Chemopreventive agents on a mouse skin carcinogenesis model: an integrative review / Agentes quimiopreventivos em um modelo de carcinogênese de pele em camundongos: uma revisão integrativa / Agentes quimiopreventivos en un modelo de carcinogénesis cutánea en ratón: una revisión integradora

Squamous cell carcinoma (SCC) is a non-melanoma skin cancer, with chronic sun exposure as the main risk factor. Excisional surgery is the most indicated treatment; however, patients can suffer functional, aesthetic, and psychological damage depending on the lesion site. Topical administration of 7,12-dimethylbenz[a]anthracene (DMBA) and 12-O-Tetradecanoylphorbol-13- acetate (TPA) induce to the appearance of benign skin tumors in mice, some of which develop into SCC. This protocol has been used to analyze the effects of many chemopreventive agents that may block or inhibit the mechanisms of action of chemical carcinogenesis. We compared the effects of chemopreventive agents in an induced skin carcinogenesis animal model. In the Scopus, PubMed, and EMBASE databases, we searched for manuscripts published between June 16, 2011, and June 16, 2021. We excluded studies conducted in vitro or on transgenic mice; in addition, studies without drug dosage, route of administration, or tumor incidence were excluded. We selected 26 studies and analyzed their main characteristics and the outcomes of tumorigenesis analysis. Most chemopreventive agents have shown excellent potential to inhibit the development of skin tumors. This review also discusses the standardization of studies in animal models to ensure better responses and future randomized clinical trials for cancer treatment and prevention.

O carcinoma espinocelular cutâneo (CEC) é um câncer de pele não melanoma, com a exposição solar crônica como o principal fator de risco. A cirurgia excisional é o tratamento mais indicado; entretanto, os pacientes podem sofrer danos funcionais, estéticos e psicológicos dependendo do local da lesão. A administração tópica de 7,12-dimetilbenz[a]antraceno (DMBA) e 12-O- Tetradecanoilforbol-13-acetato (TPA) induz ao aparecimento de tumores cutâneos benignos em camundongos, alguns dos quais evoluíram para CEC. Este protocolo tem sido utilizado para analisar os efeitos de muitos agentes quimiopreventivos que podem bloquear ou inibir os mecanismos de ação da carcinogênese química. Comparamos os efeitos de agentes quimiopreventivos em um modelo animal que foi induzido à carcinogênese de pele. Nas bases de dados Scopus, PubMed e EMBASE, buscamos manuscritos publicados entre 16 de junho de 2011 e 16 de junho de 2021. Excluímos estudos realizados in vitro ou em camundongos transgênicos; além disso, estudos sem dosagem de drogas, via de administração ou incidência de tumores foram excluídos. Selecionamos 26 estudos e analisamos suas principais características e os resultados da análise da tumorigênese. A maioria dos agentes quimiopreventivos tem demonstrado excelente potencial para inibir o desenvolvimento de tumores cutâneos. Esta revisão também discute a padronização de estudos em modelos animais para garantir melhores respostas e futuros ensaios clínicos randomizados para tratamento e prevenção do câncer.

El carcinoma de células escamosas (CCE) es un cáncer de piel no melanoma, cuyo principal factor de riesgo es la exposición crónica al sol. La cirugía de escisión es el tratamiento más indicado; sin embargo, los pacientes pueden sufrir daños funcionales, estéticos y psicológicos dependiendo de la localización de la lesión. La administración tópica de 7,12-dimetilbenz[a]antraceno (DMBA) y 12-O-Tetradecanoilforbol-13-acetato (TPA) inducen a la aparición de tumores cutáneos benignos en ratones, algunos de los cuales se convierten en CCE. Este protocolo se ha utilizado para analizar los efectos de muchos agentes quimiopreventivos que pueden bloquear o inhibir los mecanismos de acción de la carcinogénesis química. Comparamos los efectos de los agentes quimiopreventivos en un modelo animal de carcinogénesis cutánea inducida. En las bases de datos Scopus, PubMed y EMBASE, se buscaron los manuscritos publicados entre el 16 de junio de 2011 y el 16 de junio de 2021. Se excluyeron los estudios realizados in vitro o en ratones transgénicos; además, se excluyeron los estudios sin dosis de fármacos, vía de administración o incidencia tumoral. Se seleccionaron 26 estudios y se analizaron sus características principales y los resultados del análisis de la tumorigénesis. La mayoría de los agentes quimiopreventivos han mostrado un excelente potencial para inhibir el desarrollo de tumores cutáneos. Esta revisión también analiza la estandarización de los estudios en modelos animales para garantizar mejores respuestas y futuros ensayos clínicos aleatorios para el tratamiento y la prevención del cáncer.

Modelo preclínico de obesidad basado en alimentos altamente procesados y de alta palatabilidad / Preclinical model of obesity based on ultra-processed and highly palatable foods

Resumen Introducción: el alarmante incremento de la obesidad en todo el mundo y en Costa Rica responde principalmente a modificaciones en la composición de la dieta habitual de las personas. La presente investigación tuvo como objetivo implementar un protocolo de alimentación formulado a partir de alimentos altamente procesados y de alta palatabilidad (APAP) consumidos por la población costarricense para, luego, evaluar sus efectos en la conducta alimentaria y en parámetros biométricos y bioquímicos. Metodología: ratas adultas macho Wistar se asignaron a dos grupos. Al primero se le administró alimento estándar para roedores (grupo DC) y al segundo, alimentos APAP (grupo APAP), durante ocho semanas. Resultados: a lo largo de ese periodo, los animales expuestos a los APAP exhibieron mayor ingesta y energía, caracterizadas por un alto consumo de grasas y uno menor de proteínas y fibra; además, mostraron un incremento significativo en los diversos parámetros de obesidad (e. g., peso corporal y ganancia de peso, índice de Lee y adiposidad central) y niveles descriptivamente superiores de glucosa y triglicéridos en sangre, pero notablemente menores de colesterol total. Conclusiones: los resultados indican que una dieta basada en los alimentos APAP más frecuentes en la población costarricense es capaz de inducir hiperfagia y obesidad. Así, este modelo constituye una herramienta prometedora para ahondar en el estudio de las factores neurobiológicos y metabólicos relacionados con la obesidad por el sobreconsumo de alimentos APAP.

