BACKGROUNDAdipose tissue-derived stromal cells (ADSCs) can be greatly expanded in vitro, and induced to differentiate into multiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic, and adipogenic cells. This study was designed to investigate the possibility of ADSCs differentiating into neurons.

METHODSAdipose tissue from rats was digested with collagenase, and adherent stromal cells were cultured. A medium containing a low concentration of fetal bovine serum was adopted to induce the cells to differentiate. ADSCs were identified by immunocytochemistry, and semi-quantitative RT-PCR was applied to detect mRNA expression of neurofilament 1 (NF1), nestin, and neuron-specific enolase (NSE).

RESULTSNestin-positive cells were found occasionally among ADSCs. ADSCs were found to express NSE mRNA and nestin mRNA, but not NF1 mRNA. ADSCs could differentiate into neuron-like cells in a medium composed of a low concentration of fetal bovine serum, and these differentiated cells displayed complicated neuron-like morphologies.

CONCLUSIONSThe data support the hypothesis that adipose tissue contains stem cells capable of differentiating into neurons. These stem cells can overcome their mesenchymal commitment, and may represent an alternative autologous stem cell source for CNS cell transplantation.

Adipose Tissue ; cytology ; Animals ; Cell Differentiation ; physiology ; Cells, Cultured ; Immunohistochemistry ; Intermediate Filament Proteins ; analysis ; Nerve Tissue Proteins ; Nestin ; Neurofilament Proteins ; analysis ; Neurons ; cytology ; Phenotype ; Phosphopyruvate Hydratase ; analysis ; Rats

OBJECTIVETo investigate the effects of preweaning enrichment on the expression of activity-regulated cytoskeletal protein (Arc), an immediate early gene, and on the long-term memory in rats.

METHODSForty neonatal Sprague-Dawley rats were randomly assigned to control group (standard environment, n=20) and experimental group (enriched environment, n=20). The experimental group received enriched environment exposure from postnatal day 10 until weaning (2 weeks, 20 minutes per day). The open field and novel object recognition tests were performed at postnatal day 28. Arc expression was detected by Western blotting and immunohistochemistry.

RESULTSThere was no significant difference in the open field test between the two groups. However, in the novel object recognition test, the experimental group rats performed significantly better than the control rats after 1 and 24-hr retention. The preference index in the experimental group after 1-hr (59.61%+/-9.61% vs 50.46%+/-9.34%; P<0.05) and 24-hr retention (62.72%+/-14.12% vs 52.39%+/-9.16%; P<0.05 ) was significantly higher than that in the control group. Arc expression in both areas CA1 and DG of hippocampus in the experimental group increased significantly compared with that in the control group (P<0.01).

CONCLUSIONSPreweaning enrichment can up-regulate the expression of immediate early gene, Arc, in the hippocampus of the rats, and promote their long-term memory.

Animals ; Cytoskeletal Proteins ; analysis ; Female ; Hippocampus ; chemistry ; physiology ; Immunohistochemistry ; Male ; Memory ; Nerve Tissue Proteins ; analysis ; Rats ; Rats, Sprague-Dawley

Humans ; Hypertension ; genetics ; Multivariate Analysis ; Nerve Tissue Proteins ; genetics ; Polymorphism, Genetic

OBJECTIVETo observe the expression and distribution of adult rat axon guidance cues Netrin-1 and Slit2 at different time points after spinal cord injury and to investigate the guidance mechanism of regenerated axons.

METHODSTwenty adult Sprague Dawley (SD) rats were divided randomly into five groups with 4 in each. Four groups of them were used to make Allen's spinal cord punch models and we took materials randomly from one of them on the 2nd, 4th, 7th and 14th day respectively after operation. The left one group was taken as the control group. Immunofluorescence laser confocal scan was used to examine the co-expression and localization of Netrin-1 and Slit2 proteins in the injured site of the spinal cord.

RESULTSWithin two weeks after SCI, the expression of Netrin-1 and Slit2 proteins increased temporarily and there was co-expression of them on the neuron plasma membrane.

CONCLUSIONSSynchronous high expression and co-expression of axon attractant Netrin-1 and repellent Slit2 are found in the adult rat injured spinal cord in the damaged local and vicinity parts, and probably, they act as the key regulators of axon guidance regeneration.

Animals ; Female ; Intercellular Signaling Peptides and Proteins ; analysis ; Male ; Microscopy, Confocal ; Nerve Growth Factors ; analysis ; Nerve Regeneration ; physiology ; Nerve Tissue Proteins ; analysis ; Netrin-1 ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; metabolism ; Tumor Suppressor Proteins ; analysis

OBJECTIVETo investigate the clinical features and proline-rich transmembrane protein 2 (PRRT2) gene mutation in patients with paroxysmal kinesigenic dyskinesia (PKD).

