1.Application of the three-guidewire technique in retrograde stent placement for mucosal sleeve avulsion of the lower ureter
Hongchao DONG ; Yu HAO ; Zhao NI ; Qinzhang WANG
Journal of Modern Urology 2026;31(2):163-167
Objective To evaluate the feasibility and efficacy of the three-guidewire technique for retrograde stent placement in the treatment of ureteral mucosal sleeve avulsion (UMSA). Methods A retrospective analysis was conducted on the clinical data of 12 patients with UMSA secondary to ureteroscopic lithotripsy for ureteral calculi in our hospital during Jul. 2010 and Oct. 2023. The three-guidewire technique was employed for retrograde stent placement during the procedure. A secondary ureteroscopic lithotripsy was performed one month postoperatively.Results Double-J stent was successfully placed in all 12 patients, with a mean stent insertion time of 23.3(16-30)minutes. No intraoperative exacerbation of mucosal avulsion occurred. One patient was lost to follow-up. At the one-month postoperative ureteroscopic examination and lithotripsy, 81.8% (9/11) cases exhibited unobstructed ureters, while 18.2%(2/11) developed annular membranous strictures, which were resolved following dilation. During subsequent follow-up, the postoperative creatinine level after double-J stent removal showed a mild decrease compared to the preoperative level [87.9(72-100)μmol/L vs. 90.1(69-108)μmol/L]. Up to 90.9%(10/11) patients remained free of hydronephrosis, with only 1 case (9.1%) requiring laparoscopic ureterovesical reimplantation for hydronephrosis 4 months postoperatively. Conclusion The three-guidewire technique for retrograde stent placement in the treatment of UMSA demonstrates strong operability, high success rates, favorable outcomes, and a low incidence of ureteral stricture, which warrant wide clinical promotion.
2.Quantitative analysis and distribution comparison of salidroside and its liposomes in mouse serum,bronchoalveolar lavage fluid,and lung tissue
Rong WANG ; Ni ZHAO ; Guoquan LI ; Yafeng WANG ; Shen’ao LI ; Jiaming LAI ; Hao CAI
China Pharmacy 2026;37(15):1998-2002
OBJECTIVE Establish a quantitative analysis method for salidroside (Sal) in mouse serum, bronchoalveolar lavage fluid (BALF), and lung tissue, and compare the distribution differences of Sal and its liposomes. METHODS Using high-performance liquid chromatography(HPLC), Welch Xtimate C18 was used as the chromatographic column, with methanol-formic acid aqueous solution (formic acid volume fraction: 0.1% for serum and BALF detection, 0.2% for lung tissue detection) as the mobile phase, flow rate of 1.0 mL/min, detection wavelength of 275 nm, column temperature of 30 ℃, and injection volume of 10 μL. Serum, BALF, and lung tissue homogenate samples were precipitated with methanol for protein analysis. Twelve mice were randomly divided into Sal group and Sal liposome group, and the concentrations of Sal in serum, BALF, and lung tissue were detected after intragastric administration corresponding liquid medicine at a dose of 300 mg/kg (calculated as Sal) for 0.5 h. RESULTS The linear ranges of Sal in serum, BALF, and lung tissue were 1.56-200.00 μg/mL, 1.56-100.00 μg/mL, and 6.24-400.00 μg/g, respectively (all r=0.999 9), with intra-day RSD of 1.35%-3.87%, 1.10%-2.75%, and 1.90%-3.86%, respectively; the inter-day RSD of 3.38%-5.11%, 2.22%- 4.36%, and 2.41%-4.73%, respectively. The relative accuracies were 97.95%-101.61%, 98.29%-101.82%, and 97.41%- 106.94%, respectively. The stability RSDs were all less than 6%. Compared with the Sal group, the Sal liposome group significantly increased the concentration of Sal in BALF of mice (P<0.05), and significantly decreased the concentration of Sal in serum and lung tissue (P<0.05). CONCLUSIONS The HPLC method established in this study is suitable for quantitative analysis of Sal in mouse serum, BALF, and lung tissue. Sal liposomes have a tendency to distribute towards the alveolar cavity during early administration.