Abstract Introduction. The alarming increase in obesity both worldwide and in Costa Rica is mainly due to changes in the composition of the usual diet of the population. The goal of our research was to implement a feeding protocol formulated from ultra-processed and highly palatable foods (UPHP) consumed by the Costa Rican population and to evaluate the effects of the UPHP diet on eating behavior and biometric and biochemical parameters. Methods: Adult male Wistar rats were assigned to two groups. One group was given standard rodent chow (DC group) while the other group received UPHP foods (UPHP group) for eight weeks. Results: Throughout this period, animals exposed to the UPHP diet exhibited higher food and energy intake characterized by high consumption of fat and lower consumption of protein and fiber. Animals in the UPHP group also showed a significant increase in obesity parameters (e.g., body weight and bodyweight gain, Lees index, and central adiposity). Furthermore, the UPHP group had descriptively higher levels of blood glucose and triglycerides and significantly lower levels of total cholesterol. Conclusions: Our results indicate that a feeding protocol based on the most frequent food choices of the Costa Rican population is capable of inducing hyperphagia and obesity. This model constitutes a promising tool to delve into the study of the neurobiological and metabolic factors related to obesity induced by overconsumption of UPHP foods.

Vitamin E suppresses aortic ultrastructural alterations induced by toxic doses of monosodium glutamate / La vitamina E suprime las alteraciones ultraestructurales aórticas inducidas por dosis tóxicas de glutamato monosódico

SUMMARY: An association between certain food additives and chronic diseases is reported. Current study determined whether administering toxic doses of the food additive monosodium glutamate (MSG) into rats can induce aortopathy in association with the oxidative stress and inflammatory biomarkers upregulation and whether the effects of MSG overdose can be inhibited by vitamin E. MSG at a dose of (4 mg/kg; orally) that exceeds the average human daily consumption by 1000x was administered daily for 7 days to the rats in the model group. Whereas, rats treated with vitamin E were divided into two groups and given daily doses of MSG plus 100 mg/ kg vitamin E or MSG plus 300 mg/kg vitamin E. On the eighth day, all rats were culled. Using light and electron microscopy examinations, a profound aortic injury in the model group was observed demonstrated by damaged endothelial layer, degenerated smooth muscle cells (SMC) with vacuoles and condensed nuclei, vacuolated cytoplasm, disrupted plasma membrane, interrupted internal elastic lamina, clumped chromatin, and damaged actin and myosin filaments. Vitamin E significantly protected aorta tissue and cells as well as inhibited MSG-induced tissue malondialdehyde (MDA), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). The highest used vitamin E dosage was more effective. Additionally, a significant correlation was observed between the aortic injury degree and tissue MDA, TNF-α, IL-6, and superoxide dismutase (SOD) levels (p=0.001). Vitamin E effectively protects against aortopathy induced by toxic doses of MSG in rats and inhibits oxidative stress and inflammation.

RESUMEN: Se reporta una asociación entre ciertos aditivos alimentarios y enfermedades crónicas. El objetivo de este estudio fue determinar si la administración de dosis tóxicas del aditivo alimentario glutamato monosódico (MSG) en ratas puede inducir aortopatía en asociación con el estrés oxidativo y la regulación positiva de los biomarcadores inflamatorios y si el efecto de una sobredosis de MSG se puede inhibir con vitamina E. Se administró MSG diariamente durante 7 días una dosis de (4 g/kg; por vía oral) que excede el consumo diario humano promedio, en 1000x a las ratas del grupo modelo. Mientras que las ratas tratadas con vitamina E se dividieron en dos grupos y se administraron dosis diarias de MSG más 100 mg/kg de vitamina E o MSG más 300 mg/kg de vitamina E. Todas las ratas fueron sacrificadas en el octavo día. Usando exámenes de microscopía óptica y electrónica, se observó una lesión aórtica profunda en el grupo modelo demostrada por una capa endotelial dañada, células musculares lisas degeneradas (SMC) con vacuolas y núcleos condensados, citoplasma vacuolado, membrana plasmática rota, lámina elástica interna interrumpida, cromatina agrupada y filamentos de actina y miosina dañados. La vitamina E protegió significativamente el tejido y las células de la aorta, además de inhibir el malondialdehído tisular (MDA) inducido por MSG, la interleucina-6 (IL-6) y el factor de necrosis tumoral alfa (TNF-α). La dosis más alta de vitamina E utilizada fue más efectiva. Además, se observó una correlación significativa entre el grado de lesión aórtica y los niveles tisulares de MDA, TNF-α, IL-6 y superóxido dismutasa (SOD) (p=0,001). La vitamina E efectivamente protege contra la aortopatía inducida por dosis tóxicas de MSG en ratas e inhibe el estrés oxidativo y la inflamación.

Ambystoma mexicanum, A MODEL ORGANISM IN DEVELOPMENTAL BIOLOGY AND REGENERATION: A COLOMBIAN EXPERIENCE / Ambystoma mexicanum, un organismo modelo en biología del desarrollo y regeneración: experiencia colombiana

ABSTRACT Ambystoma mexicanum is a urodele amphibian endemic to Xochimilco Lake in Mexico, it belongs to the salamander family Ambystomatidae. This species has frequently been used as model organism in developmental biology and regeneration laboratories around the world due to its broad regenerative capacities and adaptability to laboratory conditions. In this review we describe the establishment of the first colony of axolotls in Colombia to study tissue regeneration and our perspectives on the use A. mexicanum as a model organism in Colombia are discussed emphasizing its possible uses in regeneration and developmental biology.