METHODClinical information was collected at Peking University First Hospital from January 2004 to July 2014. In total, 10 patients with PKD were recruited, and all were males. Among them, four patients were the probands from four PKD families and the other six patients were sporadic cases. Clinical information was analyzed. Peripheral blood samples for DNA study were collected from PKD patients and their family members. Genomic DNA was extracted using standard procedures. Mutation analysis of PRRT2 was performed by Sanger sequencing after PCR.

RESULTOf the 10 patients, the median age of dyskinesias onset was 10 years, ranging from 4 to 13 years. The description of their attacks were abnormal involuntary movements provoked by sudden movements, without loss of consciousness. Five patients exhibited dystonia, two patients exhibited choreoathetosis, and three patients had mixed (dystonia and choreoathetosis) dyskinesias. The duration of the attacks lasted for 3 to 30 seconds. The frequency ranged from once per month to twenty times per day. PRRT2 mutations, c. 649_650insC (p. R217PfsX8), were found in all the four PKD families. Mutation c. 649_650insC was also detected in two of the six sporadic PKD cases, inheriting from their asymptomatic mother.

CONCLUSIONThe onset age of PKD could be in the early childhood. The clinical features of the familial cases and sporadic cases showed no difference. The attacks manifested as dystonia, choreathetosis, or mixed. PRR2 mutations could be identified in familial or sporadic cases with PKD. Mutation c. 649_650insC is the hotspot mutation of PRRT2 gene.

Adolescent ; Child ; Child, Preschool ; DNA Mutational Analysis ; Dystonia ; genetics ; Humans ; Male ; Membrane Proteins ; genetics ; Mutation ; Nerve Tissue Proteins ; genetics

BACKGROUNDMany researchers suggest that adult mammalian central nervous system (CNS) is incapable of completing self-repair or regeneration. And there are accumulating lines of evidence which suggest that endogenous neural stem cells (NSCs) are activated in many pathological conditions, including stroke in the past decades, which might partly account for rehabilitation afterwards. In this study, we investigated whether there was endogenous neural stem cell activation in intracerebral hemorrhagic (ICH) rat brains.

METHODSAfter ICH induction by stereotactical injection of collagenase type VII into globus pallidus, 5-Bromo-2 Deoxyuridine (BrdU) was administered intraperitoneally to label newborn cells. Immunohistochemical method was used to detect Nestin, a marker for neural stem cells, and BrdU.

RESULTSNestin-positive or BrdU-Labeled cells were predominantly located at 2 sites: basal ganglion around hemotoma, ependyma and nearby subventricular zone (SVZ). No positive cells for the 2 markers were found in the 2 sites of normal control group and sham group, as well as in non-leisioned parenchyma, both hippocampi and olfactory bulbs in the 4 groups. Nestin+ cells presented 4 types of morphology, and BrdU+ nucleus were polymorphologic. Positive cell counting around hemotoma showed that at day 2, Nestin+ cells were seen around hemotoma in model group, the number of which increased at day 4, day 7 (P <0.01), peaked at day 14 (P <0.05), and reduced significantly by day 28 (P <0.01).

CONCLUSIONEndogenous neural stem cells were activated in experimental intracerebral hemorrhagic rat brains.

Animals ; Brain ; pathology ; Bromodeoxyuridine ; metabolism ; Cerebral Hemorrhage ; pathology ; Intermediate Filament Proteins ; analysis ; Male ; Nerve Tissue Proteins ; analysis ; Nestin ; Neurons ; pathology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; pathology

OBJECTIVETo investigate the expression of nesfatin-1/NUCB2 and ghrelin in the gastric mucosa of rats with intrauterine growth retardation (IUGR) and its significance.

METHODSThe IUGR animal model was established by feeding rats low-protein diets during their pregnancy. Newborn rats were divided into catch-up growth, non-catch-up growth and control groups. Protein and mRNA levels of nesfatin-1/NUCB2 and ghrelin in the gastric mucosa of rats were determined by RT-PCR and Western blot, respectively.

RESULTSNesfatin-1/NUCB2 mRNA and protein were expressed in the gastric mucosa of rats immediately after birth, and their expression increased in an age-dependent manner in all three groups. Furthermore, the level of nesfatin-1/NUCB2 in the catch-up growth group was higher than that in the control group before weaning, whereas there was no significant difference in nesfatin-1/NUCB2 expression between the two groups after weaning. The level of nesfatin-1/NUCB2 in the non-catch-up growth group was lower than that in the catch-up growth group during the whole observation period. The level of ghrelin in the catch-up growth group was higher than that in the control group starting from day 12 after birth, whereas there was no significant difference in ghrelin expression between the two groups after weaning. The level of ghrelin in the non-catch-up growth group was lower compared with those in the catch-up growth and control groups from days 12 to 28 after birth.

