No abstract available.
Crohn Disease/*drug therapy ; Female ; Humans ; Immunosuppressive Agents/*administration & dosage ; Male ; Oligonucleotides/*administration & dosage ; Smad7 Protein/*antagonists & inhibitors

AIMTo investigate the possibility of polymethacrylate nanoparticles (NP) for antisense oligodeoxynucleotides delivery system.

METHODSThe nanoparticles were prepared by evaporating ethenol solution containing Eudragit RL100 or RS100, and then mixtured with oligonucleotides. The morphology and size were investigated by a transmission electron microscope and Mastersizer particle characterization systems, and the cytotoxicity was evaluated by Trypan Blue staining and hemolysis test. The flow cytometer was used to determine the uptake of fluorescence-labelled oligodeoxynucleotides.

RESULTSThe morphology of nanoparticles showed spherical and orderly, the average diameter was about 127 nm, and almost the antisense oligodeoxynucleotides (ODN) were loaded when NP: ODN was 6.6. The uptake of ODN was significantly increased when loaded by nanoparticles, which well depended on the nanoparticles concentration. Meanwhile, slightly cytotoxicity was observed when high dose of nanoparticles was used.

CONCLUSIONThe polymethacrylate nanoparticles appeared to be a promising vehicle for gene delivery.

Acrylic Resins ; chemistry ; toxicity ; Animals ; Drug Carriers ; Drug Delivery Systems ; Hemolysis ; drug effects ; Nanotechnology ; Oligonucleotides, Antisense ; administration & dosage ; Particle Size ; Technology, Pharmaceutical ; methods

AIMTo prepare the liposomes which protect antisense oligodeoxynucleotides (ASON) against nuclease degradation and delivery ASON into cytoplasmic efficiently.

METHODSA cationic derivative of cholesterol, 3 beta-[N-(N',N'-dimethylaminoethan)-carbamoyl] cholesterol (DC-Chol) was synthesized and used to prepare cationic liposome. The characteristics of liposomes/ASON complexes including size, drug loaded efficiency and structure were investigated. Cellular uptake of fluorescence labled ASON (FAM-ASON) under different condition was determined by flow cytometric analysis. Denatured polyacryamide gel electrophoresis (DPGE) was used to analyze the role of liposomes in protecting ASON.

RESULTSThe mean values of preliposomes and liposomes/ASON complexes size were 185.7 and 228.2 nm, respectively. Cationic liposomes showed a high adsorption capacity for ASON. When the +/- charge ratio exceeded 2:1, more than 90% of the ASON was loaded into liposomes. Agarose gel electrophoresis showed three different existence of ASON in liposomes formulation: free, absorbed and encapsulated types. Concerning cellular uptake, DC-Chol liposomes indicated high efficient effect of increasing cellular uptake of ASON. Compared with free ASON, the total fluorescence intensity in cytoplasma was significantly enhanced. The level of increasing was largely depended on +/- charge ratio. The cellular uptake of FAM-ASON decreased in the presence of serum. The cellular total fluorescence intensity in 10% and 30% fetal bovine serum of cultured medium were only 22.3% and 15.5% as that of serum-free media, respectively. DPGE confirmed that free ASON was rapidly degraded by DNase I while ASON encapsulated into liposomes was efficiently protected.

CONCLUSIONThe cationic DC-Chol liposomes are shown to be promising carriers to deliver ASON into cytoplasma.

Cholesterol ; analogs & derivatives ; pharmacology ; Drug Carriers ; Drug Delivery Systems ; HeLa Cells ; Humans ; Liposomes ; pharmacology ; Multiple Myeloma ; pathology ; Oligonucleotides, Antisense ; administration & dosage ; blood ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo evaluate the antitumor efficacy of proliferating cell nuclear antigen antisense oligonucleotide (PCNA-ASO) in combination with recombinant adenovirus p53 (Ad-p53) against bladder cancer EJ and BIU-87 cells in vitro and in vivo.

METHODSCells were transfected with Ad-p53 (100 MOI), and PCNA-ASO (1.6 micro mol/L) was then introduced into the cells using a cationic lipid (lipofectamine, 20 micro l/ml). In vitro and in vivo antitumor effects of combining PCNA-ASO with Ad-p53 were measured using the MTT assay, flow cytometry, clone formation, and a nude mice model.

RESULTSThe combination of PCNA-ASO and Ad-p53 inhibited cell viability in both the EJ (89.3%) and BIU-87 (78.6%) cell lines. The ability of the cells to form foci was also reduced by 74.8% in EJ cells and by 67.5% in BIU-87 cells (P < 0.01). A significant decrease of cells in the S phase (11.4% in EJ cells, 14.6% in BIU-87 cells) and a significant increase of cells in G1 phase (62.2% in EJ, 56.8% in BIU-87) were noted. The mean tumor volume after 7 days of treatment with PCNA-ASO or Ad-p53 in combination decreased to 47.6% or 36.4% of the initial tumor size in the two cell lines respectively.

CONCLUSIONThese results indicate that combined PCNA-ASO and Ad-p53 in the treatment of bladder cancer with mutant p53 has important therapeutic potential, significantly suppressing the growth of human bladder cancer both in vitro and in vivo.

