1.Evaluation of Anti-osteoporosis Activity and Hepatotoxicity of Xianling Gubao Based on Zebrafish Model
Qiuman LI ; Yue QIAN ; Zixuan ZHU ; Yuan SONG ; Qian DENG ; Shengyun DAI ; Chongjun ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):87-94
ObjectiveTo investigate the association and translational mechanism between the hepatotoxicity of Xianling Gubao (XLGB) and its treatment of osteoporosis based on a zebrafish model. MethodsZebrafish were randomly selected four days after fertilization (4 dpf) and exposed to different concentrations of XLGB (0.7,0.35 mg·L-1) for 96 h. At the endpoint of the exposure, the mortality rates of zebrafish in the treatment groups of different concentrations were counted, and the "dose-toxicity" curves were plotted. The 10% sublethal concentration (LC10) was calculated. The liver area, acridine orange staining, and pathological tissue sections of transgenic zebrafish [CZ16 (gz15Tg.Tg (fabp 10a: ds Red; ela31: EGFP)] were used as indicators to confirm the hepatic damage caused by the sublethal concentration of XLGB. By using the prednisolone (PNSL)-induced osteoporosis model of zebrafish, the anti-osteoporosis activity of XLGB was evaluated by using the area of skull stained by alizarin red and the cumulative optical density value as indicators. Then, the toxicity difference of XLGB on the liver of zebrafish in healthy and osteoporotic states was compared, and the mechanism of the translational action of the toxicity of XLGB was predicted based on network pharmacology and real-time polymerase chain reaction(Real-time PCR). ResultsThe LC10 of XLGB on zebrafish (8 dpf) was 0.7 mg·L-1. Compared with the blank group, the sublethal concentration (LC10=0.7 mg·L-1, 1/2 LC10=0.35 mg·L-1) of XLGB induced an increase in the number of apoptosis of hepatocytes in a dose-dependent manner, and the tissue arrangement of the liver was disordered and loose. The vacuoles were obvious, and the fluorescence area of the liver was significantly reduced (P<0.01). Compared with the blank group, the mineralized area and cumulative optical density value of zebrafish skull in the PNSL model group were significantly reduced (P<0.01), and those in the 0.7,0.35 mg·L-1 XLGB treatment group were significantly increased compared with the model group (P<0.01). Most importantly, 0.7 mg·L-1 XLGB had no significant effect on the liver of zebrafish in the osteoporosis disease model compared with the blank group. The results of network pharmacology and real-time PCR experiments showed that the toxic transformation of XLGB might be related to the differences in the expression levels of key targets, such as tumor protein 53 (TP53), cysteine aspartic acid specific protease-3(Caspase-3), interleukin(IL)-6, and alkaline phosphatase(ALP) in different organismal states. ConclusionUnder certain conditions, XLGB has hepatotoxicity in normal zebrafish, but under osteoporotic conditions, XLGB not only exerts significant anti-osteoporosis activity but also alleviates hepatotoxicity significantly, which provides a reference for the safe clinical use of XLGB and real evidence for the theories of traditional Chinese medicine of attacking poison with poison and of treating disease with corresponding drugs without damage to the body.
2.The introduction on the revised standards of pharmaceutical excipients in the Chinese Pharmacopoeia 2025 Edition
CHEN Lei ; LUI Yanming ; YUAN Yaozuo ; CHEN Ying ; DAI Hong ; ZHANG Jun ; MA Shuangcheng
Drug Standards of China 2025;26(1):051-057
According to the work goals and tasks determined by edition outline of the Chinese Pharmacopoeia 2025 Edition, the Chinese Pharmacopoeia 2025 has been completed. Among them, 52 new pharmaceutical excipients monographs have been added, and the total number has reached 387. 245 pharmaceutical excipients monographs have been revised, of which 109 monographs have only textual revisions and 136 monographs have substantive revisions. This article focuses on the general framework and the main characteristics of the standards of pharmaceutical excipients in the Chinese Pharmacopoeia 2025, which can contribute to accurately understand and utilize the standards in Chinese Pharmacopoeia.
