1. Research on separation and purification of extract from Dracocephalum moldevica with macroporous resins
Chinese Traditional and Herbal Drugs 2016;47(4):599-605
Objective: To investigate the technology for the separation and purification of extract in Dracocephalum moldevica (EDM) by macroporous resin. Methods: Static and dynamic adsorption-desorption were used to select the best one from seven different type macroporous resins; With the content of total flavonoids, tilianin, luteolin-7-O-β-D-glucuronide, and rosmarinic acid as indexes, the purification technology parameters of EDM were optimized. Results: HPD600 resin showed the best purifying profile, its optimum technology conditions were as follows: The optimum concentration of the sample liquid was 0.08 g/mL equivalent to raw material, the resin column diameter-height ratio was 1:9, the amount of used adsorption was 0.32 g dried medicinal herb/mL resin, sample flow rate was 1.5 BV/h, and adsorption time was 12 h. In the course of elution, the resin column chromatography was eluted with 6 BV of 70% ethanol after removing impurities with 4 BV of water by flow rate of 1.5 BV/h. The contents of total flavonoids, tilianin, luteolin-7-O-β-D-glucuronide, and rosmarinic acid were more than 53%, 5.5%, 4.7%, and 2.5%. Conclusion: Macroporous resin HPD600 is suitable to separate and purify EDM.
2. Simultaneous determination of five effective components in different parts of Dracocephalum moldevica L by RP-HPLC
Chinese Pharmaceutical Journal 2016;51(7):583-587
OBJECTIVE: To develop an RP-HPLC method for simultaneous determination of rosmarinic acid, tilianin, luteolin-7-O-β-D-glucuronide, apigenin-7-O-β-D-glucuronide, and diosmetin-7-O-β-D-glucuronide in different parts ofDracocephalum moldevica L. METHODS: The five constituents were measured on a Shim-pack ODS column(4.6 mm×250 mm, 5 μm)with gradient elution of acetonitrile (A)-0.5% formic acid aqeous solution (B) (0-30 min, 17%A;30-60 min, 17%-28%A; 60-70 min, 28%A) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 330 nm, and the column temperature was maitained at 35℃. RESULTS: The linear ranges of rosmarinic acid, tilianin, luteolin-7-O-β-D-glucuronide, apigenin-7-O-β-D-glucuronide, and diosmetin-7-O-β-D-glucuronide were 4.2-126 μg·mL-1 (r=0.999 2), 7.84-235.2 μg·mL-1 (r=0.999 3), 3.048-91.44 μg·mL-1(r=0.999 4), 1.472-44.16 μg·mL-1(r=0.999 4), and 2.816-84.48 μg·mL-1 (r=0.999 2), respectively. The average recoveries (RSD) of the five compounds were 98.97%(1.03%), 99.90%(0.92%), 99.89% (1.75%), 99.55% (0.98%), and 99.76%(1.19%) (n=6), respectively. CONCLUSION: The developed method is accurate and precise, which can be used for the quality control of different parts of Dracocephalum moldevica L.The RESULTS: of content determination indicate that the five compounds exist in all the parts of Dracocephalum moldevica L., but the mass fractions are obviously different.
3. Simultaneous determination the contents of phenylpropanoids and flavonoids in Dracocephalum moldavica L. extract
Chinese Pharmaceutical Journal 2015;50(13):1138-1141
OBJECTIVE: To establish an HPLC method for simultaneous detemination the contents of rosmarinic acid (phenylpropanoids), luteolin-7-O-β-D-glucuronide and tilianin (flavonoids) in Dracocephalum moldavica L. Extract. METHODS: The samples were separated on Purospher® STARLP RP-18 endcapped column(4.6 mm × 250 mm, 5 μm) by gradient elution with acetonitrile (A)-0.5% formic acid solution(B) (0-28 min, 20% A; 28-55 min, 20%→28% A; 55-80 min, 28%→30% A)as the mobile phase at a flow ratio of 1.0 mL · min-1. The detection wavelength was set at 330 nm and the column temperature was maintained at 35℃. RESULTS: The calibration curves of rosmarinic acid, luteolin-7-O-β-D-glucuronide, tilianin were in good linearity over the ranges of 2.184-21.84 μg · mL-1 (r =0.999 6), 3.14-31.84 μg · mL-1(r =0.999 6), 7.9-39.5 μg · mL-1(r =0.999 5) respectively. The average recoveries were 99.62%, 99.64% and 100.12% with RSD values of 1.27%, 1.05% and 1.09%. CONCLUSION: The method is reliable, simple, and accurate, and can be used for the comprehensive quality control of Dracocephalum moldavica L. extract.