RESUMEN Ambystoma mexicanum es un anfibio urodelo endémico del lago Xochimilco en México, perteneciente a la familia de salamandras Ambystomatidae. Esta especie se ha empleado frecuentemente como organismo modelo en laboratorios de biología del desarrollo y regeneración alrededor del mundo, dadas sus amplias capacidades regenerativas y adaptabilidad en condiciones de laboratorio. En esta revisión, se describe el establecimiento de la primera colonia de ajolotes en Colombia, para adelantar estudios de regeneración de tejidos, y se discuten las perspectivas de A. mexicanum como organismo modelo en el país, enfatizando sus posibles usos en regeneración y biología del desarrollo.

Healing of Wounds Treated with Chitosan Hydrogels with Extracts from Aloe vera and Calendula officinalis / Cicatrización de Heridas Tratadas con Hidrogeles de Quitosano con Extractos de Aloe vera y Calendula officinalis

ABSTRACT This project's purpose was to evaluate the healing effects of chitosan (CS) hydrogels loaded with extracts from Aloe vera (CS+AV) and Calendula officinalis (CS+CO) on wounds of diabetic and non-diabetic Wistar rats. A total of 24 rats were used; animals were randomly divided into three diabetic and three non-diabetic groups (one control and two treated groups) and monitored for 13 days. A biopsy on the wound site was recovered to assess the collagen and n-acetyl glucosamine content. The wound area ratio was reduced since day 1 on both non-diabetic treated groups. A similar effect was observed on the diabetic group treated with CS+AV, while the diabetic group treated with CS+CO showed a reduction in wound area compared to the diabetic control until day 11 after being wounded. Collagen and n-acetyl glucosamine content were higher in every treated group. Further studies are needed to clarify the underlying mechanisms through which they promote wound healing. These results suggest that the hydrogels prepared are potential material to be used as wound dressings.

RESUMEN El propósito de este proyecto fue evaluar los efectos curativos de los hidrogeles de quitosano con extractos de Aloe vera (CS + AV) y Calendula officinalis (CS + CO) en heridas en ratas Wistar diabéticas y no diabéticas. Se utilizaron un total de 24 ratas; los animales fueron divididos aleatoriamente en tres grupos diabéticos y tres no diabéticos (un grupo control y dos tratados) y se monitorearon durante 13 días. Se recuperó una biopsia del sitio de la herida para evaluar el contenido de colágeno y n-acetilglucosamina. El área de la herida se redujo desde el día 1 en ambos grupos no diabéticos tratados. Se observó un efecto similar en el grupo diabético tratado con CS + AV, mientras que el grupo diabético tratado con CS + CO mostró una reducción del área de la herida en comparación al control diabético hasta el día 11 después de la creación de la herida. El contenido de colágeno y n-acetilglucosamina fue mayor en todos los grupos tratados. Se necesitan más estudios para aclarar los mecanismos subyacentes a través de los cuales estos tratamientos promueven la cicatrización de heridas. Estos resultados sugieren que los hidrogeles preparados son materiales con potencial para usarse como apósitos para heridas.

Hematological Changes after Caprine Demineralized Bone Matrix Implantation in Ulnar Critical Defect of Rabbit Model / Alterações hematológicas após implante de matriz óssea desmineralizada caprina em modelo de defeito ulnar crítico em coelhos

Abstract Objective Several animal models have been used in fracture healing and bone graft studies, but hematological responses are seldom reported. Therefore, the present study reported the hematological changes observed in rabbits that underwent xenografting of caprine demineralized bone matrix (CDBM). Method Twenty-four (24) male rabbits (2.5 0.5kg) were acquired for the purpose of this study and were randomly assigned to three groups: autologous bone graft (ABG), unfilled (NC), and caprine demineralized bone matrix (CDBM). Blood samples were collected through cardiac puncture under xylazine-ketamine anesthesia on day 0 (baseline), and on days 28 and 56 postsurgery and were analyzed manually within 2hours of collection. Statistical analysis was performed using a two-way analysis of variance (ANOVA) with repeated measures, and a p-value< 0.05 was considered significant. Result There was an overall significant difference in the values of total white blood cell count (p» 0.0043), neutrophil count (p< 0.0001), monocyte count (p» 0.0184), red blood cell count (p» 0.003), hemoglobin concentration (p< 0.0001) and packed cell volume (p< 0.0001) across the days and the treatment groups. There was, however, no overall significant difference in lymphocyte count (p» 0.4923), basophil count (p» 0.4183), and eosinophil count (0.4806) within days. Conclusion Response to CDBM grafting in rabbits could, therefore, be said to be characterized by marked leukocytosis with neutrophilia, lymphocytosis, and monocytosis by day 28 of postgrafting. This could form the basis with which hematology can be used to monitor body response of bone graft animal models.

Resumo Objetivo Diversos modelos animais têm sido usados em estudos sobre enxertos ósseos e o tratamento de fraturas, mas as respostas hematológicas são raramente relatadas. Este estudo descreveu as alterações hematológicas observadas em coelhos submetidos a xenoenxertos de matriz óssea desmineralizada caprina (MODC). Métodos Vinte e quatro (24) coelhos machos (2,5 0,5 kg) foram adquiridos para este estudo e divididos aleatoriamente em três grupos: enxerto ósseo autólogo (EOA); controle negativo sem preenchimento (SP) e matriz óssea desmineralizada caprina (MODC). Amostras de sangue foram coletadas por punção cardíaca sob anestesia com xilazina-quetamina no dia 0 (para estabelecimento dos valores basais) e aos dias 28 e 56 após a cirurgia; essas amostras foram submetidas à análise manual em até 2 horas após a coleta. A análise estatística foi composta por análise de variância (ANOVA) de dois fatores com medidas repetidas, e o valor de p< 0,05 foi considerado significativo. Resultados Houve uma diferença geral significativa nos números de leucócitos totais (p» 0,0043), neutrófilos (p< 0,0001), monócitos (p» 0,0184) e hemácias (p» 0,003), na concentração de hemoglobina (p< 0,0001) e no hematócrito (p< 0,0001) ao longo dos dias e entre os grupos de tratamento. No entanto, não houve diferença global significativa no número de linfócitos (p» 0,4923), basófilos (p» 0,4183) e eosinófilos (p» 0,4806) entre os dias. Conclusão A resposta ao enxerto de MODC em coelhos é, portanto, caracterizada por leucocitose intensa com neutrofilia, linfocitose e monocitose no 28° dia após o procedimento. Esses dados podem basear a utilização da hematologia no monitoramento da resposta corporal em modelos animais de enxerto ósseo.

