Protective effect of NeuroEPO on the reproduction of diabetic rats / Efecto protector de la NeuroEPO en la reproducción de ratas diabéticas

Introduction: Pregestational diabetes constitutes a reproductive risk which requires new treatment strategies. NeuroEPO, a variant of the recombinant human erythropoietin produced in Cuba, has neuroprotective and hypoglycemic effects which can be considered for the treatment of this entity. Objective: To evaluate the protective effect of NeuroEPO on the reproduction of diabetic rats. Material and Methods: Four groups of adult female Wistar rats with streptozotocin-induced diabetes were used. During pregnancy, one group received the vehicle and the rest of the groups received different doses of NeuroEPO (0,5 mg/kg, 0,75 mg/kg, and 1 mg/kg) subcutaneously, on alternate days, for a total of six applications. A group of non-diabetic rats was used as a control group. Glycemia and reproductive variables were evaluated. For comparisons, Analysis of Variance and Fisher's Exact Test were used. There were significant differences with p-values less than 0,05. Results: The group with vehicle presented maintained hyperglycemia, fewer implantations, and embryos, and increased gestational losses. In the group receiving 0,5 mg/kg of NeuroEPO, glycemia decreased significantly and the results of the reproductive variables were similar to the group of non-diabetic rats. With higher doses of NeuroEPO, gestational losses were increased. No congenital malformations were identified in either group. Conclusions: The repeated administration of 0,5 mg/kg of NeuroEPO has a beneficial effect on the reproduction of diabetic rats, which may be associated with the reduction of hyperglycemia. Other cytoprotective mechanisms of NeuroEPO should be evaluated in future studies(AU)

Introducción: la diabetes pre-gestacional constituye un riesgo reproductivo, lo que requiere nuevas estrategias de tratamiento. Teniendo en cuenta que la NeuroEPO, una variante de la eritropoyetina recombinante humana producida en Cuba, tiene efectos neuroprotectores e hipoglicemiantes. Objetivo: evaluar el efecto protector de la NeuroEPO en la reproducción de ratas diabéticas. Material y Métodos: se utilizaron cuatro grupos de ratas Wistar hembras adultas, con diabetes inducida por estreptozotocina. Durante la gestación, un grupo recibió el vehículo y el resto diferentes dosis de NeuroEPO (0,5 mg/kg, 0,75 mg/kg y 1 mg/kg), por vía subcutánea, en días alternos, para un total de seis aplicaciones. Se empleó un grupo de ratas no-diabéticas como control. Se evaluó la glicemia y variables reproductivas. Para las comparaciones se empleó el Análisis de Varianza y la Prueba Exacta de Fisher. Las diferencias se consideraron significativas con valores de p menores que 0,05. Resultados: el grupo con vehículo presentó hiperglicemia mantenida, menor número de implantaciones y embriones, e incremento de las pérdidas gestacionales. En el grupo que recibió 0,5 mg/kg de NeuroEPO, la glicemia disminuyó de forma significativa y los resultados de las variables reproductivas fueron similares al grupo de ratas no-diabéticas. Con las dosis superiores de NeuroEPO se incrementaron las pérdidas gestacionales. No se identificaron malformaciones congénitas en ninguno de los grupos. Conclusiones: la administración reiterada de 0,5 mg/kg de NeuroEPO tiene efecto beneficioso en la reproducción de ratas diabéticas, que puede estar asociado a la reducción de la hiperglicemia. Otros mecanismos citoprotectores de la NeuroEPO deben ser evaluados en futuros estudios(AU)

Mechanism of hepatotoxicity induced by ethanol extract of Dysosma versipellis based on "quantity-weight-evidence" network toxicology / 中国中药杂志

In this study, the toxicological/pharmacological research method of "quantity-weight-evidence" network was first proposed and practiced to supplement the existing methodology of network toxicology. We transformed the traditional qualitative network into a quantitative network in this study by attributing weights to toxic component content and target frequency, which improved the reliability of data and provided a research idea for the systematic safety evaluation and toxicological research of Chinese medicinal herbs. Firstly, 50% ethanol extract of Dysosma versipellis(DV) was administrated to rats via gavage and the potential hepatotoxic components were identified by serum pharmacochemistry. Then, the component targets were obtained from SwissTargetPrediction, PharmMapper and other online databases, and the target weights were given according to the relative content of components and target fishing frequency. Meanwhile, the targets of hepatotoxicity were predicted from online databases such as Comparative Toxicology Database(CTD) and GeneCards. Subsequently, protein-protein interaction analysis and KEGG pathway enrichment were performed with the STRING database. Finally, the quantitative network of "toxic components-weighted targets-pathways" was constructed. Eleven potential toxic compounds were predicted, including podophyllotoxin, podophyllotoxone, deoxypodophyllotoxin, and 6-methoxypodophyllotoxin. A total of 106 hepatotoxic targets and 65 weighted targets(e.g., Cdk2, Egfr, and Cyp2 c9) were identified. The results of Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment showed that these targets could act on PI3 K-AKT, MAPK, and Ras signaling pathways to play a role in inflammatory response and oxidative stress. However, traditional network toxicology showed that 51 targets such as AKT1, Alb, and Stat3 may lead to hepatotoxicity by mediating inflammation and cell proliferation. In conclusion, we proposed "quantity-weight-evidence" network toxicology in this study and used it to study the mechanism of DV-induced hepatotoxicity in rats. This study confirms the feasibility of this new methodology in toxicological evaluation and further improves the systematic evaluation of the safety of Chinese medicinal herbs.

