1.Characteristic ion Identification of Different Original Haliotidis Concha and Its Counterfeits
Xiaojie LIANG ; Guowei LI ; Lin ZHOU ; Qiping HU ; Muxiang LUO ; Jiehao TANG ; Xiangdong CHEN ; Liye PAN ; Dongmei SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):263-269
ObjectiveTo establish a method for the identification of Haliotidis Concha and its counterfeits, and to improve its quality evaluation method. MethodsA total of 17 batches of Haliotis discus hannai, 4 batches of H. ruber, 3 batches of H. laevigata, 3 batches of H. ovina, 3 batches of H. diversicolor, 3 batches of H. asinina, 3 batches of H. iris were collected. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive-Orbitrap-MS/MS) was used to analyze the hydrolysates of different original Haliotidis Concha and its counterfeits, and the potential characteristic ions of each species were screened by Venn diagram. UPLC-triple quadrupole tandem mass spectrometry(UPLC-QqQ-MS/MS) was used to validate the characteristic ions, and the specific detection method of the characteristic ions was established. ResultsA total of 1 182, 167, 47, 89, 104, 203, 424 potential characteristic ions were screened from H. discus hannai, H. ruber, H. laevigata, H. ovina, H. diversicolor, H. asinina and H. iris, respectively. And 9 characteristic ions were selected. The precision, stability and repeatability of the 9 characteristic ions in the established identification method met the requirements. Different original Haliotidis Concha and its counterfeits could detect their own characteristic ions, including m/z 631.83-886.48(double charge) and m/z 631.83-443.74(double charge) of H. discus hannai, m/z 699.28-232.11(double charge) and m/z 699.28-544.27(double charge) of H. ruber, m/z 535.76-752.37(double charge) and m/z 535.76-548.28(double charge) of H. laevigata, m/z 708.35-442.28(double charge) and m/z 708.35-215.14(double charge) of H. ovina, m/z 561.33-614.86(triple charge), m/z 561.33-468.28(triple charge), m/z 608.29-618.32(double charge) and m/z 608.29-390.21(double charge) of H. diversicolor, m/z 769.85-274.10(double charge), m/z 769.85-532.75(double charge), m/z 827.43-646.36(single charge), m/z 827.43-257.12(single charge) of H. asinina, and m/z 468.24-576.29(double charge) and m/z 468.24-505.26(double charge) of H. iris. ConclusionIn this study, a total of 9 characteristic ions are screened from 6 kinds of original Haliotidis Concha and its counterfeits, and a specific identification method is established, which is helpful to solve the limitations of the existing quality evaluation methods of Haliotidis Concha, and provide a basis for the production, circulation and medication quality.
2.Characteristic ion Identification of Different Original Haliotidis Concha and Its Counterfeits
Xiaojie LIANG ; Guowei LI ; Lin ZHOU ; Qiping HU ; Muxiang LUO ; Jiehao TANG ; Xiangdong CHEN ; Liye PAN ; Dongmei SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):263-269
ObjectiveTo establish a method for the identification of Haliotidis Concha and its counterfeits, and to improve its quality evaluation method. MethodsA total of 17 batches of Haliotis discus hannai, 4 batches of H. ruber, 3 batches of H. laevigata, 3 batches of H. ovina, 3 batches of H. diversicolor, 3 batches of H. asinina, 3 batches of H. iris were collected. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive-Orbitrap-MS/MS) was used to analyze the hydrolysates of different original Haliotidis Concha and its counterfeits, and the potential characteristic ions of each species were screened by Venn diagram. UPLC-triple quadrupole tandem mass spectrometry(UPLC-QqQ-MS/MS) was used to validate the characteristic ions, and the specific detection method of the characteristic ions was established. ResultsA total of 1 182, 167, 47, 89, 104, 203, 424 potential characteristic ions were screened from H. discus hannai, H. ruber, H. laevigata, H. ovina, H. diversicolor, H. asinina and H. iris, respectively. And 9 characteristic ions were selected. The precision, stability and repeatability of the 9 characteristic ions in the established identification method met the requirements. Different original Haliotidis Concha and its counterfeits could detect their own characteristic ions, including m/z 631.83-886.48(double charge) and m/z 631.83-443.74(double charge) of H. discus hannai, m/z 699.28-232.11(double charge) and m/z 699.28-544.27(double charge) of H. ruber, m/z 535.76-752.37(double charge) and m/z 535.76-548.28(double charge) of H. laevigata, m/z 708.35-442.28(double charge) and m/z 708.35-215.14(double charge) of H. ovina, m/z 561.33-614.86(triple charge), m/z 561.33-468.28(triple charge), m/z 608.29-618.32(double charge) and m/z 608.29-390.21(double charge) of H. diversicolor, m/z 769.85-274.10(double charge), m/z 769.85-532.75(double charge), m/z 827.43-646.36(single charge), m/z 827.43-257.12(single charge) of H. asinina, and m/z 468.24-576.29(double charge) and m/z 468.24-505.26(double charge) of H. iris. ConclusionIn this study, a total of 9 characteristic ions are screened from 6 kinds of original Haliotidis Concha and its counterfeits, and a specific identification method is established, which is helpful to solve the limitations of the existing quality evaluation methods of Haliotidis Concha, and provide a basis for the production, circulation and medication quality.
