1.Application of Zebrafish in Quality Evaluation of Traditional Chinese Medicine
Jiaqi LI ; Xiaolu CHEN ; Jiarui WU ; Xiaoyu TAO ; Qiqi FAN ; Shengyun DAI ; Chongjun ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):58-70
As a new type of model organism, zebrafish is gradually gaining prominence in the field of scientific research. The unique biological characteristics and advantages of zebrafish make them play an increasingly important role in the quality evaluation of traditional Chinese medicine. Compared with other common experimental animals, zebrafish have a fast reproductive and growth speed and high embryo transparency, making them an ideal model for evaluating the quality of traditional Chinese medicine. This provides a new perspective and method for research on traditional Chinese medicine. With the growing global interest in traditional Chinese medicine, it has become crucial to find scientific and accurate methods to evaluate the quality and effectiveness of traditional Chinese medicine. The introduction of the zebrafish model has brought new breakthroughs in the quality evaluation of traditional Chinese medicine. To further promote the application of zebrafish in evaluating the quality of traditional Chinese medicine, this article systematically searched and sorted out the previous studies related to the application of zebrafish for this purpose since 2023. The commonly used disease models and indicators of zebrafish in evaluating the effectiveness of traditional Chinese medicine, as well as the mechanism of zebrafish in exploring the active ingredients of traditional Chinese medicine, were primarily reviewed. The application of zebrafish in evaluating the safety of traditional Chinese medicine and the typical examples in ensuring the quality of traditional Chinese medicine were demonstrated. The limitations encountered by zebrafish models in evaluating the quality of traditional Chinese medicine were highlighted. The resolution of these problems will help further improve the accuracy and reliability of zebrafish in evaluating the quality of traditional Chinese medicine. The article discussed the evaluation of effectiveness, safety, and quality control of zebrafish applied in traditional Chinese medicine, so as to provide a reference for establishing standards for traditional Chinese medicine and promoting its modernization in the future.
2.Evaluation of Safety of Compound Kushen Injection and Efficacy in Treatment of Inflammatory Bowel Disease Based on Zebrafish Model
Xiaolu CHEN ; Jiaqi LI ; Linzhen CHEN ; Qi CHEN ; Zhiqiang MA ; Chongjun ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):71-78
ObjectiveTo evaluate the safety/efficacy of compound Kushen injection (CKI) by zebrafish model and explore the possible mechanism. MethodsZebrafish were exposed to different concentrations of CKI solution, and the mortality rate after 24 h was calculated. After exposure to sublethal concentration (
3.Hepatotoxicity and Anti-osteoporotic Mechanism of Evodiamine
Jiaqi LI ; Xiaolu CHEN ; Xiaoyu TAO ; Shan LU ; Qiqi FAN ; Jiarui WU ; Chongjun ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):79-86
ObjectiveBased on the zebrafish model, the hepatotoxicity and anti-osteoporotic activity of evodiamine (EVO) were studied. The mechanism of EVO in treating osteoporosis was explored by using network pharmacology and real-time polymerase chain reaction(Real-time PCR). MethodsThree days after fertilization (3 dpf), zebrafish were randomly selected and exposed to different concentrations of EVO solution for 96 hours. The mortality rate of zebrafish at different concentrations was calculated at the exposure endpoint, and a "dose-toxicity" curve was drawn. The 10% lethal concentration (LC10) was calculated. Liver phenotype, acridine orange staining, and pathological tissue sections of liver-transgenic zebrafish [CZ16 (gz15Tg.Tg (fabp 10a: ds Red; ela31: EGFP))] were used to confirm hepatotoxicity of EVO. On this basis, prednisolone was used to create a model of osteoporosis in zebrafish. The skull development, area of the skull stained by alizarin red, and cumulative optical density were used as indicators to evaluate the anti-osteoporotic activity of EVO in a safe dose. Based on network pharmacology, the mechanism of action of EVO in the treatment of osteoporosis was predicted and verified through Real-time PCR. ResultsThe LC10 of EVO on zebrafish (7 dpf) was determined to be 0.4 mg·L-1. Compared with the control group, sublethal concentrations (