In vitro inhibitory effects of acteoside on 6 kinds of CYP enzymes in liver microsomes of rats / 中国药房

OBJECTIVE To inv estigate the in vitro inhibitory effects of acteoside on cytochrome P 450（CYP）enzymes in liver microsomes of rats. METHODS Using probe substrates method ，acteoside（0.1，0.3，1，3，10，30 μmol/L）was incubated with probe substrates phenacetin ，mephentoin，diclofenac，coumarin，dextromethorphan and testosterone （substrates of CYP 1A2， CYP2C19，CYP2C9，CYP2A6，CYP2D6 and CYP 3A4 enzymes，respectively）in liver microsomes of rats. Another blank control group and positive inhibitor group [ α-naphthoflavone，ticlopidine，sulfabendazole，pilocarpine，quinidine and ketoconazole （inhibitors of CYP 1A2，CYP2C19，CYP2C9，CYP2A6，CYP2D6 and CYP 3A4 enzymes，respectively）] were set up. Using indapamide as the internal standard ， the contents of corresponding metabolites （acetaminophen， 4-hydroxymephenytoin， 4-hydroxydiclofenac，7-hydroxycoumarin，dextran，6 β-hydroxytestosterone） were detected by ultra high performance liquid chromatography-tandem mass spectrometry . The IC 50 values were calculated by GraphPad v 8.0 software. By computer molecular docking technology ，acteoside and positive inhibitors were molecularly docked with the CYP enzyme ，and the binding mode and strength of the two molecules were analyzed. RESULTS The IC 50 values of acteoside to CYP 1A2 and CYP 2A6 enzymes were more than 30 μmol/L，and those of acteoside to CYP 2D6，CYP2C19，CYP2C9 and CYP 3A4 enzymes were 24.87，21.52，12.56 and 7.55 μmol/L，respectively. The hydrogen bond and hydrophobic force could form between acteoside and CYP 3A4 enzyme，and the hydrogen bond and electrostatic interaction could form between ketoconazole and CYP 3A4 enzyme. The binding free energy of acteoside and ketoconazole to CYP 3A4 enzyme were － 10.2 and － 12.4 kcal/mol （1 kcal/mol＝4.19 kJ），respectively. CONCLUSIONS Acteoside shows moderate inhibitory effect on CYP 3A4 enzyme in liver microsomes of rats ，and its affinity is equivalent to that of positive inhibitor ；the compound shows weak inhibitory effect on other 5 CYP enzymes.

Genotoxicity assessment of three reference chemicals in rat hepatocytes by alkaline comet assay / 上海预防医学

Objective:To establish a comet test method for detection of genotoxicity of three reference chemicals in rat liver cells. Methods:6-10 week old Sprague Dawley rats were randomly divided into 4 groups， with normal saline （0.9% NaCl solution） as negative control group. Animals in three test groups were treated， respectively， with ethyl methanesulfonate （EMS） 200 mg/kg， N-methyl-N-nitrosourea （MNU） 50 mg/kg， and D-mannitol 2 000 mg/kg. There were 10 animals in each group， 5 males and 5 females. The animals received two times （21 h interval） of test compounds through intragastric administration， and their clinical symptoms and body weight changes were recorded during the experiment. The rats were sacrificed 3 h after the last exposure. The liver was weighed， then used to prepare single-cell suspensions for the alkaline comet test which determines the average tail DNA content percentage （DNA%） of hepatocytes and other comet indicators. Results:（1） D-mannitol， EMS and MNU did not show significant toxicity in the whole animal. （2） The mean values of tail DNA content percentage （DNA%） of rat hepatocytes in EMS ［（60.07±24.69）%］ and MNU ［（41.66±22.35）%］ groups were higher than that in the negative control group ［（2.32±1.39）%］ and the difference was statistically significant （P<0.05）. The difference between D-mannitol group ［（3.06±3.30）%］ and the negative control group was not significant （P>0.05）. Conclusion:This laboratory has established a comet test method using hepatocytes from treated rats. Among three testing chemicals， EMS and MNU have displayed genotoxicity by this assay， but no genotoxicity was observed in D-mannitol treated animals.

Bioavailability Study Of Ondansetron Gel In Rabbits And Human Volunteers Appling Uplc As Analytical Tool And Evaluation Of The Antiemetic Effect Of Ondansetron Gel In Cisplatin-Induced Emesis In Rats

Objective: This study was undertaken to determine the bioavailability of ondansetron gel in experimental animals and humans applying UPLC as an analytical tool and evaluation of the antiemetic effect of ondansetron gel in cisplatin-induced emesis in rats. Methods: Ondansetron gel (F13: sodium alginate 7% w/w) was used, marketed I. V. ondansetron (Zofran) ® was chosen as reference. The bioavailability study in rabbits was selected as a parallel design using nine healthy rabbits divided into three groups whereas, bioavailability study in humans was an open-label, wherein 6 healthy subjects administered ondansetron gel. The potential effect of ondansetron gel was evaluated for the prevention of different phases of emesis motivated by exposure to antineoplastic drugs (cisplatin) by determination of body weight loss, water and food intake applying kaolin-pica model in rats using seventy-two rats divided into six groups. Results: Ondansetron gel (0.5%) showed detectable plasma concentration 22.833±2.17 ng/m1 after ¼ h and 419.55±2.17 ng/ml after 1-h post-treatment in rabbits and human respectively and concentration was maintained above-reported minimum effective concentration for more than 2.5 h for rabbits and 7 h for humans compared to 1.75 h after I. V. administration. The ondansetron gel significantly reduces all phases of cisplatin-induced emesis and a decrease in body weight, water, and food consumption was significantly attenuated. Conclusion: Based on the high efficacy of gel on emesis induced by cisplatin, and its high bioavailability, transdermal ondansetron gel could be a promising convenient system to prevent nausea and vomiting following administration of antineoplastic drugs.

