Objective To investigate the effect of adenovirus-medicated gene therpy with vascular endothelial growth factor(VEGF)delivered into the subdermal space to treat compromised skin flap in the rat.Methods Thir- ty sprague-dawley rats weighing 350～400g were used in this study.The rats was randomly divided into three groups of ten rats each.A dorsal flap(8cm?2cm)in full thickness with the pedicale located at the leve of the iliaccrest was designed,each,animal received recipient bed subfascial injections of adenovirus encoding VEGF(Ad-VEGF)or green fluorescent protein(Ad-GFP)as treatment control.Another set of animals(n = 10)received NG was designated as control.The flap were elevated as originally designated and sutured back to its bed.The survival rate of the flap was evaluated on day seven after operation.Results The survival rates of the flap in Ad-VEGF group increased signifi- cantly as compared with those of the Ad-GFP group(P

OBJECTIVETo evaluate in vivo pharmacokinetics of Ophiopogonis Radix polysaccharide MDG-1 oily suspension injection prepared with different prescriptions in rats, and explore the feasibility of the long-acting drug delivery of MDG-1 Injection by using the oily suspension drug release system.

METHODMDG-1 microparticles were prepared by the anti-solvent precipitation method. Their size and size distribution were characterized. Castor oil with a high viscosity or aluminum stearate were added into soybean oil with a low viscosity, in order to prepare oily media with different viscosities, detect their rheological properties and screen out superior prescriptions for in vivo evaluation.

RESULTThe average size of microparticles was 21.81 microm, and the span between them was 2.63. The in vivo evaluation was conducted for prescriptions of mixed oil (soybean oil/castor oil, 2: 3) and soybean oils gelled by 2% and 4% aluminium stearate. Among them, the prescription of soybean gelled by 4% aluminium stearate could significantly reduce C(max) and prolong the apparent t1/2, with the MDG-1 release time of several days.

CONCLUSIONIt is feasible to achieve the long-acting MDG-1 drug delivery by using oily media with a high viscosity.

Animals ; Chemistry, Pharmaceutical ; methods ; Drug Delivery Systems ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Male ; Ophiopogon ; chemistry ; Plant Oils ; chemistry ; Polysaccharides ; administration & dosage ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Viscosity

Several kinds of column chromatography method were used to investigate the chemical constituents of the leaves of Lophatherum gracile. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. A new flavone C-glycoside was isolated and its structure was identified as 3'-methoxyl-luteolin 6-C-beta-D-galactopyranosiduronic acid (1 --> 2) -alpha-L-arabinopyranoside (1).
Antiviral Agents ; chemistry ; isolation & purification ; Flavones ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Hydrolysis ; Plant Leaves ; chemistry ; Poaceae ; chemistry

OBJECTIVETo compare the gene expression profiles of nasopharyngeal carcinoma (NPC) cell line 5-8F-EGFP and the liver metastatic 5-8F-H3B-EGFP cells.

METHODSThe fluorescence-labeled cDNA were prepared separately from the total RNA extracted from the two cell lines and hybridized with Human_U133A2.0 Genechip (Affymetrix, USA) containing approximately 18 400 known gene. The gene expression profiles were analyzed with special software and cluster analysis.

RESULTSA total of 3767 genes were identified to have significant differential expressions between these two cell lines (P<0.05), among which 281 genes showed twofold or higher differential expressions. Using MILANO software, we found 16 genes with probable close relation with liver metastasis of NPC.

CONCLUSIONThe 16 genes differentially expressed between the two cell lines can be of importance in the investigation of the molecular mechanism of NPC liver metastasis and identification of molecular markers for prognostic evaluation.

