RESUMO
Brucellosis is a worldwide zoonotic disease caused by Brucella spp. Human infection often results from direct contact with tissues from infected animals or by consumption of undercooked meat and unpasteurised dairy products, causing serious economic losses and public health problems. The male genitourinary system is a common involved system in patients with brucellosis. Among them, unilateral orchitis and epididymitis are the most common. Although the clinical and imaging aspect of orchi-epididymitis caused by brucellosis have been widely described, the cellular and molecular mechanisms involved in the damage and the immune response in testis and epididymis have not been fully elucidated. In this review, we first summarised the clinical characteristics of Brucella epididymo-orchitis and the composition of testicular and epididymal immune system. Secondly, with regard to the mechanism of Brucella epididymoorchitis, we mainly discussed the process of Brucella invading testis and epididymis in temporal and spatial order, including i) Brucella evades innate immune recognition of testicular PRRsï¼ii) Brucella overcomes the immune storm triggered by the invasion of testis through bacterial lipoproteins and virulence factors, and changes the secretion mode of cytokines; iii) Brucella breaks through the blood-testis barrier with the help of macrophages, and inflammatory cytokines promote the oxidative stress of Sertoli cells, damaging the integrity of BTB; iv) Brucella inhibits apoptosis of testicular phagocytes. Finally, we revealed the structure and sequence of testis invaded by Brucella at the tissue level. This review will enable us to better understand the pathogenesis of orchi-epididymitis caused by brucellosis and shed light on the development of new treatment strategies for the treatment of brucellosis and the prevention of transition to chronic form. Facing the testicle with immunity privilege, Brucella is like Bruce Lee in the movie Game of Death, winning is survival while losing is death.HIGHLIGHTSWe summarized the clinical features and pathological changes of Brucellaepididymoorchitis.Our research reveals the pathogenesis of Brucella epididymoorchitis, which mainly includes the subversion of testicular immune privilege by Brucella and a series of destructive reactions derived from it.As a basic framework and valuable resource, this study can promote the exploration of the pathogenesis of Brucella and provide reference for determining new therapeutic targets for brucellosis in the future.
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Brucella , Brucelose , Epididimite , Orquite , Animais , Humanos , Masculino , ZoonosesRESUMO
Odontogenesis-associated phosphoprotein (ODAPH) is a recently discovered enamel matrix protein. Our previous study demonstrated that knockouting out Odaph in mice resulted in enamel hypomineralization. To further investigate the effect of Odaph on enamel mineralization, we constructed an Odaph overexpression mouse model, controlled by an amelogenin promoter. Our histological analysis of OdaphTg mice revealed that the enamel layer was thinner than in WT mice. An uneven, thinner enamel layer was confirmed using micro-computed tomography (uCT). It was subsequently found that the Tomes' processes lost their normal morphology, resulting in the loss of the enamel prism structure. These results indicate that Odaph overexpression in ameloblasts led to enamel dysplasia. In conjunction with this, Odaph overexpression hindered Amelx secretion, and may result in endoplasmic reticulum stress. Interestingly, uCT revealed that enamel had higher mineral density at the secretory stage; due to this, we did the histological staining for the mineralization-related proteins Alkaline phosphatase (ALPL) and Runt-related transcription factor 2 (RUNX2). It was observed that these proteins were up-regulated in OdaphTg mice versus WT mice, indicating that Odaph overexpression led to abnormal enamel mineralization. To confirm this, we transfected ameloblast-like cell line (ALC) with Odaph overexpression lentivirus in vitro and identified that both Alpl and Runx2 were strikingly upregulated in OE-mus-Odaph versus OE-NC cells. We concluded that the ectopic overexpression of Odaph in ameloblasts led to abnormal enamel mineralization. In summary, Odaph profoundly influences amelogenesis by participating in enamel mineralization.