Effects of different swimming intensities on the bone properties of the tibia and femur of Wistar Rats in which knee rheumatoid arthritis was induced / Efectos de las diferentes intensidades de la natación en las propiedades óseas de la tibia y fémur de Ratas Wistar con artritis reumatoide inducida en rodillas

Rheumatoid arthritis (RA) is a disease triggered by environmental and genetic factors. Research suggests that physical exercise has benefits such as delaying functional disability. In vivo studies using experimental models of arthritis can provide useful information about these benefits. to analyze the effects that different intensities of aquatic physical exercise have on the proprieties of the bones in induced arthritis in knees of Wistar rats. Male Wistar adults rats (n=20) were divided into 5 groups: Group Control Arthritis (GCA) n=4, Group control Placebo (GCP) n=4, Group Low Physical Activity (GB) n=4, Group Moderate Physical Activity (GM) n=4 and Group Intense Physical Activity (GI) n=4. The physical activity groups got an intra-articular injection of Zymosam on the right knee; the GCA received saline solution in the right knee; the GCP was submitted to the stress of the needle. The animals were submitted to aquatic activity for 30 minutes, 4 times a week for 5 weeks, and the intensity of the exercise was determined by a weight placed on their back: GB=1 %, GM=5 %, GI=15 % of their body weight. It was observed that the group GB, and the groups that did not exercise GCA and GCP, gained more weight compared to the group GM. In relation to the bone mineral content of the tibia, there was a decrease in the GM group when compared to the GCP group, whereas in the tibial bone mineral density there was a decrease in the GM group compared to the GCP, GCA, GB. As for the area of the femur, the GI group presented an increase of it compared to the GB and GM groups. It is concluded that the high intensity exercises promote better results in bone properties.

La investigación sugiere que el ejercicio físico tiene beneficios como retrasar la discapacidad funcional de la artritis reumatoide. Los estudios in vivo que utilizan modelos experimentales de artritis pueden proporcionar información útil sobre estos beneficios. Se analizaron los efectos de las intensidades del ejercicio físico acuático sobre las propiedades de los huesos, en la artritis inducida en las rodillas de ratas Wistar. Las ratas Wistar macho adultas (n = 20) se dividieron en 5 grupos: grupo de control artritis (ACG) n = 4, grupo control placebo (CGP) n = 4, grupo de actividad física baja (GB) n = 4, grupo de actividad física moderada (GM) n = 4 y grupo de actividad física intensa (GI) n = 4. Los grupos de actividad física recibieron una inyección intraarticular de Zymosam en la rodilla derecha; el GCA recibió solución salina en la rodilla derecha; el CGP fue sometido a la tensión de una aguja. Los animales fueron sometidos a actividad acuática durante 30 minutos, 4 veces a la semana durante 5 semanas, y la intensidad del ejercicio se determinó mediante un peso colocado sobre su espalda: GB = 1 %, GM = 5 %, GI = 15 % de su peso corporal. Se observó que el grupo GB, y los grupos que no ejercitaron GCA y CGP, ganaron más peso en comparación con el grupo GM. En relación con el contenido mineral óseo de la tibia, hubo una disminución en el grupo GM en comparación con el grupo GCP, mientras que en la densidad mineral del hueso tibial hubo una disminución en el grupo GM en comparación con el GCP, GCA, GB. En cuanto al área del fémur, el grupo GI presentó un aumento en comparación con los grupos GB y GM. En conclusión el ejercicio de alta intensidad promueve mejores resultados en las propiedades óseas.

Rapid bioluminescence assay for monitoring rat CES1 activity and its alteration by traditional Chinese medicines / 药物分析学报

In traditional Chinese medicine herbs (TCM), including Radix Salviae Miltiorrhizae (Danshen), Radix Puerariae Lobatae (Gegen), Radix Angelicae Sinensis (Danggui), and Rhizoma Chuanxiong (Chuanxiong) are widely used for the prevention and treatment of cardiovascular diseases and also often co-administered with Western drugs as a part of integrative medicine practice. Carboxylesterase 1 (CES1) plays a pivotal role in the metabolisms of pro-drugs. Since (S)-2-(2-(6-dimethylamino)-benzothiazole)-4,5-dihydro-thiazole-4-carboxylate (NLMe) has recently been identified by us as a selective CES1 bioluminescent sensor, we developed a rapid method using this substrate for the direct measurement of CES1 activity in rats. This bioluminescence assay was applied to determine CES1 activity in rat tissues after a two-week oral administration of each of the four herbs noted above. The results demonstrated the presence of CES1 enzyme in rat blood and all tested tissues with much higher enzyme activity in the blood, liver, kidney and heart than that in the small intestine, spleen, lung, pancreas, brain and stomach. In addition, the four herbs showed tissue-specific effects on rat CES1 expression. Based on the CES1 biodistribution and its changes after treatment in rats, the possibility that Danshen, Gegen and Danggui might alter CES1 ac-tivities in human blood and kidney should be considered. In summary, a selective and sensitive biolu-minescence assay was developed to rapidly evaluate CES1 activity and the effects of orally administered TCMs in rats.