Carcinoma ; Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; genetics ; secondary ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Neoplasm Metastasis ; Oligonucleotide Array Sequence Analysis ; Transcriptome

Objective To explore the effect of Melatonin(MT) on CD4+CD25+ regulatory T cell (CD4+CD25+Tr)and airway inflammation in asthmatic rat.Methods Thirty-two SD rats were randomly divided into 4 groups,8 rats in each group.Asthmatic group:rats were immunized on day 1 and 7 by intraperitoneal inject of mixture of ovalbumin(OVA) and aluminumhydroxide.From day 14,the animals were allenged with aerosolized OVA for 20 min per day for 7 consecutive days.MT group:OVA-sensitized rats were injected intraperitoneally with 0.1 mg/kg MT 30 min before each OVA challenge.Dexamethasone group:OVA-sensitized rats were injected intraperitoneally with 0.5 mg/kg Dexamethasone 30 min before each OVA challenge.Control group:OVA for inhalation and MT for intraperitoneal injection was replaced with saline.After the last challenge,peripheral blood was stained to count the percentage of eosinophil(EOS).Then the rats were lavaged and total leukocytes counts in bronchoalveolar lavage fluid(BALF) were performed after staining with Wright-Giemsa staining.The EOS counts around the airway was counted after the histological section of lung staining with hematoxylin and eosin staining.The serum level of immunoglobulin E(IgE) was detected by immunoenhancement.The change of CD4+CD25+Tr was assessed with flow cytometry.SPSS 10.0 software was applied to analyze data. Results In asthmatic rats,the CD4+CD25+ Tr/ CD4+T cells ratio had significant negative relationship with the EOS counts around the airway and the total leukocytes counts in BALF (r=-0.73 P0.05).There was a significant decrease in the percentage of the eosinophils in peripheral blood,the eosinophil counts around the airway,the total leukocytes counts in BALF and the serum level of IgE in MT group compared with asthmatic group (Pa

OBJECTIVETo study the molecular mechanism underlying multidrug resistance (MDR) and identify unknown genes that might be involved in drug resistance development in K562/A02 cells.

METHODSK562/A02 was induced by gradually increasing the ADM concentration in culture medium of K562 cells, the differential expression of associated genes between K562 and K562/A02 was determined with cDNA microarray. Overexpression of neurofilament protein NF-H gene in K562/A02 cells was confirmed with RT-PCR and immunocytochemistry. Anti-sense oligodeoxynucleotides were transfected into K562/A02 cells by lipofectamine in order to further analyze the role of NF-H in drug resistance.

RESULTSComparing with the expression profiles, we found upregulation of 5 transcripts and downregulation of 7 transcripts in response to MDR of K562/A02 cells. The overexpression of NF-H, one of the 5 upregulated genes, was confirmed. After being treated with antisense oligodeoxynucleotides of NF-H and mdr1, the cellular adriamycin concentration increased significantly, but antisense NF-H alone did not have significant effect on drug resistance phenotype.

CONCLUSIONThe development of MDR in K562/A02 cells is multifactorial. NF-H may be involved in the drug resistance of K562/A02, which may provide a new marker of diagnosis and a new target of therapy.

Doxorubicin ; metabolism ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Genes, MDR ; Humans ; K562 Cells ; Neurofilament Proteins ; biosynthesis ; genetics ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; biosynthesis ; genetics ; Up-Regulation

AIMTo observe the expression of Presenilin-1 (PS-1) and production of amyloid beta-protein (Abeta) in hippocampus of female senile rats and to investigate the effect of vitamin E(VE) on preventing Alzheimer's disease after menopause.

METHODSThe animal model was established using female senile rats. Experimental groups (n=8) were respectively given different doses of VE(5 mg/kg, 15 mg/kg, 60 mg/kg) per day. The expression of PS-1 in hippocampus was detected by immunohistochemistry, the level of Abeta in hippocampus was measured by Radioimmunoassay, and neuronal ultrastructure in hippocampal DG area was observed using transmission electron microscope.

RESULTSThe expression of PS-1 in rat hippocampus of senile control group was stronger than that of adult control group. PS-1 expressed weakly in three medication groups along with augmentation of dosage. The levels of Abeta were found to correlate statistically with the expression of PS-1. The content of Abeta in VE groups was significantly decreased compared to that in senile control group (P < 0.01). There were some changes in the neuronal ultrastructure of senile rats. Neurons were gradually recovered in VE groups.

