OBJECTIVETo investigate the protective effect of ginsenoside Rg1 on oxygen-glucose deprivation (OGD) in PC-12 cells, and preliminarily discuss the potential molecular mechanism of mTOR/Akt/FoxO3 signaling pathway.

METHODThe OGD PC-12 cell model was established. The cell viability was measured by MTT assay. After the pretreatment with Rg1 with the concentration of 10, 20, 40 micromol x L(-1) for 24 h, the cell viability was observed. Lactate dehydrogenase (LDH) release, superoxide dismutase (SOD) ac- tivity and malondialdehyde (MDA) level were detected by colorimetry assay. mTOR, p-Akt(ser473), p-Akt(tjr308), Akt, p-FoxO3, FoxO3 in cytoplasm and nucleus, and total FoxO3 protein expression were detected by Western blot assay.

RESULTOGD could significantly in- hibit cell proliferation in 4-24 h in a time-dependent manner. After pretreatment for 24 h, Rg1 (20, 40 micromol x L(-1)) could notably elevate the cell viability and SOD viability and reduce the LDH release and MDA content. Besides, Rg1 also inhibited OGD-induced mTOR and p-Akt(ser473) decreases. After treatment for 6 h, OGD could reduce FoxO3 phosphorylation and promote FoxO3 in cytoplasm. This data suggested that Rg1 could protect PC-12 cell injury through mTOR/p-Akt/FoxO3 signaling pathway.

CONCLUSIONGinsenoside Rg1 could attenuate OGD-induced PC-12 cell injury. Its action mechanism may be closely related to activation of mTOR/p-Akt/FoxO3 signaling pathway.

Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Forkhead Box Protein O3 ; Forkhead Transcription Factors ; genetics ; metabolism ; Ginsenosides ; pharmacology ; Glucose ; metabolism ; Oxygen ; metabolism ; PC12 Cells ; Protective Agents ; pharmacology ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Rats ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

According to published nucleotide sequences, ORF4 gene of barley yellow dwarf virus GAV (BYDV-GAV) was synthesized by reverse transcription-polymerase chain reaction (RT-PCR). The BYDV-GAV ORF4 gene was expressed in baculovirus -insect cell expression system efficiently, and western bolt analysis confirmed its expression product. Confocal laser scanning microscopy showed that GFP: ORF4 fusion protein was associated with the nuclear envelope of insect cells. By expressing the N- and C-terminal regions of ORF4-encoding product (P4) in insect cells combined with structure prediction, it was found that the N-terminal region of P4 containing four a-helices is required for targeting P4 to the nuclear envelope. These results provide a base for biological function of ORF4 gene during systemic infection of BYDV-GAV in host plants further.
Amino Acid Sequence ; Animals ; Baculoviridae ; genetics ; metabolism ; Genes, Plant ; genetics ; Green Fluorescent Proteins ; genetics ; Insecta ; genetics ; metabolism ; Luteovirus ; genetics ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; metabolism

OBJECTIVETo detect the sequence variations frequently found within the N- and C-terminal regions of Epstein-Barr virus (EBV) LMP1 gene in nasopharyngeal carcinoma (NPC) and to study the underlying mechanisms.

METHODSFresh tumor tissues were sampled from 63 patients with untreated NPC encountered in Affiliated Tumor Hospital of Sun Yat-sen University, Guangzhou. The N-terminal region of EBV LMP1 gene was amplified with nested polymerase chain reaction (PCR), followed by XhoI enzyme digestion. Nested PCR was also employed to detect the 30 base pairs deletion within the C-terminal region. Four-colored fluorescence terminator sequencing method was applied for bi-directional solid-phase sequencing of the 8 representative PCR products in 4 cases of NPC. The DNA sequence within the N- and C-terminal regions of LMP1 gene was then analyzed.

RESULTSThere were 4 patterns of sequence variations, namely, wt-XhoI/wt-LMP1 (4 cases, 6.3%), wt-XhoI and XhoI-loss/del-LMP1 (4 cases, 6.3%), wt-XhoI/del-LMP1 (5 cases, 7.9%) and XhoI-loss/del-LMP1 (50 cases, 79.5%), detected in the 63 studied cases. Sequence analysis showed that the EBV LMP1 gene had underwent non-synonymous and synonymous substitutions, as compared with the prototype of B95-8 cells. The ratio of non-synonymous to synonymous substitutions was 2.25.

CONCLUSIONSXhoI-loss/del-LMP1 is the predominant sequence variation pattern of EBV LMP1 gene in NPC from Guangzhou. The XhoI-loss variation seems to develop on top of del-LMP1. When compared with the EBV LMP1 gene in peripheral blood B-lymphocytes of virus carriers and in preinvasive epithelial lesions (reported previously), it is likely that the sequence variation patterns of LMP1 gene may represent 4 different phases of intrahost evolution of EBV during nasopharyngeal carcinogenesis.