3.Advantages of modified ligation method for spinal cord injury modeling
Daohui LI ; Xiaoshuang XU ; Zhengtao LI ; Xinpeng TIAN ; Hangchuan BI ; Yuan LIU ; Yongwen DAI ; Lingqiang CHEN
Chinese Journal of Tissue Engineering Research 2025;29(2):379-384
BACKGROUND:Currently,different methods of model establishment have been derived from different injury modes of spinal cord injury.Traditional physical injury modeling methods have their own advantages and disadvantages,and there is a lack of more effective and stable animal models of spinal cord injury. OBJECTIVE:To establish a reproducible,controllable,trauma-free,low-mortality,more stable,widely applicable,and short-term postoperative care rat model of spinal cord injury. METHODS:Forty Sprague-Dawley rats with similar body mass and ages were randomly divided into a control group and an improved group,with 20 rats in each group.Animal models of spinal cord injury in the control group were constructed using a clip model method,while the improved group used a modified ligation method based on the compression method to make the spinal cord injury models using suture ligation based on fenestration.Postoperative comparisons were made between the two groups,assessing urination behavior,hematuria,pyuria(infection rate),mortality,scoliosis rate and Basso-Beattie-Bresnahan locomotor rating scale scores at 1,3,5,and 7 days after modeling. RESULTS AND CONCLUSION:Compared with the conventional modeling method,the modified ligation method based on the compression method resulted in faster recovery of urination behavior,lower hematuria rate,lower infection rate,lower mortality rate,lower scoliosis rate,and more concentrated and stable Basso-Beattie-Bresnahan scores(all below 2 points within 1 week).This proves that the modified ligation method based on compression is more suitable for the establishment of spinal cord injury models in rats.
4.Expression and prognostic value of triggering receptor expressed on myeloid cells-1 in patients with cirrhotic ascites and intra-abdominal infection
Feng WEI ; Xinyan YUE ; Xiling LIU ; Huimin YAN ; Lin LIN ; Tao HUANG ; Yantao PEI ; Shixiang SHAO ; Erhei DAI ; Wenfang YUAN
Journal of Clinical Hepatology 2025;41(5):914-920
ObjectiveTo analyze the expression level of triggering receptor expressed on myeloid cells-1 (TREM-1) in serum and ascites of patients with cirrhotic ascites, and to investigate its correlation with clinical features and inflammatory markers and its role in the diagnosis of infection and prognostic evaluation. MethodsA total of 110 patients with cirrhotic ascites who were hospitalized in The Fifth Hospital of Shijiazhuang from January 2019 to December 2020 were enrolled, and according to the presence or absence of intra-abdominal infection, they were divided into infection group with 72 patients and non-infection group with 38 patients. The patients with infection were further divided into improvement group with 38 patients and non-improvement group with 34 patients. Clinical data and laboratory markers were collected from all patients. Serum and ascites samples were collected, and ELISA was used to measure the level of TREM-1. The independent-samples t test was used for comparison of normally distributed continuous data between two groups; the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups, and the Kruskal-Wallis H test was used for comparison between multiple groups; the chi-square test was used for comparison of categorical data between two groups. A Spearman correlation analysis was used to investigate the correlation between indicators. A multivariate Logistic regression analysis was used to identify the influencing factors for the prognosis of patients with cirrhotic ascites and infection. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic and prognostic efficacy of each indicator, and the Delong test was used for comparison of the area under the ROC curve (AUC). ResultsThe level of TREM-1 in ascites was significantly positively correlated with that in serum (r=0.50, P<0.001). Compared with the improvement group, the non-improvement group had a significantly higher level of TREM-1 in ascites (Z=-2.391, P=0.017) and serum (Z=-2.544, P=0.011), and compared with the non-infection group, the infection group had a significantly higher level of TREM-1 in ascites (Z=-3.420, P<0.001), while there was no significant difference in the level of TREM-1 in serum between the two groups (P>0.05). The level of TREM-1 in serum and ascites were significantly positively correlated with C-reactive protein (CRP), procalcitonin (PCT), white blood cell count, and neutrophil-lymphocyte ratio (r=0.288, 0.344, 0.530, 0.510, 0.534, 0.454, 0.330, and 0.404, all P<0.05). The ROC curve analysis showed that when PCT, CRP, and serum or ascitic TREM-1 were used in combination for the diagnosis of cirrhotic ascites with infection, the AUCs were 0.715 and 0.740, respectively. The multivariate Logistic regression analysis showed that CRP (odds ratio [OR]=1.019, 95% confidence interval [CI]: 1.001 — 1.038, P=0.043) and serum TREM-1 (OR=1.002, 95%CI: 1.000 — 1.003, P=0.016) were independent risk factors for the prognosis of patients with cirrhotic ascites and infection, and the combination of these two indicators had an AUC of 0.728 in predicting poor prognosis. ConclusionThe level of TREM-1 is closely associated with the severity of infection and prognosis in patients with cirrhotic ascites, and combined measurement of TREM-1 and CRP/PCT can improve the diagnostic accuracy of infection and provide support for prognostic evaluation.
