1.Epidemiological characteristics and influencing factors of severe fever with thrombocytopenia syndrome in Zhejiang Province
LÜ ; Jing ; XU Xinying ; QIAO Yingyi ; SHI Xinglong ; YUE Fang ; LIU Ying ; CHENG Chuanlong ; ZHANG Yuqi ; SUN Jimin ; LI Xiujun
Journal of Preventive Medicine 2026;38(1):10-14
Objective:
To analyze the epidemiological characteristics and influencing factors of severe fever with thrombocytopenia syndrome (SFTS) in Zhejiang Province from 2019 to 2023, so as to provide the reference for strengthening SFTS prevention and control.
Methods:
Data on laboratory-confirmed SFTS cases in Zhejiang Province from 2019 to 2023 were collected through the Infectious Disease Reporting Information System of Chinese Disease Prevention and Control Information System. Meteorological data, geographic environment and socioeconomic factors during the same period were collected from the fifth-generation European Centre for Medium-Range Weather Forecasts, Geospatial Data Cloud, and Zhejiang Statistical Yearbook, respectively. Descriptive epidemiological methods were used to analyze the epidemiological characteristics of SFTS from 2019 to 2023, and a Bayesian spatio-temporal model was constructed to analyze the influencing factors of SFTS incidence.
Results:
A total of 578 SFTS cases were reported in Zhejiang Province from 2019 to 2023, with an annual average incidence of 0.23/105. The peak period was from May to July, accounting for 52.60%. There were 309 males and 269 females, with a male-to-female ratio of 1.15∶1. The cases were mainly aged 50-<80 years, farmers, and in rural areas, accounting for 82.53%, 77.34%, and 75.43%, respectively. Taizhou City and Shaoxing City reported more SFTS cases, while Shaoxing City and Zhoushan City had higher annual average incidences of SFTS. The Bayesian spatio-temporal interaction model showed good goodness of fit. The results showed that mean temperature (RR=1.626, 95%CI: 1.111-2.378) and mean wind speed (RR=1.814, 95%CI: 1.321-2.492) were positively correlated with SFTS risk, while altitude (RR=0.432, 95%CI: 0.230-0.829) and population density (RR=0.443, 95%CI: 0.207-0.964) were negatively correlated with SFTS risk.
Conclusions
SFTS in Zhejiang Province peaks from May to July. Middle-aged and elderly people and farmers are high-risk populations. Taizhou City, Shaoxing City, and Zhoushan City are high-incidence areas. Mean temperature, mean wind speed, altitude, and population density can all affect the risk of SFTS incidence.
2.Proteomic Analysis of Danlou Tablet in Improving Platelet Function for Treating Coronary Heart Disease with Phlegm-stasis Intermingling Syndrome in Minipigs
Ziyan WANG ; Ying LI ; Aoao WANG ; Hongxu MENG ; Yue SHI ; Yanlei MA ; Guoyuan ZHANG ; Lei LI ; Jianxun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):41-53
ObjectiveThis paper aims to observe the role of Danlou tablet in treating coronary heart disease (CHD) with phlegm-stasis intermingling syndrome in minipigs by improving platelet function and explore the potential pharmacological mechanism of Danlou tablet in regulating platelet function by using proteomics technology. MethodsThirty Bama minipigs were randomly divided into a normal control group (6 pigs) and a high-fat diet group (24 pigs). After 2 weeks of high-fat diet feeding, the high-fat diet group was randomly subdivided into a model group, an atorvastatin group (1 mg·kg-1), and Danlou tablet groups (0.6 g·kg-1 and 0.3 g·kg-1). All groups continued to receive a high-fat diet for 8 weeks after the procedure. The normal control group was given a regular diet, underwent only coronary angiography, and did not receive an interventional injury procedure. The model group and each administration group were fed a high-fat diet. Two weeks later, they underwent a coronary angiography injury procedure. After the procedure, drugs were mixed into the feed every morning for 8 consecutive weeks, with the minipigs