RESUMO
OBJECTIVE: Ageing and aberrant biomechanical stimulation are two major risk factors for osteoarthritis (OA). One of the main characteristics of aged cartilage is cellular senescence. One of the main characteristics of osteoarthritic joints is cartilage degeneration. The cells in the temporomandibular joint (TMJ) cartilage are zonally arranged. The deep zone cells are differentiated from the superficial zone cells (SZCs). The purpose of the present study was to investigate whether degenerative shear stress (SS) stimulates the senescence programme in TMJ SZCs, and to determine which miRNA is involved in this process. METHOD: SZCs were isolated from the TMJ condyles of 3-week-old rats and treated with continuous passaging or SS. RNA sequencing was conducted to identify miRNA(s) that overlap with those involved in the replication senescence process and the SS-induced degeneration programme. Unilateral anterior crossbite (UAC), which is TMJ-OA inducible, was applied to 2-month-old and 12-month-old mice for 3 weeks. The effect of TMJ local injection of agomiR-708-5p was evaluated histologically. RESULTS: Both replication and SS treatment induced SZC senescence. miR-708-5p was identified. Knocking down miR-708-5p in SS-treated SZCs led to more severe senescence by alleviating the inhibitory impact of miR-708-5p on the TLR4/NF-κB pathway. miR-708-5p expression in mouse TMJ cartilage decreased with age. UAC induced more severe osteoarthritic cartilage lesions in 12-month-old mice than in 2-month-old mice. Injection of agomiR-708-5p suppressed UAC-induced osteoarthritic cartilage lesions. CONCLUSIONS: Age-related miR-708-5p deficiency is involved in the mechanically stimulated OA process. Intra-articular administration of agomiR-708-5p is a promising new strategy for OA treatment.
Assuntos
Condrócitos , Côndilo Mandibular , MicroRNAs , NF-kappa B , Receptor 4 Toll-Like , Animais , Feminino , Camundongos , Ratos , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Senescência Celular/genética , Condrócitos/metabolismo , Côndilo Mandibular/patologia , Camundongos Endogâmicos C57BL , MicroRNAs/genética , NF-kappa B/metabolismo , Osteoartrite/genética , Osteoartrite/metabolismo , Osteoartrite/patologia , Ratos Sprague-Dawley , Transdução de Sinais , Articulação Temporomandibular/patologia , Articulação Temporomandibular/metabolismo , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismoRESUMO
BACKGROUND: Gorham's disease (GSD) is a rare osteolytic disease with unclear etiology, and no known prevention or effective treatment. Here we report a new surgical treatment for a case of GSD in September 2017. CASE PRESENTATION: We report GSD in a 52-year-old woman. She had disappearance of her humeral head and a defect of the glenoid bone in her left shoulder joint, which were serious obstacles to joint function. We used an autologous iliac bone graft to repair the glenoid bone defect and a reverse total shoulder arthroplasty. After surgery, humeral osteolysis did not continue, and her shoulder function recovered well. CONCLUSIONS: This case suggests that autologous bone grafting can still be used to treat GSD despite it being an osteolytic disease. The successful treatment suggests that this method could be used for GSD in other bones.
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Artroplastia de Substituição , Transplante Ósseo/métodos , Ílio/transplante , Osteólise Essencial/cirurgia , Articulação do Ombro/cirurgia , Artroplastia de Substituição/instrumentação , Autoenxertos , Fenômenos Biomecânicos , Biópsia , Feminino , Humanos , Imageamento por Ressonância Magnética , Pessoa de Meia-Idade , Osteólise Essencial/diagnóstico por imagem , Osteólise Essencial/fisiopatologia , Amplitude de Movimento Articular , Recuperação de Função Fisiológica , Articulação do Ombro/diagnóstico por imagem , Articulação do Ombro/fisiopatologia , Prótese de Ombro , Tomografia Computadorizada por Raios X , Resultado do TratamentoRESUMO
Calcium phosphate cement (CPC) porous scaffold is widely used as a suitable bone substitute to repair bone defect, but the optimal pore size is unclear yet. The current study aimed to evaluate the effect of different pore sizes on the processing of bone formation in repairing segmental bone defect of rabbits using CPC porous scaffolds. Three kinds of CPC porous scaffolds with 5 mm diameters and 12 mm length were prepared with the same porosity but different pore sizes (Group A: 200-300 µm, Group B: 300-450 µm, Group C: 450-600 µm, respectively). Twelve millimeter segmental bone defects were created in the middle of the radius bone and filled with different kinds of CPC cylindrical scaffolds. After 4, 12, and 24 weeks, alkaline phosphatase (ALP), histological assessment, and mechanical properties evaluation were performed in all three groups. After 4 weeks, ALP activity increased in all groups but was highest in Group A with smallest pore size. The new bone formation within the scaffolds was not obvious in all groups. After 12 weeks, the new bone formation within the scaffolds was obvious in each group and highest in Group A. At 24 weeks, no significant difference in new bone formation was observed among different groups. Besides the osteoconductive effect, Group A with smallest pore size also had the best mechanical properties in vivo at 12 weeks. We demonstrate that pore size has a significant effect on the osteoconductivity and mechanical properties of calcium phosphate cement porous scaffold in vivo. Small pore size favors the bone formation in the early stage and may be more suitable for repairing segmental bone defect in vivo.
