Maternal separation and early social enrichment increase anxietyrelated behaviours of adolescent rats in the elevated plus maze / La separación materna y el enriquecimiento social temprano aumentan los comportamientos relacionados con ansiedad de ratas adolescentes en el laberinto en cruz elevado

Abstract Introduction: Adverse or favourable rearing conditions early in life affect emotional response during adolescence. To study the effect of early rearing on emotional response, animal models such as maternal separation (MS) and social enrichment (SE) by community nesting have been useful. However, the comparison of the effect of MS and SE on anxiety-related behaviours in adolescent rats is unknown. Objective: This study aimed to assess the effects of MS and SE on the emotional response of adolescent rats exposed to the elevated plus-maze test (EPM). Method: Pregnant Wistar rats were randomly distributed into three groups according to the rearing condition. In the MS group, pups were separated daily from their dams for 180 minutes, from postnatal day (P) 2 to 14. In the SE group, two females that gave birth synchronously were housed in a cage with a litter of eight pups. Females from control standard housing (SH) were individually housed and kept with their offspring until weaning (P23). On P32, anxiety-related measures were evaluated using an EPM. Results: MS and SE increase anxiety-related behaviours and locomotion in rats exposed to the EPM. SE had sex-dependent effects on anxiety-related measures, increasing vertical activity in females and horizontal activity in males. MS but not SE increased body weight gain in female rats. Conclusion: Adverse or favourable rearing conditions early in life may result in an increased anxiety phenotype in the EPM during adolescence. It is likely that the favourable effect of SE depends on the number of dams per communal nest.

Resumen Introducción: Las condiciones de crianza adversas o favorables en etapas tempranas de la vida afectan la respuesta emocional durante la adolescencia. Para estudiar el efecto de la crianza temprana sobre la respuesta emocional, han sido útiles modelos animales como la separación materna (SM) y el enriquecimiento social (ES) mediante anidamiento comunitario. Sin embargo, se desconoce la comparación del efecto de la SM y el ES sobre las conductas relacionadas con la ansiedad en ratas adolescentes. Objetivo: Este estudio pretende evaluar los efectos de la SM y el ES sobre la respuesta emocional de ratas adolescentes expuestas al test del laberinto en cruz elevado (EPM). Método: Las ratas Wistar preñadas se distribuyeron aleatoriamente en tres grupos según la condición de crianza. En el grupo SM, las crías fueron separadas diariamente de sus madres durante 180 minutos, desde el día postnatal (P) 2 hasta el 14. En el grupo ES, dos hembras que parieron de forma sincronizada se alojaron en una caja con una camada de ocho crías. Las hembras del alojamiento estándar de control (SH) se alojaron individualmente y se mantuvieron con sus crías hasta el destete (P23). En P32, se evaluaron las medidas relacionadas con la ansiedad mediante un EPM. Resultados: La SM y el ES aumentan las conductas relacionadas con la ansiedad y la locomoción en las ratas expuestas al EPM. El ES tuvo efectos dependientes del sexo en las medidas relacionadas con ansiedad, aumentando la actividad vertical en las hembras y la actividad horizontal en los machos. La SM, pero no el ES, incrementó la ganancia de peso corporal en las ratas hembra. Conclusiones: Condiciones de crianza adversas o favorables en etapas tempranas de la vida pueden dar lugar a un fenotipo de ansiedad aumentado en el EPM durante la adolescencia. Es probable que el efecto favorable del ES dependa del número de hembras por nido comunitario.

Evaluation of nephrotoxicity effects of the methanol leaf extract of A. angustifolia in Wistar rats

Background: Agave angustifolia is a common traditional remedy in localities for disease treatment. This study focussed on evaluating the nephrotoxicity activity of the methanol extract of A. angustifolia leaf. Methods: Twenty-five Wistar rats of both sex were randomly shared into five groups. Group 1 received 10 mL/kg distilled water, group 2 gentamicin 80mg/kg, extract groups (3, 4 and 5) were placed on 100 mg/kg, 200 mg/kg and 400 mg/kg of A. angustifolia leaf extract administered orally through an orogastric tube for 14 days. The rats were sacrificed using chloroform, and their kidneys were harvested, weighed and immediately fixed in 10% buffered formalin for histological analysis and blood samples were collected by cardiac puncture for biochemical and haematological analysis. Results: Results showed there was no marked difference in the levels of the packed cell volume (PCV) in all the concentrations of the extract as well as the controls. The gentamicin group showed a remarkable rise in the serum urea and creatinine level when compared to both the control and extract groups. Similar effects were observed in the 100 mg/kg and 200 mg/kg of the extract. However, severe kidney injury was observed in the group treated with 400 mg/kg of the extract. Conclusions: Despite the beneficial potential of A. angustifolia, it also exhibited toxic effects on the kidney thereby causing significant damage to the kidney morphology at higher doses. The damages inflicted in these tissues are dose-dependent. Therefore A. angustifolia should be taken in low doses within shortest period of time.