Effects of Chelidonine on Proliferation ，Collagen Synthesis and TGF-β1 Receptor of Activated Hepatic Stellate Cells CFSC- 8B in Rats / 中国药房

OBJECTIVE： To investigate the effects of chelidonine on proliferation， collagen synthesis and TGF-β1 receptor of activated hepatic stellate cells CFSC-8B. METHODS： CFSC-8B cells in logarithmic phase were collected and then divided into normal control group， model group， solvent group （ethanol）， positive control group （1 μg/mL colchicine ethanol solution）， chelidonine low， medium and high concentration groups （2.1， 4.2， 8.4 μg/mL chelidonine ethanol solution）. Except for normal control group， other groups were activated with 20 μg/L TGF-β1 for 24 h； the latter 5 groups were intervened with relevant medicine for 24 h. Cell proliferation of activated cells was assayed by CCK-8 assay. Hydroxyprolin （Hyp） content was assayed by enzyme digestion； the levels of typeⅠ collagen （Col-Ⅰ） and type Ⅲ collagen （Col-Ⅲ） were assayed by ELISA； the expressions of TβR-Ⅰ and TβR-Ⅱ protein were assessed by Western blot； mRNA expressions of α-SMA， TβR-Ⅰ and TβR-Ⅱ in hepatic stellate cells were assessed by RT-PCR. RESULTS： Compared with normal control group， cell proliferation rate， Hyp content， the levels of Col-Ⅰ and Col-Ⅲ， the protein expressions of TβR-Ⅰ and TβR-Ⅱ as well as mRNA expressions of α-SMA， TβR-Ⅰ and TβR-Ⅱ were increased significantly （P＜0.05）. Compared with model group， there were no significant difference in above indexes of hepatic stellate cells in solvent group （P＞0.05）； there were no significant difference in the proliferation rate of hepatic stellate cells in chelidonine low concentration group （P＞0.05）， above indexes of hepatic stellate cells were decreased significantly in positive control group and chelidonine high concentration group （P＜0.05）. The decrease of Hyp and Col-Ⅲ levels were not significant in chelidonine medium concentration， but other above indexes were decreased significantly （P＜0.05）. Compared with chelidonine medium concentration group， the rate of cell proliferation， Col-Ⅰ level， protein and mRNA expressions of TβR-Ⅰ and TβR-Ⅱ were decreased significantly in chelidonine high concentration group （P＜0.05）. CONCLUSIONS： Chelido- nine can inhibit the proliferation， collagen synthesis as well as the protein and mRNA expressions of TβR-Ⅰand TβR-Ⅱ in activated CFSC-8B cells.

Screen absorption enhancer for intranasal administration preparations of paeoniflorin based on nasal perfusion method in rats / 中国中药杂志

This experiment was aimed to screen the absorption enhancer for intranasal administration preparations of paeoniflorin. In this study, HPLC method for determination of paeoniflorin in perfusion liquid was established and the improved model of nasal perfusion in rats was used to screen out the species and amounts of absorption enhancer. In order to avoid the influence of the secretion and absorption of nasal cavity on the volume of perfusion fluid, the residual dose was calculated by using the volume correction method. Linear regression was carried out between the logarithm to the percentage of the residual dose and the corresponding time, and the slope of the regression line was exactly the absorption rate constant. Experimental results showed that hydroxypropyl-β-cyclodextrin and water-soluble azone can significantly improve the nasal absorption of paeoniflorin. Furthermore, water-soluble azone had the highest absorption rate constant and the best promoting penetration effect on intranasal administration preparations of paeoniflorin. It was also found that when the mass concentration of water-soluble azone in the perfusion liquid increased from 5 g•L⁻¹ to 20 g•L⁻¹, the absorption rate constant was gradually increased and peaked at 20 g•L⁻¹. When the mass concentration was increased to 30 g•L⁻¹, the absorption rate constant was decreased, indicating that the best mass concentration of water-soluble azone was 20 g•L⁻¹.

The influence of growth factors on skin wound healing in rats / A influência de fatores de crescimento na cicatrização de feridas cutâneas de ratas

ABSTRACT INTRODUCTION: Healing is a process that restores the physical integrity of body structures. It is a dynamic, complex, multicellular process that involves the extracellular matrix, cytokines, blood cells, and growth factors. Growth factors are proteins that activate and stimulate cell proliferation through the activation of angiogenesis, mitogenesis, and gene transcription, accelerating the healing process. OBJECTIVE: To assess the influence of growth factors on the healing process of wounds made on the backs of female rats compared to the control wound, through macro and microscopy. METHODS: This study used 45 female Wistar rats, in which three wounds were made on the back. The first was the control wound, the second received epithelial growth factor injection, and the third received a combination of factors. Macroscopic and microscopic assessments were performed on the third, seventh, and 15th days of the experiment. For microscopic analysis, hematoxylin-eosin staining was utilized to assess the inflammatory process; vimentin, for assessment of blood vessels and fibroblasts, and Sirius Red for collagen assessment. RESULTS: In the macroscopic assessment, the use of growth factors resulted in faster healing and decrease of granulation tissue on days seven and 15; (80.31% reduction in the control wound vs. 83.24% in the epithelial wound vs. 100% in the mixed wound). Utilizing microscopy, at the three stages of the experiment, there were no significant differences between the three wounds; however, when comparing the day of euthanization for each type of wound, there was a favorable outcome for epithelial and mixed wounds (between the third vs. 15th day, p < 0.001, and in the comparison of the seventh vs. 15th day; p = 0.002 and p = 0.001 for epithelial and mixed wounds, respectively) with a higher number of fibroblasts, angiogenesis, and collagen type I. CONCLUSION: The use of growth factors accelerates healing, stimulates greater angiogenic activity, and accelerates fibroplasia and collagen maturation.