Effects of laser photobiomodulation on tgf-ß and vegf expression in burn wound: systematic review and meta-analysis in the animal model / Influencia de la fotobiomodulación por láser en la expresión de tgf-ß y vegf en heridas por quemadura: revisión sistemática y meta-análisis en modelo animal

SUMMARY: Laser photobiomodulation (laser PBM) is known to be able to accelerate burn wound healing in the animal model; however little evidence exists on the action of laser PBM on the expression of important proteins in wound healing in the animal model, such as VEGF and TGF-ß1. The aim of this study was to carry out a systematic review in order to analyse the effect of laser PBM on VEGF and TGF-ß expression during burn wound repair in the animal model. A systematic review was carried out of the EMBASE, PubMed/ MEDLINE and LILACS databases. The studies included were preclinical studies that analysed the action of laser PBM on the expression of VEGF and TGF-ß (1, 2, 3) during burn wound repair in the animal model. The SYRCLE risk of bias tool was used. Random effect models were used to estimate the combined effect. Increased VEGF expression was observed with the use of laser PBM at 4.93 J/cm2 per point in the first two weeks after induction of the burn wound, with greater size of effect in the second week (SDM = 5.72; 95% CI: 3.14 to 8.31, I2 = 0 %; very low certainty of evidence). We also observed that the effect of laser PBM on TGF-ß1 expression was greater than in the control in the first week (SDM = -0.45; 95% CI: -1.91 to 1.02, I2 = 51 %; very low certainty of evidence), but diminished in the third week after induction of the lesion (SDM = -2.50; 95% CI: 3.98 to -1.01, I2 = 0 %; very low certainty of evidence). Laser PBM has an effect on TGF-ß1 and VEGF expression, promoting burn wound repair in the animal model.

RESUMEN: Es sabido que la fotobiomodulación por láser (FBM láser) puede acelerar el proceso de curación de heridas por quemadura en modelo animal, sin embargo aún se carece de mayor evidencia sobre la acción de la FBM láser en la expresión de proteínas importantes en el proceso de curación de heridas en modelo animal, como VEGF y TGF-ß1. Así, el objetivo de este estudio fue realizar una revisión sistemática a fin de analizar el efecto de la FBM láser sobre la expresión de VEGF, TGF-ß durante el proceso de reparación de heridas por quemadura en modelo animal. Se realizó una búsqueda sistemática en las bases de datos EMBASE, PubMed/MEDLINE y LILACS. Se incluyeron estudios preclínicos que analizaron la acción de la FBM láser en la expresión de VEGF, TGF-ß (1, 2, 3) durante el proceso de reparación de heridas por quemadura en modelo animal. Se utilizó la herramienta de riesgo de sesgo SYRCLE. Se utilizaron modelos de efectos aleatorios para estimar el efecto combinado. Observamos aumento de la expresión de VEGF con el uso de FBM láser 4.93 J/cm2 por punto, en las dos primeras semanas tras inducción de la herida por quemadura, con mayor tamaño de efecto en la segunda semana (SDM = 5,72; IC del 95%: 3,14 a 8,31, I2 = 0 %; certeza de la evidencia muy baja). También se observó el efecto de la FBM láser en la expresión del TGF- ß1 que fue mayor que el control en la primera semana (SDM = - 0,45; IC del 95%: -1,91 a 1,02, I2 = 51 %; certeza de la evidencia muy baja), disminuyendo en la tercera semana tras inducción de la lesión (SDM = -2,50; IC del 95%: -3,98 a -1,01; I2 = 0 %; certeza de la evidencia baja). La TFB por láser ejerce influencia en la expresión de TGF-ß1 y VEGF favoreciendo el proceso de reparación de heridas por quemadura en modelo animal.

Research progress on animal model of mixed infection of microorganisms keratitis / 国际眼科杂志(Guoji Yanke Zazhi)

@#Microbial mixed infectious keratitis is an ocular surface disease caused by corneal infection, which has an acute onset and rapid progression and can lead to blindness in severe cases. Establishing an animal model of microbial mixed infectious keratitis is conducive to exploring its pathogenesis, prevention, clinical diagnosis and treatment. This article reviews the methods of making animal models of mixed infectious keratitis with microorganisms and the diagnostic methods after successful modelling infections, aiming to provide references for the further development and research of animal models of the disease.

Comparative study on establishing an immune tolerance model for allergic conjunctivitis in mice with different methods / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To establish an immune tolerance model for allergic conjunctivitis in newborn mice with different methods and observe the impact of environmental factors on allergic conjunctivitis in early life.METHOD: A total of 50 Balb/c newborn mice were randomly divided into blank control group, ovalbumin(OVA)+subcutaneous injection group, OVA+nebulized inhalation group, OVA+gastric group, ragweed pollen(RW)+subcutaneous injection group, RW+nebulized inhalation group, RW+gastric group, house dust mite(HDM)+subcutaneous injection group, HDM+nebulized inhalation group, HDM+intragastric group(n=5 animals/group). Except for the blank control group, mice in each group were individually exposed to the corresponding antigens to induce immune tolerance early in life and stimulated with the corresponding antigens in adulthood. The ocular surface was visualized by anterior segment photography. The relative expression level of conjunctival RANTES and IL-17 mRNA was measured by RT-qPCR and serum IL-17 concentration was measured by ELISA.RESULTS: Compared with the blank control group, the relative expression level of conjunctiva IL-17 mRNA in RW+gastric group was the highest, and it was the lowest in RW+subcutaneous group(all P&#x003C;0.05). The relative expression level of conjunctiva RANTES mRNA was the highest in RW+gastric group(P&#x003C;0.001). Compared with the blank control group, the serum concentration of IL-17 was increased in all treatment groups except OVA+nebulizer group and RW+subcutaneous group(P&#x003C;0.05).CONCLUSION: The immune tolerance of allergic conjunctivitis induced by subcutaneous injection of antigen was the most suitable method in the early life of mice.

