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1.
Article in Chinese | WPRIM | ID: wpr-880777

ABSTRACT

OBJECTIVE@#To explore the mechanism of @*METHODS@#Healthy male DBA/1 mice were used for CIA modeling. Twenty-five CIA mice with successful modeling and similar arthritis index (AI) scores were randomized equally into model group (CIA), methotrexate (MTX) group, and low-, medium-, and high-dose XWGD groups (0.975, 1.95, and 3.9 g/mL, respectively), with another 5 normal mice as the normal control group. The mice in normal control and CIA groups were given saline once a day, those in MTX group were given 0.1 mg/mL MTX once a week, and those in XWGD groups were treated daily via garage of XWGD containing crude drugs of different doses for 28 consecutive days. The AI score and HE staining were used to evaluate the changes in the joints of the CIA mice. The effect of XWGD on Th1, Th17, MDSC, G-MDSC and M-MDSC cells were evaluated with flow cytometry.@*RESULTS@#Treatment with MTX and different doses of XWGD significantly decreased the AI score of the mice and relieved joint inflammation as compared with the model group (@*CONCLUSIONS@#XWGD can improve joint inflammation in CIA mice by increasing the percentages of G-MDSC cells and decreasing the percentages of M-MDSC, Th1 and Th17 cells, and a high dose of XWGD can produce an equivalent therapeutic effect to methotrexate but with better safety.


Subject(s)
Animals , Arthritis, Experimental/drug therapy , Arthritis, Rheumatoid/drug therapy , Male , Methotrexate , Mice , Mice, Inbred DBA , Th17 Cells
2.
Biol. Res ; 52: 9, 2019. tab, graf
Article in English | LILACS | ID: biblio-1011411

ABSTRACT

BACKGROUND: Fava beans (FBs) have long been used as food, and their principal disadvantage is derived from their haemotoxicity. We hypothesized that FB ingestion alters the intestinal gene expression pattern, thereby inducing an immune response. RESULTS: In-depth sequence analysis identified 769 differentially expressed genes (DEGs) associated with the intestine in FB-treated DBA/1 mouse intestines. The identified genes were shown to be associated with biological processes (such as response to stimulus and immune system processes), human disease pathways (such as infectious diseases, endocrine and metabolic diseases, and immune diseases), and organismal system pathways (such as the digestive system, endocrine system, environmental adaptation, and immune system). Moreover, plasma total immunoglobulin E (IgE), histamine, interleukin (IL)-4 and IL-13 levels were significantly increased when the mice were treated with FBs. CONCLUSIONS: These results demonstrated that FBs affect the intestinal immune response and IgE and cytokine secretion in DBA/1 mice.


Subject(s)
Animals , Male , Mice , Vicia faba/adverse effects , Immunity, Humoral/immunology , Intestinal Mucosa/immunology , Signal Transduction , Reverse Transcriptase Polymerase Chain Reaction , Gene Expression Profiling , Vicia faba/immunology , Favism/etiology , Mice, Inbred DBA
3.
Article in Chinese | WPRIM | ID: wpr-773506

ABSTRACT

OBJECTIVE@#To observe the therapeutic effect of tetramethylpyrazine on immune-mediated bone marrow failure (BMF) induced by different doses of X-ray exposure in C57 mice.@*METHODS@#C57BL6 mice were randomized into 4 groups, including a blank control group and 3 X-ray exposure groups with X-ray exposure at low (5.0 Gy), moderate (5.75 Gy), and high (6.5 Gy) doses. After total body irradiation with 0.98 Gy/min X-ray. The mice as recipient received injections of 4×10 lymphocytes from DBA/2 mice via the tail vein within 4 h. The survival rate of the recipient mice, peripheral blood cell counts, bone marrow nucleated cell count, and bone marrow pathology were examined at 14 days after the exposure. In the subsequent experiment, C57 mice were exposed to 5.0 Gy X-ray and treated with intraperitoneal injection of tetramethylpyrazine at the low (5 mg/mL), moderate (10 mg/mL), or high (20 mg/mL) doses (12 mice in each group) for 14 consecutive days, and the changes in BMF were observed.@*RESULTS@#X-ray exposure, especially at the high dose, resulted in significantly lowered survival rate in the mouse models of BMF at 14 days. As the X-ray dose increased, the mice showed significantly reduced peripheral blood counts of red blood cells, white blood cells, platelets and lowered bone marrow nucleated cell counts with obvious bone marrow congestion and reduction of nucleated cells ( < 0.05 or 0.001). In the mice exposed to 5.0 Gy X-ray, tetramethylpyrazine at the high dose most obviously increased bone marrow nucleated cells ( < 0.01) and red blood cells ( < 0.001), and even at the low dose, tetramethylpyrazine significantly increased the counts of white blood cells ( < 0.05) and platelets ( < 0.01) following the exposure. Tetramethylpyrazine dose-dependently alleviated bone marrow hyperemia, increased bone marrow nucleated cell counts, and lowered Fas protein expression in the bone marrow.@*CONCLUSIONS@#X-ray irradiation at 5.0 Gy is suitable for establish mouse models of immune-mediated BMF. Tetramethylpyrazine promotes bone marrow repair by regulating Fas cell apoptosis signals, which further expands the traditional Chinese medicine theory of "removing blood stasis to create new."