Assuntos
Ameloblastos , Subunidade alfa 1 de Fator de Ligação ao Core , Animais , Camundongos , Ameloblastos/metabolismo , Amelogênese , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Fosfoproteínas , Microtomografia por Raio-X , Esmalte Dentário/metabolismo , Densidade Óssea , Calcificação FisiológicaRESUMO
BACKGROUND: Mutation in Odontogenesis-associated phosphoprotein (ODAPH) has been reported to cause recessive hypomineralized amelogenesis imperfecta (AI) in human. However, the exact role of ODAPH in amelogenesis is still unknown. RESULTS: ODAPH was identified as a novel constituent of the atypical basal lamina located at the interface between maturation ameloblasts and the enamel by dual immunofluorescence staining of ODAPH and LAMC2. Odaph knockout mice were generated to explore the function of ODAPH in amelogenesis. Odaph-/- mice teeth showed severely attrition and reduced enamel mineralization. Histological analysis showed from transition or early-maturation stage, ameloblasts were rapidly shortened, lost cell polarity, and exhibited cell pathology. Abundant enamel matrix marked by amelogenin was retained. Temporary cyst-like structures were formed between flattened epithelial cells and the enamel from maturation stage to eruption. The integrity of the atypical basal lamina was impaired indicated by the reduced diffuse expression of LAMC2 and AMTN. The expression of maturation stage related genes of Amtn, Klk4, Integrinß6 and Slc24a4 were significantly decreased. CONCLUSIONS: Our results suggested Odaph played vital roles during amelogenesis by maintaining the integrity of the atypical basal lamina in maturation stage, which may contribute to a better understanding of the pathophysiology of human AI.
Assuntos
Amelogênese/genética , Esmalte Dentário/metabolismo , Proteínas da Matriz Extracelular/genética , Fosfoproteínas/genética , Ameloblastos/metabolismo , Animais , Proteínas da Matriz Extracelular/metabolismo , Laminina/genética , Laminina/metabolismo , Camundongos , Camundongos Knockout , Fosfoproteínas/metabolismoRESUMO
OBJECTIVE: The aim of this investigation was to study the effects of Runt-related transcription factor 2 (Runx2) on the junctional epithelium and alveolar bone. METHODS: The attachment level of the junctional epithelium and the resorption of alveolar bone were analyzed by histology and scanning electron microscopy. The expression of amelotin was determined by immunohistochemistry, Western blot, and real-time PCR. The ultrastructure of the dentogingival interface was observed by transmission electron microscopy. RESULTS: The cKO mice demonstrated remarkable attachment loss, epithelial hyperplasia, and alveolar bone loss. The relative protein and mRNA expression of amelotin was increased in the junctional epithelium of the cKO mice. The attachment apparatus of the cKO mice showed ultrastructural deficiency. CONCLUSIONS: Loss of Runx2 led to the junctional epithelium and alveolar bone defects in mice. Runx2 may play a crucial role in maintaining the integrity of the dentogingival junction and the normal structure of alveolar bone.
Assuntos
Perda do Osso Alveolar , Inserção Epitelial , Perda do Osso Alveolar/genética , Animais , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Epitélio , Camundongos , Camundongos KnockoutRESUMO
BACKGROUND: Hepatic fibrosis is a worldwide incurable disease; due to the complex and unclear mechanism, there lack the effective therapeutic targets. However, the mechanism of miR-23a-5p underling this pathological process is largely not clear. The purpose of this study was to investigate the role of miR-23a-5p in hepatic fibrosis and HSC activation. METHODS: The content of miR-23a-5p in hepatic fibrosis induced by N-nitrosodimethylamine (NDMA) and HSC activation induced by platelet-derived growth factor (PDGF) was detected by qRT-PCR. H&E staining, Masson staining and Shear wave electrography (SWE) were used to detect the degree of hepatic fibrosis. Immunohistochemistry staining, qRT-PCR and Western blot detect the related markers of liver fibrosis or HSC activation, as well as the related pathway genes and proteins. Dual-luciferase reporter system verifies the interaction between miR-23a-5p with PTEN or miR-23a-5p with lncRNA LOC102551149 in HSC-T6. siRNA and miRNA mimic transfer to HSC-T6 to detect the function of lncRNA LOC102551149 and miR-23a-5p on HSC activation. RESULTS: After hepatic fibrosis and HSC activation happened, the expression of miR-23a-5p was up-regulated, whereas anti-miR-23a-5p can alleviate hepatic fibrosis and HSC activation. Further research shows miR-23a-5p can target PTEN and degrade it, causing activation of PI3K/Akt/mTOR/Snail pathway. lncRNA LOC102551149 can be used as a competition endogenous RNA (ceRNA) targeting miR-23a-5p through base pairing, and siRNA LOC102551149 or exogenous miR-23a-5p can induce HSC activation through PI3K/Akt/mTOR/Snail pathway. CONCLUSION: We demonstrate mechanism pathway of miR-23a-5p on hepatic fibrosis and HSC activation, which may develop a therapeutic target for hepatic fibrosis.