Establishment of an animal model of Sparganum mansoni infection and study on therapeutic methods II Establishment of a mouse model of sparganosis mansoni via oral administration of procercoids / 中国血吸虫病防治杂志

Objective To establish an animal model of sparganosis mansoni through oral administration of Cyclops infected with procercoids. Methods Domestic cats were infected with Sparganum mansoni under laboratory conditions, and fresh cat stool samples were collected, washed in dechlorinated water, and filtered. Spirometra mansoni eggs were collected and prepared into suspensions. Twenty C57BL/6j mice were randomly divided into the experimental group (n = 15) and the control group (n = 5). Wild Cyclops were infected with Spirometra mansoni coracidia to allow 3 to 5 procercoids in each Cyclop. Then, each mouse in the experimental group was given 15 Cyclops infected with procercoids by gavage, while mice in the control group were orally administered with the same volume of dechlorinated water. All mice were sacrificed after 5 months, and dissected, and suspicious Sparganum mansoni worms were collected. The serum specific IgG antibody against Sparganum mansoni was measured in mice using enzyme-linked immunosorbent assay (ELISA). Genomic DNA was isolated from suspicious Sparganum mansoni worms, and the specific Sparganum mansoni cytochrome oxidase I (COI) gene was amplified using PCR assay. Results Among the 15 mice in the experimental group, six were positive for the serum specific IgG antibody against Sparganum mansoni, and milky white worms were found and collected from the subcutaneous regions of 4 out of 6 mice. Only one worm was detected in each mouse, and the worm morphology was similar to Sparganum mansoni. Capillary electrophoresis of the PCR amplification products of COI gene presented a specific band with 151 bp in size, and sequencing analysis revealed 100% homology with Sparganum mansoni. Conclusions A mouse model of sparganosis mansoni is successfully created through oral administration of Cyclops infected with Spirometra mansoni procercoids.

Establishment and Evaluation of Animal Model of Oligoasthenospermia / 中国实验方剂学杂志

Oligozoospermia and asthenospermia are common causes of clinical male infertility. The number of patients increases year by year and shows a younger trend. Its etiology is complex， the mechanism and unclear pathogenesis and rely on empirical therapy. Therefore， it is necessary for exploring the pathogenesis and developing corresponding drugs to establish reasonable animal models. By comparing different animal model making methods， this paper provides ideas for constructing a more standardized animal model of oligoasthenospermia. At the moment， a lot of molding methods for oligoasthenospermia are available. Combined with the animal experimental articles of oligoasthenospermia in recent years， this study described the modeling with adenine， ornidazole， tripterygium glycoside， hydrocortisone， cyclophosphamide， busulfan， paclitaxel， heat stress， ionizing radiation， high-fat diet， and gene knockout， respectively， and compared the modeling methods in terms of the time， indexes， animal line， and model evaluation. Thereby， the advantages and disadvantages of different models of oligoasthenospermia were summarized， and finds that the existing animal models of oligoasthenospermia still have many shortcomings that need to be further improved. The selection， standardization and innovation of animal models need to be solved urgently， and the coincidence between animal models and clinical patients' traditional Chinese medicine syndromes is not coincident. In view of the existing problems， we should further explore how to build a modeling method in line with clinical characteristics and syndrome types， select the compound model method of integrated traditional Chinese and Western medicine， copy the model closer to the law of disease development and in line with traditional Chinese medicine syndrome， and provide animal experimental support for exploring the mechanism of disease， developing characteristic drugs and guiding clinical medication.

Establishment of A Mouse Model of Obesity and Depression Comorbidity and Its Application in Pharmacological Study of Tripterine / 中国实验方剂学杂志

