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

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 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

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 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

The present study was undertaken to investigate the effects of ceramide on progesterone production and apoptosis in rat luteal cells in vitro. Luteal cells were prepared from the ovaries of pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) primed female Wistar rats and incubated with cell permeable C2-ceramide. The effects of ceramide on progesterone production and apoptosis in luteal cells were assessed by radio immunoassay (RIA) and flow cytometry analysis. In addition, changes in nitric oxide synthase (NOS) activity and nitric oxide (NO) production by luteal cells treated with C2-ceramide were also evaluated. Ceramide was found to reduce hCG-stimulated progesterone production in a dose-dependent manner, whereas it had little effect on basal progesterone content. Spontaneous apoptosis in luteal cells was observed after a 12-hour incubation in vitro and 5 μmol/L of ceramide significantly increased the apoptotic rate (P<0.05). Enhanced apoptotic peak was seen in the histogram by flow cytometry. Moreover, 50 μmol/L of C2-ceramide significantly increased NOS activity (P<0.01) and NO production (P<0.001). It is suggested that ceramide may serve as an important signaling molecule mediating certain ovarian processes, such as luteal regression.

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

OBJECTIVETo explore the therapeutic mechanisms of interferon-beta (IFN-beta) and intravenous immunoglobulin (IVIG) for experimental peripheral neuropathy induced by Campilobacter jejuni (Cj) lipopolysaccharide (LPS).

METHODForty healthy Wistar rats weighing 205 - 230 g were divided into IFN-beta, IVIG, IFN-beta plus IVIG and control groups. After the immune neuropathy was induced in the rats by Cj LPS, IFN-beta (1.3 microg/kg) was given by subcutaneous injection to the rats every other day for 6 weeks; IVIG [400 mg/(kg x d)] was given to the rats for five days, every other week for two times and IFN-beta [1.3 microg/(kg x d)] and IVIG [400 mg/(kg x d)] were given to the rats on the same days. Meanwhile, the control group was given PBS. The sera were collected in the 2nd, 4th and 6th week after therapy, the titers of anti-GM(1) IgG, MMP-9 and TNF-alpha in sera of immunized rats were measured by ELISA; histological study of sciatic nerve was performed and IgG on sciatic nerve was detected by immunohistochemistry in the 6th week.

RESULTS(1) There were no significant differences in titers of anti-GM(1) IgG, MMP-9 and TNF-alpha among the 3 therapeutic groups and control group after therapy for 2 weeks (P > 0.05). (2) The titers of anti- GM(1) IgG, MMP-9 or TNF-alpha in the control group were much higher than those of the IFN-beta group, the IVIG group or the IFN-beta and IVIG group after therapy for 4 weeks (P > 0.01) and there were no significant differences in titers of antibody among the 3 therapeutic groups (P > 0.05); the titers of MMP-9 or TNF-alpha in the IFN-beta and IVIG group were lower than those of the IFN-beta group or the IVIG group (P < 0.05). (3) The titers of anti-GM(1) IgG, MMP-9 or TNF-alpha in the control group were much higher than those of the IFN-beta group, the IVIG group or the IFN-beta with IVIG group after therapy for 6 weeks (P > 0.01); the IFN-beta with IVIG group had much lower levels of all indexes than the IFN-beta group or the IVIG group (P < 0.01).

CONCLUSIONIFN-beta and IVIG showed therapeutic effects on immune peripheral neuropathy through inhibiting the humoral and cellular immunity simultaneously in the peripheral neuropathy induced by CJ LPS, treatment with combined IFN-beta and IVIG was more effective.

Animals ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulins, Intravenous ; therapeutic use ; Immunotherapy ; Interferon Type I ; therapeutic use ; Interferon-beta ; immunology ; therapeutic use ; Lipopolysaccharides ; pharmacology ; Peripheral Nervous System Diseases ; therapy ; Rats ; Rats, Wistar ; Recombinant Proteins ; Sciatic Nerve ; drug effects ; Tumor Necrosis Factor-alpha ; immunology

AIMTo observe the effect of vitamin E (VE) on ovarian apoptosis-related protein Bcl-2 and Bax and its impact on antioxidant capacity in aged female rats and to study the senility-delaying effect and mechanism of VE on ovary.

METHODSNatural aging female rats were given different doses of exogenous VE. Then apoptosis regulatory protein Bcl-2, Bax expression in ovarian grandlose cells were detected by using immunohistochemical methods and Western blot. The contents of serum total superoxide dismutase (SOD) activity and malondialdehyde (MDA) were detected by using biochemical methods.

RESULTSContrasted with adult control group, the level of Bcl-2 expression in Senile control group was lower and the level of Bax expression was higher (P < 0.01), Serum SOD activity decreased and the level of MDA significantly increased (P < 0.01). Contrasted with senile control group, the level of Bcl-2 expression increased in VE group, the level of Bax expression decreased (P < 0.05), the level of MDA expression significantly decreased (P < 0.01).

CONCLUSIONVE can regulate apoptosis-related protein Bcl-2, Bax expression and confront free radical damage which contribute to a protective effect for ovarian grandiose cells.

Aging ; Animals ; Antioxidants ; pharmacology ; Apoptosis ; drug effects ; Female ; Granulosa Cells ; cytology ; Ovary ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; Vitamin E ; pharmacology ; bcl-2-Associated X Protein ; metabolism

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