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Células Estreladas do Fígado/metabolismo , Cirrose Hepática/genética , MicroRNAs/genética , RNA Longo não Codificante/metabolismo , Animais , Western Blotting , Linhagem Celular , Dimetilnitrosamina/toxicidade , Técnicas de Silenciamento de Genes , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , MicroRNAs/antagonistas & inibidores , MicroRNAs/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Hepatic fibrosis is chronic liver damage with many causes that has a relatively high death rate. The current study showed that long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5), microRNA-23a (miR-23a), and phosphatase and tensin homolog (PTEN) play important roles in the pathological process of hepatic fibrosis but have a relatively unclear regulatory mechanism. This study aimed to investigate the roles of lncRNA GAS5, miR-23a, and PTEN in the pathological process of hepatic fibrosis and hepatic stellate cell (HSC) activation. We used carbon tetrachloride (CCl4) intraperitoneal injections to establish a rat hepatic fibrosis model and exogenous transforming growth factor-ß1 to establish an HSC activation model. Quantitative RT-PCR, Western blot, dual-luciferase reporter system, and RNA pull-down assays were used to investigate which microRNAs and lncRNAs participate in the process of hepatic fibrosis and HSC activation. miR-23a expression increased significantly in hepatic fibrosis tissues and activated HSCs. miR-23a interaction with and degradation of PTEN further influenced the downstream signaling pathway phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin/Snail (PI3K/Akt/mTOR/Snail), causing E-cadherin expression levels to decrease and α-smooth muscle actin and collagen I expression levels to increase. lncRNA GAS5 can be used as a sponge platform for miR-23a to decrease miR-23a expression levels competitively. We revealed the role of the lncRNA GAS5/miR-23a/PTEN/PI3K/Akt/mTOR/Snail signaling pathway in hepatic fibrosis, providing molecular targets for the treatment of hepatic fibrosis. NEW & NOTEWORTHY This is the first study revealing that microRNA-23a (miR-23a) promotes hepatic fibrosis through the phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin/Snail signaling pathway, and long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) can act as a sponge platform for miR-23a. Therefore, lncRNA GAS5/miR-23a may bring molecular targets for hepatic fibrosis therapy.
Assuntos
Proteínas de Transporte/metabolismo , Células Estreladas do Fígado/metabolismo , Cirrose Hepática/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Nucleolar Pequeno/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Animais , Descoberta de Drogas , Células Estreladas do Fígado/efeitos dos fármacos , MicroRNAs/genética , RNA Longo não Codificante/genética , Ratos , Transdução de Sinais/efeitos dos fármacos , Fator de Crescimento Transformador beta/farmacologiaRESUMO
This study aimed to investigate the effect of oestrogen on prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) secretion in bovine oviduct epithelial cells. Bovine oviduct epithelial cells were obtained from the lumen of fresh bovine oviducts. Quantitative real-time polymerase chain reaction and in-cell western assays were used to measure PGE2 and PGF2α synthase activity and enzyme-linked immunosorbent assays were used to detect the concentrations of the two prostaglandins in extracellular fluid. We observed that oestradiol caused a short-term increase in cyclo-oxygenase-2 (COX-2), which stimulated PGE2 and PGF2α secretion, and that a subsequent decrease in COX-2 and an increase in cyclo-oxygenase-1 (COX-1) produced a high PGE2:PGF2α ratio. These findings reflect the dynamic change in PGE2 and PGF2α levels under the influence of oestrogen, which may be essential for fertilisation.