ObjectiveTo establish a neuroinflammation-based obesity and depression comorbidity （COM） model in mice and explore the pharmacodynamics and preliminary pharmacological mechanism of tripterine on COM mice. MethodC57BL/6J mice were randomly divided into a normal group （Chow）， a diet-induced obesity group （DIO）， and a COM group. The mice in the COM group were fed on a high-fat diet and chronically stressed with moist litter for 12 weeks to establish the COM model. C57BL/6J mice were randomly divided into a Chow group， a COM group， and a tumor necrosis factor-α（TNF-α） knock-down group. In the TNF-α knock-down group， TNF-α shRNA adeno-associated virus was injected into the amygdala through brain stereotaxis， and the expression of TNF-α in the amygdala was down-regulated. C57BL/6J mice were randomly divided into a Chow group， a DIO group， a DIO + low-dose tripterine group （0.5 mg·kg-1）， a DIO + high-dose tripterine group （1.0 mg·kg-1）， a COM group， a COM + low-dose tripterine group （0.5 mg·kg-1）， and a COM + high-dose tripterine group （1.0 mg·kg-1）. The body weight， food intake， glucose tolerance， white/brown fat ratio， serum total cholesterol （TC）， triglyceride （TG）， and high-/low-density lipoprotein cholesterol （HDL-C and LDL-C） content were recorded， and obesity of mice in each group was evaluated. Forced swimming test （FST）， tail suspension test （TST）， and open field test were used to evaluate the degree of depression of mice in each group. Immunofluorescence staining was used to detect the protein expression levels of neuropeptide Y， tryptophan hydroxylase 2 （TPH2）， and brain-derived neurotrophic factor （BDNF） in various brain nuclei of mice. Correlation analysis was used to detect the correlation of obesity and depression indexes. ResultThe comparison of the Chow group and the DIO group indicated that COM mice showed obesity and depression. To be specific， obesity was manifested as increased body weight and food intake （P<0.05， P<0.01）， as well as increased NPY expression in the central amygdala， and depression was manifested as prolonged immobility time in FST and TST （P<0.01）， and reduced TPH2-positive 5-hydroxytryptamine neurons in the dorsal raphe nucleus （DRN） and basolateral nucleus of the amygdala （BLA）. The down-regulation of TNF-α protein in BLA of COM mice shortened the immobility time in FST and TST （P<0.05， P<0.01）， increased TPH2/BDNF-positive neurons in BLA， and showed no significant changes in obesity. In DIO mice， the administration of 0.5 mg·kg-1 tripterine for 9 days significantly decreased the 60 min blood glucose in glucose tolerance （P<0.01） and food intake （P<0.05）. In COM mice， 1.0 mg·kg-1 tripterine was administered for 14 days to significantly decrease 30 min blood glucose in glucose tolerance （P<0.01）， and food intake （P<0.05）， and immobility time in TST （P<0.01）， increase TPH2-BDNF double-labeled cells in BLA and DRN， and reduce the area of TMEM119-stained cells. ConclusionThe model of obesity and depression comorbidity can be properly induced in mice under the condition of dual stress of energy environment. Tripterine can effectively interfere with obesity-depression comorbidity， and its mechanism may be related to the inhibition of central nervous system inflammation.

Animal Model of Cold Congeal and Blood Stasis Syndrome： A Review / 中国实验方剂学杂志

The cold congeal and blood stasis syndrome is a common clinical traditional Chinese medicine（TCM） syndrome. The animal model of cold ongeal and blood stasis syndrome is the basis for exploring the essence of TCM cold congeal and blood stasis syndrome，and the premise of follow-up TCM clinical research.This paper summarized the preparation method， theoretical support，and evaluation method of animal models of cold congeal and blood stasis syndrome in recent years and analysed the strengthens and weaknesses of different models. At present，the common animal models of cold congeal and blood stasis syndrome mainly include etiological model，etiological and pathological composite model and disease-syndrome combination model. The etiological model was mainly prepared by cold exposure，which could be divided into whole-body freezing， ice bath and local frostbite. The etiological and pathological composite model was mainly prepared by cold stimulation combined with epinephrine injection. The common disease-syndrome combination models included the coronary heart disease model of cold congeal and blood stasis syndrome，primary dysmenorrhea model of cold congeal and blood stasis syndrome，endometriosis model of cold congeal and blood stasis syndrome， and arteriosclerosis obliterans model of cold congeal and blood stasis syndrome. The three models have both advantages and disadvantages. Specifically， the disease-syndrome combination model had the highest consistency with clinical practice and was more reliable and practical. However， the disease types of this model were specific，and the combination method of disease and syndrome was controversial. The evaluation indicators of the animal models of cold congeal and blood stasis syndrome focused on the characterization of the syndrome and the physico-chemical indicators related to blood flow，such as blood rheology，coagulation function and microcirculation. In addition， some scholars explored the evaluation indicators from the aspects of vasomotor function，endocrine and energy metabolism. The objectivity and specificity of the current model evaluation methods needed to be further improved. The research of animal model of cold congeal and blood stasis syndrome should be based on clinical practice and oriented by clinical demand. Only by establishing animal models that are highly consistent with the characteristics of clinical disease and syndrome can we better reveal the essence of cold congeal and blood stasis syndrome and promote the modernization of TCM.

Establishment and Evaluation of Ulcerative Colitis Animal Model in Combination of Disease and Syndrome / 中国实验方剂学杂志

Ulcerative colitis （UC） is one of the main manifestations of inflammatory bowel disease. Because of its lingering and refractory nature， it has become a major public health challenge worldwide. In the treatment of UC， traditional Chinese medicine （TCM） effectively relieves clinical syndromes， shortens the treatment period， reduces the frequency of recurrence， improves the quality of life， and reduces the occurrence of complications. To study the specific mechanism of TCM in the treatment of UC and screen out suitable drugs under the guidance of syndrome differentiation， the suitable UC animal model in the combination of disease and syndrome is used as an important method. This paper summarized and compared the UC animal model in the combination of disease and syndrome from five aspects， including selection of model animal species， sexual selection， preparation methods of UC animal model in the combination of disease and syndrome， indicators of model evaluation， and the main mechanism of TCM intervention in UC animal model in the combination of disease and syndrome. This paper aimed to provide references for the establishment of the optimal UC animal model in the combination of disease and syndrome. Research shows that UC syndrome mainly studied at present includes damp-heat syndrome， spleen deficiency syndrome， spleen-kidney Yang deficiency syndrome， spleen deficiency and dampness accumulation syndrome， liver depression and spleen deficiency syndrome， and cold-heat mixed syndrome.In the modeling method， the etiology simulation method is mainly used to first copy the syndrome type before the chemical agents or immune preparations were used to induce the disease model，and rats were often selected as the research objects，and the replication cycle was 7 to 28 days.The selected chemical reagents were mainly 5% dextran sulfate sodium（DSS） free drink, 2,4,6-trinitrobenzenesulfonic acid（TNBS） 100 mg·kg-1 and 50% ethanol 0.25 mL mixed reagent enema.This model replication method can take into account both UC pathogenesis characteristics of pathology of western medicine and TCM， syndrome type of traditional Chinese medicine and western medicine for interpretation pathological changes and TCM treatment of UC associated mechanism is of great significance， and help to help toestablish the optimal condition in combination with UC animal models for reference， for further research on prevention and treatment of UC specific mechanism of action of TCM model basis.

