophosphate （cAMP）， cyclic guanosine monophosphate （cGMP）， cAMP/cGMP ratio， red （R）， green （G）， blue （B） values of the tongue surface， and pulse amplitude were observed and measured to evaluate the modeling results. Enzyme-linked immunosorbent assay was employed to determine the serum levels of interleukin-18 （IL-18）， tumor necrosis factor-α （TNF-α）， superoxide dismutase （SOD）， malondialdehyde （MDA）， corticotropin-releasing factor （CRF）， adrenocorticotropic hormone （ACTH）， triiodothyronine （T3）， tetraiodothyronine （T4）， cluster of differentiation 4 （CD4）， and cluster of differentiation 8 （CD8）. ResultIn terms of disease indicators， the ISO and SD+ISO groups had lower cardiac function indicators than the normal group （P<0.01）. The levels of CK， CM-MB， LDH and cTnT elevated in each model group compared with the normal group （P<0.01）. The pathological changes of myocardial tissue were obvious in the ISO and SD+ISO groups. In terms of syndrome indicators， compared with the normal group， the SD and SD+ISO groups showed decreased body weight at each time point （P<0.01）， and the ISO group showed decreased body weight at the time points of 48 h and 72 h （P<0.05， P<0.01）. The paw temperature and rectal temperature increased in the SD group （P<0.01）. The model groups showed weakened grasp strength， lowered R， G， and B values of the tongue surface （P<0.01）， prolonged immobility time （P<0.01）， reduced total distance and number of entering the central area （P<0.01）， decreased average speed （P<0.05， P<0.01）， and increased cAMP and cGMP （P<0.05， P<0.01）. The cAMP/cGMP ratio was increased in the SD+ISO group （P<0.01）， and the pulse amplitude was decreased in the SD and SD+ISO groups （P<0.01）. In terms of serological indicators，compared with the normal group， the levels of IL-18， TNF-α， SOD and MDA were significantly increased in the ISO and SD+ISO groups （P<0.01）， the CRF， ACTH， CORT， T3， T4， CD4 and CD8 in the model groups were increased （P<0.05， P<0.01）. ConclusionSleep deprivation for 96 h combined with high-dose ISO can successfully establish a rat model of acute myocardial infarction in coronary heart disease with the syndrome of Qi and Yin deficiency. The model evaluation system can be built with disease indicators of western medicine， histopathological indicators， macroscopic indicators of traditional Chinese medicine， and serological indicators.

Research progress on animal models of combined radiation-wound injury / 中国辐射卫生

A combined radiation-wound injury refers to a radiation injury combined with a traumatic wound, with the characteristics of repeated ulceration and a long and difficult healing process, which is a focus in the field of research on difficult-to-heal wounds. To research combined radiation-wound injuries, the establishment of animal models is a key part, and appropriate animal models are a guarantee of reliable experimental results. This review summarizes the current research progress on various animal models of combined radiation-wound injuries in terms of radiation types, animal species, and injury types and location, aiming to provide a scientific basis for establishing standardized animal models, studying injury mechanisms, and evaluating prevention and treatment efficacy for combined radiation-wound injuries.

Establishment and application of animal models of myocardial infarction / 解剖学报

Myocardial infarction is one of the severe cardiovascular diseases.The patients with myocardial infarction die of heart failure or arrhythmia.In recent years,the studies in myocardial infarction therapies have advanced greatly,especially the preclinical experimental studies.The experimental studies of myocardial infarction often rely on animal models.Therefore,successful establishment of the myocardial infarction models has important application value in exploring the new techniques and measures for repairing the infarcted myocardium.In this paper,the techniques in establishment of the myocardial infarction models and strategies of their application are summarized.