3.Study on the quality of Bolbostemmatis Rhizoma standard decoction by HPLC fingerprint combined with quantitative analysis of multi-components by single marker
Zhenyu LI ; Xiangyuan ZHOU ; Yifei MA ; Xian QIU ; Minyou HE ; Xiangdong CHEN ; Dongmei SUN
International Journal of Traditional Chinese Medicine 2024;46(1):76-83
Objective:To establish the HPLC fingerprint of Bolbostemmatis Rhizoma standard decoction; To determine the three effective components with similar structure by quantitative analysis of multi-components by single marker (QAMS); To evaluate the quality of Bolbostemmatis Rhizoma standard decoction.Methods:HPLC was adopted to establish the fingerprints of 15 batches of Bolbostemmatis Rhizoma standard decoction. The Chromatographic column was Waters XBridge Phenyl (4.6 mm×250 mm, 5 μm). The mobile phase was acetonitrile-0.1% phosphoric acid solution with gradient elution. Cluster analysis (HCA) and principal component analysis (PCA) were conducted based on the relative peak area of common peaks. The same method as the fingerprint was used to establish QAMS of tubeimoside A, B, C on Bolbostemmatis Rhizoma standard decoction.Results:There were 14 common peaks in the fingerprint of Bolbostemmatis Rhizoma standard decoction. It was confirmed that the peak 3 was L-tryptophan, the peak 11 was tubeimoside B, the peak 12 was tubeimoside C, and the peak 13 was tubeimoside A. 15 batches of Bolbostemmatis Rhizoma standard decoction from different origins were divided into 3 categories by HCA and PCA. There was no significant difference between QAMS and the external standard method (ESM) through the system suitability inspection. Conclusion:This method is accurate, reliable and has good specificity, which can effectively evaluate the quality of Bolbostemmatis Rhizoma standard decoction.
4.Study on quality evaluation of Buddleja officinalis from different habitats based on HPLC fingerprint and content determination
Cuijie WEI ; Xiaoxia LIU ; Zhiwen DUAN ; Yongwei FENG ; Xiaozhou JIA ; Yueyi LIANG ; Xiangdong CHEN ; Dongmei SUN ; Zhenyu LI
International Journal of Traditional Chinese Medicine 2024;46(2):215-221
Objective:To establish UPLC fingerprint method and 2 contents determination methods of Buddleja officinalis; To provide a reference for improving the quality control standard and evaluation of Buddleja officinalis from different habitats.Methods:UPLC method was used to establish the fingerprints of 17 batches of Buddleja officinalis. The similarity evaluation, clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to compare the quality differences of Buddleja officinalis from different habitats. The contents of acteoside and linarin in Buddleja officinalis were determined.Results:There were 12 common peaks in UPLC fingerprints of Buddleja officinalis, six of which were identified as echinacoside, acteoside, cynaroside, isoacteoside, linarin, and apigenin. The fingerprint similarity of 17 batches of Buddleja officinalis was more than 0.9; Buddleja officinalis from different habitats were classified into 2 groups. Five differential markers were determined by OPLS-DA analysis. The order of significance was acteoside > peak 3 > echinacoside > isoacteoside > linarin. Edgeworthia chrysantha was identified by the method of fingerprint as counterfeit. The results of content determination showed that the content of Buddleja officinalis in Hubei and Sichuan was the high and stable.Conclusion:The method can effectively analyze the differences of Buddleja officinalis from different habitats, and provide reference for the quality control of Buddleja officinalis.
