Objective To investigate the depressing effect of antigene peptide nucleic acid (AGPNA)on the k-ras gene expression of human pancreatic cancer Patu8988 cells, the inducing apoptosis effect on Patu8988 cells, and the biodistribution characteristics in nude mice bearing xenografts using 188Re-k-ras-AGPNA.Methods The expression level of k-ras mRNA and the expression ratio of k-ras protein in Patu8988 cells transfected with AGPNA was measured by RT-PCR and flow cytometry ,respectively. The degree of cellular apoptosis 3 to 5 d after treating Patu8988 cells with 188Re-k-ras-AGPNA or 188ReO4- was determined by flow cytometry. For biodistribution study, 58 nude mice bearing Patu8988 cell xenografts were divided into two groups: intratumoral injection of 188Re-k-ras-AGPNA (Group A) and 188ReO4- (Group B). At different time points, the mice were sacrificed and organs of interest were excised, weighted and counted by a gamma counter. The organ uptake was calculated as a % ID/g and the absorbed doses of organs were calculated. One-way analysis of variance was used. Results After transfected with 1 nmol/ml AGPNA, the k-ras mRNA gray scale ratio and the expression ratio of k-ras protein were 1.00 ± 0.39 and (15.05 ± 5.07)%, respectively. They were significantly lower than those of the control group with 1.86 ± 0.07 and (24. 38 ± 5.40) % (F = 2. 545, 5. 327, P<0. 05). At 4 and 5 d after treatment in Group A, float cells' apoptosis ratios were (26.30 ± 7.45) % and (27.90 ± 10. 38) %, respectively. Tumors were the major distribution site in Group A with uptake of (37.47 ±21.31), (35.96 ±7.80) and (15.46 ±4.93) %lD/g at 1 h, 1 d and 7 d after intra-tumor injection, respectively. The absorbed dose of tumor was 15 569 mGy/MBq. Condusions Transfection with k-ras-AGPNA on Patu8988 cells may inhibit k-ras expression at mRNA and protein expression level, and 188Re-k-ras-AGPNA can induce apoptosis of Patu8988 cells.Tumor is the major distribution site in nude mice bearing human pancreatic cancer xenografts after intratumoral injection of 188Re-k-ras-AGPNA.

Objective To evaluate the killing effect and the uptake of 125I-UdR on human lymphoma Raji and Daudi cell lines. Methods The amount of 125I-UdR in the cells and cell nuclei were determined after incubation of different time in RPMI 1640 culturing medium containing different concentrations of 125I-UdR. The killing effects of 125I-UdR on Raji and Daudi cell lines were estimated through MTT assay and cell cycle was analyzed by propidium iodide (PI) staining. Results The amounts of 125I-UdR in Raji and Dandi cells and cell nuclei were much higher than that of Na125I(P < 0.05). The amounts of 125l-UdR in Raji and Daudi cells were 14414±95 and (6916± 53.69) Bq/106 cell when the concentration was 100 kBq/ml. The amounts of Na125I were 68± 3.8 and (324±32.8) Bq/106 cell. The uptake of 125I-UdR in Raji and Daudi cells and cell nuclei increased with the 125I-UdR concentration and incubated time. The cell surviving fractions of 125I-UdR groups was much lower than that of Na125I groups (P < 0.05). When the concentration was 500 kBq/ml and incubated time was 48 hours, the Raji and Dandi cell surviving fractions of125I-UdR groups were (19.78 ± 1.39)% and (43.17 ± 2.69) % ;those of Na125I groups were (79.10 ± 1.79) % and (80.36 ± 6.12) %. The surviving fractions of 125I-UdR groups reduced with the 125I-UdR concentration. Conclusions 125I-UdR can be specially ingested by Raji and Daudi cells and incorporated into DNA, then the cells will be killed. The uptake of 125I-UdR is dose and time dependent.