The application of low temperature-storing tumor lumps in vitro in establishing VX2 liver cancer models:study of its modeling effect / 介入放射学杂志

Objective To compare the effect of using the fresh tumor lump and the recovered low temperature-storing VX2 tumor lump that have been frozen for different time to construct the rabbit HCC model.Methods Fish-like fresh VX2 tumor lumps were selected.After the peripheral necrotic tissue and muscle were removed,the tumor lumps were frozen at-80℃ for 3,5 and 7 months.Twenty New Zealand white rabbits were randomly divided into 4 groups.The rabbits in group A(control group)received fresh liver tumor lump implantation to construct in-situ VX2 rabbit HCC models.The rabbits in groups B,C and D(experimental groups)received implantation of the recovered low temperature-storing VX2 tumor lump which had been frozen at-80℃ for 3,5 and 7 months,respectively,to construct in-situ VX2 rabbit HCC models.Fourteen days after implantation,the modeling effect of tumor formation in each group was assessed.The proliferation,apoptosis of tumor cells and the angiogenesis were evaluated by immunofluorescence assay.Results The tumor formation rate of all group A,B,C and D was 100％.However,with the extension of cryopreservation time,the difference in tumor mass activity became larger after 5 months,and the necrotic area of liver tumor center became enlarged.Histological examination showed that there were no significant differences in the expressions of TUNEL,Ki67,HIF-α,VEGF and CD31 between group A and group B,while there were significant differences in the expressions of TUNEL,Ki67,HIF1-α,VEGF and CD31 between group A,B and group C,D.Conclusion The rabbit VX2 HCC model,which is constructed by implantation of recovered low temperature-storing VX2 tumor lump being frozen at-80℃ in vitro for a certain time,can be successfully established within 7 months.The difference in tumor mass activity between tumor lumps became larger after 5 months.But on the whole,the constructed rabbit VX2 HCC model can better preserve the tumor strain activity.This modeling technique can save manpower and material resources.(J Intervent Radiol,2024,33:269-274)

Two golden hamster models of hypervirulent Klebsiella pneumoniae respiratory infection:a comparative study / 军事医学

Objective To establish two golden hamster models infected with hypervirulent Klebsiella pneumoniae via aerosolized intratracheal(i.t.)and intranasal(i.n.)inoculation,and compare their properties.Methods Golden hamsters of 4 to 5 weeks old were exposed to K.pneumoniae NTUH-K2044 via i.t.route and i.n.route respectively.The survival of these golden hamsters was observed and recorded within 14 days of infection before the 50%lethal dose(LD50),survival rate,bacterial respiratory deposition rate,lung bacterial load and histopathology of the infected golden hamsters in the two groups were detected.Results The LD50 of the i.t.route(3×104 CFU)was lower than that of the i.n.route(7×105 CFU)in golden hamsters.After 4×106 CFU NTUH-K2044 infection,the golden hamsters in the i.t.group had 96.46%of the bacteria deposited and colonized in the lung,developed lobar pneumonia and died without exception within 4 days of infection,while those in the i.n.group had 95.62%of the bacteria deposited in the mouth and nose initially before the bacteria moved down to the trachea for colonization and were cleared out gradually.This group mainly acquired bronchopneumonia with relatively mild lung lesions,with a 14-day survival rate of 70%.Conclusion Inoculation routes can make a difference to the disease type of respiratory tract infections in animal models.The i.t.route mainly causes lobar pneumonia with severe lung lesions,while the i.n.route leads to bronchopneumonia with mild lung lesions.The two animal models established above may be utilized for pathogenesis investigation and treatment efficacy evaluation of Klebsiella pneumoniae.

Construction of the pore-forming toxin gene exlA knock-out mutant of Pseudomonas aeruginosa and its basic characteristics / 军事医学