4.The interaction between polyphyllin I and SQLE protein induces hepatotoxicity through SREBP-2/HMGCR/SQLE/LSS pathway
Zhiqi LI ; Qiqi FAN ; Meilin CHEN ; Ying DONG ; Farong LI ; Mingshuang WANG ; Yulin GU ; Simin GUO ; Xianwen YE ; Jiarui WU ; Shengyun DAI ; Ruichao LIN ; Chongjun ZHAO
Journal of Pharmaceutical Analysis 2023;13(1):39-54
Polyphyllin Ⅰ(PPⅠ)and polyphyllin Ⅱ(PⅡ)are the main active substances in the Paris polyphylla.However,liver toxicity of these compounds has impeded their clinical application and the potential hepatotoxicity mechanisms remain to be elucidated.In this work,we found that PPⅠ and PⅡ exposure could induce significant hepatotoxicity in human liver cell line L-02 and zebrafish in a dose-dependent manner.The results of the proteomic analysis in L-02 cells and transcriptome in zebrafish indicated that the hepa-totoxicity of PPⅡ and PⅡwas associated with the cholesterol biosynthetic pathway disorders,which were alleviated by the cholesterol biosynthesis inhibitor lovastatin.Additionally,3-hydroxy-3-methy-lglutaryl CoA reductase(HMGCR)and squalene epoxidase(SQLE),the two rate-limiting enzymes in the choles-terol synthesis,selected as the potential targets,were confirmed by the molecular docking,the over-expression,and knockdown of HMGCR or SQLE with siRNA.Finally,the pull-down and surface plasmon resonance technology revealed that PPⅠ could directly bind with SQLE but not with HMGCR.Collectively,these data demonstrated that PPⅠ-induced hepatotoxicity resulted from the direct binding with SQLE protein and impaired the sterol-regulatory element binding protein 2/HMGCR/SQLE/lanosterol synthase pathways,thus disturbing the cholesterol biosynthesis pathway.The findings of this research can contribute to a better understanding of the key role of SQLE as a potential target in drug-induced hepatotoxicity and provide a therapeutic strategy for the prevention of drug toxic effects with similar structures in the future.
5.Study on Mechanism of Action of Matrine in Treatment of Inflammatory Bowel Disease Based on Zebrafish Model and Network Pharmacology
Linzhen CHEN ; Xue ZHANG ; Qi CHEN ; Xue YU ; Shengyun DAI ; Zhiqiang MA ; Chongjun ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(22):88-94
ObjectiveTo study the mechanism of matrine in the treatment of inflammatory bowel disease (IBD) based on the zebrafish model and network pharmacology. MethodThe IBD model of zebrafish was established using 2,4,6-trinitro-benzenesulfonicacid (TNBS), and the intestinal phagocytic function, goblet cell secretion, and neutrophil aggregation were evaluated using neutral red staining, alcian blue staining, and neutrophil number changes. Changes in tumor necrosis factor (TNF)-α and cholecystokinin (CCK) content in zebrafish were determined by using relevant reagent kits. Network pharmacology and molecular docking techniques were used to predict the potential mechanism of matrine in the treatment of IBD. Gene expression of relevant targets was verified through Real-time polymerase chain reaction (Real-time PCR). ResultCompared with the model group, the matrine administration group can increase the neutral red staining area in a dose-dependent manner and improve intestinal phagocytic function(P<0.05,P<0.01). It can reduce the staining area of alcian blue and affect the secretion of intestinal goblet cells(P<0.01). It can reduce the number of neutrophil granulocytes, relieve its aggregation, significantly reduce TNF-α content(P<0.01), and increase the CCK content. Network pharmacology analysis identifies 28 potential targets for matrine in the treatment of IBD. The top five targets by protein-protein interaction (PPI) network analysis are CHRNA7, DRD1, CHRNA4, SLC6A3, and GRM5. The Kyoto encyclopedia of genes and genomes (KEGG) results show that the treatment of IBD with matrine may be related to neuroactive ligand-receptor interaction, cholinergic synapse, and neutrophil extracellular trap formation. Real-time PCR results show that matrine can affect the expression level of related target genes. Conclusionmatrine has a certain therapeutic effect on IBD and can affect the inflammatory response of IBD. Its therapeutic effect may be related to neuroactive ligand-receptor interaction and other pathways.