4. Extraction technology for active constituents in Dracocephali Moldavici Herba and comparison on contents of Dracocephali Moldavici Herba from various habitats
Chinese Traditional and Herbal Drugs 2015;46(6):846-852
To study the extraction technology for the active constituents in Dracocephali Moldavici Herba (the aerial parts of Dracocephalum moldevica) and to compare their contents Dracocephali Moldavici Herba from various habitats. The contents of luteolin-7-O-glucuronide (I), apigenin-7-O-glucuronide (II), rosmarinic acid, diosmetin-7-O-glucuronide (III), tilianin, and acacetin-7-O-glucuronide (IV) in Dracocephali Moldavici Herba were measured using HPLC method. Based on single-factor test, and the influence of the extracting menstrua, menstruum dosage, and extracting time were investigated using orthogonal design method. Based on the optimal technology, the contents of I, II, rosmarinic acid, III, tilianin, and IV in Dracocephali Moldavici Herba from various habitats were compared. The optimal conditions for the extracting for one time, each time for 5 h with 50 times of amount of 40% ethanol. D. moldevica produced in Jimusar, Xinjiang had the higher contents of I, II, rosmarinic acid, III, tilianin, and acacetin-7-O-glucuronide. This extraction technology is reasonable, stable, and feasible. The contents of I, II, rosmarinic acid, III, tilianin, and IV in Dracocephali Moldavici Herba from different habitats have some differences. The multi-index contents of I, apigenin-7-O-glucuronide, rosmarinic acid, III, tilianin, and IV could reflect the quality of Dracocephali Moldavici Herba more comprehensively.
5. Chemometric Analysis of HPLC Fingerprint of Triterpenoids in Different Ganoderma Preparations Based on Reference Drug
Chinese Pharmaceutical Journal 2020;55(4):298-304
OBJECTIVE: To establish a method combining triterpenoids fingerprint with chemometric analysis based on reference drug, and determine 89 batches of samples to evaluate the quality of Ganoderma preparations. METHODS: The samples were extracted by ultrasonic with methanol, chromatography was performed on Agilent TC C18(2) column (4.6 mm×250 mm, 5 μm) with gradient elution with acetonitrile and 0.02% phosphoric acid aqueous solution. The detection wavelength was set at 254 nm. Principal component analysis and HCA heatmap were used for data analysis. RESULTS: The chromatographic fingerprint has 56 characteristic peaks. By comparing the samples with the reference drug, it was found that Ganoderma preparations were produced using Ganoderma lucidum by almost all manufacturers. Principal component analysis, HCA heatmap analysis and similarity analysis divided the samples into three categories. The first type had characteristic peaks and peak areas which were consistent with the reference drug, and the quality was better. The second type had characteristic peaks which consistent with the reference drug, but the peak areas were smaller and the quality was medium. And the third type was different from the reference drug, which was the problem sample. The strong characteristic peaks for classification of different samples were peak 26, peak 31, peak 24, peak 32 and peak 18. CONCLUSION: The method is comprehensive, accurate and specific, and it can be used for comprehensive quality evaluation of triterpenoids in Ganoderma preparations.
6. Application of hot-melt extrusion technique in pharmaceutical research
Journal of International Pharmaceutical Research 2014;41(4):437-443
Hot-melt extrusion (HME) is the process of pumping raw materials with a rotating screw under the elevated temperature and pressure through a die to from a product with the uniform shape. This technique combines many advantages of solid dispersion technology and mechanical preparation, including dust reduction, short duration of extrusion process, without use of organic solvents and water, without heat drying and hydrolysis, etc. In addition, it has the superiority in solubility, taste masking, drug stability and drug release. Besides, this technique has good reproducibility, high production efficiency, and it can be online monitored in the mass production. Therefore, this technique is widely used in the research and development of drug dosage forms involved in rapid and immediate drug release, sustained drug release, gastrointestinal drug site-specific release, transdermal drug delivery systems and so on. This paper reviews the advantages of HME and its recent application in the pharmaceutical research.
