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1.
Adv Healthc Mater ; 10(12): e2100198, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33938637

RESUMO

Photodynamic therapy (PDT) often suffers from the exacerbated tumor hypoxia and the heterogeneous distribution of photosensitizers, leading to an inefficient ROS productivity and availability. In this work, a mitochondria targeted O2 economizer (designated as Mito-OxE) is developed to improve PDT efficiency by alleviating tumor hypoxia and enhancing the subcellular localization of photosensitizers. Specifically, the photosensitizer of protoporphyrin IX (PpIX) is modified with the hydrophilic polyethylene glycol and the lipophilic cation of triphenylphosphine (TPP) to fabricate the biocompatible mitochondria targeted photosensitizers (designated as Mito-PSs). And Mito-OxE is prepared by using Mito-PSs to load the mitochondrial oxidative phosphorylation inhibitors of atovaquone (ATO). Benefiting from the targeting capability of TPP, Mito-OxE can selectively accumulate in mitochondria after cellular uptake. Subsequently, the mitochondrial respiration would be suppressed to with the participation of ATO, resulting in a local hypoxia mitigation for enhanced PDT. Compared with Mito-PSs, Mito-OxE maximizes the therapeutic effect against hypoxic tumors under light irradiation. This design of mitochondria targeted O2 economizer would advance the development of targeted drug delivery system for effective PDT regardless of hypoxic microenvironment.


Assuntos
Nanopartículas , Fotoquimioterapia , Linhagem Celular Tumoral , Humanos , Hipóxia/metabolismo , Mitocôndrias/metabolismo , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Hipóxia Tumoral
2.
Nano Lett ; 20(3): 2062-2071, 2020 03 11.
Artigo em Inglês | MEDLINE | ID: mdl-32096643

RESUMO

Tumor hypoxia is the Achilles heel of oxygen-dependent photodynamic therapy (PDT), and tremendous challenges are confronted to reverse the tumor hypoxia. In this work, an oxidative phosphorylation inhibitor of atovaquone (ATO) and a photosensitizer of chlorine e6 (Ce6)-based self-delivery nanomedicine (designated as ACSN) were prepared via π-π stacking and hydrophobic interaction for O2-economized PDT against hypoxic tumors. Specifically, carrier-free ACSN exhibited an extremely high drug loading rate and avoided the excipient-induced systemic toxicity. Moreover, ACSN not only dramatically improved the solubility and stability of ATO and Ce6 but also enhanced the cellular internalization and intratumoral permeability. Abundant investigations confirmed that ACSN effectively suppressed the oxygen consumption to reverse the tumor hypoxia by inhibiting mitochondrial respiration. Benefiting from the synergistic mechanism, an enhanced PDT effect of ACSN was observed on the inhibition of tumor growth. This self-delivery system for oxygen-economized PDT might be a potential appealing clinical strategy for tumor eradication.


Assuntos
Neoplasias Mamárias Experimentais , Nanomedicina , Nanopartículas , Fotoquimioterapia , Porfirinas , Animais , Hipóxia Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Feminino , Neoplasias Mamárias Experimentais/tratamento farmacológico , Neoplasias Mamárias Experimentais/metabolismo , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Endogâmicos BALB C , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Nanopartículas/química , Nanopartículas/uso terapêutico , Porfirinas/química , Porfirinas/farmacocinética , Porfirinas/farmacologia
3.
Biomaterials ; 224: 119497, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31541935

RESUMO

In recent years, epigenetics has attracted great attentions in the field of biomedicine, which is used to denote the heritable changes in gene expression without any variation in DNA sequence, including DNA methylation, histone modification and so on. Inspired by it, a simple and versatile amino acids modification strategy is proposed in this paper to regulate the subcellular distribution of photosensitizer for plasma membrane targeted photodynamic therapy (PDT). Particularly, the plasma membrane anchoring ability and photo toxicity of the photosensitizer against different cell lines could be effectively manipulated at a single amino acid level. Systematic researches indicate that the number and variety of amino acids have a significant influence on the plasma membrane targeting effect of the photosensitizer. Furthermore, after self-assembling into nanoparticles, the obtained nano photosensitizers (NPs) also exhibit a good biocompatibility and plasma membrane targeting ability, which are conducive to enhancing the PDT therapeutic effect under light irradiation. Both in vitro and in vivo investigations confirm a robust tumor inhibition effect of NPs with a good biocompatibility. This epigenetics-inspired photosensitizer modification strategy would contribute to the development of structure-based drug design for tumor precision therapy.