Resumo Introdução: A cicatrização é um processo de restauração da integridade física das estruturas do corpo. É um processo dinâmico, complexo, multicelular que envolve matriz extracelular, citosinas, células sanguíneas e fatores de crescimento. Os fatores de crescimento são proteínas que estimulam e ativam a proliferação celular mediante a ativação da angiogênese, mitogênese, transcrição genética, acelerando o processo de cicatrização. Objetivo: Avaliar a influência dos fatores de crescimento no processo cicatricial de feridas realizadas no dorso de ratas em comparação com a ferida, controle através da macro e microscopia. Método: Foram utilizadas 45 ratas Wistar, submetidas à criação de três feridas no dorso. A primeira controle a segunda com injeção de fator de crescimento epitelial e a terceira com fator misto. As avaliações macroscópicas e microscópicas foram realizadas no 3º, no 7º e no 15º dia do experimento. Para análise microscópica, utilizou-se coloração de Hematoxilina-Eosina para avaliar o processo inflamatório; vimentina, para a avaliação dos vasos e fibroblastos, e Sirius Red, para avaliar o colágeno. Resultados: Na avaliação macroscópica, o uso de fatores de crescimento proporcionou cicatrização mais rápida e diminuição do tecido de granulação no 7º e 15º dia (80,31% de redução na ferida controle vs. 83,24% na ferida epitelial vs. 100% na ferida mista). Na microscopia, nos três momentos do experimento, não foram encontradas diferenças significativas entre as três feridas; entretanto, quando comparados os dias de morte em relação a cada tipo de ferida, observou-se resultado favorável para as feridas epiteliais e mistas (entre 3º × 15º dia apresentou p < 0,001 e na comparação entre 7º × 15º dias; p = 0,002 e p = 0,001 para as feridas epiteliais e mistas) com maior número de fibroblasto, angiogênese e colágeno tipo 1. Conclusão: A utilização de fatores de crescimento acelera a cicatrização, estimula maior atividade angiogênica, acelera a fibroplasia e maturação do colágeno.

Effect of salvianolic acid B on nasal absorption in rats in situ / 中国中药杂志

This experiment focused on the effect of salvianolic acid B's nasal absorption characteristics in rats. In the study, HPLC determination of salvianolic acid B(SalB) in perfusion liquid was established to examine the SalB nasal irritation in different pH buffers and stability in nasal perfusion solution, and systematically study in vivo nasal absorption characteristics of SalB. Improved rats were adopted to establish the in situ nasal perfusion model to measure the release of total protein and lactate dehydrogenase in perfusion fluid, quantitatively evaluate the nasal irritation and the stability in perfusion liquid of pH 4.0, 5.0, 6.0 SalB phosphate buffer, compare the absorption of SalB in pH 5.0 buffer solution with low, medium and high concentrations (200, 400, 800 mg•L⁻¹). According to the results, nasal irritation: pH 4.0>pH 5.0>pH 6.0, RSD of pH 6.0 SalB buffer solution within 24 h was 3.1%, stability was poor. PH 5.0 SalB buffer solution had a smaller irritation and good stability. According to the nose perfusion test in rats, the nasal absorption of SalB fitted the first-order process and could be considered as passive absorption based on concentration gradient. SalB buffer solution of pH 5.0 had also a small nasal irritation and good stability, with a good absorption in rat nasal perfusion test, which therefore had a certain significance for the development of SalB nasal formulation.

Influence ofZhulian acupuncture exciting method on hypoxic ischemic brain damage in rats to oxidative stress / 国际中医中药杂志

Objective To observate theZhulian acupuncture exciting method in different time of hypoxia ischemia brain injury in rat brain tissue malondialdehyde (MDA), monoamine oxidase (MAO), nitric oxide (NO) and glutathione peroxidase (GSH-PX) content.Methods 7 days old rats were randomly divided into a excitation method acupuncture group I, a excitation method acupuncture group II, a model group, a sham operation group, a normal control group, 10 rats in each group. Excitation method acupuncture group I and normal control group were given stimulation ofZhulian acupuncture exciting method one type technique from 24 h model manipulation, excitation method acupuncture II group from the beginning of the eighth day given Stimulation ofZhulian acupuncture exciting method one type technique. The sham operation group and the model group were not treated by acupuncture. The animals were sacrificed at the twenty-first day after making the model, determined brain tissue MDA、MAO、NO and GSH-PX Content.Results Compared with the model group, MDA (3.4 ± 0.87 nmol/mgvs. 5.50 ± 1.58 nmol/mg) content decreased in the excitation method acupuncture group I (P<0.05). The NO (12.43 ± 3.47μmol/mgvs. 17.10 ± 5.82μmol/mg) content decreased in the excitation method acupuncture group II (P<0.05). MAO (32.12 ± 11.15 U/mg, 31.01 ± 9.92 U/mgvs. 40.90 ± 11.02 U/mg) content were decreased in both excitation method acupuncture group I and group II (P<0.05), while the GSH-PX (2.61 ± 1.20 U/mg, 2.61 ± 1.37 U/mgvs. 1.43 ± 0.49 U/mg) content were increased (P<0.01). ConclusionZhulian acupuncture exciting method one type technique can decrease the content of MDA, MAO and NO reduce the content of hypoxic-ischemic brain injury in rat brain tissue, increase the content of GSH-PX, promote the removal of immature rats with hypoxic ischemic brain damage brain tissue metabolism, and protect brain function.

