Therapeutic effect of transplantation of bone marrow mesenchymal stem cells co-cultured with bone marrow M2 macrophages on a rat model of liver cirrhosis / 临床肝胆病杂志

ObjectiveTo investigate the effect of transplantation of bone marrow mesenchymal stem cells （BMSCs） co-cultured with bone marrow-derived M2 macrophages （M2-BMDMs）， named as BMSCM2， on a rat model of liver cirrhosis induced by carbon tetrachloride （CCl4）/2-acetaminofluorene （2-AAF）. MethodsRat BMDMs were isolated and polarized into M2 phenotype， and rat BMSCs were isolated and co-cultured with M2-BMDMs at the third generation to obtain BMSCM2. The rats were given subcutaneous injection of CCl4 for 6 weeks to establish a model of liver cirrhosis， and then they were randomly divided into model group （M group）， BMSC group， and BMSCM2 group， with 6 rats in each group. A normal group （N group） with 6 rats was also established. Since week 7， the model rats were given 2-AAF by gavage in addition to the subcutaneous injection of CCl4. Samples were collected at the end of week 10 to observe liver function， liver histopathology， and hydroxyproline （Hyp） content in liver tissue， as well as changes in the markers for hepatic stellate cells， hepatic progenitor cells， cholangiocytes， and hepatocytes. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the N group， the M group had significant increases in the activities of serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） （P<0.01）； compared with the M group， the BMSC and BMSCM2 groups had significant reductions in ALT and AST （P<0.01）， and the BMSCM2 group had significantly better activities than the BMSC group （P<0.05）. Compared with the N group， the M group had significant increases in Hyp content and the mRNA and protein expression levels of alpha-smooth muscle actin （α-SMA） in the liver （P<0.01）； compared with the M group， the BMSC and BMSCM2 groups had significant reductions in Hyp content and the expression of α-SMA （P<0.05）， and the BMSCM2 group had a significantly lower level of α-SMA than the BMSC group （P<0.01）. Compared with the N group， the M group had significant increases in the mRNA expression levels of the hepatic progenitor cell markers EpCam and Sox9 and the cholangiocyte markers CK7 and CK19 （P<0.01） and significant reductions in the expression levels of the hepatocyte markers HNF-4α and Alb （P<0.01）； compared with the M group， the BMSC and BMSCM2 groups had significant reductions in the mRNA expression levels of EpCam， Sox9， CK7， and CK19 （P<0.05） and significant increases in the mRNA expression levels of HNF-4α and Alb （P<0.05）， and compared with the BMSC group， the BMSCM2 group had significant reductions in the mRNA expression levels of EpCam and CK19 （P<0.05） and significant increase in the expression level of HNF-4α （P<0.05）. ConclusionM2-BMDMs can enhance the therapeutic effect of BMSCs on CCl4/2-AAF-induced liver cirrhosis in rats， which provides new ideas for further improving the therapeutic effect of BMSCs on liver cirrhosis.

Efecto regenerador gástrico del consumo de Petroselinum sativum L. (perejil) en ratas con gastritis inducida por etanol

El objetivo del presente trabajo fue determinar el efecto regenerador gástrico del consumo de Petroselinum sativum L. (perejil) en ratas con gastritis inducida por etanol. Se realizó un estudio analítico, experimental clásico, transversal, prospectivo. Se trabajó con 36 ratas Wistar machos (250 ± 30 g.p.c.) distribuidas aleatoriamente en 6 grupos (n=6). Los grupos II-VI fueron sometidos a ayuno de 24 horas para inducirles úlcera gástrica administrándoles 10 mL/kg.p.c. de etanol al 70% vía orogástrica. Después de una hora, se procedió a sacrificar al grupo II para observar el daño ulceroso en el estómago. Después, se elaboró el extracto acuoso de hojas frescas de perejil (EAHP) y se administró a los demás grupos el siguiente tratamiento por vía orogástrica durante 3 días: grupo III, 10 mL/kg.p.c. de solución de NaCl al 0,9%; y EAHP a los grupos IV-VI (150, 300 y 600 mg/kg.p.c., respectivamente). Enseguida, las ratas fueron sometidas a ayuno de 24 horas para luego ser sacrificadas por desnucamiento. Posteriormente, se les realizó una laparotomía para la extracción del estómago. El EAHP generó mayor producción de moco gástrico en las dosis de 300 mg/kg.p.c. con 78,03% y de 600 mg/kg.p.c. con 80,52% (p<0,05). Esto concordó con lo observado histológicamente en la mucosa gástrica, mostrando solo signos de inflamación de la submucosa en los grupos que consumieron EAHP (IV-VI), en comparación con necrosis fibrinoide de los grupos que no lo consumieron (II y III). En conclusión, el consumo de EAHP tiene un efecto regenerador gástrico en ratas con gastritis inducida por etanol.

Our objective is to determine the gastric regenerative effect of Petroselinum sativum L. (parsley) consumption in rats with ethanolinduced gastritis. We developed an analytical, experimental, classical, cross-sectional, prospective study. We worked with 36 male Wistar rats (250 ± 30 g.p.c.) randomly distributed into 6 groups (n=6). Groups II-VI were subjected to a 24-hour fast to induce gastric ulcer by administering 10 mL/kg.p.c. of 70% ethanol via orogastric. After one hour, group II was sacrificed to observe the ulcerative damage in the stomach. Afterward, the aqueous extract of fresh parsley leaves (EAHP) was prepared, and the following treatment was administered to the other groups through the orogastric route for 3 days: group III, 10 mL/kg.p.c. 0.9% NaCl solution; and EAHP to groups IV-VI (150, 300, and 600 mg/Kg.p.c., respectively). The rats were then fasted for 24 hours before being sacrificed by breaking their necks. Subsequently, a laparotomy was performed to extract the stomach. The EAHP generated greater production of gastric mucus in the doses of 300 mg/kg.p.c. with 78.03% and 600 mg/kg.p.c. with 80.52% (p<0.05). This was consistent with what was observed histologically in the gastric mucosa, showing only signs of inflammation of the submucosa in the groups that consumed EAHP (IV-VI), compared with fibrinoid necrosis in the groups that did not consume it (II and III). In conclusion, the consumption of EAHP has a gastric regenerative effect in rats with ethanol-induced gastritis.

