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Objective:To investigate the preparation of paclitaxel-loaded nanoparticles targeting liver cancer stem cells and their effects on liver cancer HepG2 cells (CD133 positive subset accounting for 8%) and Huh-7 cells (CD133 positive subset accounting for 65%).Methods:Poly (lactic-co-glycolic acid)-loaded paclitaxel nanoparticles were prepared by using emulsification-solvent evaporation method. Paclitaxel-loaded nanoparticles decorated with anti-CD133 antibody, called targeted nanoparticles, were prepared by using 1- (3-dimethylaminopropyl)-3-ethylcarbodiimide/N-hydroxysuccinimide (EDC/NHS) cross-linking method. The manifestations and physicochemical characteristics of the nanoparticles including encapsulation efficiency, loading efficiency, particle size distribution, morphology and release in vitro were studied. Liver cancer Huh-7 and HepG2 cells accompanying with paclitaxel-loaded nanoparticles or targeted nanoparticles were cocultured. The uptake and accumulation of nanoparticles by liver cancer cells were analyzed by using flow cytometry, and positive cell proportion of CD133 was also detected. Cell survival was analyzed by using plate clonogenesis assay.Results:Scan electromicroscopy result showed particle size of targeted nanoparticles was (429.26±41.53) nm with zeta potential of -11.2 mV; targeted nanoparticles were possessed with spherical morphology and higher encapsulation efficiency [(87.53±5.90) %]. Flow cytometry showed that in Huh-7 cells at 37℃, the fluorescence intensity of targeted nanoparticles group (13 397±720) was higher than that of paclitaxel-loaded nanoparticles group (6 898±604), and the difference was statistically significant ( P < 0.05); there was no statistically difference in the fluorescence intensity of HepG2 cells in paclitaxel-loaded nanoparticles group and targeted nanoparticles group at 37 ℃ (7 899±343 vs. 8 432±516, P>0.05). CD133 positive cell proportion of Huh-7 cells in targeted nanoparticles group [(15.7±2.6)%] was lower than that in paclitaxel-loaded nanoparticles group [(54.9±7.4)%], and the difference was statistically significant ( t = 7.31, P = 0.008); there was no statistical difference of HepG2 cells between the two goups ( P > 0.05). Plate clonogenesis assay showed that the cell survival rate of Huh-7 cells in targeted nanoparticles group was lower than that in paclitaxel-loaded nanoparticles group at different time points, and the difference was statistically significant ( F = 5.56, P = 0.009); but there was no statistically significant difference in HepG2 cell survival rate between the two groups ( F = 1. 19, P = 0.142). Conclusion:Prepared nanoparticles targeting liver cancer stem cells have a good inhibitory effect on liver cancer Huh-7 cells.
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Objective@#To investigate the characterization of doxorubicin (DOX) coupling segmented copolymer nanomicelles with dual effects of passive and active targeting to liver cancer and its antineoplastic function in vitro.@*Methods@#DOX was covalently conjugated to a terminal hydroxyl group of poly lactic-co-glycolic acid-poly ethylene glycol (PLGA-PEG) diblock copolymer to form DOX-PLGA-PEG. The formation of amido bond was determined by using fourier transform infrared spectroscopy (FTIR) and magnetic resonance method. Amphiphatic diblock copolymer DOX-PLGA-PEG could self-assemble to form nanomicelles in an aqueous phase by dialysis method. The DOX-PLGA-PEG targeted micelles decorated with liver cancer HAb18 F (ab')2 specific antibody were prepared by using physical bonding method. The size and the scattering scope of nanomicelles was determined by using granulometer and dynamic light scattering (DLS). Micelle morphology was examined by using scanning electron microscopy (SEM). The drug loading rate and entrapment rate of DOX-PLGA-PEG micelles or targeted micelles were measured by using ultraviolet spectrophotometry method, and stimulated release in vitro experiment was done. After administration of 2 mg/ml DOX-PLGA-PEG or targeted micelles, cell morphology change of liver cancer HepG2 and Huh7 was observed by using the phase-contrast photomicrography. After administration of 1 mg/ml DOX-PLGA-PEG or targeted micelles, cell survival was analyzed by using plate clone formation assay.