6.Influence of prior percutaneous coronary intervention on outcome of coronary artery bypass grafting: A multi-center clinical study
Hongwei JIANG ; Hang ZHANG ; Wen CHEN ; Fangjing ZHENG ; Yongfeng SHAO ; Yongxiang QIAN ; Guoxiang WANG ; Mingqiu LI ; Qingsheng YOU ; Zhiyong LIU ; Yong WANG ; Zhenya SHEN ; Wei LI ; Demin LI ; Su HUANG ; Chongjun ZHONG ; Rui WANG ; Xin CHEN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(11):1436-1441
Objective To investigate the influence of prior percutaneous coronary intervention (PCI) on the outcome of coronary artery bypass grafting (CABG). Methods Clinical data of 5 216 patients from Jiangsu Province CABG registry who underwent primary isolated CABG from 2016 to 2019 were retrospectively analyzed. Patients were divided into a PCI group (n=673) and a non-PCI group (n=4 543) according to whether they had received PCI treatment. The PCI group included 491 males and 182 females, aged 62.6±8.2 years, and the non-PCI group included 3 335 males and 1 208 females, aged 63.7±8.7 years. Multivariable logistic regression and propensity score matching (PSM) were used to compare 30-day mortality, incidence of major complications and 1-year follow-up outcomes between the two groups. Results Both in original cohort and matched cohort, there was no statistical difference in the 30-day mortality [14 (2.1%) vs. 77 (1.7%), P=0.579; 14 (2.1%) vs. 11 (1.6%), P=0.686], or the incidence of major complications (myocardial infarction, stroke, mechanical ventilation≥24 h, dialysis for new-onset renal failure, deep sternal wound infection and atrial fibrillation) (all P>0.05). The rate of reoperation for bleeding in the PCI group was higher than that in the non-PCI group [19 (2.8%) vs. 67 (1.5%), P=0.016; 19 (2.8%) vs. 7 (1.0%), P=0.029]. Both in original cohort and matched cohort, there was no statistical difference in 1-year survival rate between the two groups [613 (93.1%) vs. 4 225 (94.6%), P=0.119; 613 (93.1%) vs. 630 (95.2%), P=0.124], while the re-admission rate in the PCI group was significantly higher than that in the non-PCI group [32 (4.9%) vs. 113 (2.5%), P=0.001; 32 (4.9%) vs. 17 (2.6%), P=0.040]. Conclusion This study shows that a history of PCI treatment does not significantly increase the perioperative mortality and major complications of CABG, but increases the rate of cardiogenic re-admission 1 year postoperatively.
7.Clinical effects of high resolution magnetic resonance imaging examination on the evaluation for lower rectal cancer undergoing sphincter-preserving operation
Bangfei CHEN ; Chaoying YANG ; Chun JIN ; Chongjun ZHOU
Chinese Journal of General Surgery 2020;35(8):616-619
Objective:By comparing preoperative high resolution magnetic resonance imaging (MRI) examination with postoperative pathologic results, to investigate the effects of MRI examination evaluation on the anatomical level and clinical outcome.Methods:We conducted a retrospective study on 72 patients who underwent resection of rectal cancer at the Second Affiliated Hospital of Wenzhou Medical University between Apr 2017 and Nov 2018, including 35 patients undergoing laparoscopic resection and 37 patients doing open resection. All cases received high resolution MRI examination before operation. The diagnostic accuracy of MRI, operation safety, and the short-term outcomes were analyzed.Results:There were no postoperative tumor recurrence. The accuracy rate of preoperative MRI evaluation of T stage was 85%, and positive N+ was 74%. There were no difference in postoperative complications between the open resection group and laparoscopic resection group (29% vs. 22%, χ 2=0.463, P=0.496). The proximal and distal margin was negative, postoperative circumferential resection margin and preoperative mesorectal fascia was consistent, the distance between the lower margin of the tumor and the anal right angle measured by MRI were consistent with the distance between the tumor from the dentate line. Conclusion:High resolution MRI with a good tissue resolution, has a high preoperative diagnosis accuracy for T and N staging of the low rectal cancer, with decisive role in the evaluation on the anatomical level, improving the quality and safy of surgery.