Action of enfuvirtide on the pregnancy of albino rats ( Rattus norvegicus albinus, Rodentia, Mammalia ): biological assay and functional and histological analyses of exposed maternal-fetal organs

ABSTRACT Objective To assess the effects of enfuvirtide on pregnancy in albino rats and their fetuses. Methods Forty pregnant EPM 1 Wistar rats were randomly allocated into four groups: control (E) (distilled water twice/day), G1 (4mg/kg/day enfuvirtide), G2 (12mg/kg/day enfuvirtide), and G3 (36mg/kg/day enfuvirtide) groups. On the 20th day of gestation, the rats were anesthetized and subjected to cesarean section. Their blood was collected for laboratory analysis, and they were sacrificed. The offspring's fragments of their kidneys, liver, and placentas and the maternal rats' fragments of their lungs, kidneys, and liver were separated in the immediate postpartum period for light microscopy analysis. Results No maternal deaths occurred. In the second week at the end of pregnancy, the mean weight of the G3 Group was significantly lower than that of the G2 Group (p=0.029 and p=0.028, respectively). Analyzing blood laboratory parameters, the G1 Group had the lowest mean amylase level, and the G2 Group had the lowest mean hemoglobin level and the highest mean platelet count. In the morphological analysis, there were no changes in organs, such as the kidneys and liver, in both the maternal rats and offspring. Three maternal rats in the G3 Group had pulmonary inflammation in the lungs. Conclusion Enfuvirtide has no significant adverse effects on pregnancy, conceptual products, or functional alterations in maternal rats.

Articular mobilization promotes improvement in functional and inflammatory parameters in a gouty arthritis model

ABSTRACT Objective Gouty arthritis is characterized by painful inflammation due to the deposition of monosodium urate crystals in joint tissues. Despite available treatments, many patients experience ineffective management and adverse effects. This study evaluated a manual therapy protocol involving passive joint mobilization at the peak of inflammation in a gouty arthritis model using functional and inflammatory parameters. Methods Twenty male Wistar rats, 12 weeks old, were divided into two groups (n=10 each): Gouty Arthritis and Control Groups, which were further subdivided into treated and untreated groups (n=5 each). The Gouty Arthritis Group received intraarticular knee injection of 50µL of monosodium urate crystals, while the Control Group received 50µL of phosphate buffered saline. The treatment involved a 9-minutes session of grade III joint mobilization (according to Maitland). Nociception, grip strength, and edema were evaluated before induction (EV0), 7 hours after assessment (EV1), immediately after treatment (EV2), and 1 hour after treatment (EV3). The animals were euthanized, and synovial fluid was collected to analyze leukocyte migration. Results The model mimicked the signs of the Gouty Arthritis Group, with a decrease in the threshold of nociception and strength and an increase in edema and leukocyte count. The mobilization protocol significantly increased the nociceptive threshold and grip strength and reduced edema; however, it did not reverse the increase in leukocyte count. Conclusion Our results suggest that mobilization promotes analgesia and may modulate the inflammatory process owing to reduced edema and subtle attenuation of cell migration, which contributes to strength gain.

Indicadores morfométricos de la corteza cerebral frontal de gazapos de ratas Wistar con diabetes pregestacional

Introducción: La diabetes es una enfermedad que afecta el embarazo provoca complicaciones fetales; dentro de ellas son frecuentes las malformaciones congénitas. Por la imposibilidad práctica y ética de estudiar este proceso en gestantes es imprescindible realizar estudios experimentales empleando procedimientos morfométricos para determinar si la diabetes afecta el neurodesarrollo. Objetivo: Caracterizar morfométricamente la sustancia gris de gazapos de ratas Wistar normales y con diabetes mellitus pregestacional. Métodos: Se realizó un estudio experimental básico de serie de casos a 20 gazapos de ratas Wistar de los cuales 10 eran descendientes de diabetes pregestacional. Se caracterizaron indicadores morfométricos del tejido nervioso como espesor de corteza e indicadores nucleares como el perímetro. Resultados: La media de la altura de la sustancia gris cortical mostró un valor de 1,224 ± 303,7 μm para el grupo control y 1,014 ± 376,0 para los casos, al aplicar el test de diferencias de medias se encontró diferencia significativa (p ≤ 0,05) a favor del grupo control. Los valores de la media del perímetro nuclear en el grupo control fue de 42,80 ± 7,23 μm y en el grupo experimental el promedio fue de 39,68 ± 6,52 μm, al aplicar el test de diferencias de medias se encontró diferencia significativa (p ≤ 0,05) a favor del grupo control al presentar mayor perímetro nuclear. Conclusiones: El mayor espesor cortical y perímetro nuclear correspondió al grupo control evidenciándose el efecto deletéreo de la diabetes mellitus en el neurodesarrollo.

Introduction: Diabetes is a disease that affects pregnancy causing fetal complications; within them congenital malformations are frequent. Due to the practical and ethical impossibility of studying this process in pregnant women, it is essential to carry out experimental studies using morphometric procedures to determine if diabetes affects neurodevelopment. Objective: To characterize morphometrically the gray matter of kits from normal Wistar rats and those with pregestational diabetes mellitus. Methods: A basic experimental study of a series of cases was carried out on 20 young Wistar rats, of which 10 were descendants of pregestational diabetes. Morphometric indicators of the nervous tissue were characterized as thickness of the cortex and nuclear indicators such as perimeter. Results: The average height of the cortical gray matter showed a value of 1.224 ± 303.7 μm for the control group and 1.014 ± 376.0 μm for the cases. When applying the mean difference test, a significant difference was found (p ≤ 0.05) in favor of the control group. The values of the measurement of the nuclear perimeter in the control group was 42.80 ± 7.23 μm and in the experimental group the average was 39.68 ± 6.52 μm. When applying the mean different test, a significant difference was found (p ≤ 0.05) at favor of control group presenting greater nuclear perimeter. Conclusions: The greatest cortical thickness and nuclear perimeter corresponded to the control group, evidencing the deleterious effect os diabetes mellitus on neurodevelopment.