Assuntos
Membrana Celular/metabolismo , Epigênese Genética , Neoplasias/tratamento farmacológico , Neoplasias/genética , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Células 3T3 , Aminoácidos/metabolismo , Animais , Linhagem Celular Tumoral , Membrana Celular/efeitos dos fármacos , Epigênese Genética/efeitos dos fármacos , Humanos , Camundongos , Nanopartículas/química , Nanopartículas/ultraestrutura , Fármacos Fotossensibilizantes/farmacologia , Protoporfirinas/farmacologia , Protoporfirinas/uso terapêutico , Esferoides Celulares/efeitos dos fármacos , Esferoides Celulares/patologia , Distribuição Tecidual/efeitos dos fármacos
4.
Biomaterials ; 211: 14-24, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31078049

RESUMO

Targeted delivery of the drug to its therapeutically active site with low immunogenicity and system toxicity is critical for optimal tumor therapy. In this paper, exosomes as naturally-derived nano-sized membrane vesicles are engineered by chimeric peptide for plasma membrane and nucleus targeted photosensitizer delivery and synergistic photodynamic therapy (PDT). Importantly, a dual-stage light strategy is adopted for precise PDT by selectively and sequentially destroying the plasma membrane and nucleus of tumor cells. Briefly, plasma membrane-targeted PDT of chimeric peptide engineered exosomes (ChiP-Exo) could directly disrupt the membrane integrity and cause cell death to some extent. More interestingly, the photochemical internalization (PCI) and lysosomal escape triggered by the first-stage light significantly improve the cytosolic delivery of ChiP-Exo, which could enhance its nuclear delivery due to the presence of nuclear localization signals (NLS) peptide. Upon the second-stage light irradiation, the intranuclear ChiP-Exo would activate reactive oxygen species (ROS) in situ to disrupt nuclei for robust and synergistic PDT. Based on exosomes, this dual-stage light guided subcellular dual-targeted PDT strategy exhibits a greatly enhanced therapeutic effect on the inhibition of tumor growth with minimized system toxicity, which also provides a new insight for the development of individualized biomedicine for precise tumor therapy.


Assuntos
Exossomos/transplante , Neoplasias/terapia , Peptídeos/uso terapêutico , Fármacos Fotossensibilizantes/uso terapêutico , Animais , Linhagem Celular Tumoral , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Membrana Celular/patologia , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Núcleo Celular/patologia , Sistemas de Liberação de Medicamentos , Feminino , Humanos , Camundongos Endogâmicos BALB C , Neoplasias/metabolismo , Neoplasias/patologia , Peptídeos/administração & dosagem , Fotoquimioterapia , Fármacos Fotossensibilizantes/administração & dosagem
5.
Chin J Traumatol ; 16(4): 249-50, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23910682

RESUMO

Pneumocephalus is the presence of air in the cranial vault. The common etiologies of pneumocephalus are brain trauma and cranial surgery. We report a case of a 26-year-old man with brain trauma who developed diffuse pneumocephalus after sneezing. CT scan was performed on arrival, and the image showed subarachnoid hemorrhage without pneumocephalus. On the seventh day after a big sneeze brain CT scan was re-performed, which showed pneumocephalus. After another ten days of treatment, the patient was discharged without any symptoms.


Assuntos
Lesões Encefálicas/complicações , Pneumocefalia/etiologia , Espirro , Adulto , Lesões Encefálicas/diagnóstico por imagem , Lesões Encefálicas/terapia , Humanos , Masculino , Pneumocefalia/diagnóstico por imagem , Pneumocefalia/terapia , Tomografia Computadorizada por Raios X
6.
Mol Biol Rep ; 38(6): 4193-6, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21110107

RESUMO

Lipopolysaccharide (LPS) from gram negative bacteria plays an important role in the pathophysiology of neurodegenerative diseases. Many evidences showed that LPS-induced neuroinflammation is related to upregulation of NF-kappaB. Here, we report that long-term treatment of lower dosage LPS mainly causes upregulation of Id2 protein. As an inhibitor of cell differentiation, Id2 plays an import role in adult olfactory neurogenesis. However, Id2 protein in brain acts as two edges in a sword, persist over-expression of Id2 in brain can induce neurodamages and may be related to neurodegeneration.


