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Objective:To screen the active components, target genes and signaling pathways of Shaoteng Decoction in the treatment of Sjogren's syndrome by network pharmacology; To conduct relevant experimental verification to explore the mechanism of action of Shaoteng Decoction in the treatment of Sjogren's syndrome.Methods:The active components and targets of Shaoteng Decoction were collected by retrieving TCMSP. The target genes of Sjogren's syndrome were collected through the GeneCards database. The intersection targets of drugs and diseases were obtained by using Venn. The intersection targets were imported into the STRING database to obtain PPI networks, and the "drug-active component -therapeutic target-disease" network was constructed by Cytospace 3.7.2 software. The DAVID database was used for GO function enrichment analysis and KEGG pathway analysis. The 18 NOD mice were divided into model group, TCM group, hydroxychloroquine group, with 6 mice in each group, and 6 Balb/C mice were set as normal control group. TCM group was gavaged with 2.3 g/kg of Shaoteng Decoction, hydroxychloroquine group was gavaged with 60 mg/kg of hydroxychloroquine, and model group and normal control group were gavaged with equal volume of deionized water once a day for 4 consecutive weeks. The daily water intake of mice during the administration period was recorded, the pathological changes of submandibular gland tissue were observed by HE staining, and the levels of serum inflammatory factors IL-17 and TNF-α were determined by ELISA method.Results:39 main active components of Shaoteng Decoction, 1 062 targets of Sjogren's syndrome, and 64 targets of drug and disease intersection were obtained, including TNF, IL6, NCOA1, AKT1, TP53, etc. The treatment targets of Sjogren's syndrome mainly affected biological processes such as response to bacterium and cellular response to lipid, and regulated TNF-α pathway and IL-17 signaling pathway. The experimental results showed that the levels of TNF-α and IL-17 in the TCM group were lower than those in the model group ( P<0.05). Conclusion:Shaoteng Decoction can regulate IL-17 and TNF-α signaling pathways, inhibit inflammation, delay submandibular gland disruption, and alleviate the symptoms of Sjogren's syndrome.
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Objective We aimed to explore the therapeutic mechanism of electroacupuncture at Yintang(EX-HN3),Neiguan(PC6)and Zusanli(ST36)on the Hypothalamic-Pituitary-Adrenal axis(HPA)of functional dyspepsia(FD)rats.Methods Forty SD rats were randomly divided into blank group,model group and EA group.The FD model was replicated by tail clamping,irregular diet,and filling the stomach with ice of Saline Solution.After modeling,the EA group received acupuncture treatment for 1 time day,30 minutes time,for 14 days.Recording the general state of the rat.Detection of locomotion and catatonia in rats by open field test.HE staining to observe the morphology and inflammation of gastric mucosa in rats.PCR detection of mRNA expression of 5-hydroxytryptamine3 receptor and corticotropin-releasing hormone in rat hypothalamus.Detection of corticotropin-releasing hormone receptor-2 and NOD-like receptor protein 6 inflammasome protein expression in rat duodenum by Western blotting.Alcian blue staining was used to detect the expression of rat duodenum goblet cells.Results Compared with the blank group,the general state,distance,speed,duodenum CRHR2 and NLRP6 proteins in the model group were significantly decreased(P<0.05),the hypothalamic 5-HT3R and CRH mRNA were significantly increased(P<0.05).Compared with the model group,the general state,distance,speed,expression of CRHR2,NLRP6 protein and goblet cells in the duodenum of rats in the EA group were significantly increased(P<0.05),the 5-HT3R and CRH mRNA in the hypothalamus were significantly decreased(P<0.05).In the model group,the connective tissue of the gastric mucosa was loosely arranged,the submucosa had mild edema,and there were some lymphocytes.The connective tissue of the gastric mucosa of the rats in the blank group and the electroacupuncture group was closely arranged,and there was no obvious proliferation of interstitial cells and no inflammatory cells.Conclusion EA can increase the expression of CRHR2,NLRP6 protein and goblet cells in the duodenum,and inhibit the expression of 5-HT3R and CRH in the hypothalamus.EA can improve gastrointestinal motility and locomotion,relieve anxiety,repair the mucosal barrier of the defective intestine,and restore the function of the Hypothalamic-Pituitary-Adrenal axis.
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Objective:To investigate the effect of time-restricted feeding (TRF) on bone marrow fat of proximal femur in obese rats induced by high-fat diet (HFD) using proton density fat fraction (PDFF).Methods:Totally 30 male Sprague-Dawley rats were stratified and randomly sampled into 6 subgroups according to body weight, with 5 rats each. Then 2 subgroups were combined into one group, and there were totally 3 groups. The rats in the control group were fed with normal diet, and the rats could eat as much as they wanted for 24 h; the rats in the HFD group were fed with high-fat diet, and the rats could eat as much as they wanted for 24 h; the rats in the HFD+TRF group were fed with high-fat diet only between 9 AM (2 h after light) and 17 PM. One subgroup of rats from each group was examined with MRI on the femur on day 28 of the experiment, and the other subgroup from each group was examined on day 56 to measure the bone marrow PDFF of proximal femur based on mDixon-Quant quantitative sequence images. The rats were executed at the end of the scanning period, and blood samples were collected to measure serum levels of leptin. One-way ANOVA or Kruskal-Wallis H test was used to compare the differences in body weight, PDFF, and serum levels of leptin among 3 groups. The LSD- t test was used for multiple comparisons. Results:On day 28 of the experiment, the differences in body weight, PDFF, and serum leptin among the 3 groups of rats were not statistically significant ( P>0.05). On day 56, the bone marrow PDFF of proximal femur of the rats in the control group, HFD group, and HFD+TRF group were (7.2±1.4)%, (9.7±2.4)%, and (11.2±3.6)%, respectively. The differences in body weight, PDFF, and serum levels of leptin among the 3 groups of rats were statistically significant ( F=6.95, P=0.010, F=5.98, P=0.007, F=4.54, P=0.034). The results of multiple comparisons showed that the body weight in the HFD group was higher than those in the control group (LSD- t=52.96, P=0.036) and the HFD+TRF group (LSD- t=82.74, P=0.003). The values of bone marrow PDFF of proximal femur in the HFD+TRF group was higher than that in the control group (LSD- t=4.01, P=0.012). The serum levels of leptin in the HFD group were higher than those in the control group (LSD- t=1.45, P=0.030) and the HFD+TRF group (LSD- t=1.62, P=0.018). Conclusion:TRF induces an increase in the values of bone marrow PDFF of proximal femur in conjunction with weight loss in obese rats induced by HFD, and the increase in bone marrow fat may be related to the decrease in serum leptin.