PHARMACOLOGICAL EFFECTS OF SILLICHOL ON BILE SECRETION AND / Монголын эм зүй, эм судлал

Varieties of plants and lyophilized bovine bile have been used for increase secretion of bile in traditional systems of medicine of various countries. Following many articles note on the benefi cial effects of lyophilized bovine bile particularly on the wound healing and gastric protection effects, there is paucity of reports in literature on its effects on a bile secretion, a bile bilirubin, bile cholesterol and a plasma cholesterol levels. Sillichol contains lyophilizedbovine bile, liver hydrolisate, yarrow extract and silymarin. The aim of this study was to find out the effect of bile fl ow, bile bilirubin concentration bile cholesterol level and hepatoprotective of Sillichol. Sixteen adult male wistar rats (weighing between 200-250 gr) were used in the study. They were randomly assigned into control and sillichol group comprising 4 in each group. Thereafter, they were weighed and anaesthetized with ketamine (2ml/200gr body weight) muscle leg and quickly pinned to a dissecting board. Laparotomy was performed and liver lobes were defl ected anterolaterally to expose the common bile duct. The common bile duct was cannulated with a portex cannula (0.5 mm diameter) after a semitransection was made on the bile duct. The cannuls was held in place with thread tied over it and around the bile duct.The bile was collected for 8 hours from each rat studied according to method of Rozuet Jousse. The rate of bile fl ow was noted, the volume of bilirubin, bile cholesterol levels were determined in the control and test groups. Moreover, total of 18 wistar rats (200-250 gr) were obtainedfrom laboratory house of Drug research institute and acclimated for 10 days before starting the experiment. Liver toxicity was induced by the subcutaneous injection of carbon tetrachloride (CCL4, 0.4 ml/100gr), 1:1 diluted with paraffi n oil, for four successive days of the experiment (N.P.Scakun et al, 1983). The rats were divided randomly into 3 groups comprising 6 rats in each group and fed the same diet throughout the experimental period. Mean values of bile cholesterol and bilirubinlevels, rate of bile secretion in the control and sillichol group. Bile cholesterol levels were signifi cantly decreased in the sillichol group compared with the control group (60.3±0.88 mg/dl vs 62.6±1.21mg/dl, p<0.05). Rate of bile secretion was signifi cantly increased in the experimental compared with the control group(10.21±0.25 ml/8hr vs 4.18±0.25 ml/8hr, p<0.05). Total bilirubin, conjugated and unconjugatedbilirubin concentrations in both sillichol and control groups were not signifi cantly different (p<0.01). The activities of GOT, GPT and ALP were estimated in serum samples as the liver function biomarkers using biochemical diagnostic test. The CCL4 treatment markedly affected the liverspecifi c enzymes. It was found that a signifi cant (p<0.05) increase in serum GOT, GPT and ALP activities of CCL4 treated rats. After the treats, hepatic biomarkers were elevated in the serum due to release of the enzymes from damaged liver. GOT (69.8±1.5), GPT (103.9±1.2), ALP (23.8±0.2) and Cholesterol (67.7±13.6) andtriglyceride (64.0±3.3) levels weredecreasedsignifi cantly (p<0.05) in the sillichol groupcompared with the control group. Silichol is decreasing concentration of cholesterol and bilirubin’s level in bile, constantly after administration of drug. Also, liverpreparation is increasing bile acid secretion in hepatocytes and a speed of secretion.From the results of pharmacological study concluded that involves CCL4 induced acute toxic hepatitis, liver preparation has hepatoprotective effect by protecting the liver cells from injury, improving the regeneration process and by correcting metabolic functions of the liver.

Determination of Pharmacokinetics Differences of Ammuxetine Isomers in Rat Plasma Using On-Line Solid Phase Extraction with Liquid Chrommatography-Tandem Mass Spectrometry / 分析化学

An on-line solid phase extraction ( SPE ) coupled with HPLC-MS/MS method was developed to determine S-ammuxetine and R-ammuxetine in rat plasma. The sample preparation consisted of the following steps:A protein precipitation extraction used methanol and acetonitrile ( 50:50 , V/V ); an on-line SPE treatment to remove most matrixes in plasma;an enrichment and separation step used a C18 analytical column. S-and R-ammuxetine were determined by tandem mass spectrometry. The SPE column was a Retain PEP Javelin (10 mm × 2. 1 mm × 5 μm), while the chromatographic separation was achieved using a ZORBAX SB-C18 (50 mm × 2. 1 mm × 3. 5 μm) analytical column with an isocratic mobile phase composed of acetonitrile-water-formic acid (40:60:0. 1, V/V/V, 0. 3 mL/min). The selected reaction monitoring mode of the positive ion was performed and the precursor to the product ion transitions of m/z 292 . 1/154 . 0 and m/z 260. 4/116. 2 were used to measure S-ammuxetine, R-ammuxetine and internal standard (propranolol). The method was linear over a concentration range from 0 . 2 to 1000 μg/L with the correlation coefficients of 0 . 9903 and 0 . 9951 . The average intra-day precision values were 1 . 2% -12 . 0% for S-ammuxetine and 0. 4%-11. 2% for R-ammuxetine, respectively. The average recoveries were 94. 2%-101. 6% for S-ammuxetine and 94. 3% -109. 4% for R-ammuxetine. Compared to the literature, the sensitivity of this method increased dramatically. The present method has been successfully applied to the preclinical rat research of ammuxetine isomers following intragastric administration.