Obesity influences the development of bisphosphonate-induced osteonecrosis in Wistar rats

Abstract Medication-related osteonecrosis of the jaw (MRONJ) is characterized by bone exposure for more than eight weeks in patients who have used or been treated with antiresorptive or antiangiogenic drugs, without a history of radiation therapy or metastatic diseases in the jaws. Obesity is associated with changes in periodontal tissues and oral microbiota that are linked to bone alterations. This study aimed to analyze the influence of obesity on the development of bisphosphonate-induced osteonecrosis. The experiment randomly and simply divided 24 male Wistar rats (Rattus norvegicus) into four groups: healthy, with osteonecrosis, obese, and obese with osteonecrosis (n=6 per group). Osteonecrosis was induced through weekly intraperitoneal injection for eight weeks at a dose of 250 µg/kg of zoledronic acid in a 4 mg/5 mL solution, combined with trauma (exodontia). Obesity was induced through a high glycaemic index diet. Each group was qualitatively and quantitatively evaluated regarding the development of models and pathological anatomy of the lesions. The results were expressed in mean percentage and standard deviation and statistically analyzed using one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test, with a significance level of 5% (p<0.05) to establish differences found between the groups. Animals in the osteonecrosis group and the obese with osteonecrosis group presented larger necrosis areas (averages: 172.83±18,19 µm2 and 290.33±15,77 µm2, respectively) (p<0,0001). Bone sequestration, hepatic steatosis, and increased adipocyte size were observed in the obese group (average: 97.75±1.91 µm2) and in the obese with osteonecrosis group (average: 98.41±1.56 µm2), indicating greater tissue damage in these groups (p<0,0001). All parameters analyzed (through histological, morphometric, and murinometric analyses) increased for the obese and obese with osteonecrosis groups, suggesting a possible influence of obesity on the results. However, further studies are needed to confirm the role of obesity in the possible exacerbation of osteonecrosis and understand the underlying mechanisms.

Evaluation of the effect of intrathecal GM1 in 24, 48, and 72 hours after acute spinal cord injury in rats

Abstract Objective The aim of this study was to evaluate the best timing and feasibility of intrathecal application of sodium monosialoganglioside (GM1) after spinal cord contusion in Wistar rats as an experimental model. Methods Forty Wistar rats were submitted to contusion spinal cord injury after laminectomy. The animals were randomized and divided into four groups: Group 1 - Intrathecal application of GM1 24 hours after contusion; Group 2 - Intrathecal application of GM1 48 hours after contusion; Group 3 - intrathecal application of GM1 72 hours after contusion; Group 4 - Sham, with laminectomy and intrathecal application of 0.5 mL of 0.9% saline solution, without contusion. The recovery of locomotor function was evaluated at seven different moments by the Basso, Beattie, and Bresnahan (BBB) test. They were also assessed by the horizontal ladder, with sensory-motor behavioral assessment criteria, pre-and postoperatively. Results This experimental study showed better functional scores in the group submitted to the application of GM1, with statistically significant results, showing a mean increase when evaluated on known motor tests like the horizontal ladder and BBB, at all times of evaluation (p < 0.05), especially in group 2 (48 hours after spinal cord injury). Also, fewer mistakes and slips over the horizontal ladder were observed, and many points were achieved at the BBB scale analysis. Conclusion The study demonstrated that the intrathecal application of GM1 after spinal cord contusion in Wistar rats is feasible. The application 48 hours after the injury presented the best functional results.

Establishment of a rat model of acute radiation esophagitis / 中华放射肿瘤学杂志

Objective:To establish Wistar rat models of acute radiation esophagitis, and observe the histopathological changes at different time points after modeling.Methods:Wistar rats were locally irradiated with different doses of 6 MV X-rays, and the rats were sacrificed on the 3 rd, 5 th, 7 th, and 14 th days after irradiation. The full-length esophagus tissue was taken for paraffin embedding, sectioning, and hematoxylin and eosin (HE) staining for pathological assessment. The pathological changes of the esophagus of the rats were observed at the 3 rd, 5 th, 7 th, and 14 th days after 25 Gy and 30 Gy irradiation. The changes of daily dietary intake of rats in different irradiation groups within 1-2 weeks after radiation exposure were observed. Results:No rat died in two groups after being irradiated with 25 Gy and 30 Gy rays. All the rats in the 30 Gy group had esophagus injury. On the 7 th day, the degree of injury was the most serious, with a pathological score of 5.00±0.75 and a food intake of 0 g. On the 14 th day, the degree of injury was relieved, and the food intake was restored to the level before irradiation. Conclusions:The Wistar rat model of acute radiation esophagitis can be established by a single dose of 6 MV X-ray 30 Gy irradiation to the esophagus. The 7 th day after irradiation is an ideal observation time for the acute injury phase, which is gradually alleviated after the 7 th day. The time can be chosen from 7-14 days after irradiation as the observation point for the healing repair phase.