5.Evaluation of Mechanical Properties of Three-Dimensional-Printed Metal Vertebral Body Substitutes
Enchun DONG ; Jianfeng KANG ; Changning SUN ; Dichen LI ; Yang LUO ; Ling WANG ; Xiangdong LI
Journal of Medical Biomechanics 2024;39(1):76-83
Objective To study the mechanical properties of titanium mesh and three-dimensional(3D)-printed metal vertebral body substitutes(VBS)to provide guidance for the selection and structural optimization of artificial vertebral implants in clinical practice.Methods The equivalent elastic modulus,equivalent yield strength,and structural failure mode of titanium mesh and 3D-printed porous,truss,and topologically optimized VBS were systematically investigated using compression tests.Results The elastic modulus of the titanium mesh(2 908.73±287.39 MPa)was only lower than that of the topologically optimized VBS.However,their structural strengths and stabilities were inadequate.The yield strength of the titanium mesh(46.61±4.85 MPa)was only higher than that of the porous VBS and it was the first to yield during compression.The porous VBS was insufficient for use as the vertebral implant owing to its poor mechanical strength(18.14±0.17 MPa-25.79±0.40 MPa).The truss VBS had good elastic modulus(2 477.86±55.19 MPa-2 620.08±194.36 MPa)and strength(77.61±0.50 MPa-88.42±1.07 MPa).However,the structural stability of the truss VBS was insufficient,and instability occurred easily during compression.The topologically optimized VBS had the highest elastic modulus(3 746.28±183.80 MPa)and yield strength(177.43±3.82 MPa)among all the tested VBS types,which could provide improved security and stability for artificial vertebral implant in vivo services.Conclusions Topology optimization results in a high strength and high stability VBS design.Moreover,it provides a large design space and great safety margin to provide increased possibilities for lightweight and new material design of future artificial vertebral implants.
6.Establishment of UPLC characteristic chromatogram of Pulsatilla chinensis and its application in origin differentiation and counterfeit identification
Guangming HE ; Rui LUO ; Heping ZENG ; Xiaoying LU ; Xiaolong YANG ; Weisheng LYU ; Yueyi LIANG ; Zhenyu LI ; Dongmei SUN ; Xiangdong CHEN
International Journal of Traditional Chinese Medicine 2024;46(6):743-749
Objective:To establish ultra performance liquid chromatography (UPLC) characteristic chromatogram of Pulsatilla chinensis; To provide reference for the origin identification and quality control of Pulsatilla chinensis. Methods:UPLC Method was adopted. The determination was performed on a column of Agilent SB C18 (2.1 mm×100 mm, 1.8 μm) . The mobile phase was acetonitrile-methanol (2:1) -0.1% phosphoric acid solution by fradient elution at a flow rate of 0.30ml/min. The column temperature was 30 ℃. The detection wavelength was 215 nm. The injection volume was 2 μl. The common counterfeit products and medicinal herbs of Pulsatilla chinensis from different areas were evaluated by comparison of characteristic chromatogram, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Results:There were 9 characteristic peaks in the characteristic chromatogram of Pulsatilla chinensis, and 8 common peaks were identified by high resolution mass spectrometry and comparison of reference materials. Through PCA analysis, it was possible to clearly distinguish the medicinal herbs of Pulsatilla chinensis from different areas. Combined with OPLS-DA analysis, it was found that peak 2, peak 3, peak 6 were the main markers of Pulsatilla chinensis from different producing areas. Conclusion:The established method has good specificity, repeatability and durability, and it can effectively distinguish the common counterfeits of Pulsatilla chinensis, and provide the basis of quality control and selection of origin for Pulsatilla chinensis.