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Objective:To explore the feasibility of chemical exchange saturation transfer (CEST) imaging at 3.0 T MRI in quantifying renal redox metabolism in vitro models and experimental animals.Methods:Redox metabolites in vitro models with physiological concentrations were prepared, including reduced metabolites (glutamate, alanine, glutathione) and oxidized metabolites (2-ketoglutarate, pyruvate, glutathione disulfide, ammonium hydroxide). CEST examinations were performed at 3.0 T MRI. The imaging parameters were as follows: CEST images with different saturation pulse intensity (B 1) (1, 2, 3, 4 μT) and a fixed radio frequency (RF) duration of 2 000 ms; CEST images with different RF durations (1 500 and 2 000 ms) were acquired with a fixed B 1 value of 2 μT to obtain the optimal scanning parameters. CEST examinations with optimized parameters were performed on the left kidneys of seven healthy rabbits, and the differences in magnetic resonance ratio asymmetry (MTR asym) between rabbit renal cortex and outer medulla were measured. A paired t-test was used to compare the differences. Results:The optimal B 1 for CEST examination of redox metabolites was 2 μT, and the optimal RF duration was 2 000 ms. The MTR asym peaks of glutathione disulfide, glutathione, glutamic acid, and alanine were at 3.75, 3.5, 3, and 1.5 ppm, respectively. The MTR asym peaks of pyruvate, 2-ketoglutarate, and ammonium hydroxide were at 1 ppm. The MTR asym peak values of reduced metabolites were higher than those of oxidized metabolites. When the B 1 value was 2 μT and the RF duration was 2 000 ms, the MTR asym signal of the renal cortex was (2.60±1.10) %, (2.86±1.32) %, (3.04±1.06) %, and (2.98±0.91) % at 1, 3, 3.5, and 3.75 ppm, respectively. The MTR asym signal of the outer medulla was (1.00±0.56) %, (2.43±0.94) %, (2.29±0.88) % and (1.98±0.58) %, respectively. The MTR asym signal of the renal cortex was higher than that of the outer medulla, and the differences were statistically significant ( t=3.04, P=0.023; t=2.56, P=0.043; t=3.50, P=0.013; t=3.45, P=0.014). Conclusion:CEST imaging at 3.0 T MRI can be used to quantitatively evaluate redox metabolism of healthy rabbit kidneys in vitro model and normal experimental rabbits.
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ObjectiveTo investigate the effect of high-frequency irreversible electroporation （H-FIRE） in the ablation of pig pancreatic tissue. MethodsLaparotomy was conducted in this study， and needle electrodes were used to release electric pulses in 12 pigs. Three sets of parameters were established for ablation at the low， medium， and high values of field strength （1 000 V/cm， 1 500 V/cm， and 2 500 V/cm）. The groups were compared in terms of the data including postoperative recovery， ablation area， and histopathological features to validate the safety and efficacy of H-FIRE in the ablation of porcine pancreatic tissue. The paired t-test was used for comparison of continuous data between two groups. ResultsAll pigs in the experiment survived and showed a good effect of ablation. The histopathological analysis of all groups showed thorough and effective ablation， with a clear boundary between the ablated area and the normal tissue area. The mean ablation area in the low， medium， and high field strength groups was 30.96±3.73 mm2， 51.93±25.26 mm2， and 108.90±55.23 mm2， respectively， and the high and medium field strength groups had a significantly larger ablation area than the low field strength group （both P<0.05）， while there was no significant difference in ablation area between the medium and high field strength groups （P>0.05）. ConclusionH-FIRE ablation is safe and effective for porcine pancreatic tissue under specific ablation parameters.
