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1.
Article in Chinese | WPRIM | ID: wpr-1017316

ABSTRACT

Objective:To discuss the inhibitory effect of chelerythrine(CHE)on the migration,invasion,and epithelial-mesenchymal transition(EMT)of the human ovarian cancer SKOV3 cells,and to clarify the associated mechanism.Methods:The SKOV3 cells were cultured in vitro and divided into control group and 2.5,5.0,10.0,20.0,and 40.0 μmol·L-1 CHE groups.Methylthiazolydiphenyl-tetrazolium(MTT)assay was used to detect the inhibitory rates of proliferation of the cells in various groups.The SKOV3 cells were cultured in vitro and divided into control group,transforming growth factor-β1(TGF-β1)group,TGF-β1+5 μmol·L-1 CHE group,and TGF-β1+10 μmol·L-1 CHE group.Cell scratch assay was used to detect the migration rates of the cells in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion cells in various groups;Western blotting method was used to detect the expression levels of E-cadherin,N-cadherin,and Vimentin proteins in the cells in various groups;immunofluorescence staining method was used to detect the fluorescence intensities of E-cadherin and N-cadherin in the cells in various groups.Results:The MTT assay results showed that compared with control group,the inhibitory rates of proliferation of the cells in 5.0,10.0,20.0,and 40.0 μmol·L-1 CHE groups were significantly increased(P<0.05 or P<0.01).The cell scratch assay results showed that compared with control group,the migration rate of the cells in TGF-β1 group was increased(P<0.01);compared with TGF-β1 group,the migration rates of the cells in TGF-β1+5 μmol·L-1 CHE group and TGF-β1+10 μmol·L-1 CHE group were significantly decreased(P<0.01).The Transwell chamber assay results showed that compared with control group,the numbers of migration and invasion cells in TGF-β1 group were significantly increased(P<0.05);compared with TGF-β1 group,the numbers of migration and invasion cells in TGF-β1+5 μmo·l L-1 CHE group and TGF-β1+10 μmo·l L-1 CHE group were significantly decreased(P<0.01).The Western blotting results showed that compared with control group,the expression level of E-cadherin protein in the cells in TGF-β1 group was significantly decreased(P<0.01),while the expression levels of N-cadherin and Vimentin proteins were increased(P<0.05 or P<0.01);compared with TGF-β1 group,the expression levels of E-cadherin protein in the cells in TGF-β1+5 μmol·L-1 CHE group and TGF-β1+10 μmol·L-1 CHE group were significantly increased(P<0.01),and the expression levels of N-cadherin and Vimentin proteins were significantly decreased(P<0.01).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of E-cadherin in the cells in TGF-β1 group was decreased,and the fluorescence intensity of N-cadherin was increased;compared with TGF-β1 group,the fluorescence intensities of E-cadherin in the cells in TGF-β 1+5 μmol·L-1 CHE group and TGF-β1+10 μmol·L-1 CHE group were significantly increased,and the fluorescence intensities of N-cadherin were decreased.Conclusion:CHE can inhibit the proliferation,migration,invasion,and EMT of the human ovarian cancer SKOV3 cells.