8.Content Determination and Difference Analysis of 3 Saponins in Different Parts of Ypsilandra thibetica from Different Producing Areas
Shuai CHEN ; Mei ZHANG ; Chongjun YUAN ; Sen LUO ; Jia WANG
China Pharmacy 2020;31(3):325-329
OBJECTIVE:To establish the method for content dete rmination of polyphyllin Ⅱ,Ⅵ,Ⅶ in Ypsilandra thibetica , and to compare the differences of 3 saponins in different parts of Y. thibetica from different producing areas. METHODS :HPLC method was adopted to determine the contents of polyphyllin Ⅱ,Ⅵ,Ⅶ in whole grass part and underground part of Y. thibetica from 10 producing areas. The determination was performed on Kromasil C 18column with mobile phase consisted of acetonirile-water (gradient elution )at the flow rate of 1.0 mL/min. The column temperature was 35 ℃,and detection wavelength was set at 203 nm;sample size was 10 μL. With the contents of 3 saponins as the index ,20 batches of Y. thibetica were analyzed by cluster analysis and PLS-DA analysis ;the aggregation of samples was analyzed and determined the primary difference components. RESULTS:The linear range of polyphyllin Ⅱ,polyphyllin Ⅵ and polyphyllin Ⅶ were 0.051-2.04,0.007-0.28,0.168-6.72 μg, respectively(r≥0.999 5);the detection limits were 1.92,1.75,1.87 ng,respectively;and the quantitative limits were 6.40,5.87, 6.23 ng,respectively;RSD of precision ,reproducibility and stability tests (24 h)were all lower than 2%(n=6);the average recovery rates were 99.29%,101.38% and 99.64%,with RSDs of 1.17%,2.64%,0.75%(n=6),respectively. The content of polyphyllin Ⅱ was 0.615-1.875 mg/g,that of polyphyllin Ⅵ was 0-0.095 mg/g,and that of polyphyllin Ⅶ was 3.158-12.354 mg/g. Cluster analysis showed that 20 batches of samples were clustered into two groups ,batch S 9-S12 were clustered in to one group,and the other 16 batches of samples were clustered into another group. PLS-DA analysis showed that 20 batches of samples were divided into 3 areas,batch S 1,S2,S8,S14,S16,S20 were included in area Ⅰ;batch S 9-S12 included in area Ⅱ;and the rest of the samples included in area Ⅲ. The quality of Y. thibetica from different habitats was different ,and there was no difference in the saponin composition between the whole grass and the underground part. Weight ranking found that mail:cscdtcm@126.com polyphyllin Ⅶ was the main difference component in Y. thibetica,and the content of polyphyllin Ⅶ in Y. thibetica from Pengzhou city and Dayi county was the highest. CONCLUSIONS :The established method is simple ,convenient and sensitive. It can be used for the content determination of 3 saponins in Y. thibetica . The content of active components is higher and the quality is better in Y. thibetica from Pengzhou city and Dayi county.
9.Jiangsu Province Coronary Artery Bypass Grafting Registry study: a report of 4 661 patients in 13 centers
Rui WANG ; Yongfeng SHAO ; Yongxiang QIAN ; Guoxiang WANG ; Mingqiu LI ; Qingsheng YOU ; Zhiyong LIU ; Yong WANG ; Zhenya SHEN ; Wei LI ; Demin LI ; Su HUANG ; Chongjun ZHONG ; Xin CHEN
Chinese Journal of Surgery 2020;58(5):350-355
Objective:To examine the overall status of the Jiangsu Province Coronary Artery Bypass Grafting Registry database.Methods:The patients date of Jiangsu Province Coronary Artery Bypass Grafting Registry database from October 2017 to December 2019 was collected retrospectively.Risk factors, history, cardiac function (New York Heart Association class), extent of coronary artery lesion, European system for cardiac operative risk evaluation Ⅱ (EuroSCORE Ⅱ), cardiopulmonary bypss, arterial grafts, the numbers and flow of grafts and postoperative major adverse cardiac and cerebrovascular event(MACCE) information were analyzed. The clinical data of patients underwent on-pump CABG(ONCABG) or off-pump CABG (OPCAB) were compared by t test or χ 2 test. Results:Up till December 2019, the database enrolled 7 138 patients, in which 4 661 patients receiving primary isolated CABG. There were 3 486 males and 1 175 females with the age of (64.6±8.1) years (range:31 to 87 years). There were coronary left main disease in 960 patients, triple vessel disease in 3 934 patients, both left main and triple vessel disease in 837 patients, ejection fraction >50% in 3 841 patients, cardiac function class Ⅲ to Ⅳ in 1 664 patients. EuroSCORE Ⅱ was (2.3±0.7)% (range: 0.5% to 35.8%). There were 2 731 patients (58.59%) underwent ONCABG and 1 930 patients (41.41%) underwent OPCAB. There were 4 144 patients (88.91%) for whom the left internal thoracic artery was harvested. Seven centers (2 centers routinely) used left radial artery, 5 centers (3 centers routinely) used the transit time flow meter. The graft was 3.4±0.7 (range:1 to 7), the aortic crossclamp time was (65.0±20.4) minutes (range: 21 to 196 minutes), the cardiopulmonary bypass time was (90.0±24.2) minutes (range: 33 to 227 minutes). In-hospital death ocurred in 84 patients(1.80%), while re-operation in 93 patients (2.00%), myocardial infarction in 71 patients (1.52%), cerebral infarction in 33 patients (0.71%) and dialysis in 56 patients (1.20%). There were 2 936 patients prescribed with secondary prevention drugs(62.99%).Comparing with OPCAB group, ONCABG group had younger age, more female, more diabetes mellitus, more history of myocardial infarction and percutaneous transluminal coronary angioplasty, poorer cardiac function and coronary lesions, higher EuroSCORE Ⅱ, preoperatively (all P<0.05), and was associated with higher MACCE (135/2 731 vs. 71/1 930, χ 2=4.280, P=0.039), and of more grafts, transfusion and intra-aortic balloon counterpulsation application (all P<0.05). Conclusions:Jiangsu Province Coronary Artery Bypass Grafting Registry database is generally in good operation, and some parameters still need to be improved. Comparing with OPCAB group, ONCABG has more severe preoperative general conditions, while the outcomes is acceptable.
