RESUMO
The aim of the current study was to evaluate the levels of growth factors in the cerebrospinal fluid (CSF) of patients with autism, after transplantation of human umbilical cord blood mononuclear cells (CBMNCs) and umbilical cord-derived mesenchymal stem cells (UCMSCs). Twenty patients received two CBMNC intravenous and intrathecal infusions, each followed by two UCMSC intrathecal injections. A 2-mL sample of CSF was taken before each intrathecal injection. CSF levels of hepatocyte growth factor (HGF), brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF) and basic fibroblast growth factor (bFGF) were determined by an enzyme-linked immunosorbent assay (ELISA). All data are reported as means ± SD and were analyzed using the SPSS 10.0 software. One-way analysis of variance with post-hoc F- and Q-tests was performed for comparison. HGF, BDNF and NGF levels in the CSF were significantly increased after transplantation (P < 0.05), while bFGF levels did not change significantly. Therefore, transplantation of CBMNCs and UCMSCs could increase HGF, BDNF and NGF levels in the CSF of patients with autism.
RESUMO
The aim of this study was to establish a metastatic human neuroblastoma (NB) mouse model by xenograft in order to study the metastatic mechanisms of NB. A human NB cell line was obtained from a 5-year-old patient and cultured in vitro. A suspension of these cells was subcutaneously inoculated into nude mice at the right flank next to the forelimb. The biological characteristics of the developed subcutaneous and metastatic tumors were analyzed by hematoxylin and eosin staining. The expression of the tumor marker neuron-specific enolase was determined by immunohistochemistry, and the invasive ability of metastatic tumors was examined by a Matrigel invasion assay. DNA microarray analyses were performed to examine the metastasis-related gene expression. Our results showed that tumors grew in 75% of the mice injected with NB cells and the rate of metastasis was 21%. The xenograft tumors retained the morphological and biological characteristics of the NB specimen from the pediatric patient. Neuron-specific enolase was highly expressed in both subcutaneous and metastatic tumors. The metastatic tumor cells possessed a higher invasive capability than the primary NB cells. The expression of 25 metastasis-related genes was found to be significantly altered in metastatic tumors compared to primary tumors, including RECK, MMP2, VEGF, MMP3, and CXCL12. In conclusion, we successfully established a human NB xenograft model with high tumor-bearing and metastatic rates in nude mice, providing an ideal animal model for the in vivo study of NB.
Assuntos
Neuroblastoma/patologia , Animais , Biomarcadores Tumorais , Biópsia , Linhagem Celular Tumoral , Pré-Escolar , Modelos Animais de Doenças , Feminino , Expressão Gênica , Xenoenxertos , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Nus , Metástase Neoplásica , Neuroblastoma/genética , Neuroblastoma/metabolismo , Fosfopiruvato Hidratase/genética , Fosfopiruvato Hidratase/metabolismoRESUMO
Neuroblastoma is the most common and one of the deadliest among pediatric tumors; however, a subset of infants with neuroblastoma display spontaneous regression. The mechanism of spontaneous regression remains to be elucidated. TrkA plays an essential role in the differentiation and functionality of neurons; abundant TrkA expression is associated with favorable prognosis of neuroblastoma. All-trans retinoic acid (ATRA), a first-line drug for acute promyelocytic leukemia (APL) treatment, has been shown to induce differentiation and inhibit cell growth. Neuroblastoma tissues in our hospital inpatient were collected, primary cell culture was performed, and the cells were separated and purified to be cell line. Trypan blue exclusion was used to count the numbers of cells alive, morphological changes were observed under the phase-contrast microscope. RT-PCR was used to determine the expression level of TrkA. In this study, a human neuroblastoma cell line was successfully established; in addition, we demonstrated that ATRA induces growth arrest and promotes the differentiation of neuroblastoma cells. In addition, ATRA was shown to significantly increase the levels of TrkA mRNA expression. Therefore, we concluded that the elevated expression of the TrkA receptor is associated with ATRA-induced growth arrest and differentiation o neuroblastoma cells. The results of this study provide a theoretical basis for the clinical application of differentiation-inducing ATRA for neuroblastoma therapy.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Expressão Gênica , Neuroblastoma/genética , Neuroblastoma/patologia , Receptor trkA/genética , Tretinoína/farmacologia , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular Tumoral , Humanos , RNA Mensageiro/genéticaRESUMO
We aimed to evaluate the levels of growth factors in the cerebrospinal fluid (CSF) of patients with autism after transplantation of umbilical cord blood mononuclear cells (CBMNCs). Fourteen subjects diagnosed with autism received transplantation of CBMNCs first through intravenous infusion, and three times subsequently through intrathecal injections. A 2-mL sample of CSF was taken before each intrathecal injection. CSF levels of nerve growth factor (NGF), vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) were determined by enzyme-linked immunosorbent assay. All data are reported as means ± SD and were analyzed using the SPSS 10.0 software. One-way analysis of variance with post-hoc F-and Q-tests were performed for comparisons. NGF levels in the CSF were significantly increased after transplantation (213.54 ± 56.38 after the third versus 28.32 ± 12.22 ng/L after the first transplantation; P < 0.05), while VEGF and bFGF levels did not change significantly. Therefore, transplantation of CBMNCs could increase NGF levels in the CSF of patients with autism.
Assuntos
Transtorno Autístico/líquido cefalorraquidiano , Sangue Fetal/citologia , Leucócitos Mononucleares/transplante , Fator de Crescimento Neural/líquido cefalorraquidiano , Transtorno Autístico/terapia , Criança , Pré-Escolar , Feminino , Fator 2 de Crescimento de Fibroblastos/líquido cefalorraquidiano , Humanos , Masculino , Resultado do Tratamento , Fator A de Crescimento do Endotélio Vascular/líquido cefalorraquidianoRESUMO
This study investigated the changes in peripheral benzodiazepine receptors (PBRs) in the penumbra after cerebral ischemia-reperfusion injury, and examined the effects of astragaloside IV (AST) on PBRs in rats. Sixty Sprague-Dawley rats were divided into a sham operation group, a model group, and three AST treatment groups. Cerebral ischemic models were induced by the clue-blocked method. Neurological deficits were examined. The animals were sacrificed after 2 h of ischemia and 24 h of reperfusion, and mitochondria from the penumbra were purified. PBR density (Bmax) and affinity were measured by radioligand assays. Mitochondrial [(3)H]PK11195 binding was correlated with neurological deficits in rats. Compared to the model group, the 10 mg/kg AST group, 40 mg/kg AST group, and 100 mg/kg AST group had fewer neurological deficits. The effects in the 40 mg/ kg group did not significantly differ from the effects in the 100 mg/ kg group. Compared to the model group, the 10 mg/kg AST group, 40 mg/kg group, and 100 mg/kg group had a decreased Bmax in the penumbra. The Bmax decreased in the 40 mg/kg AST group and in the 100 mg/kg AST group compared with the 10 mg/kg group. The Bmax and neurological deficits in the 40 mg/kg did not significantly differ from those in the 100 mg/kg group. By contrast, the AST-treated rats showed no significant changes in the binding parameter equilibrium dissociation constant compared with those in the sham operation group and the model group. AST protects ischemic brain tissue by inhibiting PBR expression after cerebral ischemia.