Animal Model Analysis of Dental Arthritis Based on Clinical Syndrome Characteristics of Traditional Chinese Medicine and Western Medicine / 中国实验方剂学杂志

In recent years， as people's diets have changed and diversified， the incidence of dental arthritis has increased year by year， seriously affecting the quality of life of patients. Therefore， the prevention and treatment of dental arthritis should be emphasized. To further study the pathogenesis of dental arthritis and the development and screening of therapeutic drugs， this paper summarized and analyzed the modeling methods， mechanisms， as well as the advantages and disadvantages of the existing animal models of dental arthritis. The clinical diagnostic criteria of traditional Chinese medicine （TCM） and western medicine was established， and the compatibility of TCM and western medicine anastomosis in animal models was evaluated. The results showed that the gel perfusion model had a good match between TCM and western medicine， with simple operation and short cycle. By combining the pathogenic factors of TCM and western medicine， the models of kidney deficiency and stomach heat with kidney deficiency in TCM were obtained， which fully reflected the clinical syndrome characteristics of TCM and western medicine， thus simulating the pathogenesis of human natural dental arthritis. Besides， ligation line model， as the most commonly-used animal model of dental arthritis with a good match to western medicine， was mature and highly repeatable， and had a high success rate. Ligation line model was widely used in various periodontal disease studies， but it did not involve the pathogenic factors of TCM. The current animal model of dental arthritis is given priority to western medicine disease model， and the combination of disease and model is rare， which cannot meet the requirements of the syndrome characteristics of TCM. Only an animal model of dental arthritis with TCM syndrome that conforms to the clinical syndrome characteristics effectively assists to study the nature of TCM syndrome and develop innovative Chinese medicine. Therefore， the establishment of an accurate and standardized animal model of dental arthritis combined with TCM and western medicine is still the focus of future study on the pathogenesis of dental arthritis. This study is intended to provide a certain basis for the discovery， screening， and evaluation of medicines for the treatment of dental arthritis.

Mechanism of Chaihu Qinggantang in Intervening NLRP3/IL-1β Pathway to Treat Granulomatous Lobular Mastitis in Rat Model / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Chaihu Qinggantang （CHQGT） in the treatment of granulomatous lobular mastitis （GLM） in the rat model. MethodSixty female rats were divided into a normal group， a model group， a prednisolone group （0.001 8 g·kg-1）， and three CHQGT low-dose， medium-dose， and high-dose groups （4.5， 8.9， 17.8 g·kg-1）. The tissue homogenates mixed with GLM lesion tissue and Fritner's reagent were used for modeling. After modeling， the treatment groups were given corresponding treatment factors， and the normal group and the model group were given the equal volume of normal saline. The changes in mammary gland of rats were observed after 14 d. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in breast samples. The mRNA expressions of NOD-like receptor thermal protein domain associated protein 3 （NLRP3） inflammasome， Caspase-1， and interleukin-1β （IL-1β） were detected by real-time quantitative fluorescence polymerase chain reaction （Real-time PCR）. The protein expressions of NLRP3， Caspase-1， IL-1β， and IL-18 were detected by Western bolt. ResultAs compared with the normal group， the breasts of rats in the model group were obviously swelling， and mammary gland inflammation index was significantly increased （P<0.01）. Pathological changes included the formation of granuloma centered on the lobule of mammary gland with a large number of inflammatory cells such as lymphocytes and plasma cells. The mRNA expressions of NLRP3， Caspase-1， and IL-1β， and the protein expressions of NLRP3， Caspase-1， IL-1β， and IL18 in the model group were significantly increased （P<0.01）. Compared with the model group， the treatment groups improved breast swelling， and the CHQGT medium and high-dose groups and the prednisolone group reduced inflammation index to some extent after treatment （P<0.05， P<0.01）. The inflammation degree of mammary gland was significantly improved， and inflammatory cells such as macrophages， lymphocytes， and plasma cells were reduced to varying degrees in pathological aspects. The mRNA expressions of NLRP3， Caspase-1， and IL-1β， and the protein expressions of NLRP3， Caspase-1， IL-1β， and IL-18 in the CHQGT high-dose group and the prednisolone group were significantly down-regulated （P<0.05， P<0.01）. ConclusionCHQGT inhibits inflammation and treats GLM in rats. The mechanism is possibly related to the inhibition of NLRP3/IL-1β signaling pathway， which provides a new target for the prevention and treatment of GLM by Qingxiao method.

Animal Model of Esophageal Cancer Based on Its Clinical Symptoms of Traditional Chinese and Western Medicine / 中国实验方剂学杂志

ObjectiveEsophageal cancer is a common malignant tumor of the upper gastrointestinal tract， and has high incidence and mortality in China. Its incidence is increasing year by year， and survival rate is low， thus seriously threatening human life and health. To further explore the pathogenesis of esophageal cancer and its systematic and efficient diagnosis and treatment methods， the animal models of esophageal cancer was evaluated according to the animal model evaluation method previously established by our team based on the characteristics of its clinical symptoms of traditional Chinese and western medicine， and suggestions for model improvement were proposed. MethodThe existing animal models of esophageal cancer were summarized through China National Knowledge Infrastructure （CNKI） and Wanfang Data. The relevant indexes of the models were assigned， and their coincidence with the clinical diagnostic guidelines of traditional Chinese and western medicine for esophageal cancer was evaluated. ResultExcept the spontaneous animal model of esophageal cancer with high clinical coincidence adopted in few studies， the animal model induced by methylbenzylnitrosamine was in good agreement with the clinical data， which simulated the etiology and pathogenesis of esophageal cancer to a certain extent. The model partially reflected some indicators of clinical diagnosis in western medicine， and also indicated the body weight loss， purple nail and increased number of drinking in traditional Chinese medicine （TCM）. However， there was still a lack of differentiation of TCM syndromes. ConclusionOn the basis of the original model， the animal model induced by methylbenzylnitrosamine and the mouse model of xenogeneic gastric wall transplantation of human esophageal cancer cells were applied， which were intervened with the factors of phlegm and qi mutual obstruction syndrome， blood stasis and phlegm stagnation syndrome， Yin deficiency and internal heat syndrome and Qi deficiency and yang weakness syndrome， and were distinguished to reflect the performance of TCM syndrome. The animal model of esophageal cancer combined with TCM syndrome was thus obtained， which embodied the clinical symptoms of esophageal cancer in TCM， and the characteristics of the animal model combined with TCM syndrome， and simulated the clinical practice of traditional Chinese and western medicine， providing an accurate pathological model carrier for basic research.