maintained on a continuous high-fat diet during this period. Quantitative proteomics technology was further used to study platelet proteins, and differential proteins were obtained by screening. Bioinformatics analysis was performed to analyze key regulatory proteins and biological pathways involved in the therapeutic effect of Danlou tablet on CHD with phlegm-stasis intermingling syndrome. ResultsCompared with the normal control group, the model group showed a significant increase in total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) of minipigs' serum (P<0.01), a significant shortening in prothrombin time of (PT) (P<0.01), a coagulation function index, and an increase in whole blood viscosity (P<0.01) and platelet aggregation rate (P<0.01). Moreover, the platelet morphology was altered, and the contents of endothelin-1 (ET-1) and nitric oxide (NO) were significantly increased (P<0.01). Hemodynamic parameters were obviously abnormal, including significantly decreased systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), and left ventricular maximal positive dp/dt (LV+dp/dtmax) (P<0.01). Left ventricular maximal negative dp/dt (LV-dp/dtmax) was significantly increased (P<0.01). Besides, there were myocardial cell hypertrophy, obvious edematous degeneration, massive interstitial inflammatory cell infiltration, high degree of fibrosis, and coronary endothelial atherosclerosis. TC and TG levels in minipigs' serum were significantly reduced in Danlou tablet groups with 0.6 g·kg-1 and 0.3 g·kg-1 (P<0.05, P<0.01), compared with those in the model group. LDL-C was decreased in the Danlou tablet group with 0.6 g·kg-1 (P<0.05). The whole blood viscosity under low and high shear conditions was significantly reduced in the Danlou tablet group with 0.6 g·kg-1 (P<0.05). In groups with all doses of Danlou tablet, maximum aggregation rate (MAR) and average aggregation rate (AAR) were significantly decreased (P<0.05, P<0.01), and platelets' morphological changes such as pseudopodia extension were reduced. ET-1 levels in the serum were significantly reduced. In the Danlou tablet group with 0.6 g·kg-1, NO level in the serum was reduced (P<0.05). In groups with all doses of Danlou tablet, DBP and MAP were significantly increased (P<0.05). In the Danlou tablet group with 0.6 g·kg-1, LVSP and LV+dp/dtmax were significantly increased (P<0.05, P<0.01), and LV-dp/dtmax was significantly decreased (P<0.05). In groups with all doses of Danlou tablet, edematous degeneration in myocardial tissue was milder, and coronary artery lesion degree was significantly alleviated. Compared with the normal control group, there were 94 differentially expressed proteins in the model group, including 81 up-regulated and 13 down-regulated proteins. Compared with the model group, the Danlou tablet group with 0.6 g·kg-1 showed 174 differentially expressed proteins, including 100 up-regulated and 74 down-regulated proteins. A total of 30 proteins were reversed after Danlou tablet intervention. Bioinformatics analysis revealed that its pharmacological mechanism may exert anti-platelet activation, aggregation, and adhesion effects through biological pathways such as regulation of actin cytoskeleton, platelet activation pathway, Fcγ receptor-mediated phagocytosis, as well as proteins such as growth factor receptor-bound protein 2 (GRB2), Ras-related C3 botulinum toxin substrate 2 (RAC2), RAC1, and heat shock protein 90 alpha family class A member 1 (HSP90AA1). ConclusionDanlou tablet can effectively reduce platelet activation and aggregation, exerting a good therapeutic effect on CHD with phlegm-stasis intermingling syndrome in minipigs. Its pharmacological mechanism may involve regulating biological pathways such as actin cytoskeleton and platelet activation pathway, as well as proteins like GRB2, RAC2, RAC1, and HSP90AA1, thereby exerting a pharmacological effect in anti-platelet activation, aggregation, and adhesion.