CONCLUSIONSNesfatin-1 and ghrelin are co-expressed in the gastric mucosa of rats with IUGR after birth and interact with each other to produce long-term nutritional regulation.

Age Factors ; Animals ; Calcium-Binding Proteins ; analysis ; genetics ; DNA-Binding Proteins ; analysis ; genetics ; Female ; Fetal Growth Retardation ; metabolism ; Gastric Mucosa ; chemistry ; Ghrelin ; analysis ; genetics ; Male ; Nerve Tissue Proteins ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley

A 5-year-old, male French bulldog with bradycardia, dyspnea, and decerebrate rigidity was necropsied. Macroscopic findings were restricted to the brain, and a single mass, 1.5x2.0x1.5 cm in size, was observed mainly at the right cingulum with prominently protruding into the dilated right lateral ventricle. The mass was grayish white in color, soft and gelatinous, but not clearly delineated. Microscopically, the mass consisted of diffuse proliferated neoplastic oligodendroglial cells characterized by small, round, and hyperchromatic nuclei with clear cytoplasm and the cells aggressively invaded into the adjacent parenchyma. Immunohistochemistry demonstrated that most of the neoplastic cells were positive for S-100 protein, vimentin, neuron specific enolase (NSE), and neurofilament protein (NFP). From these findings, the mass was diagnosed as oligodendroglioma.
Animals ; Cerebral Ventricle Neoplasms/pathology/*veterinary ; Dog Diseases/*pathology ; Dogs ; Immunohistochemistry ; Male ; Nerve Tissue Proteins/analysis ; Oligodendroglioma/pathology/*veterinary

OBJECTIVETo develop an optimal sequencing system which can improve the resolution of sequencing G-C rich DNA with abundant trinucleotide repeats by applying concentration gradients of betaine to the Sanger sequencing system.

METHODSConcentration gradients of betaine were introduced into the sequencing system by taking the 5' terminal of Nogo-B cDNA (Am-Nogo-B) (G-C%=72%, without trinucleotide repeats) and 5' terminal of Huntingtin cDNA (Am-HTT) (G-C%=74%, with abundant CAG and CCG repeats) the results of sequencing were compared.

RESULTSThe optimum concentration of betaine for sequencing Am-Nogo-B has differed from that for Am-HTT. Result of sequencing Am-Nogo-B has achieved the best quality when the concentration of betaine was at 0.8-1.2 mol/L, whereas the result of sequencing Am-HTT obtained the best quality when the concentration of betaine was at 1.6 -2.4 mol/L. The results were reproducible.

CONCLUSIONG-C rich DNA with similar G-C% required different concentrations of betaine in the sequencing system due to base pair compositions. The sequencing system developed for improving the resolution of sequencing of G-C rich DNA with abundant trinucleotide repeats can be used as a reference for similar studies.

Base Sequence ; Betaine ; pharmacology ; Huntingtin Protein ; Molecular Sequence Data ; Nerve Tissue Proteins ; genetics ; Sequence Analysis, DNA ; methods ; Trinucleotide Repeats

Objectives: It is unclear whether G protein-coupled receptor 61 (GPR61) affecting body weight, plays a role in the association between birth weight and weather. This study aimed to assess the effects of prenatal weather and GPR61 on birth weight. Methods: A total of 567 mother-newborn pairs were recruited in Houzhai Center Hospital during 2011-2012. We detected the maternal and neonatal GPR61 promoter methylation levels, and obtained meteorological and air pollution data. Results: A positive association was observed between maternal and neonatal GPR61 methylation levels, and both of them were affected by precipitation, relative humidity (RH) and daily temperature range (DTR). Birth weight was associated negatively with RH and positively with DTR ( P < 0.05). A significant association was observed between birth weight and neonatal GPR61 methylation. We observed that maternal GPR61 methylation seemed to modify associations between weather and birth weight ( P _interaction < 0.10), while neonatal GPR61 methylation mediated the effects of RH and DTR on birth weight ( P < 0.05). Conclusions Our findings revealed the significant associations among prenatal weather, GPR61 methylation and birth weight. Maternal GPR61 methylation may modify the susceptibility of birth weight to prenatal weather conditions, while neonatal GPR61 methylation may be a bridge of the effects of prenatal RH and DTR on birth weight.
Air Pollution/analysis* ; Birth Weight ; Female ; Humans ; Infant, Newborn ; Nerve Tissue Proteins ; Pregnancy ; Receptors, G-Protein-Coupled/metabolism* ; Temperature ; Weather