Objective To explore the repaired effects of autotransplantation of bone marrow-derived neural stem cells (BMSCs) on hippocampus of epilepsy rats.Methods Male SD rats were randomly divided into groups normal control (NC),graft and non-graft .Under sterile condition, the BMSCs of rats were isolated, and under specific condition the BMSCs were cultured to induce and differentiate into neural stem cells(NSCs). Then the models of temporal lobe epilepsy were established in groups graft and non-graft , and the NSCs were autotransplanted into the right hippocampus of rats of graft group. The morphological changes of the hippocampus were observed at 1,2 ,4,8,16 weeks after the transplantation respectively.Results The number of hippocampal CA3 pyramid cells of groups non-graft and graft significantly decreased than that in NC group(all P

Objective:To examine differences in metabolic characteristics and metabolites between elderly overweight patients with metabolic syndrome and healthy elderly people, and to identify related factors.Methods:A group of 36 MS patients(the MS group)admitted to The Fourth Central Hospital of Tianjin from April to August 2018 and 43 elderly people(the control group)who underwent physical examination during the same period were included in this prospective study.Serum samples of the patients with metabolic syndrome and elderly healthy controls were collected, and ultra-performance liquid chromatography and quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)based non-targeted metabolomics was used to search for differences in metabolites between the serum samples of the two groups.The Pearson correlation statistical method was used to find related clinical factors.Results:Comparison of baseline data of the enrolled participants showed that there were statistically significant differences between the two groups in body mass index[(26.9±2.0)kg/m 2vs.(21.7±1.4)kg/m 2], waist circumference, systolic blood pressure, fasting blood glucose, triglycerides and high-density lipoprotein cholesterol( P<0.01). Metabolomics results showed that there were differences in 65 serum metabolites between elderly overweight patients with metabolic syndrome and elderly normal controls, and these differences were enriched in 21 pathways.Correlation analysis showed that waist circumference had the largest number of differential metabolites, followed by body mass index.The major differential metabolites were monosaccharides such as mannose, lyxose and glucose, linolenic acid and its derivatives, and pyroglutamate. Conclusions:Compared with normal elderly people, elderly patients with overweight metabolic syndrome have a variety of differential metabolites, and these metabolites are highly correlated with clinical indicators related to overweight, such as body mass index and waist circumference, and they include monosaccharides, linolenic acid derivatives and amino acids.

Knowledge of the evolution of pathogens is of great medical and biological significance to the prevention, diagnosis, and therapy of infectious diseases. In order to understand the origin and evolution of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus), we collected complete genome sequences of all viruses available in GenBank, and made comparative analyses with the SARS-CoV. Genomic signature analysis demonstrates that the coronaviruses all take the TGTT as their richest tetranucleotide except the SARS-CoV. A detailed analysis of the forty-two complete SARS-CoV genome sequences revealed the existence of two distinct genotypes, and showed that these isolates could be classified into four groups. Our manual analysis of the BLASTN results demonstrates that the HE (hemagglutinin-esterase) gene exists in the SARS-CoV, and many mutations made it unfamiliar to us.
Amino Acid Motifs ; Amino Acid Substitution ; Base Composition ; Codon ; genetics ; Computational Biology ; DNA Mutational Analysis ; Evolution, Molecular ; Gene Transfer, Horizontal ; Genetic Variation ; Genome, Viral ; Phylogeny ; SARS Virus ; genetics

The R (replicase) protein is the uniquely defined non-structural protein (NSP) responsible for RNA replication, mutation rate or fidelity, regulation of transcription in coronaviruses and many other ssRNA viruses. Based on our complete genome sequences of four isolates (BJ01-BJ04) of SARS-CoV from Beijing, China, we analyzed the structure and predicted functions of the R protein in comparison with 13 other isolates of SARS-CoV and 6 other coronaviruses. The entire ORF (open-reading frame) encodes for two major enzyme activities, RNA-dependent RNA polymerase (RdRp) and proteinase activities. The R polyprotein undergoes a complex proteolytic process to produce 15 function-related peptides. A hydrophobic domain (HOD) and a hydrophilic domain (HID) are newly identified within NSP1. The substitution rate of the R protein is close to the average of the SARS-CoV genome. The functional domains in all NSPs of the R protein give different phylogenetic results that suggest their different mutation rate under selective pressure. Eleven highly conserved regions in RdRp and twelve cleavage sites by 3CLP (chymotrypsin-like protein) have been identified as potential drug targets. Findings suggest that it is possible to obtain information about the phylogeny of SARS-CoV, as well as potential tools for drug design, genotyping and diagnostics of SARS.
Amino Acid Sequence ; Base Composition ; Base Sequence ; Cluster Analysis ; Computational Biology ; Conserved Sequence ; genetics ; Evolution, Molecular ; Gene Components ; Genome, Viral ; Molecular Sequence Data ; Mutation ; genetics ; Phylogeny ; Protein Structure, Tertiary ; RNA Replicase ; genetics ; SARS Virus ; genetics ; Sequence Analysis, DNA