Subject(s)
Animals , Bone Marrow , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Pyrazines , Whole-Body Irradiation
4.
An. acad. bras. ciênc ; 89(1,supl): 635-647, May. 2017. tab, graf
Article in English | LILACS | ID: biblio-886672

ABSTRACT

ABSTRACT The development of DBA/2J mouse strain embryos is nearly 12 h - or 6 somite pairs - delayed as compared to the outbred NMRI mouse embryos of the same age on gestation days (GD) 8-12. To evaluate inter-strain differences in susceptibility to teratogens, dams were treated with methylnitrosourea (MNU, 5 mg/kg body weight i.p.) on defined gestation days (NMRI: GD 9, 91/2 or 10; DBA/2J: GD 10 or 101/2). Skeletal anomalies produced by MNU on both mouse strains varied with the GD of treatment. The pattern of anomalies produced by MNU on a given GD markedly differed between the two mouse strains, yet they were similar -with a few exceptions- when exposures at equivalent embryonic stages are compared. Findings from this study indicated that strain-dependent differences in the developmental stage of mouse embryos of the same gestational age occur, a possibility that has been often neglected when inter-strain differences in susceptibility to developmental toxicants are interpreted.


Subject(s)
Animals , Female , Pregnancy , Rats , Skeleton/abnormalities , Teratogens/toxicity , Somites/abnormalities , Embryonic Development/drug effects , Embryo, Mammalian/abnormalities , Methylnitrosourea/toxicity , Skeleton/drug effects , Skeleton/embryology , Somites/drug effects , Somites/embryology , Embryo, Mammalian/drug effects , Mice, Inbred DBA
5.
Rio de Janeiro; s.n; 2016. 77 p. graf, ilus, tab.
Thesis in Portuguese | LILACS | ID: biblio-983642

ABSTRACT

Fatores que alteram os níveis plasmáticos de substâncias químicas e, por conseguinte, modificam a sua cinética, como por exemplo, a gravidez, podem ter impactos sobre a segurança e eficácia de medicamentos. Em estudo recente, realizado por Carmo (2015), no Laboratório de Toxicologia Ambiental do Departamento de Biologia da Escola Nacional de Saúde Pública da Fundação Oswaldo Cruz (ENSP/FIOCRUZ), foi observado que a concentração plasmática do antimalárico difosfato de primaquina em camundongos fêmeas grávidas DBA/2 era menor do que a concentração do fármaco registrada em igual intervalo de tempo pós-adminstração em camundongos fêmeas não grávidas. Vários estudos sugerem que a diminuição da concentração plasmática de fármacos na gestante pode se dever a um retardo no esvaziamento gástrico e/ou um aumento no volume de distribuição. Alterações do trânsito no trato gastrintestinal podem influenciar diretamente a absorção de fármacos, resultando em absorção mais rápida ou mais lenta. O fármaco analgésico e antipirético paracetamol é absorvido quase que exclusivamente no intestino. Assim a velocidade da sua absorção depende do tempo de esvaziamento gástrico. Fatores tais como alimentação, idade, gravidez e/ou o uso de fármacos que promovem aceleração (metoclopramida) ou o retardo (morfina) da motilidade gastrointestinal, podem influenciar em sua absorção. O objetivo desse trabalho foi desenvolver e padronizar uma metodologia de análise do paracetamol que permitisse investigar o efeito da gravidez sobre o esvaziamento gástrico sobre a cinética de fármacos administrados em pequenos roedores. O método empregado para determinar as concentrações plasmáticas de paracetamol foi a cromatografia em fase líquida de alta eficiência com detector por arranjo de diodos e visualização no ultravioleta (CLAE-DAD-UV), em equipamento Shimadzu Class-VP...


Factors that affect plasma levels of chemicals, and consequently their kinetics, such as pregnancy, can impact on the safety and efficacy of medicines. In a recent study, conducted by Carmo (2015) at the laboratory of Environmental Toxicology (Department of Biological Sciences, National School Public Health, Oswaldo Cruz Foundation -ENSP / FIOCRUZ), it was shown that plasma concentrations of the anti-malarial drug primaquine diphosphate in pregnant female DBA/2 mice were lower than levels found in non pregnant female mice. During pregnancy a delayed gastric emptying and/or an increased volume of distribution may result in lower drug plasma concentrations. Pregnancy-produced changes in the gastrointestinal transit may influence drug absorption. Depending on whether gastric emptying is accelerated or slowed and on the place where drug absorption takes place (stomach or intestines) absorption can be accelerated or slowed. Paracetamol, an analgesic and antipyretic drug, is absorbed almost exclusively in the intestines and is used to investigate the effects of treatment on the gastric emptying rate. Factors such as diet, age, pregnancy or the administration of drugs which accelerate (metoclopramide) or delay (morphine) gastric emptying influence the absorption of paracetamol. The aim of this study was to develop and standardize a methodology to investigate the effect of gastric emptying on the kinetics of drugs administered in small rodents. The methodology used in the analysis of plasma concentrations of paracetamol was High Performance Liquid Chromatography coupled to diode-array detector and visualization on ultraviolet range (HPLC-DAD-UV), using a Shimadzu Class-VP equipment...


Subject(s)
Animals , Acetaminophen , Gastric Emptying , Pregnancy , Rats, Wistar , Chromatography, High Pressure Liquid , Mice , Mice, Inbred DBA , Pharmacokinetics
6.
Article in English | WPRIM | ID: wpr-32082

ABSTRACT

The tongue has 4 kinds of papillae, which are filiform, fungiform (FU), foliate (FO) and circumvallate papilla (CV). Tongue papillae except filiform papilla include taste buds. The papillae differ in taste sensitivities, likely due to differential expression of taste receptors. In this study, we evaluated differences in the expression levels of taste receptors in FU, FO and CV. Male DBA2 mice, 42-60 days old, were used in the study. Messenger RNAs were extracted from the murine epithelial tissues including FU, FO and CV. Cloned DNAs were synthesized by reverse transcription. Quantitative PCRs (qPCRs) were performed to determine mRNA expression levels of taste receptors. Results of qPCR revealed that the relative expression levels and patterns were different among FU, FO and CV. All three type 1 taste receptors were expressed FU, FO and CV at varying relative expression levels. All 35 kinds of type 2 taste receptors showed higher expression in FO and CV than in FU. Tas2r108 and Tas2r137 showed the two highest expression levels in all tested papillae. The differential expression levels and patterns of taste receptors among the three papillae could contribute to the different physiological sensitivities by tongue areas. Additional studies such as in situ hybridization or taste receptor cell activity recording is necessary to elucidate the functional relationship between expression levels of taste receptors and taste sensitivity.