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Cimentos Ósseos/uso terapêutico , Regeneração Óssea , Substitutos Ósseos/uso terapêutico , Fosfatos de Cálcio/uso terapêutico , Osteogênese , Fosfatase Alcalina/metabolismo , Animais , Fenômenos Biomecânicos , Cimentos Ósseos/química , Regeneração Óssea/efeitos dos fármacos , Substitutos Ósseos/química , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/lesões , Fosfatos de Cálcio/química , Masculino , Osteogênese/efeitos dos fármacos , Porosidade , Coelhos , Alicerces Teciduais/químicaRESUMO
As the predominant group inhabiting the human gastrointestinal tract, bifidobacteria play a vital role in human nutrition, therapeutics, and health by shaping and maintaining the gut ecosystem, reducing blood cholesterol, and promoting the supply of nutrients. The interaction between bacterial cells and human intestinal epithelial cell lines has been studied for decades in an attempt to understand the mechanisms of action. These studies, however, have been limited by lack of genomic and proteomic database to aid in achieving comprehensive understanding of these mechanisms at molecular levels. Microarray data (GSE: 74119) coupled with isobaric tags for relative and absolute quantitation (iTRAQ) were performed to detect differentially expressed genes and proteins in HT-29 cells after incubation with Bifidobacterium bifidum. Real-time quantitative PCR, gene ontology, and Kyoto Encyclopedia of Genes and Genomes analyses were further conducted for mRNA validation, functional annotation, and pathway identification, respectively. According to the results of microarray, 1,717 differentially expressed genes, including 1,693 upregulated and 24 downregulated genes, were selected and classified by the gene ontology database. The iTRAQ analysis identified 43 differentially expressed proteins, where 29 proteins were upregulated and 14 proteins were downregulated. Eighty-two candidate genes showing consistent differences with microarray and iTRAQ were further validated in HT-29 and Caco-2 cells by real-time quantitative PCR. Nine of the top genes showing interesting results with high confidence were further investigated in vivo in mice intestine samples. Integration of genomic and proteomic data provides an approach to identify candidate genes that are more likely to function in ubiquitin-mediated proteolysis, positive regulation of apoptosis, membrane proteins, and transferase catalysis. These findings might contribute to our understanding of molecular mechanisms regulating the interaction between probiotics and intestinal epithelial cell lines.
Assuntos
Bifidobacterium bifidum/crescimento & desenvolvimento , Células Epiteliais/metabolismo , Proteoma , Animais , Células CACO-2 , Bases de Dados Genéticas , Regulação para Baixo , Células Epiteliais/microbiologia , Microbioma Gastrointestinal , Ontologia Genética , Células HT29 , Humanos , Intestinos/citologia , Intestinos/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Probióticos , Proteômica , Reprodutibilidade dos Testes , Regulação para CimaRESUMO
In this study, we investigated the effect of administration of 5 strains of lactic acid bacteria (LAB) isolated from traditional Chinese sourdough on the flora balance of gastrointestinal tract of mice. We specifically measured Enterococcus, Enterobacter, Bacteroides, and Lactobacillus by plate count and real-time PCR methods, and α-glucosidase, lactate dehydrogenase, esterase, and aminopeptidase activities as indicative of metabolism of sugar, fat, and protein from LAB isolated from feces of mice in vitro. The results showed that administration of Lactobacillus acidophilus LAC0201 and Lactobacillus fermentum LFE0302 lowered the uricacid index of serum. Lactobacillus acidophilus LAC0201, L. fermentum LFE0302, as well as Lactobacillus curvatus LCU0401 administration resulted in a reduction in the opportunistic pathogens (i.e., Enterococcus and Enterobacter), meanwhile, administration of L. fermentum LFE0302 and Lactobacillus sp. ULA0104 resulted in an increase in the counts of Lactobacillus. Lactobacillus fermentum LFE0302 administration increased starch digestion of intestinal flora after 4wk of feeding and also resulted in increased α-glucosidase activity in the intestinal flora after 3wk of feeding. We found a similar trend in esterase activity after administration of L. acidophilus LAC0201 for 3wk. Hence, our study suggested that LAB from Chinese sourdough might be used as potential probiotics to strengthen the flora balance in gastrointestinal tract and positively change the metabolism of nutrients through bacterial enzyme activities.