3.Updates and amendments of the Chinese Pharmacopoeia 2025 Edition (Volume Ⅰ)
LI Hao ; SHEN Mingrui ; ZHANG Pang ; ZHAI Weimin ; NI Long ; HAO Bo ; ZHAO Yuxin ; HE Yi ; MA Shuangcheng ; SHU Rong
Drug Standards of China 2025;26(1):017-022
The Chinese Pharmacopoeia is the legal technical standard which should be followed during the research, production, use, and administration of drugs. At present, the new edition of the Chinese Pharmacopoeia is planned to be promulgated and implemented. This article summarizes and analyzes the main characteristics and the content of updates and amendments of the Chinese Pharmacopoeia 2025 Edition(Volume Ⅰ), to provide a reference for the correct understanding and accurate implementation the new edition of the pharmacopoeia.
4.Xinyang Tablets ameliorate ventricular remodeling in heart failure via FTO/m6A signaling pathway.
Dong-Hua LIU ; Zi-Ru LI ; Si-Jing LI ; Xing-Ling HE ; Xiao-Jiao ZHANG ; Shi-Hao NI ; Wen-Jie LONG ; Hui-Li LIAO ; Zhong-Qi YANG ; Xiao-Ming DONG
China Journal of Chinese Materia Medica 2025;50(4):1075-1086
The study was conducted to investigate the mechanism of Xinyang Tablets( XYP) in modulating the fat mass and obesity-associated protein(FTO)/N6-methyladenosine(m6A) signaling pathway to ameliorate ventricular remodeling in heart failure(HF). A mouse model of HF was established by transverse aortic constriction(TAC). Mice were randomized into sham, model, XYP(low, medium, and high doses), and positive control( perindopril) groups(n= 10). From day 3 post-surgery, mice were administrated with corresponding drugs by gavage for 6 consecutive weeks. Following the treatment, echocardiography was employed to evaluate the cardiac function, and RT-qPCR was employed to determine the relative m RNA levels of key markers, including atrial natriuretic peptide( ANP), B-type natriuretic peptide( BNP), β-myosin heavy chain(β-MHC), collagen type I alpha chain(Col1α), collagen type Ⅲ alpha chain(Col3α), alpha smooth muscle actin(α-SMA), and FTO. The cardiac tissue was stained with Masson's trichrome and wheat germ agglutinin(WGA) to reveal the pathological changes. Immunohistochemistry was employed to detect the expression levels of Col1α, Col3α, α-SMA, and FTO in the myocardial tissue. The m6A modification level in the myocardial tissue was measured by the m6A assay kit. An H9c2 cell model of cardiomyocyte injury was induced by angiotensin Ⅱ(AngⅡ), and small interfering RNA(siRNA) was employed to knock down FTO expression. RT-qPCR was conducted to assess the relative m RNA levels of FTO and other genes associated with cardiac remodeling. The m6A modification level was measured by the m6A assay kit, and Western blot was employed to determine the phosphorylated phosphatidylinositol 3-kinase(p-PI3K)/phosphatidylinositol 3-kinase(PI3K) and phosphorylated serine/threonine kinase(p-Akt)/serine/threonine kinase(Akt) ratios in cardiomyocytes. The results of animal experiments showed that the XYP treatment significantly improved the cardiac function, reduced fibrosis, up-regulated the m RNA and protein levels of FTO, and lowered the m6A modification level compared with the model group. The results of cell experiments showed that the XYP-containing serum markedly up-regulated the m RNA level of FTO while decreasing the m6A modification level and the p-PI3K/PI3K and p-Akt/Akt ratios in cardiomyocytes. Furthermore, FTO knockdown reversed the protective effects of XYP-containing serum on Ang Ⅱ-induced cardiomyocyte hypertrophy. In conclusion, XYP may ameliorate ventricular remodeling by regulating the FTO/m6A axis, thereby inhibiting the activation of the PI3K/Akt signaling pathway.
Animals
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Ventricular Remodeling/drug effects*
;
Heart Failure/physiopathology*
;
Signal Transduction/drug effects*
;
Mice
;
Male
;
Alpha-Ketoglutarate-Dependent Dioxygenase FTO/genetics*
;
Drugs, Chinese Herbal/administration & dosage*
;
Mice, Inbred C57BL
;
Humans
;
Adenosine/analogs & derivatives*
;
Myocytes, Cardiac/metabolism*
;
Disease Models, Animal
5.Heart Yin deficiency and cardiac fibrosis: from pathological mechanisms to therapeutic strategies.