Subject(s)
Animals , Clone Cells , DNA , Humans , In Situ Hybridization , Male , Mice , Mice, Inbred DBA , Polymerase Chain Reaction , Reverse Transcription , RNA, Messenger , Taste Buds , Tongue
7.
Rev. latinoam. enferm. (Online) ; 23(4): 571-577, July-Aug. 2015. tab
Article in English | LILACS, BDENF | ID: lil-761703

ABSTRACT

AbstractObjective: to analyze the process of tissue repair in patients with venous ulcers using inelastic compression therapy (the Unna Boot), in comparison with the use of the elastic bandage.Method: a controlled randomized clinical trial in which the patients (n=18) were allocated to two groups, those who used the Unna Boot (group B) and those who used the elastic bandage (group A). The study's follow-up period was 13 weeks.Results: a significant reduction took place, at the level of 5%, in the area, in square centimeters, of the ulcers of group B (p<0.0001) throughout the treatment, and there was a tendency of group A for reduction in the area of the ulcer, in centimeters squared (p=0.06), only after the fifth week.Conclusion: the treatment with the Unna Boot presented better results in venous ulcers with areas over 10cm², and the elastic bandage with Petrolatum(r) gauze in venous ulcers below 10cm². Brazilian Clinical Trials Register: Trial (req: 195) and WHO UTN U1111-1122-5489.


ResumoObjetivo:analisar o processo de reparo tecidual de pacientes com úlcera venosa em uso da terapia compressiva inelástica (Bota de Unna), em comparação ao uso da bandagem elástica.Método:ensaio clínico controlado randomizado em que os pacientes (n=18) foram alocados em dois grupos, os que utilizavam a Bota de Unna (grupo B) e os que utilizavam a atadura elástica (grupo A). O tempo de seguimento da pesquisa foi de treze semanas.Resultados:ocorreu redução significativa, no nível de 5%, na área, em centímetros quadrados, das úlceras do grupo B (p<0,0001) ao longo de todo o tratamento, e tendência do grupo A à redução, na área da úlcera, em centímetros quadrados (p=0,06), apenas após a quinta semana.Conclusão:o tratamento com a Bota de Unna apresentou melhor resultado em úlceras venosas com áreas superiores a 10cm², e a atadura elástica com a gaze Petrolatum(r)em úlceras venosas inferiores a 10cm². Registro Brasileiro de Ensaios Clínicos: Trial (req: 195) e WHO UTN U1111-1122-5489.


ResumenObjetivo:analizar el proceso de reparación del tejido de pacientes con úlcera venosa que usan la terapia compresiva inelástica (Bota de Unna), en comparación con el uso del vendaje elástico.Método:ensayo clínico controlado aleatorio en que los pacientes (n=18) fueron designados en dos grupos, los que utilizaban la Bota de Unna (grupo B) y los que utilizaban el vendaje elástico (grupo A). El tiempo de duración de la investigación fue de trece semanas.Resultados:se constató reducción significativa, al nivel de 5%, en el área, en centímetros cuadrados, de las úlceras del grupo B (p<0,0001) a lo largo de todo el tratamiento; y tendencia del grupo A a la reducción, en el área de la úlcera, en centímetros cuadrados (p=0,06), solamente después de la quinta semana.Conclusión:el tratamiento con la Bota de Unna presentó mejor resultado en úlceras venosas con áreas superiores a 10cm², y el vendaje elástico con la gasa Petrolatum(r)en úlceras venosas inferiores a 10cm². Registro Brasileño de Ensayos Clínicos: Trial (req: 195) y WHO UTN U1111-1122-5489.


Subject(s)
Animals , Female , Mice , Graft vs Host Disease , Kidney Neoplasms , Lymphocyte Transfusion , Stem Cell Transplantation , Allografts , Cell Line, Tumor , Graft vs Host Disease/immunology , Graft vs Host Disease/pathology , Graft vs Host Disease/therapy , Kidney Neoplasms/immunology , Kidney Neoplasms/pathology , Kidney Neoplasms/therapy , Mice, Inbred BALB C , Mice, Inbred DBA , Neoplasm Metastasis
8.
Article in English | WPRIM | ID: wpr-223789

ABSTRACT

Eupatilin is the main active component of DA-9601, an extract from Artemisia. Recently, eupatilin was reported to have anti-inflammatory properties. We investigated the anti-arthritic effect of eupatilin in a murine arthritis model and human rheumatoid synoviocytes. DA-9601 was injected into collagen-induced arthritis (CIA) mice. Arthritis score was regularly evaluated. Mouse monocytes were differentiated into osteoclasts when eupatilin was added simultaneously. Osteoclasts were stained with tartrate-resistant acid phosphatase and then manually counted. Rheumatoid synoviocytes were stimulated with TNF-alpha and then treated with eupatilin, and the levels of IL-6 and IL-1beta mRNA expression in synoviocytes were measured by RT-PCR. Intraperitoneal injection of DA-9601 reduced arthritis scores in CIA mice. TNF-alpha treatment of synoviocytes increased the expression of IL-6 and IL-1beta mRNAs, which was inhibited by eupatilin. Eupatilin decreased the number of osteoclasts in a concentration dependent manner. These findings, showing that eupatilin and DA-9601 inhibited the expression of inflammatory cytokines and the differentiation of osteoclasts, suggest that eupatilin and DA-9601 is a candidate anti-inflammatory agent.