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Aim: This study was performed to compare two different rat defect models (critical calvaria defects versus guided bone regeneration in the mandibular ramus) used to evaluate bone repair in grafted areas. Methods: A total of 12 rats were allocated in two groups according the experimental model used to evaluate the bone repair in grafted areas: a critical sized-calvaria defect of 5 mm filled with bone graft (n=6) and a mandibular ramus filled with the bone graft associated with a Teflon dome-shaped membrane (n=6). Both groups were grafted with deproteinized bovine bone graft. After 60 days, the animals were euthanized and the samples obtained were submitted to histomorphometry analysis to evaluate the relative amount of bone, remaining bone substitute, and soft tissue within the grafted areas. Results: No differences were observed between the preclinical models evaluated in relation to the amount of bone tissue formation (19.93 ± 4.55% in calvaria vs. 21.00 ± 8.20% in mandible). However, there was a smaller amount of soft tissue (43.20 ± 10.97% vs. 57.79 ± 7.61 %; p<0.01) and a greater amount of bone substitute remaining (35.80 ± 5.52% vs. 22.28 ± 4.36 %; p<0.05) in the grafted areas in the mandible compared to calvaria defect. Conclusion: Preclinical models for the analysis of bone repair in grafted areas in the mandible and critical sized-calvaria defects showed different responses in relation to the amount of soft tissue and bone substitute remnants
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Animals , Rats , Bone Regeneration , Bone Substitutes , Animal Experimentation , HistologyABSTRACT
Abstract Experimental research with the use of animal models to represent a specific reality is a target of criticism by the population. This study analyzed the knowledge of elementary and high school students on experimental/animal research methods, official ethical guidelines/laws, and regulatory institutions. In total, 35 students answered an informative questionnaire with objective questions about the subject. Only 18 students (Group 1), attended a lecture on the subject. The questionnaire was reapplied to all 35 students. The analysis of the first questionnaire round showed rejection by 51.4% of the students toward the methods used in experimental research. Significant changes in answer patterns between the first and second evaluations were observed, with a decrease in the number of students who strongly disagree with the methods used in experimental research in Group 1 (38.8%) compared to Group 2 (88.2%). These data suggest that educational activities could increase the acceptance of experimental research by the community.
Resumen La investigación experimental en animales para representar una realidad específica es criticada por la población. Este estudio analizó el conocimiento de los alumnos de la primaria y la secundaria sobre los métodos de investigación experimental/animal, los lineamientos/leyes éticas oficiales e instituciones que reglamentan. Un cuestionario con preguntas objetivas sobre el tema fue respondido por 35 alumnos. Solo 18 alumnos (Grupo 1) asistieron a una conferencia sobre el tema. El cuestionario se aplicó nuevamente a 35 alumnos. El primer cuestionario mostró que el 51,4% de los alumnos rechazaban los métodos utilizados en la investigación experimental. Hubo cambios significativos en las respuestas entre la primera y la segunda evaluación, con menor número de alumnos que estaban totalmente en desacuerdo con los métodos de la investigación experimental en el Grupo 1 (38,8%) en comparación con el Grupo 2 (88,2%). Por tanto, las actividades educativas pueden incrementar la aceptación comunitaria de la investigación experimental.
Resumo A pesquisa experimental com uso de modelos animais para representar uma realidade específica é alvo de críticas por parte da população. Este estudo analisou o conhecimento de alunos do ensino fundamental e médio sobre métodos de pesquisa experimental/animal, diretrizes/leis éticas oficiais e instituições reguladoras. Ao todo, 35 alunos responderam a um questionário informativo com perguntas objetivas sobre o assunto. Apenas 18 alunos (Grupo 1) assistiram a uma palestra sobre o tema. O questionário foi aplicado novamente aos 35 alunos. A análise do primeiro questionário mostrou rejeição por 51,4% dos estudantes em relação aos métodos utilizados na pesquisa experimental. Observaram-se mudanças significativas nos padrões de resposta entre a primeira e a segunda avaliação, com diminuição do número de alunos que discordam totalmente dos métodos utilizados na pesquisa experimental no Grupo 1 (38,8%) em relação ao Grupo 2 (88,2%). Esses dados sugerem que atividades educativas podem aumentar a aceitação da pesquisa experimental pela comunidade.
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Vaccines , Ethics Committees , Animal Experimentation , Drug DevelopmentABSTRACT
Propósito/Contexto. En el presente trabajo se llevará a cabo una reinterpretación de las tres erres (3R) propuestas por William Russell y Rex Burch (reemplazo, reducción y refinamiento), con el objetivo de ampliar su alcance y mejorar las prácticas de experimentación con animales no humanos. Metodología/Enfoque. Se revisará el sentido que le dieron Russell y Burch a las 3R y se evaluará el modo en que cada una de ellas podría redefinirse o complementarse a la luz de las prácticas científicas, las posibilidades técnicas y los conocimientos bioéticos actuales vinculados al uso de animales en investigación. Resultados/Hallazgos. El artículo mostrará que 1) no solo habrían de reemplazarse animales, sino también las ideas equívocas que tenemos, tanto sobre ellos, como sobre la importancia de la educación bioética en la formación científica, 2) que la reducción, además de referirse al número de sujetos utilizados en cada experimento, debería servir para acabar con investigaciones innecesarias, repetitivas y superfluas, así como con algunos persistentes equívocos sobre el modo de operar de la ciencia y 3) que el refinamiento tendría que salir del espacio experimental para extenderse al modo en que pensamos sobre ética animal en el ámbito de la investigación. Discusión/Conclusiones/Contribuciones. El trabajo da cuenta de la importancia que tiene la incorporación del conocimiento bioético contemporáneo en las prácticas de experimentación con animales para mejorar el carácter reflexivo y ético de la ciencia.
Purpose/Background. In the present work, a reinterpretation of the 3Rs (3Rs) proposed by William Russell and Rex Burch (Replacement, Reduction and Refinement) will be carried out with the aim of broadening its scope and improving nonhuman animal experimentation practices. Methodology/Approach. The meaning given by Russell and Burch to the 3Rs will be reviewed and the way in which each of them could be redefined or complemented in the light of current scientific practices, technical possibilities and bioethical knowledge related to the use of animals in research will be evaluated. Results/Findings. The article will show that 1) not only animals should be replaced, but also the misconceptions we have, both about them and about the importance of bioethics education in scientific training, 2) that the reduction, in addition to the number of subjects used in each experiment, should serve to end unnecessary, repetitive and superfluous research, as well as some persistent misconceptions about the way science operates, and 3) that refinement should go beyond the experimental space to extend to the way we think about animal ethics in the research setting. Discussion/Conclusions/Contributions. The paper reports on the importance of incorporating contemporary bioethical knowledge into animal experimentation practices to enhance the reflexive and ethical character of science.