Compressão intermitente imediata sobre anastomose vascular: Uma nova técnica para prevenir trombose? / Immediate intermittent compression over vascular anastomosis: a new technique to prevent thrombosis

Introdução: Este estudo tem o objetivo de avaliar o efeito da compressão intermitente imediata sobre anastomoses arteriais microcirúrgicas em comparação com compressão fixa e com utilização isolada de irrigação com soro fisiológico e heparina em laboratório experimental. Método: 12 ratos Wistar foram aleatoriamente divididos em três grupos para terem suas artérias femorais seccionas e anastomosadas de forma término-terminal, para comparação de patência com 30 minutos e 7 dias. Grupo I: foi realizada compressão intermitente imediata sobre a anastomose por 60 segundos; grupo II: uma compressão fixa foi mantida imediatamente após a anastomose, também por 60 segundos; grupo III, após o término da anastomose, não foi feita nenhuma intervenção adicional. Além da avaliação da patência, os animais foram pesados e medidos os diâmetros arteriais operados. Resultados: 24 artérias femorais foram abordadas. As médias de peso inicial dos ratos dos grupos I, II e III foram, respectivamente, de 243,8g, 254,6g e 260,4g, enquanto as finais foram de 264,4g, 281g e 282,1g (p<0,001). O diâmetro médio das artérias abordadas foi, respectivamente, de 0,89mm, 0,88mm e 0,90mm, e os tempos de anastomoses em minutos, de 25,6, 24,5 e 24,5, respectivamente; As patências finais após 7 dias foram, respectivamente, de 62,5% (p=0,07), 25% (p=0,48) e 50% (p=0,13). Conclusão: A compressão intermitente imediata pode ser realizada ao término de anastomoses arteriais microcirúrgicos sem prejuízo na patência final do procedimento.

Introduction: This study aims to evaluate the effect of immediate intermittent compression on microsurgical arterial anastomoses in comparison with fixed compression and only observation in an experimental laboratory. Methods: The two femoral arteries of twelve male Wistar rats were sectioned and reanastomosed to compare patency at 30 minutes and 7 days. Group I: immediate intermittent compression was performed over the anastomosis for 60 s; group II: a fixed compression was maintained immediately after the anastomosis for 60 s; group III: after completion of the anastomosis, no additional intervention was performed. In addition to the patency assessment, the animals were weighed and the operated arterial diameters were measured. Results: Twenty-four femoral arteries were examined. Initial average weights of the rats in groups I, II, and III were 243.8g, 254.6g, and 260.4g, respectively, while the final weights were 264.4g, 281g, and 282.1g (p<0.001), respectively; mean diameter of the approached arteries was 0.89, 0.88, and 0.90mm, respectively, and the anastomoses (time in minutes) were 25.6, 24.5, and 24.5, respectively; final patencies after 7 days were 62.5% (p=0.07), 25% (p=0.48), and 50% (p=0.13), respectively. Conclusion: Immediate intermittent compression can be performed at the end of microsurgical arterial anastomoses without affecting the final patency of the procedure.

Effect of Swimming at Low Temperature on Uncoupling Protein Gene Expression in Skeletal Muscle of Wistar Rats / Efecto de la Natación a Baja Temperatura sobre la Expresión del Gen de la Proteína de Desacoplamiento en el Músculo Esquelético de Ratas Wistar

SUMMARY: As the economy develops and living standards improve, overweight and obesity are increasingly prevalent. Currently, weight-loss medications are primarily administered orally or intravenously, which can result in poor targeting, low bioavailability, frequent administration, and high toxicity and side effects. The study aimed to address these challenges by preparing polylactic acid- polyethylene glycol staple fibers that carry the browning drug pioglitazone hydrochloride using electrostatic spinning and freeze-cutting techniques. Animal experiments were conducted to test the effectiveness of these fibers. Additionally, the study investigated the expression of uncoupling protein genes in rats exposed to different water temperatures by measuring changes in serum urea nitrogen and mRNA expression levels of skeletal muscle uncoupling protein genes. The physiological and genetic effects of low-temperature swimming exercise on changes in energy metabolism in rats were also analyzed at both the individual and molecular levels. The results revealed that serum urea nitrogen remained more stable in hypothermic swimming rats compared to rats in the swimming group. Furthermore, the study observed an induced up-regulation of uncoupling proteins in the skeletal muscle of Wistar rats in response to external temperature stimulation, and the expression of mRNA for skeletal muscle uncoupling proteins significantly increased as the temperature decreased. And the prepared short nanofibers also had a significant promotive effect on uncoupling protein gene, COX7A1, while suppressing the expression of lipogenic gene.

A medida que la economía se desarrolla y los niveles de vida mejoran, el sobrepeso y la obesidad son cada vez más frecuentes. Actualmente, los medicamentos para bajar de peso se administran principalmente por vía oral o intravenosa, lo que puede resultar en una mala focalización, baja biodisponibilidad, administración frecuente y alta toxicidad y efectos secundarios. El estudio tuvo como objetivo abordar estos desafíos mediante la preparación de fibras cortadas de ácido poliláctico y polietilenglicol que transportan el fármaco pardo clorhidrato de pioglitazona mediante técnicas de hilado electrostático y liofilización. Se realizaron experimentos con animales para probar la eficacia de estas fibras. Además, el estudio investigó la expresión de genes de proteínas desacopladoras en ratas expuestas a diferentes temperaturas del agua midiendo los cambios en el nitrógeno ureico sérico y los niveles de expresión de ARNm de genes de proteínas desacopladoras del músculo esquelético. También se analizaron los efectos fisiológicos y genéticos del ejercicio de natación a baja temperatura sobre los cambios en el metabolismo energético en ratas, tanto a nivel individual como molecular. Los resultados revelaron que el nitrógeno ureico sérico permaneció más estable en ratas nadadoras hipotérmicas en comparación con las ratas del grupo de natación. Además, el estudio observó una regulación positiva inducida de las proteínas desacopladoras en el músculo esquelético de ratas Wistar en respuesta a la estimulación de la temperatura externa, y la expresión de ARNm para las proteínas desacopladoras del músculo esquelético aumentó significativamente a medida que disminuía la temperatura. Además, las nanofibras cortas preparadas también tuvieron un efecto promotor significativo sobre el gen de la proteína de desacoplamiento, COX7A1, al tiempo que suprimieron la expresión del gen lipogénico.