3.Effect and Mechanism of Schisandrae Chinensis Fructus Lignans on Behavior of Schizophrenic Mice
Jiaqi LI ; Xi CHEN ; Siwei WANG ; Qi WANG ; Yiting LIU ; Ziyan GUO ; Zilong LUN ; Chengyi ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):65-71
ObjectiveTo investigate the effects of Schisandrae Chinensis Fructus lignans on schizophrenia induced by dizocilpine maleate (MK-801) in mice and to clarify its mechanism. MethodsMale mice of 4-6 weeks old were randomized into blank, model, positive drug, and low-, medium-, and high-dose (40, 80, 160 mg·kg-1, respectively) Schisandrae Chinensis Fructus lignans groups. The blank group was administrated with distilled water, and the other groups were injected with 0.5 mg·kg-1 MK-801 to induce schizophrenia symptoms. Meanwhile, risperidone was injected at 0.2 mg·kg-1 in the positive drug group, and mice in the intervention groups were injected with corresponding drugs for 14 consecutive days. The behavioral changes of mice were observed by autonomous activity test, open field test, forced swimming test, and water maze test. The levels of dopamine (DA) and 5-hydroxytryptamine (5-HT) in the brain and tumor necrosis factor-α (TNF-α) and nuclear factor-κB (NF-κB) in peripheral blood were quantified by enzyme-linked immunosorbent assay (ELISA). The changes in the prefrontal lobe of mice were observed by hematoxylin-eosin staining, and the changes of the hippocampal tissue were observed by Nissl staining. The protein levels of silencing information regulatory factor 1 (SIRT1) and forkhead box protein O3a (FoxO3a) in the hippocampus of mice were determined by Western blot. ResultsCompared with the model group, low, medium, and high doses of Schisandrae Chinensis Fructus lignans reduced the total number of autonomous activities, total distance in the open field test, immobile time in the forced swimming test, and levels of TNF-α and NF-κB in peripheral blood (P<0.05), while increasing the number of platform crossings in the water maze test and DA and 5-HT levels in the brain tissue (P<0.05). Compared with the model group, risperidone and low, medium, and high doses of Schisandrae Chinensis Fructus lignans improve the neural cell morphology in the CA1 region, with full cells in neatly dense arrangement and exhibiting clear membrane boundary. Schisandrae Chinensis Fructus lignans inhibited the expression of SIRT 1 and FoxO3a in the hippocampus (P<0.05). ConclusionTo sum up, Schisandrae Chinensis Fructus lignans may improve the behavior of schizophrenic mice by activating the SIRT1/FoxO3a signaling pathway to exert neuroprotective effects.
4.Mechanism of Xiaozheng Zhitong Paste in Alleviating Bone Cancer Pain by Regulating Microglial Pyroptosis Based on PINK1/Parkin/NLRP3 Signaling Pathway
Lingyun WANG ; Guangda ZHENG ; Lu SHANG ; Juanxia REN ; Changlin LI ; Dongtao LI ; Haixiao LIU ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):80-90
ObjectiveThe paper aims to investigate the mechanism by which Xiaozheng Zhitong paste (XZP) alleviates bone cancer pain (BCP) through regulating the PTEN-induced putative kinase 1 (PINK1)/Parkin-mediated mitophagy-NOD-like receptor protein 3 (NLRP3) inflammasome pathway to suppress microglial pyroptosis. MethodsLipopolysaccharide (LPS) and LPS-adenosine triphosphate (ATP) were used to establish an inflammation and pyroptosis model in microglial cells. The cells were randomly divided into the following groups: control group, LPS group, LPS+low-dose XZP group, LPS+high-dose XZP group, LPS-ATP group, LPS-ATP+low-dose XZP group, LPS-ATP+high-dose XZP group, LPS-ATP+XZP group, and LPS-ATP+XZP+CsA group. Techniques including terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining, enzyme-linked immunosorbent assay (ELISA), Western blot, and confocal fluorescence staining were employed to assess the effects of XZP on microglial apoptosis, inflammatory cytokine release, inflammasome activation, pyroptosis, and mitophagy. ResultsIn vitro experiments showed that compared with the blank group, the LPS group exhibited significantly increased levels of microglial apoptosis and pro-inflammatory factors interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α)(P<0.01), along with significantly upregulated protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and