Subject(s)
Animals , Anti-Inflammatory Agents/pharmacology , Arthritis, Experimental/chemically induced , Arthritis, Rheumatoid/drug therapy , Cell Differentiation/drug effects , Cells, Cultured , Collagen Type II , Cytokines/biosynthesis , Disease Models, Animal , Drugs, Chinese Herbal/therapeutic use , Female , Flavonoids/pharmacology , Humans , Inflammation/drug therapy , Interleukin-1beta/genetics , Interleukin-6/genetics , Lymph Nodes/cytology , Mice , Mice, Inbred DBA , Monocytes/cytology , Osteoclasts/cytology , Plant Extracts/pharmacology , RNA, Messenger/biosynthesis , Synovial Membrane/cytology , T-Lymphocytes, Regulatory/cytology , Tumor Necrosis Factor-alpha/pharmacology
9.
ABCD arq. bras. cir. dig ; 28(1): 48-52, 2015. tab, graf
Article in English | LILACS | ID: lil-742757

ABSTRACT

BACKGROUND: Few studies evaluated the association between nutritional disorders, quality of life and weight loss in patients undergoing bariatric surgery. AIM: To identify nutritional changes in patients undergoing bariatric surgery and correlate them with weight loss, control of comorbidities and quality of life. METHOD: A prospective cohort, analytical and descriptive study involving 59 patients undergoing bariatric surgery was done. Data were collected preoperatively at three and six months postoperatively, evaluating nutritional aspects and outcomes using BAROS questionnaire. The data had a confidence interval of 95%. RESULTS: The majority of patients was composed of women, 47 (79.7%), with 55.9% of the series with BMI between 40 to 49.9 kg/m². In the sixth month after surgery scores of quality of life were significantly higher than preoperatively (p<0.05) and 27 (67.5 %) patients had comorbidities resolved, 48 (81.3 %) presented BAROS scores of very good or excellent. After three and six months of surgery 16 and 23 presented some nutritional disorder, respectively. There was no relationship between the loss of excess weight and quality of life among patients with or without nutritional disorders. CONCLUSION: Nutritional disorders are uncommon in the early postoperative period and, when present, have little or no influence on quality of life and loss of excess weight. .


RACIONAL: Poucos estudos avaliam a associação entre distúrbios nutricionais, qualidade de vida e perda de peso em pacientes submetidos à cirurgia bariátrica. OBJETIVO: Identificar alterações nutricionais em pacientes submetidos à cirurgia bariátrica e correlacioná-las com perda de peso, controle de comorbidades e qualidade de vida. MÉTODO: Estudo de coorte, prospectivo, analítico e descritivo envolvendo 59 pacientes submetidos à cirurgia bariátrica. Os dados foram coletados no pré-operatório e aos três e seis meses pós- operatórios, quantificando aspectos nutricionais e utilizando o Bariatric Analysis and Reporting Outcomes System (BAROS) como ferramenta de sucesso. Os dados usaram intervalo de confiança de 95%. RESULTADOS: O total de mulheres foi 47 (79,7%), sendo 55,9% com IMC entre 40-49,9 kg/m². No sexto mês depois da operação os escores de qualidade de vida foram significativamente maiores do que no pré-operatório (p<0,05) e 27 (67,5%) pacientes tinham todas comorbidades resolvidas, 48 (81,3%) apresentaram conceito BAROS muito bom ou excelente. Após três e seis meses 16 e 23 pacientes apresentaram algum distúrbio nutricional, respectivamente. Não houve relação entre a perda do excesso de peso e qualidade de vida entre pacientes com ou sem distúrbio nutricional. CONCLUSÃO: os distúrbios nutricionais são pouco frequentes no pós-operatório precoce e, quando presentes, têm pouca ou nenhuma influência na qualidade de vida e na perda do excesso de peso. .


Subject(s)
Animals , Humans , Mice , Antibodies, Monoclonal/immunology , Antigens, Tumor-Associated, Carbohydrate/immunology , Neoplasms/immunology , Polysaccharides/immunology , Antibody-Dependent Cell Cytotoxicity , Cell Line, Tumor , Epitopes , Fas Ligand Protein/analysis , Fas Ligand Protein/immunology , Glycosylation , Mice, Inbred BALB C , Mice, Inbred DBA , Mannose/analysis , Vaccination
10.
Braz. dent. j ; 25(6): 565-570, Nov-Dec/2014. graf
Article in English | LILACS | ID: lil-732255

ABSTRACT

The passive fit of implant-supported dentures is fundamental to the rehabilitation success due the absence of the periodontal ligament in osseointegrated implants. Many techniques to obtain passive fit have been reported in the literature, some inaccessible for the clinicians and dental laboratories. This case report presents a technique to fabricate fixed complete dentures aiming at obtain passive fit with reduced time and cost, but without demerit for the aesthetics, function and longevity. A 40-year-old woman was referred for treatment presenting some teeth in the maxilla and an edentulous mandible, reporting eating problems related to instability and little retention of the mandibular complete denture. Treatment based on the reverse planning was performed to guide the rehabilitation with a complete mandibular fixed complete denture and maxillary occlusal plane adjustment. The framework of the fixed complete denture was manufactured luting a cast metal bar above the prepared titanium cylinder abutments using resin cement. The aim of this technique was to obtain a fixed complete denture with passive fit presenting positive esthetic and functional outcomes after 2 years of follow-up.