Assessing the Impact of Annona muricata Aqueous Extract on Liver Enzymes in Wister Rats Exposed to Paracetamol Over Dose

Aims: This study examines the effect of the aqueous leaf extract of Annona Muricata on liver enzymes in wister rats using the spectrophotometric method. Methodology: 20 Wister rats were divided into groups A, B, C, and D, of 5 rats each, housed, fed, and the experimental groups administered treatments orally in distilled water (10 ml/kg) by means of orogastric cannula for 14 days. 500g of the plant powder was submerged for 72 hours 1500ml of distilled water. The sample was taken with a rotating evaporator, an electrical evaporator extraction apparatus. With water heated to 45癈, the solvent was removed. Before use, a paste-like sample was made and kept in a refrigerator at 4 degrees Celsius. The animals in group A were used as the control. The animals in group B were given feeds + 20mg of paracetamol, and the animals in group C were administered 100mg of paracetamol + silymarin. In comparison, the ones in group D were given 150mg of paracetamol + Annona muricata extracts. The animals were then sacrificed 48 hours following the final dose. A blood sample was collected, allowed to coagulate, and then the serum was separated, while the liver was excised and utilized for biochemical analyses.Results: The alanine aminotransferase activities (ul) recorded 9.0�71, 14.67�5, 13.50�58, 15.75�50 for animals in groups A, B, C, and D respectively with corresponding activities of aspartate aminotransferase (ul) of 7.25�50, 27.33�04, 20.00�71, 17.25�71. Results revealed a significant difference (p<0.05) in the activities of ALT and AST of all the treated groups. Thus, the aqueous leaf extract of Annona muricata could be hepato-protective. The silymarin could also be hepatoprotective, but it is less effective than Annona muricata. The untreated group, B, showed elevated levels of alanine aminotransferase and aspartate aminotransferase, confirming paracetamol's hepatotoxic activity. Conclusion: The ethanol leaf extract of Annona muricata decreased the activities of liver enzymes in male wister rats, suggesting its hepato-protective tendency. It also does not encourage body weight loss.

Effect of Murdannia edulis Faden roots on serum prolactin level and mammary glands of lactating female albino rats

Background: Murdannia edulis (Stokes) Faden (Myit-Cho in Myanmar) is a herb which has widely been used to induce or increase lactation by ethnic groups living in Tamu township of Sagaing division in Myanmar. This study aimed to explore the effect of Murdannia edulis on serum prolactin level and mammary glands of the lactating female rats. Methods: The type of the study was laboratory based experimental study. Thirty-six lactating dams were divided into six groups of 6 lactating rats in each group (n=6), namely group I (normal saline) as control, group II (5 mg/kg metoclopramide) as standard, groups III and IV (1 gm/kg and 2 gm/kg watery extract of Murdannia edulis), groups V and VI (1 gm/kg and 2 gm/kg ethanolic extract of Murdannia edulis), respectively. The extract and the drugs were orally administered from day 4 to 15 of lactation. At 16 days of lactation, the heart blood of lactating dams was collected under chloroform anaesthesia and serum prolactin level of dams was measured by using ELISA kit. The histological changes of mammary glands of the rats were also assessed. Results: The significant higher serum prolactin level were seen in ethanolic extract 2 gm/kg body weight treated group when compared to control. Histology of mammary glands in ethanolic extract treated rats showed stimulation of lobuloalveolar cell development. Conclusions: The lactogenic activity of the roots of Murdannia edulis may be due to promoting prolactin secretion and lobuloalveolar cell development of mammary glands.

Comparative Effects of Garcinia kola and Garlic Extracts on Some Liver and Haematological Functions in High Salt Diet-Induced Hypertensive Wistar Rats

Garcinia kola is commonly consumed in Nigeria in social ceremonies, while Garlic is commonly used as a spice in the preparation of certain foods. In this study, the protective effects of extracts of the plants against a high salt diet (8% NaCl) induced hypertension were investigated in rats. Extracts prepared from the plants were subjected to phytochemical screening for acute and sub-acute toxicity studies in rats. In the protective study against salt-induced hypertension thirty-five adult male rats assigned to seven groups of five rats were treated such that, group one served as the normal control group, and group two was the hypertensive control group. Groups three to six were administered graded doses of extracts respectively after induction. While group seven received a combination of the two extracts after induction. Treatment commenced after confirmation of hypertension and lasted for two weeks before animals were sacrificed to collect blood for biochemical and haematological analysis. Results obtained showed that G.kola and Garlic feeding produced in rats following liver and haematological assays as values of these parameters in the test groups did not significantly differ from control values (p<0.05).

Comparison of rapid dengue NS1, qualitative ELISA, and quantitative ELISA result

Background: Non-structural glycoprotein-1 (NS1) is a useful biomarker for early diagnosis of dengue fever. NS-1 antigen ELISA can be used for the early diagnosis of dengue fever in the acute stage. Quantitative methods are better for epidemic settings due to high false negative rates in qualitative ELISA. Methods: The study was initiated after approval from the institutional ethics council (IEC/DISS/17118). Study examined 280 patients with dengue symptoms who presented to the hospital's OPDs and IPDs. Patients were tested using qualitative ELISA, and those with Leptospira antibody, malaria, or Chikungunya IgM antibody were excluded. Age, gender, symptoms, comorbidities, total leucocyte count, platelet count, and risk category were all patient-related parameters. Patient-related parameters were recorded, and data was collected using Microsoft excel and analysed statistically. Results: Most patients aged 2-40 with male predominance had fever, chills, and body aches, 243 (86.8%) tested positive for ELISA NS1. Quantitative ELISA test showed a statistically significant correlation with rapid antigen NS1 result (p=0.015). Its AUC was 0.883 (p=0.0001), and its cut-off was (>109.1) with 96.9% sensitivity and 13.64% specificity. The AUC of quantitative ELISA NS1 against qualitative ELISA NS1 was 0.853 which was statistically significant (p<0.0001). At the cut-off >74.34, the test's sensitivity was 92.59% and specificity was 75.68%. Conclusions: Qualitative ELISA NS1 test is better than rapid antigen test for screening due to its higher specificity and similar sensitivity.