Assuntos
Dinoprosta/metabolismo , Dinoprostona/metabolismo , Células Epiteliais/efeitos dos fármacos , Estradiol/farmacologia , Oviductos/efeitos dos fármacos , Animais , Bovinos , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Células Epiteliais/metabolismo , Feminino , Hidroxiprostaglandina Desidrogenases/genética , Hidroxiprostaglandina Desidrogenases/metabolismo , Oviductos/metabolismoRESUMO
Gamete and embryo transport is an important function of the oviduct. This transport involves both smooth muscle contraction and epithelial cell secretions, the former of which is mediated by prostaglandins (PGs) and their receptors. Our aim was to study the regulation of prostaglandin E2 and prostaglandin F2α receptors (EP2, EP4, and FP receptor) by estradiol in bovine oviduct smooth muscle. EP2, EP4, and FP receptor mRNA and protein expression was investigated using real-time RT-PCR and Western blot analyses, respectively. To evaluate the contraction or relaxation of cultured bovine oviductal smooth muscle tissue, peristalsis was used to assess contractile activity. EP2, EP4, and FP receptor mRNA and protein expression was increased in oviductal smooth muscle tissue after treatment with different concentrations of estradiol for various durations. The expression of all receptors peaked at an estradiol concentration of 10(-11)mol/L after 8h of treatment, whereas no increase in expression was observed after fulvestrant (a selective antagonist of E2 receptor) treatment, indicating that E2 interacts with specific E2 nuclear receptors to regulate EP2, EP4, and FP receptor expression. Although PGF2α and PGE2 induced both contraction and relaxation, no significant differences were found in contractility between the estradiol-treated and control groups, with both groups of cultured smooth muscle strips showing similar vitality. In conclusion, estradiol increases EP2, EP4, and FP receptor mRNA and protein expression in bovine oviductal smooth muscle when added for different periods of time and at different concentrations. Additionally, E2 is transported intracellularly and interacts with specific E2 nuclear receptors to regulate their expression.
Assuntos
Estradiol/metabolismo , Músculo Liso/metabolismo , Oviductos/metabolismo , Receptores de Prostaglandina E Subtipo EP2/agonistas , Receptores de Prostaglandina E Subtipo EP4/agonistas , Receptores de Prostaglandina/agonistas , Regulação para Cima , Matadouros , Absorção Fisiológica/efeitos dos fármacos , Animais , Transporte Biológico/efeitos dos fármacos , Bovinos , China , Cruzamentos Genéticos , Estradiol/análogos & derivados , Estradiol/química , Estradiol/farmacologia , Antagonistas do Receptor de Estrogênio/farmacologia , Feminino , Fulvestranto , Contração Muscular/efeitos dos fármacos , Relaxamento Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Oviductos/efeitos dos fármacos , RNA Mensageiro/metabolismo , Receptores de Estradiol/agonistas , Receptores de Estradiol/antagonistas & inibidores , Receptores de Estradiol/metabolismo , Receptores de Prostaglandina/genética , Receptores de Prostaglandina/metabolismo , Receptores de Prostaglandina E Subtipo EP2/genética , Receptores de Prostaglandina E Subtipo EP2/metabolismo , Receptores de Prostaglandina E Subtipo EP4/genética , Receptores de Prostaglandina E Subtipo EP4/metabolismo , Técnicas de Cultura de Tecidos , Regulação para Cima/efeitos dos fármacosRESUMO
Background: With the advancement of society, the cultivation of medical professionals equipped with solid theoretical knowledge, a strong sense of innovation, and critical thinking has become a crucial goal in the reform of medical higher education. Over recent years, the hybrid Problem-Based Learning (hPBL) model, a blend of Problem-Based Learning (PBL) and Lecture-Based Learning (LBL), has emerged as a novel approach in the medical education reform landscape of China. The application and efficacy of the hPBL model in medical experimental courses have piqued the interest of medical educators. The aim of this study was to appraise the application and effectiveness of the hPBL model in the experimental course of Medical Molecular Biology at Beihua University. Methods: Utilizing the "Rain Classroom" platform, students from the Preventive Medicine and Medical Imaging programs were allocated to either the hPBL or LBL method for their Medical Molecular Biology experimental courses. The hPBL model's impact on students' performance was evaluated across four domains: experimental theory, experimental operation, experimental report, and practical application. Questionnaires were employed to gauge students' experiences and perceptions. Results: The results indicated that the final assessment scores of the hPBL group were significantly superior to those of the LBL group. Moreover, the hPBL model effectively amplified students' self-learning capability, practical application skills, and communication competencies. Students expressed a high degree of satisfaction with this blended learning model. Conclusion: The hPBL model, which amalgamates PBL and LBL, has demonstrated its effectiveness in medical education. Its implementation in the experimental course of Medical Molecular Biology at Beihua University yielded positive outcomes, enhancing students' performance and satisfaction levels. Consequently, it is recommended that the hPBL model be further promulgated in other medical experimental courses.
