Dietary habits of lesser bandicoot rat (Bandicota Bengalensis) in an agro-ecosystem, Pothwar Plateau, Pakistan

Abstract Dietary habits of bandicoot rats (bandicota bengalensis) were investigated in the agricultural crops of the Pothwar Plateau, Pakistan by analysing stomach contents. The research activities were conducted in major field crops including wheat-groundnut and in the fallow lands during non-crop season at the field boundaries. The specimens were captured from the fields using kill/snap traps, and dissected to collect their stomach samples for laboratory analysis. Light microscopic slides of the plant material were recovered from stomach samples and the reference materials were collected from the field. Results revealed that the bandicoot rat predominantly fed upon cultivated crops during cropping season but consumed wild vegetation during non-cropping season. There was no significance difference between summer and winter diets. Most frequently consumed crop food items were wheat (Triticum aestivum; 28.57%), groundnut (Arachis hypogea; 11.26%), sorghum (Sorghum bicolor; 10.17%), chickpea (Cicer arietinum; 9.52%), maize (Zea mays; 6.49%), millet (Pennisetum glaucum; 5.84%), barley (Hordeum vulgare; 4.98%) and mustard (Brassica campestris; 4.98%). Among wild vegetation were consumed khbal gha (Cynodon dactylon; 7.79%), baron dhab (Demostachya bipinnata; 7.36%) and Prickly flower (Achyranthes aspera; 3.03%). The study concludes that, in addition to consuming wheat and groundnut crops, the Lesser bandicoot rat also subsists on grasses, weeds, and some fodder crops, as important component of its diet in agro-ecosystem of the Pothwar Plateau.

Resumo Os hábitos alimentares de ratos bandicoot (Bandicota bengalensis) foram investigados nas plantações agrícolas do planalto de Pothwar, Paquistão, por meio da análise do conteúdo estomacal. As atividades da pesquisa foram conduzidas nas principais culturas de campo, incluindo trigo e amendoim, e em terras de pousio durante a estação não agrícola nos limites do campo. Os espécimes foram capturados dos campos usando armadilhas kill/snap e dissecados para coletar suas amostras de estômago para análise laboratorial. Lâminas de microscopia de luz do material vegetal foram recuperadas de amostras de estômago; os materiais de referência foram coletados no campo. Os resultados revelaram que o rato bandicoot alimentava-se predominantemente de culturas cultivadas durante a época de cultivo, mas consumia vegetação selvagem durante a época de não colheita. Não houve diferença significativa entre as dietas de verão e inverno. Os alimentos agrícolas mais frequentemente consumidos foram trigo (Triticum aestivum; 28,57%), amendoim (Arachis hypogea; 11,26%), sorgo (Sorghum bicolor; 10,17%), grão de bico (Cicer arietinum; 9,52%), milho (Zea mays; 6,49%), milheto (Pennisetum glaucum; 5,84%), cevada (Hordeum vulgare; 4,98%) e mostarda (Brassica campestris; 4,98%). Entre a vegetação silvestre foram consumidos khbal gha (Cynodon dactylon; 7,79%), barão dhab (Demostachya bipinnata; 7,36%) e flor espinhosa (Achyranthes aspera; 3,03%). O estudo conclui que, além de consumir culturas de trigo e amendoim, o rato bandicoot pequeno também subsiste de gramíneas, ervas daninhas e algumas culturas forrageiras, componentes importantes de sua dieta no agroecossistema do planalto de Pothwar.

Dietary habits of lesser bandicoot rat (Bandicota Bengalensis) in an agro-ecosystem, Pothwar Plateau, Pakistan / Hábitos alimentares de rato bandicoot (Bandicota Bengalensis) em um agroecossistema do Planalto de Pothwar, Paquistão

Abstract Dietary habits of bandicoot rats (bandicota bengalensis) were investigated in the agricultural crops of the Pothwar Plateau, Pakistan by analysing stomach contents. The research activities were conducted in major field crops including wheat-groundnut and in the fallow lands during non-crop season at the field boundaries. The specimens were captured from the fields using kill/snap traps, and dissected to collect their stomach samples for laboratory analysis. Light microscopic slides of the plant material were recovered from stomach samples and the reference materials were collected from the field. Results revealed that the bandicoot rat predominantly fed upon cultivated crops during cropping season but consumed wild vegetation during non-cropping season. There was no significance difference between summer and winter diets. Most frequently consumed crop food items were wheat (Triticum aestivum; 28.57%), groundnut (Arachis hypogea; 11.26%), sorghum (Sorghum bicolor; 10.17%), chickpea (Cicer arietinum; 9.52%), maize (Zea mays; 6.49%), millet (Pennisetum glaucum; 5.84%), barley (Hordeum vulgare; 4.98%) and mustard (Brassica campestris; 4.98%). Among wild vegetation were consumed khbal gha (Cynodon dactylon; 7.79%), baron dhab (Demostachya bipinnata; 7.36%) and Prickly flower (Achyranthes aspera; 3.03%). The study concludes that, in addition to consuming wheat and groundnut crops, the Lesser bandicoot rat also subsists on grasses, weeds, and some fodder crops, as important component of its diet in agro-ecosystem of the Pothwar Plateau.

Resumo Os hábitos alimentares de ratos bandicoot (Bandicota bengalensis) foram investigados nas plantações agrícolas do planalto de Pothwar, Paquistão, por meio da análise do conteúdo estomacal. As atividades da pesquisa foram conduzidas nas principais culturas de campo, incluindo trigo e amendoim, e em terras de pousio durante a estação não agrícola nos limites do campo. Os espécimes foram capturados dos campos usando armadilhas kill/snap e dissecados para coletar suas amostras de estômago para análise laboratorial. Lâminas de microscopia de luz do material vegetal foram recuperadas de amostras de estômago; os materiais de referência foram coletados no campo. Os resultados revelaram que o rato bandicoot alimentava-se predominantemente de culturas cultivadas durante a época de cultivo, mas consumia vegetação selvagem durante a época de não colheita. Não houve diferença significativa entre as dietas de verão e inverno. Os alimentos agrícolas mais frequentemente consumidos foram trigo (Triticum aestivum; 28,57%), amendoim (Arachis hypogea; 11,26%), sorgo (Sorghum bicolor; 10,17%), grão de bico (Cicer arietinum; 9,52%), milho (Zea mays; 6,49%), milheto (Pennisetum glaucum; 5,84%), cevada (Hordeum vulgare; 4,98%) e mostarda (Brassica campestris; 4,98%). Entre a vegetação silvestre foram consumidos khbal gha (Cynodon dactylon; 7,79%), barão dhab (Demostachya bipinnata; 7,36%) e flor espinhosa (Achyranthes aspera; 3,03%). O estudo conclui que, além de consumir culturas de trigo e amendoim, o rato bandicoot pequeno também subsiste de gramíneas, ervas daninhas e algumas culturas forrageiras, componentes importantes de sua dieta no agroecossistema do planalto de Pothwar.

Establishment of a rat model with aortic dissection induced by β-aminopropionitrile combined with angiotensin Ⅱ / 中国胸心血管外科临床杂志

@#Objective    To investigate the optimal administration combination of β-aminopropionitrile (BAPN) and Angiotensin Ⅱ (Ang-Ⅱ) in the establishment of SD rat aortic dissection (AD) model and the related complications. Methods    Forty-two three-week-old male SD rats were randomly divided into 7 groups: a group A (0.25% BAPN), a group B (0.40% BAPN), a group C (0.80% BAPN), a group D [1 g/(kg·d) BAPN], a group E [1 g/(kg·d) BAPN+ 1 μg/(kg·min) saline], a group F [1 g/(kg·d) BAPN+1 μg/(kg·min) Ang-Ⅱ] and a group G (control group). There were 6 rats in each group. The intervention period was 4 weeks (groups E and F were 4 weeks+5 days). Rats were dissected immediately if they died during the experiment. After the intervention, the surviving rats were sacrificed by pentobarbital sodium, and the whole aorta was separated and retained. Hematoxylin-eosin staining was used to observe the changes of aorta from the pathological morphology. Results    There was no statistical difference in the survival rate among the groups after 4 weeks of BAPN intervention (P>0.05). After 5 days of mini-osmotic pumps implantation, the survival rate of rats was higher in the group E than that in the group F (P=0.008), and the incidence of AD in the group E was lower than that in the group F (P=0.001). BAPN could affect the food and water intake of rats. After BAPN intervention for 4 weeks, the body weight of rats in the group G was higher than those in the intervention groups (P<0.05). BAPN combined with Ang-Ⅱ could make the aortic intima thick, elastic fiber breakage, arrangement disorder, and inflammatory cell infiltration in rats, which conformed to the pathological and morphological changes of AD. BAPN could also affect mental state and gastrointestinal tract. Conclusion    The combination of BAPN [1 g/(kg·d)] and Ang-Ⅱ [1 μg/(kg·min)] can stably establish AD model in rats, which will provide a stable carrier for further study of the pathogenesis and therapeutic targets of AD. However, the complications in this process are an unstable factor. How to balance the influence of BAPN on other tissues and organs in the process of AD model establishment remains to be further studied.