Phytochemical Composition and Toxicological Profiling of Curcuma longa (Turmeric) Root Extract in Rats

Background: Curcuma longa Linn (turmeric) is a common medicinal plant used in Africa for the treatment of diseases. In this study, its extract was evaluated to ascertain its phytochemical composition and toxicity in wistar rats. Methods: The prepared rhizome ethanol extract was first subjected to preliminary phytochemical evaluation and then lethal dose (LD50) test. In the sub-acute toxicity test, 20 adult wistar rats assigned to 4 groups of 5 rats each were administered oral graded doses of the extract. Group 1 served as control while groups 2, 3 and 4 received 250, 500 and 1000 mg/kg body weight of the extract over a period of 28 days, after which blood samples were collected for haematological and serum biochemical analyses. Results: Results obtained from the phytochemical analysis revealed the presence of saponins, tannins, phenolics, flavonoids, steroids, terpenoids, cardiac glycosides and alkaloids in extract, with alkaloids being most abundant (41.20+0.53 mg/100g) and terpenoids, the least (3.98+0.03 mg/100g). Lethal dose value for the extract was found to be >5000mg/kg body weight. Results of sub-acute toxicity evaluation of the extract showed no deleterious effect on liver and kidney function parameters with these parameters being not significantly different from their control values (p<0.05). Treatment with the extract also caused significant hypolipidemia (p<0.05) but increased the values of high-density lipoprotein cholesterol. The activities of antioxidant enzymes and red blood cell counts were also significantly increased following treatment with the extract, especially at higher dose levels. Conclusion: Therefore, we conclude that C. longa extract may be safe for use in the treatment of diseases when administered via the oral route.

Hematological, histopathological and oxidative stress responses to n-hexane extract of Terminalia catappa nuts in leukemia-induced Wistar rats

Background: Terminalia catappa nut possesses antioxidant and anticancer properties, but its effects on leukemia are unclear. This study investigates the effects of n-hexane extract of Terminalia catappa nut (TCN) on some hematological parameters, oxidative stress markers, and bone/spleen histopathology in a Wistar rat model of benzene-induced leukemia. Methods: Leukemia was induced in Wistar rats with 0.2 ml/kg Benzene solution and treated with 200, 400 or 800 mg/kg/day of Terminalia catappa nut extract (TCN) for 42 days and with 5-fluorouracil (20 mg/kg) via intraperitoneal injection twice a week for 6 weeks. Hematological parameters, antioxidant markers, and bone and spleen histology were analysed. Results: All TCN doses significantly lowered elevated WBCs by 32-53% and normalized RBC parameters compared to leukemic controls, mitigating cancer-induced anemia. TCN also exhibited potent antioxidant effects by enhancing SOD, GSH, and catalase while reducing MDA versus untreated rats. Bone marrow analysis revealed TCN conferred dose-dependent benefits on cellularity and architecture, reducing myeloid blasts and leukocyte infiltration. A near-normal bone microarchitecture was attained with the highest TCN dose. Similarly, TCN elicited marked improvements in splenic cytoarchitecture and attenuation of hypercellularity, lymphocytic infiltration and megakaryocytes compared to leukemic controls in a dose-dependent manner. Conclusions: Terminalia catappa nut extract demonstrated anti-leukemic, haematopoietic, antioxidant, and organ protective effects in leukemic Wistar rats induced with benzene solution, supporting its potential as an adjuvant therapeutic agent.

La desaferentación dental como factor etiológico de disfunciones gustativas en ratas Wistar machos / Dental deafferentation as an etiologic factor of taste dysfunction in male Wistar rats / A surdeferentação dentária como fator etiológico das disfunções gustativas em ratos Wistar machos

RESUMEN Objetivo : Evaluar la influencia de la desaferentación dental (DD) en el sentido del gusto de ratas Wistar machos mediante el test de reactividad gustativa (TRG). Materiales y métodos : Estudio experimental, siguiendo las directrices de ARRIVE 2.0, realizado en diez ratas Wistar. Se las aleatorizó y asignó en un grupo control o en un grupo experimental, y se implantaron cánulas para el TRG en ambos grupos. En el grupo experimental se realizó exodoncias de los tres molares superiores del lado derecho. Al tercer día se inició el TRG (día 1) mediante la infusión de 1 M de una sustancia dulce (ingestiva) y 3 mM de una sustancia amarga (aversiva), a razón de 1 mL en 1 minuto. Este TRG se repitió los días 7, 14 y 21. Se puntuó las respuestas ingestivas y aversivas durante 1 minuto. Los datos fueron procesados en el paquete estadístico SPSS v. 26. Se usó la prueba U de Mann-Withney para identificar las diferencias; y la magnitud de la diferencia se calculó mediante la r de Rosenthal. Resultados : Se obtuvo respuestas ingestivas a sacarosa el día 1 (p > 0,05); en los otros días se obtuvo respuestas diferentes: día 7 (p = 0,05), día 14 (p = 0,009) y día 21 (p = 0,009). Asimismo, se obtuvo respuestas aversivas a benzoato de denatonio (BD) los días 1, 7 y 21 (p > 0,05); esto fue diferente el día 14 (p = 0,05). Conclusiones : Se encontró una diferencia en las medianas de las respuestas ingestivas a sacarosa y aversivas a BD en ratas Wistar machos a consecuencia de la DD.