phosphorylated nuclear factor-κB p65 (p-NF-κB p65) (P<0.01). Compared with the LPS group, the high-dose LPS-XZP group significantly reduced the level of apoptosis (P<0.01) and the content of the aforementioned pro-inflammatory factors (P<0.01). Both the low- and high-dose LPS-XZP groups dose-dependently downregulated the protein expression of iNOS, COX-2, and p-NF-κB p65 (P<0.05, P<0.01). Compared with the blank group, the LPS-ATP group showed significantly upregulated expression of pyroptosis-related proteins, including Caspase-1/pro-Caspase-1, N-terminal fragment of gasdermin D (GSDMD-N)/full-length gasdermin D (GSDMD-F), NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), IL-1β precursor (pro-IL-1β), and mature IL-1β (P<0.01). The levels of pyroptotic factors IL-1β and IL-18 were significantly elevated (P<0.01), and membrane pore formation and intracellular reactive oxygen species (ROS) levels were significantly increased (P<0.01). Compared with the LPS-ATP group, both the low- and high-dose LPS-ATP+XZP groups dose-dependently downregulated the expression of the aforementioned pyroptosis-related proteins (P<0.05, P<0.01). The low-dose LPS-ATP+XZP group reduced IL-1β levels (P<0.01), while the high-dose group reduced both IL-1β and IL-18 levels (P<0.01) Both the low- and high-dose LPS-ATP+XZP groups dose-dependently reduced membrane pore formation and intracellular ROS production (P<0.01). Compared with the blank group, the LPS-ATP group showed significantly reduced expression of mitophagy-related proteins PINK1 and Parkin, and a decreased ratio of microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ) to LC3Ⅰ(P<0.01), while p62 expression was significantly increased (P<0.01). Mitochondrial ROS levels were significantly enhanced (P<0.01). Compared with the LPS-ATP group, both the low- and high-dose LPS-ATP+XZP groups dose-dependently reversed the expression of these proteins (P<0.05, P<0.01) and reduced mitochondrial ROS levels (P<0.01). After treatment with the mitophagy inhibitor cyclosporin A (CsA), the beneficial effects of XZP on mitochondrial function and its inhibitory effects on pyroptosis-related protein expression were significantly reversed (P<0.05, P<0.01). ConclusionXZP reduces ROS levels by activating PINK1/Parkin-mediated mitophagy, thereby inhibiting NLRP3 inflammasome activation and microglial pyroptosis, which provides new molecular evidence for the mechanism by which XZP alleviates BCP.
5.Xiaozheng Zhitong Paste Relieves Bone Cancer Pain in Mice by Alleviating Activation of Microglia in Spinal Cord and Damage to Neurons via Blocking PAR2/NF-κB/NLRP3 Pathway
Guangda ZHENG ; Linghan MENG ; Lu SHANG ; Juanxia REN ; Dongtao LI ; Haixiao LIU ; Lingyun WANG ; Changlin LI ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):91-100
ObjectiveTo investigate the effects and underlying mechanisms of Xiaozheng Zhitong Paste (XZP) on bone cancer pain (BCP). MethodsThirty female BALB/c mice were randomly divided into five groups: a Sham group, a BCP group, a BCP+low-dose XZP group, a BCP+high-dose XZP group, and a BCP+high-dose XZP + protease-activated receptor 2 (PAR2) agonist GB-110 group. BCP mice model was constructed by injecting Lewis lung carcinoma cells into the femoral cavity of the right leg, which was followed by being treated with XZP for 21 d. After 21 d, the mice were sacrificed. Nissl staining was used to evaluate the survival of spinal cord neurons. Immunofluorescence staining was conducted to localize ionized calcium-binding adapter molecule 1 (Iba1) and neuronal nuclear antigen (NeuN) in spinal cord tissue, thereby assessing microglial activation and neuronal survival. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), transforming growth factor-β (TGF-β), interleukin-4 (IL-4), and interleukin-10 (IL-10) in spinal cord tissue. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect mRNA expression levels associated with M1/M2 polarization of microglia. Western blot analysis was performed to examine the expression of proteins related to microglial polarization as well as those involved in the PAR2/nuclear factor kappa B (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway in the spinal cord. ResultsCompared with the Sham group, the spinal cord neurons were damaged, the number of Nissl-positive spinal cord neurons in the spinal cord tissue was significantly reduced (P<0.01), and the rate of NeuN-positive cells was significantly decreased (P<0.01). The spinal cord microglia were activated, the inflammatory level of the spinal cord tissue was enhanced, and Iba1 staining was significantly enhanced (P<0.01). The levels of IL-1β, TNF-α, IL-6, TGF-β, IL-4 and IL-10 were significantly increased (P<0.01). The mRNA expressions of IL-1β, TNF-α and inducible nitric oxide synthase (iNOS) were significantly increased (P<0.01), and the expression of PAR2, NLRP3, ASC and NF-κB p65 proteins in the spinal cord tissue of the BCP mice was significantly enhanced (P<0.01). Compared with the BCP group, high-dose XZP treatment significantly increased the number of Nissl-positive spinal cord neurons in the BCP mice (P<0.01), significantly enhanced the rate of NeuN-positive cells in the spinal cord tissue, and significantly weakened Iba1 staining (P<0.01). In addition, the levels of IL-1β, TNF-α, and IL-6 were significantly decreased, while the levels of TGF-β, IL-4, and IL-10 were significantly increased (P<0.05, P<0.01). The mRNA expression levels of IL-1β, TNF-α, and iNOS were decreased, whereas those of cluster of differentiation 206 (CD206), arginase-1 (Arg-1), and YM1/2 were significantly increased (P<0.05, P<0.01). Low-dose and high-dose XZP treatment significantly decreased the expression of PAR2, NLRP3, ASC, and NF-κB p65 proteins in the spinal cord tissue (P<0.05, P<0.01). These effects could all be significantly eliminated by the PAR2 agonist GB-110. ConclusionXZP can mitigate BCP in mice, which may be achieved through blocking the activated PAR2/NF-κB/NLRP3 pathway.
6.Traditional Chinese Medicine for Cancer Pain Management: A Review
Lingyun WANG ; Guangda ZHENG ; Lu SHANG ; Juanxia REN ; Changlin LI ; Dongtao LI ; Haixiao LIU ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):114-123
Cancer pain is one of the most common complications in patients with malignant tumors, severely affecting their quality of life. Its pathogenesis involves complex interactions among the tumor microenvironment, peripheral sensitization, and central sensitization. The tumor microenvironment initiates peripheral pain sensitization by secreting algogenic mediators, activating ion channels and related receptor signaling pathways, driving abnormal osteoclast activation, and mediating neuro-immune crosstalk. Persistent nociceptive input further triggers increased excitability of central neurons, activation of glial cells, and neuroinflammatory cascade reactions, ultimately leading to central pain sensitization. Although traditional opioid drugs can alleviate pain to some extent, they still have many limitations, such as incomplete analgesia, drug tolerance, and adverse reactions. In recent years, traditional Chinese medicine (TCM) compounds have made continuous progress in the treatment of cancer pain. Studies have shown that they can not only effectively relieve cancer pain and reduce the dosage of opioids but also significantly improve patients' quality of life. TCM treatment of cancer pain follows the principle of syndrome differentiation and treatment. Based on this, targeted therapeutic principles have been proposed, including promoting blood circulation, removing stasis, regulating Qi, and unblocking collaterals; tonifying the kidney, replenishing essence, warming Yang, and dispersing cold, activating blood, resolving phlegm, detoxifying, and dispersing nodules, as well as strengthening the body, replenishing deficiency, and harmonizing Qi and blood. Modern research indicates that TCM compounds can exert synergistic effects through multiple pathways, inhibiting inflammatory responses, regulating nerve conduction, intervening in bone metabolism and related gene expression, thereby producing anti-inflammatory and bone-protective effects to achieve the goal of alleviating cancer pain. This article systematically elaborates on the pathogenesis of cancer pain, the clinical application of TCM in treating cancer pain, and its related mechanisms of action, aiming to provide a theoretical basis and new strategies for the integration of TCM into comprehensive cancer pain management.
