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Aim To establish the 3D hepatocyte model by selecting the humanized hepatocyte HepG2 cells and 3D cell culture methods, and to establish the 3D hepatocyte cytokinesis-block micronucleus cytome(CBMN-cyt)assay and 3D hepatocyte comet assay by using chemicals of different mode of action.Methods In this study, a scaffold-free culture method was used to successfully establish a 3D HepG2 hepatocyte spheroid model.The appearance of the sphere, the survival rate of cells inside the sphere, the gene expression of phase I and II metabolic enzymes, and the expression of liver-specific biomarkers were selected as the observation indicators to obtain the best culture conditions for the 3D hepatocyte model.The 3D hepatocyte model was combined with in vitro micronucleomics test and in vitro comet test to explore its applicability for genotoxicity test.Results The best culture conditions for the 3D hepatocyte model was 5×103 cells/20 μL /drop inoculation, cultivating for seven days.A 3D hepatocyte CBMN-cyt assay was established using mitomycin C(MMC), a micronucleus positive compound, and the results showed that it could successfully detect the genotoxicity and cytotoxicity of MMC.Compared with the CBMN-cyt results of 2D hepatocyte model, 3D hepatocyte model had higher sensitivity in detecting MN and Nbud.The 3D hepatocyte comet assay methods were established using the known in vivo and in vitro comet assay positive compound methyl methanesulfonate(MMS), and the results showed that MMS could significantly increase the tail DNA% of 3D hepatocytes with low cytotoxicity.The sensitivity of 3D hepatocyte model to MMS genotoxicity detection was higher than that of 2D cells.Conclusions The 3D hepatocyte model established in this study is easy to use and low in cost, and shows good sensitivity and specificity in the in vitro micronucleus test and comet test, suggesting that the 3D hepatocyte genotoxicity test method is used in early drug genotoxicity screening.It has good application prospects in additional experimental research.
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Objective::Ultra performance liquid chromatography (UPLC) fingerprint was used to evaluate the storage effect of different packaging and storage conditions on Notopterygh Rhizoma et Radix pieces. Method::UPLC fingerprints of Notopterygh Rhizoma et Radix pieces with different storage time and packaging conditions were established under following chromatographic conditions: Waters UPLC BEH C18 column (2.1 mm×50 mm, 1.7 μm), mobile phase 0.3%phosphoric acid solution(A)-acetonitrile (B) for gradient elution (0-4 min, 90%-47%A; 4-8 min, 47%A, ; 8-12 min, 47%-20%A; 12-16 min, 20%A; 16-18 min, 20%-90%A), flow rate of 0.2 mL·min-1, column temperature at 35 ℃, detection wavelength of 246 nm, and injection volume of 1.0 μL. The fingerprints were evaluated in terms of similarity, cluster analysis and principal component analysis. Result::The similarity evaluation showed that the UPLC fingerprint patterns of Notopterygh Rhizoma et Radix pieces with different storage time and different packaging and storage treatments were basically the same, and the similarity was above 0.95.Systematic clustering shows that Notopterygh Rhizoma et Radix pieces packaged in plastic bags, stored under light and shade and the initial live-action pieces were clustered into one group. Principal component analysis showed that Notopterygh Rhizoma et Radix pieces packaged in plastic bags, stored under light and shade had the highest comprehensive scores. Conclusion::Storage time, packaging material and storage temperature will not cause the increase or decrease of internal components of Notopterygh Rhizoma et Radix, but only affect the content of specific components. Plastic bag packaging, light and cool storage conditions are more suitable for the preservation of non-volatile oil components of Notopterygh Rhizoma et Radix.
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Objective::To study the effect of different packaging methods and storage conditions on the quality of Notopterygii Rhizoma et Radix pieces, in order to determine the optimal packaging method and suitable storage conditions for Notopterygii Rhizoma et Radix pieces. Method::Different packaged Notopterygii Rhizoma et Radix pieces were stored in different environments in a one-year long-term stability experiment. The appearance, water content, extract content and volatile oil content of Notopterygii Rhizoma et Radix pieces were regularly observed. Result::During the 1-year storage period, the Notopterygii Rhizoma et Radix pieces under different packaging and storage conditions showed different degrees of quality changes. Among them, the samples packed in polyethylene plastic bags and polyethylene aluminum foil composite bags were better preserved. The fluctuations in water content of the sample packed in polyethylene plastic bags were relatively low, and the RSD value of water content during the month was less than 11.5%. The extracts and volatile oil contents of each sample were reduced to different degree, but the samples packed in plastic sealed bags and protected from light had the smallest annual loss of extracts (1.27%), with the lowest monthly loss rate of volatile oil (0.08%). Conclusion::The quality of Notopterygii Rhizoma et Radix pieces can be well preserved in plastic sealed bags and storage in dark and cool conditions, which is suitable for the storage of Notopterygii Rhizoma et Radix pieces.
