RESUMO
Our study aims to explore the effects of lentivirus-mediated microRNA-124 (miR-124) gene-modified bone marrow mesenchymal stem cell (BMSC) transplantation on the repair of spinal cord injury (SCI) in rats. BMSCs were isolated from the bone marrow of rats. The target gene miR-124 was identified using a luciferase-reporter gene assay. Seventy-two rats were selected for construction of the SCI model, and the rats were randomly divided into the blank group, sham group, SCI group, negative control (NC) group, overexpressed miR-124 group and si-PDXK group. The mRNA expression of miR-124 and the mRNA and protein expression of pyridoxal kinase (PDXK) were detected by quantitative real-time polymerase chain reaction and western blotting. The locomotor capacity of the rats was evaluated using the Basso, Beattie and Bresnahan (BBB) scale. Brdu, neuron-specific enolase (NSE), neurofilament (NF) and microtubule-associated protein 2 (MAP2) were detected using immunohistochemistry. The expression levels of thyrotropin-releasing hormone (TRH), prostacyclin (PGI2) and gangliosides (GM) were measured using an enzyme-linked immunosorbent assay. PDXK was identified as the target gene of miR-124. The overexpressed miR-124 group exhibited higher miR-124 expression than the SCI, NC and si-PDXK groups. Compared with the SCI and NC groups, the PDXK expression was downregulated in the overexpressed miR-124 and si-PDXK groups, and the BBB scores were significantly increased 7, 21 and 35 days after transplantation. The double-labeled positive cell densities (Brdu+NSE/NF/MAP2) and the expression levels of TRH, PGI2 and GM in the overexpressed miR-124 group were significantly higher than those in the NC and SCI groups. These results indicated that miR-124 targeted PDXK to accelerate the differentiation of BMSCs into neurocytes and promote SCI repair.
Assuntos
Animais , Ratos , Western Blotting , Medula Óssea , Bromodesoxiuridina , Contagem de Células , Ensaio de Imunoadsorção Enzimática , Epoprostenol , Gangliosídeos , Imuno-Histoquímica , Filamentos Intermediários , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Proteínas Associadas aos Microtúbulos , Fosfopiruvato Hidratase , Piridoxal Quinase , Reação em Cadeia da Polimerase em Tempo Real , RNA Mensageiro , Traumatismos da Medula Espinal , Medula Espinal , Hormônio Liberador de TireotropinaRESUMO
<p><b>AIM</b>To identify the genes specifically expressed in human adult and fetal testes and spermatozoa.</p><p><b>METHODS</b>A human testis cDNA microarray was established. Then mRNAs of human adult and fetal testis and spermatozoa were purified and probes were prepared by a reverse transcription reaction with mRNA as the template. The microarray was hybridized with probes of adult and fetal testes and spermatozoa. The nucleic acid sequences of differentially expressed genes were determined and homologies were searched in the databases of GenBank.</p><p><b>RESULTS</b>A novel human testis-specific gene, PKH-T, was identified by hybridizing adult and fetal testis and spermatozoa probes with a human testis cDNA microarray. The cDNA of PKH-T was 1 069 bp in length. The cDNA sequence of this clone was deposited in the Genbank (AY303972) and PKH-T was also determined as Interim GenSymbol (Unigene, HS.38041). PKH-T contained most PKH conserved motif. The 239 amino acid sequences deduced from the 719 bp open reading frame (ORF) had a homology with the gene PKH (U89606). PKH-T was specifically and strongly expressed in the testis. Comparison of the differential expressions of PKH and PKH-T in testes of different developmental stages indicated that PKH-T was expressed in the adult testis and spermatozoa, while PKH, in the adult, fetal and aged testes. PKH-T had no expression in the testis of Sertoli cell only and partially spermatogenic arrest patients.</p><p><b>CONCLUSION</b>PKH-T is a gene highly expressed in adult human testis and spermatozoa. It may play an important role in spermatogenesis and could be related to male infertility.</p>
Assuntos
Adulto , Feminino , Humanos , Masculino , Gravidez , Sequência de Aminoácidos , DNA Complementar , Genética , Regulação Enzimológica da Expressão Gênica , Genética , Infertilidade Masculina , Genética , Isoenzimas , Genética , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Piridoxal Quinase , Genética , Splicing de RNA , Genética , RNA Mensageiro , Genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células de Sertoli , Metabolismo , Espermatogênese , Genética , Espermatozoides , Metabolismo , Testículo , Embriologia , Distribuição TecidualRESUMO
The use of fluorescent compounds can be a valuable tool to probe the active site of enzymes. Several examples of this approach are discussed, particularly the use of pyridoxal phosphate analogs. The study of protein-protein interactions by means of fluorescent-labeled proteins is also analyzed
Assuntos
Nucleotídeos de Adenina/química , Sítios de Ligação , Piridoxal Quinase/química , Piridoxal/química , Piridoxina/química , Transaminases/química , Anisotropia , Conformação ProteicaRESUMO
Säo apresentados dois casos de herpes gestacional, com uma revisäo da literatura. Embora a incidência seja baixa, a doença é extremamente desagradável, por produzir intenso prurido, pelo aspecto repulsivo e por poder atravessar a placenta e atingir o feto. Sua etiologia é uma condiçäo auto-imune e seu quadro clínico muito parecido com o do penfigóide bolhoso, com o qual deve ser feito o diagnóstico diferencial, o que só é possível com a análise ultra-estrutural da membrana basal. No tratamento, as drogas de escolha säo os corticoesteróides, seguidos de piridoxina, e é citada a troca de plasma materno. Antihistamínicos, corticoesteróides tópicos e loçöes antipuriginosas säo de açäo limitada. Descrevem-se os dois casos vistos pelos autores, mas apenas um feto foi avaliado. Este concepto näo apresentou qualquer lesäo herpética ou malformaçäo ao nascer