Assuntos
Ácido Láctico/metabolismo , Lactobacillus/metabolismo , Animais , Carboidratos , Trato Gastrointestinal/metabolismo , Camundongos , ProbióticosRESUMO
Twelve isolates isolated from the gastrointestinal tracts of Gaotian villagers in China, who had a lifespan of 92 yr, were examined for their antioxidants using free radical scavenging activity and 2,2-diphenyl-1-picrylhydrazyl. Three strains (i.e., Lactobacillus mucosae LMU1001, and Lactobacillus plantarum LPL0902 and LPL0302) were selected as candidates to prepare yogurt for testing their antioxidants in a model of d-galactose-induced aging mice, with vitamin C as a positive control. The results showed that L. mucosae LMU1001 was the best strain, which had similar in vivo antioxidant activity as vitamin C. A significant increase was found in the activities of glutathione peroxidase in serum and total superoxide dismutase in the liver, and a decrease in the level of malondialdehyde in serum. Regarding mRNA expression level detected quantitatively by real-time PCR, we observed that L. mucosae LMU1001 significantly upregulated antioxidant genes (i.e., MT1A and MT1M in HT-29 and Caco-2) and those genes (i.e., MT1, MT2, GPx1, and GPx2) in the intestinal tract of the model mice. Hence, this strain could be considered as a potential probiotic lactic acid bacterium for improving antioxidant levels in functional foods.
Assuntos
Envelhecimento , Antioxidantes/análise , Galactose/farmacologia , Intestinos/microbiologia , Lactobacillus/fisiologia , Idoso de 80 Anos ou mais , Envelhecimento/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Células CACO-2 , China , Alimento Funcional , Glutationa Peroxidase/sangue , Humanos , Lactobacillus/isolamento & purificação , Lactobacillus plantarum/fisiologia , Fígado/enzimologia , Longevidade , Malondialdeído/sangue , Camundongos , Modelos Animais , Probióticos , Superóxido Dismutase/metabolismoRESUMO
Autophagy is a cell protective mechanism for maintaining cellular homeostasis. The present study aimed to investigate whether autophagy is enhanced in the biomechanically induced degenerative cartilage of the temporomandibular joint (TMJ) and the potential role of mitogen-activated protein kinase kinase kinase kinase 3 (MAP4K3) and mammalian Target of rapamycin (mTOR) in this observation. To induce degenerative changes in the TMJs, rats were subjected to biomechanical dental stimulation by moving 4 molars away from their original position as we previously reported. The ultrastructure of autophagosome was observed by transmission electron microscopy. The number of lysosomes was analyzed by flow cytometry. The expression levels of Beclin1 and LC3 and the involvement of MAP4K3 activity were detected by immunohistochemistry, real-time PCR and western blot. The activity of the mTOR pathway indicated by p-mTOR and p-p70S6 K was assayed by western blot. TMJ degeneration, characterized by irregular cell arrangement and cell-free area, was induced in the experimental groups. Under transmission electron microscopy, we observed the presence of autophagosomes, small patches of condensed chromatin, abundant rough endoplasmic reticulum and Golgi apparatus. The number of lysosomes and the expression levels of Beclin1 and LC3 increased, while the activity of mTOR and the expression level of MAP4K3 decreased in the experimental groups. Cartilage in TMJ which was induced to be degenerative biomechanically exhibited autophagy accompanied by reduced mTOR and MAP4K3 activity.