7.Study on UPLC fingerprint of Mume flos at different flowering stages based on chemometrics analysis
Shuang HUANG ; Yueyi LIANG ; Jie YANG ; Weisheng LYU ; Xiaoying LU ; Guangming HE ; Zhipeng CHEN ; Xuxuan HOU ; Tianrui XIA ; Zhenyu LI ; Congyou DENG ; Xiangdong CHEN ; Dongmei SUN
International Journal of Traditional Chinese Medicine 2024;46(7):898-904
Objective:To establish the ultra high performance liquid chromatography (UPLC) fingerprints of Mume flos at different flowering stages; To provide reference for the quality research of Mume flos.Methods:The fingerprints of Mume flos were established by UPLC method, and the common peaks were identified by high performance liquid chromatography high resolution mass spectrometry (LC-MS). Chemometrics analysis was carried out with the fingerprints' common peak area of plum blossom at different flowering stages as a variable. Semiquantitative analysis of changes in flavonoids and phenolic acids in Mume flos at different flowering stages was conduct using peak area calculation method.Results:Totally 31 common peaks were identified in the fingerprints of plum blossom medicinal materials at different flowering stages and 9 components were identified. Clustering analysis (HCA) and principal component analysis (PCA) both classified plum blossom medicinal herbs at different flowering stages into three categories. Among them, there were significant differences between the groups at the bud stage, blooming period, and final flowering period, while the differences between the groups at blooming period and final flowering period were relatively small. The orthogonal partial least squares discriminant analysis (OPLS-DA) screened 16 different components with VIP>1.0. The contents of phenolic acids in different flowering stages were as follows: bud stage>blooming period>final flowering period, while the contents of flavonoids were as follows: blooming period>final flowering period>bud stage.Conclusions:This method is simple and reliable, and can provide reference for the quality evaluation of plum blossom medicinal materials at different flowering stages.
8.A Multicenter, Randomized, Double-blind, and Placebo-parallel Controlled Trial of Tibetan Medicine Ruyi Zhenbaowan in Treatment of Knee Osteoarthritis
Chunquan SUN ; Yanming XIE ; Jinghua GAO ; Weiheng CHEN ; Lianxin WANG ; Shangquan WANG ; Xiangdong TIAN ; Zujian XU ; Yuxin ZHENG ; Mingwang ZHOU ; Chungen LI ; Zhanwang XU ; Jiayi GUO ; Shuangqing DU ; Qigang CHEN ; Quan JI ; Zhiqiang BAI ; Jing XIAO ; Wanli QI ; Weiyi YANG ; Jingxiao ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):57-67
ObjectiveThis study aimed to evaluate the clinical efficacy of Ruyi Zhenbaowan(RYZBW)in the treatment of initial and early knee osteoarthritis (KOA) through a prospective multicenter,randomized,double-blind,and placebo-parallel controlled trial. MethodFrom October 13th, 2021 to December 25th, 2021, 240 KOA subjects meeting the acceptance criteria were enrolled in 15 sub-centers including Wangjing Hospital, Chinese Academy of Chinese Medical Sciences, and they were randomly divided into observation group and control group, with 120 cases in each group. The intervention measures for the observation group were RYZBW + health education, and the intervention measures for the control group were RYZBW placebo + health education. The intervention period in both groups was four weeks, and they were followed up for four weeks after the intervention. The primary outcome measure was the total score of Western Ontario and McMaster University Osteoarthritis Index score (WOMAC score), and the secondary outcome measures were the response rate of visual scale (VAS) pain score, WOMAC sub item scores (joint pain, joint stiffness, and joint function), quality of life (SF-12) score, and traditional Chinese medicine (TCM) syndrome score. Result(1) Efficacy evaluation. The marginal model results showed that the observation group was better than the control group in improving the WOMAC total score and WOMAC pain score in the treatment of KOA with RYZBW, and the difference was statistically significant (P<0.05). There was no significant difference between the two groups in improving VAS score response rate, WOMAC function score, WOMAC stiffness score, SF12-PCS (quality of life-physical health) score, SF12-MCS (quality of life-mental health) score, and TCM syndrome score. (2) Subgroup analysis. ① In terms of VAS score response rate, the response rate of the observation group was higher than that of the control group for subjects with baseline VAS score of (4, 5], and the difference was statistically significant (P<0.05). ② In terms of TCM syndrome score, for subjects aged [56, 60] and [61, 65], the decrease in total TCM syndrome score in the observation group was better than that in the control group, and the difference was statistically significant (P<0.05). ConclusionTibetan medicine RYZBW has good clinical efficacy in improving WOMAC total score, VAS score response rate, WOMAC pain score, WOMAC function score, and TCM syndrome score for patients with initial and early KOA, which can fill the lack of Tibetan medicine RYZBW in the treatment of KOA and make a demonstration study for the inheritance and development of ethnic medicine.
