1.Exploration of Spectrum-effect Relationship of Zhuriheng Dropping Pills Against Macrophage Foaming Based on UPLC-Q-Exactive Orbitrap MS
Qiong ZHAI ; Fangyuan LIANG ; Yuewu WANG ; Ren BU ; Xin DONG ; Jingkun LU ; Peifeng XUE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):176-186
ObjectiveThrough the correlation analysis between intestinal absorption profile and inhibition of macrophage foaming, the pharmacodynamic components of Zhuriheng dripping pills(ZRH) were explored to provide a basis for establishing its quality standard. MethodIntestinal absorption fluids with 0, 5, 10, 15, 20 times clinical equivalent doses were prepared by a rat everted gut sac(EGS), and the oxidized low density lipoprotein(ox-LDL)-induced RAW264.7 macrophage foaming model was used to investigate the effect of intestinal absorption fluid with different doses on the accumulation of lipids in RAW264.7 cells by oil red O staining and cholesterol content determination, and to screen for the optimal dose. Ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used to analyze and identify intestinal absorption fractions of ZRH intestinal absorption fluids, and partial least squares-discriminant analysis(PLS-DA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were performed on different doses of ZRH intestinal absorption fluids using SIMCA 13.0 with peak area as the independent variable and the pharmacodynamic indicators as the dependent variables to screen the compounds with variable importance in the projection(VIP) value>1.0 as contributing components, and Pearson correlation analysis was used to determine the spectral effect relationship, determined the compounds and positive correlation with pharmacodynamic were as active ingredients. Molecular docking was used to verify the binding energy of peroxisome proliferator-activated receptor α(PPARα), PPARγ, PPARβ, human retinoid X receptor α(RXRA) and nuclear transcription factor-κB(NF-κB) with the active ingredients in ZRH intestinal absorption fluids. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was performed to detect the mRNA levels of PPARγ, scavenger receptor A1(SRA1) and adenosine triphosphate-binding cassette transporter A1(ABCA1) in RAW264.7 cells, Westen blot was used to detect the expression level of PPARγ protein in RAW264.7 cells, and enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of interleukin(IL)-1β and NF-κB in RAW264.7 cells. ResultAccording to the results of oil red O staining and cholesterol content determination, the ZRH intestinal absorption fluids could significantly reduce macrophage foaming, and intestinal absorption fluids with 15, 20 times clinical equivalent doses had the best effect, the 15-fold ZRH intestinal absorption fluid was finally determined as the study subject. Spectral effect relationship showed that 52 corresponding peaks in the ZRH-containing intestinal fluid were positively correlated with the efficacy, including organic acids, phenylpropanoids, iridoids, flavonoids, bile acids, coumarins and chromones. Target validation results showed that 86.9%-96.2% of the total components processed good binding activities with the key targets of PPARα, PPARγ, PPARβ, RXRA and NF-κB, and the docking energy values were all less than -6.0 kcal·mol-1(1 cal≈4.19 J). The results of validation showed that, compared with the normal group, the model group showed a significant increase in the levels of SRA1 and PPARγ mRNA expression, a significant decrease in ABCA1 mRNA expression, a significant increase in the level of PPARγ protein expression, and a significant increase in the levels of IL-1β and NF-κB(P<0.01), compared with the model group, the 15-fold intestinal absorption fluid group showed a significant decrease in the levels of SRA1 and PPARγ mRNA expression(P<0.05, P<0.01), ABCA1 mRNA expression level was significantly up-regulated, the levels of IL-1β and NF-κB were significantly reduced(P<0.01), and PPARγ protein expression level was significantly reduced(P<0.05). ConclusionThis study identifies 52 components and their metabolites in ZRH intestinal absorption fluid that are positively correlated with the inhibition of macrophage foaming, which may be related to the regulation of the PPARs pathway in cells and the reduction of the levels of inflammatory factors, and can provide a reference for the quality control and clinical application of ZRH.