A adaptação passiva de próteses implantossuportadas é fundamental para o sucesso da reabilitação devido à inexistência de ligamento periodontal em implantes osseointegrados. Inúmeras técnicas de confecção da infraestrutura destas próteses tem sido relatadas na literatura, algumas inacessíveis para os clínicos e laboratórios de prótese. Este relato de caso apresenta uma técnica para confecção de próteses totais fixas visando obtenção de adaptação passiva com tempo e custo reduzido, porém sem demérito à estética, função e longevidade. Uma paciente de 40 anos se apresentou para tratamento apresentando alguns dentes na maxila e mandíbula edêntula, relatando dificuldades na mastigação relacionados a instabilidade e falta de retenção da prótese total inferior. Foi realizado um planejamento reverso para orientar a reabilitação com prótese total mandibular fixa e adequação do plano oclusal da maxila. A infraestrutura da prótese total fixa foi confeccionada pela cimentação de uma barra metálica em cilindros de titânio preparados com cimento resinoso. O objetivo desta técnica foi obter uma prótese total fixa com adaptação passiva apresentando resultados positivos em termos de estética e função após 2 anos de acompanhamento.


Subject(s)
Animals , Female , Male , Mice , Adenocarcinoma/drug therapy , Antineoplastic Agents/therapeutic use , Colonic Neoplasms/drug therapy , Floxuridine/therapeutic use , Fluorouracil/therapeutic use , Adenocarcinoma/pathology , Adenocarcinoma/radiotherapy , Bone Marrow/pathology , Combined Modality Therapy , Colonic Neoplasms/pathology , Colonic Neoplasms/radiotherapy , Floxuridine/administration & dosage , Floxuridine/toxicity , Fluorouracil/administration & dosage , Fluorouracil/toxicity , Mice, Inbred BALB C , Mice, Inbred DBA , Spleen/pathology , Thymus Gland/pathology , Weight Gain
11.
Journal of Experimental Hematology ; (6): 1115-1119, 2014.
Article in Chinese | WPRIM | ID: wpr-302337

ABSTRACT

This study was aimed to investigate the effect of different irradiation doses on the establishment of murine cGVHD model after MHC matched spleen stem cell transplantation. The male mouse BALB/c(H)-2d was totally irradiated with different radiation dose of (60)Co (TBI), then was infused with the same number of splenocytes from MHC matched DBA/2 male mice. After transplantation, the bodyweight, general appearance, hair changes, survival time and pathological damage were observed. The results indicated that compared to the control group (0 Gy) and the 7.0 Gy group, the mice irradiated with 7.5 Gy and 8.0 Gy showed cGVHD symptoms and obvious pathological damage. At the end of experiments (60 d after transplantation), all mice irradiated by 7.5 Gy survived while only 60% animals survived in the 8.0 Gy group. It is concluded that under infusion of 10(8) MHC matched splenocytes per mouse, 7.5 Gy irradiation is appropriate to efficiently establish cGVHD model. This study laid an important foundation for further studying the pathogenesis, biological characteristics, and intervention factors of cGVHD.


Subject(s)
Animals , Disease Models, Animal , Graft Survival , Radiation Effects , Graft vs Host Disease , Male , Mice , Mice, Inbred BALB C , Mice, Inbred DBA , Radiation Dosage , Spleen , Cell Biology , Stem Cell Transplantation , Transplantation Conditioning , Transplantation, Homologous , Whole-Body Irradiation
12.
Article in Chinese | WPRIM | ID: wpr-243476

ABSTRACT

<p><b>OBJECTIVE</b>We used an animal model of collagen-induced arthritis (CIA) to study changes and roles of tyrosine hydroxylase (TH) expressed by CD4+ T cell subsets, and then explore the relationship between CD4+ T cell subset-derived catecholamines and inflammatory responses in CIA.</p><p><b>METHODS</b>Thirty-six male DBA/1 mice were randomly divided into control group, CIA model group (day 35) and CIA model group (day 55) (n = 12). CIA model was induced by type II collagen (CII) in DBA/1 mice. On the 35th and 55th day following primary immunization, the joints of the mice were observed for clinical score of swelling and the level of anti-CII IgG antibody in serum was examined. Expression of specific transcription factors and cytokines of Th1, Th17, Th2 and Treg cells and TH in mesenteric lymph nodes was measured by means of Western blot. The changes of TH expressed by CD4+ T cell subsets in mesenteric lymph nodes were determined by flow cytometry.</p><p><b>RESULTS</b>Clinical score and anti-CII antibody level increased in CIA compared with that in intact mice. Specific transcription factors and cytokines expressed by Th1 and Th17 cells were upregulated and cytokines expressed by Th2 and Treg cells were downregulated in mesenteric lymph nodes in CIA mice. Expression of TH was upregulated and the increased expression of TH in CD4+ T cells was attributed to Th1 and Th17 cells in mesenteric lymph nodes of CIA.</p><p><b>CONCLUSION</b>The increase in catecholamines from CD4+ T cell subsets in mesenteric lymph nodes of CIA may be related to inflammatory alleviation in CIA progression.</p>


Subject(s)
Animals , Arthritis, Experimental , Allergy and Immunology , CD4-Positive T-Lymphocytes , Collagen Type II , Cytokines , Metabolism , Disease Models, Animal , Lymph Nodes , Allergy and Immunology , Male , Mice , Mice, Inbred DBA , T-Lymphocyte Subsets , Allergy and Immunology , Transcription Factors , Metabolism , Tyrosine 3-Monooxygenase , Metabolism
13.
Article in English | WPRIM | ID: wpr-223714