Objetivo/Contexto. Neste documento, uma reinterpretação dos 3Rs (3Rs) propostos por William Russell e Rex Burch (Substituição, Redução e Refinamento) será realizada com o objetivo de ampliar seu escopo e melhorar as práticas não-humanas de testes em animais. Metodologia/ Abordagem. Revisaremos o significado dado por Russell e Burch aos 3Rs e avaliaremos como cada um deles poderia ser redefinido ou complementado à luz das práticas científicas atuais, possibilidades técnicas e conhecimentos bioéticos relacionados ao uso de animais na pesquisa. Resultados/Descobertas. O artigo mostrará que 1) não somente os animais devem ser substituídos, mas também conceitos errôneos sobre eles e a importância da educação bioética no treinamento científico, 2) que a redução, além do número de sujeitos utilizados em cada experimento, deve servir para eliminar pesquisas desnecessárias, repetitivas e supérfluas, assim como alguns conceitos errôneos persistentes sobre a maneira como a ciência funciona, e 3) que o refinamento deve se estender além do espaço experimental para a maneira como pensamos sobre a ética animal na pesquisa. Discussão/Conclusões/Contribuições. O artigo explica a importância de incorporar o conhecimento bioético contemporâneo nas práticas de experimentação animal para realçar o caráter reflexivo e ético da ciência.
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Las Ciencias Médicas y Biológicas requieren, prioritariamente, que la investigación y la experimentación sean desarrolladas sobre organismos completos (los modelos animales). Su utilización ha permitido desarrollar innumerables ensayos preclínicos para evaluar los mecanismos patógenos y terapéuticos de diversas enfermedades, así como el estudio de las causas, naturaleza y cura de múltiples desórdenes de la salud humana. En este trabajo se muestra una panorámica general de los biomodelos de hipertensión arterial donde se describen: conceptos, características, origen, importancia, utilidad y procedimientos experimentales durante su fase de inducción. También se pondera la justificación de los biomodelos empleados en los estudios preclínicos de esta enfermedad. De igual forma, se describen los antecedentes para medir las alteraciones, las técnicas y los métodos directos e indirectos de medición de la presión arterial, la cual fue provocada experimentalmente en los animales de laboratorio para realizar los estudios de hipertensión humana.
Medical and biological sciences require, as a priority, that research and experimentation be carried out on complete organisms (animal models). Its use has allowed the development of innumerable preclinical tests to evaluate pathogenic and therapeutic mechanisms of various diseases, as well as to study the causes, nature and cure of multiple human health disorders. In this work, we show a general overview of arterial hypertension biomodels where concepts, characteristics, origin, importance, utility and experimental procedures during their induction phase are described. The justification of the biomodels used in preclinical studies of this disease is also considered. Antecedents are also described to measure alterations, techniques and direct and indirect methods of measurement of arterial pressure, which was provoked experimentally in the laboratory animals to carry out the studies of human hypertension.
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Rats , Models, Animal , Animal Experimentation , Hypertension , Animals, LaboratoryABSTRACT
Objective:To explore the effect of fecal microbiota transplantation (FMT) on the formation of renal calcium oxalate crystals in SD rats induced by oxalate mixed diet.Methods:Six male guinea pigs were fed with standard guinea pig chow for 1 month and then given a 5% oxalate diet for 14 d. The guinea pigs on the standard chow were labeled as the standard chow guinea pig (GSC group) and those on the high oxalate diet for 14 d were labeled as the guinea pig group on the high oxalate diet (GOD group). The feces of guinea pigs in the GSC and GOD groups were collected using metabolic cages. Twenty-four male SD rats were randomly divided into standard chow (SC) group, oxalate diet(OD)+ phosphate buffered saline gavage group (OD+ PBS group), OD+ FMT group and SC+ FMT group. Among them, the SC group and SC+ FMT group were fed with standard chow. The OD+ PBS group and OD+ FMT group were fed with 5% oxalate content chow. The OD+ FMT and SC+ FMT groups were given GOD group guinea pig fecal filtrate gavage for 7 days. The 24 h urine and feces of rats in each group were collected, and the intestinal microbiota of rats and guinea pigs were detected by 16sRNA detection. The urinary oxalate excretion was detected by high performance liquid chromatography. The rats and kidneys were weighed and the renal index was calculated. HE staining was used to observe the histological morphological changes of rat kidney tissue, the calcium oxalate crystal deposition in renal tissues was detected by Pizzolato staining.Results:The relative abundance of bacteria from a total of 11 families, including Muribaculaceae family and Bifidobacteriaceae family, was significantly increased in the intestinal tract of guinea pigs (GOD) from the high oxalate diet group compared to guinea pigs (GSC) from the standard chow group. The microbial diversity of the intestinal microbiota of the rats in the OD+ PBS group was reduced compared to the SC group, and the microbial diversity of the intestinal microbiota of the rats in the OD+ FMT group was restored compared to the OD+ PBS group. When given a standard chow, the intestinal microbiota of rats receiving FMT deviated from that of normal rats and was more similar to that of guinea pigs fed a high oxalate diet. In the OD+ FMT group, bacteria from a total of 18 families, including Muribaculaceae family, Erysipelotrichaceae family and Bifidobacteriaceae family, were significantly enriched, and FMT activated the intestinal microbial network represented by bacteria from Muribaculaceae family. The renal index of rats in the OD+ PBS group was significantly increased compared to the SC group (7.63±0.67 vs. 6.12±0.53, P<0.05), whereas the renal index of rats in the OD+ FMT group was significantly decreased in comparison to the OD+ PBS group (6.53±0.64 vs. 7.63±0.67, P<0.05). Urinary oxalate excretion of rats in the SC group, the OD+ PBS group, and the OD+ FMT group were (0.61±0.05), (0.89±0.04) and (0.72±0.04) μmol/ml, respectively. In the rats of the SC group no calcium oxalate crystals were seen in the kidney (0 score) and more calcium oxalate crystals were detected in the OD+ PBS group (4.83±0.41 score). The OD+ FMT group showed significantly lower calcium oxalate crystallization scores (3.17 ± 0.75 score, P<0.01) compared to the OD+ PBS group. Conclusions:FMT activated the microbial network represented by bacteria from the family Muribaculaceae in the rat intestine, significantly reduced urinary oxalate excretion and renal calcium oxalate crystal deposition in rats on a high oxalate diet.