Objective To construct a non-trace deletion mutant of exlA in Pseudomonas aeruginosa strain NY8755(NY8755ΔexlA)and investigate the basic characteristics of pore-forming toxin ExlA.Methods The NY8755ΔexlA was constructed using the secondary homologous recombination method.C57BL/6J female mice ages 6 to 8 weeks were infected with NY8755 and NY8755 ΔexlA via aerosolized intratraheal inoculation respectively.Within 7 days of infection,the survival and weight changes of the mice were observed and recorded before the proinflammatory cytokines in the bronchoal-veolar lavage fluid(BALF)of the infected mice in the two groups were detected.Results The sequencing results showed that NY8755 ΔexlA was constructed.After 1×107 CFU NY8755 and NY8755 ΔexlA were infected,all the mice in the wild-type strain group died within 48 hours,while those in the mutant strain group began to die after 48 hours,and 40%of them remained alive 7 days later.The weight of surviving mice in the mutant strain group decreased but recovered gradually.After 12 hours of infection,there were more bloody exudates(redder in color)in the BALF of the wild-type strain group than in the mutant strain group,and the contents of proinflammatory cytokines interleukin-1β(IL-1β)and interleukin-17A (IL-17A)were significantly different. Conclusion Pseudomonas aeruginosa pore-forming toxin ExlA is the key pathogenic virulence factor of the exlA-positive Pseudomonas aeruginosa,which can significantly affect the survival status of mice and cause obvious inflammation in mice. Very little information is available on the action mechanisms of ExlA. In this study, The NY8755ΔexlA and the C57BL/6J mouse models infected with NY8755 and NY8755ΔexlA have been constructed that may be used for the investigation of pathogenesis of exlA-positive Pseudomonas aeruginosa.

Research advances in animal models of tricuspid regurgitation / 复旦学报（医学版）

Tricuspid regurgitation(TR)cases are widely distributed in China.Poor clinical drug efficacy,high surgical risk,and poor prognosis for right heart failure are found in patients with moderate or severe TR.In recent years,with the innovation of valve instruments and the development of technology,transcatheter tricuspid valve treatment could be a new choice for high-risk TR patients in surgery.Many TR animal models have emerged these years for the research of the mechanism of TR and for the clinical verification of instruments.Therefore,this review focuses on how to develop an animal model of TR and discusses the advantages and disadvantages of these techniques.

Research progress in animal models for tic disorder / 中华实用儿科临床杂志

Tic disorder (TD) is a neurodevelopmental disorder that occurs in childhood, characterized by involuntary movements or vocal tics and occasionally accompanied by various comorbidities.It may seriously affect children′s daily life, learning and social activities.Exploring its etiology and pathological mechanisms and developing drugs require extensive research on animal models.There are currently various methods for constructing TD models, including induced and spontaneous animal models, such as induced animal models with abnormal neurotransmitters and immune functions and spontaneous animal models based on genetic engineering.However, no animal model can fully match the manifestations of TD.This article summarizes the commonly used animal models for TD to provide references for exploring pathological mechanisms and developing drugs.

Establishment of animal model of cervicofacial venous malformations with external jugular vein in sheep / 实用口腔医学杂志

Objective:To establish the animal model of cervicofacial venous malformations(VMs)by surgical reconstruction of exter-nal jugular vein in sheep.Methods:The external jugular veins of 5 sheep were dissected,and the position,course,branch and exter-nal diameter were observed and measured.The models of VMs with draining and returning veins were constructed by suturing or constric-ting the proximal part of main trunk and ligating or constricting the distal part of the jugular or branch veins.The animal model was eval-uated by Doppler ultrasound,gross observation and histological observation at the 4th week after surgery.Results:The external jugular veins of sheep is in the lateral side of bilateral neck,and the main trunk is formed by the maxillary vein and lingual facial vein.The ex-ternal diameter ranges from 6 to 12 mm,with an average external diameter of 9.3 mm.Immediately after the external jugular vein was sutured and narrowed at the proximal part of the main vein,the distal part of the vein branch was ligated or narrowed,the blood flow speed slowed down and the veins in the model area bulged.4 weeks after surgery,gross observation showed that most veins narrowed and thrombosis was formed in part of the venous lumen.The central region of some specimens was dilated,and the peripheral collateral veins were dilated in some models.Doppler ultrasonography showed that the lumens of most veins were dilated and the returning veins and the inflow veins were narrowed.Colored blood flow was seen in the lumen.Histological observation showed that the structure of vein endothelium and wall was close to the normal vein,and the vein vessel wall of some specimens was thickened.Conclusion:The VMs model estab-lished by external jugular vein of sheep basically meets the re-quirements and is expected to be used in the therapeutic meth-odology research of cervicofacial VMs.