Assuntos
Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Proteína 2 Inibidora de Diferenciação/metabolismo , Lipopolissacarídeos/administração & dosagem , Lipopolissacarídeos/farmacologia , Animais , Benzimidazóis/metabolismo , Western Blotting , Encéfalo/patologia , Morte Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Proteína 2 Inibidora de Diferenciação/genética , Injeções Intraperitoneais , Camundongos , Camundongos Endogâmicos C57BL , Propídio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo
7.
Nan Fang Yi Ke Da Xue Xue Bao ; 29(4): 623-6, 2009 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-19403379

RESUMO

OBJECTIVE: To observe the effect of acupuncture on the expression of extracellular signal-regulated protein kinases 1/2 (ERK1/2) in the subcutaneous fascia of SD rats. METHODS: Eighteen SD rats were randomly divided into 6 groups (n=3) including 5 acupuncture groups and a control group. The rats in the 5 acupuncture groups received electro-acupuncture therapy in the regions of the inguinal groove, and at 0, 1, 6, 12, and 36 h after the last therapy, the superfacial fascia surrounding the acupuncture point (about 1.5 cm in diameter) were collected. The fascia tissues at the corresponding sites and at the acupoint Zusanli (ST36) were obtained from the control rats. The expression of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2) in the tissues were detected by Western blotting. RESULTS: ERK1/2 and p-ERK1/2 expressions were detected in the tissues harvested from both the acupoint and the non-acupoint in the control rats with similar expression intensities. In the rats of each acupuncture group, ERK1/2 expression was significantly increased on the acupuncture side in comparison with the control side. CONCLUSION: The normal loose connective tissue may participate in tissue proliferation and differentiation possibly via phosphorylation of ERK. Acupuncture can promote the signal transduction pathway of ERK, which can be a possible mechanism for the effect of acupuncture in modulating the physiopathological conditions.


Assuntos
Terapia por Acupuntura , Fáscia/enzimologia , Regulação Enzimológica da Expressão Gênica , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Pele , Pontos de Acupuntura , Animais , Western Blotting , Feminino , Ratos , Ratos Sprague-Dawley , Fatores de Tempo
8.
Cell Biochem Funct ; 27(4): 238-42, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19384903

RESUMO

The translocations of lipopolysaccharide (LPS) from the gut and its effects on bone healing are usually of clinical interest during bone fracture. As already widely studied, Cyclooxygenase-2 (COX-2) is a key enzyme for prostaglandin E2 (PGE(2)) production, which induces the nuclear factor kappa B (NFkappaB) activation and is beneficial to fracture healing. In order to know their roles in skeletal regeneration, mouse MC3T3-E1 osteoblasts were treated with NFkappaB inhibitor BAY 11-7082 and sc791 (a selective COX-2 inhibitor), in the presence of LPS. Interestingly, LPS could induce osteoblasts proliferation through increasing NFkappaB activation and translocation. This induction was not related to COX-2 expression, suggesting that LPS-induced NFkappaB activation is independent of COX-2. It is possible that low concentration of LPS can act as a stimulating factor of the NFkappaB pathway in nonstimulated cells such as osteoblasts. COX-2 is not necessary for the NFkappaB pathway during LPS-induced proliferation of osteoblasts since sc791 had no effects on this induction. These studies provide insight into a potential mechanism by which LPS can affect bone tissue repair in the initial phase of inflammation.