2.
China Pharmacy ; (12): 1189-1197, 2022.
Article in Chinese | WPRIM | ID: wpr-924071

ABSTRACT

OBJECTIVE To prepare hyperoside mixed nanomicelles (Hyp-F127/TPGS) and optimize its preparation technology,and to investigate its intestinal absorption characteristics. METHODS Hyp-F127/TPGS was prepared by thin film dispersion method. Based on single factor test and Plackett-Burman design ,combined with Box-Behnken response surface method , the preparation process was optimized and validated using entrapped efficiency (EE)and drug loading (DL)as evaluation indexes , F127-TPGS mass ratio ,hydration time and the amount of Hyp as factors. The appearance and microscopic morphology of Hyp-F127/TPGS obtained by the optimal technology were observed ,and the particle size ,polydispersity index (PDI)and Zeta potential were also determined. The critical micelle concentration (CMC)of blank micelle (F127/TPGS),in vitro release behavior and preliminary stability of Hyp-F 127/TPGS were investigated ,and absorption characteristics of Hyp-F 127/TPGS were investigated by in situ unidirectional intestinal perfusion model. RESULTS The optimal preparation technology of Hyp-F 127/TPGS included F127-TPGS mass ratio of 2∶1,hydration time of 2 h,and Hyp amount of 9 mg. Results of three validation tests showed that the EE of Hyp-F 127/TPGS was (87.20±0.99)%,and the DL was (5.02±1.20)%,deviations from predicted values were 0.92% and 2.39%. The micelles prepared by optimal technology were yellow ,clear and transparent solution ,with good Tyndall effect ;under transmission electron microscope ,they were spherical ,complete and evenly distributed ;the particle size was (15.02±0.16)nm, the PDI was 0.092±0.031,and the Zeta potential was (-6.67±1.47)mV. The CMC of F 127/TPGS was 21 μg/mL,Hyp-F127/ TPGS was stable after 4 weeks of storage at 4 ℃,and the cumulative release rates of Hyp-F 127/TPGS and Hyp control were (66.30±2.93)%(96 h)and(99.24±0.27)%(60 h),respectively. Hyp-F 127/TPGS and Hyp reference were absorbed in each intestinal segment ,and the main absorption sites were jejunum and duodenum respectively ;drug absorption rate constant andapparent absorption coefficient of the former were significantly higher than those of the latter (P<0.05 or P<0.01). E-mail:zhangyuhangxz@163.com CONCLUSIONS The optimized preparation technology of Hyp-F127/TPGS is stable and feasible ;prepared Hyp-F 127/ TPGS shows a sustained -release effect ,which promotes the intestinal absorption of H yp to a certain extent.

3.
China Pharmacy ; (12): 1103-1107, 2020.
Article in Chinese | WPRIM | ID: wpr-821501

ABSTRACT

OBJECTIVE:To investigate t he effects of different compatibility ratio of Gardenia jasminoides to fermented soybean on the content of genistein and total flavonoids ,and to investigate the compatibility regularity of Zhizichi decoction. METHODS:The decoction method was used to prepare the mixed decoction with different compatibility ratio of G. jasminoides to fermented soybean (2∶1,1∶1,1∶2,1∶4,m/m,the same hereinafter ). UPLC-MS/MS method was used to determine the content of genistein in Zhizichi decoction with different compatibility ratio and corresponding fermented soybean single decoction. UV method was used to determine the content of total flavonoids in Zhizichi decoction with different compatibility ratio and corresponding gardenia single decoction and fermented soybean single decoction. RESULTS :The established method had good linearity , precision,repeatability,stability and accuracy. Compared with single decoction ,the content of genistein in the mixed decoction with different compatibility ratio of G. jasminoides to fermented soybean (2∶1,1∶1,1∶2,1∶4)was decreased to different extents , while the content of total flavonoids was increased to different extents. With the increase of fermented soybean ,the content of genistein in the decoction increased at first and then decreased. When the compatibility ratios of G. jasminoides to fermented soybean were 1 ∶ 1 and 1 ∶ 2,the content of genistein in the decoction was the highest (all 0.071 μg/mL). With the increase of fermented soybean ,the content of total flavonoids in the decoction did not change regularly ;when the ratio of G. jasminoides to fermented soybean was 1 ∶ 1,the content of total flavonoids in the decoction was the highest (1.861 μg/mL). CONCLUSIONS : When the compatibility ratio of G. jasminoides to fermented soybean was 1 ∶ 1,the content of flavonoids in the decoction is the highest.