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Objective:To explore MRI T 2-mapping and blood oxygenation level dependent (BOLD) to evaluate the functional changes of paraspinal muscle in rats with discogenic low back pain (DLBP) after swimming. Methods:Totally 54 female 1-month-old SD rats were selected, which were divided into 3 groups by random number table method, sham operation (Sham) group, DLBP non-swimming group and DLBP swimming group, with 18 rats in each group. Under the guidance of X-ray fluoroscopy, the L4/5 and L5/6 intervertebral discs of the rats in the DLBP non-swimming group and DLBP swimming group were punctured by the posterior approach, and establishment of DLBP rat model by destroying nucleus pulposus, and only paraspinal muscles at the same level were punctured in the Sham group. After modeling, the DLBP swimming group received swimming exercise intervention for 5 consecutive days (30 min/d), while the DLBP non-swimming group and Sham group did not receive any rehabilitation exercise intervention. Each group was divided into 3 time point subgroups on average, the T 2-mapping and BOLD sequences were scanned at 30, 90 and 180 days after modeling to obtain the T 2 value, R 2* value of the paraspinal muscles, and the paraspinal muscles at the modeling level were taken for immunofluorescence staining, and the fluorescence intensity of myosin heavy chain (MYH)1 (type Ⅱ muscle fiber) and MYH7 (type I muscle fiber) was analyzed. One-way analysis of variance was used for comparison among the 3 groups, and the Bonferroni method was used for multiple comparisons, and Pearson correlation coefficient was used to evaluate the correlation between quantitative MRI parameters T 2 value, R 2* value and MYH1, MYH7 immunofluorescence intensity of rat paraspinal muscles at 180 days after modeling. Results:At 30 days after modeling, there was no significant difference in T 2 value and R 2* value among the 3 groups (all P>0.05). At 90 days after modeling, the T 2 value of the DLBP swimming group was higher than that of the DLBP non-swimming group, and the T 2 value of the DLBP non-swimming group was lower than that of the Sham group (all P<0.05), and there was no significant difference in the R 2* value among the 3 groups ( P>0.05). At 180 days after modeling, the T 2 value of the DLBP swimming group was higher than that of the DLBP non-swimming group, and the R 2* value was lower than that of the DLBP non-swimming group; the T 2 value of the DLBP non-swimming group was lower than that of the Sham group, and the R 2* value was higher than that of the Sham group (all P<0.05). At 30 and 90 days after modeling, there was no significant difference in the expressions of MYH1 and MYH7 among the 3 groups (all P>0.05). At 180 days after modeling, the expression of MYH1 decreased and the expression of MYH7 increased in the DLBP swimming group compared with the DLBP non-swimming group; the expression of MYH1 increased and the expression of MYH7 decreased in the DLBP non-swimming group compared with the Sham group (all P<0.05). At 180 days after modeling, the T 2 value had a moderate negative correlation with the fluorescence intensity of MYH1 ( r=-0.511, P=0.043), and a moderate positive correlation with the fluorescence intensity of MYH7 ( r=0.564, P=0.023); R 2* value was moderate positive correlated with the fluorescence intensity of MYH1 ( r=0.625, P=0.010), and moderate negative correlated with the fluorescence intensity of MYH7 ( r=-0.653, P=0.006). Conclusions:Swimming exercise can improve the reduction of water content and perfusion in the paraspinal muscles of DLBP rats, and reduce the transformation of muscle fibers from type Ⅰ to type Ⅱ, the changes of T 2 and R 2* value can reflect the transformation of paraspinal muscle fiber types to a certain extent.
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Objective:To develop an improved wireless intelligent capsule cystoscope (WCE)for dynamic detection of bladder mucosa in a pig model.Methods:The WCE was introduced into a healthy experimental pig that under general anesthesia via urethra by applying an improved device. Multi-angle images of the bladder mucosa were then obtained by controlling the position of capsule cystoscope with an external magnetic field system. The shutter speed of the WCE was 2.5 fps and was automatically converted to 1.5 fps 30 minutes after initiation. The Vue software was utilized to download the shoot pictures which were former received by a computer via wireless transmission. The pig was roused and sent to the pigpen, without limitations in moving. The improved WCE was connected with a 2 cm thread. 12 hours later, the dilated sheath was inserted again, and the capsule was removed by a foreign body forceps under observation of a ureteroscopy.Results:The WCE was successfully placed and removed from the pig's bladder with the application of the improved devices. Over 20 thousand images that with 60K pixels of bladder mucosa were captured by the WCE at various angles within 12 hours, which revealed the process of urine filling and excreting in a time-dependent way. No notable adverse effects (bleeding, urinary tract injury, etc) were noted during the process of cystoscope placement, image acquisition, transmission, and removal.Conclusion:This study developed a novel WCE that could dynamically, intelligently and accurately monitor all aspects of the pig bladder mucosa, and has preferable application prospect.