Assuntos
Proliferação de Células/efeitos dos fármacos , Ciclo-Oxigenase 2/metabolismo , Lipopolissacarídeos/farmacologia , NF-kappa B/metabolismo , Osteoblastos/efeitos dos fármacos , Animais , Linhagem Celular , Camundongos , NF-kappa B/antagonistas & inibidores , Nitrilas/farmacologia , Osteoblastos/metabolismo , Transdução de Sinais , Sulfonas/farmacologia
9.
Cell Biochem Funct ; 26(5): 598-602, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18508388

RESUMO

The mitochondrial flavoprotein apoptosis-inducing factor (AIF) has proved to be either the main mediator of apoptosis or an anti-apoptotic factor via its putative oxidoreductase and peroxide scavenging activities. We report here that 100 muM hydrogen peroxide (H2O2) induced the proliferation of C2C12 myoblasts and over-expression of AIF simultaneously in vitro. Immunofluorescence showed that the over-expression of AIF was located in the cytoplasm. The immunopositive AIF was detected in nuclei 27 days after denervation of skeletal muscle, but in the cytoplasm it was detected 27 days after fiber-damaged skeletal muscle. AIF may be a factor involved in skeletal muscle regeneration.


Assuntos
Fator de Indução de Apoptose/fisiologia , Proteínas Mitocondriais/fisiologia , Desenvolvimento Muscular/fisiologia , Músculo Esquelético/fisiologia , Regeneração/fisiologia , Animais , Fator de Indução de Apoptose/biossíntese , Fator de Indução de Apoptose/genética , Linhagem Celular , Peróxido de Hidrogênio/metabolismo , Masculino , Camundongos , Proteínas Mitocondriais/biossíntese , Proteínas Mitocondriais/genética , Ratos , Ratos Sprague-Dawley
10.
Nan Fang Yi Ke Da Xue Xue Bao ; 27(1): 1-4, 2007 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-17259132

RESUMO

OBJECTIVE: To observe the distribution of neuronal nitric oxide synthase (nNOS)-immunopositive neurons in rat corpus striatum and their ultrastructural features. METHODS: Brain tissue specimens were obtained from normal SD rats, in which nNOS-immunopositive neurons were visualized by ABC immunocytochemistry and observed under immunoelectron microscope with pre-embedding staining. RESULTS: Under light microscope, nNOS-immunopositive neurons appeared brown with distinct profiles of the cell body and processes. These neurons, mostly medium-sized and small cells, were located mainly in the lateral region of the corpus striatum. Only a few immunopositive neurons were detected in the medial region of the corpus striatum. Immunohistochemistry and transmission electron microscopy identified the nNOS-immunopositive neurons as interneurons possessing large nuclei with small amount of cytoplasma. The immunopositive granules were visualized as black plaques, and the larger ones distributed mainly in the cell bodies, some with monolayer membrane encapsulation. The small granules did not have the encapsulation, scattering in perinuclear regions and under the cell membrane, but not in the cell body. The immunopositive granules were also found in the axons and dendrites, but not in the vesicles of the synapses. In addition, many immunopositive terminals were found close to the blood vessels. CONCLUSIONS: nNOS-immunopositive neurons in rat corpus striatum are mainly medium-sized and small cells as is typical of the interneurons. The immunopositive granules locate in the cytoplasma, axons and dendrites, and larger granules have membrane coating while small ones do not, possibly in relation to their functions.


Assuntos
Corpo Estriado/enzimologia , Neurônios/enzimologia , Óxido Nítrico Sintase Tipo I/metabolismo , Animais , Corpo Estriado/ultraestrutura , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Transmissão , Neurônios/ultraestrutura , Ratos , Ratos Sprague-Dawley
11.
Nan Fang Yi Ke Da Xue Xue Bao ; 26(11): 1577-82, 2006 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-17121704

RESUMO

OBJECTIVE: To culture interleukin-1beta (IL-1beta)-activated Schwann cells (SCs) with human hair keratins (HHKs) for artificial nerve bridge construction. METHODS: SCs purified by primary culture with or without IL-1beta activation were cultured with HHKs decorated by extracellular matrix (ECM), and the artificial nerve bridge was implanted into the defect of rat sciatic nerve. The morphology of the SCs cultured with HHKs was monitored by inverted microscope, scanning electron microscope and evaluated by immunocytochemical staining, and the expression of nerve growth factor (NGF) in the sciatic nerve was observed by in situ hybridization. RESULTS: Activated SCs showed better ability to adhere to the HHKs and grew well. The HHKs component in the artificial nerve bridge underwent degradation in the sciatic nerve defect after 3 to 4 weeks, and IL-1beta activation resulted in enhanced NGF expression in the SCs. CONCLUSION: The constructed artificial nerve bridge by three-dimensional culture of IL-1beta-activiated SCs with HHKs decorated by ECM promotes the repair of sciatic nerve defects and accelerates sciatic nerve regeneration.