Daño en la pared arterial de las ratas Wistar sometidas a dieta hiperglucídica / Arterial wall damage in Wistar rats subjected to a hyperglycemic diet

RESUMEN Introducción: Los modelos experimentales en animales proporcionan una valiosa información para comprender los procesos fisiopatológicos de las lesiones de los vasos sanguíneos y sus consecuencias. Objetivo: Analizar los cambios histológicos y morfométricos que se observaron en la aorta abdominal de las ratas Wistar sometidas a una dieta hiperglucídica. Métodos: Se formaron aleatoriamente dos grupos experimentales de 10 animales cada uno. El grupo control alimentado con dieta estándar para la especie, y el grupo experimental alimentado con dieta estándar más sacarosa al 35 %, como agua de bebida desde el destete hasta las 20 semanas de vida. El estudio se realizó en muestras de aorta fijadas y procesadas por la técnica clásica de inclusión en parafina y coloreadas con las técnicas de hematoxilina - eosina y Verhoeff. Se realizó la descripción de las capas de la pared arterial y la determinación de variables morfométricas en cada lámina histológica. Resultados: Las ratas Wistar pertenecientes al grupo experimental desarrollaron modificaciones incipientes en la pared arterial de la aorta abdominal, las cuales corresponden con la presencia de tumefacción en la célula endotelial y vacuolización en la célula muscular lisa vascular, así como marcada desorganización de las fibras elásticas y musculares de la capa media. Las variables morfométricas que mostraron diferencias significativas entre los grupos fueron el grosor de la túnica media y el cociente media/ lumen. Conclusiones: La pared media resultó ser la capa más afectada demostrándose el efecto nocivo de la dieta hiperglucídica en la pared arterial.

ABSTRACT Introduction: experimental animal models provide valuable information to understand the physiological and pathological processes of blood vessel injuries and their consequences. Objective: to analyze the histological and morphometric changes observed in the abdominal aorta of Wistar rats subjected to a hyperglycemic diet. Methods: two experimental groups of 10 animals each were randomly formed. The control group was fed with a standard diet for this species, and the experimental group was fed with a standard diet plus 35% sucrose, as drinking water from weaning to 20 weeks of life. The study was performed on aortic samples fixed and processed by the classic paraffin embedding technique and stained with the hematoxylin-eosin and Verhoeff techniques. The description of the layers of the arterial wall and the determination of morphometric variables in each histological slide were made. Results: the Wistar rats belonging to the experimental group developed incipient changes in the arterial wall of the abdominal aorta, which correspond to the presence of endothelial cell swelling and vacuolation in the vascular smooth muscle cell, as well as marked disorganization of the muscle and elastic fibers of the middle layer. The morphometric variables that showed significant differences between the groups were the thickness of the tunica media and the media/lumen ratio. Conclusions: the middle wall turned out to be the most affected layer, demonstrating the harmful effect of the hyperglycemic diet on the arterial wall.

Comparative Evaluation of MTA Sealers Biomineralization Ability: an in Vivo Study / Avaliação Comparativa da Capacidade de Biomineralização de Cimentos MTA: Estudo in Vivo

This research aimed to evaluate the biomineralization induced by different concentrations of MTA Flow® and compare it to MTA Angelus®. Fifteen male Wistar rats received subcutaneous implants containing the materials to be tested (MTA Flow® at putty, thick, and thin consistencies and MTAAngelus®) and empty tubes (control). After 7, 40 and 90 days, the animals were euthanized, and the implants were removed with the surrounding tissue. The presence of biomineralization was analyzed in light microscope by Von Kossa technique. The statistica l differences were considered for p < 0.05. Calcification areas were present in all the MTA Flow® and MTA Angelus® groups. In the control group, no mineralized areas were observed. MTA Angelus® and thin-MTA Flow® showed significant reduction in calcification as time went by. A significant increase in areas with calcification, proportional to the exposure time, was observed in putty-MTA Flow® and thick-MTA Flow®. MTA Angelus® and thin-MTA Flow® showed significantly higher calcification than thick-MTA Flow® in the shortest exposure time. Analysis of putty-MTA Flow® showed significantly higher calcification areas than MTA Angelus® and thin-MTA Flow® in the longest exposure time. MTA Flow® stimulated mineralization, which has varied according to the concentration. Besides, in longer periods, MTA Flow® biomineralization performance was higher than MTAAngelus®, especially in highest concentration. (AU)

Esse estudo teve como objetivo avaliar a biomineralização induzida por diferentes concentrações do MTA Flow® e compará-la ao MTA Angelus®. Quinze ratos Wistar machos receberam implantes subcutâneos contendo os materiais a serem testados (MTA Angelus® e MTA Flow® nas consistências pastosa, espessa e fluida) e tubos vazios (controle). Após 7, 40 e 90 dias, os animais foram eutanasiados e os implantes foram removidos juntamente com o tecido circundante. A presença de biomineralização foi analisada em microscópio de luz pela técnica de Von Kossa. Diferenças estatísticas foram consideradas para valores de p < 0,05. Áreas de calcificação estavam presentes em todos os grupos do MTA Flow® e MTA Angelus®. No grupo controle não foram observadas áreas mineralizadas. O MTA Angelus® e o MTA Flow® fluido apresentaram redução significativa na quantidade de calcificação ao longo do tempo. Um aumento significativo na quantidade de áreas calcificadas, proporcional ao tempo de exposição, foi observado no MTA Flow® pastoso e no MTA Flow® espesso. O MTA Angelus® e o MTA Flow® fluido apresentaram calcificação significativamente maior do que o MTA Flow® espesso no menor período de exposição. Análises contento o MTA Flow® pastoso demonstraram áreas de calcificação significativamente maior do que o MTA Angelus® e MTA Flow® fluido no maior tempo de exposição. O MTA Flow® induziu a formação de áreas mineralizadas, que variou de acordo com a concentração do cimento. Em períodos mais longos, o MTA Flow® apresentou desempenho superior ao MTA Angelus®, principalmente quando utilizado na maior concentração. (AU)

Relación del ácido fólico y los defectos del tubo neural en ratas Wistar / Relationship of folic acid and neural tube defects in Wistar rats