4.
China Pharmacy ; (12): 202-206, 2019.
Article in Chinese | WPRIM | ID: wpr-816721

ABSTRACT

OBJECTIVE: To study the chemical constituents of ethyl acetate fraction of Panax ginseng fungal substance obtained by biotransformation, in order to obtain compounds with better activity and lower toxicity, and to provide reference for new drug R&D and the second development and utilization of P. ginseng. METHODS: Fungus of Code Name C-1 seed solution was added into the culture medium containing P. ginseng, and P. ginseng fungal substance was obtained by biotransformation; the dried P. ginseng fungal substance were weighed, extracting with 70% ethanol solvent and concentrating to obtain thick paste. The thick paste was added with water suspension and extracted with ethyl acetate to obtain ethyl acetate fraction. TLC, silica gel column chromatography, ODS column chromatography and semi-prepared liquid phase were used to isolate and purify above ethyl acetate fraction, and the compound structure was identified according to physicochemical properties, hydrogen spectrum (1H-NMR) and carbon spectrum (13C-NMR) data. RESULTS: Eight compounds were isolated and identified from the ethyl acetate fraction of P. ginseng fungal substance and identified as ginsenoside Rs7 (1), ginsenoside Rk3 (2), oleanolic acid-28-O-β-D-glucopyranoside (3), ginsenoside Rs6 (4), 20(R)-ginsenoside Rh1 (5), ginsenoside F1 (6), notoginsenoside R2 (7) and ginsenoside F4 (8). CONCLUSIONS: All the above compounds were found in P. ginseng fungal substance, which compounds 3, 5, 6, 7 and 8 were obtained after biotransformation, proving that biotransformation technology can change the chemical composition of ginseng.

5.
China Pharmacy ; (12): 2546-2551, 2019.
Article in Chinese | WPRIM | ID: wpr-817275

ABSTRACT

OBJECTIVE: To establish the method for simultaneous determination of saikosaponin a and saikosaponin d in Bupleurum chinense water extract, and to optimize its water extraction technology for electromagnetic cracking. METHODS: HPLC method was used. The determination was performed on SB-C18 column with mobile phase consisted of acetonitrile-water (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was 40 ℃. The detection wavelength was set at 210 nm, and  the sample size was 10 μL. Based on single factor experiment, using extraction time, particle size, solide-liquid ratio as factors, total extraction rate of saikosaponin a to saikosaponin d as indexes, the extraction technology was optimized by using Box-Behnken response surface methdology, and compared with the results of ultrasound method and decoction method. RESULTS: The linear range of saikosaponin a and saikosaponin d were 50.70-202.80 μg/mL (r=0.999 9) and 50.50-202.00 μg/mL (r=0.999 9), respectively. The quantitation limits were 0.16 and 0.13 μg/mL, respectively. The detection limits were 0.05 and 0.04 μg/mL,respectively. RSDs of precision, stability and reproducibility tests were all lower than 2%. The average recoveries were 98.23-102.47% (RSD=1.80%, n=6) and 98.84%-102.06% (RSD=1.60%, n=6). The optimal extraction technology was as follows: the extraction time of 2.50 min; the particle size of 80 mesh, solid-liquid ratio of 1 ∶ 28 (g/mL). Results of 3 times of validation tests showed that the optimal technology included the average total extraction rates of saikosaponin a and saikosaponin d were 8.42 mg/g, which was higher than that of ultrasonic method (8.34 mg/g) and decoction method (8.06 mg/g), and the extration time was shorter. CONCLUSIONS: Established method is simple and accurate, and can be used for simultaneous determination of saikosaponin a and saikosaponin d in B. chinense water extract. The optimized water extraction technology for electromagnetic cracking is stable and feasible.