A study on the animal model preparation and early histological changes of trans-sutural distraction osteo-genesis / 实用口腔医学杂志

Objective:To establish an animal model of trans-sutural distraction osteogenesis in SD rats.Methods:A self-designed V-shaped distraction device(distractor)was fabricated with the traction force(N)of 0,1.3,2.2,3.0,4.3 and 5.0 corresponding to the distraction length(mm)of 5,4,3,2,1 and 0 respectively,meeting the trans-sutural distraction osteogenesis requirements in skull of 5-week-old SD rats.The distractor was plased into the sagittal suture of 12 SD rats.Continuous sampling was conducted 1,3,5 and 7 days respectively(n=3)after operation.The tissue changes in the trans-sutural distraction area were observed by HE and Masson's trichrome staining.Inflammation levels were determined using Arg-1 immunofluorescence staining.The early angiogenesis was clarified through co-staining with CD31 and EMCN.Results:A stable trans-sutural distraction osteogenesis model was estab-lished,5 mm distraction osteogenesis width was observed completely within 7 days of distraction.Significant new bone formation was observed at 7 days after operation.Arg-1 expression increased and was concentrated at the bone margins,overlapping with the areas of new bone formation.EMCN expression gradually decreased,and by day 7 CD31 was predominant,indicating the basic maturation of blood vessels.Conclusion:This study successfully constructed a stable and effective trans-sutural distraction osteogenesis animal model,and provides an experimental basis for the investigation of its early continuous histological changes.

Molecular docking analysis of the anti-inflammatory mechanism of Cibotium barometz and Epimedium for rheumatoid arthritis:animal experiment validation / 中国组织工程研究

BACKGROUND:In clinical practice,Cibotium barometz and Epimedium have shown significant efficacy in the treatment of rheumatoid arthritis,but the complex active ingredients contained in the two have an unclear mechanism of action at the molecular level for the treatment of rheumatoid arthritis. OBJECTIVE:Based on network pharmacology and molecular docking technology,to establish a collagen-induced arthritis model and to verify the potential targets and pathways of Cibotium barometz and Epimedium in the treatment of rheumatoid arthritis,providing reliable experimental evidence for the use of clinical formulas with Cibotium barometz and Epimedium as the main components. METHODS:Utilizing traditional Chinese medicine research platforms,traditional Chinese medicine encyclopedias,and databases of traditional Chinese medicine and chemical components from the Shanghai Institute of Organic,effective ingredients were retrieved and identified.3D molecular formulas were obtained from the PubChem platform and target predictions were made using PharmMapper and SwissTargetPrediction.Disease targets for rheumatoid arthritis were obtained from gene databases such as DrugBank,GeneCards,and OMIM.The intersections of targets and Cibotium barometz and Epimedium were plotted using VENNY 2.1 after calibration with the Uniport database.A protein-protein interaction network graph was constructed using the STRING platform.Gene Ontology function analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were performed using the Metascape platform for data visualization.A four-layered network model of traditional Chinese medicine,ingredients,targets,diseases,and pathways was constructed using Cytoscape 3.9.0.The main effective ingredients were docked with core targets using AutoDock-Vina software to explore the best binding targets.A type II collagen+adjuvant-induced arthritis rat model was established,and the effects of Cibotium barometz and Epimedium on relevant pathway targets and inflammatory cell factors were observed after 21 days of intervention. RESULTS AND CONCLUSION:A total of 28 active ingredients from Cibotium barometz and Epimedium were selected,yielding 288 intersection targets for rheumatoid arthritis.The main ingredients included isobavachalcone,cibotium,and epimedium.The main targets included protein kinase 1 for serine/threonine(AKT1),tumor necrosis factor,and vascular endothelial growth factor A.Gene ontology analysis yielded 2 232 biological processes,mainly related to serine protein phosphorylation,positive regulation of serine/threonine protein kinase,and reactive oxygen metabolism.Kyoto Encyclopedia of Genes and Genomes enrichment analysis yielded 202 pathways,mainly involving the PI3K/AKT signaling pathway and epidermal growth factor receptor signaling pathway,which may exert therapeutic effects by regulating synovial cell apoptosis and proliferation and suppressing inflammatory factors.Molecular docking results showed the strongest binding activity and stable structure of Cibotium barometz and Epimedium with AKT1 and estrogen receptor transcription factor 1,which was closely related to apoptosis and proliferation and inflammatory signaling pathways such as PI3K/AKT.Cibotium barometz and Epimedium reduced the expression of interleukin-1β,interleukin-6,and tumor necrosis factor-α in the serum of collagen-induced arthritis rat models.Cibotium barometz and Epimedium reduced the expression of p-PI3K,p-AKT,and p-FOXO1 in the synovium of collagen-induced arthritis rat models.The results indicate that the combination of Cibotium barometz and Epimedium may exert therapeutic effects by inhibiting the proliferation of synovial cells and suppressing the expression of inflammatory factors via the PI3K/AKT/FOXO1 signaling pathway.This may be closely related to the occurrence of inflammation and bone destruction in rheumatoid arthritis,and provides a reference for the rational use and development of new drugs in clinical practice.

