3.Preparation of cell penetrating peptide TAT and cleavable PEGco-modified liposomes loaded with paclitaxel and its in vitro apoptosis assay.
Han FU ; Guan-Lian HU ; Qin HE
Acta Pharmaceutica Sinica 2014;49(7):1054-1061
The preparation method, serum stability, efficiency of cellular uptake and apoptosis induction of the cell penetrating peptide TAT and cleavable PEG co-modified liposomes loaded with paclitaxel (C-TAT-Lipo) were investigated. The best preparation procedure was performed by orthogonal test based on single factor screening method. First, the paclitaxel (PTX)-loaded liposomes were prepared by filming-rehydration method, evaluated with entrapment efficiency and polydispersity index. The morphology of C-TAT-Lipo was characterized by transmission electron microscopy. Turbidity variations were monitored in the presence of fetal bovine serum (FBS) to evaluate the serum stability of the liposomes developed here. Next, the efficiency of cellular uptake of different Rho-PE-labeled liposomes on B16F1 cells in vitro was evaluated by confocal laser scanning microscopy (CLSM) and flow cytometry. The quantitative analysis of apoptosis induced by different PTX-loaded liposomes was performed by Annexin V-FITC/PI double staining. The optimal formulation was as follows: Chol : lipid: 1 : 8 (molar ratio); drug : lipid: 1 : 40 (mass ratio); lipid concentration: 3 mmol x L(-1); temperature of hydration: 25 degrees C. The mean size and polydispersity index of C-TAT-Lipo were about (97.97 +/- 3.68) nm and 0.196 +/- 0.037, the zeta potential was (-0.89 +/- 0.45) mV, the entrapment efficiency of paclitaxel was (90.16 +/- 1.53)%. The particle sizes did not exhibit significant variations in 50% FBS over 24 h at 37 degrees C. The efficiency of cellular uptake of the C-TAT-Lipo increased 1.40 fold following the cleavage of PEG. Apoptosis analysis showed 59.3% increase of the apoptosis and necrosis profile of C-TAT-Lipo after the detachment of PEG shells, which was markedly higher than that of N-TAT-LP with or without glutathione and SL, respectively. The results indicate that the C-TAT-Lipo is successfully prepared by filming-rehydration method and shows significant antitumor activities.
Animals
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Annexin A5
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Apoptosis
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Cell Line, Tumor
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Cell-Penetrating Peptides
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pharmacology
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Fluorescein-5-isothiocyanate
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analogs & derivatives
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Liposomes
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chemistry
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Melanoma, Experimental
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Mice
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Microscopy, Confocal
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Paclitaxel
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pharmacology
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Particle Size
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Polyethylene Glycols
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chemistry
4.Effect of ferulic acid on cholesterol efflux in macrophage foam cell formation and potential mechanism.
China Journal of Chinese Materia Medica 2015;40(3):533-537
The formation of macrophage-derived foam cells is a typical feature of atherosclerosis (AS). Reverse cholesterol efflux (RCT) is one of important factors for the formation of macrophage foam cells. In this study, macrophage form cells were induced by oxidized low density lipoprotein (ox-LDL) and then treated with different concentrations of ferulic acid, so as to observe the effect of ferulic acid on the intracellular lipid metabolism in the ox-LDL-induced macrophage foam cell formation, the cholesterol efflux and the mRNA expression and protein levels of ATP binding cassette transporter A1 (ABCA1) and ATP binding cassette transporter G1 (ABCG1) that mediate cholesterol efflux, and discuss the potential mechanism of ferulic acid in resisting AS. According to the findings, compared with the control group, the ox-LDL-treated group showed significant increase in intracellular lipid content, especially for the cholesterol content; whereas the intracellular lipid accumulation markedly decreased, after the treatment with ferulic acid. The data also demonstrated that the mRNA and protein expressions of ABCA1 and ABCG1 significantly increased after macrophage foam cells were treated with different concentrations of ferulic acid. In summary, ferulic acid may show the anti-atherosclerosis effect by increasing the surface ABCA1 and ABCG1 expressions of macrophage form cells and promoting cholesterol efflux.