@*Results@#The spectrum peak was around 1 575 cm-1 under the observation of FTIR, which was accord with the location of the peak of amido bond. Activating with p-nitrophenyl chloroformate, DOX was covalently conjugated to PLGA-PEG to produce DOX-PLGA-PEG via a carbamate linkage between the primary amine group in DOX and the terminal hydroxyl group in PLGA-PEG. DLS measurements showed that the diameter of DOX-PLGA-PEG micelles and targeted micelles was (55.0±6.3) nm and (87.6±9.3) nm, respectively, and polydispersity index was 0.098 and 0.142, respectively. SEM micrographs revealed that these nanomicelles had a spherical morphology and relatively smooth surface. Drug loading rate of DOX-PLGA-PEG micelles and targeted micelles was (2.4±0.2)% and (2.2±1.9)%, and the entrapment rate was (91.7±5.3)% and (87.5±4.8)%, respectively. The drug release curve in vitro of DOX-PLGA-PEG micelles and targeted micelles exhibited a biphasic pattern characterized by a fast initial release, followed by a slower and continuous release. The amount of the drug release rate was about 30% within 5 d, and 25% within 6 h. After 2 mg/ml DOX-PLGA-PEG micelles and targeted micelles, the cell morphology of liver cancer HepG2 and Huh7 had the impaired change, and the part of the cells were dead, the clonality decreased. The effect of targeted micelles was more significant compared with DOX-PLGA-PEG micelles. After DOX-PLGA-PEG micelles and targeted micelles, the survival rates of HepG2 and Huh7 cells were decreased with time, and the effect of targeted micelles was more effective compared with DOX-PLGA-PEG micelles (all P < 0.05). The 50% effective inhibition of the targeted micelles and DOX-PLGA-PEG micelles was obtained in 2.4 d and 5.5 d, respectively for Huh7 cells. At these time points, DOX concentration was 1.15 μg/ml and 1.24 μg/ml, respectively. The 50% effective inhibition of targeted micelles and DOX-PLGA-PEG micelles was obtained in 3.3 d and 7.4 d, respectively for HepG2 cells. At these time points, DOX concentration was 1.20 μg/ml and 1.31 μg/ml.@*Conclusion@#DOX nanomicelles with dual effects of passive and active targeting can release a large number of active drugs in vitro, which plays an obvious inhibitory role in the cell proliferation of liver cancer HepG2 and Huh7 cells.
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Objective To investigate the indications and complications of onlay island flap urethroplasty in hypospadias.Methods Retrospective analysis 51 cases who received the onlay urethroplasty from Mar.2005 to Sep.2012 at the First Affiliated Hospital of Guangxi Medical University.Their ages were from 2 years to 12 years,the median age was 4 years and 3 months.Forty-three cases received the operation for the first time,8 cases were re-operation.The prepuce island flaps were implemented in all cases.Factors which contributed to the successful operation ratio were compared,such as the levels of penile curvature,the lengths of urethral defects,first operation or reoperation,the size of glans.Results The operation achievement ratio was 82.4％ (42/51 cases).There were 7 cases urethrocutaneous fistula(6 cases were then cured by reoperation,1 case healed naturally).One case had urinary meatus had retraction.There was another case with persisting penile curvature.Some differences in factors had no statistical significance,such as the levels of penile curvature,the lengths of urethral defects,first operation or reoperation (x2 =0.249,0.287,1.208 ; all P ＞ 0.05).The difference in glans size had statistical significance (x2 =3.393,P ＜ 0.05).Conclusions Onlay urethroplasty was suitable for the cases of coronary sulcus of penis and extremitas anterior penis,but for proximal cases,this therapy must be applied very carefully because of persisting penile curvature.For some reconstruction cases,Onlay urethroplasty was suitable,but the island flap must be designed flexibly.Urethrocutaneous fistula was the most complication,especially in some small flat glans.
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OBJECTIVE:To facilitate the development and popularization of Office Information Network in hospital pharmacy department and provide the carrier and space for developing hospital pharmaceutical services. METHODS: In view of the requirement of hospital pharmacy, B/S structure, ASP technique and MYSQL open source database were used. RESULTS: The system was characterized by concise in style, simple in operation, characteristic and full of affinity, and it has been gradually accepted and widely used by the staffs. CONCLUSIONS: The Office Information Network of hospital pharmacy department can reflect in time the latest development of pharmacy, and it has facilitated the construction and development of informatization in hospital pharmacy department and helped to improve the administrators’ ability in information management and decision-making.
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Objective: To select the prescription of Harmine HCl ointment. Methods: The orthogonal design was used for selecting the prescription with transdermal absorption rate constant (K) and flow energy of activition (E ?) as selecting standard. Results: The optimum prescription is as follows: Azone (2.0%), Span-80 ( 0.2%), Tweens-80 (0.4%), Glycerylmonostearate (2.5%), Vaselin (4.0%), Liquid (11%). Conclusion: The prescription design is available, and the ointment has a good stability.