1.Systematic review of risk prediction models for chemotherapy-induced myelosuppression in pediatric patients with malignant tumors
Li HE ; Xin LIN ; Xiaoping JIANG
China Pharmacy 2026;37(7):954-959
OBJECTIVE To systematically evaluate risk prediction models for chemotherapy-induced myelosuppression in pediatric patients with malignant tumors, evaluate their modeling strategies, key predictors, and predictive performance, and provide evidence-based references for clinical decision-making and research. METHODS A literature search was conducted across 11 databases, including CNKI, Wanfang Data, and PubMed, for relevant studies published before April 2025. Two reviewers independently performed literature screening and data extraction, and the risk of bias and applicability of the models were evaluated using the PROBAST tool. RESULTS Ultimately, seven studies were selected, of which four were English articles and three were Chinese articles, involving 12 risk prediction models. Although model discrimination was good (AUC 0.748-0.981), only two models underwent external validation; furthermore, calibration was inadequately reported in three studies. PROBAST indicated that all models exhibited a high risk of bias, with major issues including a predominance of retrospective designs, inadequate sample representativeness, and lack of blinding. However, in terms of applicability, all models received favorable evaluations. In terms of modeling methods, most studies employed traditional logistic regression approaches to construct models, while only a minority introduced machine learning algorithms and conducted systematic comparisons among multiple algorithms. Models developed using machine learning methods significantly outperformed those constructed with traditional statistical methods. CONCLUSIONS The existing risk prediction models for myelosuppression after chemotherapy in children with malignant tumors demonstrate potential in clinical risk early warning. However, they generally suffer from design and methodological limitations, such as a predominance of retrospective single-center designs, few events per variable, opaque handling of missing data, and inconsistent reporting of model coefficients. Future studies should adopt prospective designs, incorporate machine learning with key clinical predictors, and follow TRIPOD reporting guidelines to enhance scientific rigor and clinical utility.
2.Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of Periplaneta americana Extract Against Lung Adenocarcinoma
Qiong MA ; Chunxia HUANG ; Jiawei HE ; Yuting BAI ; Xingyue LIU ; Yuxuan XIONG ; Yang ZHONG ; Hengzhou LAI ; Yuling JIANG ; Xueke LI ; Qian WANG ; Yifeng REN ; Xi FU ; Funeng GENG ; Taoqing WU ; Ping XIAO ; Fengming YOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):124-132
ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract(PAE) against human-derived lung adenocarcinoma organoids(LUAD-PDOs) and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid(Nor-PDOs) models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8(CCK-8) assay. Experimental groups included the model group(LUAD-PDOs), normal group, model administration group(LUAD-PDOs+PAE), and normal administration group(Nor-PDOs+PAE). Hematoxylin-eosin(HE) staining was used to observe the pathological structures of PDOs, immunohistochemistry(IHC) was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7(CK-7) and Napsin A, TUNEL staining was applied to detect cell apoptosis. RNA sequencing(RNA-Seq) was conducted to identify differentially expressed genes(DEGs), followed by Gene Ontology(GO), Kyoto Encyclopedia of Genes and Genomes(KEGG), and Gene Set Enrichment Analysis(GSEA), alongside protein-protein interaction(PPI) network analysis to screen core mechanisms. Finally, key targets were validated by integrating external database analysis with immunofluorescence(IF). ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures, whereas LUAD-PDOs displayed dense, solid structures. CCK-8 and TUNEL assays revealed that, compared with the model group, PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells, while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention(347 up-regulated and 372 down-regulated)(P<0.05). GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism, including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway(P<0.05). PPI network analysis indicated that breast cancer type 1 susceptibility protein(BRCA1) and checkpoint kinase 1(CHEK1) were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention(P<0.05). Furthermore, survival analysis based on The Cancer Genome Atlas(TCGA) database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD(P<0.05). ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis, its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway, with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.