Prevention and treatment of lipoic acid-niacin on blue-light-induced retinal damage in rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the preventive effect and optimal drug dose of lipoic acid-niacin(N2L)against blue light-induced retinal damage in SD rats, and to explore its possible protective mechanism.METHODS: A total of 36 specific pathogen free-grade male SD rats of 150-200 g were selected and randomly divided into normal control group, blue light injury group, N2L low-dose group(1.0 mg/kg), N2L medium-dose group(2.5 mg/kg), N2L high-dose group(5.0 mg/kg), and physiological saline group, with 6 rats in each group. The normal control group was reared in a 12 h dark and light cycle, and the rest of the groups received 9 h of daily light exposure, 3 h of blue light irradiation with a wavelength of 455 nm and an intensity of 3000±50 lx, and 12 h of darkness to establish the injury model, and were exposed to light exposure for 14 d. For 14 consecutive durations, a 1 mL dose of the corresponding drug was injected intraperitoneally. The rats were reared for another 5 d with a regular 12 h light-dark cycle and were examined by electroretinography. Specimens were prepared by over anesthesia, HE staining, and the thickness of the outer nuclear layer was observed under a optical microscope; superoxide dismutases(SOD)activity was detected by CheKineTM SOD Activity Assay Kit; and the retinal Caspase-3, quinone oxidoreductase 1(NQO1), glutathione S transferase(GST), Bcl-2, and Bax protein expression in rat retina were detected by Western blot.RESULTS: The amplitude of b-wave in dark-vision ERG 3.0 and 10.0(cd·s)/m2 stimulated light, b-wave in bright-vision ERG 3.0(cd·s)/m2 stimulated light, and the amplitude of the 2nd wave peak of oscillatory potential were significantly lower in blue light injury group than that in the normal control group(all P&#x0026;#x003C;0.01), while the amplitude was significantly higher in the N2L medium-dose group than in the blue light injury group(all P&#x0026;#x003C;0.05), and was not statistically different from that of the normal control group; the thickness of the retina in the blue light injury group was decreased in the ONL compared with that of the normal control group(P&#x0026;#x003C;0.001), while in the N2L medium dose group, it was thicker than that of the blue light injury group(P&#x0026;#x003C;0.001), and there was no statistically significant difference from the normal control group; SOD activity was significantly higher in the N2L medium-dose group than in the remaining 5 groups(P&#x0026;#x003C;0.05); the expression of Caspase-3, Bax, and NQO1 in the blue light injury group was higher than that of the normal control group(all P&#x0026;#x003C;0.01), and expression of Bax and Caspase-3 was significantly lower in the N2L medium-dose group compared with the blue light injury group(all P&#x0026;#x003C;0.001), whereas GST, NQO1 and Bcl-2 were significantly increased(all P&#x0026;#x003C;0.01).CONCLUSION:A concentration of 2.5 mg/kg N2L can effectively antagonize the damaging effect of blue light on the retina of SD rats, and it is expected to be a preventive and curative drug for it.

Hedysari Radix Polysaccharide Regulates Apoptosis of Smooth Muscle Cells in Gastric Antrum of Rat Model of Diabetic Gastroparesis via IGF-1/PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effects of Hedysari Radix polysaccharide on the apoptosis of gastric sinus smooth muscle cells and explore the underlying mechanism via the insulin-like growth factor-1 （IGF-1）/phosphatidylinositol 3-kinase （PI3K）/serine-threonine kinase （Akt） pathway in the rat model of diabetic gastroparesis （DGP）. MethodSixty-two Wistar male rats were randomized into a blank group （n=12） and a modelling group （n=50）. The rat model of DGP was established by small-dose multiple intraperitoneal injections of streptozotocin combined with an irregular high-fat and high-sugar diet for 4 weeks. The modeled rats were randomized into model group， mosapride citrate （1.35 mg·kg-1）， and high-， medium-， and low-dose （200， 100， and 50 mg·kg-1， respectively） Hedysari Radix polysaccharide groups. The rats were administrated with corresponding drugs by gavage， and those in the blank and model groups with equal volumes of pure water by gavage once a day for 8 consecutive weeks. The random blood glucose and body mass were measured every 2 weeks， and gastric emptying rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of smooth muscle in gastric antrum， and terminal deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to detect the apoptosis of smooth muscle cells in the gastric antrum. The expression of IGF-1， phosphorylated （p）-PI3K， and p-Akt in the smooth muscle of gastric sinus tissue was detected by immunohistochemistry. Western blot was employed to determine the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the smooth muscle of the gastric antrum. ResultCompared with the blank group， the model group showed elevated random blood glucose at all time points （P<0.01）， decreased body mass and gastric emptying rate （P<0.01）， increased apoptotic index of smooth muscle cells in the gastric antrum （P<0.01）， down-regulated protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated protein level of Bax （P<0.01）. Compared with the model group， the 8 weeks of drug administration lowered the random blood glucose， increased the body mass and gastric emptying rate （P<0.05， P<0.01）， decreased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， up-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and down-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the mosapride citrate group，the administration of low-dose Hedysari Radix polysaccharide for 6 and 8 weeks lowered the random blood glucose and decreased the body mass （P<0.05， P<0.01），low and medium-dose Hedysari Radix polysaccharide decreased the gastric emptying rate and the apoptotic index of smooth muscle cells in the astragaloside low-dose group decreased （P<0.05）. The protein levels of IGF-1，p-PI3K/PI3K，p-Akt/Akt and Bcl-2（low dose）were down-regulated and the protein level of Bax was up-regulated by low doses of Hedysari Radix polysaccharide （P<0.05， P<0.01）. Compared with high-dose Hedysari Radix polysaccharide， low-dose Hedysari Radix polysaccharide elevated random blood glucose and reduced body mass after 6 and 8 weeks of administration （P<0.05， P<0.01）， and the low and medium doses decreased the gastric emptying rate， increased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， down-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the medium-dose group，the low-dose group of Hedysari Radix polysaccharide had lower body mass，lower gastric emptying rate in rats，higher apoptotic index of smooth muscle cells in gastric sinus tissue after 6 and 8 weeks of administration （P<0.05， P<0.01）， and lower protein expression of IGF-1，p-PI3K/PI3K，p-Akt/Akt. ConclusionHedysari Radix polysaccharide protects the smooth muscle cells in gastric antrum against apoptotic injury and promotes gastric motility by activating the IGF-1/PI3K/Akt signaling pathway， as manifested by the up-regulated expression of IGF-1， p-PI3K， p-Akt， and Bcl-2 and down-regulated expression of Bax.

Revisión histórica, distribución potencial y estado de conservación de la rata endémica de México Xenomys nelsoni (Rodentia: Cricetidae) / Historical review, potential distribution, and conservation of the Mexican endemic rat Xenomys nelsoni (Rodentia: Cricetidae)

Resumen Introducción: La Rata de Magdalena, Xenomys nelsoni, es un roedor endémico de México, de distribución restringida a las selvas bajas caducifolias densas, en una pequeña región de la costa del Pacífico mexicano. Es una especie poco conocida, catalogada como "En Peligro" de acuerdo con la Unión Internacional para la Conservación de la Naturaleza (IUCN). Este desconocimiento unido a la alta tasa de deforestación de su hábitat hace que su conservación sea prioritaria. Objetivo: Realizar un recuento histórico de los registros depositados en las colecciones científicas, generar mapas de distribución potencial y analizar el estado de conservación de la especie. Método: Los datos de ocurrencia de las especies se obtuvieron de la literatura y bases de datos digitales y se analizaron por décadas. Se utilizaron los programas GARP y MaxEnt para generar los modelos de nicho ecológico. La importancia de las variables en el modelo se estimó mediante un análisis Jackknife. Resultados: A lo largo de 129 años 19 recolectores registraron 69 ejemplares, de los cuales 65 están depositados en siete colecciones internacionales y una nacional. Aunque la especie sólo se ha recolectado en Jalisco y Colima, la distribución potencial de X. nelsoni incluye también el estado de Michoacán. De esta área estimada, sólo el 1.5 % se encuentra dentro de un Área Natural Protegida. Conclusiones: Los resultados de la distribución potencial podrían ser utilizados para verificar la presencia de la especie en lugares donde no ha sido recolectada como el norte de la Reserva de la Biosfera Chamela-Cuixmala y en algunas zonas de la provincia fisiográfica Costas del Sur en el estado de Michoacán. Es necesario incrementar los muestreos en regiones poco estudiadas predichas por el modelo y aumentar el área de protección.