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Due to the large amount of nutrients required during the cultivation of Angelica sinensis and in order to prevent the occurrence of pests and diseases,and the annual reduction of the planting area of Angelica and the balance of supply and demand of A. sinensis,the A. sinensis plantation adopts the rotation mode. This paper takes Wuyuan county of Gansu province as the research scope and use GF-1 Satellite data as the data source,using remote sensing technology combined with field survey results,to explore the effective method of visual interpretation for the extraction of A. sinensis planting area. A sample was selected to generate a spectrum according to different feature types. The different characteristics of A. sinensis and other features were analyzed and distinguished in remote sensing images,so that the A. sinensis planting plots were extracted and verified in remote sensing images. The results showed that the accuracy verification value of the visual interpretation method was 95. 85%. It is determined that the visual interpretation method can effectively extract the A. sinensis planting plots within the research scope and realize the comprehensive grasp of the spatial distribution information of A. sinensis.
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Angelica sinensis , China , Plants, Medicinal , Remote Sensing TechnologyABSTRACT
Due to the large amount of Codonopsis pilosula planted in Weiyuan county,and the arable land area,the local medicinal materials office uses a large amount of manpower,financial resources and material resources to estimate its area every year. In order to extract the information of local Chinese medicinal materials more quickly and simply,we try to apply remote sensing technology to the extraction of Chinese medicinal materials. This paper will use Weiyuan county of Gansu province as the research area,and use the domestic ZY-3 Satellite multi-spectral remote sensing image as the data source to find out the spectral characteristics of the party's participation in other remote sensing images. The visual interpretation method was used to extract the planting area of the C. pilosula in Weiyuan county. The estimated value of the planting area of C. pilosula using satellite remote sensing technology was 75 965 mu( 1 mu≈667 m2),which was basically consistent with the field survey data of the local medicinal materials office. After the accuracy verification,it was found that the precision of C. pilosula planted by visual interpretation was more than 70%. It is concluded that the satellite remote sensing technology can be used to extract the information of C. pilosula and it can provide the relevant information of the planting area of Chinese medicinal materials quickly and accurately.
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China , Codonopsis , Plants, Medicinal , Remote Sensing TechnologyABSTRACT
Objective To investigate the therapeutic effects of penetrating resin and fluoride on early enamel caries. Methods Sixty intact bovine incisors were immersed in demineralized solution for 24 hours to make bovine incisor enamel caries.The specimens were divided into four groups(n=15 for each group)according to the treatment methods:control group (CON)-immersion in artificial saliva, DF group-immersion in 0.05% fluoride solution daily, WF group-2% fluoride gel weekly and IC group-resin infiltration.After processing for four and eighe weeks,the microhardness of the surface of each group was measured.After the treatment for four weeks,the depth of penetration and the microhardness of the samples were measured. After 8-week treatment, all samples were reintroduced into the demineralized solution for 24 hours and the microhardness of the samples was measured again.Results Results of microhardness assessment showed that there were no significant differences in baseline values(after white spots)between four groups(P>0.05).After treatment for four weeks the microhardness value reached the peak in IC group. After treatment for eight weeks the microhardness values reached the peak in DF group and WF group.The values of microhardness were significantly higher at different time points in IC group than those of other groups(P<0.05).After 4-week treatment,the percentages of penetration depth were significantly higher in DF,WF and IC groups than those of control group(P<0.01).The penetration depth was significantly higher in IC group than that of DF group and WF group (P<0.05). There was no significant difference in the penetration depth between DF group and WF group(P>0.05).Conclusion For the early enamel caries penetration resin,the penetration percentage is significantly higher than the fluoride treatment.