3.From Cathartic Colon to Cathartic-dependent Constipation: Diagnostic-therapeutic Strategies from Integrative Medicine Perspective
Youcheng HE ; Fengru JIANG ; Yanru WANG ; Minghan HUANG ; Yue WU ; Chunyu ZHOU ; Lian MO ; Lifeng WEI ; Keyi PAN ; Shuyu CAI ; Jianye YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):162-172
Both cathartic colon (CC) and cathartic-dependent constipation (CDC) are caused by the abuse of stimulant laxatives, while their concepts are not completely the same.Starting from the disease name of CC, this article traced the origin and evolution of the concept of CC, summarizes and compared the similarities and differences between CC, CDC, and slow transit constipation (STC), and called for strict differentiation among the three.Furthermore, this article explored the specific contents of Western medicine clinical subtypes and traditional Chinese medicine (TCM) syndrome differentiation of CDC and delved into the TCM pathogenesis of CDC according to both literature and clinical practice.The relationship between clinical subtypes and TCM syndromes was established, and the syndrome characteristics of CDC of different clinical subtypes and TCM syndromes were summarized.The recommended prescriptions for corresponding syndromes were listed.A systematic CDC diagnosis and treatment approach of "clinical subtypes-syndrome differentiation-syndrome characteristics-recommended prescriptions" was thus formed.Additionally, the paper provides an overview of current research on CDC in both Western medicine and TCM contexts, identifies future research directions, and suggests research pathways for refining and advancing CDC studies.
4.Clinical Efficacy of Yiqi Yangyin Huoxue Prescription in Treatment of Cathartic Colon and Analysis of Influencing Factors of Disease Severity
Youcheng HE ; Jingyi SHAN ; Fengru JIANG ; Yue WU ; Chunyu ZHOU ; Lu HANG ; Yan ZHOU ; Lian MO ; Shuyu CAI ; Keyi PAN ; Lifeng WEI ; Jianye YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):173-184
ObjectiveTo observe the clinical efficacy of the Yiqi Yangyin Huoxue prescription (YYHP) in the treatment of cathartic colon (CC) and its effects on fecal short-chain fatty acids (SCFAs), and to explore the correlations among CC severity indicators and between these indicators and patient history. MethodsAccording to the inclusion and exclusion criteria, 98 patients meeting the diagnostic criteria of both traditional Chinese and Western medicine for CC with the syndrome of Qi-Yin deficiency complicated by blood stasis were randomly assigned to an observation group and a control group. The observation group received YYHP granules, while the control group received lactulose. Both medications were administered twice daily, one sachet each time, half an hour after breakfast and dinner, with a treatment course of 8 weeks. The primary constipation symptom score, Patient Assessment of Constipation Quality of Life (PAC-QOL) score, and TCM syndrome score were assessed before and after treatment and at the 8th week after the end of treatment. The overall clinical effective rate, as well as the efficacy attenuation index and degree, were evaluated. Fecal SCFA levels were measured using gas chromatography-mass spectrometry (GC-MS). Spearman correlation analysis was performed to explore the correlations among CC severity indicators and between these indicators and patient history. ResultsThe overall clinical effective rate in the observation group (95.83%) was higher than that in the control group (78.72%) (P<0.05). After treatment, the total scores for primary constipation symptoms, PAC-QOL, and TCM syndromes decreased in both groups (P<0.05), with more significant reductions in the observation group (P<0.05). The severity of all primary constipation symptoms was alleviated in both groups (P<0.05). In terms of "excessive straining and difficult defecation", "anal heaviness, incomplete evacuation, and bloating sensation", "abdominal distension", and "defecation frequency", the observation group showed better efficacy than the control group (P<0.05). Scores of the four PAC-QOL dimensions and the scores and severity of primary and secondary TCM symptoms were reduced in both groups (P<0.05), with more significant reductions in the observation group (P<0.05). After treatment, acetic acid, propionic acid, butyric acid, and total SCFAs in the observation group increased significantly (P<0.05). The efficacy attenuation index and degree in the observation group were lower than those in the control group (P<0.05). No severe adverse reactions occurred in either group, and there was no statistically significant difference in the incidence of adverse reactions between the two groups. Positive correlations of varying degrees were observed among the total scores of primary constipation symptoms, PAC-QOL, and TCM syndromes, as well as between these scores and the history of stimulant laxative use, disease duration, and age. ConclusionYYHP can effectively alleviate the primary constipation symptoms in CC patients, improve quality of life, and ameliorate TCM syndromes, with good safety. It also has the advantage of a lower rebound degree after drug withdrawal, and its mechanism may be related to increasing fecal SCFA levels. Long-term abuse of stimulant laxatives may aggravate the severity of CC and prolong the disease course.