Abstract Introduction: The Magdalena Rat, Xenomys nelsoni, is a rodent endemic to Mexico, whose distribution is restricted to dense tropical dry forests in a small region on the Pacific coast of Mexico. It is a poorly known species categorized as "Endangered" by the International Union for Conservation of Nature (IUCN). This lack of knowledge and the high rates of deforestation of its habitat makes its conservation a priority. Objective: To summarize the historical records deposited in scientific collections, to create potential distribution maps, and to analyze the conservation status of the species. Methods: We obtained species occurrence data from literature and digital databases, analyzing them by the decade. We used GARP and MaxEnt software to generate the ecological niche models. The importance of the variables in the model was estimated using the Jackknife technique. Results: Over 129 years, 19 collectors registered 69 specimens, of which 65 are deposited in one national and seven international collections. Although the species has only been collected in Jalisco and Colima, the potential distribution for X. nelsoni also includes the state of Michoacán. Of this estimated area, only 1.5 % is in a Protected Natural Area. Conclusions: The results of the potential distribution could be used to verify the presence of the species in places where it has not been collected, such as the northern part of the Chamela-Cuixmala Biosphere Reserve and in some areas of the physiographic province Costas del Sur in the state of Michoacán. It is needed to increase samplings in the least studied regions predicted by the model and expand the area of protection.

The Localization of R-Spondin1 and R-Spondin 3 Peptides in Rat Hypothalamus: An Immunohistochemical Study / Localización de los Péptidos R-Spondin1 y R-Spondin3 en el Hipotálamo de Rata: Un Estudio Inmunohistoquímico

SUMMARY: The R-spondin protein family is a group of proteins that enhance Wnt/b-catenin signaling and have pleiotropic functions in stem cell growth and development. In the literature reviews, there is no histomorphological study showing the localization and distribution of R-spondins in different hypothalamic nuclei. For this reason, the purpose of this study was to determine the localization, distribution characteristics, and densities in the hypothalamic nuclei of neurons expressing Rspo1 and Rspo3 proteins. The free-floating brain sections of the male rats who were not exposed to any treatment were stained with the indirect immunoperoxidase method using the relevant antibodies. As a result of the immunohistochemical studies, it was determined that neurons expressing the Rspo1 protein were found in large numbers in the supraoptic nucleus (SON), the suprachiasmatic nucleus (SCh), anterior paraventricular nucleus, periventricular hypothalamic nucleus (PeV), anterior hypothalamic area, magnocellular preoptic nucleus (MCPO) and the lateral hypothalamic area (LH) from the hypothalamic nuclei, while they were localized in fewer numbers in the arcuate nucleus (ARC). Rspo3 protein expression was found in neurons localized in the hypothalamic nuclei SON, paraventricular nucleus (PVN), PeV, ARC, ventromedial nucleus (VMH), LH, anterior parvicellular nucleus, and zona inserta (ZI). In addition, neurons synthesizing both peptides were found in the cortex and hippocampus regions (H). Rspo1 and 3 proteins are expressed in hypothalamic energy homeostatic areas, thus these proteins may be involved in the regulation of food intake.

La familia de proteínas R-espondina es un grupo de proteínas que mejoran la señalización de Wnt/b-catenina y tienen funciones pleiotrópicas en el crecimiento y desarrollo de las células madre. En las revisiones de la literatura no existen estudios histomorfológicos que muestren la localización y distribución de las R-espondinas en diferentes núcleos hipotalámicos. Por esta razón, el propósito de este estudio fue determinar la localización, características de distribución y densidades en los núcleos hipotalámicos de neuronas que expresan las proteínas Rspo1 y Rspo3. Secciones de cerebro flotantes de ratas macho que no fueron expuestas a ningún tratamiento se tiñeron con el método de inmunoperoxidasa indirecta utilizando los anticuerpos pertinentes. Como resultado de los estudios inmunohistoquímicos, se determinó que las neuronas que expresan la proteína Rspo1 se encontraron en gran número en el núcleo supraóptico (SON), el núcleo supraquiasmático (SCh), el núcleo paraventricular anterior, el núcleo hipotalámico periventricular (PeV), el núcleo hipotalámico anterior área, núcleo preóptico magnocelular (MCPO) y el área hipotalámica lateral (LH) de los núcleos hipotalámicos, mientras que se localizaron en menor número en el núcleo arqueado (ARC). La expresión de la proteína Rspo3 se encontró en neuronas localizadas en los núcleos hipotalámicos SON, núcleo paraventricular (PVN), PeV, ARC, núcleo ventromedial (VMH), LH, núcleo parvicelular anterior y zona inserta (ZI). Además, se encontraron neuronas que sintetizan ambos péptidos en las regiones de la corteza y el hipocampo (H). Las proteínas Rspo1 y 3 se expresan en áreas homeostáticas de energía hipotalámicas, por lo que estas proteínas pueden estar involucradas en la regulación de la ingesta de alimentos.

Anatomía Comparada del Bazo de la Rata Blanca (Rattus norvegicus albinus), Una Revisión de la Literatura / Comparative Anatomy of the Spleen of the White Rat, (Rattus norvegicus albinus) a Literature Review

El bazo es el órgano linfático intraperitoneal más grande del organismo, presentando dos funciones principales: defensiva, mediante respuesta inmunitaria y filtración sanguínea. El objetivo de la presente revisión, fue obtener información actualizada sobre la anatomía del bazo de la rata albina (Rattus norvegicus albinus) y comparativa con la anatomía del bazo humano, perro, gato y cerdo, al representar las principales especies de importancia en la medicina, medicina veterinaria y en las ciencias biomédicas. Se realizó una búsqueda de material bibliográfico actualizado en diferentes sitios web científicos. Es así como, se revisaron 71 fuentes bibliográficas, en su gran mayoría artículos científicos (31), libros de anatomía humana y veterinaria (17), artículos especializados (17) y tesis (6). En general existe consenso, sobre la descripción anatómica del bazo, el cual se sitúa en la región hipocondriaca izquierda del abdomen, entre el fondo del estómago y el diafragma, irrigado por la arteria y vena esplénica. Se evidenció que existen similitudes en aspectos macroscópicos, al comparar el bazo de la rata blanca, con el bazo de otras especies (funcionalidad, peso relativo, ubicación topográfica). En aspectos microscópicos, el bazo en humanos y otros mamíferos se compone de estroma, además de parénquima, constituido a su vez por pulpa blanca y roja. En particular, existen diferencias entre el bazo de rata, humano, gato, perro y cerdo, en formas, tamaños y aspectos microscópicos, relacionados con la microcirculación e inmunidad. Mientras que existen semejanzas en procesos patológicos y respuestas a tratamientos farmacológicos y clínicos. Por lo anteriormente expuesto, se concluye que la rata albina constituye un buen modelo biológico, específicamente en aspectos anatómicos microscópicos del bazo de tipo inmunológico. Mientras que el bazo de cerdo es mejor comparativamente, en estudios anatómicos macroscópicos de tipo quirúrgicos, resultando ambos extrapolables, especialmente a la medicina humana.

SUMMARY: The spleen is the largest intraperitoneal lymphatic organ of the body, presenting two main functions: defensive, through immune response and blood filtration. The objective of the present review was to obtain updated information on the anatomy of the spleen of the albino rat (Rattus norvegicus albinus) and to compare it with the anatomy of the human, dog, cat and pig spleen, representing the main species of importance in medicine, veterinary medicine and biomedical sciences. A search for updated bibliographic material was carried out in different scientific websites. Thus, 71 bibliographic sources were reviewed, mostly scientific articles (31), human and veterinary anatomy books (17), specialized articles (17) and theses (6). In general, there is consensus on the anatomical description of the spleen, which is located in the left hypochondriac region of the abdomen between the fundus of the stomach and the diaphragm, irrigated by the splenic artery and vein. It was evidenced that there are similarities in macroscopic aspects when comparing the spleen of the white rat with the spleen of other species (functionality, relative weight, topographic location). In microscopic aspects, the spleen in humans and other mammals is composed of stroma, in addition to parenchyma, constituted in turn by white and red pulp. In particular, there are differences between rat, human, cat, dog and pig spleens in shapes, sizes and microscopic aspects related to microcirculation and immunity. While there are similarities in pathological processes and responses to pharmacological and clinical treatments. For the above mentioned, it is concluded that the albino rat constitutes a good biological model, specifically in microscopic anatomical aspects of the spleen of immunological type. While the pig spleen is comparatively better in macroscopic anatomical studies of surgical type, both are extrapolable especially to human medicine.