Assuntos
Autofagia , Cartilagem/fisiologia , Condrócitos/fisiologia , Articulação Temporomandibular/fisiologia , Técnicas de Movimentação Dentária , Animais , Proteínas Reguladoras de Apoptose/biossíntese , Proteína Beclina-1 , Sobrevivência Celular , Feminino , Lisossomos , Proteínas Associadas aos Microtúbulos/biossíntese , Proteínas Serina-Treonina Quinases/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , Transdução de Sinais , Estresse Mecânico , Serina-Treonina Quinases TOR/metabolismoRESUMO
BACKGROUND: It is a challenge to perform a joint-preserving resection for patients with bone sarcomas in the proximal humerus. We determined whether osteotomy under navigation guidance made joint-saving resection possible for juxtaarticular humeral sarcomas while adhering to oncological principles. METHOD: Between January 2008 and July 2010, joint-preserving surgeries were performed on six patients with proximal humeral sarcomas under navigation guidance. Five tumors extended to, and one extended beyond, the epiphyseal line. Planned osteotomy under image-guided navigation was employed to achieve a clear surgical margin while preserving the humeral head and rotator cuff. All tumors were removed en bloc and intercalary defects were reconstructed by a combination of allograft and vascularized fibula graft. All specimens were examined for resection margin. Patients were followed up for an average of 19.1 months. RESULTS: The entire glenohumeral joint was preserved in five patients and part of the humeral head was saved in one patient. Clear surgical margin was obtained in all specimens. The minimum closest distance between the osteotomy line and tumor edge was 7 mm. No patient experienced local recurrence. One patient developed lung metastasis and was alive with disease. The mean Musculoskeletal Tumor Society (MSTS) 93 score was 92.1%. All reconstruction was in situ at final follow-up. CONCLUSION: With careful patient selection, image navigation-assisted surgery made it possible to excise the bone exactly as seen in orientation in magnetic resonance imaging (MRI) image, yielding a clear margin and preserving all or part of the humeral head in limb salvage procedures for patients with juxtaarticular bone sarcomas in proximal humerus. LEVEL OF EVIDENCE: Therapeutic study; Level IV.
Assuntos
Neoplasias Ósseas/cirurgia , Úmero/cirurgia , Salvamento de Membro/métodos , Sarcoma/cirurgia , Cirurgia Assistida por Computador , Adolescente , Adulto , Criança , Feminino , Fíbula/transplante , Seguimentos , Humanos , Imageamento por Ressonância Magnética , Masculino , Osteotomia/métodos , Seleção de Pacientes , Tomografia Computadorizada por Raios X , Adulto JovemRESUMO
BACKGROUND: Osteoarthritis (OA) is an inflammatory joint disorder with high incidence rates. Long non-coding RNAs (LncRNAs) influence OA development. OBJECTIVES: In this research, we attempt to figure out the functions of lncRNA BLACAT1 in human articular chondrocyte (HAC) apoptosis and extracellular matrix (ECM) degradation in OA. METHODS: Interleukin (IL)-1ß was employed to induce HAC damage. Cell viability and apoptosis were detected, with expression patterns of lncRNA BLACAT1, miR-149-5p, and HMGCR, and levels of Caspase-3, Caspase-9, BAX, Bcl-2, COL2A1, and SOX9 determined. Then, lncRNA BLACAT1 was silenced in IL-1ß-treated HACs to analyze its role in HAC damage. The target relations of lncRNA BLACAT1 and miR-149-5p and miR-149-5p and HMGCR were verified. In addition, combined experiments were performed as a miR-149-5p inhibitor or HMGCR overexpression was injected into cells with lncRNA BLACAT1 silencing. RESULTS: In IL-1ß-treated HACs, lncRNA BLACAT1 and HMGCR were overexpressed while miR- 149-5p was poorly expressed, along with reduced cell viability, enhanced apoptosis, elevated Caspase-3 and Caspase-9 activities, increased BAX level, decreased Bcl-2 level, and declined levels of COL2A1 and SOX9, which were reversed by lncRNA BLACAT1 silencing. LncRNA BLACAT1 targeted miR-149-5p, and miR-149-5p targeted HMGCR. miR-149-5p knockout or HMGCR overexpression annulled the inhibitory role of lncRNA BLACAT1 silencing in HAC apoptosis and ECM degradation. CONCLUSION: LncRNA BLACAT1 was overexpressed in IL-1ß-treated HACs, and the lncRNA BLACAT1/miR-149-5p/HMGCR ceRNA network promoted HAC apoptosis and ECM degradation.
