RESUMO
OBJECTIVE: To study the correlation of daily living activities with location and severity of trau- matic brain injury (TBI) and to provide a theoretical basis for improving the accuracy of expert opinion. METHODS: Five hundred and one cases of patients with TBI were selected. Detailed records included following: pre-injury situation, location and severity of injury, treatment and education. Daily living activi- ties scale (Barthel index) was applied to test the subjects' daily living activities. The relevance among location and severity of TBI and Barthel index was statistically analyzed. RESULTS: In mild TBI group, there was no significant difference in Barthel index among each location (P>0.05). In moderate TBI group, there were significant differences in Barthel index between subarachnoid hemorrhage and cerebral lobe injury, also between parietal, occipital lobes injury and frontal lobe injury, parietal, occipital lobes injury and temporal lobe (P<0.05), respectively, whereas no significant difference in Barthel index between frontal lobe injury and temporal lobe injury (P>0.05). In severe TBI, there were significant differences in Barthel index between every two different locations (P<0.05). CONCLUSION: There is some correlation between the location of TBI and Barthel index, which provides an important reference value for analyzing and determining daily living activities after TBI.
Assuntos
Atividades Cotidianas , Lesões Encefálicas/reabilitação , Avaliação de Resultados em Cuidados de Saúde , Adulto , Feminino , Humanos , Masculino , Índices de Gravidade do TraumaRESUMO
Freshwater planarian flatworm possesses an extraordinary ability to regenerate lost body parts after amputation; it is perfect organism model in regeneration and stem cell biology. Recently, small RNAs have been an increasing concern and studied in many aspects, including regeneration and stem cell biology, among others. In the current study, the large-scale cloning and sequencing of sRNAs from the intact and regenerative planarian Dugesia japonica are reported. Sequence analysis shows that sRNAs between 18nt and 40nt are mainly microRNAs and piRNAs. In addition, 209 conserved miRNAs and 12 novel miRNAs are identified. Especially, a better screening target method, negative-correlation relationship of miRNAs and mRNA, is adopted to improve target prediction accuracy. Similar to miRNAs, a diverse population of piRNAs and changes in the two samples are also listed. The present study is the first to report on the important role of sRNAs during planarian Dugesia japonica regeneration.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , Planárias/genética , RNA de Helmintos/genética , Pequeno RNA não Traduzido/genética , Animais , Sequência de Bases , Mapeamento Cromossômico , Perfilação da Expressão Gênica , Genoma Helmíntico/genética , MicroRNAs/genética , Dados de Sequência Molecular , Planárias/fisiologia , RNA Interferente Pequeno/genética , Regeneração/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Fibrolase is a non-hemorrhagic zinc metalloproteinase found in southern copperhead snake (Agkistrodon contortrix contortrix) venom that acts directly on fibrin clots and does not require plasminogen or any other blood-borne intermediate for activity. Chimeras of fibrolase with RGD peptides conferring antiplatelet activity have been synthesized covalently, but we describe a simpler, cheaper and less toxic method, using site-directed mutagensis. Fibrolase variants that constitute the arginine-glycine-aspartic acid (Arg-Gly-Asp, RGD) motif were constructed using site-directed mutagenesis. Chimeric genes of fibrolase were expressed in Escherichia coli to obtain the bifunctional chimeric molecule of fibrolase that can inhibit platelet aggregation. After refolding and purification, platelet-targeted thrombolysis and antiplatelet aggregation of the target chimeric protein were determined. The mutant RGD-F2, using the GPRGDWRMLG peptide to replace the TSVSHD sequence between sites 69 and 72, not only had almost the same catalytic ability as wild-type fibrolase but also a strong ability to inhibit platelet aggregation.
Assuntos
Agkistrodon/genética , Fibrinolíticos/farmacologia , Metaloendopeptidases/genética , Metaloendopeptidases/farmacologia , Oligopeptídeos/genética , Inibidores da Agregação Plaquetária/farmacologia , Sequência de Aminoácidos , Animais , Escherichia coli/genética , Fibrinólise/efeitos dos fármacos , Fibrinolíticos/metabolismo , Expressão Gênica , Humanos , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Agregação Plaquetária/efeitos dos fármacos , Inibidores da Agregação Plaquetária/metabolismo , Redobramento de Proteína , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Alinhamento de SequênciaRESUMO
Ancrod, a serine protease purified from the venom of Agkistrodon rhodostoma, is highly specific for fibrinogen. It causes anticoagulation by defibrinogenation and has been used as a therapeutic anticoagulant for the treatment of moderate to severe forms of peripheral arterial circulatory disorders in a variety of countries. The DNA of ancrod was amplified by recursive PCR with a yeast bias codon and cloned into the pGEM-T Easy vector. In order to achieve a high level secretion and a full activity expression of ancrod in Pichia pastoris (P. pastoris), the P. pastoris protein disulfide bond isomerase (PpPDI) was co-overexpressed in the strain. The secretion characteristics of ancrod with and without PpPDI were examined. With co-overexpression of PpPDI, the production of recombinant ancrod (rAncrod) was increased to 315 mg/L in the culture medium, which is twofold higher than the control strain carrying only the ancrod gene. Through purified by Ni²âº affinity chromatography and phenyl Sepharose column, the purity of rAncrod was found to be as high as 95.2%. The fibrinogenolytic and zymographic activities of the rAncrod were determined and found to be similar to that of the native protein. This improved expression system can facilitate further studies and the industrial production of ancrod.
Assuntos
Agkistrodon/metabolismo , Ancrod/metabolismo , Venenos de Crotalídeos/enzimologia , Proteínas Fúngicas/metabolismo , Pichia/enzimologia , Isomerases de Dissulfetos de Proteínas/metabolismo , Proteínas Recombinantes/metabolismo , Serina Endopeptidases/metabolismo , Ancrod/química , Ancrod/genética , Animais , Anticoagulantes/metabolismo , Western Blotting , Cromatografia de Afinidade , Venenos de Crotalídeos/química , Venenos de Crotalídeos/genética , Eletroforese em Gel de Poliacrilamida , Fibrinogênio/metabolismo , Proteínas Fúngicas/genética , Expressão Gênica , Vetores Genéticos , Pichia/genética , Reação em Cadeia da Polimerase , Isomerases de Dissulfetos de Proteínas/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Serina Endopeptidases/química , Serina Endopeptidases/genéticaRESUMO
Planarians exhibit an extraordinary ability to regenerate lost body parts which is attributed to an abundance of pluripotent somatic stem cells called neoblasts. In this article, we report a transcriptome sequence of a Planaria subspecies Dugesia japonica derived by high-throughput sequencing. In addition, we researched transcriptome changes during different periods of regeneration by using a tag-based digital gene expression (DGE) system. Consequently, 11,913,548 transcriptome sequencing reads were obtained. Finally, these reads were eventually assembled into 37,218 unique unigenes. These assembled unigenes were annotated with various methods. Transcriptome changes during planarian regeneration were investigated by using a tag-based DGE system. We obtained a sequencing depth of more than 3.5million tags per sample and identified a large number of differentially expressed genes at various stages of regeneration. The results provide a fairly comprehensive molecular biology background to the research on planarian development, particularly with regard to its regeneration progress.