RESUMEN Introducción: los defectos del cierre del tubo neural son anomalías del sistema nervioso central superadas únicamente por los defectos cardíacos. En Pinar del Río existe un aumento de interrupciones de embarazo por diagnóstico de este defecto, se desconoce si los niveles de ácido fólico en las gestantes influyen en su incidencia. Objetivo: corroborar la relación entre los defectos del cierre del tubo neural en las crías de ratas Wistar y las dosis de ácido fólico administrada en las ratas Wistar gestadas. Métodos: se realizó un estudio experimental en ratas Wistar, se formaron cinco grupos de dos ratas hembras, dos que recibieron dosis de ácido fólico de 100 y 200 microgramos (µg) antes y durante la gestación, dos que recibieron 100 y 200 µg durante la gestación y un grupo control que no recibió ninguna dosis. La muestra estuvo constituida por 212 ratas recién nacidas, sacrificadas bajo normas internacionales. Se extrajo médula y cerebro para observar las anomalías, se tomaron fotomicrografía de los cortes realizados. El nivel de significación para todas las comparaciones estadísticas fue α ≤ 0,05. Resultados: se observaron en total trece espinas bífidas, once en el grupo control (26,19 %) y dos (4,65 %) en el de 200 µg durante la gestación, al comparar el grupo control respecto a los grupos tratados, la variable masa corporal presentaron significación estadística. Se observó que la presencia de espina bífida fue significativa en la hembra con respecto al macho. Conclusiones: se demostró la necesidad esencial de la ingestión de ácido fólico antes y durante la gestación en ratas Wistar y la asociación de espina bífida y sexo, el sexo femenino fue el predominante.

ABSTRACT Introduction: neural tube closure defects are anomalies of the central nervous system surpassed only by cardiac defects. In Pinar del Rio province there is an increase of pregnancy interruptions due to diagnosis of this defect, and it is not known if folic acid levels in pregnant women influence its incidence. Objective: to corroborate the relationship between neural tube closure defects in Wistar rats' pups and the doses of folic acid administered in pregnant Wistar rats. Methods: an experimental study was carried out in Wistar rats, forming five groups of two female rats; two of them received doses of folic acid of 100 and 200 micrograms (µg) before and during gestation, two that received 100 and 200 µg during gestation and a control group that did not receive any dose. The sample consisted of 212 newborn rats, sacrificed under international standards. Marrow and brain were extracted to observe the anomalies, and photomicrographs were taken of the sections made. The significance level for all statistical comparisons was α ≤ 0,05. Results: a total of thirteen spina bifida were observed, eleven in the control group (26,19 %) and two (4,65 %) in the 200 µg group during gestation, when comparing the control group with the treated groups, the body mass variable presented statistical significance. It was observed that the presence of bifid spina was significant in the female with respect to the male. Conclusions: the essential need of folic acid ingestion before and during gestation in Wistar rats and the association of spina bifida and sex were confirmed, being predominant in the female sex.

Alteraciones genitales en la infección por leptospiras en ratas Wistar gestadas / Genital alterations in leptospiras infection in pregnant Wistar rats

RESUMEN Introducción: la leptospirosis en Cuba se presenta de forma endémica, y a veces de forma epidémica, por lo que es un problema de salud. Su efecto sobre los genitales internos, en gestantes, ha sido poco tratado por los investigadores. Objetivo: describir las principales alteraciones en los genitales internos de ratas Wistar gestadas infestadas por leptospiras. Métodos: se realizó un estudio experimental en ratas Wistar preñadas: siete grupos con tres animales cada uno: un control negativo y seis casos con dosis de una concentración bacteriana de 300 millones de Leptospiras por mL; se inocularon dos mililitros a cada animal al cuarto día de gestadas por 17 meses. Se tomaron los genitales internos de todas las ratas el día 20 de la gestación, estos fueron procesados en parafina y teñidos con hematoxilina/eosina. Resultados: se observaron alteraciones en útero, ovarios y trompas; el mayor número de lesiones se presentaron en útero mientras que en ovarios y trompas se visualizaron la misma cantidad de lesiones. Conclusiones: la leptospirosis infectó los genitales internos con la producción de quistes, necrosis, abscesos, ooforitis y microhemorragias en ovarios; congestión vascular y hemorragias en las trompas uterinas y endometritis, hemorragias, miometritis, piometristis, perimetritis y congestión vascular en el útero. Predominaron las alteraciones congestivas y hemorrágicas.

ABSTRACT Introduction: leptospirosis in Cuba occurs endemically, and sometimes epidemically, making it a health problem. Its effect on internal genitalia in pregnant women has been little treated by researchers. Objective: to describe the main alterations in the internal genitalia of pregnant Wistar rats infested by leptospiras. Methods: an experimental study was carried out in pregnant Wistar rats: 7 groups with three animals each: 1 negative control and 6 cases with doses of a bacterial concentration of 300 million Leptospira per mL; each animal was inoculated with two milliliters on the fourth day of pregnancy for 17 months. The internal genitalia of all rats were taken on day 20 of gestation, processed in kerosene and stained with hematoxylin/eosin. Results: alterations were observed in the uterus, ovaries and tubes; the greatest number of lesions was present in the uterus while the same number of lesions was visualized in the ovaries and tubes. Conclusions: leptospirosis infected the internal genitalia producing cysts, necrosis, abscesses, oophoritis and microhemorrhages in the ovaries; vascular congestion and hemorrhages in the uterine tubes and endometritis, hemorrhages, myometritis, pyometristis, perimetritis and vascular congestion in the uterus. Congestive and hemorrhagic alterations predominated.

Prostatic alterations associated to early weaning and its relation with cocoa powder supplementation. Experimental study in adult wistar rats

ABSTRACT Early weaning can predispose the offspring to greater risk of developing chronic diseases in adulthood. It is believed that the consumption of functional foods is able to prevent these effects. The aim of this study is to evaluate the effects of maternal and postnatal cocoa powder supplementation on body mass, metabolism, and morphology of the prostate of early weaned Wistar rats. The animals were divided into four experimental groups according to lactation time (21 or 18 days, n=6, each) as follows: control group (C), cocoa control group (CCa), early weaning group (EW), and cocoa early weaning group (EWCa). The animals were euthanized at 90 days of age. Serum biochemical analysis and prostate histomorphometric evaluation were performed. The animals supplemented with cocoa powder were heavier than their respective controls (p <0.05), although with no difference in food intake among the groups. Likewise, these same groups showed a reduction in the serum glucose in relation to C and EW groups (p <0.0001). With respect to the prostate, there was no difference in smooth muscle and lumen area densities, while the EW group had a lower epithelial height and a higher percentage of mast cells than the C group (p <0.05). On the other hand, the EWCa group managed to reverse these parameters, leveling with the controls. Early weaning resulted in hyperglycemia and important morphological changes in the prostate. In contrast, dietary supplementation with cocoa powder attenuated these effects on the metabolism and prostatic histoarchitecture, proving to be a good nutritional treatment strategy.