Assuntos
Osteoartrite , RNA Longo não Codificante/metabolismo , Apoptose , Caspase 3/metabolismo , Caspase 9/metabolismo , Condrócitos/metabolismo , Matriz Extracelular/metabolismo , Humanos , Hidroximetilglutaril-CoA Redutases/metabolismo , Interleucina-1beta/metabolismo , Interleucina-1beta/farmacologia , Redes e Vias Metabólicas , MicroRNAs/genética , MicroRNAs/metabolismo , Osteoartrite/genética , Osteoartrite/metabolismo , RNA Longo não Codificante/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
OBJECTIVES: The aim of this study was to investigate the impact of unified discharge standards on the clinical efficacy and prognosis of hemiarthroplasty in elderly patients with hip fractures. METHODS: Retrospective study of 158 elderly patients with hip fractures who underwent artificial femoral head replacement in our hospital from March 2016 to July 2019 were enrolled. According to the unified discharge standards, patients were divided into the observation (65 cases who met discharge criteria) and control group (93 cases who failed to meet all discharge criteria). Histopathological feature, operation status, postoperative Harris Hip score, therapeutic outcome, postoperative complications, readmission and mortality rate were compared between the two groups. RESULTS: Surgery duration and intraoperative blood loss exhibited no difference between the two groups, while transfusion volume and length of hospital stay were significantly increased in the observation group. There was no significant difference in the Harris Hip score between the two groups 12 months postoperatively. The incidence of postoperative complications during the follow-up period was notably lower in the observation group. In addition, the three-month readmission rate and one-year mortality rate were significantly lower in the observation group. CONCLUSIONS: For elderly patients with hip fractures undergoing artificial femoral head replacement, the incidence of postoperative complications and postoperative readmission/mortality rate could be reduced through the establishment of unified discharge standards, which should be used in future clinical practice.
RESUMO
In the adult skeleton, the bone remodeling process involves a dynamic coordination between osteoblasts and osteoclasts, which is disrupted in diseases with high bone turnover rates and dysregulated transforming growth factor beta 1 (TGF-ß1). However, little is known about how TGF-ß1 signaling mediates bone resorption. Here, we described a pedigree with a heterozygous variant in TGF-ß1 (R218C) that resulted in aberrant activation of TGF-ß1 through an activating mechanism that caused Camurati-Engelmann disease (CED). We showed that CED patients have high levels of active Rho GTPases and the migration-related proteins Integrin ß1 and Integrin ß3 in their peripheral blood. HEK293T cells transfected with a plasmid encoding this mutant expressed high levels of TGF-ß1 and active Rho GTPases. Furthermore, activation of Rho by TGF-ß1 increased osteoclast formation and bone resorption, with increased migration of pre-osteoclasts, as well as cytoskeletal remodeling of pre-osteoclasts and mature osteoclasts. Importantly, pharmacological inhibition of Rho GTPases effectively rescued hyperactive TGF-ß1-induced osteoclastogenesis in vitro. Overall, we propose that Rho GTPases mediate TGF-ß1-induced osteoclastogenesis and suggest that Rho-TGF-ß1 crosstalk is associated with high bone turnover in CED.
Assuntos
Reabsorção Óssea , Síndrome de Camurati-Engelmann , Adulto , Humanos , Fator de Crescimento Transformador beta1/farmacologia , Fator de Crescimento Transformador beta1/metabolismo , Proteínas rho de Ligação ao GTP/genética , Proteínas rho de Ligação ao GTP/metabolismo , Células HEK293 , Remodelação ÓsseaRESUMO
Honokiol is the main active constituent of Magnolia officinalis. With effective and longterm pharmacological functions of being antibacterial, antioxidative, antiinflammatory, antitumor, antispasmic, antianxiety and antiviral, Honokiol is clinically used in the treatment of acute enteritis and chronic gastritis. The aim of the present study was to observe the possible antieffects of honokiol on autophagy and apoptosis of osteosarcoma, and to investigate the role of the PI3K/Akt/mTOR signaling pathway in its anticancer effects. MTT assay was used to evaluate cell proliferation and Annexin Vfluorescein isothiocyanate/propidium iodide staining flow cytometry was used to analyze the apoptotic rate. The authors identified that honokiol could inhibit cell proliferation and induce the apoptotic rate of osteosarcoma cells. The expression level of Bcl2like protein 4, caspase3 and p53 protein expression were induced and cyclin D1 protein expression was suppressed in osteosarcoma cells by honokiol. Autophagyassociated LC3II protein expression level was promoted, and PI3K, pAkt and pmTOR protein expression level was suppressed in osteosarcoma cells by honokiol. The present study demonstrated, to the best of the authors' knowledge, for the first time that honokiol induces autophagy and apoptosis of osteosarcoma cells through the PI3K/Akt/mTOR signaling pathway.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Compostos de Bifenilo/farmacologia , Lignanas/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Neoplasias Ósseas/metabolismo , Caspase 3 , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ciclina D1/genética , Ciclina D1/metabolismo , Humanos , Osteossarcoma/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
OBJECTIVE: To investigate the morphological characteristics of the bridge tendon grafting in no man's land to reconstruct the tendon defect and the effect of passive mobilization on it. METHODS: A 2 cm defect was made in bilateral flexor digitorum profundus tendons of the middle chicken toes, and was then transplanted to the opposite site to serve as a segmental autograft tendon. Postoperatively, passive mobilization of the left and right middle toes began at 5 and 21 d separately. Specimens were studied by light, scanning and transmission electron microscopy at 5, 10, 21 and 35 d. RESULTS: Early repair of the tendon-graft of the left middle toes was made by proliferation and ingrowth of the epitenon cells intermingled with newly-formed collagen fibers. A gliding surface formed at 10 and 21 d. The tendon graft itself played an active role in the repair. In contrast, adhesions obliterated the surface and occupied the space between the tendon graft and surrounding tissues in the right middle toes. CONCLUSIONS: It indicates that the use of the segmental bridge tendon graft in no man's land coupled with early passive motion stimulates an intrisic repair process in both the tendon stump and the autogenous tendon graft and results in a functional healing.