6.
China Pharmacy ; (12): 3413-3418, 2019.
Article in Chinese | WPRIM | ID: wpr-817405

ABSTRACT

OBJECTIVE: To optimize the integrated processing technology for “precise decoction pieces” of Helianthus annuus. METHODS: The contents of total flavonoids and total protein in H. annuus were determined by UV spectrophotometry with rutin and bovine serum albumin as control. Refering to Chinese Pharmacopoeia, the contents of water soluble extract, dilute ethanol extracts and ethyl acetate extract were determined. Based on the different needs such as maintaining quality consistency with the original medicinal materials, preference for anti-gout treatment, preference for liver calming and pain relief, using the contents of total flavonoids, total protein and 3 kinds of polar extract as indexes, gray correlation method was used to optimize the integrated processing technology of 3 kinds of “precise decoction pieces” of H. annuus. RESULTS: Gray correlation analysis showed that the ideal sample sequence of decoction pieces in massive shape dried at 60 ℃ with the original medicinal materials and decoction pieces with preference use of liver calming and pain relief was the most relevant; the ideal sample sequence of ideal sample sequence of decoction pieces in massive shape dried in the shade with decoction pieces with clinical application preference of anti-gout therapy was the most relevant. CONCLUSIONS: Different integrated processing technology for “precise decoction pieces” of H. annuus can be adopted for different needs. If it is necessary to keep the quality consistent with the original medicinal materials or to prepare H. annuus decoction pieces for liver calming and pain relieving, medicinal material can be cut into massive shape and dried at 60 ℃; if it is necessary to prepare H. annuus decoction pieces for anti-gout treatment, cutting into massive shape and drying in the shade can be adopted.

7.
Article in Chinese | WPRIM | ID: wpr-491210

ABSTRACT

Objective To investigate the effect of 1-methylhydantoin (1-MH)on asthma and cough animal model, and to clarify its mechanism preliminarily.Methods 50 Wistar rats with ovalbumin-induced asthma were randomly divided into model group, 1-MH 20, 40, and 80 mg · kg-1 groups, positive control group (aminophylline, 60 mg·kg-1 ),another 10 Wistar rats served as control group.The levels of interleukin-5 (IL-5 ),eosinophil chemotactic factor (Eotaxin)and eosinophil (EOS)count in bronchoalveolar lavage fluid (BALF)were measured in various groups. 40 guinea pigs of asthma were randomly divided into model group, 1-MH 15, 30, and 60 mg·kg-1 groups, positive control group (aminophylline, 50 mg · kg-1 );the asthma incubation period of guinea pig was measured in various groups. The bronchus of 10 guinea pigs were selected and put in Krebs solution,and the antispasmodic percentages against histamine phosphate of 1-MH 0.25,0.50,and 1.00 g ·L-1 were recorded and calculated. 50 mice of cough were randomly divided into model group,1-MH 25,50,and 100 mg·kg-1 groups, positive control group (codeine, 50 mg · kg-1 );the cough incubation period and the number of cough in the mice were measured in various groups.40 guinea pigs of cough were randomly divided into model group,1-MH 15,30,and 60 mg·kg-1 groups,positive control group (codeine,20 mg·kg-1);the cough incubation period and the number of cough of the guinea pigs in various groups were measured. Results Compared with sensitized model control group,the levels of IL-5,Eotaxin and EOS count in BALF of the rats in 1-MH 40 mg·kg-1 and 80 mg·kg-1 groups were reduced (P<0.05 or P<0.01 );compared with acetylcholine-induced asthma model control group, the asthma incubation period of guinea pig in 1-MH 30 mg·kg-1 and 60 mg·kg-1 groups was prolonged(P<0.05 or P<0.01);compared with blank control group, the antispasmodic percentages against histamine phosphate in 1-MH 0.50 g · L-1 and 1.00 g · L-1 groups were increased(P<0.01);compared with mouse cough model control group,the cough incubation period of the mice was prolonged,the cough number of the mice was decreased in 50 mg·kg-1 and 100 mg·kg-1 groups(P<0.05 or P<0.01);compared with guinea pig cough model control group,the cough incubation period of the guinea pig was prolonged,the cough number of the guinea pig was decreased in 1-MH 30 mg·kg-1 and 60 mg·kg-1 groups (P<0.05 or P<0.01).Conclusion 1-MH has good antiasthmatic and antitussive effects,which may be related to inhibition of airway inflammation and relaxation of bronchial smooth muscle directly .