2.Clinicopathological features of radioactive iodine-refractory differentiated thyroid cancer
Beibei JIANG ; Shuai LI ; Wen LIU ; Jingli XUE ; Min LI ; Peifeng LI
Chongqing Medicine 2024;53(1):11-16
Objective To investigate the clinicopathological characteristics of radioactive Iodine-refrac-tory differentiated thyroid cancer(RAIR-DTC)to provide a clinical evidence for early prediction of the thyroid cancer patients with radioactive Iodine-refractory(RAIR).Methods The data of 84 patients with undergoing thyroidectomy and 131I therapy in PLA 960 hospital from January 2010 to December 2019 were retrospectively analyzed.Thirty-nine patients with diagnosed RAIR-DTC served as the study group and 45 cases of radioactive iodine-avid differentiated thyroid cancer(RAIA-DTC)served as the control group.The clinicopathological characteristics were compared between the two groups.The logistic regression was used to analyze the inde-pendent risk factors of RAIR-DTC,and the RAIR-DTC prediction model was established.Results Compared with the RAIA-DTC group,the RAIR-DTC group had more iodine treatment times,the proportions of the pa-tients with age ≥55 years old,total iodine therapeutic dose,distant metastasis,TNM stage Ⅳ,high-risk sub-types and focal calcification were higher,the tumor maximum diameter was greater,the number of lymph node metastases was more and the probability of Ⅱ,Ⅰ+Ⅱ and non-central lymph node metastases was higher(P>0.05).The progression-free survival rate had statistical difference between the two groups(P<0.05).The total survival rate had no statistical difference between the two groups(P>0.05).The binary logistic re-gression analysis results showed that the distant metastasis,high-risk histological subtype and maximum tumor diameter ≥10.5 mm were the independent risk factors for RAIR-DTC.The obtained fitting equation logit(P)=-2.259+3.330X1+2.287X2+1.606X3,the ROC curve was used to calculate the truncation val-ue of the fitted equation as-0.312 5,when logit(P)>-0.312 5,it might develop into RAIR-DTC.Conclusion The clinicopathological characteristics of the patients with differentiated thyroid cancer could ef-fectively predict RAIR.
3.The improvement effect and metabolic mechanism of Echinops sphaerocephalus extract on osteoporosis model rat
Xin DONG ; Jiaqi WANG ; Xiuyan ZHANG ; Zhongyao ZHANG ; Jingkun LU ; Jianping GAO ; Peifeng XUE
China Pharmacy 2023;34(4):413-418
OBJECTIVE To study the intervention effect and metabolic mechanism of Mongolian medicine Echinops sphaerocephalus extract on D-galactose-induced osteoporosis. METHODS Thirty-six 12-week-old male Wistar rats were selected and randomly divided into blank group, model group, Gushukang group, E. sphaerocephalus high-dose, medium-dose and low- dose groups, with 6 rats in each group. Except for blank group, other groups were intraperitoneally injected with D-galactose at 120 mg/kg per day. After 8 weeks of continuous injection, E. sphaerocephalus high-dose, medium-dose and low-dose groups were given drugs intragastrically at dose of 878, 439, 219.5 mg/kg, respectively. Gushukang group was given Gushukang 105.1 mg/kg intragastrically, once a day, for consecutive 8 weeks. After last administration, blood was collected from the abdominal aorta. Enzyme-linked immunosorbent assay was used to measure the contents of bone metabolism indexes [hydroxyproline (HYP), alkaline phosphatase (ALP)] and oxidative stress indexes [total antioxidant capacity (TAOC), superoxide dismutase (SOD), malondialdehyde (MDA)] in serum of rats. Positron emission tomography/computedtomography (PET/CT) was used to analyze the changes of bone microstructure in right tibia bone. Meanwhile, metabolomic technology was used to study the regulation effect of E. sphaerocephalus on osteoporosis model rats. RESULTS Compared with blank group, HYP, ALP, MDA, ratio of bone surface to bone volume (BS/BV), and trabecular separation (Tb·Sp) in model group were significantly increased (P<0.05), while TAOC, SOD, bone mineral density (BMD), bone volume fraction (BVF), trabecular E-mail:Xpfdc153@163.com thickness (Tb·Th) and trabecular number (Tb·N) were significantly decreased (P<0.05). Compared with model group, above indexes of administration groups were all reversed to different extents. The results of metabonomics study showed that after intervened with the extract of E. sphaerocephalus, 18 metabolites such as arachidonic acid, phenylalanine, tyrosine, tryptophan, isoleucine and uric acid in the serum of rats changed significantly, involving 15 metabolic pathways such as arachidonic acid, phenylalanine and tyrosine, of which arachidonic acid metabolism, phenylalanine metabolism and tyrosine metabolism were the main influencing pathways. CONCLUSIONS E. sphaerocephalus extract can effectively improve D-galactose-induced oxidative stress and the deterioration of bone microstructure, which interferes with metabolic pathways such as arachidonic acid metabolism and amino acid metabolism.