Aim To investigate the lipid-lowering mechanism of Chinese Medicine curcumin.A sterol regulatory report system of LDLR gene expression in HEK-293 cell line was established.Method A green fluorescent protein(GFP) gene was constructed downstream of the sterol regulatory element-1(SRE-1) and the constructed plasmid was introduced into HEK-293 cell line.This SRE-GFP transfected 293 cells were treated with different concentration of curcumin for 2 days.The expression of GFP in the cells was measured by a fluorescence microscope and Flow Cytometry.Results The expression of GFP in SRE-GFP transfected 293 cells was obviously induced by curcumin.Conclusion Curcumin may positively affect the expression of LDLR gene by its influence on SRE-1.

Aim To investigate and evaluate the effect of ligustrazine on the expression of scavenger receptor type AI, the human ?-interferon activation site (GAS) elements regulatory systems of SR-AI expression wasfirst established in the Xenopus oocytes. Methods A plasmid, in which the human SR-AI gene was constructed downstream from the GAS elements, was microinjected into the nucleus of Xenopus oocytes. The oocytes then were cultured in the smooth muscle cell conditioned medium (SCM), In which the cell were cultured with the treatment of oxLDL for 2 days. In the other groups, the oocytes were treated with SCM containing the nature medicines ligustrazine, and the positive control drug ?-interferon respectively. Results The results demonstrated that SCM could upregulate the expression of SR-AI in this system, this action was suppressed by ligustrazine and ?-interferon. Conclusion Similar to the action of ?-interferon, ligustrazine can prevent the over-expression of SR-A induced by oxLDL, via their effect on the GAS elements regulatory pathway.

Objective To compare the incorporation method of 3H-TdR and 125Ⅰ-UdR on determining the proliferation effect of lymphocytes. Methods The proliferation effects of lymphocyte and Daudi lymphoma cells were estimated by 3H-TdR and 125Ⅰ-UdR incorporation. Results The incorporating fraction of 3H-TdR and 125Ⅰ-UdR into lymphocyte was 20.95% ± 1.06% and 1.00% ±0.04%,respectively, and the incorporating fraction for the lymphoma cells was 29. 94% ± 4. 10% and 6. 02% ±0. 73% ,respectively. The incorporation fractions of 3H-TdR into lymphocyte and lymphoma cells were much higher than those of 125Ⅰ-UdR, but the incorporating fractions of 3H-TdR or 125Ⅰ-UdR into the lymphoma cells were much higher than those of lymphocytes. Conclusions For lymphocytes, 125Ⅰ-UdR cannot substitute 3H-TdR as a tracer agent. But for lymphoma cells, whether 125Ⅰ-UdR could be replace 3H-TdR or not needs further research.

Animals ; Antibodies, Monoclonal ; pharmacokinetics ; Iodine Radioisotopes ; pharmacokinetics ; Leukocyte Common Antigens ; immunology ; Male ; Mice ; Radiation Monitoring ; methods ; Rats ; Tissue Distribution

BACKGROUNDPeroxisome proliferator-activated receptor-gamma (PPARgamma) is a kind of ligand-activated transcription factors binding to peroxisome proliferator response element (PPRE), a specific recognition site. It is thought to play a critical role in glucose and lipid metabolism and in inflammation control. The aim of this study was to establish a new cellular model for the quick screening of lipid-lowering drugs, which may be effective as PPAR-gamma ligands on the PPRE-mediated pathway regulatory system.

METHODSTwo plasmids were constructed: pXOE-PPARgamma, in which the human PPARgamma gene was in the downstream of TFIIIA gene promoter, and pLXRN-PPRE-d2EGFP, in which the enhanced green fluorescent protein (EGFP) gene was subcloned into PPRE. The xenopus oocytes were injected with these two plasmids, and consequently treated with prostaglandin E1, pioglitazone, and different kinds of lipid-lowering drugs. After 3 days, the oocytes were observed under a fluorescence microscope. To confirm the drug action,we injected pXOE-PPARgamma plasmid into the oocytes, which then treated with prostaglandin E1 and Hawthorn flavonoids. The mass of expressed lipoprotein lipase (LPL) in the cells was determined by enzyme labeling linked immunosorbent assay (ELISA).