Jia-Hui CHEN ; Si-Jing LI ; Xiao-Jiao ZHANG ; Zi-Ru LI ; Xing-Ling HE ; Xing-Ling CHEN ; Tao-Chun YE ; Zhi-Ying LIU ; Hui-Li LIAO ; Lu LU ; Zhong-Qi YANG ; Shi-Hao NI
China Journal of Chinese Materia Medica 2025;50(7):1987-1993
Cardiac fibrosis(CF) is a cardiac pathological process characterized by excessive deposition of extracellular matrix(ECM). When the heart is damaged by adverse stimuli, cardiac fibroblasts are activated and secrete a large amount of ECM, leading to changes in cardiac fibrosis, myocardial stiffness, and cardiac function declines and accelerating the development of heart failure. There is a close relationship between heart yin deficiency and cardiac fibrosis, which have similar pathogenic mechanisms. Heart Yin deficiency, characterized by insufficient Yin fluids, causes the heart to lose its nourishing function, which acts as the initiating factor for myocardial dystrophy. The deficiency of body fluids leads to stagnation of blood flow, resulting in blood stasis and water retention. Blood stasis and water retention accumulate in the heart, which aligns with the pathological manifestation of excessive deposition of ECM, as a tangible pathogenic factor. This is an inevitable stage of the disease process. The lingering of blood stasis combined with water retention eventually leads to the generation of heat and toxins, triggering inflammatory responses similar to heat toxins, which continuously stimulate the heart and cause the ultimate outcome of CF. Considering the syndrome of heart Yin deficiency, traditional Chinese medicine capable of nourishing Yin, activating blood, and promoting urination can reduce myocardial cell apoptosis, inhibit fibroblast activation, and lower the inflammation level, showing significant advantages in combating CF.
Humans
;
Fibrosis/drug therapy*
;
Animals
;
Yin Deficiency/metabolism*
;
Myocardium/metabolism*
;
Medicine, Chinese Traditional
;
Drugs, Chinese Herbal/therapeutic use*
6.Efficacy and mechanism of Guizhi Tongluo Tablets in alleviating atherosclerosis by inhibiting CD72hi macrophages.
Xing-Ling HE ; Si-Jing LI ; Zi-Ru LI ; Dong-Hua LIU ; Xiao-Jiao ZHANG ; Huan HE ; Xiao-Ming DONG ; Wen-Jie LONG ; Wei-Wei ZHANG ; Hui-Li LIAO ; Lu LU ; Zhong-Qi YANG ; Shi-Hao NI
China Journal of Chinese Materia Medica 2025;50(5):1298-1309
This study investigates the effect and underlying mechanism of Guizhi Tongluo Tablets(GZTL) in treating atherosclerosis(AS) in a mouse model. Apolipoprotein E-knockout(ApoE~(-/-)) mice were randomly assigned to the following groups: model, high-, medium-, and low-dose GZTL, and atorvastatin(ATV), and age-matched C57BL/6J mice were selected as the control group. ApoE~(-/-) mice in other groups except the control group were fed with a high-fat diet for the modeling of AS and administrated with corresponding drugs via gavage for 8 weeks. General conditions, signs of blood stasis, and body mass of mice were monitored. Aortic plaques and their stability were assessed by hematoxylin-eosin, Masson, and oil red O staining. Serum levels of total cholesterol(TC), triglycerides(TG), and low-density lipoprotein cholesterol(LDL-C) were measured by biochemical assays, and those of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6) were determined via enzyme-linked immunosorbent assay. Apoptosis was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL). Single-cell RNA sequencing(scRNA-seq) was employed to analyze the differential expression of CD72hi macrophages(CD72hi-Mφ) in the aortas of AS patients and mice. The immunofluorescence assay was employed to visualize CD72hi-Mφ expression in mouse aortic plaques, and real-time fluorescence quantitative PCR was utilized to determine the mRNA levels of IL-1β, TNF-α, and IL-6 in the aorta. The results demonstrated that compared with the control group, the model group exhibited significant increases in body mass, aortic plaque area proportion, necrotic core area proportion, and lipid deposition, a notable decrease in collagen fiber content, and an increase in apoptosis. Additionally, the model group showcased elevated serum levels of TC, TG, LDL-C, IL-1β, TNF-α, and IL-6, alongside marked upregulations in the mRNA levels of IL-1β, TNF-α, and IL-6 in the aorta. In comparison with the model group, the GZTL groups and the ATV group showed a reduction in body mass, and the medium-and high-dose GZTL groups and the ATV group demonstrated reductions in aortic plaque area proportion, necrotic core area proportion, and lipid deposition, an increase in collagen fiber content, and a decrease in apoptosis. Furthermore, the treatment goups showcased lowered serum levels of TC, TG, LDL-C, IL-1β, TNF-α, and IL-6. The data of scRNA-seq revealed significantly elevated CD72hi-Mφ signaling in carotid plaques of AS patients compared with that in the normal arterial tissue. Animal experiments confirmed that CD72hi-Mφ expression, along with several pro-inflammatory cytokines, was significantly upregulated in the aortas of AS mice, which were downregulated by GZTL treatment. In conclusion, GZTL may alleviate AS by inhibiting CD72hi-Mφ activity.