4.Absorption Characteristics of Nine Phenylpropanoids in Mongolian Medicine Tabson-2 Decoction in Caco-2 Cells
LI Chunyan ; WANG Xiyue ; LU Jingkun ; DONG Xin ; ZHAO Pengwei ; MA Feixiang ; XUE Peifeng
Chinese Journal of Modern Applied Pharmacy 2023;40(15):2048-2055
OBJECTIVE To study the absorption characteristics of phenylpropanoids of Mongolian medicine Tabson-2 decoction(TBD) in Caco-2 cells and to preliminarily clarify the oral absorption mechanism of TBD. METHODS Caco-2 cell monolayer model was used to analyze the uptake components of TBD in Caco-2 cells by UPLC-MS/MS, and UPLC-MS/MS analysis method was established to determine the nine best absorbed components of TBD, protocatechuic acid, neochlorogenic acid, chlorogenic acid, cryptogenic acid, 1,5-dicaffeinate quinic acid, isochlorogenic acid C, caffeic acid, dihydrocaffeic acid, chlorogenic acid. The effects of time, concentration and P-glycoprotein inhibitor on the absorption of each component were investigated. RESULTS The overall intake of caffeic acid and dihydrocaffeic acid showed an upward trend in 0-180 min, and did not show saturation. The absorption of 3-hydroxycinnamic acid was constant at about 90 min and tended to saturation. The intakes of cryptochlorogenic acid, 1,5-dicaffeinate, quinic acid, isochlorogenic acid C, neochlorogenic acid, chlorogenic acid and protocatechuic acid first decreased and then increased with time from about 90 min. The addition of P-glycoprotein inhibitor verapamil and cyclosporin A had an effect on the absorption of dihydrocaffeic acid compared with the phenylpropanoid components, indicated that dihydrocaffeic acid was the substrate of P-glycoprotein. CONCLUSION The main phenylpropanoids of TBD enter Caco-2 mainly by passive diffusion, supplemented by active transport, and the absorption process of the other eight components is not affected by the efflux of P-glycoprotein except dihydrocaffeic acid.
5.Optimization of the extraction technology of modified Tabusen- 2 and its inhibitory effects on osteoclast differen- tiation
Shu GUO ; Qingxiang SONG ; Rui DONG ; Qianxi FENG ; Peifeng XUE ; Xin DONG
China Pharmacy 2022;33(16):1943-1949
OBJECTIVE To optimize the extraction technology of modified Tabusen- 2(MT-2),and to investigate inhibitory effects of the extract obtained by the optimal technology on osteoclast differentiation. METHODS The index components of MT- 2 process optimization were selected by using network pharmacology. Based on single factor tests ,the extraction technology of MT- 2 was optimized by Box-Behnken design-response surface methodology according to the comprehensive score of contents of above index components ,and then validated. RAW 264.7 cells were induced by receptor activator of nuclear factor-κB ligand(100 ng/mL) to prepare osteoclast differentiation model. Inhibitory effects of MT- 2 extract(18.6,37.2,74.4 ng/mL)obtained by the optimal technology on osteoclast differentiation were investigated. RESULTS The index components screened by network pharmacology included chlorogenic acid ,terpineol diglucoside ,isochlorogenic acid A ,1,5-dicaffeoylquinic acid ,hydroxysafflower yellow A , ginsenoside Rg 1 and ginsenoside Rb 1. The optimal extraction technology of MT- 2 was ethanol volume fraction of 60% ,the solid-liquid ratio of 1 ∶ 14(g/mL),extraction time of 94 min and extraction times of twice. The average comprehensive score obtained by the three validation experiments was 95.50,and the relative error with the predicted value (95.75)was -0.26%. Compared with osteoclastic differentiation model cells ,the cells treated with MT- 2 extract prepared by the optimal technology were mostly mononuclear round cells ,and the number of osteoclasts decreased significantly (P<0.05),its inhibitory effects tended to strengthen with the increase of drug concentration. CONCLUSIONS The optimal extraction technology of MT- 2 is stable and feasible. Obtained extract can inhibit osteoclast differentiation.
6.Simultaneous Determination of Liquiritin and Glycyrrhizic Acid in Wuwei Shaji Granules by HPLC
Jie GENG ; Lihua CONG ; Jinghui FAN ; Chunyu ZHAO ; Peifeng XUE
China Pharmacy 2017;28(27):3836-3838
OBJECTIVE:To establish a method for simultaneous determination of liquiritin and glycyrrhizic acid in Wuwei shaji granules.METHODS:HPLC method was adopted.The determination was performed on Thermo scientific ODS-2 Hypersil column with mobile phase consisted of acetonitrile-0.5% phosphoric acid (gradient elution) with flow rate of 1.0 mL/min.The detection wavelength was set at 237 nm,and column temperature was 25 ℃.The sample size was 20 μL.RESULTS:The linear ranges of liquiritin and glycyrrhizic acid were 48-480 μg(r=0.999 9) and 90-900 μg(r=0.999 9),respectively.The limits of quantitation were 0.55,2.10 μg/mL,and the limits of detection were 0.16,0.62 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2.0%.The recoveries were 95.08%-97.58% (RSD=0.93%,n=6)and 95.86%-99.89% (RSD=1.67%,n=6),respectively.CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the content determination of liquiritin and glycyrrhizic acid in Wuwei shaji granules.