RESULTSThe expression of EGFP was only induced by prostagalandin E1, pioglitazone, Hawthorn flavonoids. A concentration-response relationship was seen between expressed EGFP and Hawthorn flavonoids. The levels of LPL in both Hawthorn flavonoids groups and PPARgamma ligand prostagalandin E1 group injected with pXOE-PPARgamma plasmid increased significantly (< 0.001) compared with controls, and a concentration-response relationship was observed between LPL mass and Hawthorn flavonoids.

CONCLUSIONSIt is possible to establish a PPRE regulatory EGFP reporter system in xenopus oocytes to monitor the activity of PPARgamma ligand. Hawthorn flavonoids can increase the expression of gene downsteam of PPRE by effect on the PPRE pathway regulatory system.

Alprostadil ; pharmacology ; Animals ; Crataegus ; Female ; Hypolipidemic Agents ; pharmacology ; Lipoprotein Lipase ; biosynthesis ; Medicine, Chinese Traditional ; Oocytes ; metabolism ; PPAR gamma ; physiology ; Peroxisome Proliferators ; pharmacology ; Plasmids ; Response Elements ; physiology ; Xenopus

OBJECTIVETo investigate the molecular mechanism of curcumin in reducing blood lipids by establishing gene expression system of human low density lipoprotein receptors (LDL-R) in Xenopus Laevis oocytes (XLO).

METHODSThe expression of LDL-R on cytomembrane was determined using immuno-fluorescent, ligand-fluorescent and immune colloidal gold techniques after human LDL-R containing p3.7 LDL plasmid was led into nucleus. And the expression of LDL-R gene in XLO was quantitatively determined by ELISA after being interfered with different concentrations of curcumin.

RESULTSThe human LDL-R gene could be expressed on XLO, which could be significantly enhanced by curcumin in a dose-dependent manner. Conclusion One of the paths of curcumin in reducing blood lipids and anti-atherosclerosis was improving LDL-R gene expression and increasing the LDL-cholesterol absorption of cells.

Animals ; Cells, Cultured ; Curcumin ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Gene Expression Regulation ; Humans ; Microinjections ; Oocytes ; cytology ; metabolism ; Receptors, LDL ; biosynthesis ; genetics ; Xenopus laevis

Hyperlipidemia (HLP) is the No.1 risk factor for patients with atherosclerosis (AS) and is directly related to the occurrence of coronary artery disease (CAD) and cerebrovascular disease. Therefore, prevention and treatment of AS is of great importance and of practical significance in controlling the incidence and mortality of CAD. With its peculiar syndrome-dependent therapy, traditional Chinese medicine (TCM) has accumulated abundant practical experiences in this field and good clinical effects have been achieved. Chinese herbal medicine, with its particularly unique advantages and high potentials yet to be tapped, displays its huge strength in HLP prevention and treatment. The progress of studies concerning prevention and treatment of HLP by Chinese herbal medicines, in the form of monomers or compound recipes, is reviewed in this paper.
Cholesterol ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Hyperlipidemias ; drug therapy ; Lipid Metabolism ; Lipid Peroxidation ; Receptors, LDL ; analysis

IL-8 was radiolabeled with 125I via Bolten-Hunter agent and the distribution of 125I-IL-8 in mice was determined in order to understand IL-8 behaviors in vivo. The percentage of 125I-IL-8 in blood, heart, liver, lung, kidneys, bone and spleen was obtained. The half-clearance of fast phase T1/2 alpha and slow phase T1/2 beta were 0.32 h and 8.01 h respectively. Most of 125I-IL-8 was excluded by kidneys.
Animals ; Interleukin-8 ; pharmacokinetics ; Iodine Radioisotopes ; Male ; Mice ; Recombinant Proteins ; pharmacokinetics ; Tissue Distribution