Establishment and evaluation of a streptozotocin-induced diabetic encephalopathy rat model / 中国组织工程研究

BACKGROUND:Animal models of diabetic encephalopathy that have been studied mainly include streptozotocin-induced model,high-sugar and high-fat diet-induced model and spontaneous animal model.Establishing a simple,easy,short-cycle,safe and effective model of diabetic encephalopathy can help to explore the subsequent pathogenesis and screen therapeutic drugs. OBJECTIVE:To further explore and evaluate the method of building diabetic encephalopathy rat models. METHODS:Twenty Sprague-Dawley rats were randomly divided into control(n=10)and model(n=10)groups.Rats in the model group were given a single injection of 45 mg/kg streptozotocin in the left lower abdominal cavity,and those in the control group were given the same amount of citrate buffer.During the experiment,the body mass,feed intake,water intake and blood glucose were measured.After 8 weeks,the glucose tolerance and oxidative stress levels were measured,and the pathological changes of brain tissue and the expression of apoptotic proteins were compared between groups. RESULTS AND CONCLUSION:Compared with the control group,the food intake,water intake,encephalization quotient,blood glucose and area under the blood glucose curve were significantly increased in the model group,while the body mass decreased significantly(P<0.01).Histopathological examination of the brain showed that compared with the control group,the number of surviving nerve cells was significantly reduced in the model group(P<0.01),with more significant pathological damage of nerve cells.Compared with the control group,the activities of serum superoxide dismutase,catalase and glutathione in the model group were significantly decreased(P<0.01),and the content of oxidative malondialdehyde was significantly increased(P<0.05).The expression levels of apoptosis-related proteins Bax and Caspase-3 in brain tissue increased in the model group compared with the control group,while the expression of Bcl-2 decreased(P<0.01).In conclusion,an 8-week injection of 45 mg/kg streptozotocin can cause obvious pathological damage to the brain tissue of diabetic rats,to successfully establish the rat model of diabetic encephalopathy.

Animal models of femoral bone defects:preparation status and characteristics / 中国组织工程研究

BACKGROUND:The repair and clinical outcome of bone defects remains a hot and difficult area of clinical research,which is a common problem that plagues clinicians.Constructing suitable,reproducible and infinitely close to clinical animal experimental models and their scientific evaluation are essential for further clinical treatment of related diseases. OBJECTIVE:To retrospectively analyze the preparation methods and characteristics of common animal models of femoral bone defects and to assess their strengths and weaknesses,thereby providing some reference for relevant researchers to select appropriate animal models of femoral bone defects. METHODS:PubMed,Web of Science,Medline,and CNKI were retrieved for relevant literature published from January 1,2000 to August 1,2022.The keywords were"bone defect,bone,bones,defect,defects,defective,animal model,animal,model,laboratory,laboratory animal,animal laboratory"in English and"bone defect,animal model,experiment"in Chinese. RESULTS AND CONCLUSION:Twenty-seven randomized controlled animal experiments involving rats,mice,New Zealand rabbits,and sheep were included,analyzed and assessed.The most common types of bone defects were cylindrical bone defects and segmental osteotomy bone defects,generally found in the middle and distal femur.These models are mostly used to evaluate the effects of bone repair materials,drugs,drug-loaded active substances and physical therapy on bone defect repair and explore defect healing mechanisms,particularly the weight-bearing bone defect repair mechanism.Different defect kinds and femoral bone defect ranges have been found in different animal experiments.Researchers can select the suitable animal model and bone defect type based on the goal of the experiment and then set an acceptable bone defect value.Current studies have shown that cylindrical and segmental osteotomy-induced bone defects,mainly in the distal and middle femur,are mostly used in the animal models of femoral bone defects and that the surgical methods and postoperative management are more mature and operable to provide mature experimental animal models.In terms of cylindrical bone defects,rats and New Zealand rabbits are more suitable,whereas segmental osteotomy has no special requirements and all types of animals can meet the experimental requirements.