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Objective:To explore the feasibility of anastomosis between bladder and intestine of experimental rabbit by drag anastomosis.Methods:In this study 40 Japanese big-eared rabbits were randomly divided into two groups through random number table, the experimental group and the control group, each group with 20 rabbits. In the experimental group, the bladder neck was fixed to the catheter and then the catheter was drawn outward. With the traction of the catheter, the bladder neck was anastomosed with the distal intestinal tube by means of suture free. The control group was anastomosed by regular interrupted suture of bladder and intestine. The operation time, anastomosis time, intraoperative blood loss, postoperative urinary leakage rate and postoperative anastomotic healing of rabbits in the two groups were compared.Results:The operation time of the experimental group was shorter than that of the traditional interrupted suture anastomosis group [(33.26±2.79)min vs. (35.25±1.83)min, P=0.014]. The anastomosis time of the experimental group was significantly shorter than that of the traditional interrupted suture anastomosis group[(7.55±1.2)min vs. (8.65±1.03 min), P=0.005]. The intraoperative blood loss in the experimental group was similar to that in the control group[(6.47±2.41) ml vs. (6.75±1.83) ml, P=0.691]. The event of contrast media extravasation occurred in 2 of the 10 experimental rabbits after receiving cystography in the experimental group, and the urinary leakage rate was 20%(2/10). In the control group, contrast media extravasation occurred in 1 of the 9 experimental rabbits after receiving cystography, and the urinary leakage rate was 11.1%(1/9), and the difference of the two groups was not statistically significant ( P=0.348). Anastomotic healing score was (2.0±0.7) in the experimental group, and (2.1±0.74) in the control group ( P=0.767). Conclusions:The bladder-intestine drag-and-bond anastomosis technique, with significantly shorter anastomosis time, was feasible, easy and convenient. Our research provides an experimental and theoretical basis for the clinical application of drag-and-bond anastomosis technique in clinic.
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Objective:To construct a C57BL/6 mouse model of simulating transurethral thulium laser vaporization prostatectomy.Methods:Twelve male C57BL/6 mice were selected to undergo transvesical vaporization resection of the urothelium covering the urethra of the prostate using thulium laser. The urethral tissue of the prostate was retrieved on the 1st, 3rd, 5th, and 7th days after the surgery. HE staining was used to observe the process of re-epithelialization of the urethral wound of the prostate. Immunohistochemical (IHC) staining was used to detect whether the re-epithelialized cells of the urethral wound of the prostate expressed urothelin Ⅲ (UPⅢ).Results:On the first day after surgery, HE staining showed complete destruction to the urothelium covering the urethra of the prostate, with a large amount of coagulative necrotic tissue on the wound surface, and IHC staining showed no expression of UPⅢ on the wound surface. On the 3rd day after surgery, HE staining showed that there were still no regenerated epithelial cells on the wound surface, with coagulation necrosis tissue significantly reduced, and the urethral cavity was clearly visible. And IHC staining showed no expression of UPⅢ on the wound surface. On the 5th day after surgery, HE staining showed 1-2 layers of regenerated epithelial cells lacking cell polarity on the wound surface, and IHC staining showed that the regenerated epithelial cells expressed UPⅢ. On the 7th day after surgery, HE staining showed 4-6 layers of polar regenerated epithelial cells on the wound surface, and IHC staining showed the multiple layers of regenerated epithelial cells expressing UPⅢ.Conclusions:Based on the simulation of transurethral thulium laser vaporization resection of the prostate, the thulium laser and ultra micro endoscope system were used to vaporize the urothelium covering the urethra of the prostate, and the process of urethral re-epithelialization of the prostate can be observed after surgery. The establishment of the C57BL/6 mouse model simulating thulium laser vaporization prostatectomy provides a new research platform for studying the mechanism of wound repair after prostatectomy.
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In recent years, collagen peptides (CP) have become a research hotspot in delaying chronological skin aging. Animal experiments have shown that CP can repair chronologically aged animal skin by promoting collagen synthesis, inhibiting collagen degradation, and increasing antioxidant enzyme activity. Cell experiments showed that CP can promote proliferation of fibroblasts and synthesis of collagen and elastin by stimulating nuclear factor-κB signaling pathway and transforming growth factor-β/drosophila mothers against decapentaplegic signaling pathway. Clinical studies have demonstrated that long-term oral supplement with CP or CP in combination with other antioxidant active substances can increase the skin moisture content and reduce transepidermal water loss, improve skin wrinkles and elasticity, as well as improve the skin collagen fiber structure, dermal and epidermal quality and the overall condition of facial skin. This review summarizes recent studies on mechanisms underlying chronological skin aging and mechanisms of action of CP in repairing chronologically aged skin, in order to provide a theoretical basis for further clinical research into and application of CP in repairing chronologically aged skin.