The Effects of White Tea (Camellia Sinensis) and Infliximab Against Cisplatin- Induced Testicular Damage via Oxidative Stress and Apoptosis / Efectos del Té Blanco (Camellia Sinensis) e Infliximab Contra el Daño Testicular Inducido por Cisplatino a Través del Estrés Oxidativo y la Apoptosis

SUMMARY: Cisplatin (Cis) is an important chemotherapeutic agent used in cancer treatment. Males exposed to Cis were reported to exhibit testicular toxicity. Cis-induced testicular toxicity is mediated by oxidative stress, inflammation, testosterone inhibition and apoptosis. Accordingly, this study was conducted to evaluate the potential protective roles of infliximab (IFX), which is an anti- TNF-a agent, and of white tea (Camellia sinensis), which is known to possess antioxidant, anti-apoptotic, and anti-inflammatory effects, against Cis-induced testicular toxicity in rats. Rats were randomly assigned into five groups as follows: control group, Cisplatin (7 mg/kg) treatment group, Cisplatin (7 mg/kg) + infliximab (7 mg/kg) treatment group, cisplatin + white tea (WT) treatment group, and Cisplatin+ WT+IFX combined treatment group. In the present study, Cis exposure reduced the sperm count. It also increased testicular oxidative stress as well as the levels of inflammatory and apoptotic markers. Histopathological assays supported the biochemical findings. Treatment with IFX and/or WT restored testicular histology, preserved spermatogenesis, suppressed oxidative stress and apoptosis, and significantly ameliorated Cis-induced damage. It was concluded that white tea and infliximab could potentially serve as therapeutic options for the protection of testicular tissue against the harmful effects of Cis.

El cisplatino (Cis) es un importante agente quimioterapéutico utilizado en el tratamiento del cáncer. Se informó que los hombres expuestos a Cis exhibieron toxicidad testicular. La toxicidad testicular inducida por Cis está mediada por el estrés oxidativo, la inflamación, la inhibición de la testosterona y la apoptosis. En consecuencia, este estudio se realizó para evaluar las posibles funciones protectoras de infliximab (IFX), un agente anti-TNF-α, y del té blanco (Camellia sinensis), conocido por sus propiedades antioxidantes, antiapoptóticas y anti-TNF-α -efectos inflamatorios, contra la toxicidad testicular inducida por Cis en ratas. Cinco grupos de ratas se asignaron al azar de la siguiente manera: grupo control, grupo de tratamiento con cisplatino (7 mg/ kg), grupo de tratamiento con cisplatino (7 mg/kg) + infliximab (7 mg/kg), grupo de tratamiento con cisplatino + té blanco (WT), y grupo de tratamiento combinado Cisplatino+ WT+IFX. En el presente estudio, la exposición a Cis redujo el conteo de espermatozoides. También aumentó el estrés oxidativo testicular, así como los niveles de marcadores inflamatorios y apoptóticos. Los ensayos histopatológicos respaldaron los hallazgos bioquímicos. El tratamiento con IFX y/o WT restauró la histología testicular, preservó la espermatogénesis, suprimió el estrés oxidativo y la apoptosis, y mejoró significativamente el daño inducido por Cis. Se concluyó que el té blanco y el infliximab podrían potencialmente servir como opciones terapéuticas para la protección del tejido testicular contra los efectos nocivos de Cis.

Altered Chief Cell Morphology in the Gastric Gland of Streptozotocin-Diabetic Rats

SUMMARY: Diabetes mellitus (DM) is a metabolic disorder with rising incidences worldwide. Gastric symptoms of DM have been reported, including nausea, vomiting, bloating, and epigastric pain. Moreover, acute to chronic gastritis and atrophic gastritis occur in DM can affect the chief cells of the gastric gland. Chief cells are vital because of their ability to digest and separate vitamin B12 from protein. Lack of vitamin B12 leads to impaired DNA synthesis and abnormal metabolism in red blood cells, and eventually leading to pernicious anemia. Furthermore, decreased vibratory and positional senses, numbness, ataxia with subacute combined degeneration, and dementia are present in pernicious anemic patients. Twenty-four male adult Sprague-Dawley rats were used in this study. The rats were divided into control (n = 12) and diabetic (n = 12) groups. The rats were further separated into two categories: short-term (4 weeks) and long-term (24 weeks) groups. DM model was induced by manually injecting intraperitoneally with streptozotocin in citrate buffer at a dose of 60 mg/kg body weight. The same amount of buffer was injected into the control group. After sacrifice, three regions of the stomach (the cardia, body, and pylorus) were dissected. Histopathology was performed by staining with toluidine blue. Image analysis was used to quantify the zymogen granule accumulation in chief cells. The data were compared between the control and DM rats in each period using Student's t-test. In addition, transmission electron microscopy (TEM) was also used to examine the ultrastructures. There was a significant decrease in the percentage of zymogen granules in DM rats. Under TEM, the destructions of mitochondria, rough endoplasmic reticulum, and Golgi apparatus in the DM rat were observed in the chief cells. In rats with uncontrolled diabetes, there is damage to the chief cells all over the area of the stomach, affecting digestion and malabsorption of vitamin B12. Therefore, this result helps clinicians recognize that diabetic patients with gastric symptoms may have hidden pernicious anemia.

La diabetes mellitus (DM) es un trastorno metabólico con incidencia creciente a nivel mundial. Se han informado síntomas gástricos de DM, que incluyen náuseas, vómitos, distensión abdominal y dolor epigástrico. Además, la gastritis aguda a crónica y la gastritis atrófica que ocurren en la DM pueden afectar las células principales de la glándula gástrica. Las células principales son vitales debido a su capacidad para digerir y separar la vitamina B12 de las proteínas. La falta de vitamina B12 conduce a una síntesis de ADN deteriorada y un metabolismo anormal en los glóbulos rojos, lo que eventualmente conduce a una anemia perniciosa. Además, los pacientes con anemia perniciosa presentan disminución de los sentidos vibratorio y posicional, entumecimiento, ataxia con degeneración combinada subaguda y demencia. En este estudio se usaron 24 ratas Sprague-Dawley macho adultas. Las ratas se dividieron en grupos control (n = 12) y diabéticas (n = 12). Las ratas se separaron además en dos categorías: grupos a corto plazo (4 semanas) y a largo plazo (24 semanas). El modelo de DM se indujo inyectando manualmente por vía intraperitoneal estreptozotocina en tampón de citrato a una dosis de 60 mg/kg de peso corporal. Se inyectó la misma cantidad de tampón en el grupo control. Después del sacrificio, se disecaron tres regiones del estómago (cardias, cuerpo y píloro). La histopatología se realizó mediante tinción con azul de toluidina. El análisis de imágenes se utilizó para cuantificar la acumulación de gránulos de zimógeno en las células principales. Los datos se compararon entre las ratas control y DM en cada período utilizando la prueba t de Student. Además, se utilizó microscopía electrónica de transmisión (TEM) para examinar la ultraestructura celular. Hubo una disminución significativa en el porcentaje de gránulos de zimógeno en ratas DM. Bajo TEM, se observaron en las células principales las destrucción de las mitocondrias, del retículo endoplásmico rugoso y del complejo golgiense en la rata DM. En ratas con diabetes no controlada, hay daño en las células principales de toda el área del estómago, lo que afecta la digestión y la malabsorción de vitamina B12. Por lo tanto, este resultado ayuda a los médicos a reconocer que los pacientes diabéticos con síntomas gástricos pueden tener una anemia perniciosa oculta.