ABSTRACT Objective : To evaluate the influence of dental deafferentation (DD) on the sense of taste in male Wistar rats using the taste reactivity test (TRG). Materials and methods : An experimental study was conducted on ten Wistar rats, following ARRIVE 2.0 guidelines. They were randomized and assigned to a control or experimental group, and both groups' cannulae for the TRG were implanted. In the experimental group, exodontia of the three upper molars on the right side was performed. On the third day, GRT was started (day 1) by infusing 1 M of a sweet substance (ingestive) and 3 mM of a bitter substance (aversive) at a rate of 1 mL in 1 minute. This TRG was repeated on days 7, 14, and 21. Ingestive and aversive responses were scored for 1 minute. The data were processed in the SPSS v. 26 statistical package. The Mann-Whitney U test was used to identify differences, and the magnitude of the difference was calculated using Rosenthal's r. Results : Ingestive responses to sucrose were obtained on day 1 (p > 0.05); different responses were obtained on the other days: day 7 (p = 0.05), day 14 (p = 0.009), and day 21 (p = 0.009). Likewise, aversive responses to denatonium benzoate (BD) were obtained on days 1, 7, and 21 (p > 0.05); this was different on day 14 (p = 0.05). Conclusions : We found a difference in median ingestive responses to sucrose and aversive responses to BD in male Wistar rats due to DD.

RESUMO Objetivo : Avaliar a influência da surdeferentação dentária (DD) no sentido do paladar de ratos Wistar machos através do teste de reatividade gustativa (TRG). Materiais e métodos : Estudo experimental, seguindo as diretrizes ARRIVE 2.0, realizado em dez ratos Wistar. Estes foram aleatorizados e atribuídos a um grupo de controlo ou a um grupo experimental, tendo sido implantadas cânulas para TRG em ambos os grupos. No grupo experimental, foi efetuada a exodontia dos três molares superiores do lado direito. No terceiro dia, iniciou-se a TRG (dia 1) com a infusão de 1 M de uma substância doce (ingestiva) e 3 mM de uma substância amarga (aversiva), na velocidade de 1 mL em 1 minuto. Esse TRG foi repetido nos dias 7, 14 e 21. As respostas ingestivas e aversivas foram avaliadas durante 1 minuto. Os dados foram processados no pacote estatístico SPSS v. 26. O teste U de Mann-Withney foi usado para identificar diferenças; e a magnitude da diferença foi calculada usando o r de Rosenthal. Resultados : As respostas ingestivas à sacarose foram obtidas no dia 1 (p > 0,05); foram obtidas respostas diferentes no dia 7 (p = 0,05), no dia 14 (p = 0,009) e no dia 21 (p = 0,009). Também se obtiveram respostas aversivas ao benzoato de denatónio (BD) nos dias 1, 7 e 21 (p > 0,05); estas foram diferentes no dia 14 (p = 0,05). Conclusões : Foi encontrada uma diferença nas respostas ingestivas medianas à sacarose e nas respostas aversivas ao BD em ratos Wistar machos como resultado da DD.

Earthworm Protein Protects Vascular Endothelial Function in Spontaneously Hypertensive Rats via PI3K/Akt/Nrf2 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of earthworm protein on the expression of phosphatidylinositol 3-kinase/protein kinase B/nuclear factor E2-related factor 2 （PI3K/Akt/Nrf2） pathway in the aorta of spontaneously hypertensive rats （SHR） and explore mechanism of earthworm protein in treating hypertensive vascular endothelial dysfunction （VED）. MethodTen 10-week-old Wistar Kyoto （WKY） rats and fifty SHR rats were selected for a week of adaptive feeding. WKY rats were selected as the normal group， and fifty SHR rats were randomized according to body weight into model， valsartan （8×10-3 g·kg-1·d-1）， and high-， medium-， and low-dose （0.2， 0.1， 0.05 g·kg-1·d-1， respectively） earthworm protein groups. The normal and model groups were administrated with equal volume of double distilled water by gavage. During the drug intervention period， the general situations of rats in each group were observed and their blood pressure was monitored at specific time points every other week before and after administration. After 8 weeks of drug intervention， enzyme-linked immunosorbent assay was employed to measure the levels of angiotensin-Ⅱ （Ang-Ⅱ） and endothelin-1 （ET-1） in the serum of rats in each group. The corresponding kits were used to determine the levels of nitric oxide （NO）， malondialdehyde （MDA）， glutathione peroxidase （GPX）， superoxide dismutase （SOD）， and ferrous ion （Fe2+）. Hematoxylin-eosin （HE） staining was employed to observe the changes in the intima of the aorta. Fluorescence quantitative polymerase chain reaction （Real-time PCR） was employed to measure the mRNA levels of PI3K， Akt， Nrf2， heme oxygenase-1 （HO-1）， and glutathione peroxidase 4 （GPX4） in the aortic tissue. Western blotting was used to determine the protein levels of p-PI3K （Tyr467/199）， PI3K， p-Akt （Ser473）， Akt， Nrf2， HO-1， and GPX4 in the thoracic aorta. ResultCompared with the normal group， the model group had decreased body mass， increased irritability， severe endothelial damage， elevated blood pressure and serum levels of Ang-Ⅱ， ET1， MDA， and Fe2+ （P<0.01）， lowered NO level （P<0.01）， and down-regulated mRNA and protein levels of p-PI3K （Tyr467/199）， PI3K， p-Akt （Ser473）， Akt， Nrf2， HO-1， and GPX4 in the aortic tissue （P<0.01）. Compared with the model group， drug intervention caused no significant change in the body mass， calmed the rats， alleviated the endothelial damage， lowered blood pressure and serum levels of Ang-Ⅱ， ET1， MDA， and Fe2+ （P<0.01）， elevated the NO level （P<0.05）， and up-regulated the mRNA and protein levels of p-PI3K （Tyr467/199）， PI3K， p-Akt （Ser473）， Akt， Nrf2， HO-1， and GPX4 （P<0.05）. ConclusionThe earthworm protein can exert antihypertensive effects by ameliorating VED in SHR. Specifically， it may regulate the PI3K/Akt/Nrf2 signaling pathway to inhibit oxidative stress and ferroptosis.