Estudio preliminar de la miocarditis chagásica aguda experimental y su relación con la administración de esteroides sexuales / Preliminary study of experimental acute Chagasic cardiomyopathy and its relation to sex steroid administration / Estudo preliminar de miocardite chagásica aguda experimental e sua relação com a administração de esteróides sexuais

Introducción. En nuestro modelo experimental, la infección aguda por Trypanosoma cruzi (T. cruzi) en ratas adultas cursa con una parasitemia poco evidente, mientras que en los animales prepúberes (PP) se registran parasitemias más elevadas. Estas discrepancias podrían asociarse a la inmadurez inmunológica que exhiben los animales más jóvenes, pero asimismo, a la diferente madurez sexual del huésped al momento de la infección. Siendo la testosterona (T) una hormona capaz de influir sobre las células del sistema inmune y en consecuencia modificar el curso de las infecciones parasitarias, nos propusimos evaluar el efecto de dosis fisiológicas de T sobre la miocarditis aguda en fase temprana en ratas PP. Material y métodos. Se inocularon 1 millón de T. cruzi al destete y dos dosis de T de 1 mg/kg de peso, previo al T. cruzi. Se realizaron los controles respectivos, incluido un grupo experimental que recibió bicalutamida, antagonista de la T (5 mg/kg/día) +T+T. cruzi. Se evaluó parasitemia a los 7, 10 y 14 días post infección (pi) y se realizó el estudio anátomo-patológico de corazón, timo y bazo a los 4, 7 y 14 días pi. Resultados. A los 14 días pi, los animales que recibieron dosis fisiológicas de T presentaron un incremento significativo en la parasitemia y desarrollaron una mayor esplenomegalia que el resto de los grupos infectados. El estudio histológico de esos grupos reveló una miocarditis de intensidad similar -moderada a intensa- y nidos de amastigotes, mientras que en los animales sacrificados al día 4 y 7, se observó nidos de amastigotes sin reacción inflamatoria. Los controles no presentaron alteraciones histológicas. Conclusiones. La administración de T en los animales PP, previo a la infección con T. cruzi, propició la replicación temprana del parásito, evidenciada por el aumento en la parasitemia; sin embargo, no fue capaz de modificar la lesión cardíaca aguda en fase temprana ni tardía.

Background. According to our experimental model, along the acute phase of Chagas illness, adult rats infected with Trypanosome cruzi (T. cruzi) presents very low and almost undetectable parasitemias, whereas in prepubertal animals (PP) parasitemias reported were higher than in the adult ones. These differences could be associated with the immunological immaturity exhibited by younger animals, but also owing to the different sexual maturity of the host at the time of infection. As testosterone (T) is a hormone that can influence immune system cells and thus modify the course of parasitic infections, we have evaluated the effect of physiological doses of T on early acute stage of myocarditis in rats PP. Methods and material. Two doses of T (1 mg/kg) were inoculated to weaning rats (prior infection) followed by the inoculation of 1 million of T. cruzi trypomastigotes. The proper controls were performed, including an experimental group which received a concomitant therapy of bicalutamide, an antagonist of T (5 mg/kg/day), plus T and T. cruzi inoculation. Parasitemia was assessed at 7, 10 and 14 days post infection (pi) and the anatomopathological studies of heart, thymus and spleen were also performed at 4, 7 and 14 days pi. Results. At 14 day pi, those animals receiving physiological doses of T showed a significant increase in parasitemia and developed a higher splenomegaly compare to the rest of the infected groups. The histological examination of these groups presented a myocarditis of similar intensity -moderate to intense-, and amastigotes nests, while at days 4 and 7, amastigotes nests were observed without inflammatory reaction. Controls did not present histological alterations. Conclusions. Administration of T in the PP animals prior to T. cruzi infection led to an early parasite replication, as evidenced by the increase in parasitemia, however, it was not able to modify the acute cardiac injury during the early or late stage.

Introdução. Em nosso modelo experimental, a infecção aguda pelo Trypanosoma cruzi (T. cruzi) em ratos adultos cursos com parasitemia evidente pouco, enquanto que em animais pré-púberes (PP) são registrados parasitemias mais elevadas. Estas discrepâncias podem estar associadas à imaturidade imunológica exibidos por animais mais jovens, mas também para a diferente maturidade sexual do parasitado no momento da infecção. Como o hormônio testosterona (T) que pode influenciar as células do sistema imunológico e, assim, modificar o curso das infecções parasitárias, nós avaliamos o efeito de doses fisiológicas de T em estágio inicial miocardite aguda em ratos PP. Material e métodos. Um milhão de T. cruzi foram inoculados a desmame e duas doses de T de 1 mg / kg, antes a T. cruzi. Respectivos controles foram realizados, incluindo um grupo experimental que receberam bicalutamida, um antagonista da T (5 mg/kg/dia) + T + T. cruzi. Parasitemia foi avaliada aos 7, 10 e 14 dias após a infecção (ai) e realizado o estudo patológico do coração, timo e baço a 4, 7 e 14 dias ai. Resultados. Aos 14 dias ai, os animais que receberam doses fisiológicas de T mostraram um aumento significativo na parasitemia e desenvolveram uma maior esplenomegalia que outros grupos infectados. Exame histológico desses grupos revelou uma intensidade similar de miocardite -moderada a intensa-, e ninhos de amastigotas, enquanto em animais sacrificados nos dias 4 e 7, ninhos de amastigotas foram observados sem reação inflamatória. Os controles não apresentaram alterações histológicas. Conclusões. A administração de T nos animais PP antes da infecção com T. cruzi, levou à replicação inicial do parasita, como evidenciado pelo aumento da parasitemia, entretanto, não foi capaz de modificar a lesão cardíaca aguda na fase precoce ou tardia.