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OBJECTIVE: To compare the immunological function and acute toxicity of the tissue cultured protocorm from Dendrobium candidum with natural medicinal materials from Dendrobium candidum. METHODS: The effect on immunological function was examined by counting white blood cells, weighing the weight of immune organs, and using carbon granules clearance and lymphocyte transformation test in mice treated with cyclophospamide. The acute toxicity was studied by giving maximum tolerated dose. RESULTS: The tissue cultured protocorm could increase the quantity of white blood cells, the ratios of thymus weight to body weight and spleen weight to body weight, promote the function of phagocytes and enhance the lymphocyte transformation rate. The mice could tolerate the dose of 54.56 g/kg(dried herbs) by oral administration. The functions were similar to those of natural medicinal materials from Dendrobium candidum. CONCLUSION: Both tissue cultured protocorm and natural medicinal materials from Dendrobium candidum could improve immunological function with similar potency. The maximum tolerated dose was 227 times as high as the effective clinical.
Assuntos
Adjuvantes Imunológicos/farmacologia , Dendrobium/química , Medicamentos de Ervas Chinesas/farmacologia , Plantas Medicinais/química , Adjuvantes Imunológicos/toxicidade , Animais , Técnicas de Cultura , Dendrobium/crescimento & desenvolvimento , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/toxicidade , Feminino , Contagem de Leucócitos , Ativação Linfocitária/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Camundongos , Fagocitose/efeitos dos fármacos , Caules de Planta/química , Caules de Planta/crescimento & desenvolvimento , Plantas Medicinais/crescimento & desenvolvimento , Distribuição AleatóriaRESUMO
OBJECTIVE: To investigate the effects and the related mechanisms of the components of Dang-Gui-Bu-Xue decoction (DGBXD) on improving blood deficiency. METHOD: The effects of promoting hematopoietic function were observed with the blood difficient model mice, by giving components of DGBXD. RBC, WBC, reticulocytes and bone marrow nucleated cells (BMNC) were determined. The components of DGBXD on proliferation of BMNC and on clony forming unit (CFU) were also determined. RESULT: The components of DGBXD remarkably increased the quantity of RBC, WBC, and BMNC. Some of the components promoted the proliferation of BMNC and increased the quantity of CFU-Mix. Among them, polysaccharide of angelica was most potent. CONCLUSION: The studies show that the extracts and some components of DGBXD can promote the hemopoietic function system of the model mice, and they exert the effects in a comprehensive way.
Assuntos
Angelica sinensis/química , Astragalus propinquus/química , Medicamentos de Ervas Chinesas/farmacologia , Células-Tronco Hematopoéticas/efeitos dos fármacos , Fitoterapia , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Animais , Contagem de Células Sanguíneas , Células da Medula Óssea/efeitos dos fármacos , Ensaio de Unidades Formadoras de Colônias , Combinação de Medicamentos , Feminino , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos ICR , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologiaRESUMO
Osteosarcoma is a high-grade malignant bone tumor. Pterostilbene (PTE) is a natural, dimethylated analog of resveratrol with higher bioavailability. While PTE has been shown to have potent antitumor activity against various types of cancer, the molecular mechanisms underlying the effects of PTE remain largely unknown. The Janus kinase 2/Signal Transducer and Activator of Transcription 3 (JAK2/STAT3) signaling pathway plays a crucial role in tumorigenesis and immune development. In this study, we assessed the antitumor activity of PTE against human osteosarcoma cells and explored the role of JAK2/STAT3 and apoptosis-related signaling pathways on the activity of PTE. PTE treatment resulted in a dose- and time-dependent inhibition of osteosarcoma cell viability. Additionally, PTE exhibited strong antitumor activity, as evidenced not only by reductions in tumor cell adhesion, migration and mitochondrial membrane potential (MMP) but also by increases in the apoptotic index, reactive oxygen species (ROS) and several biochemical parameters. Furthermore, PTE treatment directly inhibited the phosphorylation of JAK2 at Tyr 1007 and the downstream activation of STAT3. PTE also down-regulated the expression of STAT3 target genes, including the anti-apoptotic proteins Bcl-xL and Mcl-1, leading to the up-regulation of mitochondrial apoptosis pathway-related proteins (Bax, Bak, cytosolic Cytochrome c, and cleaved Caspase3) and cyclin-dependent kinase inhibitors such as p21 and p27. PTE, used in combination with a known JAK2/STAT3 inhibitor, AG490, further decreased the viability of osteosarcoma cells. Taken together, PTE is a potent inhibitor of osteosarcoma cell growth that targets the JAK2/STAT3 signaling pathway. These data suggest that inhibition of JAK2/STAT3 signaling is a novel mechanism of action for PTE during therapeutic intervention in osteosarcoma cancers.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Janus Quinase 2/antagonistas & inibidores , Osteossarcoma/tratamento farmacológico , Fator de Transcrição STAT3/antagonistas & inibidores , Estilbenos/farmacologia , Antineoplásicos Fitogênicos/administração & dosagem , Apoptose/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Osteossarcoma/patologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Estilbenos/administração & dosagem , Fatores de Tempo , Tirfostinas/farmacologia , Regulação para Cima/efeitos dos fármacosRESUMO