5.Qi and Yin Deficiency with Qi Stagnation and Blood Stasis in Rat Model of Rhein-induced Cathartic Colon
Youcheng HE ; Fengru JIANG ; Yue WU ; Chunyu ZHOU ; Erhao LIU ; Pengfei ZHOU ; Keyi PAN ; Shuyu CAI ; Jianye YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):185-195
ObjectiveTo evaluate and analyze the syndrome characteristics of Qi and Yin deficiency accompanied by Qi stagnation and blood stasis in a rhein-induced cathartic colon (CC) rat model. MethodsTwenty-four rats were divided into a normal group and a model group (CC group). The rats were administered equal volumes of physiological saline or 2% rhein suspension by gavage to establish the model over three cycles (approximately 118 days). The first cycle lasted 46 days, with a dosage of 12 mL·kg-1·d-1, administered every other day. The second cycle lasted 37 days, with a dosage of 12 mL·kg-1·d-1, administered for 5 consecutive days followed by 2 days of cessation. The third cycle lasted 35 days, with a dosage of 16 mL·kg-1·d-1, also administered for 5 consecutive days followed by 2 days of cessation. Each cycle ended when 80% of the rats no longer exhibited loose stools. Body mass, 24 h food intake, coat condition, and coat red (R), green (G), and blue (B) values were recorded. The open field test (OFT) was used to measure the total distance traveled to evaluate Qi deficiency. The body mass coefficient and 24 h water intake were recorded to assess Yin deficiency. The sucrose preference test (SPT) was used to determine the sucrose preference rate (SPR), and the average speed in OFT was measured to evaluate depressive status (liver depression and Qi stagnation). Tongue images and their R, G, and B values were recorded. Whole blood viscosity (WBV) and plasma viscosity (PV) were measured using an automatic hemorheological analyzer to evaluate blood stasis. A carbon ink propulsion test was performed to determine the intestinal transit rate (ITR) for disease model evaluation. Hematoxylin-eosin (HE) staining was used to observe histopathological changes in the colon. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of transient receptor potential ankyrin 1 (TRPA1) and tryptophan hydroxylase 1 (TPH1) in colon tissue. Western blot was used to detect the protein expression of TRPA1 and TPH1. ResultsIn terms of syndrome indicators, compared with the normal group, the body mass of the CC group decreased (P<0.05), while 24 h food intake increased (P<0.01). The coats of the CC group appeared withered, disheveled, and dull, and the R, G, and B values of the coat decreased (P<0.01). The total distance traveled in OFT decreased (P<0.01). The body mass coefficient decreased (P<0.01), while 24 h water intake increased (P<0.05, P<0.01). The SPR decreased (P<0.01), and the average speed in OFT slowed (P<0.01). The tongue appeared dark red, and the R, G, and B values of tongue images decreased (P<0.01). WBV and PV increased (P<0.01). Regarding disease indicators, compared with the normal group, the ITR decreased in the CC group (P<0.01). Pathologically, HE staining showed necrosis and shedding of colonic mucosal epithelial cells, disruption of mucosal continuity, and infiltration of inflammatory cells in the lamina propria in the CC group. Semi-quantitative analysis showed increased HAI scores (P<0.05) and increased inflammatory cell counts and area proportion (P<0.05). In terms of molecular biological indicators, compared with the normal group, the mRNA and protein expression levels of TRPA1 and TPH1 in colon tissue decreased in the CC group (P<0.05, P<0.01). ConclusionThe rhein-induced CC rat model conforms to the traditional Chinese medicine syndrome characteristics of Qi and Yin deficiency accompanied by Qi stagnation and blood stasis.