5.Network Pharmacology and Experimental Verification Unraveled The Mechanism of Pachymic Acid in The Treatment of Neuroblastoma
Hang LIU ; Yu-Xin ZHU ; Si-Lin GUO ; Xin-Yun PAN ; Yuan-Jie XIE ; Si-Cong LIAO ; Xin-Wen DAI ; Ping SHEN ; Yu-Bo XIAO
Progress in Biochemistry and Biophysics 2025;52(9):2376-2392
ObjectiveTraditional Chinese medicine (TCM) constitutes a valuable cultural heritage and an important source of antitumor compounds. Poria (Poria cocos (Schw.) Wolf), the dried sclerotium of a polyporaceae fungus, was first documented in Shennong’s Classic of Materia Medica and has been used therapeutically and dietarily in China for millennia. Traditionally recognized for its diuretic, spleen-tonifying, and sedative properties, modern pharmacological studies confirm that Poria exhibits antioxidant, anti-inflammatory, antibacterial, and antitumor activities. Pachymic acid (PA; a triterpenoid with the chemical structure 3β-acetyloxy-16α-hydroxy-lanosta-8,24(31)-dien-21-oic acid), isolated from Poria, is a principal bioactive constituent. Emerging evidence indicates PA exerts antitumor effects through multiple mechanisms, though these remain incompletely characterized. Neuroblastoma (NB), a highly malignant pediatric extracranial solid tumor accounting for 15% of childhood cancer deaths, urgently requires safer therapeutics due to the limitations of current treatments. Although PA shows multi-mechanistic antitumor potential, its efficacy against NB remains uncharacterized. This study systematically investigated the potential molecular targets and mechanisms underlying the anti-NB effects of PA by integrating network pharmacology-based target prediction with experimental validation of multi-target interactions through molecular docking, dynamic simulations, and in vitro assays, aimed to establish a novel perspective on PA’s antitumor activity and explore its potential clinical implications for NB treatment by integrating computational predictions with biological assays. MethodsThis study employed network pharmacology to identify potential targets of PA in NB, followed by validation using molecular docking, molecular dynamics (MD) simulations, MM/PBSA free energy analysis, RT-qPCR and Western blot experiments. Network pharmacology analysis included target screening via TCMSP, GeneCards, DisGeNET, SwissTargetPrediction, SuperPred, and PharmMapper. Subsequently, potential targets were predicted by intersecting the results from these databases via Venn analysis. Following target prediction, topological analysis was performed to identify key targets using Cytoscape software. Molecular docking was conducted using AutoDock Vina, with the binding pocket defined based on crystal structures. MD simulations were performed for 100 ns using GROMACS, and RMSD, RMSF, SASA, and hydrogen bonding dynamics were analyzed. MM/PBSA calculations were carried out to estimate the binding free energy of each protein-ligand complex. In vitro validation included RT-qPCR and Western blot, with GAPDH used as an internal control. ResultsThe CCK-8 assay demonstrated a concentration-dependent inhibitory effect of PA on NB cell viability. GO analysis suggested that the anti-NB activity of PA might involve cellular response to chemical stress, vesicle lumen, and protein tyrosine kinase activity. KEGG pathway enrichment analysis suggested that the anti-NB activity of PA might involve the PI3K/AKT, MAPK, and Ras signaling pathways. Molecular docking and MD simulations revealed stable binding interactions between PA and the core target proteins AKT1, EGFR, SRC, and HSP90AA1. RT-qPCR and Western blot analyses further confirmed that PA treatment significantly decreased the mRNA and protein expression of AKT1, EGFR, and SRC while increasing the HSP90AA1 mRNA and protein levels. ConclusionIt was suggested that PA may exert its anti-NB effects by inhibiting AKT1, EGFR, and SRC expression, potentially modulating the PI3K/AKT signaling pathway. These findings provide crucial evidence supporting PA’s development as a therapeutic candidate for NB.