Adult ; Aged ; Base Sequence ; DNA, Viral ; genetics ; Deoxyribonucleases, Type II Site-Specific ; genetics ; Female ; Gene Deletion ; Genetic Variation ; Herpesvirus 4, Human ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Nasopharyngeal Neoplasms ; virology ; Point Mutation ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

In order to discover light quality's effects on growth, photosynthesis and effective components content of Panax notoginseng, a pot experiment using 7 light qualities (red, orange, yellow, green, cyan, violet, and blue) was conducted. The growth, photosynthesis and content change of effective components were measured during plant growth. The results showed that light qualities had significant effect on plant growth, red light increased the plant height, while cyan, yellow, violet, and blue lights promoted accumulation of biomass underground, blue and yellow lights increased the photosynthesis, cyan light increased accumulation of ginsenoside Rd, yellow and cyan lights increased total effective components of individual plant.
Light ; Panax notoginseng ; growth & development ; metabolism ; radiation effects ; Photosynthesis ; radiation effects ; Plant Extracts ; analysis ; metabolism

OBJECTIVETo develop the mechanism of improving protection function of prepared Cornus officinalis for liver and kidney and the biological activity of 5-hydroxymethylfurfural (5-HMF).

METHODPharmacological and chemical studies were used to choose active part. A compound from active part was separated and appraised. To investigate his biological functions, pharmacological experiment was actualized.

RESULTA component was separated and identified. His is 5-HMF. 5-HMF can protect human vein epidermal cell against H2O2 and glucose and inprove acute liver injury in mice.

CONCLUSION5-HMF is the active component in prepared Cornus officinalis and substance basis for protecting liver and kidney.

Animals ; Cells, Cultured ; Cornus ; chemistry ; Endothelial Cells ; drug effects ; Female ; Furaldehyde ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Liver ; drug effects ; Male ; Mice ; Random Allocation

Objective　To study the effects of measures for preventing early postburn damage in improving survival rate of burn patients during the third stage. Methods　12 568 burn cases admitted to our institute were chronically divided into three groups (1958-1980;1981-1990;1991-2000). Total burn surface area (TBSA), survival rate, incidence of burn shock, systemic infection and organ damage as well as the main treatments adopted in the recent decade were retrospectively analyzed. Results　Incidence of burn shock, systemic infection and organ damage were significantly lower, and the total survival rate and the survival rate in patients with different TBSA were markedly higher in the third group as compared with those in the first and the second group. Incidence of organ damage in patients treated with delayed fast fluid infusion, early escharectomy en masse, early enteral feeding, early prevention of inhalation injury and gut bacterial translocation were also significantly lower than in the control. Conclusion Measures taken in the third group for preventing early postburn damage play an important role in improving the survival rate of burn patients.

Blotting, Western ; DNA, Ribosomal ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; biosynthesis ; Transfection ; Vascular Endothelial Growth Factor A ; genetics

BACKGROUNDFamilial hypercholesterolemia (FH) is a type of dominant autosomal disease that causes high levels of plasma low-density lipoprotein cholesterol (LDL-C). In the past years, molecular data related to FH were limited in China. Now, to gain more information about FH, we analyzed one proband with a severe FH phenotype as well as his relatives.

METHODSAfter the entire coding sequence and the intron-exon junctions of the low-density lipoprotein receptor (LDLR) gene were amplified using PCR, we sequenced the LDLR gene of a Chinese FH family. RT-PCR was used to detect changes in the mRNA.

RESULTSTwo novel mutations were identified in the LDLR gene of this family. One, W165X, was a G > A substitution at the third nucleotide of codon 165. The other, IVS5-1G > A, was also a G > A substitution at the acceptor splice site of intron 5. The most striking discovery is that the proband was heterozygous for W165X but homozygous for IVS5-1G > A. The cDNA sequencing showed that the IVS5-1G > A mutation caused the insertion of 10 nucleotides, namely GCTCTCACAA, between exon 5 and exon 6.

CONCLUSIONSThe two nucleotide variations are thought to be the FH-causing mutations because the co-segregation of the mutant allele with the phenotype of FH has been shown in this Chinese family. These data show an increase in the mutational spectrum of FH in China and verify a scarce mutational form in the LDLR gene.

Adult ; Child ; DNA, Complementary ; analysis ; Female ; Humans ; Hyperlipoproteinemia Type II ; genetics ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Receptors, LDL ; genetics

OBJECTIVETo construct the prokaryotic plasmid of FUS1 gene for efficient FUS1 expression in E.coli strain Rosetta(DE3)2plys.

METHODSThe full-length FUS1 gene was amplified by PCR from the total RNA of umbilical mesenchymal stem cells and cloned into pET-32a(+) vector followed by identification with PCR and sequencing. The recombinant plasmid pET-32a(+)-FUS1 was transformed into the E.coli strain Rosetta(DE3)2plys and the target protein expression was induced by IPTG.

RESULTSThe plasmid pET-32a(+)-FUS1 was obtained successfully as verified by PCR and sequence analysis. High expression of the fused FUS1 protein was achieved after induction by low-concentration IPTG (25 micromol/L) for 3 h, and the recombinant FUS1 protein accounted for 40% of the total bacterial protein of Rosetta(DE3)2plys.

CONCLUSIONThe recombinant FUS1 plasmid has been successfully cloned, which allows highly efficient FUS1 expression in Rosetta (DE3)2 plys.

Blotting, Western ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; Transformation, Genetic ; Tumor Suppressor Proteins ; biosynthesis ; genetics ; Umbilical Cord ; cytology