7.A comparative study of four methods of establishing orthotopic human renal cell carcinoma models in nude mice
Peipei ZHAO ; Xiaojing CHEN ; Qiaoling WANG ; Xue ZHAO ; Yanan ZHAO ; Peifeng LIU ; Huili DAI
China Oncology 2017;27(3):177-185
Background and purpose: Renal cell carcinoma is the most common form of kidney cancer, characterized by lack of early symptoms and high malignancy. This study aimed to establish orthotopic nude mice models of human renal cell carcinoma with high success rate and good repeatability. Methods: The four types of methods which were adopted to establish the orthotopic models of renal cell carcinoma were orthotopic injection of 786-0 and ACHN cell suspensions, orthotopic injection of primary cell suspensions obtained from the subcutaneous tumor tissues, renal subcutis orthotopic implantation into renal capsule and surgical subcutis orthotopic implantation into renal fascia. To gain insights into the tumorigenicity and the growth of transplantation tumors, the imageological examination (PET/CT), histological examination (H-E staining, immunohistochemistry staining) and biochemical analysis of blood were carried out. Results: In terms of the subcutaneous transplantation of human renal cell carcinoma models in nude mice, tumorigenic rate of ACHN cells (90%) was higher than that of 786-0 cells (30%). The tumorigenic incidences of 786-0 cell suspensions orthotopic injection, ACHN cell suspensions orthotopic injection, ACHN subcutis cellular suspensions orthotopic injection, ACHN subcutis orthotopic implantation into renal capsule and renal fascia were 33%, 80%, 90%, 100% and 20%, respectively. ACHN subcutis orthotopic implantation into renal capsule was the most effective approach. Imageological and histological results accorded with poorly differentiated renal cell carcinoma. Conclusion: Four orthotopic nude mice models of human renal cell carcinoma were successfully established. Among these methods, ACHN subcutis orthotopic implantation into renal capsule is the most effective approach, which provides an ideal model for the research on biological behavior of human renal cell carcinoma and its treatment.
8.Research Progress in Medicinal Plant Echinops Latifolius Tausch
Dongyan ZHANG ; Lei YANG ; Peifeng XUE ; Jianping GAO
China Pharmacist 2015;(11):1966-1971
Echinops latifolius Tausch is a plant of compositae echinops, whose dried roots are used as traditional Chinese medicine Yuzhou Loulu, and dried inflorescence is used as Mongolian medicine LanCitou. The studies on Chinese medicine Yuzhou Loulu and Mongolian medicine LanCitou in recent years were systemized and compared with each other in the paper, including chemical composi-tion, process research, quality control, pharmacological effects and so on. The results can provide scientific basis for the further studies and utilization of Echinops latifolius Tausch.
9.Need-based design of medical mobile learning platform
Juanping WU ; Wenjie LI ; Xiaoyu WANG ; Wen ZHANG ; Jia XUE ; Peifeng HE ; Jianguang WU
Chinese Journal of Medical Library and Information Science 2014;(2):68-71
The medical mobile learning platform was constructed according to the information need of teachers and students in Shanxi Medical University, Changzhi Medical College, and Fenyang Medical College.The teaching and learning resources in Shanxi Medical University were integrated into the platform which could thus provide a variety of interactive learning ways for its users and users could rapidly find out their interested information resources. However, the platform construction needs the implementation of incentive measures, and regulations and rules for the protection of intellectual property rights.
10.Chemical constituents from Myricaria alopecuroides.
Zhanjun LI ; Peifeng XUE ; Hongxia XIE ; Xiaojuan LI ; Min XIE
China Journal of Chinese Materia Medica 2010;35(7):865-868
OBJECTIVETo investigate the chemical constituents in the leaves and branches of Myricaria alopecuroides.
METHODSolvent extraction method was employed to extract and partition. The chemical constituents were isolated by column chromatography on silica gel, Sephadex LH-20, highly porous resin HP-20. The structures of the compounds were elucidated on the basis of physiochemical properties and spectral analysis.
RESULTEleven compounds were isolated from this plant and identified as ellagic acid 3,3',4-trimethylether (1), ellagic acid 3,3'-dimethylether (2), isorhamnetin (3), kaempferol (4), 3, 5-dihydroxy-4-methoxybenzoic acid (5), daucosterol (6), 6,7,10-trihydroxy-8-octadecenoic acid (7), quercetin (8), gallic acid (9), palmitic acid (10), hexadecanoic acid, 2,3-dihydroxypropyl ester (11).
CONCLUSIONExcept 8 and 9, all compounds were isolated from M. alopecuroides for the first time. Compound 1, 2, 5, 7, 10, 11 were obtained from the genus Myricaria for the frist time.
Organic Chemicals ; analysis ; isolation & purification ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Tamaricaceae ; chemistry


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