10.Jiangsu Province Coronary Artery Bypass Grafting Registry study: a report of 4 661 patients in 13 centers
Rui WANG ; Yongfeng SHAO ; Yongxiang QIAN ; Guoxiang WANG ; Mingqiu LI ; Qingsheng YOU ; Zhiyong LIU ; Yong WANG ; Zhenya SHEN ; Wei LI ; Demin LI ; Su HUANG ; Chongjun ZHONG ; Xin CHEN
Chinese Journal of Surgery 2020;58(5):350-355
Objective:To examine the overall status of the Jiangsu Province Coronary Artery Bypass Grafting Registry database.Methods:The patients date of Jiangsu Province Coronary Artery Bypass Grafting Registry database from October 2017 to December 2019 was collected retrospectively.Risk factors, history, cardiac function (New York Heart Association class), extent of coronary artery lesion, European system for cardiac operative risk evaluation Ⅱ (EuroSCORE Ⅱ), cardiopulmonary bypss, arterial grafts, the numbers and flow of grafts and postoperative major adverse cardiac and cerebrovascular event(MACCE) information were analyzed. The clinical data of patients underwent on-pump CABG(ONCABG) or off-pump CABG (OPCAB) were compared by t test or χ 2 test. Results:Up till December 2019, the database enrolled 7 138 patients, in which 4 661 patients receiving primary isolated CABG. There were 3 486 males and 1 175 females with the age of (64.6±8.1) years (range:31 to 87 years). There were coronary left main disease in 960 patients, triple vessel disease in 3 934 patients, both left main and triple vessel disease in 837 patients, ejection fraction >50% in 3 841 patients, cardiac function class Ⅲ to Ⅳ in 1 664 patients. EuroSCORE Ⅱ was (2.3±0.7)% (range: 0.5% to 35.8%). There were 2 731 patients (58.59%) underwent ONCABG and 1 930 patients (41.41%) underwent OPCAB. There were 4 144 patients (88.91%) for whom the left internal thoracic artery was harvested. Seven centers (2 centers routinely) used left radial artery, 5 centers (3 centers routinely) used the transit time flow meter. The graft was 3.4±0.7 (range:1 to 7), the aortic crossclamp time was (65.0±20.4) minutes (range: 21 to 196 minutes), the cardiopulmonary bypass time was (90.0±24.2) minutes (range: 33 to 227 minutes). In-hospital death ocurred in 84 patients(1.80%), while re-operation in 93 patients (2.00%), myocardial infarction in 71 patients (1.52%), cerebral infarction in 33 patients (0.71%) and dialysis in 56 patients (1.20%). There were 2 936 patients prescribed with secondary prevention drugs(62.99%).Comparing with OPCAB group, ONCABG group had younger age, more female, more diabetes mellitus, more history of myocardial infarction and percutaneous transluminal coronary angioplasty, poorer cardiac function and coronary lesions, higher EuroSCORE Ⅱ, preoperatively (all P<0.05), and was associated with higher MACCE (135/2 731 vs. 71/1 930, χ 2=4.280, P=0.039), and of more grafts, transfusion and intra-aortic balloon counterpulsation application (all P<0.05). Conclusions:Jiangsu Province Coronary Artery Bypass Grafting Registry database is generally in good operation, and some parameters still need to be improved. Comparing with OPCAB group, ONCABG has more severe preoperative general conditions, while the outcomes is acceptable.

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