Regenerating effect of hyaluronic acid on circumferential bone defects in wistar albino rats / Efecto regenerador del ácido hialurónico en defectos óseos circunferenciales en ratas albinas wistar

Objective: To determine the regenerating effect of hyaluronic acid on circumferential bone defects in albino Wistar rats. Material and Methods: An experimental type study was designed and carried out with 15 albino male Wistar rats, 4 months old and weighing between 250 and 350 grams. Two circumferential bone defects 3mm in diameter and 0.8mm deep were created in the calvaria of the parietal bone (on both sides of the midline). One defect was filled with a demineralized bone matrix (control group); while the other defect was filled with the combination of a demineralized bone matrix plus hyaluronic acid (experimental group). Five experimental rats were euthanized at 30, 60 and 90 days after surgery and they were histologically evaluated following the parameters proposed by Heiple. Results: The experimental group presented a better degree of bone regeneration at 30 and 60 postoperative days. Conclusion: Hyaluronic acid is effective in bone regeneration of circumferential bone defects.

Objetivo: Determinar el efecto regenerador del ácido hialurónico en defectos óseos circunferenciales en ratas albinas Wista. Material y Métodos: Se diseñó un estudio de tipo experimental y se trabajó con 15 ratas albinas Wistar (todas macho) de 4 meses de edad y con un peso entre 250 a 350 gr. Se crearon en todas 2 defectos óseos circunferenciales de 3mm de diámetro y 0.8 mm de profundidad en la calota del hueso parietal (a ambos lados de la línea media). Un defecto fue rellenado con una matriz ósea desmineralizada (grupo control); mientras que el otro defecto fue rellenado con la combinación de una matriz ósea desmineralizada más el ácido hialurónico (grupo experimental). Se realizó la eutanasia a 05 ratas de experimentación a los 30, 60 y 90 días postquirúrgicos y se evaluaron histológicamente siguiendo los parámetros propuestos por Heiple. Resultados: El grupo experimental presentó un mejor grado de regeneración ósea en los 30 y 60 días postoperatorios. Conclusiones: El ácido hialurónico es eficaz en la regeneración ósea de defectos óseos circunferenciales.

Morphofunctional study on the effects of passive smoking in kidneys of rats / Estudo morfofuncional sobre os efeitos do tabagismo passivo em rins de ratos

ABSTRACT Objective To analyze whether passive inhalation of cigarette smoke causes morphological, structural, and functional changes in kidneys of rats. Methods Wistar rats, aged eight weeks, weighing on average 260g, were divided into Control Group and Smoking Group. Each group was subdivided into four groups of ten animals for morphofunctional analysis, in a period of seven and 28 days. The Smoking Group was exposed to smoke of 40 cigarettes per day, at certain times and in automated equipment for cigarette burning, called smoking machine (SM-MC-01). After the exposure period, urine and blood samples were collected for the functional analyses, and the kidneys were dissected and submitted to histological procedures for morphoquantitative analyses. Results After exposure of animals of the Smoking Group, the following were observed: lower weight gain; lower water and feed intake; decreased renal weight, diameter, and volume; reduction in cortical thickness and glomerular volume density; decrease in glomerular and capsular diameter; increase in mesangial density; decreased urine volume; increased levels of glucose, serum creatinine and microalbuminuria; decreased urinary creatinine levels and creatinine clearance rate. Conclusion Passive smoking negatively influences renal morphology and glomerular filtration rate, with effects similar to those described in the literature regarding active smoking.

RESUMO Objetivo Analisar se a inalação passiva da fumaça do cigarro proporciona alterações morfológicas, estruturais e funcionais nos rins de ratos. Métodos Ratos Wistar, com oito semanas de idade, pesando, em média, 260g, foram divididos em Grupo Controle e Grupo Tabagista. Cada grupo foi subdividido em quatro grupos de dez animais para análise morfofuncional, em um período de sete e 28 dias. O Grupo Tabagista foi exposto à fumaça de 40 cigarros por dia, em horários determinados e equipamento automatizado de queima de cigarros, denominado smoking machine (SM-MC-01). Após o período de exposição, foram coletadas amostras de urina e sangue para as análises funcionais, e os rins foram dissecados e submetidos a procedimentos histológicos para análises morfoquantitativas. Resultados Após a exposição dos animais do Grupo Tabagista, observou-se menor ganho de peso; menor consumo de água e ração; menor peso, diâmetro e volume renal; redução em espessura cortical e densidade de volume glomerular; diminuição no diâmetro glomerular e capsular; aumento na densidade mesangial; volume urinário diminuído; níveis aumentados de glicose, creatinina sérica e microalbuminúria; níveis reduzidos de creatinina urinária e redução da taxa de depuração da creatinina. Conclusão O tabagismo passivo influencia negativamente na morfologia renal e na taxa de filtração glomerular, com efeitos semelhantes aos descritos na literatura em relação ao tabagismo ativo.