Adenoviridae ; Animals ; Genetic Therapy ; methods ; Genetic Vectors ; Humans ; Male ; Mice ; Mice, Nude ; Oligonucleotides, Antisense ; administration & dosage ; Proliferating Cell Nuclear Antigen ; administration & dosage ; Recombinant Proteins ; Transfection ; Tumor Cells, Cultured ; Tumor Suppressor Protein p53 ; administration & dosage ; Urinary Bladder Neoplasms ; therapy

OBJECTIVETo identify the effect of PNA CXCR3 on acute rejection of islet allograft.

METHODSThe mice islet transplant models were used. The mice were divided into three groups including saline group, PNA CXCR3 group and mismatch PNA group. In vitro the proliferation capability of T cell was assessed by proliferative responses. RT-PCR and western blot were used to detect the expression of mRNA and protein. Flow cytometry was applied to determine the expression level of CXCR3 in spleen CD3(+) T cells.

RESULTSCompared with saline [(6.72 +/- 1.48) d] and PNA mismatch-treated recipients [(6.54 +/- 0.86) d], PNA CXCR3-treated recipients demonstrated statistically significant prolongation [(9.70 +/- 1.57) d] in functional allograft survival. The CXCR3 mRNA expression level of PNA CXCR3 group (1.06 +/- 0.07) was significantly down-regulated compared with saline (1.98 +/- 0.22) and PNA mismatch (1.87 +/- 0.10) group at the 7th day after transplant. The date showed that CXCR3 protein and lymphocytes proliferation capability was significantly down-regulated in PNA CXCR3 group compared with saline and PNA mismatch group (P<0.01).

CONCLUSIONSThe present study indicates that PNA CXCR3 can inhibit T cell activating and prolonging the survival time of islet allograft and has a substantial therapeutic effect on inhibiting acute allograft rejection.

Animals ; Blotting, Western ; Diabetes Mellitus, Experimental ; surgery ; Graft Rejection ; genetics ; physiopathology ; Graft Survival ; drug effects ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Oligonucleotides, Antisense ; administration & dosage ; genetics ; Pancreas Transplantation ; methods ; Peptide Nucleic Acids ; administration & dosage ; genetics ; Random Allocation ; Receptors, CXCR3 ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; genetics ; physiology ; Transplantation, Homologous

OBJECTIVETo study the inhibitory effect of antisense oligonucleotides against telomerase RNA on the growth of human choriocarcinoma transplant in nude mice.

METHODSChoriocarcinoma xenografts were established by transplanting JAR cells subcutaneously to female nude mice, and were treated with high and low doses of antisense oligonucleotides. Control groups were treated with NS, random sequence and actinomycin D (Act-D). Tumor growth was monitored once every other day. Telomerase relative activity was assayed by TRAP-ELISA. Western blotting was used to detect expression of hTERT.

RESULTSLow and high doses antisense oligonucleotides, and Act-D inhibited tumor growth by 76.6%, 93.8% and 85.4% respectively, which were significantly different when compared with random sequence and NS groups. Expression of telomerase relative activity and hTERT were decreased as well. But the differences among the first three groups had no significance.

CONCLUSIONTelomerase RNA antisense oligonucleotide inhibits growth of human choriocarcinoma xenografts in nude mice. It may be a novel approach to the treatment of choriocarcinoma.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; Cell Line, Tumor ; Choriocarcinoma ; enzymology ; pathology ; DNA-Binding Proteins ; metabolism ; Dactinomycin ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; administration & dosage ; pharmacology ; Pregnancy ; Telomerase ; genetics ; metabolism ; Uterine Neoplasms ; enzymology ; pathology

OBJECTIVE: To characterize the antisense c-fos oligonucleotides that control the expression of immediate-early gene c-fos in retina in order to better understand the mechanism by which antisense c-fos oligonucleotides induced myopia. In this study the signal transduction in the pathway linking visual experience and the regulation of the eye's growth was investigated. METHODS: Thirty-one 3-week guinea pigs were assigned into 3 groups: antisense and sense c-fos oligonucleotides were intravitreally injected every 3 days to the eyes of the experimental guinea pigs at different concentrations; and saline vehicle to control guinea pigs in the same way. The refraction and axial length of the eyes were measured before and after the treatment, and the immediate-early gene c-fos expression in the retina was quantified by immunohistochemistry and RT-PCR. RESULTS: The moderate myopia was induced in high (1 nmol) and low (0.1 nmol) level of antisense c-fos oligonucleotide intravitreous injection (-5.425 D and -5.575 D, respectively) compared with the control ateral eyes. The refraction and axial length of the treated eyes increased, and the expression of immediate-early gene c-fos decreased significantly in the antisense c-fos oligonucleotides intravitreously injected eyes compared with the sense c-fos oligonucleotide intravitreously and saline vehicle injected eyes (P<0.01). The refraction and axial length were of no statistically significant differences among the sense c-fos oligonucleotides-treated eyes and saline-treated eyes and non-treated eyes (P>0.05). CONCLUSION The obvious myopia can be induced by antisense c-fos oligonucleotides in guinea pigs; antisense c-fos oligonucleotides inhibit c-fos expression in the retina. Immediate-early gene c-fos may be a potential factor in the prevention of myopia and plays an important role in the signal transduction of the retina.
Animals ; Genes, Immediate-Early ; genetics ; Guinea Pigs ; Immunohistochemistry ; Microinjections ; Myopia ; chemically induced ; genetics ; physiopathology ; Oligonucleotides, Antisense ; administration & dosage ; genetics ; toxicity ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Retina ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; physiology