Network meta-analysis of the modeling effects of different factors on rabbit models of steroid-induced osteonecrosis of femoral head / 中国组织工程研究

OBJECTIVE:The rabbit model of steroid-induced osteonecrosis of femoral head is the most commonly used animal model of femoral head necrosis.The pathological changes of the femoral head are close to clinical practice,however,the conditions,methods and evaluation standards of animal models reported in and outside China are not uniform,which leads to the low scientific value of animal models and is difficult to popularize.This study aimed to clarify the influence of different mold-making conditions on the establishment of steroid-induced osteonecrosis of femoral head rabbit model and analyze the appropriate conditions for the successful model establishment. METHODS:We searched the CNKI,WanFang,VIP,CBM,WoS,PubMed and EMbsae databases for the literature on the modeling of steroid-induced osteonecrosis of femoral head rabbits up to April 1,2022,completed the screening of the literature according to the inclusion and exclusion criteria and literature quality evaluation,and extracted the outcome index data in the literature.RevMan Stata and ADDIS statistical software were used to conduct a meta-analysis of the included data. RESULTS:(1)A total of 82 articles with 1 366 rabbits were included in the study.The steroid-induced osteonecrosis of femoral head modeling methods were divided into three types:steroid-alone method,steroid combined lipopolysaccharide method and steroid combined serum method.Among these,33 articles used steroid-alone method;20 articles used steroid combined lipopolysaccharide method;29 articles used steroid combined serum method.(2)Meta-analysis results showed that the three modeling methods significantly increased the rate of empty bone lacunae in the femoral head of steroid-induced osteonecrosis of femoral head rabbits(P<0.001),and significantly decreased the ratio of the trabecular bone area in the femoral head of steroid-induced osteonecrosis of femoral head rabbits(P<0.001).The order of empty bone lacunae rate of each modeling method was:steroid combined with lipopolysaccharide method>steroid-alone method>steroid combined with serum method>normal group,and the order of trabecular bone area rate of each modeling method was:normal group>steroid combined with serum method>steroid-alone method>steroid combined with lipopolysaccharide method.(3)The results of subgroup analysis suggested that the rate of empty bone lacunae in the rabbit model induced by steroid alone might be related to the rabbit variety and the type of steroid used for modeling(difference between groups P<0.05),in which the combined effect amount of New Zealand white rabbits was higher than that of Chinese white rabbits(P<0.05)and Japanese white rabbits,and the combined effect amount of dexamethasone was higher than that of other steroids.The rate of empty bone lacunae induced by steroid combined with lipopolysaccharide was related to the administration mode of lipopolysaccharide and the type of steroid(P<0.05),among which the combined effect of methylprednisolone sodium succinate was significantly higher than that of other steroids(P<0.05),and the combined effect of prednisolone was significantly lower than that of other steroids(P<0.05).The combined effect of lipopolysaccharide 100 μg/kg×twice was significantly lower than 10 μg/kg×twice and 50 μg/kg×twice(P<0.05).The rate of empty bone lacunae in the model induced by steroid combined with serum was related to serum dose and steroid type(P<0.05),among which the combined effect amount of dexamethasone sodium phosphate was significantly higher than other steroid types(P<0.05),and the combined effect amount of dexamethasone was significantly lower than other steroid types(P<0.05);the combined effect amount of serum"10 mL/kg+6 mL/kg"combined dose was lower than other serum doses(P<0.05). CONCLUSION:(1)With the rate of empty bone lacunae and the ratio of trabecular bone area as the judgment standard for the successful establishment of the model,the three modeling methods can successfully construct the rabbit steroid-induced osteonecrosis of femoral head model,of which the steroid combined with lipopolysaccharide method is the best.(2)New Zealand white rabbits and dexamethasone are recommended when selecting the steroid-alone method.Methylprednisolone sodium succinate and low-dose lipopolysaccharide are recommended when selecting the steroid combined with lipopolysaccharide method.Dexamethasone sodium phosphate is recommended when selecting the steroid combined with serum modeling method.