RESUMO
OBJECTIVE: Long-term and high exposure to UV radiation can lead to the development of skin photoaging diseases. Therefore, there is an ongoing need for more natural and safe drugs to prevent or treat skin photoaging diseases. METHODS: The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database were used to collect the active compounds and corresponding targets of Cnidii Fructus, Arnebiae Radix, Angelicae Sinensis Radix, Poria, and Borneolum. The GeneCards database and the NCBI Gene database were used to collect the targets of skin photoaging diseases. The STRING database was used to construct a protein-protein interaction network formed by the intersecting targets of drugs and diseases. The Metascape database was applied for Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis of the targets. Molecular docking between active compounds and targets was verified by Autodock. After that, the skin photoaging model of mice was established and treated with MP gel. The skin characterization on the back of mice was observed, and the ameliorative effect of MP gel on skin photoaging was evaluated by histological and epidermal thickness assays. The MDA content and SOD activity were measured. Caspase-3 expression in mouse skin tissues was detected by immunohistochemistry, quantitative real-time polymerase chain reaction assay, and Western blot. RESULTS: The results of network pharmacology experiments showed that the natural drugs have multi-component, multi-target therapeutic disease characteristics. The results of animal studies showed that MP gel improved the health of photoaged skin, promoted skin structural integrity, had antioxidant properties and significantly inhibited caspase-3 expression. CONCLUSION: The experimental validation of the results of the preliminary network pharmacology analysis was carried out in animal experiments, which confirmed part of the mechanism of action of MP gel in the prevention and treatment of skin photoaging.
Assuntos
Envelhecimento da Pele , Animais , Camundongos , Simulação de Acoplamento Molecular , Caspase 3 , Farmacologia em Rede , PeleRESUMO
Human infections caused by Brucella (called brucellosis) are among the most common zoonoses worldwide with an estimated 500,000 cases each year. Since chronic Brucella infections are extremely difficult to treat, there is an urgent need for more effective therapeutics. As a facultative intracellular bacterium, Brucella is strictly parasitic in the host cell. Here, we performed proteomic and transcriptomic and metabolomic analyses on Brucella infected patients, mice and cells that provided an extensive "map" of physiological changes in brucellosis patients and characterized the metabolic pathways essential to the response to infection, as well as the associated cellular response and molecular mechanisms. This is the first report utilizing multi-omics analysis to investigate the global response of proteins and metabolites associated with Brucella infection, and the data can provide a comprehensive insight to understand the mechanism of Brucella infection. We demonstrated that Brucella increased nucleotide synthesis in the host, consistent with increased biomass requirement. We also identified IMPDH2, a key regulatory complex that controls nucleotide synthesis during Brucella infection. Pharmacological targeting of IMPDH2, the rate-limiting enzyme in guanine nucleotide biosynthesis, efficiently inhibits B. abortus growth both in vitro and in vivo. Through screening a library of natural products, we identified oxymatrine, an alkaloid obtained primarily from Sophora roots, is a novel and selective IMPDH2 inhibitor. In further in vitro bacterial inhibition assays, oxymatrine effectively inhibited the growth of B. abortus, which was impaired by exogenous supplementation of guanosine, a salvage pathway of purine nucleotides. This moderately potent, structurally novel compound may provide clues for further design and development of efficient IMPDH2 inhibitors and also demonstrates the potential of natural compounds from plants against Brucella.