Animal Model Analysis of Multiple Sclerosis Based on Clinical Characteristics of Traditional Chinese and Western Medicine / 中国实验方剂学杂志

Based on the clinical characteristics of multiple sclerosis （MS） in traditional Chinese medicine （TCM） and western medicine and literature analysis， this paper aims to formulate the diagnostic criteria of TCM and western medicine for MS. Moreover， the modeling methods of experimental autoimmune encephalomyelitis （EAE）， animals for the modeling， and characteristics of the models were analyzed and summarized， and the consistency between the EAE models and the diagnostic criteria of TCM and western medicine was evaluated. The results showed that animal models had low consistency with the clinical characteristics in TCM （highest consistency 68%） and western medicine （highest consistency 60%）. Pathological models account for the majority of animal models for MS research， but there is a lack of intuitive performance indicators. Thus， it is difficult to comprehensively evaluate the models. The mental state， limb numbness， lack of strength， loss of muscle tone， tremor， and balance disorders of the mice are among the diagnostic criteria in western medicine. In TCM diagnostic criteria， the major symptoms which are reflected in animal behavior， such as physical fatigue， lack of strength， mental fatigue， distinclination to talk， and weak heavy numb limbs， are consistent with the western diagnostic criteria. The minor symptoms， including mental decline， bitter taste in mouth， frequent and urgent urination， fecal incontinence， and aggravated fever， are not well reflected in the models. According to TCM， MS is caused by deficiency of kidney essence and external contraction of pathogen， but no index is available for evaluating the external contraction of pathogen in existing animal models. The key to experimental research on MS is to establish an appropriate animal model based on the clinical pathogenesis and characteristics. However， there is a lack of MS animal model with TCM characteristics for syndrome classification. Therefore， renewed efforts should be made to prepare animal models with both TCM and western medicine characteristics that can be used in both basic experiments and clinical research.

Animal Models of Gastric Ulcer Based on Characteristics of Clinical Symptoms in Traditional Chinese and Western Medicine：A Review / 中国实验方剂学杂志

In recent years， with the changes of people's life rhythm and living environment， the incidence of gastric ulcer has shown an increasing trend year by year， and the affected population has become younger and younger. In order to further explore the pathogenesis of gastric ulcer and its diagnosis and treatment methods， a variety of animal models of gastric ulcer have been established clinically， such as stress type， chemical factor type， pyloric ligation type， helicobacter infection type and disease-syndrome combination type. The authors intend to summarize the modeling methods and advantages and disadvantages of existing models on the basis of reviewing the etiology， pathogenesis and diagnostic criteria of gastric ulcers. It was found that the non-injurious stress method （restraint stress， restraint immersion stress and restraint freezing stress， etc.）+traditional Chinese medicine （TCM） syndrome modeling， acetic acid gavage method+TCM syndrome modeling were ideal choices for replicating animal models of acute and chronic gastric ulcer. At the same time， the analysis of the coincidence degree between each gastric ulcer model and the clinical disease characteristics of Chinese and western medicine showed that the coincidence degree of western medicine diagnostic criteria was higher than that of TCM diagnostic criteria. The successful judgment of the model was also based on western medicine diagnosis. In short， the model is insufficient in depth and breadth. It only detects a few core indicators and main indicators， ignoring the impact of secondary indicators on the diagnosis of the disease. There is also a big gap between the disease-syndrome combination model and the TCM clinical syndromes of this disease. Therefore， the depth and width of the model evaluation criteria should be strengthened， and the evaluation system of the disease-symptom combination model should be improved， in order to provide a more accurate reference for the replication of gastric ulcer models， and to replicate animal models of gastric ulcer with high coincidence degree of Chinese and western medicine for research purposes.

Research Progress of Digestive System Disease and Syndrome Combined Animal Model and Traditional Chinese Medicine Target Intervention： A Review / 中国实验方剂学杂志

Reflux esophagitis （liver-stomach disharmony， Spittoon-Qi interties， Qi and blood stasis syndrome， turbid poison intrinsic） and nonalcoholic fatty liver disease （liver depression， spleen deficiency， phlegm and blood stasis syndrome， hot and humid embodiment， phlegmy wet resistance） and functional dyspepsia （liver depression syndrome， liver stomach with spleen deficient， spleen deficiency cold syndrome， in a word， fever） is a common disease and frequently encountered disease of digestive system. The course of disease is prolonged and the prevalence is high. The successful establishment of animal model combining disease and syndrome is the premise of exploring the mechanism of traditional Chinese medicine（TCM） effect and the foundation of the development of new preparations. At the same time， mastering the complex relationship network among disease， syndrome and prescription is the prerequisite of effective treatment. When the same syndrome occurs between different diseases， the concept of "treating different diseases with the same treatment" in TCM suggests that methods can be cross-referenced for the shortage of some syndrome models. TCM intervention of digestive diseases has the characteristics of multi-path， multi-target， multi-dimension and multi-level. Therefore， this article through the literature review， summarizes the reflux esophagitis， nonalcoholic fatty liver disease and functional dyspepsia is a common disease such as the combined operation method of the model and the intervention mechanism of TCM， so as to diseases of the digestive system of different syndrome types provides the theory basis for the objective of research， and the basic research of TCM prescription and achievements provide methodological guidance.