7.Discussion on Treatment of Cancer Pain with Modified Wumeiwan Based on Jueyin Syndrome
Haixiao LIU ; Linghan MENG ; Guangda ZHENG ; Dongtao LI ; Lu SHANG ; Juanxia REN ; Changlin LI ; Lingyun WANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):124-128
Pain, as one of the most common symptoms in cancer patients, seriously affects the survival quality of patients. The three-step pain relief program currently used in clinical practice cannot completely relieve pain in cancer patients and is accompanied by many problems. From the perspective of Jueyin syndrome in traditional Chinese medicine (TCM), this paper believed that the core pathogenesis of cancer pain was declined healthy Qi and cold and heat in complexity, and used Wumeiwan as the main formula with modification according to syndrome for clearing the upper, warming the lower part of the body, and harmonizing the cold and heat. It can regulate the pathological environment of deficiency, cold, stasis, toxicity, and heat, and restore the physiological function of Yang transforming Qi while Yin constituting form, so as to prevent, relieve, and even eliminate cancer pain, having achieved good clinical efficacy. It can not only help cancer patients relieve pain, but also control tumor and eliminate tumor, achieving a dual benefit of pain relief and tumor suppression. It gives full play to the characteristics and advantages of syndrome differentiation and treatment in TCM, and expands the scope of ZHANG Zhongjing's treatment for Jueyin syndrome, which provides ideas for the clinical diagnosis and treatment of cancer pain from the perspective of deficiency-excess in complexity and cold and heat in complexity.
8.Comparison of Wild and Cultivated Bupleurum scorzonerifolium Based on Traditional Quality Evaluation
Changsheng YUAN ; Feng ZHOU ; Xingyu LIU ; Yu SHI ; Yihan WANG ; Huaizhu LI ; Yongliang LI ; Shan GUAN ; Huaizhong GAO ; Yanmeng LIU ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):203-214
ObjectiveTo characterize the quality differences among different germplasm and introduced varieties of Bupleurum scorzonerifolium roots(BSR), and explore the underlying molecular mechanisms, providing a basis for high-quality production and quality control. MethodsWild BSR from Yulin(YLW) served as the quality reference, we conducted comparative analysis among YLW, locally domesticated wild germplasm in Yulin(YLC3), Daqing germplasm introduced and cultivated in Yulin(YLDQC3), and locally cultivated germplasm in Daqing(DQC3). A combination of traditional pharmacognostic methods and modern multi-omics analyses was employed, including macroscopic traits(appearance, odor), microscopic features(proportions of cork, phloem, xylem), cell wall component contents(hemicellulose, cellulose, lignin), carbohydrate contents(starch, water-soluble polysaccharides), marker compound contents(ethanol-soluble extracts, total saponins, liposoluble extracts, and saikosaponins A, B2, C, D), metabolomics, and transcriptomics, in order to systematically characterize quality differences and investigate molecular mechanisms among these samples. ResultsMacroscopically, Yulin-produced BSR(YLW, YLC3, YLDQC3) exhibited significantly greater weight, length, and upper and middle diameters than Daqing-produced BSR(DQC3). Odor-wise, YLW and YLC3 had a a fragrance taste, YLDQC3 had a rancid oil odor, and DQC3 had a sweet and fragrant taste. Microscopically, Yulin germplasm(YLW, YLC3) and Daqing germplasm(YLDQC3, DQC3) shared similar structural features, respectively. However, Yulin germplasm showed significantly higher proportions of cork and phloem, as well as stronger xylem vessel staining intensity compared to Daqing germplasm. Regarding various component contents, Yulin germplasm contained significantly higher levels of ethanol-soluble extracts, total saponins, and saikosaponins A, B2, C, D, while Daqing germplasm had significantly higher levels of hemicellulose, starch, and liposoluble extracts. After introduction to Yulin, the Daqing