Effect of ginseng polysaccharide on the expression of prostaglandin E2/6-keto-prostaglandin 1alpha in traumatic osteoarthritis model rats / 中国组织工程研究

BACKGROUND:Ginseng extracts have been found to significantly improve osteoarthritis,but the therapeutic effects of ginseng polysaccharide extracts on osteoarthritis have not been reported. OBJECTIVE:To investigate the effect of ginseng polysaccharide on the expression of prostaglandin E2/6-keto-prostaglandin F1α in traumatic osteoarthritis model rats. METHODS:Sixty male Sprague-Dawley rats were selected and randomly divided into healthy group,model group,ginseng polysaccharide low-dose group,ginseng polysaccharide medium-dose group,ginseng polysaccharide high-dose group and dexamethasone group.Except for 10 rats in the healthy group,the other rats were taken to establish traumatic osteoarthritis models.The healthy group and model group were given 0.2 mL of normal saline intraperitoneally.The low-,medium-,and high-dose groups were intraperitoneally injected with 0.1,0.25,0.5 μg/mL ginseng polysaccharide,respectively.In the dexamethasone group,0.2mg/kg dexamethasone(0.2 mL)was injected intraperitoneally.Injections were given once every 3 days,for 4 consecutive weeks.Serum prostaglandin E2 and 6-keto-prostaglandin F1α levels were detected by ELISA.The bone and joint function of rats were assessed by the Mankin's score.Hematoxylin-eosin staining was used to observe the pathologic morphology of the knee joints of rats.Western blot and PCR were used to detect the protein and mRNA expression of tumor necrosis factor α and interleukin-1β,interleukin-10 in articular cartilage tissue,respectively. RESULTS AND CONCLUSION:Compared with the model group,serum prostaglandin E2 levels were decreased in the medium-dose group and dexamethasone group,while serum 6-keto-prostaglandin F1α levels were increased(P<0.05).Compared with the medium-dose group and dexamethasone group,the above-mentioned indicators were significantly improved in the high-dose group,and there was no significant difference between the medium-dose group and dexamethasone group(P>0.05).Compared with the model group,the Mankin's score was reduced in the medium-dose group and dexamethasone group(P<0.05),but there was no significant difference between the medium-dose group and dexamethasone group(P>0.05).Compared with the medium-dose group and dexamethasone group,the Mankin's score was significantly reduced in the high-dose group(P<0.05).The cartilage tissue layer of rats in the model and low-dose groups was significantly thinned,the cracks and chondrocytes deep into the bone layer were largely lost,the tide line was seriously broken and blurred,the collagen fibers in the synovial layer were increased and thickened,and a large number of chondrocytes were destroyed and arranged irregularly.These pathological changes were improved in the medium-dose group and dexamethasone group compared with the model group as well as improved in the high-dose group compared with the medium-dose group.Compared with the model group,the expression of tumor necrosis factor-α and interleukin-1β was reduced,while the expression of interleukin-10 was increased in the medium-dose group and dexamethasone group(P<0.05).These indicators in the joint were significantly improved in the high-dose group compared with the medium-dose group and dexamethasone group(P<0.05),but there was no significant difference between the medium-dose group and dexamethasone group(P>0.05).To conclude,ginseng polysaccharide can improve the inflammatory level and pathological morphology of traumatic osteoarthritis rats and reduce the Mankin's score.Its mechanism may be related to the regulation of prostaglandin E2/6-keto-prostaglandin F1α levels.