1.In Vitro and In Vivo Chemical Composition Analysis of Reference Sample of Jinshui Liujunjian Based on UPLC-Q-TOF-MS/MS
Xinyue YANG ; Huiyu LI ; Yaqi LOU ; Xingxing WANG ; Guifang YU ; Chenfeng ZHANG ; Zhenzhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):166-173
ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was used to detect the reference sample solution, plasma, and tissue samples of Jinshui Liujunjian under positive and negative ion modes, respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times, secondary mass spectral fragments, reference standards, and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian, including 47 flavonoids, 5 amino acids, 13 iridoids, 16 triterpenoid saponins, etc., of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components; 50 prototype components were identified in different tissues, among which 13, 10, 7, 21, 11, 6, 14, and 40 prototype components were identified in the heart, liver, spleen, lung, kidney, brain, large intestine, and stomach, respectively. Among them, 7 compounds such as ferulic acid, glycyrrhizic acid, and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian, primarily composed of flavonoids and triterpenoid saponins, along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.
2.In Vitro and In Vivo Chemical Composition Analysis of Reference Sample of Jinshui Liujunjian Based on UPLC-Q-TOF-MS/MS
Xinyue YANG ; Huiyu LI ; Yaqi LOU ; Xingxing WANG ; Guifang YU ; Chenfeng ZHANG ; Zhenzhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):166-173
ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was used to detect the reference sample solution, plasma, and tissue samples of Jinshui Liujunjian under positive and negative ion modes, respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times, secondary mass spectral fragments, reference standards, and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian, including 47 flavonoids, 5 amino acids, 13 iridoids, 16 triterpenoid saponins, etc., of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components; 50 prototype components were identified in different tissues, among which 13, 10, 7, 21, 11, 6, 14, and 40 prototype components were identified in the heart, liver, spleen, lung, kidney, brain, large intestine, and stomach, respectively. Among them, 7 compounds such as ferulic acid, glycyrrhizic acid, and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian, primarily composed of flavonoids and triterpenoid saponins, along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.
3.Mechanism of Congrong Zonggan Capsules in Improving Neuroinflammation and Cognitive Impairment in 5×FAD Mice Based on NF-κB/NLRP3 Signaling Pathway
Yanru ZHOU ; Xinru GU ; Yuru LIU ; Shun ZHANG ; Yaozhong LYU ; Zhenzhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):130-138
ObjectiveTo investigate the effects of Congrong Zonggan capsules (CRZG) on cognitive impairment in the Alzheimer's disease (AD) model of mice and its related mechanisms. MethodsSPF grade 4-week-old 5×FAD mice were divided into a model group, low-dose CRZG (0.819 g·kg-1) and high-dose CRZG (1.638 g·kg-1) groups, and Donepezilepezil hydrochloride group (2 mg·kg-1), with eight mice in each group. Eight C57 mice with the same background were set as the normal group. After one week of adaptive feeding, mice were orally administered continuously for six months. On the 5th month of drug administration, Y maze, new object recognition, and Morris water maze tests were conducted separately. After administration, mouse brain tissue was taken, and the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in brain tissue were detected by enzyme-linked immunosorbent assay (ELISA). Immunofluorescence (IF) was used to detect the expression of small glial cell markers Iba1, astrocyte markers GFAP, and amyloid protein 1-42 (Aβ1-42) in the hippocampus of the brain tissue. The hematoxylin-eosin (HE) staining was used to detect pathological changes in the hippocampus of brain tissue. Western blot was used to detect the expression of nuclear factor-κB (NF-κB) p65, NOD-like receptor protein 3 (NLRP3), cleaved Caspase-1, apoptosis-associated speck-like protein containing a CARD (ASC), and other proteins in the brain tissue. ResultsCompared with those in the normal group, the mice in the model group had obvious cognitive impairment. The spontaneous alternation rate of the Y maze was decreased, and the discrimination index of novel object recognition was decreased significantly (P<0.01). The escape latency in the water maze was shortened significantly (P<0.01). The contents of IL-6 and TNF-α in brain tissue were increased. The fluorescence levels of Iba1 and Aβ1-42 in the hippocampus were significantly increased (P<0.01). There was a significant increase in neuronal lesions, neuronal atrophy, loose arrangement of tissue structure, and abnormal erythrocyte aggregation in the hippocampus. The protein expressions of p-NF-κB p65/NF-κB p65, cleaved Caspase-1, ASC, IL-6, and IL-1β were significantly increased (P<0.05, P<0.01). Compared with the model group, the spontaneous alternation rate and discrimination index of the high-dose CRZG group were increased significantly (P<0.01), and the escape latency was shortened significantly (P<0.05, P<0.01). The content of IL-6 decreased in the brain, and that of TNF-α dropped significantly (P<0.01). The expression of Iba1 protein and Aβ1-42 in the hippocampus decreased significantly (P<0.05, P<0.01). The hippocampal neurons were densely arranged, and the pyramidal nuclei were clear and centered. The abnormal aggregation of red blood cells was alleviated. The value of p-NF-κB/NF-κB proteins and the expression of ASC, cleaved Caspase-1, IL-6, and IL-1β were significantly decreased (P<0.05, P<0.01). ConclusionCRZG can effectively improve cognitive impairment in 5×FAD mice with Alzheimer's disease, and its mechanism may be related to the regulation of the NF-κB/NLRP3 pathway to reduce the abnormal activation of microglia and inhibit neuroinflammation.