Mechanism of Youguiwan Regulating BMP-2/Smad Signaling Pathway to Promote Bone Formation in Postmenopausal Rats with Osteoporosis / 中国实验方剂学杂志

ObjectiveTo observe the effects of Youguiwan on bone metabolism and bone morphogenetic protein-2 （BMP-2）/Smad signaling pathway in ovaries-removed rats with osteoporosis and study the mechanism of Youguiwan in the prevention and treatment of osteoporosis. MethodA postmenopausal rat model of osteoporosis was prepared by bilateral ovariectomy. The 40 female SD rats were randomly divided into five groups， including sham operation group， model group， alendronate sodium group （0.1 mg·kg-1）， and high-dose and low-dose （5.36 and 2.68 g·kg-1） groups of Youguiwan. The drug was given seven days after modeling， once a day for 12 weeks. After the treatment， the changes in femur tissue structure were observed by micro-CT， including bone mineral density （BMD）， bone volume/total volume （BV/TV）， trabecular number （Tb.N）， trabecular thickness （Tb.Th）， bone surface/bone volume （BS/BV）， and trabecular separation （Tb.Sp）. Ossification was observed by saffrane-solid green staining， and serum levels of bone metabolism markers， including bone alkaline phosphatase （BALP）， osteocalcin （BGP）， type Ⅰ procollagen amino terminal propeptide （PINP）， and tartrate-resistant acid phosphatase 5b （TRACP-5b）， were determined by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA expression levels of Runx2， BMP-2， and Smad1 in rat femur were detected by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the sham operation group， bone trabecula in the model group was sparse. BMD， BV/TV， Tb.N， and Tb.Th were decreased （P<0.05， P<0.01）. BS/BV （P<0.05） and Tb.Sp were increased. The content of BGP， BALP， PINP， and TRACP-5b in serum was significantly increased （P<0.01）. The mRNA and protein expressions of Runx2， BMP-2， and Smad1 in rat femur were significantly decreased （P<0.05， P<0.01）. Compared with the model group， the number of bone trabeculae in the high-dose and low-dose groups of Youguiwan was increased， and the bone microstructure was improved. BMD， BV/TV， Tb.N， and Tb.Th were increased significantly （P<0.05， P<0.01）， and BS/BV and Tb.Sp were increased. The content of bone metabolic markers decreased （P<0.05， P<0.01）. ConclusionYouguiwan has certain preventive and therapeutic effects on postmenopausal osteoporosis， and its mechanism may be related to promoting bone formation by regulating the BMP-2/Smad signaling pathway.

Changes in the total volume and contour density of oval cells in hepatic lobules of rats with carbon tetrachloride-induced hepatic fibrosis： A stereological study / 临床肝胆病杂志

ObjectiveTo quantitatively investigate the changes in the total volume and contour density of hepatic oval cells （HOC） in hepatic lobules of rats with carbon tetrachloride （CCl4）-induced hepatic fibrosis. MethodsA total of 11 healthy male Sprague-Dawley rats were randomly divided into control group with 5 rats and hepatic fibrosis group with 6 rats， and CCl4 and olive oil suspension were injected subcutaneously twice a week， 3 mL/kg each time. After five weeks of hepatic fibrosis modeling， five liver tissue blocks with a size of about 1 mm3 were randomly selected from the liver of each rat to prepare one Epon812 epoxy resin-embedded ultrathin section， and the stereological method and transmission electron microscopy were used for the quantitative analysis of the total volume and contour density of HOC in the hepatic lobules of rats. In addition， four liver tissue blocks with a thickness of 2 mm were randomly selected from the remaining liver of each rat to prepare two paraffin-embedded Masson staining sections， and the degree of liver fibrosis in each rat was qualitatively evaluated according to the Metavir staging criteria for liver fibrosis. The independent-samples t test was used for comparison of continuous data between groups. ResultsThe quantitative stereological analysis showed that the total volume of HOC in hepatic lobules was 15.40±7.63 mm3 in the control group and 146.80±114.00 mm3 in the liver fibrosis group， and compared with the control group， the total volume of HOC in hepatic lobules of rats in the liver fibrosis group was significantly increased by 8.53 times （t=-2.551， P=0.031）； the contour density of HOC in hepatic lobules was 56.20±40.40 in the control group and 566.50±317.00 in the liver fibrosis group， and compared with the control group， the contour density of HOC in hepatic lobules of rats in the liver fibrosis group was significantly increased by 9.08 times （t=-3.539， P=0.006）. Qualitative observation showed that liver fibrosis stage of rats reached stage Ⅱ-Ⅲ according to the Metavir scoring criteria， and massive proliferation of HOC was observed around the proliferation site of hepatic stellate cells in the perisinusoidal space of rats. ConclusionCCl4 induces significant proliferation of HOC in hepatic lobules of rats with liver fibrosis.

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.