CONCLUSIONVE may depress the production of Abeta by regulating the expression of PS-1, reducing neuronal injuries. VE may play a role in neuronal protection.

Aging ; Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Female ; Hippocampus ; drug effects ; metabolism ; Presenilin-1 ; metabolism ; Rats ; Rats, Wistar ; Vitamin E ; pharmacology

NDRG2, one of the new N-Myc downstream-regulated gene (NDRG) gene families, is believed to be involved in cell growth event. However, the exact function is still unknown. Identification of the tissue or cell types expressing this gene in vivo will provide clues in clarifying its physiological roles. Using RT-PCR and Western blot, we analyzed the expression level of NDRG2 mRNA and protein in human fetal tissues from different gestational ages. The anti-NDRG2 monoclonal antibody, which has been proved to react specifically with NDRG2 protein, was further used to analyze the cellular location of NDRG2 protein in various human fetal tissues by immunohistochemistry. We found that NDRG2 expression was developmentally dynamic, being generally lower in the early stages of development and markedly increasing during the later stages. NDRG2 mRNA and protein distribution were generally consistent in heart and lung. One of the differences was that NDRG2 protein appeared later than mRNA in kidney. Another unmatched expression was found in liver. NDRG2 mRNA appeared later than protein in liver. In human fetal tissues at sixteen and twenty-eight weeks of gestation, NDRG2 protein immunoreactions could be seen in epithelium of small intestine, epithelium of large intestine, superficial layer of epidermis, whisker follicles, epithelium of small bronchus, hepatocytes, cardiac myocytes, thymus corpuscles and epithelium of renal tubule, and the immunoreactions in those tissues from twenty-eight weeks of gestation was stronger than that from sixteen weeks of gestation. In the present study, we demonstrate the expression pattern and cellular location of NDRG2 protein in a large set of human fetal tissues. This is the first demonstration of NDRG2 protein expression in human fetal tissues. Taken together, the results suggest that NDRG2 protein found in a variety of tissues is not a tissue-specific protein, and may play important roles in histogenesis and organogenesis.
Fetus ; Gene Expression Regulation, Developmental ; Humans ; RNA, Messenger ; genetics ; metabolism ; Tissue Distribution ; Tumor Suppressor Proteins ; genetics ; metabolism

In order to provide a new method for the identification of Placenta hominis, the COI barcode has been employed to identify the P. hominis medicinal materials and its adulterants. Genomic DNA was extracted from the experimental samples. The COI sequences were amplified and sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. NJ tree was constructed by MEGA6.0 software. COI sequences can be successfully obtained from all experimental samples. The intra-specific variation and inter-specific divergence were calculated. The average intra-specific K2P distance of P. hominis was 0.001 and the maximum intra-specific distance was 0.008. The cluster dendrogram constructed can be seen that the same genus is together, and distinguished from its adulterants. It is concluded that P. hominis and its adulterants can be correctly identified by DNA barcoding method.
Animals ; Cattle ; DNA Barcoding, Taxonomic ; methods ; Drug Contamination ; prevention & control ; Electron Transport Complex IV ; genetics ; Female ; Humans ; Medicine, Chinese Traditional ; Molecular Sequence Data ; Phylogeny ; Placenta ; chemistry ; enzymology ; Pregnancy ; Quality Control ; Sheep ; Swine

OBJECTIVESTo investigate the effect of gamma-aminobutyric acid (GABA) on the sperm acrosin activity in normal men and positive antisperm antibody (AsAb) men.

METHODSSperm acrosin activity was detected by BAEE/ADH method.

RESULTSGABA could increase the sperm acrosin activity in normal and AsAb positive patients (P < 0.01). The results also indicated that GABA significantly increased Na(+)-K(+)-ATPase activity (P < 0.01), Ca(2+)-ATPase activity (P < 0.05) and SOD activity (P < 0.01) of sperm.

CONCLUSIONSThe results demonstrated that GABA could influence the sperm acrosin activity.

Acrosin ; metabolism ; Humans ; Male ; Spermatozoa ; drug effects ; immunology ; metabolism ; gamma-Aminobutyric Acid ; pharmacology