ABSTRACT

Interleukin (IL)-27 is a novel cytokine of the IL-6/IL-12 family that has been reported to be involved in the pathogenesis of autoimmune diseases and has a pivotal role as both a pro- and anti-inflammatory cytokine. We investigated the in vivo effects of IL-27 on arthritis severity in a murine collagen-induced arthritis (CIA) model and its mechanism of action regarding control of regulatory T (Tregs) and IL-17-producing T helper 17 (Th17) cells. IL-27-Fc-treated CIA mice showed a lower severity of arthritis. IL-17 expression in the spleens was significantly decreased in IL-27-Fc-treated CIA mice compared with that in the CIA model. The Th17 population was decreased in the spleens of IL-27-Fc-treated CIA mice, whereas the CD4+CD25+Foxp3+ Treg population increased. In vitro studies revealed that IL-27 inhibited IL-17 production in murine CD4+ T cells, and the effect was associated with retinoic acid-related orphan receptor gammaT and signal transducer and activator of transcription 3 inhibition. In contrast, fluorescein isothiocyanate-labeled forkhead box P3 (Foxp3) and IL-10 were profoundly augmented by IL-27 treatment. Regarding the suppressive capacity of Treg cells, the proportions of CTLA-4+ (cytotoxic T-lymphocyte antigen 4), PD-1+ (programmed cell death protein 1) and GITR+ (glucocorticoid-induced tumor necrosis factor receptor) Tregs increased in the spleens of IL-27-Fc-treated CIA mice. Furthermore, in vitro differentiated Treg cells with IL-27 exerted a more suppressive capacity on T-cell proliferation. We found that IL-27 acts as a reciprocal regulator of the Th17 and Treg populations in CD4+ cells isolated from healthy human peripheral blood mononuclear cells (PBMCs), as well as from humans with rheumatoid arthritis (RA) PBMCs. Our study suggests that IL-27 has the potential to ameliorate overwhelming inflammation in patients with RA through a reciprocal regulation of Th17 and Treg cells.


Subject(s)
Animals , Arthritis, Experimental/drug therapy , Cells, Cultured , Humans , Interleukins/immunology , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , T-Lymphocytes, Regulatory/immunology , Th17 Cells/immunology
14.
Article in Chinese | WPRIM | ID: wpr-235397

ABSTRACT

<p><b>OBJECTIVE</b>To show the involvement of lymphocyte-derived catecholamines in the pathogenesis of rheumatoid arthritis (RA), we investigated the change in expression of tyrosine hydroxylase (TH), a rate-limiting enzyme of catecholamine synthesis, by CD4+ T lymphocytes in lymphoid tissues of DBA/1 mice with collagen-induced arthritis (CIA).</p><p><b>METHODS</b>CIA model was induced by chicken type II collagen in DBA/1 mice. The joints of the mice were observed for clinical score of swelling on and after the 22nd day of primary immunization. Pathological changes of ankles were examined by staining of tissue sections with hematoxylin and eosin on the 35th and 55th day following primary immunization. Immunofluorescent histochemistry was used to identify the number of TH-positive, CD4-positive, and double-labeled cells in the mesenteric lymph nodes and the spleen.</p><p><b>RESULTS</b>Paw-swelling onset was on days 29 - 32 after the first immunization in DBA/1 mice. Clinical score for swelling of the paws reached peak on day 46 after the first immunization. Compared with the ankles of intact or vehicle mice, the joints of CIA mice had these characteristics: increased inflammatory cells in the synovial tissues, proliferated synoviocytes in the multilayers, narrowed articular space, and destructed articular cartilages. Simultaneously, the number of TH-positive, CD4-positive, and double-labeled cells in the mesenteric lymph nodes and the spleen was significantly increased on days 35 and 55 following the first immunization. Between day 35 and day 55 post-immunization, there was no significant difference in the number of these positive cells.</p><p><b>CONCLUSION</b>CD4+ T lymphocytes up-regulate TH expression in the process of CIA and therefore, it is suggested that endogenous catecholamines of lymphocytes involve in the pathogenesis of RA.</p>


Subject(s)
Animals , Arthritis, Experimental , Metabolism , Arthritis, Rheumatoid , CD4-Positive T-Lymphocytes , Metabolism , Lymphoid Tissue , Metabolism , Male , Mice , Mice, Inbred DBA , Tyrosine 3-Monooxygenase , Metabolism
15.
Article in Chinese | WPRIM | ID: wpr-286016

ABSTRACT

<p><b>OBJECTIVE</b>To establish a simple and highly effective isolation and culture system of mouse spermatogonial stem cells(SSCs)and detect the expression of stem cell-related markers in the isolated cells.</p><p><b>METHODS</b>The structures of seminiferous tubules of neonatal(6-8 days of age)and adult(26-28 weeks)DBA/2 mice were compared using histochemical examination. Testes of neonatal mice were selected for preparing primary cells. The digestive efficiency of different enzymes was compared. SSCs were isolated according to the different binding abilities of testicle somatic cells and SSCs to gelatin matrix. The effects of different base culture media such as StemPro34 and α-MEM,gelatin,and serum on the SSCs binding activity and growth were studied. The cell morphology was observed during the culture process. Immunofluorescence was used to detect the expression of SSCs and cancer stem cells(CSCs)-related markers in SSCs.</p><p><b>RESULTS</b>The content of SSCs in the testes of neonatal mice was relatively higher than that in adult mice. Trypsin showed the highest digestive efficiency. In StemPro34 supplemented with 1% fetal bovine serum and on the gelatin matrix,testicular somatic cells could bind with the plate efficiently. Spermatogonial cells grew well when using mitomycin C-treated testicular somatic cells as feeder cells and showed typical characteristic of SSCs. After 13 days of culture,spermatogonial cells formed cell clusters. Immunofluorescence assay showed that SSCs markers glial cell line-derived neurotrophic factor(GDNF)family receptor α1(GFRα1)and VASA protein were highly expressed in the cell clusters. CSCs marker CD44 was expressed in the As,Apr,Aal and the inner cells of the cell clusters,while seldom expressed in the somatic cells.</p><p><b>CONCLUSIONS</b>An isolation and culture system of SSCs derived from DBA/2 mice was established. CD44 is highly expressed in the early stage of spermatogonial cell development.</p>