ATP Binding Cassette Transporter 1
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analysis
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genetics
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ATP Binding Cassette Transporter, Sub-Family G, Member 1
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ATP-Binding Cassette Transporters
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analysis
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genetics
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Animals
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Cells, Cultured
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Cholesterol
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metabolism
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Coumaric Acids
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pharmacology
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Foam Cells
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drug effects
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metabolism
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Lipoproteins
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analysis
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genetics
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Mice
5.Expression of Hypoxia Inducible Factor-1? in Vertebra Development of Mouse
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(05):-
Objective To investigate the expression pattern of hypoxia inducible factor-1?(HIF-1?) in fetal vertebra development of the mouse. Methods The development of mouse fetal vertebra was observed dynamically,and the expression of HIF-1? mRNA at various stages was also detected by reverse transcription-polymerase chain reaction. Results The cartilaginous spine column began to form at E13.5.The primary ossification center was observed at E15.5,then the osteogenesis expanded and extended to both sides.HIF-1? mRNA began to express at E13.5,and more significantly at E14.5(P
6.Awareness rate of child health management in parents of children aged 0 - 36 months
Xiaobing FU ; Jie LIAN ; Ping SHI
Chinese Journal of General Practitioners 2012;(7):519-521
A questionnaire survey on awareness of child health management was conducted in 238 family members ( 236 were parents) of children aged 0 - 36 months; among whom 92 of children aged 0 to 6 months,85 of children aged 7 to 12 months,33 of children aged 13 to 18 months,16 of children aged 19 to 24 months and 12 of children aged 24 to 36 months.52.1% of the surveyees had college education level.The awareness about medical examination in last half year in female subjects was higher than males ( P < 0.05).There were no differences in other health management knowledge.The results showed that the average times of medical examination was 2.8 ± 1.9 in last half year; the age for supplement of meat in food was 7.4 ± 2.4 mouths; supplement of meat,animal liver and eggs was more than 7 time a week.The subjects with college education or above paid more attention to the children's growth,risk for rickets and anemia than other peoples.Pure breast feeding accounted for only 37.0% (88/238).The study indicates that strengthening the knowledge and awareness is necessary to improve the children health management.
7.Experimental study on electroencephalographic characteristic in kainic acid induced temporal lobe epilepsy model established with stereotactic technique in rats
Ruobing QIAN ; Xianming FU ; Lian YU
Journal of Clinical Neurology 1992;0(01):-
Objective To explore the method for establish the temporal lobe epilepsy model in SD rats with stereotactic technique and to study the characteristic of electroencephalography (EEG) after epileptic seizure. Methods Kainic acid was injected into right hippocampus via stereotactic technique in 20 rats, and the behavior of rat was recorded by a video camera. The electrical activity changes of brain were recorded by EEG in bilateral hippocampus, amygdala and frontal lobe. Results According to Racine's scale, 6 rats were typeⅤ,10 rats were type Ⅳ,3 rats were type Ⅲ,1 rat was type Ⅱ. The spike wave was originated from hippocampus and spread to amygdala and frontal lobe. Pathologic changes showed that prolonged seizures induced pyramidal cell loss and death in hippocampus. Conclusions The method to establish kainic acid induced temporal lobe epilepsy model with stereotactic technique was credible, economical and useful. The spike wave originated from hippocampus was the basic feature of EEG of temporal lobe epilepsy model in rats.
10.Effects of Three Herb Extracts on Cell Proliferation and Collagen Synthesis in Cultured Human Dermal Fi-broblasts
Lian XU ; Xudong FU ; Rui XIONG ; Mengying LUO
China Pharmacist 2016;19(3):448-452
Objective:To investigate the effects of three herb extracts on cell proliferation and collagen synthesis in cultured hu-man dermal fibroblasts. Methods:The human dermal fibroblasts were isolated and cultured with tissue explant culture technique. Im-munohistochemical staining was performed to observe vimentin in fibroblasts. The proliferation of fibroblasts was determined by MTT as-say. Hydroxyproline in the cultural medium was determined by a digestion method. Results:The cells were typical fibroblasts, and the immunohistochemistry staining of vimentin in the cells was positive. The three herb extracts could promote the proliferation of the fibro-blasts at appropriate concentrations in a time-dependent manner. After the 72-hour culture in the medium, 0. 312 5-5. 000 0 mg·ml-1 Aloe vera gel extract, 0. 156 3-2. 500 0 mg·ml-1 Bletilla striata extract and 0. 075 0 mg·ml-1 salvianolate showed significant effects on the proliferation of fibroblasts (P<0. 05 or P<0. 01 vs the blank group). Compared with that in the blank group, the amount of hydroxyproline in the three herb extracts cultural medium was also increased (P<0. 05 or P<0. 01). The effect of salvianolate on the cell proliferation was closely related with time and concentration, and the high concentration group could inhibit the growth of fibroblasts as the extension of time. Conclusion:Aloe vera gel extract, bletilla striata extract and salvianolate can promote the proliferation of fi-broblasts and the production of hydroxyproline, which may contribute to the wound healing.