Effects of minocycline at subantimicrobial dosage in the treatment of experimental periodontitis in rats / 实用口腔医学杂志

Objective: To evaluate the effectiveness of minocycline at subantimicrobial dosage in the treatment of experimentally induced periodontitis in rats.Methods: Periodontitis was induced by silk ligature technique on 4 second molars in each of 16 adult male Sprague-Dawley rats and the animals were fed with 100 g/L sucrose drink. The rats were divided into 2 groups: (1)periodontitis group without treatment;(2)treatment group, the rats with periodontitis were treated by systemic administration of minocycline at 5 mg/(kg?d).Another 8 normal rats were used as the controls. Assessment was carried out at day 28 and 56 using a number of different visual, histological and ultrastructure approaches.Visual assessment included tooth mobility(TM), gingival index(GI), and alveolar bone loss. Histological examination included monocyte effusion,resorption lacunae with osteoclasts and percentage of the periodontal collagen.The collected data were statistically analyzed using variance test.Results: Systemic administration of minocycline at subantimicrobial dosage can significantly reduce GI, TM, resorption lacunae with osteoclasts and alveolar bone loss either at day 28 or at day 56; significantly inhibited monocyte effusion and the collagen degradation in the periodontium at day 56 in rats with periodontitis. Conclusion: Minocycline at subantimicrobial dosage may decrease alveolar bone loss and osteoclasts formation in periodontium with periodontitis.

Tratamento de ratos, experimentalmente infectados pelo Strongyloides venezuelensis, através da ivermectina administrada por via oral / The treatment of rats experimentally infected with Strongyloides venezuelensis by orally administered ivermectin

Em modelo experimental, baseado na infecção de ratos pelo Strongyloides venezuelensis, foi avaliada a atividade terapêutica de duas preparações de ivermectina, para usos veterinário e humano. Houve interesse em verificar a efetividade em relação a vermes adultos e formas larvárias. A administração dos fármacos ocorreu sempre por via oral e a posologia correspondeu à dose única de 0,2mg/kg. Considerados os vermes adultos e as formas larvárias, o produto para emprego veterinário propiciou eliminações expressas pelas porcentagens de 98,0% e 84,2%; quanto à outra preparação, as taxas situaram-se em 59,3% e 73,0%, respectivamente. O estudo revelou, então, utilidade do anti-helmíntico quando usada a via oral e, também, mostrou significativa ação sobre as formas larvárias, certamente valiosa quando vigente a modalidade disseminada da estrongiloidíase.

Strongyloides venezuelensis experimental infection in rats was treated by two different oral preparations of ivermectin, 0.2 mg/kg. One was a human formula used by WHO in the treatment of onchocerciasis; the other was a veterinary preparation. Adult worms and larvae were evaluated. The human formulation cleared both forms in 59.3% (adult worms) and 73.0% (larvae), whereas the veterinary one cleared 98.0% and 84.2%, respectively. The antilarval action is very useful when treating systemic strongyloidiasis.

Utilization of Supercompensated Glycogen of Hindlimb Muscles during Strenous Exercise in Rats / 영남의대학술지

The aim of the present investigation has been to evaluate the depletion pattern of the supercompensated glycogen of hindlimb muscles during strenous exercise in rats. The plan of the maximizing muscle glycogen stores is based on the fact that a glycogen-depleted muscle by exercise will have an increased avidity for glycogen when exposed to a high carbohydrate diet. The glycogen concentration of soleus, red gastrocnemius and plantaris muscle, and liver was measured at 0, 30 and 60 minutes during treadmill exercise. The experimental animals were divided into 5 group - Normal(N), Control(C), 1Hour(1HR:after 1hour of glucose ingestion), 2Hour(2HR:after 2hour of glucose ingestion) and Exercise-1Hour(EX-1HR:glucose ingestion after 1 hour of preloading treadmill exercise)group - for glycogen storage study. The glycogen concentration of soleus, red gastrocnemius and plantaris muscles in N group was 4.57+/-0.34, 5.11+/-0.24 and 6.55+/-0.20 mg/gm wet wt., respectively. The glycogen concentration of soleus and red gastrocnemius in EX-1HR group were about 1.9 and 1.8 times than that of N group, respectively, but the concentration of plantaris was not higher than that of N group. The glycogen concentration of liver in N group was 41.0+/-1.47mg/gm wet wt. and the concentration of the overnight fasted C group wad only 2.9% of the value of N group. The level of glycogen concentration of liver in the other glucose ingested groups(1HR, 2HR, including EX-1HR) was within 19 - 32% of that of N group. The blood glucose concentration of EX-1HR group was higher than that of N group, the plasma free fatty acid concentration of C and 2HR group was higher than that of N group, and the plasma insulin concentration of EX-1HR group was higher than that of N group. The concentration of supercompensated glycogen of soleus and red gastrocnemius were rapidly decreased during 30 minutes of exercise but there was almost no changes of the concentration during the other 30 minutes of continuing exercise. The concentration of N group during 30 minutes of exercise was decreased but more slowly than those of EX-1HR group. The remaining level of glycogen after 60 minutes of exercise in EX-1HR group was higher than that of N group. Taken together, the mobilization of endogenous muscle glycogen at the first stage of exercise was proportioned to the intial level of glycogen concentration, and later on, when exercise continued, the muscle glycogen level was stabilized. And the remaining level of supercompensated muscle glycogen after 60 minutes of exercise was higher than that of normally stored glycogen level. The mobilization of the glycogen stroed in slow and fast oxidative muscle fibers is faster than in the fast glycolytic muscle fibers during strenous exercise.