Localization of Diacylglycerol Kinase ζ in the Kidney of Adult Rats: Its Dominant Expression in Collecting Tubules and its Selective Lack in Proximal Tubules / Localización de Diacilglicerol Quinasa ζ en el Riñón de Ratas Adultas: su Expresión Dominante en Túbulos Colectores y su Carencia Selectiva en Túbulos Proximales

SUMMARY: Diacylglycerol kinase (DGK) exerts balancing the intracellular level between two-second messengers, diacylglycerol and phosphatidic acid, by its phosphorylation activity. DGK ζ is often localized in cell nuclei, suggesting its involvement in the regulation of intranuclear activities, including mitosis and apoptosis. The present immunohistochemical study of rat kidneys first revealed no detection levels of DGK ζ -immunoreactivity in nuclei of most proximal tubule epithelia in contrast to its distinct occurrence in cell nuclei of collecting and distal tubules with the former more dominant. This finding suggests that DGK ζ is a key factor regulating vulnerability to acute kidney injury in various renal tubules: its low expression represents the high vulnerability of proximal tubule cells, and its distinct expression does the resistance of collecting and distal tubule cells. In addition, this isozyme was more or less localized in nuclei of cells forming glomeruli as well as in endothelial nuclei of peritubular capillaries and other intrarenal blood vessels, and epithelial nuclei of glomerular capsules (Bowman's capsules) and renal calyces, including intrarenal interstitial cells.

La diacilglicerol quinasa (DGK) ejerce el equilibrio del nivel intracelular entre dos segundos mensajeros, diacilglicerol y ácido fosfatídico, por su actividad de fosforilación. La DGK ζ a menudo se localiza en los núcleos celulares, lo que sugiere su participación en la regulación de las actividades intranucleares, incluidas la mitosis y la apoptosis. El presente estudio inmunohistoquímico en riñones de rata no reveló niveles de detección de inmunorreactividad de DGK ζ en los núcleos de la mayoría de los epitelios de los túbulos proximales, en contraste a la detección en los núcleos celulares de los túbulos colectores y distales, siendo el primero más dominante. Este hallazgo sugiere que DGK ζ es un factor clave que regula la vulnerabilidad a la lesión renal aguda en varios túbulos renales: su baja expresión representa la alta vulnerabilidad de las células del túbulo proximal, y su expresión distinta hace a la resistencia de las células del túbulo colector y distal. Además, esta isoenzima estaba más o menos localizada en los núcleos de las células que forman los glomérulos, así como en los núcleos endoteliales de los capilares peritubulares y otros vasos sanguíneos intrarrenales, y en los núcleos epiteliales de las cápsulas glomerulares (cápsulas de Bowman) y los cálices renales, incluidas las células intersticiales intrarrenales.

Captopril inhibits thioacetamide-induced chronic liver injury associated with the suppression of inflammation / hypoxia- inducible factor 1-alpha / profibrogenic axis-mediated hepatotoxicity in rats / Captopril inhibe la lesión hepática crónica inducida por tioacetamida asociada con la supresión de la inflamación/factor 1-alfa inducible por hipoxia/hepatotoxicidad mediada por el eje profibrogénico en rata

SUMMARY: Liver transplantation is the only available method to treat liver failure induced by chronic liver injury. We sought to determine whether the angiotensin-converting enzyme inhibitor, captopril, can inhibit the development of chronic liver injury induced by the hepatotoxic agent thioacetamide (TAA) in association with the suppression of inflammation (hsCRP, TNF-α, and IL-6) / hypoxia- inducible factor 1-alpha (HIF-1α) / profibrosis (TIMP-1, MMP-9, and α-SMA) axis that mediates liver injury. Therefore, the model group of rats was injected for eight weeks with 200 mg/kg TAA starting at week two. The protective group was pretreated with 150 mg/ kg captopril daily for two weeks prior to TAA injections and continued receiving both capropril and TAA agents until being humanely scrificed at week 10. We observed a substantial damage to liver tissue in the model group as demonstrated by a significant (p<0.0001) increase in blood and hepatic tissue levels of high sensitivity C-reactive protein (hsCRP), tumor necrosis factor-a (TNF-α), interleukin- 6 (L-6), HIF-1α, tissue inhibitor of metalloproteinases-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), alpha-smooth muscle actin (α-SMA), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). All these parameters were significantly (p<0.0244) protected by captopril. Also, a significant (p<0.0001) positive correlation was observed between a-SMA (profibrosis) and the serum and tissue levels of hsCRP, TNF-α, HIF-1α, TIMP-1, MMP-9, and ALT. Thus, these findings suggest that the induction of chronic liver injury by the hepatotoxic compound, TAA is associated with the upregulation of inflammation/HIF-1α/profibrosis, with captopril exhibiting beneficial hepatic pleotropic effects.

El trasplante de hígado es el único método disponible para tratar la insuficiencia hepática inducida por una lesión hepática crónica. Buscamos determinar si el inhibidor de la enzima convertidora de angiotensina, captopril, puede inhibir el desarrollo de lesión hepática crónica inducida por el agente hepatotóxico tioacetamida (TAA) en asociación con la supresión de la inflamación (hsCRP, TNF-α e IL-6) / factor inducible por hipoxia 1-alfa (HIF-1α) / profibrosis (TIMP-1, MMP-9 y α- SMA) eje que media la lesión hepática. Por lo tanto, al grupo modelo de ratas se le inyectó durante ocho semanas 200 mg/kg de TAA a partir de la semana dos. El grupo protector fue pretratado con 150 mg/kg de captopril al día durante dos semanas antes de las inyecciones de TAA y continuó recibiendo capropril y agentes TAA hasta que fue sacrificado en la semana 10. Observamos un daño sustancial en el tejido hepático en el grupo modelo, como lo demuestra un aumento significativo (p<0,0001) de los niveles en sangre y tejido hepático de proteína C reactiva de alta sensibilidad (hsCRP), factor de necrosis tumoral-α (TNF-a), interleucina-6 (L-6), HIF-1α, inhibidor tisular de metaloproteinasas-1 (TIMP-1), metaloproteinasa de matriz-9 (MMP-9), actina de músculo liso alfa (α-SMA), alanina aminotransferasa (ALT) y aspartato aminotransferasa (AST). Todos estos parámetros estaban significativamente (p<0,0244) protegidos por captopril. Además, se observó una correlación positiva significativa (p<0,0001) entre α-SMA (profibrosis) y los niveles séricos y tisulares de hsCRP, TNF-α, HIF-1α, TIMP- 1, MMP-9 y ALT. Por lo tanto, estos hallazgos sugieren que la inducción de daño hepático crónico por el compuesto hepatotóxico, TAA, está asociada con la regulación al alza de la inflamación/HIF-1α/profibrosis, con captopril exhibiendo efectos pleotrópicos hepáticos beneficiosos.

SARS-CoV-2 antigen detection in deceased bodies: implications for infection prevention

Background & objectives: High transmissibility of the SARS-CoV-2 has significant implications on healthcare workers’ safety, preservation, handling, transportation and disposal of the deceased bodies. The objective of this study was to detect SARS-CoV-2 antigen in nasopharyngeal samples and its implications in handling and care of COVID-19 deceased bodies. Methods: A study was conducted at a dedicated COVID-19 centre on deceased individuals from April to December 2020. Rapid antigen test (RAT) and reverse transcription (RT)-PCR was compared on all the SARS-CoV-2 positive cadavers recruited in the study. Results: A total of 115 deceased individuals were included in the study. Of these, 79 (68.7%) were male and 36 (31.3%) were female and majority were in the age group of 51-60 yr [31 (27%)]. SARS-CoV-2 antigen test was positive in 32 (27.8%) and negative in 83 (72.1%) individuals. The mean time interval between deaths to the sample collection was 13.2 h with interquartile range of eight to 20 h. Reverse transcription (RT)-PCR was used as the reference test and 24 (20.9%) cases were true positive; 93.6 per cent [95% confidence interval (CI) 88.8-98.4%] sensitivity, 45.2 per cent (95% CI 35.5-55%) specificity, 60.2 per cent (95% CI 50.6-69.8%) positive predictive value and 88.8 per cent (95% CI 82.7-95%) negative predictive value of antigen test was computed. Interpretation & conclusions: SARS-CoV-2 antigen test was positive beyond 19 h in COVID-19 deceased individuals. Antigen test was found to be highly sensitive in the deceased. Patients, suspected of having died due to COVID-19, can be screened by this method. As infectiousness of the virus in the deceased bodies cannot be directly concluded from either the antigen or RT-PCR test, yet possible transmission cannot be completely ruled out. Strict infection control measures need to be followed during the handling and clearance of COVID-19 cadavers.