4.Exploration of the Pharmacological Substances Basis and Potential Mechanism of Anchang Formulation in the Treatment of Ulcerative Colitis Based on UPLC-Q-TOF-MS Technology and Network Pharmacology
Mingxia WU ; Nan WANG ; Yelin DING ; Mengsitong LI ; Yunqi CUI ; Zhenzhong WANG ; Yingbo YANG ; Wei XIAO
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(5):622-636
OBJECTIVE To identify and characterize the chemical ingredients of Anchang formulation,further screen the active ingredients of this formulation treating ulcerative colitis by network pharmacology,and explore the potential targets and pathways,provi-ding scientific basis for its mechanism research and clinical application.METHODS Chemical ingredients in Anchang formulation were acquired by Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS)technology and literature retrieval.The potential active ingredients and key targets for the treatment were obtained from Swiss Target Prediction,GeneCards,STRING,and then Gene Ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were analyzed in the DAVID database.The interactions between the active ingredients and the core targets were verified by using the AutoDock software.The RAW 264.7 murine-derived macrophage inflammation model was also established to val-idate the anti-inflammatory activity of the pre-screened chemical ingredients and further explore the related mechanisms.RESULTS In this study,108 chemical ingredients of Anchang formulation were characterized by UPLC-Q-TOF-MS technology,and expanded to 134 through literature search.The component-target network where 39 core active components were screened was further constructed,and 15 key therapeutic targets were screened by the protein-protein interaction network constructed.The enrichment analysis of KEGG pathway indicated that Anchang formulation can regulate TNF,PI3K-Akt,MAPK,cancer and other related signaling pathways and ex-ert a therapeutic effect.The results of cell experiments showed that Anchang formulation and its active ingredients could inhibit the re-lease of NO,TNF-α and IL-6 in the LPS-induced RAW 264.7 cell inflammation model.CONCLUSION Based on the concept of"ingredient-target-pathway",this study evaluates the anti-inflammatory effect of Anchang formulation and its active ingredients,pre-dicts the potential mechanism of treatment for UC,and provides a theoretical basis and research ideas for the quality control of the for-mulation and its treatment for UC.
5.Progress in the Study of the Chemical Composition and Biological Activity of Hypericum Attenuatum Choisy
Xiling FAN ; Wenjun LIU ; Xueni NIU ; Liang CAO ; Jinzhou TIAN ; Xin WANG ; Zhenzhong WANG ; Wei XIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(6):1578-1591
Hypericum attenuatum Choisy.is dry whole grass of the genus Hypericum L.,is a kind of commonly used folk medicinal herbs more than 2400 years.And it is often used to treat heart disease,hemostasis,scald.Based on a review of domestic and international literature,the main chemical components of Hypericum attenuatum Choisy.include PPAPs,flavonoids,and volatile oil,of which PPAPs and xanthone have received the attention of a large number of scholars because of their complex and novel structures and unique pharmacological effects.Modern pharmacological studies have shown that Hypericum attenuatum Choisy.exerts various pharmacological activities,including anti-arrhythmia,reducing blood sugar,anti-tumor,anti-virus,anti-inflammation,as well as the treatment of depression.As a valuable folk medicine,there is relatively little related traditional Chinese medicine products,this review focus on its phytochemistry,and pharmacology,providing a comprehensive perspective and novel ideas for exploring its current and potential applications.