Expressed Sequence Tag (EST) analysis has pioneered genome-wide gene discovery and expression profiling. In order to establish a gene expression index in the rice cultivar indica, we sequenced and analyzed 86,136 ESTs from nine rice cDNA libraries from the super hybrid cultivar LYP9 and its parental cultivars. We assembled these ESTs into 13,232 contigs and leave 8,976 singletons. Overall, 7,497 sequences were found similar to existing sequences in GenBank and 14,711 are novel. These sequences are classified by molecular function, biological process and pathways according to the Gene Ontology. We compared our sequenced ESTs with the publicly available 95,000 ESTs from japonica, and found little sequence variation, despite the large difference between genome sequences. We then assembled the combined 173,000 rice ESTs for further analysis. Using the pooled ESTs, we compared gene expression in metabolism pathway between rice and Arabidopsis according to KEGG. We further profiled gene expression patterns in different tissues, developmental stages, and in a conditional sterile mutant, after checking the libraries are comparable by means of sequence coverage. We also identified some possible library specific genes and a number of enzymes and transcription factors that contribute to rice development.
Arabidopsis ; genetics ; DNA, Complementary ; metabolism ; Databases as Topic ; Expressed Sequence Tags ; Gene Library ; Genome, Plant ; Genomics ; methods ; Multigene Family ; Open Reading Frames ; Oryza ; genetics ; Quality Control ; Software

Annotation of the genome sequence of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) is indispensable to understand its evolution and pathogenesis. We have performed a full annotation of the SARS-CoV genome sequences by using annotation programs publicly available or developed by ourselves. Totally, 21 open reading frames (ORFs) of genes or putative uncharacterized proteins (PUPs) were predicted. Seven PUPs had not been reported previously, and two of them were predicted to contain transmembrane regions. Eight ORFs partially overlapped with or embedded into those of known genes, revealing that the SARS-CoV genome is a small and compact one with overlapped coding regions. The most striking discovery is that an ORF locates on the minus strand. We have also annotated non-coding regions and identified the transcription regulating sequences (TRS) in the intergenic regions. The analysis of TRS supports the minus strand extending transcription mechanism of coronavirus. The SNP analysis of different isolates reveals that mutations of the sequences do not affect the prediction results of ORFs.
Amino Acid Substitution ; Base Composition ; Base Sequence ; Computational Biology ; methods ; Genome, Viral ; Isoelectric Point ; Models, Genetic ; Molecular Sequence Data ; Molecular Weight ; Open Reading Frames ; SARS Virus ; genetics ; Sequence Analysis ; Transcription, Genetic

The Coronaviridae family is characterized by a nucleocapsid that is composed of the genome RNA molecule in combination with the nucleoprotein (N protein) within a virion. The most striking physiochemical feature of the N protein of SARS-CoV is that it is a typical basic protein with a high predicted pI and high hydrophilicity, which is consistent with its function of binding to the ribophosphate backbone of the RNA molecule. The predicted high extent of phosphorylation of the N protein on multiple candidate phosphorylation sites demonstrates that it would be related to important functions, such as RNA-binding and localization to the nucleolus of host cells. Subsequent study shows that there is an SR-rich region in the N protein and this region might be involved in the protein-protein interaction. The abundant antigenic sites predicted in the N protein, as well as experimental evidence with synthesized polypeptides, indicate that the N protein is one of the major antigens of the SARS-CoV. Compared with other viral structural proteins, the low variation rate of the N protein with regards to its size suggests its importance to the survival of the virus.
Amino Acid Motifs ; genetics ; Amino Acid Sequence ; Antigens, Viral ; immunology ; Base Composition ; Base Sequence ; Cluster Analysis ; Computational Biology ; DNA Primers ; Enzyme-Linked Immunosorbent Assay ; Genetic Variation ; Molecular Sequence Data ; Nucleocapsid Proteins ; genetics ; immunology ; metabolism ; Phosphorylation ; SARS Virus ; genetics ; Sequence Analysis, DNA