Assuntos
Brucella abortus , Brucelose , Humanos , Animais , Camundongos , Brucella abortus/metabolismo , Proteômica , Multiômica , Brucelose/microbiologia , Brucelose/prevenção & controle , Nucleotídeos/metabolismoRESUMO
Transforming growth factor ß1 (TGF-ß1) and Runt-related transcription factor 2 (RUNX2) are critical factors promoting enamel development and maturation. Our previous studies reported that absence of TGF-ß1 or RUNX2 resulted in abnormal secretion and absorption of enamel matrix proteins. However, the mechanism remained enigmatic. In this study, TGF-ß1-/-Runx2-/- and TGF-ß1+/-Runx2+/- mice were successfully generated to clarify the relationship between TGF-ß1 and RUNX2 during amelogenesis. Lower mineralization was observed in TGF-ß1-/-Runx2-/- and TGF-ß1+/-Runx2+/- mice than single gene deficient mice. Micro-computed tomography (µCT) revealed a lower ratio of enamel to dentin density in TGF-ß1-/-Runx2-/- mice. Although µCT elucidated a relatively constant enamel thickness, variation was identified by scanning electron microscopy, which revealed that TGF-ß1-/-Runx2-/- mice were more vulnerable to acid etching with lower degree of enamel mineralization. Furthermore, the double gene knock-out mice exhibited more serious enamel dysplasia than the single gene deficient mice. Hematoxylin-eosin staining revealed abnormalities in ameloblast morphology and arrangement in TGF-ß1-/-Runx2-/- mice, which was accompanied by the absence of atypical basal lamina (BL) and the ectopic of enamel matrix. Odontogenesis-associated phosphoprotein (ODAPH) has been identified as a component of an atypical BL. The protein and mRNA expression of ODAPH were down-regulated. In summary, TGF-ß1 and RUNX2 might synergistically regulate enamel mineralization through the downstream target gene Odaph. However, the specific mechanism by which TGF-ß1 and RUNX2 promote mineralization remains to be further studied.
Assuntos
Amelogênese , Fator de Crescimento Transformador beta1/metabolismo , Ameloblastos/metabolismo , Amelogênese/genética , Animais , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Camundongos , Odontogênese/fisiologia , Fosfoproteínas/metabolismo , Microtomografia por Raio-XRESUMO
Junctional epithelium (JE) attaching to the enamel surface seals gaps around the teeth, functioning as the first line of gingival defense. Runt-related transcription factor 2 (Runx2) plays a role in epithelial cell fate, and the deficiency of Runx2 in JE causes periodontal destruction, while its effect on the barrier function of JE remains largely unexplored. In the present study, hematoxylin-eosin (H&E) staining revealed the morphological differences of JE between wild-type (WT) and Runx2 conditional knockout (cKO) mice. We speculated that these changes were related to the down-regulation of E-cadherin (E-cad), junctional adhesion molecule 1 (JAM1), and integrin ß6 (ITGB6) in JE. Moreover, immunohistochemistry (IHC) was conducted to assess the expressions of these proteins. To verify the relationship between Runx2 and the three above-mentioned proteins, human gingival epithelial cells (HGEs) were cultured for in vitro experiment. The expression of Runx2 in HEGs was depleted by lentivirus. Quantitative real-time PCR (qRT-PCR) and Western blotting analysis were adopted to analyze the differences in mRNA and protein expressions. Taken together, Runx2 played a crucial role in maintaining the structure and function integrality of JE via regulating the expressions of E-cad and JAM1.
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Caderinas/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/deficiência , Epitélio/metabolismo , Moléculas de Adesão Juncional/metabolismo , Dente Molar/metabolismo , Animais , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Regulação para Baixo , Células Epiteliais/metabolismo , Gengiva/citologia , Humanos , Cadeias beta de Integrinas/metabolismo , Mandíbula/metabolismo , Camundongos Knockout , Periodonto/metabolismoRESUMO
OBJECTIVE: To establish a convenient and accurate method of DNA molecular marker for the identification of corium stomachium galli. METHOD: Cytb mtDNA sequences of Gallus gallus domestica and three other species of poultry were downloaded from Genbank. Species-specific PCR primers were designed according to the differential DNA fragments of the cytb genes. PCR tests were performed with the DNAs extracted from G. gallus domestica, three other poultry species and domestic mammal animals. RESULT: The specific primers of G. gallus domestica could only amplify the cytb mtDNA of G. gallus domestica. CONCLUSION: The primers are specific to G. gallus domestica mtDNA and can used to discern Corium stomachium from the false medicine.