6.Development and validation of a multidisciplinary risk assessment scale for immune checkpoint inhibitor-associated myocarditis
Yanan DAI ; Yuan LIU ; Yuchen XU ; Qingqing CAI ; Yan WANG ; Yuhong ZHOU ; Leilei CHENG ; Junbo GE
Chinese Journal of Clinical Medicine 2025;32(4):561-567
Objective To develop a risk assessment scale for immune checkpoint inhibitor (ICI)-associated myocarditis based on multidisciplinary collaboration, and to evaluate its diagnostic performance. Methods Based on multidisciplinary cooperation, integrating clinical experience from oncology and cardiology, literature data, and patient conditions, a risk assessment scale for ICI-associated myocarditis was developed. A total of 101 patients with malignancies who received immunotherapy at Zhongshan Hospital, Fudan University, from October 2020 to October 2024 were included as the validation cohort. Patients were stratified into low-risk (0-1 point), medium-risk (2-4 points), and high-risk (≥5 points) groups based on their scale scores. The association between pretictive risk stratifications and actual assessment results was assessed using the Cox proportional hazards regression model. The predictive value of the scale for ICI-associated myocarditis was evaluated using receiver operating characteristic (ROC) curve. Agreement between the scale scores and actual assessment results was assessed using Cohen’s Kappa coefficient. Results Based on the scale pretictive results, 28(27.7%), 8(7.9%), 65(64.4%) patients were at low risk, medium risk, and high risk for ICI-related myocarditis, respectively; however, 46(45.5%), 8(7.9%), 47(46.5%) were at low risk, medium risk, and high risk actually. Kaplan-Meier survival analysis showed that the cumulative incidence of ICI-related myocarditis in the high-risk group was significantly higher than that in the medium- and low-risk groups (P<0.05). In the multivariable-adjusted Cox proportional hazards model, the ICI-related myocarditis risk in high-risk group was about 4 times that in the low-risk group. ROC curve analysis demonstrated that the average area under the curve (AUC) for predicting ICI-related myocarditis was 0.81, with an accuracy of 0.74. The Cohen’s Kappa coefficient was 0.55, indicating moderate agreement. In the actual high-risk group, no patient was predicted to be at low risk; in the actual low-risk group, 16 patients were predicted to be at high risk. Conclusions This risk assessment scale for ICI-associated myocarditis shows high predictive performance. It provides oncologists with a simple yet effective multidisciplinary diagnostic reference tool, potentially enhancing early identification of ICI-associated myocarditis.