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Objective:To deeply explore the potential mechanism of Kangmin Zhisou Granules in the treatment of bronchial asthma through network pharmacology method; To verify it with animal experiments.Methods:The active components and corresponding target information of Kangmin Zhisou Granules were screened with the help of BATMAN-TCM database, and the related disease targets of bronchial asthma were obtained through GeneCards and OMIM databases. The drug targets and bronchial asthma targets were intersected and imported String database was used to establish PPI network. Cytoscape 3.9.1 software was used to draw the network diagram of "Chinese materia medica-active components-intersection targets" and the core targets were screened. GO and KEGG enrichment analysis was performed on the core targets using DAVID database. A mouse model of asthma induced by ovalbumin was prepared. After the intervention of Kangmin Zhisou Granules, the pathological changes of mouse lung tissue were observed, and the contents of serum TNF-α, IL-6, IL-1 β were detected by ELISA.Results:Totally 240 active components and 1 364 potential targets were obtained from Kangmin Zhisou Granules. Tumor necrosis factor (TNF), interleukin-6 (IL-6), protein kinase B (AKT1), albumin (ALB), interleukin 1-beta (IL-1β) and other 11 core targets were obtained after screening. The results of GO enrichment analysis showed that the treatment of bronchial asthma by Kangmin Zhisou Granules mainly involved the positive regulation of protein phosphorylation, the regulation of inflammatory response, lipopolysaccharide response and other biological processes, as well as TNF, activated protein kinase (MAPK), interleukin-17 (IL-17) and other signaling pathways. Animal experiments confirmed that Kangmin Zhisou Granules could reduce the expression levels of TNF-α, IL-6 and IL-1β in serum ( P<0.05), and reduce the infiltration of inflammatory cells in the lung tissue of mice, thereby relieving asthma symptoms. Conclusion:Kangmin Zhisou Granules may exert anti-inflammatory effects by acting on TNF-α, IL-6, IL-1β and other targets to alleviate asthma symptoms.
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Abordagens interdisciplinares vêm ganhando maior reconhecimento e destaque nas comunidades de saúde humana e animal, principalmente pela (re)emergência de diversas doenças infecciosas que emanam da interface humano-animal-ambiente. A raiva, zoonose grave, considerada endêmica no Brasil e globalmente negligenciada, é um exemplo. Tanto a vigilância epidemiológica quanto a confirmação dessa doença dependem do diagnóstico laboratorial, que é realizado, frequentemente, por meio dos testes de Imunofluorescência Direta (IFD) e de Isolamento Viral em Camundongo (IVC), via inoculação intracerebral da amostra suspeita em camundongos lactentes ou desmamados. Entretanto, recentemente, a Organização Mundial da Saúde reconheceu a Transcrição Reversa seguida da Reação em Cadeia da Polimerase (RT-PCR) como uma técnica primária válida para esse diagnóstico, podendo ser empregada como alternativa ao uso de animais, evitando sofrimento e morte. Esta dissertação apresenta uma discussão sobre as implicações técnicas e éticas da (não) adoção desse método substitutivo, considerando que todos os animais devem ser respeitados e entendidos como sujeitos singulares em suas percepções do mundo, não como objetos de pesquisa. Esse fato corrobora a necessidade de novas perspectivas que ressignifiquem nossas relações com os animais não humanos, o que é primordial para o estabelecimento de mudanças sistêmicas, de caráter ético-político, que visem o fim da instrumentalização animal e de seu uso no âmbito científico, bem como de qualquer forma de opressão.
Interdisciplinary approaches have been gaining greater recognition and prominence in the human and animal health communities, mainly due to the (re)emergence of several infectious diseases that emanate from the human-animal-environment interface. Rabies is an example, considered a serious zoonosis endemic in Brazil and globally neglected. Both epidemiological surveillance and confirmation of this disease depend on laboratory diagnosis, which is usually performed by the direct fluorescent antibody test (DFAT) and the mouse inoculation test (MIT) via intracranial inoculation of the suspected sample into suckling or weanling mice. However, the World Health Organization recently recognized the reverse transcription polymerase chain reaction (RT-PCR) as a valid primary technique for this diagnosis, which can replace the use of animals, avoiding suffering and death. This study presents a discussion about the technical and ethical implications of (not) adopting this alternative method, considering that all animals must be respected and understood as unique beings with their perceptions of the world, not as objects of research. It also further corroborates the need for new perspectives that reframe our relationships with non-human animals, which is fundamental for the implementation of systemic ethical-political changes, aiming at the end of animal instrumentalization and use in scientific research, as well as all forms of oppression.
Subject(s)
Humans , Animals , Rabies , Bioethics , Animal Experimentation , Ethics, Research , Brazil , Animal Use AlternativesABSTRACT
Objective:To explore the value of proton density fat fraction(PDFF) based on histogram analysis for quantification hepatic steatosis and fibrosis in rabbit model and the interference of hepatic fibrosis to the evaluation of hepatic steatosis with PDFF.Methods:From March to November 2020, 135 New Zealand white rabbits were randomly divided into control group ( n=30) and experimental group ( n=105) using a random number table. The volume ratio of CCl 4 and olive oil was 1∶1 to prepare 50% CCl 4 oil solution, and experimental rabbits were subcutaneously injected with the oil solution. An equal dose of normal saline was subcutaneously injected for control group rabbits. At the end of the 4 th, 8 th, and 12 th week, 35 in the experimental group and 10 rabbits in the control group were randomly selected to conduct the mDixon-Quant scanning, and histogram analysis of PDFF was analyzed including volume, mean, median, standard deviation, 25 th, 50 th, 75 th, 90 th quantile, skewness, kurtosis, entropy and inhomogeneity. After the examination, the rabbits were sacrificed and the liver percentage of steatosis (PSH) and fibrosis (POF) were recorded by semi-quantitative analysis. Spearman correlation analysis was used to correlate PDFF with PSH and POF. Multiple linear regression analysis was used to determine independent PDFF histogram parameters for evaluating PSH and POF. A receiver operator characteristic (ROC) curve was used to assess the diagnostic accuracy of PDFF for discriminating mild from moderate-severe hepatic steatosis and mild from moderate-severe hepatic fibrosis with median of PSH or POF for dichotomy, and DeLong test was used to compare the area under the curve (AUC). With the correction of hepatic fibrosis, correlation coefficient and AUC were compared of PDFF for discrimination mild from moderate-severe hepatic steatosis. Results:The PDFF mean, median, standard deviation, 75 th, 90 th showed correlation with PSH ( r=0.558, 0.522, 0.319, 0.723, 0.646, -0.589, all P<0.05). The entropy and 75 th were independent parameters for evaluating PSH (β=2.347, -5.960, P=0.018, 0.001). The PDFF 75 th was the optimal parameter for discriminating mild from moderate-severe hepatic steatosis with AUC=0.915 ( P=0.001). The PDFF volume, mean, median, standard deviation, 75 th, 90 th, entropy showed correlation with POF ( r=0.355, 0.393, 0.376, 0.298, 0.485, 0.426, -0.681, all P<0.05). The entropy, standard deviation and volume (β=-11.041, 1.356, 0.190, P=0.001, 0.026, 0.016) were independent parameters for evaluation of hepatic fibrosis, and the entropy was the optimal parameter for hepatic fibrosis (AUC=0.771, P=0.001). The correlation between PSH and PDFF 75 th was less pronounced when fibrosis was present ( r=0.512, P=0.001) than when fibrosis was absent ( r=0.751, P=0.002). The PDFF 75 th showed a significant difference in discriminating mild hepatic steatosis from moderate-severe hepatic steatosis after correction of POF (AUC=0.895, 0.950, Z=2.970, P=0.025). Conclusions:PDFF based on histogram analysis provided a noninvasive, accurate estimation of quantification for hepatic steatosis and fibrosis. Hepatic fibrosis reduced the correlation between hepatic steatosis and PDFF and the presence of hepatic fibrosis can confound the quantification of hepatic steatosis with PDFF.