Effects of simulated gas of warm pressure charge explosion on neurobehavior and hippocampal neurogenesis in rats / 中华行为医学与脑科学杂志

Objective:To investigate the effects of thermobaric charge explosion simulated gas on long-term neurobehavior and hippocampal neurogenesis in rats.Methods:A total of 48 male SPF grade SD rats aged 8-10 weeks were randomly divided into control group, 5 min exposure group, 10 min exposure group and 15 min exposure group, with 12 rats in each group. Twenty-eight days after inhalation of infection, the anxiety-like behavior of rats was evaluated by an elevated cross maze, and the learning and memory function of rats was evaluated by two-way active avoidance experiment. The number of positive cells of rat hippocampal dentate gyrus neural stem cells marker molecule neural epithelial cell protein (SOX2) and mature neuron marker molecular neuronal nuclei (NeuN) was detected by immunofluorescence staining. Western blot was used to detect SOX2 and NeuN protein expression in the hippocampal tissues of rats. GraphPad prism 8.0 software was used for data analysis.The comparison of repeated measurement design data was carried out by repeated measurement ANOVA.One-way ANOVA was used for inter group comparisons, and Tukey test was used for pairwise comparison. Hippocampal nerve cells were counted using the Image J software.Results:(1) The experimental results of the elevated cross maze showed that the percentage of arm opening and the percentage of open arm residence time in each group had significant group effects ( F=22.31, 5.43, all P<0.05). The percentage of open arm entry times of rats in the 5 min, 10 min and 15 min exposure group ((28.85±1.47)%, (15.04±4.69)%, (12.66±2.89)%) and the percentage of residence time in open arm ((12.12±2.64)%, (12.16±1.11)%, (8.73±3.52)%) were all lower than those of the control group ((65.40±1.86)%, (42.92±3.12)%) (all P<0.05). There were no statistically significant differences in pairwise comparison among the three exposure groups (all P>0.05). (2)During the memory acquisition period, the results of repeated-ANOVA showed that the time main effect ( F=56.46), the group main effect ( F=16.64) and the interaction effect had significant differences( F=4.21)(all P<0. 05). The difference values of active avoidance number between the 4th day and 1st day among the four groups were significant different ( F=68.63, P<0.05). During the memory reproduction period, there were significant differences in active avoidance number and active avoidance time among the four groups ( F=8.17, 8.28, both P<0.05). The active avoidance numbers in 10 min and 15 min exposure groups((2.50±0.26) times, (2.33±0.06) times)were significantly lower than those in the control group ((8.33±3.72) times) (both P<0.05), and the active avoidance time ((6.25±0.40)s, (6.61±1.63)s) were significantly higher than those in the control group((3.69±1.41)s) (both P<0.05). The active avoidance numbers in 10 min and 15 min exposure groups were significantly lower than that in 5 min exposure group (both P<0.05). (3) The results of immunofluorescence staining showed that the numbers of SOX2-positive cells in the four groups were statistically significant ( F=5.33, P<0.05). The SOX2-positive cells in 15 min exposure group (4.33±1.12) was significantly lower than that in control group (7.67±1.52) ( P<0.05). The numbers of NeuN-positive cells in the four groups were significantly different ( F=11.06, P<0.05), and the NeuN-positive cells in the 10 min and 15 min exposure groups((105.67±8.50), (88.33±9.50)) were significantly lower than that in the control group (127.00±6.56) ( P<0.05). The NeuN-positive cells in 15 min exposure group were significantly lower than that in 5 min exposure group (110.67±8.32) ( P<0.05). (4) Western blot results showed that the relative expression of SOX2 and NeuN proteins in the four groups was statistically significant ( F=11.560, 7.035, both P<0.05). The relative expression of SOX2 and NeuN proteins in the 15 min exposure group were significantly lower than those in control group (both P<0.05). The relative expression of SOX2 protein in 15 min exposure group was significantly lower than that in 5 min exposure group ( P<0.05). Conclusion:Acute exposure to warm pressure charge explosion simulated gas can lead to anxiety-like behavior, learning and memory deficits in rats, and significantly reduce the protein expression levels of hippocampal dentate gyrus neural stem cells and mature neuronal marker molecules SOX2 and NeuN.

Effects of Erastin on depressive- and anxiety-like behavior and the expression of hippocampal ferroptosis-related proteins in rats / 中华行为医学与脑科学杂志

Objective:To explore the effects of intrahippocampal injection of ferroptosis inducer Erastin on depressive- and anxiety-like behavior and the expression of ferroptosis-related proteins in rats.Methods:Forty 6-week-old healthy male Sprague-Dawley rats were randomly divided into five groups ( n=8/group): Control group, Erastin low-dose(200 ng/μL) group, Erastin medium-dose(400 ng/μL) group, Erastin high-dose group(600 ng/μL) and lipopolysaccharide (LPS, 10 μg/L) group.After the intrahippocampal injection of Erastin(2.5 μL per side), body weight, and behavioral tests, including sucrose preference test (SPT), forced swimming test (FST), open field test (OFT), and elevated plus maze (EPM), were performed to evaluate depressive- and anxiety-like phenotypes from the fourth day after injection.The levels of ferroptosis-related proteins and mRNA, including glutathione peroxidase 4 (GPX4), cyclo-oxygenase 2 (COX2), ferritin heavy polypeptide 1 (FTH1), long-chain fatty acyl-CoA synthetase 4 (ACSL4), solute carrier family 7 member 11 (SLC7A11) were measured using real-time quantitative PCR and Western blot analysis.SPSS 22.0 software was used for statistical analysis.One-Way ANOVA was used for multi-group comparison, and LSD was used for further pound-wise comparison. Results:(1)Body weight and behavioral tests: there were no statistically significant differences in baseline body weight and behavioral tests in these groups ( F=0.02-1.15, all P>0.05). After intrahippocampal injection, compared with the control group, medium-dose Erastin induced depression-like behaviors in rats more significantly, as indicated by reduced bodyweight ((245.20±5.24)g, (267.45±13.16)), sucrose preference in SPT ((32.14±8.51)%, (68.17±13.67)%), central time in OFT ((6.01±2.57)s, (16.49±7.21)s), percentage of time in open arm in EPM ((5.00±3.83)%, (19.63±5.91)%) and increased immobility time in FST ((37.00±7.58)s, (12.50±5.51)s) and percentage of time in closed arm in EPM ((89.43±4.77)%, (59.96±9.91)%), and there were statistically significant differences in these groups (all P<0.05). (2)The expression of ferroptosis-related indicators: after intrahippocampal injection, the expression of mRNA ( F=2.23, 8.37, 2.91, 7.60, 3.16, all P<0.05) and protein ( F=3.31, 40.13, 8.52, 3.70, 70.79, all P<0.05) of FTH1, GPX4, SLC7A11, COX2 and ACSL4 in hippocampus were statistically significant differences in the 5 groups.The mRNA and protein levels of FTH1, GPX4 and SLC7A11 in Erastin medium-dose group were lower than those in the control group (all P<0.05), while the mRNA and protein levels of COX2 and ACSL4 were higher than those in the control group (all P<0.05). Conclusion:Intrahippocampal microinjection of Erastin(400 ng/μL) can induce ferroptosis in hippocampus of rats and can also induce depressive-like behaviors in rats.

Effect of Bushen Jianpi Kaixin formula on cognitive function and the expression of autophagy related proteins in Alzheimer’s disease model rats / 中华行为医学与脑科学杂志