7.Mechanism Prediction of Banxia Baizhu Tianmatang and Danggui Shaoyaosan Intervention in Ménière's Disease Based on LC-MS Technology Combined with Network Bioinformatics
Xingye ZHU ; Jiaxiang YU ; Ziyue YUAN ; Shengrong GUO ; Jianyu DAI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(24):50-60
ObjectiveThis study aims to analyze the pharmacodynamic material basis and multi-target mechanism of action of Banxia Baizhu Tianmatang combined with Danggui Shaoyaosan in the treatment of Meniere's disease(MD). MethodsUltra-performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) (mobile phase: gradient elution with 0.1% formic acid aqueous solution-acetonitrile. Mass spectrometry scanning range: m/z 90-1 300) was used to identify the chemical components of the compound recipe and components absorbed into blood. The core mechanism was predicted by combining network pharmacology (target screening via SwissTargetPrediction and GeneCards databases, and construction of protein-protein interaction (PPI) network by STRING) and molecular docking (evaluated by Autodock, with binding energy ≤ -5.0 kcal·mol-1). For animal experiment validation, 36 Sprague Dawley (SD) rats were divided into a blank group, a model group (postauricular injection of lipopolysaccharide (LPS) at 1 mg·kg-1), low/medium/high-dose Chinese medicine groups (5.94, 11.88, and 23.76 g·kg-1·d-1, respectively), and Western medicine group (betahistine at 0.1 mg·kg-1·d-1). After eight weeks of intervention, the gene and protein expressions in cochlear tissue were detected. Results①A total of 2 831 chemical components and 173 components absorbed into blood were identified, with terpenoids showing the highest absorption rate into blood(10.28%). ②60 common drug-disease targets were screened, with core targets including tumor necrosis factor-α(TNF-α),interleukin-6(IL-6), Toll-like receptor 4(TLR4), angiotensin-converting enzyme(ACE), and endothelial nitric oxide synthase 3(NOS3).These targets were enriched in the nuclear factor-κB(NF-κB) signaling pathway and renin-angiotensin system(P<0.05). Molecular docking showed that the active component YC-1 had a strong binding ability to TNF(binding energy-9.66 kcal·mol-1). ③In animal experiments, the high-dose Chinese medicine group significantly down-regulated the expression of pro-inflammatory factors TNF mRNA(P<0.01)and up-regulated vascular regulatory factors NOS3 protein(P<0.01), and alleviated cochlear pathological damage[hematoxylin eosin (HE) score: from 4 to 2]. ConclusionThis compound recipe synergistically regulates the TNF/NF-κB inflammatory pathway and ACE/NOS3 vascular homeostasis pathway through flavonoids, triterpenoids, and other components, thereby inhibiting endolymphatic hydrops and cochlear damage. It provides a scientific basis for the theory of "simultaneous treatment of phlegm and blood stasis" in traditional Chinese medicine.
8.Efficacy of Tiaogan Huaxian Pills in Prevention and Treatment of Hepatic Fibrosis in Hepatitis B and Its Effect on Diffusion-weighted Imaging
Haiyang DAI ; Guye HUANG ; Wencong XU ; Yan SHEN ; Yuan LI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):159-165
ObjectiveTo investigate the preventive and therapeutic effects of Tiaogan Huaxian pills combined with entecavir on hepatic fibrosis in chronic hepatitis B with liver Qi stagnation, spleen deficiency, and blood stasis syndrome and its effect on diffusion-weighted imaging (DWI). MethodClinical data of 117 patients with liver disease who visited the Department of Hepatology at the First Affiliated Hospital of Guangxi University of Chinese Medicine from January 2021 to April 2022 were retrospectively analyzed. According to different treatment plans, they were divided into a control group (59 cases) and a treatment group (58 cases). Both groups of patients received entecavir-based etiology treatment, and the treatment group added Tiaogan Huaxian pills on the basis of basic treatment. Both groups were treated for 24 weeks. Before and after treatment, the two groups were compared in terms of alanine aminotransferase (ALT), advanced surgical technologies (AST), total bilirubin (TBil), hepatitis B virus (HBV)-DNA conversion rate, liver stiffness measurement (LSM), four items of liver fibrosis (hyaluronidase, type Ⅲ pro-collagen, type Ⅳ collagen, and laminin), the fibrosis index based on four factors (FIB-4), the aspartate aminotransferase to platelet ratio index (APRI), the apparent diffusion coefficient (ADC) value in magnetic resonance imaging (MRI), and traditional Chinese medicine symptom scores, so as to analyze the efficacy of the two groups. ResultBefore treatment, there was no significant difference in ALT, AST, TBil, LSM, four items of liver fibrosis, FIB-4, APRI, HBV-DNA conversion rate, ADC value, and traditional Chinese medicine symptom scores between the two groups. After treatment, both groups of patients showed significant reductions in ALT, AST, TBil, LSM, hyaluronidase, type Ⅲ pro-collagen, type Ⅳ collagen, laminin, FIB-4, and APRI (P<0.05) and a significant increase in ADC value (P<0.05) and HBV-DNA conversion rate (P<0.01). The traditional Chinese medicine symptom score of the treatment group decreased significantly (P<0.05). Compared with the control group after treatment, the effective rate of clinical traditional Chinese medicine in the treatment group was 91.38% (53/58), which was significantly higher than that of the control group (54.23%, 32/59) (Z=-4.325, P<0.01). In the treatment group, ALT, AST, TBil, LSM, hyaluronidase, type Ⅲ pro-collagen, type Ⅳ collagen, laminin, FIB-4, APRI, and traditional Chinese medicine symptom scores all decreased significantly (P<0.05), and the increase in ADC values was more significant (P<0.05), while the difference in HBV-DNA conversion rate was not statistically significant. There were no serious adverse reactions or events in either group. ConclusionTiaogan Huaxian pills combined with entecavir have significant clinical efficacy in the treatment of hepatic fibrosis in chronic hepatitis B, which can reduce liver inflammation activity, delay hepatic fibrosis progression, and reduce traditional Chinese medicine symptom scores. It is worthy of clinical promotion and application.