Assuntos
Interleucina-1beta/farmacologia , Queratinas/farmacologia , Células de Schwann/efeitos dos fármacos , Engenharia Tecidual/métodos , Animais , Animais Recém-Nascidos , Axônios/fisiologia , Técnicas de Cultura de Células , Movimento Celular/fisiologia , Células Cultivadas , Cabelo/química , Humanos , Microscopia Eletrônica de Varredura , Fator de Crescimento Neural/biossíntese , Regeneração Nervosa/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Células de Schwann/metabolismo , Células de Schwann/ultraestrutura , Nervo Isquiático/lesões , Nervo Isquiático/fisiopatologia , Nervo Isquiático/cirurgia
12.
DNA Seq ; 16(5): 335-9, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16323266

RESUMO

RNA deletions may be easier to detect and more extensive than DNA deletions. Two large deletion fragments (1120 and 7811 bp) of mitochondrial RNA were observed in rat L6 muscle cells. At the site of the 1120 bp deletion, the remaining RNA fragment was re-linked by a short additional section (GGTATGAAGCT). These kinds of deletions were accelerated by oxidative stress and were not observed in mitochondrial DNA.


Assuntos
Estresse Oxidativo , RNA/genética , Animais , Linhagem Celular , DNA Mitocondrial/genética , RNA Mitocondrial , Ratos , Deleção de Sequência
13.
Di Yi Jun Yi Da Xue Xue Bao ; 25(11): 1384-6, 2005 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-16305961

RESUMO

OBJECTIVE: To explore the relationship between the differentiation of L6 myoblasts and oxidative stress. METHODS: MTT assay was used to determine the viability of L6 myoblasts, from which the total RNA was extracted for amplification of the myogenin gene fragment by RT-PCR. H(2)O(2)-induced morphological changes of the cells were observed. RESULTS: The myoblasts treated with low concentration of reactive oxygen (50 micromol/L H(2)O(2)) for 1 h exhibited accelerated cell growth (P<0.05), and treatment with 50 and 150 micromol/L H(2)O(2) induced the gene expression of myogenin, a molecular marker for differentiation of myoblasts. Morphological study revealed myotube formation and accelerated differentiation of the myoblasts induced by H(2)O(2). CONCLUSION: The reactive oxygen may serve as the intracellular signal molecules to induce the growth and differentiation of the myoblasts.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Mioblastos/citologia , Estresse Oxidativo/fisiologia , Animais , Células Cultivadas , Ratos
14.
Di Yi Jun Yi Da Xue Xue Bao ; 25(9): 1128-31, 1144, 2005 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-16174579

RESUMO

OBJECTIVE: To culture Schwann cells (SCs) and human hair keratins (HHKs) for artificial nerve bridge construction. METHODS: SCs were purified by primary culture and labeled with BrdU, which were then cultured with HHKs decorated by ECM. The artificial nerve bridge was implanted into the defect of sciatic nerve, beneath the skin, and in the skeletal muscles of SD rat, respectively. The morphology of the SCs cultured with HHKs was monitored by inverted microscope and evaluated by immunocytochemical staining. Growth of BrdU-labeled SCs in vivo was observed by immunocytochemical staining on paraffin sections. RESULTS: In vitro cultured SCs were capable of adhering to HHKs and grew well four weeks after implantation. The HHK component in the artificial nerve bridge underwent degradation in the defect of the sciatic nerve, beneath the skin, and in the skeletal muscles of SD rat, and SC survival and proliferation were verified. CONCLUSION: SCs can survive in three-dimensional culture with HHKs for construction of artificial nerve bridge to repair nerve defects.