6.The expression of LAMB3 and ITGB4 in esophageal squamous cell carcinoma and their relationship with clinicopathological features
Yali ZHAO ; Sheng WU ; Wei PENG ; Wanxiang WANG ; Haoran ZHANG ; Zhenzhong FENG ; Li MA ; Xian WANG
Chinese Journal of Clinical and Experimental Pathology 2025;41(10):1301-1307,1313
Purpose To investigate the expression of LAMB3 and ITGB4 in esophageal squamous cell carcinoma(ESCC)and analyze their associations with clinicopathological features.Methods Bioinformatics was used to assess the expression of LAMB3 and ITGB4 in pan-cancer and ESCC.Immunohistochemistry was performed to evaluate their expression and distribution in ESCC tissues,and correlations clinicopathological features were analyzed.Follow-up data were collected to construct Kaplan-Meier survival curves,and Cox regression analysis was proformed to determine their prognostic significance.Results Bioinformatics analysis showed that LAMB3 and ITGB4 were highly expressed in ES-CC tissues(P<0.001).Immunohistochemistry confirmed that both proteins were localized in the cytoplasm and membrane of tumor cells.High LAMB3 expression was significantly associated with poor differentiation(P<0.001),lymph node metastasis(P=0.015),and T staging(P<0.001).High ITGB4 expression was significantly correlated with poor differentiation(P=0.004)and advanced T staging(P=0.004).Cox regression analysis identified high ex-pression of LAMB3 and ITGB4 as independent prognostic factors for overall survival[HR=4.97(95%CI:2.73-9.02);HR=2.33(95%CI:1.36-3.99)].Conclusion LAMB3 and ITGB4 are highly expressed in ESCC.High LAMB3 expression is associated with tumor differentiation,lymph node metastasis,and T staging,while high ITGB4 expression is correlated with tumor differentiation and T staging.Patients with high expression of LAMB3 or ITGB4 have poorer overall survival,suggesting that these proteins may serve as prognostic biomarkers for unfavorable outcome in ESCC.
7.Analysis of Chemical Components of Yin-Qiao-Qing-Re Tablets by UPLC-Q-TOF-MS/MS and GC-MS
Zerong CAI ; Yumei HU ; Wenjun LIU ; Shulin WANG ; Xinyu KONG ; Yifan YANG ; Mengyu QIAN ; Li-ang CAO ; Zhenzhong WANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(9):1198-1212
OBJECTIVE The non-volatile and volatile chemical components in Yin-Qiao-Qing-Re Tablets were analyzed sepa-rately using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS)and Gas Chromatography Mass Spectrometry(GC-MS).METHODS The non-volatile components were analyzed using a Waters ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7 μm),with a mobile phase consisting of 0.1%formic acid aqueous solution(A)and acetonitrile(B)for gradient elution,a flow rate of 0.35 mL·min-1,an injection volume of 5 μL,and a column temperature of 30 ℃;the volatile components were analyzed using an Agilent SH-I-5MS column(5%Phenyl Methyl Silox,30 m×250 μm,0.25 μm);the procedure was temperature-programmed,with an injection volume of 1 μL,a split ratio of 10∶1,a flow rate of 1.0 mL·min-1,and an inlet temperature of 200 ℃.RESULTS A total of 134 non-volatile chemical components and 23 volatile components were analyzed and identified from Yin-Qiao-Qing-Re Tablets,among which 49 compounds were confirmed through comparison with reference stand-ards.The non-volatile components mainly include 27 flavonoids,21 organic acids,15 lignans,14 iridoids,12 phenylethanoid glyco-sides,11 saponins,10 alkaloids,5 terpenes,4 amino acids,3 phenylpropanoids,3 nucleosides,3 xanthones,3 phenolic glycosides,2 chromones and 1 carbohydrate.The volatile components mainly include 11 monoterpenes,5 alcohols and phenols,3 alkenes,2 ke-tones,1 ester,and 1 hydrocarbon.CONCLUSION This study rapidly identifies the chemical components of Yin-Qiao-Qing-Re Tablets,laying a preliminary foundation for research on the pharmacodynamic substances of Yin-Qiao-Qing-Re Tablets and the im-provement of quality control standards.
8.Optimization of the Molding Process of Classical Prescription Xiebai Granules Based on Box-Behnken Design-Response Sur-face Method and BP Neural Network Method
Jingjing WANG ; Zhongkun XU ; Juan FU ; Weili LIU ; Zhaodong HU ; Chenfeng ZHANG ; Zhenzhong WANG ; Wei XIAO
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(10):1333-1343
OBJECTIVE To optimize the molding process of Xiebai Granules(XG)using the Box-Behnken design-response surface method combined with the BP neural network method,and to evaluate the consistency of particle quality between different bat-ches by establishing physical fingerprint.METHODS Dry paste powder was used as the main drug,dry granulation was adopted,and the forming rate,dissolution rate,moisture absorption rate and angle of repose of the granules were used as evaluation indexes,the ex-cipients dextrin,maltodextrin and lactose of the particles,were screened by single factor test combined with simplex-lattice design and entropy weight method,and the optimal excipient ratio was selected.The entropy weight method combined with the Box-Behnken de-sign-response surface method and the BP neural network algorithm were used to optimize the process parameters,and the process veri-fication was carried out.The physical fingerprint was used to comprehensively characterize the bulk density(Da),hygroscopicity(H),moisture(HR),tap density(Dc),angle of repose(α),Hausner ratio(IH),relative uniformity index(Iθ),Carr index(IC),and in-terparticle pore number(Ie),and the consistency of particle quality in different batches was evaluated.RESULTS The optimal ratio of excipients was dextrin 15%,maltodextrin 48%,and lactose 37%.The optimal process parameters were conveying speed 95 r·min-1,pressure wheel speed 4 r·min-1 and hydraulic pressure 7 MPa.The similarity of the physical fingerprints of the five bat-ches of XG was greater than 0.98.CONCLUSION The optimized molding process of XG is stable and feasible,and the quality of different batches of XG is stable,which can provide a reference for the development and industrial scale-up production of XG.