Objective:To investigate the effect and mechanism of the Bushen Jianpi Kaixin formula on the learning and memory ability of Alzheimer's disease (AD) model rats.Method:A total of 72 SPF grade male SD rats were divided into control group, model group, Bushen group, Jianpi group, Kaixin group and Bushen Jianpi Kaixin group according to the random number table method ( n=12 in each group). The rats were intraperitoneally injected with D-galactose once a day for 6 weeks to replicate the model of AD.And the rats in different medication groups were given corresponding administration (Bushen formula: gavage 3.60 g·kg -1·d -1, Jianpi formula: gavage 4.05 g·kg -1·d -1, Kaixin formula: gavage 2.34 g·kg -1·d -1, Bushen Jianpi Kaixin formula: gavage 9.99 g·kg -1·d -1), while rats in control group and model group were treated with equal volume of 0.9% sodium chloride solution once a day for 28 days.The learning and memory ability was tested by Morris water maze.The expressions of phosphoinositide 3-kinase(PI3K), protein kinase B(Akt) and mammalian target of rapamycin(mTOR) in cerebral cortical tissues were detected by immunohistochemistry.The relative mRNA levels of p62 and Beclin in brain cortical tissue were detected by RT-PCR.SPSS 25.0 software was used for data processing, one-way ANOVA was used for inter group comparisons, and LSD test was used for further pairwise comparisons. Results:Morris water maze results showed statistically significant differences in escape latency and the times of crossing platform among the six groups ( F=368.10, 47.43, both P<0.01). The escape latency of Bushen Jianpi Kaixin group((29.30±1.64) s) was shorter than that of model group((55.58±3.23) s) ( P<0.01), the times of crossing platform ((5.17±0.72) times) in Bushen Jianpi Kaixin group was higher than that of model group (1.50±0.52)time, P<0.01). Compared with the Bushen Jianpi Kaixin group, the escape latencies of Bushen group, Jianpi group and Kaixin group were longer (all P<0.01), the times of crossing platform in Bushen group was lower ( P<0.01). Immunohistochemical results showed statistically significant differences in the positive protein expression of PI3K, Akt, and mTOR proteins among the six groups ( F=68.52, 22.22, 31.52, all P<0.01). Compared with the model group, the levels of positive protein of PI3K ((0.47±0.15), (0.57±0.12)), Akt ((0.31±0.02), (0.38±0.02)), and mTOR ((0.22±0.18), (0.28±0.11)) in Bushen Jianpi Kaixin group were less (all P<0.01). Compared with the Bushen Jianpi Kaixin group, the levels of positive protein of PI3K and mTOR of Bushen group, Jianpi group and Kaixin group were higher (all P<0.01). RT-PCR results showed statistically significant differences in the relative mRNA levels of Beclin and p62 among all the groups ( F=8.79, 21.01, both P<0.01). The relative mRNA level of Beclin in Bushen Jianpi Kaixin group was higher than that of the model group ((0.97±0.07), (0.64±0.12)), and the relative mRNA level of p62 of Bushen Jianpi Kaixin group was less than that of model group((0.98±0.16), (1.16±0.24))(both P<0.01). The relative mRNA levels of p62 in Bushen group, Jianpi group and Kaixin group were higer than those of Bushen Jianpi Kaixin group (all P<0.05). Conclusion:Bushen Jianpi Kaixin formula can improve cognitive impairment and learning and memory ability in AD model rats.The mechanism may be related to the regulation of PI3K/Akt/mTOR autophagy pathway.The combination prescription is better than the split prescription.

Mechanism of anterior cingulate cortex in modulating pain and anxiety-like behavior in chronic non-specific low back pain model rats / 中华行为医学与脑科学杂志

Objective:To explore the potential mechanisms of anterior cingulate cortex (ACC) in modulating pain behavior and anxiety-like behavior of rats with chronic non-specific low back pain induced by nerve growth factor (NGF).Methods:Ninety-six male SPF grade SD rats aged 8 weeks were randomly divided into four groups according the random number table method: control group, model group, control+ D-2-amino-5-phosphonopentanoate (D-AP5) group (control+ D-AP5 group) and model+ D-AP5 group, with 24 rats in each group.Low back pain model of rat was established by injection of NGF into multifidus muscle (left side) of the low backs of rats(two times with a five-day interval). Five days after modeling, rats in model+ D-AP5 group and control+ D-AP5 group were injected with the N-methyl-D-aspartate (NMDA) receptor antagonist D-AP5(2 μg, 0.3 μL) at the right side of the ACC once a day for consecutive 3 days, and rats in control group and model group were injected with the same amount of 0.9% sodium chloride solution. Seven days after modeling, the pain threshold of rats was evaluated by mechanical stimulation test and hot and cold plate test.The anxiety-like behavior was tested by open field test.The density of glial fibrillary acidic protein (GFAP) positive cells and c-Fos(a kind of immediate early gene) positive cells of the spinal cord were observed by immunofluorescence. The expression of GFAP, c-Fos, phosphorylated-c-Jun N-terminal kinases (p-JNK), monocyte chemoattractant protein-1 (MCP-1), and chemokine (C-X-C motif) ligand 1 (CXCL-1) proteins in the L2 segment of the spinal cord were detected by Western blot. SPSS 23.0 software was used for statistical analysis. One-way ANOVA was used to analyze normal distribution measurement data for comparison among multiple groups, and Tukey test was used for further pairwise comparisons. The Kruakal-Wallis H test was used for non-normal distribution measurement data, and Mann-Whitney U test was used for further pairwise comparisons with Bonferroni-corrected P-values. Results:In the experiments measuring pressure pain threshold (PPT) and paw withdrawal threshold (PWT), there were statistically significant differences in the PPT and PWT of rats among the four groups ( F=53.498, 41.939, both P<0.001). Seven days after modeling, PPT ((418.5±46.9) g) and PWT ( (55.6±7.1) g) in the ipsilateral side of the rats in model+ D-AP5 group were higher than those in model group ((290.0±32.0) g, (30.5±7.5) g) (both P<0.001). In the open field test, there were statistically significant differences in percentage of the inner zone distance ( H=11.922, P<0.01) and the percentage of inner zone time ( H=21.614, P<0.001) of rats among the four groups. The percentage of inner zone time in model+ D-AP5 group was higher than that in model group (5.6(4.3, 7.9) %, 3.1(2.1, 3.8) %) ( P<0.01). The results of immunofluorescence showed that there were statistically significant differences in the density of GFAP positive cells and c-Fos positive cells at the ipsilateral side of the superficial laminae of rats among the four groups ( H=49.085, F=18.120, both P<0.001). The density of GFAP positive cells (34.3(21.1, 47.5) cells/mm 2) and c-Fos positive cells ((52.7±39.4) cells/mm 2) at the ipsilateral side of the superficial laminae in model+ D-AP5 group were less than those in model group (76.5(68.6, 94.9) cells/mm 2, (112.4±63.7) cells/mm 2) (both P<0.001). The Western blot results showed that there were statistically significant differences in the protein expression of GFAP, c-Fos, p-JNK, MCP-1 and CXCL-1 in the L2 segment of rats among the four groups ( F=49.413, 38.437, 41.867, 36.735, 130.951, all P<0.001). The protein expression of GFAP (1.7±0.5), c-Fos (1.1±0.1), p-JNK (1.7±0.3), MCP-1 (1.0±0.4) and CXCL-1 (0.8±0.1) in the L2 segment in model+ D-AP5 group were lower than those in model group ((4.3±0.7), (2.6±0.5), (2.8±0.4), (2.9±0.4), (3.5±0.4)) (all P<0.01). Conclusion:ACC modulates mechanical hyperalgesia and anxiety-like behavior in chronic non-specific low back pain rats, which might be associated with the involvement of spinal astrocytes, p-JNK signal pathway and chemokines such as MCP-1 and CXCL-1.

Protective effects and mechanisms of L-carnitine on cognitive dysfunction in rats with chronic cerebral hypoperfusion / 中华行为医学与脑科学杂志

Objective:To explore the protective effects and mechanisms of L-carnitine (LCAR) on cognitive dysfunction in chronic cerebral hypoperfusion rats.Methods:Totally 90 SD male rats (SPF class) aged 3-4 months were divided into four groups according to random number talbe: sham operated control group (SHAM group, n=15), sham operated with L-carnitine treatment group (LCAR group, n=25), 2-vessel occlusion group (2VO group, n=25), and 2-vessel occlusion with L-carnitine treatment group (2VO+ LCAR group, n=25). The chronic cerebral hypoperfusion model was established by bilateral common carotid artery ligation, and the carotid arteries from SHAM group and LCAR group were only separated without ligation.L-carnitine was administered intraperitoneally (300 mg·kg -1·d -1) for 30 days after surgery in the LCAR and 2VO+ LCAR groups.After 30 days of L-carnitine intervention, Morris water maze was performed to test the spatial cognitive function of the rats, the ATP level of hippocampal tissue was detected by chemiluminescence, the mitochondrial structure and synaptic structure of hippocampal neurons were observed by transmission electron microscopy, the degree of mitochondrial damage was scored, the vesicle density was counted and measured, the level of N-methyl-D-aspartate receptor subunit 2A or 2B(NR2A/B) and postsynaptic density 95(PSD95) in hippocampal tissue were detected by Western blot.The expression and distribution levels of transcription factor cAMP response element-binding protein(CREB) in brain tissues were observed by immunofluorescence.SPSS 16.0 software was used for statistical analysis.The escape latency data of repeated learning training in Morris water maze was conducted by repetitive measurement ANOVA, while other data were adopted by one-way ANOVA, and Dunnett's t test was used for further pairwise comparison. Results:(1)Morris water maze results showed that the time and group interaction of escape latency was not significant among the 4 groups of rats ( F=1.4, P>0.05), but the time main effect and group main effect were significant( F=21.6, 15.2, both P<0.05). Morris water maze results showed that platform position learning from 3rd to 7th day, the escape latencies in 2VO group were longer than those in SHAM group and 2VO+ LCAR group (all P<0.05). The results of short-term memory showed that the escape latency in 2VO group was longer than those in SHAM group and 2VO+ LCAR group (all P<0.05). Meanwhile, the retention time and crossing times in the platform area of 2VO group were less than those in SHAM group and 2VO+ LCAR group (all P<0.05). (2) The absolute and relative levels of ATP in hippocampus showed that the difference among the 4 groups were statistically significant ( F=14.6, 13.2, both P<0.05). ATP level of hippocampus in 2VO group was lower than those in SHAM group and 2VO+ LCAR group (both P<0.05). Electron microscopic observation of mitochondrial morphology showed that the Flameng score of mitochondrial damage in the hippocampus of rats in 2VO group (2.82±0.17) was higher than those in SHAM group (0.25±0.07) and 2VO+ LCAR group (1.76±0.09) (both P<0.05). (3) The density of synaptic vesicles in the hippocampus of rats in 2VO group ((289.09±22.41)/μm 2)was lower than those in SHAM group ((497.49±28.89)/μm 2)and 2VO+ LCAR group ((401.23±45.09)/μm 2) (both P<0.01). Western blot results showed that the relative levels of synaptic proteins NR2A/B, PSD95 and CREB in 2VO group were lower than those in SHAM group and 2VO+ LCAR group (all P<0.05). Immunofluorescence results showed that the relative level of CREB expression in hippocampal subregions and cortex in 2VO group was lower than those in SHAM group and 2VO+ LCAR group (both P<0.01). Conclusion:L-carnitine can improve spatial learning and memory dysfunction in rats with chronic cerebral hypoperfusion, which are related with promoting ATP production and protecting mitochondrial morphology, and promoting synaptic vesicle synthesis and synaptic protein expression.