Effects of photobiomodulation on wound contraction in rats undergoing doxorubicin extravasation: a histomorphometric analysis / Efectos de la fotobiomodulación en la contracción de heridas en ratones sometidos a la extravasación de doxorrubicina: análisis histomorfométrico / Efeitos da fotobiomodulação na contração de feridas em ratos submetidos ao extravasamento de doxorrubicina: análise histomorfométrica

ABSTRACT Objective: To analyze wound contraction and histomorphometric pattern of lesions in Wistar rats undergoing doxorubicin extravasation. Method: Sixty adult female rats were used, divided into four groups of fifteen animals: Group 1 (Control, without antidote); Group 2 (Hyaluronidase); Group 3 (Photobiomodulation), and Group 4 (Hyaluronidase + Photobiomodulation). Doxorubicin 1mg (0.5 ml) was applied subcutaneously on the animals' back, inducing the wound. Macroscopic and morphometric evaluation of the lesions was performed every two days for 28 days. On the 30th day, euthanasia was performed and the material was collected for histological evaluation. Results: The animals in the photobiomodulation and photobiomodulation + Hyaluronidase groups presented regeneration tissue with neovascularization and acute inflammation, with improvement in wound healing, which did not occur in the other groups. The contraction rates were better in those treated with photobiomodulation and photobiomodulation + Hyaluronidase, with healing percentages of 76.6% and 72.1%, respectively. Conclusion: The combination of photobiomodulation (660 nm-1 J) with topical hyaluronidase (65 UTR) proved to be effective in the process of wound healing due to extravasation of doxorubicin, and can be incorporated into the practice of clinical oncology.

RESUMEN Objetivo: Analizar la contracción de heridas y patrón histomorfométrico de las lesiones en ratones Wistar sometidos a la extravasación de doxorrubicina. Método: Fueron utilizadas 60 ratas adultas, divididas en cuatro grupos de quince animales: Grupo 1 (Control sin antídoto); Grupo 2 (Hialuronidasa); Grupo 3 (Fotobiomodulación) y Grupo 4 (Hialuronidasa + Fotobiomodulación). Se aplicó 1 mg (0,5 ml) de doxorrubicina en el subcutáneo del dorso de los animales, induciendo la herida. Realizada evaluación macroscópica y morfométrica de las lesiones cada dos días por 28 días. En el 30° día se realizó la eutanasia y recolección del material para evaluación histológica. Resultados: Los animales de los grupos fotobiomodulación y fotobiomodulación+ Hialuronidasa presentaron tejido de regeneración con neovascularización e inflamación aguda, con mejoría de la cicatrización de las heridas, lo que no ocurrió en otros grupos. Las tasas de contracción fueron mejores en los tratados con fotobiomodulación y fotobiomodulación + Hialuronidasa, con porcentaje de cicatrización de 76,6% y 72,1%, respectivamente. Conclusión: la combinación de la fotobiomodulación (660nm-1J) con hialuronidasa tópica (65 UTR) demostró ser más eficaz en el proceso de cicatrización de heridas por extravasación de doxorrubicina, lo que posibilitó adoptarla en la práctica de la oncología clínica.

RESUMO Objetivo: Analisar a contração de feridas e padrão histomorfométrico das lesões em ratos Wistar submetidos ao extravasamento de doxorrubicina. Método: Utilizados 60 ratos fêmeas adultos, divididos em quatro grupos de quinze animais: Grupo 1 (Controle, sem antídoto); Grupo 2 (Hialuronidase); Grupo 3 (Fotobiomodulação) e Grupo 4 (Hialuronidase + Fotobiomodulação). Aplicado 1 mg (0,5 ml) de doxorrubicina no subcutâneo do dorso dos animais, induzindo a ferida. Realizada avaliação macroscópica e morfométrica das lesões a cada dois dias por 28 dias. No 30º dia foi realizada a eutanásia e coleta do material para avaliação histológica. Resultados: Os animais dos grupos fotobiomodulação e fotobiomodulação+Hialuronidase apresentaram tecido de regeneração com neovascularização e inflamação aguda, com melhora na cicatrização das feridas, o que não ocorreu nos outros grupos. As taxas de contração foram melhores nos tratados com fotobiomodulação e fotobiomodulação + Hialuronidase, com percentual de cicatrização de 76,6% e 72,1%, respectivamente. Conclusão: A combinação da fotobiomodulação (660 nm-1 J) com hialuronidase tópica (65 UTR) mostrou-se eficaz no processo de cicatrização de feridas por extravasamento de doxorrubicina, podendo ser incorporada à pratica da oncologia clínica.

Effects of photobiomodulation therapy on functional recovery, angiogenesis and redox status in denervated muscle of rats / Efeitos da terapia de fotobiomodulação na recuperação funcional, na angiogênese e no estado redox em músculo desnervado de ratos

ABSTRACT Objective: To evaluate the effects of photobiomodulation therapy in redox status, angiogenesis marker - vascular endothelial growth factor - and in the functional recovery in denervated muscle. Methods: A total of 32 female Wistar rats underwent a crush injury and were randomly divided into four groups: Light Emitting Diode Group 2 and Control Group 2 (muscle collected 2 days after injury), and Light Emitting Diode Group 21 and Control Group 21 (muscle collected 21 days afterinjury). Light Emitting Diode Group 2 and Light Emitting Diode Group 21 received two and ten light emitting diode applications (630±20nm, 9J/cm2, 300mW), respectively, and the Control Group 2 and Control Group 21 did not receive any treatment. The function was evaluated by grasping test at four moments (pre-injury, 2, 10 and 21 post-injury days). The flexor digitorum muscle was collected for analysis of immunolocalization of vascular endothelial growth factor and redox parameters. Results: Functional improvement was observed at the second and tenth post-injury day in treated groups compared to control (p<0.005). The muscle tissue of treated groups presented higher immunohistochemical expression of vascular endothelial growth factor. Photobiomodulation therapy decreased the oxidative damage to lipid in Light Emitting Diode Group 2 compared to Control Group 2 (p=0.023) in the denervated muscle. Conclusion: Photobiomodulation therapy accelerated the functional recovery, increased angiogenesis and reduced lipid peroxidation in the denervated muscle at 2 days after injury.