Subject(s)
Animals , Biomarkers , Metabolism , Cell Culture Techniques , Cell Differentiation , Cells, Cultured , Glial Cell Line-Derived Neurotrophic Factor , Metabolism , Hyaluronan Receptors , Metabolism , Male , Mice , Mice, Inbred DBA , Spermatogonia , Cell Biology , Stem Cells , Cell Biology
16.
Article in English | WPRIM | ID: wpr-347113

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of sodium copper chlorophyllin (SCC) on the proliferation, differentiation and immunomodulatory function of mesenchymal stem cells (MSCs) from mice with aplastic anemia.</p><p><b>METHODS</b>A mouse model of aplastic anemia was established by exposure of BALB/c mice to sublethal doses of 5.0 Gy Co60 γ radiation, followed by transplantation of 2×10(6) lymph node cells from DBA/2 donor mice within 4 h after radiation. Aplastic anemic BALB/c mice were randomly divided into six groups: the treated groups, which received 25, 50, or 100 mg/kg/day SCC, respectively; a positive control group treated with cyclosporine A (CsA); and an untreated model control group (model group); while, the non-irradiated mice as the normal control group. SCC or CsA were administered by gastrogavage for 20 days, starting on day 4 after irradiation. Peripheral blood cells were counted and colony-forming fibroblasts (CFU-F) in the bone marrow were assayed. The ability of MSCs to form calcium nodes after culture in osteoinductive medium was also observed. The immunosuppressive effect of MSCs on T lymphocytes was analyzed by enzyme-linked immunosorbent assay and flow cytometry, to evaluate the efficacy of SCC in mice with aplastic anemia.</p><p><b>RESULTS</b>Peripheral blood white cell and platelet counts were increased by medium and high SCC doses, compared with the untreated control. CFU-Fs were also increased compared with the untreated control, and the numbers of calcium nodes in MSCs in osteoinductive medium were elevated in response to SCC treatment. The percentage of Forkhead box protein 3 (FOXP3(+)) T cells was increased in T cell-MSC cocultures, and the cytokine transforming growth factor β1 was up-regulated in SCC-treated groups.</p><p><b>CONCLUSION</b>The results of this study suggest that SCC not only promotes the proliferation and differentiation of MSCs, but also improves their immunoregulatory capacity in mice with aplastic anemia.</p>


Subject(s)
Anemia, Aplastic , Blood , Pathology , Therapeutics , Animals , Anthraquinones , Metabolism , Biomarkers , Metabolism , Bone Marrow Cells , Pathology , Calcium , Metabolism , Cell Differentiation , Cell Proliferation , Chlorophyllides , Pharmacology , Colony-Forming Units Assay , Female , Immunosuppression , Leukocyte Count , Male , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Cell Biology , Metabolism , Mice , Mice, Inbred BALB C , Mice, Inbred DBA , Osteoblasts , Pathology , Platelet Count , T-Lymphocytes
17.
Article in English | WPRIM | ID: wpr-59930

ABSTRACT

This study examined the effects of calcium (Ca) gluconate on collagen-induced DBA mouse rheumatoid arthritis (CIA). A single daily dose of 200, 100 or 50 mg/kg Ca gluconate was administered orally to male DBA/1J mice for 40 days after initial collagen immunization. To ascertain the effects administering the collagen booster, CIA-related features (including body weight, poly-arthritis, knee and paw thickness, and paw weight increase) were measured from histopathological changes in the spleen, left popliteal lymph node, third digit and the knee joint regions. CIA-related bone and cartilage damage improved significantly in the Ca gluconate- administered CIA mice. Additionally, myeloperoxidase (MPO) levels in the paw were reduced in Ca gluconate-treated CIA mice compared to CIA control groups. The level of malondialdehyde (MDA), an indicator of oxidative stress, decreased in a dose-dependent manner in the Ca gluconate group. Finally, the production of IL-6 and TNF-alpha, involved in rheumatoid arthritis pathogenesis, were suppressed by treatment with Ca gluconate. Taken together, these results suggest that Ca gluconate is a promising candidate anti-rheumatoid arthritis agent, exerting anti-inflammatory, anti-oxidative and immunomodulatory effects in CIA mice.