9.Study on the quality markers of Curcumae Radix standard decoction based on fingerprint and network pharmacology
Zhiwen DUAN ; Xiaoxia LIU ; Minyou HE ; Cuijie WEI ; Yongwei FENG ; Haibao QIU ; Ronghui ZHENG ; Dongmei SUN ; Xiangdong CHEN ; Zhenyu LI
International Journal of Traditional Chinese Medicine 2024;46(12):1622-1628
Objective:To establish the ultra-high performance liquid chromatography (UPLC) fingerprint and high performance liquid chromatography (HPLC) content determination method of Curcumae Radix standard decoction; To predict the quality markers of Curcumae Radix standard decoction combined with network pharmacology.Methods:UPLC method was used to establish the fingerprint of Curcumae Radix standard decoction, and the common peaks were determined. Combined with chemical pattern recognition techniques such as similarity analysis and clustering analysis, Curcumae Radix standard decoction from different producing areas was studied, and curcumol was used as an index to determine the content of 24 batches of Curcumae Radix standard decoction. At the same time, network pharmacology was used to predict potential of curcumol and (1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β-(3-oxobutyl) bicyclo [4.1.0] heptan-3-one.Results:A total of 24 batches of Curcumae Radix standard decoction from different habitats were compared and analyzed, and 10 common peaks were calibrated. The similarity of 24 batches of samples ranged from 0.982 to 0.999. Clustering analysis and principal component analysis divided them into three categories. Heat map analysis showed that peak 8 (curcumol) and peak 9 ((1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β-(3-oxobutyl) bicyclo [4.1.0] heptan-3-one) were the main components. The content of curcumol in 24 batches of Curcumae Radix standard decoction was 0.69-1.87 mg/g; curcumol and (1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β- (3-oxobutyl) bicyclo [4.1.0] heptan-3-one may regulate the neuroactive ligand-receptor interaction signaling pathway, calcium signaling, and excitation by regulating neuroactive ligand-receptor interaction signaling pathway, calcium signaling, and excitation. It was preliminarily predicted that curcumol and (1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β-(3-oxobutyl) bicyclo [4.1.0] heptan-3-one were potential quality markers of Curcumae Radix.Conclusion:Curcumol and (1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β-(3-oxobutyl) bicyclo [4.1.0] heptan-3-one are potential quality markers of Curcumae Radix standard decoction, and the established fingerprint can be used for the quality control of Curcumae Radix standard decoction.
10.Effects of radiation on brain based on EEG signal
Chengyu HE ; Jundong FENG ; Xiangdong SUN ; Liuxin TIAN ; Hongyu BO ; Simin SUN ; Hao FU ; Qian LI
Space Medicine & Medical Engineering 2024;35(3):149-155
Objective Space radiation is an important environmental factor affecting the health and efficiency of astronauts.To explore the effects of radiation on electroencephalogram(EEG)signal,and to provide references for further study of the effects of radiation on Central Nervous System(brain)injury.Methods The resting state EEG of radiotherapy volunteers were collected before and after X-ray exposure.The effects of radiation on EEG was studied by means of spectral analysis,nonlinear dynamic characteristic analysis,correlation coefficient analysis and microstate analysis.Results The results show that radiation induced a"slow wave"phenomenon in the EEG spectrum,with a decrease in Gravity Frequency,Sample Entropy,and Lempel-Ziv Complexity,and an increase in Fatigue Index.The average duration and frequency of EEG microstate C both significantly decreased,and the proportion of time for microstate D decreased,while the average correlation coefficient between microstates A and C increased.Conclusion A decrease was induced by radiation in neuronal excitability,which may affect cognitive brain networks and brain attention.This study can provide reference for the study of radiation damage to the Central Nervous System(brain).

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