7. The anti-inflammation effects of Aesculetin from Viola tianshanica Maxim via NF-kB and MAPK signaling pathways in RAW 264.7 ceils stimulated by LPS and its mechanism
Xue WANG ; Yan LIU ; Yu-Zhu SHI
Chinese Pharmacological Bulletin 2022;38(9):1340-1349
Aim To investigate the anti-inflammatory effects of Aesculetin from Viola tianshanica Maxim in LPS-stimulated RAW 264.7 cells and the underlying mechanism.Methods RAW 264.7 cells were divided into control group, model group( LPS), 0.16, 0.8, 4, 20 μmol·L-1 AESN groups( different concentrations of AESN + LPS)and positive control group(10 μmol·L-1 Indomethacin+LPS).LPS(1 mg·L-1)was used to stimulate RAW 264.7 cells for 24 h to establish inflammatory model.MTS assay was used to detemine cytotoxicity of Aesculetin in RAW 264.7 cells.Griess method was used to detect NO secretion in LPS-stimulated RAW 264.7 cells.ELISA was applied to determine the contents of TNF-α, IL-6 and IL-1β in cell culture supernatant.qRT-PCR was employed to detect the mRNA expression levels of TNF-α, IL-6, IL-1β and iNOS.Immunofluorescence assay was used to evaluated the protein expressions of iNOS, p-NF-κB p65, IκBα, p-p38 and p-ERK1/2.Enzyme assay was used to detect the inhibition activity of Aesculetin on cyclooxygenase 1/2(COX 1/2).Results Aesculetin significantly inhibited the secretion of inflammatory mediator NO, mRNA and protein expression of iNOS in LPS-induced RAW 264.7 at 0.16, 0.8, 4 and 20 μmol·L-1.The contents of TNF-α, IL-6 and IL-1β in supernatant significantly decreased, and the mRNA expression levels of TNF-α, IL-6 and IL-1β were also reduced by Aesculetin.Aesculetin also obviously inhibited the protein degradation of IκBα and inhibited the nuclear translocation of p-NF-κB p65, p-p38, p-ERK1/2.In addition, Aesculetin had significant inhibitory activities on COX-1 and COX-2, and the IC50 was 28.1 μmol·L-1, 2.3 μmol·L-1, respectively.Conclusions AESN has good anti-inflammatory effect, and its mechanism is closely related to the inhibition of NF-κB and MAPK signaling pathways
8.Analysis on monitoring results of heavy metals in rural soils in Bole of Xinjiang, 2017-2019
Journal of Public Health and Preventive Medicine 2020;31(2):19-22
Objective To monitor heavy metals Pb, Cd, Cr, As, and Hg in agricultural land in Bole of Xinjiang, in order to understand the overall pollution status of rural soil environment and determine the environmental health hazards in rural areas of Bozhou, Xinjiang. Methods From 2017 to 2019, 4 administrative villages in 5 townships in Bole City in Bozhou Region were selected as monitoring points. The 5 to 20 cm deep topsoil of monitoring points was collected for a total of 60 samples. According to "Environmental Quality Standard for Soils" (GB 15618-2018), the pollution levels of Pb, Cd, Cr, As and Hg were evaluated by single index method and Nemerow index method. Results The content of Pb, Cr, As, and Hg in the heavy metals in the soil of Bozhou region from 2017 to 2019 were all lower than the screening value and regulatory value of soil pollution risk for agricultural land, while the content of heavy metal Cd in 13 soil samples (21.67%) exceeded the standard. The content difference in Pb, Cd, Cr, and As was statistically significant (P<0.05), while the difference in Hg was not statistically significant (P>0.05). The single pollution index Pi of Pb, Cr, As and Hg was less than 1, indicating a non-pollution state. However, the single pollution index Pi of Cd from 2017 to 2018 was higher than 1, with the highest value reaching 2.33. In 2017 and 2018, the comprehensive pollution index of rural soil samples was 1.67 and 1.06, and the evaluation result was light pollution, while in 2019, the comprehensive pollution index was 0.72, and the evaluation result was still clean. Conclusion The overall quality of rural environmental soil in some rural areas in Bozhou, Xinjiang was generally acceptable, but the heavy metal Cd pollution in some aeras deserves attention.
9.Progress of Keystroke Dynamics in Deception Research.