Subject(s)
Animals , Arthritis , Arthritis, Rheumatoid , Body Weight , Calcium Gluconate , Calcium , Cartilage , Collagen , Humans , Immunization , Immunomodulation , Interleukin-6 , Knee , Knee Joint , Lymph Nodes , Male , Malondialdehyde , Mice , Mice, Inbred DBA , Oxidative Stress , Peroxidase , Spleen , Tumor Necrosis Factor-alpha , Water
18.
Acta Pharmaceutica Sinica ; (12): 1210-1218, 2012.
Article in Chinese | WPRIM | ID: wpr-274675

ABSTRACT

In order to increase the plasma half-life and tissue specificity of IL-1 receptor antagonist, a recombinant fusion protein IL-1Ra-HSA, linked by a rigid peptide linker PAPAP, was engineered and expressed by the Pichia pastoris host cells. The fusion protein was secreted to the host cells culture, identified by Western blot, and purified by affinity chromatography. This was followed by a further examination of its bioactivity and pharmacokinetics. Our results demonstrated that the fusion protein retained the antagonist activity of IL-1Ra, capable of binding specifically to the IL-1 receptor on human melanoma A375.S2 cells, and inhibits the cytolytic effect of IL-1beta to A375.S2 cells. Albumin fusion dramatically extended the half-life of IL-1Ra and resulted in a specific accumulation of IL-1Ra in the arthritic paws and a lower distribution of IL-1Ra in other organs such as liver, kidney, spleen and lung in mice with collagen-induced arthritis. The findings reported herein indicate that the fusion protein is likely to have greater clinical applications in areas such as the treatment of rheumatoid arthritis.


Subject(s)
Animals , Apoptosis , Arthritis, Experimental , Metabolism , Cell Line, Tumor , Forelimb , Metabolism , Half-Life , Humans , Interleukin 1 Receptor Antagonist Protein , Genetics , Metabolism , Pharmacokinetics , Pharmacology , Interleukin-1beta , Toxicity , Male , Melanoma , Pathology , Mice , Mice, Inbred DBA , Pichia , Genetics , Metabolism , Plasmids , Recombinant Fusion Proteins , Genetics , Metabolism , Pharmacokinetics , Pharmacology , Serum Albumin , Genetics , Metabolism , Pharmacokinetics , Pharmacology , Tissue Distribution
19.
Chinese Journal of Virology ; (6): 103-107, 2012.
Article in Chinese | WPRIM | ID: wpr-354763

ABSTRACT

To explore the infectivity characteristics and susceptibility of Sendai strain Tianjin in 129Sv, DBA/2, Kunming and BALB/c mice and determine the optimal small rodent animal model for strain Tianjin research, the Sendai strain Tianjin was propagated for 72h in 9-11 day-old chicken embryos, the allantoic fluids were then harvested and the virus titer (1:1280) was detected by hemagglutination assay. Four different kinds of mice were intranasally inoculated with 5 microl and the diluted 30 microl virus solution. The weight loss of mice was monitored for 12 consecutive days and the survival rate was observed. Kunming and BALB/c mice were sacrificed on the first day prior to infection and on the fourth and seventh days post infection of the diluted 30 microl Sendai strain Tianjin. Their left lobes of lung were fixed with 4% formalin for histopathologic examination. The maximum percentage of average weight loss of 129Sv, DBA/2 were 13.0%, 4.7% with 100% survival rate when 129Sv, DBA/2, Kunming and BALB/c were inoculated with 5 microl virus solution, while the mice were inoculated with diluted 30 microl virus solution, the maximum percentage of average weight loss reached 21.7%, 30.3%, 16.7% and 9.7% with the survival rate of 20%, 0%, 80% and 100%. Lung infections of mice Kunming on the fourth and seventh day post infection were more severe than that of BALB/c, showing a large number of inflammatory cell exudation and thickening of the submucosa. It suggested that DBA/2 was the most susceptible to the infection of strain Tianjin. The mice susceptibility ranked as DBA/2>129Sv>Kunming>BALB/c. Mice DBA/2 and 129Sv could be used as the optimal small rodent animal models in the research of pathogenicity and vaccine of Sendai strain Tianjin.


Subject(s)
Animals , Disease Susceptibility , Female , Humans , Lung , Pathology , Virology , Mice , Mice, Inbred BALB C , Mice, Inbred DBA , Respirovirus Infections , Virology , Sendai virus , Classification , Physiology
20.
Article in English | WPRIM | ID: wpr-149759

ABSTRACT

IL-17-producing CD4+ T cells (Th17) play important functions in autoimmune diseases and allograft rejection of solid organs. We examined the effects of IL 17 and its mechanism of action on arthritis in a murine collagen-induced arthritis (CIA) model using bone marrow transplantation (BMT) system. DBA/1J mice were administered a lethal radiation dose and then rescued with bone marrow derived from either wild-type (WT) or IL-17-/- mice on C57BL/6 background mice. CIA was induced after the bone marrow transplant, and disease progression was characterized. DBA/1J mice with CIA that received IL-17-/- donor bone marrow showed potently inhibited development and severity of clinical arthritis as compared with CIA mice that received WT bone marrow. Reduced secretion of the pro-inflammatory cytokines tumor necrosis factor-alpha, IL-1beta, and IL-6, and collagen-specific T cell responses were observed in mice that received IL-17-/- bone marrow. IL-17 blockade also inhibited effector T cell proliferation by reciprocally regulating the Treg/Th17 ratio. IL-17 blockade prevented joint destruction in mice with CIA. These findings suggest that CIA with BMT is a viable method of immunological manipulation and that IL-17 deficiency suppresses severe joint destruction and inflammation in CIA mice. There may be clinical benefits in blocking IL-17 and BMT in the treatment of rheumatoid arthritis.


Subject(s)
Animals , Antigens, Differentiation/metabolism , Arthritis, Experimental/pathology , Bone Marrow Transplantation , Cell Differentiation , Cell Proliferation , Cells, Cultured , Collagen Type II , Cytokines/metabolism , Humans , Interleukin-17/deficiency , Joints/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Mice, Knockout , Osteoclasts/metabolism , Signal Transduction , T-Lymphocytes/metabolism , Transplantation, Homologous
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