RESUMO Objetivo: Avaliar os efeitos da terapia de fotobiomodulação no estado redox, no marcador de angiogênese - fator de crescimento endotelial vascular - e na recuperação funcional do músculo desnervado. Métodos: Um total de 32 ratas Wistar foi submetido a uma lesão por esmagamento e dividido aleatoriamente em quatro grupos: Grupo Diodo Emissor de Luz 2 e Grupo Controle 2 (músculo coletado 2 dias após a lesão), além do Grupo Diodo Emissor de Luz 21 e do Grupo Controle 21 (músculo coletado 21 dias após a lesão). Grupo Diodo Emissor de Luz 2 e Grupo Diodo Emissor de Luz 21 receberam duas e dez aplicações de diodo emissor de luz (630±20nm, 9J/cm2e 300mW), respectivamente, e Grupo Controle 2 e Grupo Controle 21 não receberam tratamento. A função foi avaliada pelo teste de preensão em quatro momentos (pré-lesão, 2, 10 e 21 dias após a lesão). O músculo flexor dos dedos foi coletado para análise dos parâmetros redox e da imunolocalização do fator de crescimento endotelial vascular. Resultados: Houve melhora funcional no segundo e décimo dia pós-lesão nos grupos tratados em comparação aos controles (p<0,005). O tecido muscular dos grupos tratados apresentou maior expressão imuno-histoquímica do fator de crescimento endotelial vascular. A terapia de fotobiomodulação diminuiu o dano oxidativo aos lipídeos no Grupo Diodo Emissor de Luz 2 comparado ao Grupo Controle 2 (p=0,023) no músculo desnervado. Conclusão: A terapia de fotobiomodulação acelerou a recuperação funcional, aumentou a angiogênese e reduziu a peroxidação lipídica no músculo desnervado 2 dias após a lesão.

Ibuprofen increases the serum Omentin levels in rats after abdominal surgery

SUMMARY AIMS Omentin is an adipokine primarily produced by visceral adipose tissue and its reduced levels have been shown to be associate with worse metabolic outcomes. We aimed to study the effects of preoperative ibuprofen on postoperative omentin levels in rats after surgery. METHODS Forty-eight albino Wistar rats, 6 in each of 8 groups according to the surgical procedure (laparotomy, laparotomy plus ibuprofen (IBU), nephrectomy, nephrectomy plus IBU, hepatectomy, hepatectomy plus IBU, splenectomy and splenectomy plus IBU). The Omentin levels of the groups were postoperatively analyzed. RESULTS The mean omentin was significantly higher in the laparotomy plus IBU group compared to the laparotomy group (p<0.001). Mean Omentin was significantly higher in the hepatectomy plus IBU group compared to the hepatectomy group (p=0.01). Mean Omentin was significantly higher in the nephrectomy plus IBU group compared to the nephrectomy group (p=0.001). CONCLUSION We suggest that preoperative ibuprofen may enhance circulating levels of Omentin, which has beneficial effects in trauma and inflammation settings in subjects that undergo minor or major abdominal surgery.

RESUMO OBJETIVOS A omentina é uma adipocina produzida principalmente pelo tecido adiposo visceral e níveis reduzidos dela foram associados a piores desfechos metabólicos. Nosso objetivo foi estudar os efeitos do uso pré-operatório do ibuprofeno nos níveis pós-operatórios da omentina em ratos. METODOLOGIA Quarenta e oito ratos Wistar albinos foram divididos em 8 grupos (6 em cada), de acordo com o procedimento cirúrgico: laparotomia, laparotomia e ibuprofeno (IBU), nefrectomia, nefrectomia e IBU, hepatectomia, hepatectomia e IBU, esplenectomia, e esplenectomia e IBU. Os níveis de omentina dos grupos foram analisados após a cirurgia. RESULTADOS A omentina média foi significativamente maior no grupo de laparotomia e IBU do que no grupo de laparotomia (p<0,001). A omentina média foi significativamente maior no grupo de hepatectomia e IBU do que no grupo de hepatectomia (p = 0,01). A omentina média foi significativamente maior no grupo de nefrectomia e IBU do que no grupo de nefrectomia (p = 0,001). CONCLUSÃO Sugerimos que o uso pré-operatório de ibuprofeno pode aumentar os níveis circulantes de omentina, que têm efeitos benéficos em um contexto de trauma e inflamação em indivíduos submetidos cirurgia abdominal.

The anti-liver fibrosis effect of Kangxian Ruangan prescription: A cell study based on hepatic sinusoidal capillarization / 临床肝胆病杂志

ObjectiveTo investigate the anti-liver fibrosis effect and mechanism of action of Kangxian Ruangan prescription based on hepatic sinusoidal capillarization. MethodsA total of 16 male Wistar rats were divided into Kangxian Ruangan prescription group and control group, with 8 rats in each group. The rats in the Kangxian Ruangan prescription group were given crude drug 40 g/kg body weight by gavage, and those in the control group were given normal saline by gavage. Serum containing Kangxian Ruangan prescription was prepared, and then primary liver sinusoidal endothelial cells were treated with the serum containing Kangxian Ruangan prescription or the control serum. Western blot was used to measure the protein expression of the vascular endothelial markers cluster of differentiation 31 (CD31) and von Willebrand factor (vWF), and immunocytochemistry was used to measure the expression of CD31 and vWF in cells. Scanning electron microscopy was used to observe the change in the fenestrae of liver sinusoidal endothelial cells. The t-test was used for comparison of continuous data between two groups. ResultsKangxian Ruangan prescription had no marked influence on the morphology of liver sinusoidal endothelial cells. The liver sinusoidal endothelial cells in the control group had a small number of narrow fenestrae with an occluded structure, while those in the Kangxian Ruangan prescription group had an increased number of large fenestrae with a clear structure. The results of immunocytochemistry showed that compared with the control group, the Kangxian Ruangan prescription group had significant reductions in the expression of CD31 (0.069±0015 vs 0.115±0.011, t=4.257, P＜0.05) and vWF (0.092±0.009 vs 0.132±0.014, t=4.274, P＜0.05). The results of Western blot showed that compared with the control group, the Kangxian Ruangan prescription group had significant reductions in the protein expression of CD31 (0.334±0.029 vs 0.448±0.013, t=6.245, P＜0.01) and vWF (0.560±0.047 vs 0709±0.045, t=3.966, P＜0.05). ConclusionKangxian Ruangan prescription can significantly resist hepatic sinusoid capillarization and thus prevent the progression of liver fibrosis.