The study on the effect of neuregulin-1 on myocardial injury in septic rats / 重庆医学

Objective To study the effect of neuromodulatory protein-1(NRG-1)in inhibiting sepsis induced myocardial injury and its mechanism.Methods The rat sepsis model was established by cecal ligation and puncture(CLP).SD rats were divided into the sham operation group,sepsis group,sepsis+NRG group(rhNRG,10 μg/kg).After 12,24 h of successful modeling,the heart and peripheral serum of the surviving rats in each group were taken respectively.The HE staining was used to observe the changes of cardiac tissue morphology and structure,and ELISA was used to detect the expression levels of creatine kinase(CK),crea-tine kinase MB isoenzyme(CK-MB),sensitive troponin Ⅰ(cTnⅠ)in serum,tumor necrosis factor-α(TNF-α)in cardiac tissue and IL-6 expression level;Western blot was used to detect the phosphorylation protein ki-nase B(p-Akt),phosphorylation glycogen synthase kinaseβ(p-GSK3β),B-cell lymphoma/leukemia-2(Bcl-2)and Bax protein expression in rat myocardial tissue.Results After 12,24 h of modeling,compared with the sham group,the expression levels of CK,CK-MB and cTnⅠ in serum,TNF-α,IL-6 and Bax protein in myocar-dial tissue in the sepsis group all were significantly increased(P＜0.05),while the expression levels of p-Akt and p-GSK3β in myocardial tissue were significantly decreased(P＜0.05).After 12,24 h of modeling,com-pared with the sepsis group,the expression levels of CK,CK-MB,cTn Ⅰ in the serum and the expression levels of TNF-α,IL-6 in the myocardial tissue of the sepsis+NRG group were significantly decreased;after 24 h of modeling,compared with the sepsis group,the expression level of Bax protein in myocardial tissue of the sep-sis+NRG group was decreased,while the p-Akt,p-GSK3β expression levels were increased(P＜0.05).The pathological results showed that compared with the sham operation group,the sepsis group produced signifi-cant lesions;compared with the sepsis group,the lesions in the sepsis+NRG group were alleviated.Conclusion The expression levels of related biomarkers in septic myocardial injury have change.NRG-1 could improve the cardiac function through Akt/GSK3β pathway,inhibit the related proinflammatory factors and reduce the myocardial tissue damage.

Toxicokinetics of MDMA and Its Metabolite MDA in Rats / 法医学杂志

Objective To investigate the toxicokinetic differences of 3,4-methylenedioxy-N-methylamphetamine(MDMA)and its metabolite 4,5-methylene dioxy amphetamine(MDA)in rats af-ter single and continuous administration of MDMA,providing reference data for the forensic identifica-tion of MDMA.Methods A total of 24 rats in the single administration group were randomly divided into 5,10 and 20 mg/kg experimental groups and the control group,with 6 rats in each group.The ex-perimental group was given intraperitoneal injection of MDMA,and the control group was given intraperi-toneal injection of the same volume of normal saline as the experimental group.The amount of 0.5 mL blood was collected from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.In the continuous administration group,24 rats were randomly divided into the experi-mental group(18 rats)and the control group(6 rats).The experimental group was given MDMA 7 d by continuous intraperitoneal injection in increments of 5,7,9,11,13,15,17 mg/kg per day,respectively,while the control group was given the same volume of normal saline as the experimental group by in-traperitoneal injection.On the eighth day,the experimental rats were randomly divided into 5,10 and 20 mg/kg dose groups,with 6 rats in each group.MDMA was injected intraperitoneally,and the con-trol group was injected intraperitoneally with the same volume of normal saline as the experimental group.On the eighth day,0.5 mL of blood was taken from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.Liquid chromatography-triple quadrupole tandem mass spectrometry was used to detect MDMA and MDA levels,and statistical software was employed for data analysis.Results In the single-administration group,peak concentrations of MDMA and MDA were reached at 5 min and 1 h after administration,respectively,with the largest detection time limit of 12 h.In the continuous administration group,peak concentrations were reached at 30 min and 1.5 h af-ter administration,respectively,with the largest detection time limit of 10 h.Nonlinear fitting equations for the concentration ratio of MDMA and MDA in plasma and administration time in the single-administration group and continuous administration group were as follows:T=10.362C-1.183,R2=0.974 6;T=7.397 3C-0.694,R2=0.961 5(T:injection time;C:concentration ratio of MDMA to MDA in plasma).Conclusions The toxicokinetic data of MDMA and its metabolite MDA in rats,obtained through single and continuous administration,including peak concentration,peak time,detection time limit,and the relationship between concentration ratio and administration time,provide a theoretical and data foundation for relevant forensic identification.

Effects of Shenyan 1 Prescription on renal fibrosis improvement in rats with unilateral ureteral obstruction based on TGF-β1/Smad3 signaling pathway / 国际中医中药杂志

Objective:To investigate the protective effects and mechanism of Shenyan 1 Prescription on renal fibrosis of unilateral ureteral obstruction (UUO) rats through TGF- β 1/Smad homologous 3 (Smad3) pathway regulating ferroptosis.Methods:Totally 48 male SD rats were divided into four groups: sham-operation group, UUO model group, and Shenyan 1 Prescription low-(10 drug/kg) , and high-dosage (20 crude drug/kg) groups according to random number table method, with 12 rats in each group. The UUO model was induced by the method of unilateral ureteral obstruction except for those sham-operation group. After modeling, rats received corresponding drugs or normal saline by gavage for 4 weeks, once per day. After 4 weeks, the body mass and the left kidney weight were measured. The 24 h urine protein and the levels of serum albumin (ALB), alanine aminotransferase (ALT), serum creatinine (SCr) and blood urea nitrogen (BUN) were detected by biochemical analysis method; the ROS level in renal tissue was measured using a chemical fluorescence assay kit, and the SOD and MDA levels in left renal tissue of rats were measured using ELISA method; the morphology of renal tissue and the specific blue staining of hemosiderin were observed using HE and Prussian blue staining methods, respectively; the expressions of transforming growth factor-β1 (TGF-β1), Smad3, glutathione peroxidase 4 (GPX4), and solute carrier family 1 member 5 (SLC1A5) were detected by Western blot.Results:Compared with the model group, the 24 h urinary protein excretion in Shenyan 1 Prescription high-dosage group decreased ( P<0.05), the serum ALB level increased ( P<0.05), the ALT level decreased ( P<0.05), and the expression of SLC1A5 in renal tissue decreased ( P<0.05); the left kidney weight/body decreased in Shenyan 1 Prescription low- and high-dosage groups ( P<0.05); the levels of serum ROS and MDA decreased ( P<0.05), and the activity of SOD significantly increased ( P<0.05); the expressions of TGF-β1 and Smad3 in renal tissue decreased ( P<0.05), and the expression of GPX4 increased ( P<0.05), and the renal pathological injury and ion deposition were improved. Conclusion:Shenyan 1 Prescription has a protective effect on the structure and function of renal tissues in UUO rats through regulating ferroptosis via inhibition of the TGF-β1/ Smad3 pathway to inhibit renal fibrosis of UUO rats.