Effects of high frequency flash stimulation on the capacity of learning memory and autonomous exploratory in young rats / 中华行为医学与脑科学杂志

Objective:To investigate the effects of 40 Hz and 70 Hz frequency flash stimulation on the ability of learning memory and autonomous exploratory in young rats.Methods:Twenty-seven SPF grade male SD rats aged 19-21 days were divided into control group (Ctr group), 40 Hz group and 70 Hz group with 9 in each group according to the random number table.The rats in Ctr group were not given flash stimulation, while rats in the 40 Hz and 70 Hz group were received 40 Hz, 70 Hz flash stimulation (1.5 h/d for 39 days), respectively.The Morris water maze experiment was used to assess the learning and memory ability of rats, and the open field experiment was used to evaluate the ability of autonomous exploratory of rats.Nissl staining was used to assess the morphology of Nissl bodies in the hippocampus CA1 region of the rats.The local field potentials (LFPs) collected from the primary visual cortex (V1 area) region by electrophysiological experiments was used to verify the synchronization of flash evoked neural oscillations.SPSS 23.0 software was used for statistical analysis.The repeated measures ANOVA and one-way ANOVA were used to analyze normal distribution measurement data, and LSD and Tamhane tests were used for further pairwise comparison.The Kruskal-Wallis test was used for non-normal distribution measurement data.Results:(1) The flash stimulation of 40 Hz and 70 Hz both can effectively caused synchronization of neural oscillations in the primary visual cortex of healthy young rats.(2) The results of repeated measures ANOVA analysis showed that there was no interaction effect of grouping and time in the escape latency of young rats in the Morris water maze positioning navigation phase( F=1.326, P>0.05 ). The escape latency had time main effect ( F=40.025, P<0.05), but no grouping main effect ( F=2.039, P>0.05). With the increase of learning days, the escape latency of young rats in each group decreased significantly.There was no interaction effect of grouping and time in the total distance of young rats ( F=2.029, P>0.079). It had time main effect ( F=32.052, P<0.05), but not grouping main effect ( F=2.390, P>0.05) on total distance.With the increase of learning days, the total distance of young rats in each group significantly shortened.On the 6th day of the Morris water maze experiment, there was no statistically significant difference among groups in terms of time in the target quadrant and the number of crossing platforms ( F=2.511, 0.802, both P>0.05). The results of the open field experiment showed that the total distance traveled in the center of young rats in each group was statistically significant ( H=8.935, P<0.05), the total distance traveled in the center in the 70 Hz group (3.80 (2.25, 6.93) m)was significantly longer than that in the 40 Hz group (0.80 (0.72, 1.46) m), P<0.05). The percentage of time spent in the center was statistically significant in the three groups ( H=11.050, P<0.05). Young rats in the 70 Hz group spent significantly higher percentage of time in the center(3.20(2.43, 8.30)) than those in the 40 Hz group (0.95 (0.37, 1.06 ), P<0.05 ). (3) Nissl staining results showed that Nissl bodies in the hippocampal CA1 area of young rats in Ctr, 40 Hz and 70 Hz group were all arranged neatly and tightly, no edema was found in the surrounding stroma, and no obvious inflammatory cell infiltration was found. Conclusion:70 Hz frequency flash stimulation may promote the ability of learning memory and autonomous exploratory of young rats.

Roles and mechanisms of metformin in improving cognitive dysfunction induced by chronic cerebral hypoperfusion in rats / 中华行为医学与脑科学杂志

Objective:To explore the roles and mechanisms of metformin in the improvement of cognitive dysfunction induced by chronic cerebral hypoperfusion in rats.Methods:Total 82 SD male rats (SPF grade) aged 3-4 months were randomly divided into four groups: sham operation control group (Con group, n=15), sham operation with metformin treatment group (Met group, n=20), 2-vessel occlusion group (2VO group, n=22), and 2-vessel occlusion with metformin administration group (2VO+ Met group, n=25). The chronic cerebral hypoperfusion model was established by bilateral common carotid artery ligation, and the carotid arteries of rats in Con group and Met group were only separated without ligation.After 2VO operation, rats in 2VO+ Met group and Met group were given metformin solution in drinking water at a dose of 100 mg/kg per day for 4 weeks.After 4-week continuous intervention with metformin, Morris water maze was performed to test the spatial cognitive function of the rats, in vivo electrophysiological technology was used to detect the long-term potential of the rats, and enzyme-linked immunosorbent assay(ELISA) was used to detect the concentrations of inflammatory factor tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and interleukin-6(IL-6) in the hippocampus.The density of dendritic spines of hippocampal neurons was observed by Golgi staining, and the synaptic structure of hippocampal neurons, especially the vesicle density, was observed by transmission electron microscopy.SPSS 16.0 software was used for statistical analysis.Repetitive measurement ANOVA was used for the escape latency data of 7 days repeated learning training in water maze.One-way ANOVA was used for the comparison of other data among multiple groups, and Dunnett's t test was used for further pairwise comparison. Results:Morris water maze results showed that during 7 days of learning training, the time and group interaction for escape latency was not significant in the 4 groups of rats ( F=0.93, P>0.05), but the time main effect ( F=25.90, P<0.05) and group main effect ( F=13.20, P<0.05) were significant.Morris water maze test showed that from the 3rd to 7th day, the escape latencies in 2VO group were significantly longer than those in Con group and 2VO+ Met group(all P<0.05). The short-term memory of rats was detected after 1 day of rest.The results showed that the escape latency in 2VO group was significantly longer than that in Con group and 2VO + Met group( P<0.01). The retention time and crossing times in the platform area of 2VO rats were less than those in Con group and 2VO + Met group ( P<0.01). Electrophysiological results showed that the relative field excitatory postsynaptic potential slope of 2VO group (1.29±0.09) was significantly lower than that in Con group (2.07±0.09) and 2VO + Met group (1.69±0.08)( P<0.01). ELISA results showed that TNF-α level in hippocampal tissue of 2VO group was significantly higher than that in Con group and 2VO+ Met group; IL-1β and IL-6 levels in hippocampal tissue of 2VO group were significantly higher than those in Con group and 2VO + Met group.Density of dendritic spines in hippocampal neurons of 2VO group was significantly lower than that in Con group and 2VO+ Met group.The density and proportion of immature dendritic spines in hippocampal neurons of 2VO group were significantly higher than those in Con group and 2VO + Met group.Synaptic vesicle density of neurons in CA1 area of hippocampus in 2VO group ((230.29±19.44) vescicles/μm 2) was significantly lower than that in the Con group ((414.52±13.17) vescicles/μm 2) and 2VO+ Met group ((313.19±12.42) vescicles/μm 2). Conclusion:Metformin can reduce neuroinflammation of hippocampus with chronic cerebral hypoperfusion and improve synaptic plasticity and cognitive dysfunction.It may have potential application value in the treatment of vascular cognitive dysfunction.