8.
International Journal of Surgery ; (12): 238-242,封3, 2013.
Article in Chinese | WPRIM | ID: wpr-598413

ABSTRACT

Objective To research the effect of mir-520c-3p targeted GPC3 to the hepatocellular carcinoma Huh-7 cell proliferation,migration,and the influence of the attack ability and find new theoretical basis for liver hepatocellular carcinoma clinical treatment.Methods The cells were divided into three groups:not transfection of mir-520c-3p group (cell group),negative control group (Nc group),and transfection of mir-520c-3p group (treat ment group).Then used fluorescence quantitative PCR and Western Blot to detect GPC3mRNA gene and protein expression quantity.Cell proliferation of change was detected by the EDU.Made use of Transwell to detect cell invasion and migration ability of the change.Results Fluorescence quantitative PCR results showed that Cell group,NC group and treatment group were 1.13 ± 0.23,1.28 ± 0.15 and 1.05 ± 0.19 (P > 0.05),mir-520-3p could not reduce the GPC3mRNA; but Western Blot detection results showed that GPC3 protein expression level reduce significantly after transfection mir-520c-3p,Cell,NC and treatment group were 2.16 ± 0.08,1.99 ± 0.04 and 0.499 ± 0.05 (P < 0.01).The EDU detection results showed that hepatocellular carcinoma Huh-7 cell proliferation ability obviously inhibited after transfection mir-520c-3p,Cell group,NC group and treatment group were (90.12 ± 1.93) %,(91.02 ± 0.35) % and (77.73 ± 5.88) % (P < 0.05),and Transwell test found that hepatocellular carcinoma Huh-7cell invasion abilities were restrained,Cell group,NC group and treatment group were 0.071 ±0.008,0.105 ±0.001 and 0.048 ± 0.002 (P < 0.05),in the same the cells' migration abilities were reduced,Cell group,NC group and treatment group were 0.546 ± 0.010,0.328 ± 0.002 and 0.151 ± 0.002 (P <0.01).Conclusions Mir-520c-3p can target GPC3 so that affect hepatocellular carcinoma Huh-7 cell proliferation,invasion and migration abilities.

9.
Article in Chinese | WPRIM | ID: wpr-394444

ABSTRACT

Objective To investigate the feasbility of the orient differentiation from embryo hepatic Sca-1+ cell to hepatic cell or hepatic cell precursor under the induction of hepatocyte growth factor(HGF) and acor fibroblast growth factor(aFGF) in vitro. Methods Sea-1+ cells were isolated by immunomagnetic beads and their morphous were observed by contrast phase microscope. The expression of mRNA of albumin(ALB) and meta-thyroprotein were detected by RT-PCR; ALB, AFP, CK8/18 protein were detected by immunohistochemical method, and cell staining for glycogen and urea synthesis function were tested. Results The activity, purity, recovery rate of Sca-1+ cells were (94. 24±1.04) %, (85.57±1.66) %, (62. 31±1.85 ) % respectively. After the induction of HGF and aFGF, Sca1+ cells became anomalism and transformed to heptic cell in morphous,disappeared in expression of AFP protein and upregulated in expression of ALB and CK8/18 protein,upregulated in expression of ALB and transthyroprotein mRNA,cell staining of glycogen and urea synthesis function were strengthened,the cell differentiated to mature hepatic cell. Conclusion Embryo hepatic Sca-1+ cell could differentiate to hepatic cell or hepatic cell precursor under the induction of HGF and aFGF in vitro.

10.
Journal of Integrative Medicine ; (12): 1286-9, 2008.
Article in Chinese | WPRIM | ID: wpr-450139

ABSTRACT

To extract praeruptorin A from Radix Peucedani by supercritical fluid extraction (SFE)-CO2.

11.
Article in Chinese | WPRIM | ID: wpr-531107

ABSTRACT

0.05).CONCLUSION: The high expression of GPC3 mRNA in hepatocellular carcinoma is independent to the gene mutation,and to the expression of P53 and PCNA protein.

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