Journal of Forensic Medicine 2022;38(6):783-787
Traditional polygraph techniques mostly rely on the changes of an individual's physiological indicators, such as electrodermal activity, heart rate, breath, eye movement and function of neural signals and other indicators. They are easily affected by individual physical conditions, counter-tests, external environment and other aspects, and it is difficult to conduct large-scale screening tests based on the traditional polygraph techniques. The application of keystroke dynamics to polygraph can overcome the shortcomings of the traditional polygraph techniques to a large extend, increase the reliability of polygraph results and promote the validity of legal evidence of polygraph results in forensic practice. This paper introduces keystroke dynamics and its application in deception research. Compared with the traditional polygraph techniques, keystroke dynamics can be used with a relatively wider application range, not only for deception research but also for identity identification, network screening and other large-scale tests. At the same time, the development direction of keystroke dynamics in the field of polygraph is prospected.
Lie Detection
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Reproducibility of Results
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Forensic Medicine
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Deception
10.Effect of LAG3 deficiency on natural killer cell function and hepatic fibrosis in mice infected with Echinococcus multilocularis
Rousu ZIBIGU ; Ainiwaer ABIDAN ; Duolikun ADILAI ; Yinshi LI ; Xuejiao KANG ; Qian YU ; Bingqing DENG ; Xuran ZHENG ; Maolin WANG ; Jing LI ; Hui WANG ; Chuanshan ZHANG
Chinese Journal of Schistosomiasis Control 2024;36(1):59-66
Objective To investigate the effect of LAG-3 deficiency (LAG3-/-) on natural killer (NK) cell function and hepatic fibrosis in mice infected with Echinococcus multilocularis. Methods C57BL/6 mice, each weighing (20 ± 2) g, were divided into the LAG3-/- and wild type (WT) groups, and each mouse in both groups was inoculated with 3 000 E. multilocularis protoscoleces via the hepatic portal vein. Mouse liver and spleen specimens were collected 12 weeks post-infection, sectioned and stained with sirius red, and the hepatic lesions and fibrosis were observed. Mouse hepatic and splenic lymphocytes were isolated, and flow cytometry was performed to detect the proportions of hepatic and splenic NK cells, the expression of CD44, CD25 and CD69 molecules on NK cell surface, and the secretion of interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), interleukin (IL)-4, IL-10 and IL-17A. Results Sirius red staining showed widening of inflammatory cell bands and hyperplasia of fibrotic connective tissues around mouse hepatic lesions, as well as increased deposition of collagen fibers in the LAG3-/-group relative to the WT group. Flow cytometry revealed lower proportions of mouse hepatic (6.29% ± 1.06% vs. 11.91% ± 1.85%, P < 0.000 1) and splenic NK cells (4.44% ± 1.22% vs. 5.85% ± 1.10%, P > 0.05) in the LAG3-/- group than in the WT group, and the mean fluorescence intensity of CD44 was higher on the surface of mouse hepatic NK cells in the LAG3-/- group than in the WT group (t = −3.234, P < 0.01), while no significant differences were found in the mean fluorescence intensity of CD25 or CD69 on the surface of mouse hepaticNK cells between the LAG3-/- and WT groups (both P values > 0.05). There were significant differences between the LAG3-/- and WT groups in terms of the percentages of IFN-γ (t = −0.723, P > 0.05), TNF-α (t = −0.659, P > 0.05), IL-4 (t = −0.263, P > 0.05), IL-10 (t = −0.455, P > 0.05) or IL-17A secreted by mouse hepatic NK cells (t = 0.091, P > 0.05), and the percentage of IFN-γ secreted by mouse splenic NK cells was higher in the LAG3-/- group than in the WT group (58.40% ± 1.64% vs. 50.40% ± 4.13%; t = −4.042, P < 0.01); however, there were no significant differences between the two groups in terms of the proportions of TNF-α (t = −1.902, P > 0.05), IL-4 (t = −1.333, P > 0.05), IL-10 (t = −1.356, P > 0.05) or IL-17A secreted by mouse splenic NK cells (t = 0.529, P > 0.05). Conclusions During the course of E. multilocularis infections, LAG3-/- promotes high-level secretion of IFN-γ by splenic NK cells, which may participate in the reversal the immune function of NK cells, resulting in aggravation of hepatic fibrosis.