9.Exploration of the Pharmacological Substances Basis and Potential Mechanism of Anchang Formulation in the Treatment of Ulcerative Colitis Based on UPLC-Q-TOF-MS Technology and Network Pharmacology
Mingxia WU ; Nan WANG ; Yelin DING ; Mengsitong LI ; Yunqi CUI ; Zhenzhong WANG ; Yingbo YANG ; Wei XIAO
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(5):622-636
OBJECTIVE To identify and characterize the chemical ingredients of Anchang formulation,further screen the active ingredients of this formulation treating ulcerative colitis by network pharmacology,and explore the potential targets and pathways,provi-ding scientific basis for its mechanism research and clinical application.METHODS Chemical ingredients in Anchang formulation were acquired by Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS)technology and literature retrieval.The potential active ingredients and key targets for the treatment were obtained from Swiss Target Prediction,GeneCards,STRING,and then Gene Ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were analyzed in the DAVID database.The interactions between the active ingredients and the core targets were verified by using the AutoDock software.The RAW 264.7 murine-derived macrophage inflammation model was also established to val-idate the anti-inflammatory activity of the pre-screened chemical ingredients and further explore the related mechanisms.RESULTS In this study,108 chemical ingredients of Anchang formulation were characterized by UPLC-Q-TOF-MS technology,and expanded to 134 through literature search.The component-target network where 39 core active components were screened was further constructed,and 15 key therapeutic targets were screened by the protein-protein interaction network constructed.The enrichment analysis of KEGG pathway indicated that Anchang formulation can regulate TNF,PI3K-Akt,MAPK,cancer and other related signaling pathways and ex-ert a therapeutic effect.The results of cell experiments showed that Anchang formulation and its active ingredients could inhibit the re-lease of NO,TNF-α and IL-6 in the LPS-induced RAW 264.7 cell inflammation model.CONCLUSION Based on the concept of"ingredient-target-pathway",this study evaluates the anti-inflammatory effect of Anchang formulation and its active ingredients,pre-dicts the potential mechanism of treatment for UC,and provides a theoretical basis and research ideas for the quality control of the for-mulation and its treatment for UC.
10.Analysis of Chemical Components of Yin-Qiao-Qing-Re Tablets by UPLC-Q-TOF-MS/MS and GC-MS
Zerong CAI ; Yumei HU ; Wenjun LIU ; Shulin WANG ; Xinyu KONG ; Yifan YANG ; Mengyu QIAN ; Li-ang CAO ; Zhenzhong WANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(9):1198-1212
OBJECTIVE The non-volatile and volatile chemical components in Yin-Qiao-Qing-Re Tablets were analyzed sepa-rately using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS)and Gas Chromatography Mass Spectrometry(GC-MS).METHODS The non-volatile components were analyzed using a Waters ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7 μm),with a mobile phase consisting of 0.1%formic acid aqueous solution(A)and acetonitrile(B)for gradient elution,a flow rate of 0.35 mL·min-1,an injection volume of 5 μL,and a column temperature of 30 ℃;the volatile components were analyzed using an Agilent SH-I-5MS column(5%Phenyl Methyl Silox,30 m×250 μm,0.25 μm);the procedure was temperature-programmed,with an injection volume of 1 μL,a split ratio of 10∶1,a flow rate of 1.0 mL·min-1,and an inlet temperature of 200 ℃.RESULTS A total of 134 non-volatile chemical components and 23 volatile components were analyzed and identified from Yin-Qiao-Qing-Re Tablets,among which 49 compounds were confirmed through comparison with reference stand-ards.The non-volatile components mainly include 27 flavonoids,21 organic acids,15 lignans,14 iridoids,12 phenylethanoid glyco-sides,11 saponins,10 alkaloids,5 terpenes,4 amino acids,3 phenylpropanoids,3 nucleosides,3 xanthones,3 phenolic glycosides,2 chromones and 1 carbohydrate.The volatile components mainly include 11 monoterpenes,5 alcohols and phenols,3 alkenes,2 ke-tones,1 ester,and 1 hydrocarbon.CONCLUSION This study rapidly identifies the chemical components of Yin-Qiao-Qing-Re Tablets,laying a preliminary foundation for research on the pharmacodynamic substances of Yin-Qiao-Qing-Re Tablets and the im-provement of quality control standards.

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