9.Optical coherence tomography angiography manifestations of different degrees of nonproliferative diabetic retinopathy
Menghuan DAI ; Dongli LI ; Yixin CUI ; Lu SHEN ; Qirui LI ; Lingna RAO ; Shurong GUO ; Ling YUAN
International Eye Science 2024;24(2):307-311
AIM: To investigate the differences in varying stages of non-proliferative diabetic retinopathy(NPDR)using optical coherence tomography angiography(OCTA).METHODS: Cross-sectional study. A total of 77 cases(77 eyes)of diabetic patients were included, and they were divided into non-diabetic retinopathy(NDR; 23 eyes)and non-proliferative diabetic retinopathy(NPDR; 54 eyes)groups, further subdivided into mild NPDR(20 eyes), moderate NPDR(20 eyes), and severe NPDR(14 eyes). Foveal avascular zone(FAZ)area, superficial and deep capillary plexus densities(SSP and DSP), and visual acuity(LogMAR)were compared between NDR and NPDR groups. Furthermore, the visual acuity, FAZ area and levels of SSP and DSP were compared in different degrees of NPDR. Correlation analysis were conducted to elucidate relationships between FAZ area, visual acuity, SSP, DSP, and severity of the disease.RESULTS: Compared with the NDR group, the visual acuity(LogMAR)and macular FAZ area increased, while SSP and DSP were decreased in the NPDR group(P<0.05); there were significant differences in visual acuity, FAZ area and SSP and DSP levels in different degrees of NPDR(P<0.05). Visual acuity(LogMAR)and FAZ area displayed a positive correlation with the severity of disease, while SSP and DSP showed a negative correlation.CONCLUSION: With the progression of NPDR, the visual acuity(LogMAR)and FAZ area increased, and the SSP and DSP decreased.
10.Experimental study of Lentinan antagonizes against hepatic lipid deposition induced by sodium arsenite in mice
Zongying DAI ; Qianqian WU ; Yuan YANG
Journal of Public Health and Preventive Medicine 2024;35(1):20-23
Objective To investigate the ameliorative effect of Lentinan (LNT) on sodium arsenite (SA)-induced hepatic lipid deposition in mice. Methods C57BL/6 mice were used as the experimental subjects, which were divided into control group, SA-exposed group, LNT + SA-exposed group and LNT control group. Blood and liver tissue samples were collected at the end of the experiment, and serum glutathione transaminase (ALT) and glutathione aminotransferase (AST) levels were detected by enzyme-linked immunosorbent assay (ELISA). A part of liver tissues was stained with hematoxylin-eosin (HE) or oil red O to observe the characteristics of liver pathological damage and lipid deposition, and another part of liver tissues was used to detect triglyceride (TG) and Adiponectin (APN) levels by ELISA. Results Compared with control group or LNT control group, SA-exposed group showed the increased levels of AST and ALT, showing the characteristics of liver histopathological damage and lipid deposition, and the APN level decreased while the TG level increased (P<0.05). Compared with SA-exposed group, the levels of AST and ALT decreased in LNT + SA-exposed group, showing the reduced degree of liver tissue damage and lipid deposition, and APN level upregulated while TG level downregulated (P<0.05). Conclusion Chronic SA exposure induces liver function damage, APN downregulation and lipid deposition in C57BL/6 mice, while LNT intervention leads to the significantly improvement of hepatic damage and lipid deposition, which may be related to the elevated APN level in liver.


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