Effects of Zhu Lian stimulant Type I acupuncture on the expression of brain-derived neurotrophic factor and its receptor TrkB and tissue homogenate cyclic adenosine phosphate in rats with diabetic bladder / 国际中医中药杂志

Objective:To observe the effects of Zhulian stimulant type Ⅰ acupuncture on the expression of brain-derived neurotrophic factor (BDNF) and its receptor TrkB and tissue homogenate cyclic adenosine phosphate (cAMP) in rats with diabetic bladder (DCP); To explore the mechanism of Zhulian stimulant type Ⅰacupuncture on DCP.Methods:Totally 50 SD rats were divided into control group, model group, Western medicine group, ordinary acupuncture group, Zhulian stimulant type Ⅰ acupuncture treatment group (acupuncture treatment group) according to random number table method, with 10 rats in each group. DCP rat model was prepared by intraperitoneal injection of streptozotocin (STZ), except for the control group. The Western medicine group was given mecobalamine for gavage; acupoints of "Zhongji", "Sanyinjiao", "Liechou" and "Taichong" were selected. The ordinary acupuncture group was treated with ordinary acupuncture technique, and the acupuncture treatment group was treated with Zhulian stimulant type Ⅰ acupuncture, 1 time/d, 30 minutes/time. Samples were taken after 4 weeks of treatment. The maximum bladder volume, residual urine volume and wet weight of the bladder were detected. The morphology of rat bladder was observed by HE staining. The expression level of BDNF was detected by immunohistochemistry. The expression of cAMP was detected by Western blot. The level of TrkB was determined by ELISA. Real-time fluorescence quantitative PCR (RT-PCR) was used to detect the mRNA expressions of BDNF and cAMP.Results:Compared with model group, maximum bladder volume, residual urine volume and wet weight of bladder in Western medicine group, ordinary acupuncture group and acupuncture treatment group decreased ( P<0.01), and those in Western medicine group and acupuncture treatment group were lower than those in ordinary acupuncture group ( P<0.01). The expressions of BDNF mRNA and protein, cAMP mRNA and protein in Western medicine group, ordinary acupuncture group and acupuncture treatment group increased ( P<0.05), and the level of TrkB increased, and the Western medicine group and acupuncture treatment group were higher than that in ordinary acupuncture group ( P<0.05). Conclusions:Zhuliping stimulant type Ⅰ acupuncture has a protective effect on the bladder function of diabetic rats. The mechanism may be related to the up-regulation of BDNF and mRNA, TrkB, cAMP and mRNA expressions.

Effects of acupuncture combined with Buyang Huanwu Decoction on the cerebral blood flow hypo perfusion model rats with carotid artery stenosis based on high-throughput sequencing / 国际中医中药杂志

Objective:To explore the effects of acupuncture combined with Buyang Huanwu Decoction on intestinal flora in cerebral blood flow hypo perfusion model rats with carotid artery stenosis.Methods:Totally 40 rats were randomly divided into sham-operation group, model group, TCM treatment group and acupuncture and drug combination treatment group, with 10 rats in each group. Except the sham-operation group, the other groups were prepared cerebral ischemia model by needle control and thread embolism method. TCM treatment group received Buyang Huanwu Decoction 100 mg/kg for gavage, once a day, and the intervention lasted for 2 weeks. In the acupuncture and drug combination group, based on the TCM treatment group, Baihui and its left and right sides of 2 mm were selected for acupuncture, once a day, and continuous intervention was performed for 2 weeks. Neurological function evaluation and behavioral function score were performed 7 and 14 days after administration, respectively. 16S rRNA sequencing was used to comprehensively characterize the structure and composition of fecal microflora of rats in each group. Linear discriminant analysis Effect Size (LEfSe) was used to analyze the difference of intestinal bacteria among groups.Result:On the 7th and 14th day after administration, compared with the model group, the neurological function score in the TCM treatment group and the acupuncture and drug combination group decreased ( P<0.05), and the behavioral function score increased ( P<0.05). Compared with model group, the Shannon index of TCM treatment group and acupuncture and drug combination group increased ( P<0.05). The abundance of Firmicutes increased ( P<0.05), and the abundance of Bacteroidetes and Proteobacteria decreased ( P<0.05); the abundance of Clostridia increased ( P<0.05), and the abundance of Gammaproteobacteria decreased ( P<0.05). The abundance of Escherichia-Shigella and Bacteroides decreased ( P<0.05); the abundance of lactobacillus significantly increased ( P<0.05). Conclusion:Acupuncture combined with Buyang Huanwu Decoction can improve the symptoms of cerebral hypoperfusion model rats with carotid artery stenosis, and the mechanism may be to increase the abundance of probiotics.