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Objective:To observe the expression of miRNA in retinal tissue of mice with oxygen-induced retinopathy (OIR), and screen miRNAs related to p21 and retinal neovascularization (RNV) formation.Methods:A experimental study. Forty healthy 7-day-old C57BL/6J mice were randomly divided into normal group and OIR group, with 20 mice in each group. The oxygen induced RNV model was constructed in the OIR group, and no treatment was performed in the normal group. At the age of 17 days, the mice were killed and the RNV of mice was observed by retinal fluorescence; the nuclei of vascular endothelium that broke through the inner limiting membrane of retina were counted under light microscope. The retinal tissues were taken for miRNA chip analysis to detect the differentially expressed miRNAs between the normal group and the OIR group. The resulting differential miRNA target genes were subjected to enrichment analysis based on gene annotation (GO) and Kyoto Encyclopedia of genes and genomes (KEGG); miRNAs and pathways that may be related to p21 were screened through Targetscan, MiRanda and MicroT-CDs database alignment. Independent sample t-test was used for pairwise comparison between groups. Results:Compared with the normal group, the area of nonperfusion area, RNV and the number of vascular endothelial nuclei that broke through the inner limiting membrane of the retina in the OIR group increased significantly, differences were statistically significant ( t=18.800, 9.025; P<0.05). Compared with the normal group, there were 54 miRNAs that were statistically differentially expressed in the OIR group, of which 47 were up-regulated and 7 were down-regulated. A total of 13 miRNAs related to p21 were screened from the alignment results of the three databases with the obtained differential miRNAs. According to the difference multiples, they were miR-7218-5p, miR-322-5p, miR-224-5p, miR-335-5p, miR-329-3p, miR-362-3p, miR-532-5p, miR-20b-5p, miR-20a-5p, miR-195a-5p, miR-423-5p, miR-497a-5p, and miR-129-5p. Differential miRNA target gene enrichment analysis yielded 1 112 go entries and 50 KEGG pathways, of which 50 go entries and 13 KEGG pathways were related to p21. Conclusion:13 miRNAs related to p21 were screened out in the OIR model.
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Objective:To observe the expression of miRNA in retinal tissue of mice with oxygen-induced retinopathy (OIR), and screen miRNAs related to p21 and retinal neovascularization (RNV) formation.Methods:A experimental study. Forty healthy 7-day-old C57BL/6J mice were randomly divided into normal group and OIR group, with 20 mice in each group. The oxygen induced RNV model was constructed in the OIR group, and no treatment was performed in the normal group. At the age of 17 days, the mice were killed and the RNV of mice was observed by retinal fluorescence; the nuclei of vascular endothelium that broke through the inner limiting membrane of retina were counted under light microscope. The retinal tissues were taken for miRNA chip analysis to detect the differentially expressed miRNAs between the normal group and the OIR group. The resulting differential miRNA target genes were subjected to enrichment analysis based on gene annotation (GO) and Kyoto Encyclopedia of genes and genomes (KEGG); miRNAs and pathways that may be related to p21 were screened through Targetscan, MiRanda and MicroT-CDs database alignment. Independent sample t-test was used for pairwise comparison between groups. Results:Compared with the normal group, the area of nonperfusion area, RNV and the number of vascular endothelial nuclei that broke through the inner limiting membrane of the retina in the OIR group increased significantly, differences were statistically significant ( t=18.800, 9.025; P<0.05). Compared with the normal group, there were 54 miRNAs that were statistically differentially expressed in the OIR group, of which 47 were up-regulated and 7 were down-regulated. A total of 13 miRNAs related to p21 were screened from the alignment results of the three databases with the obtained differential miRNAs. According to the difference multiples, they were miR-7218-5p, miR-322-5p, miR-224-5p, miR-335-5p, miR-329-3p, miR-362-3p, miR-532-5p, miR-20b-5p, miR-20a-5p, miR-195a-5p, miR-423-5p, miR-497a-5p, and miR-129-5p. Differential miRNA target gene enrichment analysis yielded 1 112 go entries and 50 KEGG pathways, of which 50 go entries and 13 KEGG pathways were related to p21. Conclusion:13 miRNAs related to p21 were screened out in the OIR model.