6.Animal Models of Functional Constipation: A Review
Youcheng HE ; Shijin LIN ; Fengru JIANG ; Sihan LI ; Xiao KE ; Wenrong WANG ; Jianye YUAN ; Minghan HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):196-209
Functional constipation (FC) is a clinically common functional bowel disorder characterized by a protracted course and associations with various chronic disorders and psychological abnormalities. Although not life-threatening, FC significantly impairs patients' quality of life. FC subtypes include slow-transit constipation (STC), defecatory disorder (DD), and normal-transit constipation (NTC). The pathological mechanisms underlying FC have not been fully elucidated, and overall clinical efficacy remains unsatisfactory. Animal models of FC serve as essential tools for the study of disease mechanisms and the development of novel therapeutics. This article systematically reviews the current state of research on the animal models of FC and identifies that rodents, particularly rats and mice, are the most commonly used species. Dogs and pigs are also employed in complex intervention studies due to their physiological similarities to humans, though their use is limited by housing challenges and ethical considerations. Induction methods vary across different FC subtypes. STC models are primarily established with chemical agents such as loperamide or compound diphenoxylate. DD modeling often involves low-fiber diets combined with methylene blue injection or rectal narrowing. NTC modeling mainly relies on low-fiber dietary interventions. In addition, disease-syndrome combination models based on traditional Chinese medicine (TCM) theory have been developed, encompassing excess patterns such as heat accumulation, cold accumulation, and Qi stagnation, as well as deficiency patterns including Qi deficiency, blood deficiency, Yin deficiency, and Yang deficiency. These are achieved through an approach of disease model + syndrome induction, enabling the integration of mechanisms from both Western and TCM perspectives. Models are evaluated from two aspects: disease and syndrome manifestations (e.g., colonic transit, secretory function, and TCM syndrome indicators such as mental state and body weight) and disease mechanisms (e.g., enteric nervous system, interstitial cells of Cajal, smooth muscle cells, gut microbiota, and metabolites). However, current research still faces challenges such as poor consistency in some models, non-specific interference in mechanism interpretation, insufficient studies on NTC, and lack of TCM tongue and pulse diagnosis in evaluation. Future efforts should focus on optimizing model stability and specificity to provide a more reliable experimental basis for investigating the pathological mechanisms of FC and developing therapeutic agents.