Assuntos
Movimento Celular/fisiologia , Cabelo/química , Queratinas/farmacologia , Regeneração Nervosa/fisiologia , Células de Schwann/citologia , Animais , Animais Recém-Nascidos , Axônios/fisiologia , Células Cultivadas , Humanos , Tecido Nervoso , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/citologia , Nervo Isquiático/lesões , Nervo Isquiático/cirurgia
15.
Di Yi Jun Yi Da Xue Xue Bao ; 25(1): 66-70, 2005 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-15684001

RESUMO

OBJECTIVE: To study the effect of FK506 in promoting apoptosis of peripheral blood-derived macrophages activated by homogenate of allogenic nerve tissues. METHODS: Homogenate of the allogenic nerve tissues was prepared using the sciatic nerve and injected in one-month-old SD rats, from which the macrophages activated by the homogenate were collected from the abdominal cavity and cultured in vitro. The cells were divided into 4 groups according to different concentrations of FK506 for treatment, namely 0 (group A, control group), 0.25 ng/ml (group B), 0.5 ng/ml (group C), and 1.0 ng/ml (group D). The cells of the 4 groups were inoculated into 96-well plate respectively for detecting the viability of the macrophages by MTT assay and for morphological evaluation of the cell apoptosis by transmission electron microscopy and fluorescence microscopy. RESULTS: The cells in groups B and C exhibited reduced viability and signs of apoptosis, and necrosis was observed in group D. Transmission electron microscopy and fluorescence microscopy identified early apoptotic changes and the presence of apoptotic body in the macrophages. The apoptotic rates of groups B and C were much higher than that in group A found by flow cytometry. CONCLUSION: FK506 can promote the apoptosis of macrophage activated by allogenic nerve homogenate and reduce macrophage-mediated immunological rejection of peripheral nerve allograft.


Assuntos
Apoptose/efeitos dos fármacos , Macrófagos Peritoneais/citologia , Tecido Nervoso/química , Tacrolimo/farmacologia , Extratos de Tecidos/farmacologia , Animais , Animais Recém-Nascidos , Células Cultivadas , Feminino , Masculino , Ratos , Ratos Sprague-Dawley
16.
Di Yi Jun Yi Da Xue Xue Bao ; 24(1): 27-31, 2004 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-14724089

RESUMO

OBJECTIVE: To conduct an in vitro study to assess the preliminary possibility of using formalin-fixed, instead of fresh, human bone tissues for allografting. METHODS: Fresh cadaveric bone tissues were fixed by formalin for more than 6 months and dissected into 5 mmx5 mmx5 mm pieces and 5 mmx5 mmx40 mm sticks, followed by chemical treatments to prepare the allograft bone materials. When alls treatments were completed, the bone grafts were centrifuged and their properties and cellular compatibility assessed in comparison with the currently used bone grafts clinically. RESULTS: The residual formaldehyde of the fixed allograft bone material was much below the controlled level and no significant differences were noted between the bone graft materials tested in regard to the chemical and mechanical properties and biocompatibility. CONCLUSION: This material we have prepared may meet the clinical demands for bone grafting, with good biocompatibility and less chance for infection by pathological agents.


Assuntos
Transplante Ósseo , Fixação de Tecidos , Animais , Ciclo Celular , Divisão Celular , Formaldeído , Humanos , Concentração de Íons de Hidrogênio , Microscopia Eletrônica de Varredura , Mioblastos/fisiologia , Ratos , Transplante Homólogo
17.
Di Yi Jun Yi Da Xue Xue Bao ; 23(12): 1283-5, 1289, 2003 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-14678891

RESUMO

OBJECTIVE: To construct a recombinant eukaryotic expression vector of rat brain-derived neurotrophic factor receptor trkB gene. METHODS: Using the total RNA extracted from rat brain tissue as the template, the trkB gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) with a pair of specific primers containing the restriction sites of EcoRI and BamHI. The amplified fragment of trkB gene was digested with EcoRI and BamHI, and then subcloned into cloning vector pMD18-T and then expression vector pEGFP-C2. The recombinant plasmid was identified by restriction endonuclease analysis and PCR. RESULTS: The amplified DNA fragment was about 1 461 bp in length, and enzyme digestion and PCR analysis showed that trkB gene had been successfully cloned into the vectors pMD18-T and pEGFP-C2. CONCLUSION: The trkB gene of rat has been successfully amplified and cloned into the eukaryotic expression vector pEGFP-C2.


Assuntos
Receptor trkB/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Animais , Clonagem Molecular , Plasmídeos , Ratos , Ratos Wistar
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