BACKGROUND: Cartilage degradation is a typical characteristic of arthritis. This study examined whether there was a subset of phagocytic chondrocytes that expressed the specific macrophage marker, CD163, and investigated their role in cartilage degradation. METHODS: Cartilage from the knee and temporomandibular joints of Sprague-Dawley rats was harvested. Cartilage degradation was experimentally-induced in rat temporomandibular joints, using published biomechanical dental methods. The expression levels of CD163 and inflammatory factors within cartilage, and the ability of CD163(+) chondrocytes to conduct phagocytosis were investigated. Cartilage from the knees of patients with osteoarthritis and normal cartilage from knee amputations was also investigated. RESULTS: In the experimentally-induced degrading cartilage from temporomandibular joints, phagocytes were capable of engulfing neighboring apoptotic and necrotic cells, and the levels of CD163, TNF-α and MMPs were all increased (P<0.05). However, the levels of ACP-1, NO and ROS, which relate to cellular digestion capability were unchanged (P>0.05). CD163(+) chondrocytes were found in the cartilage mid-zone of temporomandibular joints and knee from healthy, three-week old rats. Furthermore, an increased number of CD163(+) chondrocytes with enhanced phagocytic activity were present in Col-II(+) chondrocytes isolated from the degraded cartilage of temporomandibular joints in the eight-week experimental group compared with their age-matched controls. Increased number with enhanced phagocytic activity of CD163(+) chondrocytes were also found in isolated Col-II(+) chondrocytes stimulated with TNF-α (P<0.05). Mid-zone distribution of CD163(+) cells accompanied with increased expression of CD163 and TNF-α were further confirmed in the isolated Col-II(+) chondrocytes from the knee cartilage of human patients with osteoarthritis, in contrast to the controls (both P<0.05). CONCLUSIONS: An increased number of CD163(+) chondrocytes with enhanced phagocytic activity were discovered within degraded joint cartilage, indicating a role in eliminating degraded tissues. Targeting these cells provides a new strategy for the treatment of arthritis.
Assuntos
Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Cartilagem Articular/patologia , Condrócitos/metabolismo , Fagocitose , Receptores de Superfície Celular/metabolismo , Adulto , Idoso , Animais , Cartilagem Articular/metabolismo , Contagem de Células , Movimento Celular/efeitos dos fármacos , Separação Celular , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Feminino , Humanos , Articulação do Joelho/efeitos dos fármacos , Articulação do Joelho/metabolismo , Articulação do Joelho/patologia , Masculino , Pessoa de Meia-Idade , Osteoartrite/metabolismo , Osteoartrite/patologia , Fagocitose/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Articulação Temporomandibular/metabolismo , Articulação Temporomandibular/patologia , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
Hyperspectral reflectance (350-2500 nm) measurements were made over two experimental rice fields containing two cultivars treated with three levels of nitrogen application. Four different transformations of the reflectance data were analyzed for their capability to predict rice biophysical parameters, comprising leaf area index (LAI; m(2) green leaf area m(-2) soil) and green leaf chlorophyll density (GLCD; mg chlorophyll m(-2) soil), using stepwise multiple regression (SMR) models and support vector machines (SVMs). Four transformations of the rice canopy data were made, comprising reflectances (R), first-order derivative reflectances (D1), second-order derivative reflectances (D2), and logarithm transformation of reflectances (LOG). The polynomial kernel (POLY) of the SVM using R was the best model to predict rice LAI, with a root mean square error (RMSE) of 1.0496 LAI units. The analysis of variance kernel of SVM using LOG was the best model to predict rice GLCD, with an RMSE of 523.0741 mg m(-2). The SVM approach was not only superior to SMR models for predicting the rice biophysical parameters, but also provided a useful exploratory and predictive tool for analyzing different transformations of reflectance data.