7.Effect and Mechanism of Xiao Qinglongtang Against Right Ventricular Dysfunction in Rats with Pulmonary Arterial Hypertension Induced by Monocrotaline
Lei QI ; Huifei ZHANG ; Ling GONG ; Jifu HE ; Wenjing CHEN ; Weipin NIU ; Xiao LI ; Yuehua JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):11-19
ObjectiveThis study aimed to establish a monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) rat model to systematically evaluate the protective effect of Xiao Qinglongtang (XQLT) on right cardiac function in model rats and further elucidate the underlying regulatory mechanism. MethodsSixty male SD rats were randomly assigned to the normal group, model group, XQLT low-, medium-, and high-dose groups (XQLT-L/M/H), and the beraprost sodium tablet group (BST). Except for the normal group, rats in all other groups were given a single subcutaneous injection of MCT (60 mg·kg-1) to induce PAH. Three weeks after injection, rats in the XQLT-L/M/H groups were administered XQLT intragastrically at 3.07, 6.14, 12.28 g·kg-1·d-1, respectively. Rats in the BST group received beraprost sodium at 12.6 μg·kg-1·d-1, and rats in the model group received an equal volume of saline. All treatments lasted for 3 weeks. Right ventricular systolic pressure (RVSP) was measured by right ventricular catheterization. Cardiac function was assessed by echocardiography. The right ventricle was weighed to calculate the right ventricular hypertrophy index (RVHI). Hematoxylin-eosin (HE) staining, Masson staining, and transmission electron microscopy were used to observe myocardial morphology. Serum metabolomic changes were analyzed using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Data-independent acquisition (DIA) proteomics was used to detect differentially expressed (DE) proteins in the right ventricle, and Western blot was used to measure the expression of uncoupling protein 3 (UCP3), phosphatidylinositol 3-kinase catalytic subunit p110α (PIK3CA), L1 cell adhesion molecule (L1CAM), and quinone oxidoreductase (CRYZ). UPLC-MS/MS was used to analyze the chemical components of XQLT. ResultsCompared with the normal group, the model group showed significantly increased RVSP and RVHI (P<0.05), along with pathological changes in myocardial morphology. Compared with the model group, all XQLT-treated groups exhibited reductions in RVSP and RVHI as well as significant improvements in cardiac function and myocardial morphology. Among the XQLT groups, XQLT-M showed the most pronounced effects (P<0.05), comparable to the BST group. Serum metabolomics revealed 105 differential metabolites in the XQLT groups versus the model group [variable importance in projection (VIP) >1, P<0.05], including 58 upregulated and 47 downregulated metabolites. KEGG enrichment analysis indicated that XQLT intervention downregulated phenylalanine metabolism (P<0.01) and upregulated unsaturated fatty acid biosynthesis (P<0.05). Proteomics analysis showed that 982 DE proteins were identified in the MCT groups versus the normal group, including 455 upregulated and 527 downregulated proteins (|fold change (FC)| >1.3, P<0.05). Compared with the model group, 237 DE proteins were identified in the XQLT groups, including 124 upregulated and 113 downregulated proteins (|FC| >1.3, P<0.05), with 57 overlapping DE proteins. KEGG enrichment suggested that XQLT mainly modulated pathways related to mineral absorption, ribosomal biogenesis, peroxisomes, glycolysis/gluconeogenesis, spliceosomes, and thyroid hormone signaling. Western blot analysis showed that, compared with the model group, XQLT increased the expression of UCP3, PIK3CA, and L1CAM, while decreasing the expression of CRYZ (P<0.05). ConclusionXQLT exerts a protective effect on right heart function in MCT-induced PAH rats, and its mechanism is associated with maintaining myocardial homeostasis and alleviating right ventricular remodeling.