Animals
;
Drugs, Chinese Herbal/administration & dosage*
;
Atherosclerosis/immunology*
;
Mice
;
Mice, Inbred C57BL
;
Macrophages/immunology*
;
Male
;
Humans
;
Apolipoproteins E/genetics*
;
Tablets
;
Tumor Necrosis Factor-alpha/genetics*
;
Apoptosis/drug effects*
;
Interleukin-1beta/genetics*
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Interleukin-6/genetics*
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Disease Models, Animal
;
Mice, Knockout
7.Mechanism of Qingrun Decoction in alleviating hepatic insulin resistance in type 2 diabetic rats based on amino acid metabolism reprogramming pathways.
Xiang-Wei BU ; Xiao-Hui HAO ; Run-Yun ZHANG ; Mei-Zhen ZHANG ; Ze WANG ; Hao-Shuo WANG ; Jie WANG ; Qing NI ; Lan LIN
China Journal of Chinese Materia Medica 2025;50(12):3377-3388
This study aims to investigate the mechanism of Qingrun Decoction in alleviating hepatic insulin resistance in type 2 diabetes mellitus(T2DM) rats through the reprogramming of amino acid metabolism. A T2DM rat model was established by inducing insulin resistance through a high-fat diet combined with intraperitoneal injection of streptozotocin. The model rats were randomly divided into five groups: model group, high-, medium-, and low-dose Qingrun Decoction groups, and metformin group. A normal control group was also established. The rats in the normal and model groups received 10 mL·kg~(-1) distilled water daily by gavage. The metformin group received 150 mg·kg~(-1) metformin suspension by gavage, and the Qingrun Decoction groups received 11.2, 5.6, and 2.8 g·kg~(-1) Qingrun Decoction by gavage for 8 weeks. Blood lipid levels were measured in different groups of rats. Pathological damage in rat liver tissue was assessed by hematoxylin-eosin(HE) staining and oil red O staining. Transcriptome sequencing and untargeted metabolomics were performed on rat liver and serum samples, integrated with bioinformatics analyses. Key metabolites(branched-chain amino acids, BCAAs), amino acid transporters, amino acid metabolites, critical enzymes for amino acid metabolism, resistin, adiponectin(ADPN), and mammalian target of rapamycin(mTOR) pathway-related molecules were quantified using quantitative real-time polymerase chain reaction(qRT-PCR), Western blot, and enzyme-linked immunosorbent assay(ELISA). The results showed that compared with the normal group, the model group had significantly increased serum levels of total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), and resistin and significantly decreased ADPN levels. Hepatocytes in the model group exhibited loose arrangement, significant lipid accumulation, fatty degeneration, and pronounced inflammatory cell infiltration. In liver tissue, the mRNA transcriptional levels of solute carrier family 7 member 2(Slc7a2), solute carrier family 38 member 2(Slc38a2), solute carrier family 38 member 4(Slc38a4), and arginase(ARG) were significantly downregulated, while the mRNA transcriptional levels of solute carrier family 1 member 4(Slc1a4), solute carrier family 16 member 1(Slc16a1), and methionine adenosyltransferase(MAT) were upregulated. Furthermore, the mRNA transcription and protein expression levels of branched-chain α-keto acid dehydrogenase E1α(BCKDHA) and DEP domain-containing mTOR-interacting protein(DEPTOR) were downregulated, while mRNA transcription and protein expression levels of mTOR, as well as ribosomal protein S6 kinase 1(S6K1), were upregulated. The levels of BCAAs and S-adenosyl-L-methionine(SAM) were elevated. The serum level of 6-hydroxymelatonin was significantly reduced, while imidazole-4-one-5-propionic acid and N-(5-phospho-D-ribosyl)anthranilic acid levels were significantly increased. Compared with the model group, Qingrun Decoction significantly reduced blood lipid and resistin levels while increasing ADPN levels. Hepatocytes had improved morphology with reduced inflammatory cells, and fatty degeneration and lipid deposition were alleviated. Differentially expressed genes and differential metabolites were mainly enriched in amino acid metabolic pathways. The expression levels of Slc7a2, Slc38a2, Slc38a4, and ARG in the liver tissue were significantly upregulated, while Slc1a4, Slc16a1, and MAT expression levels were significantly downregulated. BCKDHA and DEPTOR expression levels were upregulated, while mTOR and S6K1 expression levels were downregulated. Additionally, the levels of BCAAs and SAM were significantly decreased. The serum level of 6-hydroxymelatonin was increased, while those of imidazole-4-one-5-propionic acid and N-(5-phospho-D-ribosyl)anthranilic acid were decreased. In summary, Qingrun Decoction may improve amino acid metabolism reprogramming, inhibit mTOR pathway activation, alleviate insulin resistance in the liver, and mitigate pathological damage of liver tissue in T2DM rats by downregulating hepatic BCAAs and SAM and regulating key enzymes involved in amino acid metabolism, such as BCKDHA, ARG, and MAT, as well as amino acid metabolites and transporters.