Assuntos
Tomada de Decisões , Modelos Teóricos , Nitrogênio/análise , Oryza/química , Telemetria/métodos , Agricultura/métodos , Algoritmos , Biofísica , Clorofila/análise , Produtos Agrícolas , Oryza/anatomia & histologia , Folhas de Planta/anatomia & histologia , Folhas de Planta/química , Análise Espectral/métodosRESUMO
A carbon-carbon bond construction method is disclosed which involves radical addition of alpha-haloesters or iodoacetonitrile to enamides. Despite the presence of tri-n-butylstannane, nonreductive addition was predominant; H-atom transfer from tin hydride was not observed. Rapid iodine atom transfer to (or electron transfer from) the radical adduct, resulting in an iminium ion intermediate and radical chain propagation, is consistent with the observed reactivity.
RESUMO
OBJECTIVE: To explore the preventing effects of TGF-beta1 antibody (TGF-beta1Ab) compounded with fibrin glue (FG) on postoperative adhesions of flexor tendon. METHODS: Seventy-two Leghorn chickens were randomly divided into 4 groups (groups A, B, C and D), 18 chickens for each group, and the long flexor tendons of the 3rd and 4th toes in zone II of all chickens were transversed and sutured with the 4-strand cruciate repair technique to make defect models. In group A, 0.2 mL TGF-beta1 Ab was applied at repair site. In group B, 0.2 mL FG was applied at repair site. In group C, 0.2 mL TGF-beta1Ab and FG was applied at repair site. In group D, 0.2 mL normal sodium was applied at repair site. At 1, 3 and 8 weeks after operation, the tendons of 6 chickens in each group were harvested for morphological and histological evaluation. Six specimens of each group were obtained for biomechanical test at 3 and 8 weeks. RESULTS: The gross-observation showed that the differences in grading of tendon adhesion were not significant among 4 groups at 1 week after operation (P > 0.05), but the differences were significant between groups A, B, D and group C at 3 and 8 weeks after operation (P < 0.05). Histological observation showed that collagen fibers arranged irregularly in groups A, B and D, but arranged regularly in group C at 3 and 8 weeks' after operation. At 3 weeks after operation the gliding excursion ratio of the tendon in groups A, B, C and D were 0.45 +/- 0.05, 0.40 +/- 0.10, 0.79 +/- 0.09 and 0.25 +/- 0.07 respectively; the simulated active flexion ratio were 0.61 +/- 0.02, 0.67 +/- 0.03, 0.91 +/- 0.03 and 0.53 +/- 0.04 respectively; the work of flexion were (18.00 +/- 0.77), (17.80 +/- 1.13), (27.60 +/- 1.73) and (15.60 +/- 1.27) degrees/N respectively. There were significant differences between group C and other three groups (P < 0.05). The tendon anastomosis breaking strength were (14.2 +/- 1.9), (15.2 +/- 2.2), (16.0 +/- 2.2) and (14.7 +/- 2.7) N, showing no significant differences among 4 groups (P > 0.05). At 8 weeks after operation, the gliding excursion ratio of the tendon in groups A, B, C and D were 0.45 +/- 0.07, 0.43 +/- 0.08, 0.80 +/- 0.09 and 0.29 +/- 0.05 respectively; the simulated active flexion ratio were 0.61 +/- 0.02, 0.63 +/- 0.03, 0.92 +/- 0.03 and 0.53 +/- 0.03 respectively, the work of flexion were (18.30 +/- 0.84), (18.60 +/- 0.80), (27.90 +/- 1.24) and (15.30 +/- 0.75) degrees/N respectively. There were significant differences between group C and other three groups (P < 0.05). The tendon anastomosis breaking strength were(51.9 +/- 3.0), (51.4 +/- 1.4), (53.3 +/- 1.3) and (52.3 +/- 2.2) N, showing no significant differences among 4 groups (P > 0.05). CONCLUSION: TGF-beta1Ab compounded with FG could significantly prohibit the formation of fibrous adhesions without interfering with the healing process.