The E (envelope) protein is the smallest structural protein in all coronaviruses and is the only viral structural protein in which no variation has been detected. We conducted genome sequencing and phylogenetic analyses of SARS-CoV. Based on genome sequencing, we predicted the E protein is a transmembrane (TM) protein characterized by a TM region with strong hydrophobicity and alpha-helix conformation. We identified a segment (NH2-_L-Cys-A-Y-Cys-Cys-N_-COOH) in the carboxyl-terminal region of the E protein that appears to form three disulfide bonds with another segment of corresponding cysteines in the carboxyl-terminus of the S (spike) protein. These bonds point to a possible structural association between the E and S proteins. Our phylogenetic analyses of the E protein sequences in all published coronaviruses place SARS-CoV in an independent group in Coronaviridae and suggest a non-human animal origin.
Amino Acid Sequence ; Base Sequence ; Cluster Analysis ; Codon ; genetics ; Gene Components ; Genome, Viral ; Membrane Glycoproteins ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Protein Conformation ; SARS Virus ; genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology ; Spike Glycoprotein, Coronavirus ; Viral Envelope Proteins ; genetics ; metabolism

The corona-like spikes or peplomers on the surface of the virion under electronic microscope are the most striking features of coronaviruses. The S (spike) protein is the largest structural protein, with 1,255 amino acids, in the viral genome. Its structure can be divided into three regions: a long N-terminal region in the exterior, a characteristic transmembrane (TM) region, and a short C-terminus in the interior of a virion. We detected fifteen substitutions of nucleotides by comparisons with the seventeen published SARS-CoV genome sequences, eight (53.3%) of which are non-synonymous mutations leading to amino acid alternations with predicted physiochemical changes. The possible antigenic determinants of the S protein are predicted, and the result is confirmed by ELISA (enzyme-linked immunosorbent assay) with synthesized peptides. Another profound finding is that three disulfide bonds are defined at the C-terminus with the N-terminus of the E (envelope) protein, based on the typical sequence and positions, thus establishing the structural connection with these two important structural proteins, if confirmed. Phylogenetic analysis reveals several conserved regions that might be potent drug targets.
Amino Acid Sequence ; Antigens, Viral ; immunology ; Base Composition ; Computational Biology ; Enzyme-Linked Immunosorbent Assay ; Membrane Glycoproteins ; genetics ; Molecular Sequence Data ; Mutation ; genetics ; Phylogeny ; Protein Structure, Tertiary ; SARS Virus ; genetics ; immunology ; Sequence Analysis, DNA ; Sequence Homology ; Spike Glycoprotein, Coronavirus ; Viral Envelope Proteins ; genetics ; metabolism

Objective To investigate the pathogenesis of type-B aortic dissection by using morphological analysis and computational fluid dynamics (CFD) method, so as to provide evidence for the effective prediction of type-B aortic dissection. Methods Six primary type-B dissection cases scanned by CT (dissection group) and six normal cases applied to black-blood MRI (control group) were included in this study and patient-specific three-dimensional (3D) models of aorta were established through image segmentation and 3D reconstruction. The pre-type-B dissection aortas were constructed by applying the scaling algorithm to shrink the dissection and then compared with subjects in control group. The differences between morphological parameters and hemodynamic parameters of the two groups were compared. Results Compared with the normal cases, the area of the descending aorta increased dramatically in dissection group ［(892.03±263.78) mm2 vs (523.67±64.10) mm2, P=0.036］. A significant decrease in angle of the left subclavian artery occurred (66.62°±20.11° vs 100.40°±15.35°, P=0.036). The tortuosity of the aorta also had an obvious increase (0.37°±0.07° vs 0.21°±0.51°, P=0.011). The time-averaged wall shear stress (TAWSS) in dissection group was obviously higher than that in control group; the flow in the dissection region was vortex flow at low speed and the oscillating shear index (OSI) was higher. Conclusions The results of this study can be used to provide guidance for the early diagnosis and treatment of type-B aortic dissection.