Assuntos
Galinhas/classificação , Galinhas/genética , Animais , DNA Mitocondrial/genética , Fígado/metabolismo , Reação em Cadeia da PolimeraseRESUMO
Runt-related transcription factor 2 (Runx2) is involved in the early stage of tooth development. However, only few studies have reported the role of Runx2 in enamel development, which may be attributed to that Runx2 full knockout mice cannot survive after birth. In the present study, we successfully established a Runx2-deficient mouse model using a conditional knockout (cKO) method. We observed a significant reduction in the degree of mineralization and the decreased size of enamel rods in cKO mice. Histological analysis showed the retained enamel proteins in enamel layer at maturation stage in cKO molars. Further analysis by qRT-PCR revealed that the expressions of genes encoding enamel structure proteins, such as amelogenin (AMELX), ameloblastin (AMBN) and enamelin (ENAM), were increased in cKO enamel organs. On the other hand, the expression of kallikrein-related peptidase-4 (KLK4) at the mRNA and protein levels was dramatically decreased from late secretory stage to maturation stage in cKO enamel organs, while the expression of matrix metalloproteinase-20 (MMP-20) was not significantly altered. Finally, immunohistochemistry indicated that the uptake of amelogenins by ameloblasts was significantly decreased in cKO mice. Taken together, Runx2 played critical roles in controlling enamel maturation by increasing synthesis of KLK4 and decreasing synthesis of AMELX, AMBN and ENAM.
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Ameloblastos/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/deficiência , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Esmalte Dentário/citologia , Esmalte Dentário/crescimento & desenvolvimento , Técnicas de Inativação de Genes , Amelogenina/metabolismo , Animais , Esmalte Dentário/metabolismo , Proteínas do Esmalte Dentário/metabolismo , Regulação da Expressão Gênica , Calicreínas/metabolismo , Camundongos , Minerais/metabolismoAssuntos
Dente Impactado , Pé , Mãos , Humanos , Extremidade Inferior , Dente Impactado/cirurgia , Extremidade SuperiorRESUMO
PURPOSE: To study the feasibility of tissue engineered bone constructed with simvastatin carried by PLGA/CPC and bone marrow stromal cells (BMSCs) and screen the effective drug loading of simvastatin. METHODS: Solvent casting-particle leaching technology combined with the phase separation process was used to prepare the different concentrations (simvastatin mass: 0.1, 0.5, 1 mg) of simvastatin carried by PLGA/CPC composite scaffold materials. Scanning electron microscopy was used to observe the porosity and drug release curve was drawn; Alizarin red staining and type I collagen staining were applied to observe the effect of osteogenic medium and simvastatin on the role of BMSCs to the osteogenetic differentiation. The induced passage 3 cells after dil staining were mixed with the composite scaffold material to a complex. Scanning electron microscopy and laser confocal microscope were used to observe the adhesion on the complex. CCK-8 and alkaline phosphatase (ALP) were applied to observe the proliferation and differentiation. SPSS 18.0 software package was used for statistical analysis. RESULTS: The scaffold porosity was more than 90% with an average aperture of 200-300 µm. The drug released slowly. There was no obvious interpretation. Type I collagen showed positive expression. Alizarin red staining proofed the formation of mineralization nodules in group which was induced with the conditional medium and simvastatin. 0.5 mg group showed cells adhered to the inner surface of the scaffold material and could significantly promote the proliferation and differentiation of cells. CONCLUSIONS: Combination of simvastatin and osteogenic medium can effectively promote the differentiation of BMSCs. Simvastatin carried by PLGA/CPC scaffold materials is an ideal tissue engineering scaffold material. PLGA/CPC scaffold containing 0.5 mg simvastatin can effectively promote the proliferation and differentiation of BMSCs. Supported by Natural Science Foundation of Jilin Province (201215052).