8.Clinical Observation on Prevention of Recurrence of Common Bile Duct Stones After ERCP with Yuyin Lidan Granules
Xiao WANG ; Yong FANG ; Cong HE ; Jiali ZHANG ; Meng YU ; Jing KONG ; Yi JIANG ; Chuanqi CHENG ; Xiaosu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):159-166
ObjectiveTo observe the clinical efficacy and safety of Yuyin Lidan granules (YYLD) in preventing the recurrence of common bile duct stones (CBDS) in patients with liver and gallbladder dampness-heat syndrome following endoscopic retrograde cholangiopancreatography (ERCP). MethodsThis randomized, parallel, controlled trial enrolled postoperative CBDS-ERCP patients who met the inclusion and exclusion criteria. Sixty-four patients were randomly assigned to an observation group or a control group, with 32 cases in each. Both groups received conventional Western medical treatment after ERCP, while the observation group additionally received YYLD for 8 weeks. The follow-up period lasted for 1 year. The efficacy indicators included bile bilirubin levels, traditional Chinese medicine (TCM) syndrome scores, clinical efficacy rate, pancreatitis and inflammation markers, postoperative liver function, and CBDS recurrence rate at 1-year follow-up, which were used to jointly evaluate the clinical efficacy and safety of both groups. ResultsA total of 56 patients completed the study and were included in the final analysis, i.e., 29 in the observation group and 27 in the control group. Baseline characteristics were comparable between the two groups. Compared with pre-treatment and with the control group after treatment, the bile bilirubin level in the observation group significantly decreased (P<0.05). After treatment, the clinical cure and marked improvement rates were higher in the observation group than in the control group, showing a statistically significant difference in overall clinical efficacy (P<0.05). Compared with pre-treatment, the primary and secondary symptoms in the observation group, as well as the primary symptom and the secondary symptom of nausea and vomiting in the control group (weeks 4 and 8), were significantly reduced (P<0.05). Compared with the control group after treatment, the observation group showed significant reductions in the primary symptom of loose stools/constipation (day 5 and week 4) and in three secondary symptoms, i.e., bitter taste and sticky dry mouth, abdominal distension and poor appetite (throughout the treatment period), and general heaviness and fatigue (day 5 and week 4), with statistical differences (P<0.05). Compared with pre-treatment, both groups showed decreased lipase and urinary amylase levels (P<0.05). However, no significant between-group differences were observed in pancreatitis or inflammation-related indices after treatment. Compared with pre-treatment, all liver function indicators in the observation group and alanine aminotransferase ( ALT ), γ-glutamyl transferase ( γ-GT ), alkaline phosphatase (ALP), and conjugated bilirubin in the control group significantly decreased at weeks 4 and 8 (P<0.05). Compared with the control group after treatment, only serum total bilirubin and unconjugated bilirubin were significantly reduced in the observation group during the treatment period (P<0.05). ConclusionYYLD combined with conventional Western medical treatment can effectively regulate bilirubin metabolism (in bile and serum), improve TCM clinical symptoms, and prevent CBDS recurrence after ERCP in patients with liver and gallbladder dampness-heat syndrome. This regimen is safe and effective and is worthy of further clinical research and promotion.
9.Retrospective analysis and grey zone verification of blood donors with anti-Treponema pallidum single reagent reactive results
Qing HE ; Feifei JIANG ; Zhichao CHEN ; Panpan WANG ; Yousheng KONG ; Suping LI
Chinese Journal of Blood Transfusion 2026;39(2):224-228
Objective: To explore the setting of gray zone of Treponema pallidum (TP) testing by retrospective analysis of blood donors with single reagent reactive anti-TP results, so as to improve blood utilization and supply safety. Methods: Blood samples were collected from 112 blood donors previously deferred due to single reagent reactive TP antibody results between January 2020 and December 2023, and subjected to dual ELISA reagents and TPPA test. The gray zone panel analysis was performed on the two ELISA reagents currently used in our department. The detection rate at each concentration of the gray zone panle was counted, and the corresponding concentrations for C
, C
, and C
and gray zone cut-off were calculated. Results: Among the 50 samples deferred by reagent 1, 19 were confirmed reactive and 31 non-reactive in supplementary testing. Among the 62 samples deferred by reagent 2, 12 were confirmed reactive and 50 non-reactive in supplementary testing. For reagent 1, the detection rate of was 56% for S/CO≥1 and 20% for 0.5≤S/CO<1, retrospectively. For reagent 2, the detection rate was 27% for S/CO≥1 and 12.5% for 0.5≤S/CO<1, retrospectively. The detection rate for S/CO≥1 was higher than those for 0.5≤S/CO<1 for both reagents. All the 112 samples were negative in TPPA test. The C
concentration of reagent 1 was 1.51 mIU/mL, and the concentration range of C
±20% was 1.21-1.81 mIU/mL. The C
concentration of reagent 2 was 1.45 mIU/mL, and the concentration range of C
±20% was 1.16-1.74 mIU/mL. The C
and C
concentration of both reagents were within the C
±20% range, suggesting that the gray zone cutoff for both Reagent 1 and Reagent 2 should be set at S/CO=0.8 (80% of the CO value). Conclusion: All anti-TP single reagent reactive samples with S/CO value within the gray zone was tested negative by TPPA. It is necessary to consider the rationality and necessity of establishing the gray zone, so as to ensure blood safety and improve the utilization rate of blood resources.