Animals
;
Drugs, Chinese Herbal/administration & dosage*
;
Rats
;
Insulin Resistance
;
Diabetes Mellitus, Type 2/genetics*
;
Male
;
Liver/drug effects*
;
Amino Acids/metabolism*
;
Rats, Sprague-Dawley
;
Humans
;
Metabolic Reprogramming
8.Professor YANG Zhong-qi's prescription patterns for hypertension based on latent structure model and association rule analysis.
Hui-Lin LIU ; Shi-Hao NI ; Xiao-Jiao ZHANG ; Wen-Jie LONG ; Xiao-Ming DONG ; Zhi-Ying LIU ; Hui-Li LIAO ; Zhong-Qi YANG
China Journal of Chinese Materia Medica 2025;50(10):2865-2874
Based on latent structure model and association rule analysis, this study investigates the prescription patterns used by professor YANG Zhong-qi in treating hypertension with traditional Chinese medicine(TCM) and infers the associated TCM syndromes, providing a reference for clinical syndrome differentiation and treatment. The observation window spanned from January 8, 2013, to June 26, 2024, during which qualified herbal decoction prescriptions meeting efficacy criteria were extracted from the outpatient medical record system of the First Affiliated Hospital of Guangzhou University of Chinese Medicine and compiled into a standardized database. Statistical analysis of high-frequency herbs included frequency counts and herbal property-channel tropism analysis. Latent structure modeling and association rule analysis were performed using R 4.3.2 and Lantern 5.0 software to identify core herbal combinations and infer TCM syndrome patterns. A total of 2 436 TCM prescriptions were included in the study, involving 263 drugs with a cumulative frequency of 29 783. High-frequency herbs comprised Uncariae Ramulus cum Uncis, Poria, Glycyrrhizae Radix et Rhizoma, Puerariae Lobatae Radix, and Alismatis Rhizoma, predominantly categorized as deficiency-tonifying, heat-clearing, and blood-activating and stasis-resolving herbs. Latent structure analysis identified 18 latent variables, 74 latent classes, 5 comprehensive clustering models, and 15 core herbal combinations, suggesting that the core syndrome clusters include liver Yang hyperactivity pattern, Yin deficiency with Yang hyperactivity pattern, phlegm-stasis intermingling pattern, and liver-kidney insufficiency pattern. Association rule analysis revealed 22 robust association rules. RESULTS:: indicate that hypertension manifests as a deficiency-rooted excess manifestation, significantly associated with functional dysregulation of the liver, lung, spleen-stomach, heart, and kidney. Key pathogenic mechanisms involve liver Yang hyperactivity, phlegm-stasis interaction, and liver-kidney insufficiency. Therapeutic strategies should prioritize liver-calming, spleen-fortifying, and deficiency-tonifying principles, supplemented by dynamic regulation of Qi-blood and Yin-Yang balance according to syndrome evolution, alongside pathogen-eliminating methods such as phlegm-resolving and stasis-dispelling. Synergistic interventions like mind-tranquilizing therapies should be tailored to individual conditions.
Hypertension/drug therapy*
;
Drugs, Chinese Herbal/therapeutic use*
;
Humans
;
Medicine, Chinese Traditional
;
Drug Prescriptions
;
Latent Class Analysis
9.Application of active glucose monitoring in the perioperative period of gastrointestinal endoscopy in children with glycogen storage disease type Ⅰb.