10.Mechanism of Jinyang Dingtong Plaster in Improving Peripheral Pain Sensitization and Synovial Fibrosis in Knee Osteoarthritis by Blocking Ion Channels of TRPs
Jinliang HE ; Lu ZHANG ; Shixin XING ; Xilu REN ; Jingxing JIANG ; Junfeng KANG ; Xuliang HAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):108-116
ObjectiveThis paper aims to investigate the mechanism of Jinyang Dingtong plaster in improving the peripheral pain sensitization and synovial fibrosis in rats with knee osteoarthritis (KOA) by blocking the ion channels of transient receptor potentials (TRPs). MethodsThe active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by using ultra-high performance liquid chromatography-electrospray ionization-quadrupole ion trap tandem mass spectrometry (UPLC-MS/MS) technology. A KOA rat model was established through intra-articular injection of monoiodoacetic acid. The rats were randomly divided into blank control group, KOA group, compound Nanxing Zhitong plaster Group, and Jinyang Dingtong plaster group, with eight rats per group. Among them, the rats in the compound Nanxing Zhitong plaster group and the Jinyang Dingtong plaster group were intervened with external application treatment. After the intervention period, the cold and mechanical stimulus pain thresholds of rats in each group were detected, and the transverse diameter of the knee joint was measured. The levels of inflammatory factors in the serum such as interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), nerve growth factor (NGF), and calcitonin gene-related peptide (CGRP) were determined by enzyme-linked immunosorbent assay (ELISA). Protein expression levels of transient receptor potential ankyrin 1 (TRPA1), transient receptor potential melastatin 8 (TRPM8), transient receptor potential vanilloid 1 (TRPV1), transient receptor potential vanilloid 4 (TRPV4), transforming growth factor-β (TGF-β), and vascular endothelial growth factor (VEGF) in synovial tissue were detected by Western blot. Histopathological changes in synovial tissue were observed by using hematoxylin and eosin (HE), Masson, and Sirius red staining, while the expression of type Ⅰ collagen and alpha-smooth muscle actin (α-SMA) was detected by multiplex immunofluorescence. ResultsA total of 35 active components in the transdermal absorption solution of Jinyang Dingtong plaster were identified by UPLC-MS/MS, including phenolic acids, flavonoids, quinones, alkaloids, terpenes, lignans, and coumarins. Among them, the constituents such as berberine, paeoniflorin, ferulic acid, and caffeic acid exhibit clear anti-inflammatory, analgesic, and anti-fibrotic pharmacological effects. Compared to the blank control group, rats in the KOA group showed a significant decrease in cold and mechanical stimuli pain thresholds (P<0.01). After 14 and 28 days of Jinyang Dingtong plaster intervention, the pain threshold in this group was significantly increased compared to that in KOA group (P<0.01), showing no significant difference from that in compound Nanxing Analgesic plaster group. Additionally, Jinyang Dingtong plaster reduced the levels of IL-1β, TNF-α, NGF, and CGRP in the serum of KOA rats (P<0.01), lowered the expression of TRPA1, TRPM8, TRPV1, TRPV4, TGF-β, and VEGF proteins in synovial tissue (P<0.01), improved synovial pathological damage in KOA rats, and significantly decreased fluorescence intensity of type Ⅰ collagen and α-SMA (P<0.01). ConclusionJinyang Dingtong plaster can improve the peripheral pain sensitization and synovial fibrosis in KOA rats by downregulating the expression of ion channels of TRPs and related inflammatory and fibrotic factors.

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