Jing YANG ; Hao-Tian WU ; Ni MA ; Jia-Xing WU ; Min YANG
Chinese Journal of Contemporary Pediatrics 2025;27(8):923-928
OBJECTIVES:
To investigate the role of active glucose monitoring in preventing hypoglycemia during the perioperative period of gastrointestinal endoscopy in children with glycogen storage disease type Ⅰb (GSD-Ⅰb).
METHODS:
A retrospective analysis was performed for the clinical data of children with GSD-Ⅰb who were diagnosed and treated in Guangdong Provincial People's Hospital from June 2021 to August 2024. The effect of active glucose monitoring on hypoglycemic episodes during the perioperative period of gastrointestinal endoscopy was analyzed.
RESULTS:
A total of 14 children with GSD-Ⅰb were included, among whom there were 7 boys and 7 girls, with a mean age of 10.0 years. Among 34 hospitalizations, there were 15 cases of hypoglycemic episodes (44%), among which 6 symptomatic cases (1 case with blood glucose level of 1.6 mmol/L and 5 cases with blood glucose level of <1.1 mmol/L) occurred without active monitoring, while 9 asymptomatic cases (with blood glucose level of 1.2-3.9 mmol/L) were detected by active monitoring. The predisposing factors for hypoglycemic episodes included preoperative fasting (5 cases, 33%), delayed feeding (7 cases, 47%), vomiting (2 cases, 13%), and parental omission (1 case, 7%). Two children experienced two hypoglycemic episodes during the same period of hospitalization, and no child experienced subjective symptoms prior to hypoglycemic episodes. Treatment methods included nasogastric glucose administration (1 case, 7%), intravenous injection of glucose (14 cases, 93%), and continuous glucose infusion (4 cases, 27%). Blood glucose returned to 3.5-6.9 mmol/L within 10 minutes after intervention and remained normal after dietary resumption.
CONCLUSIONS
Active glucose monitoring during the perioperative period of gastrointestinal endoscopy can help to achieve early detection of hypoglycemic states in children with GSD-Ⅰb, prevent hypoglycemic episodes, and enhance precise diagnosis and treatment.
Humans
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Female
;
Male
;
Child
;
Retrospective Studies
;
Blood Glucose/analysis*
;
Hypoglycemia/etiology*
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Glycogen Storage Disease Type I/blood*
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Endoscopy, Gastrointestinal
;
Perioperative Period
;
Child, Preschool
;
Adolescent
10.Trace component fishing strategy based on offline two-dimensional liquid chromatography combined with PRDX3-surface plasmon resonance for Uncaria alkaloids.
Hui NI ; Zijia ZHANG ; Ye LU ; Yaowen LIU ; Yang ZHOU ; Wenyong WU ; Xinqin KONG ; Liling SHEN ; Sihan CHEN ; Huali LONG ; Cheng LUO ; Hao ZHANG ; Jinjun HOU ; Wanying WU
Journal of Pharmaceutical Analysis 2025;15(9):101244-101244
The rapid screening of bioactive constituents within traditional Chinese medicine (TCM) presents a significant challenge to researchers. Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents. To address this limitation, a fishing strategy based on offline two-dimensional liquid chromatography (2D-LC) combined with surface plasmon resonance (SPR) was utilized to screen bioactive trace components targeting peroxiredoxin 3 (PRDX3), using Uncaria alkaloids (UAs) as a case study. Initially, an orthogonal preparative offline 2D-LC system combining a positively charged C18 column and a conventional C18 column under disparate mobile phase conditions was constructed. To fully reveal the trace alkaloids, 13 2D fractions of UAs were prepared, and their components were characterized using mass spectrometry (MS). Subsequently, employing PRDX3 as the targeting protein, a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether (GSM) serving as a positive control for binding. Employing this refined strategy, 29 candidate binding alkaloids were fished from the 13 2D fractions. Notably, combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone. Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids, with isovallesiachotamine (IV), corynoxeine N-oxide (CO-N), and cadambine (CAD) demonstrating the highest affinity for PRDX3. Their interactions were further validated through molecular docking analysis. Subsequent intracellular H2O2 measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H2O2 clearance. In conclusion, this study introduced an innovative strategy for the identification of active trace components in TCM. This approach holds promise for accelerating research on medicinal components within this field.

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