germplasm(YLDQC3) showed increased starch, water-soluble polysaccharides and liposoluble extracts contents, decreased cell wall component content, but no significant difference in other component contents. Metabolomics revealed that saponins and terpenes accumulated significantly in Yulin germplasm, while alcohols and aldehydes accumulated predominantly in Daqing germplasm. Transcriptomics indicated similar gene expression patterns within the same germplasm but specificity between different germplasms. Integrative metabolomic-transcriptomic analysis identified 145 potential key genes associated with the saikosaponin biosynthesis pathway, including one acetyl-coenzyme A(CoA) acetyltransferase gene(ACAT), one 3-hydroxy-3-methylglutaryl-coenzyme A synthase gene(HMGS), two hydroxymethylglutaryl-CoA(HMG-CoA) reductase genes(HMG), one phosphomevalonate kinase gene(PMK), one 1-deoxy-D-xylose-5-phosphate synthase gene(CLA), one hydroxymethylbuten-1-aldol synthase gene(HDR), two farnesyl pyrophosphate synthase genes(FPPS), one squalene synthase gene(SQS), one β-amyrin synthase gene(BAS), 102 cytochrome P450(CYP450) gene family members, and 32 uridine diphosphate-glucuronosyltransferase(UGT) gene family members. ConclusionAmong the three cultivated types, YLC3 most closely resembles YLW in appearance, microscopic features, contents of major bioactive constituents, metabolomic and transcriptomic profiles. Yulin germplasm exhibits superior saponin synthesis capability compared to Daqing germplasm, and Yulin region is more suitable for the growth of B. scorzonerifolium. Based on these findings, it is recommended that artificial cultivation in northern Shaanxi and similar regions utilize the local Yulin germplasm source cultivated for at least three years.
9.Comparison of Wild and Cultivated Polygalae Radix Based on Traditional Quality Evaluation
Yihan WANG ; Yanmeng LIU ; Huaizhu LI ; Yongliang LI ; Shan GUAN ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):215-224
ObjectiveBased on the traditional quality evaluation methods summarized in previous dynasties, this paper systematically contrasted the quality differences between wild Polygalae Radix(WPR) and cultivated Polygalae Radix(CPR) from the aspects of character, microscope and chemical composition by modern scientific and technological means, providing a basis for high-quality production and quality control. MethodsCPR and local WPR in Yulin city, Shaanxi province from 1 to 6 years were collected, and a systematic comparative analysis was conducted using traditional pharmacognosy research methods combined with modern multi-omics analysis techniques, including character traits(length, weight, diameter), cross-sectional microscopic features(proportions of cork, phloem, xylem, etc), cell wall component content(hemicellulose, cellulose, lignin), extracts content(water-soluble extract and alcohol-soluble extract), carbohydrate content(starch, water-soluble polysaccharides), contents of total flavonoids, total saponins and specific marker compounds(3,6′-disinapoyl sucrose, polygalaxanthone Ⅲ, tenuifoliside A, tenuifoliside C, sibiricose A5 and A6) and other indexes. Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was employed to conduct comparative analysis of secondary metabolites in WPR and CPR, and multivariate statistical analysis such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were combined to screen the key differential components of them. ResultsIn terms of appearance, there were significant differences between WPR and CPR. The characteristics of WPR conformed to the "thick wrinkles on the epidermis" recorded in ancient books, featuring a wrinkled surface and grayish-brown appearance. However, CPR had a finer texture and a yellowish white appearance, with weight, length, and diameter increasing with longer cultivation periods. In terms of microscopy, WPR exhibited a thick cork layer with fissures in the phloem, whereas CPR had a thinner cork layer with uniformly arranged cork cells. Younger PR specimens showed numerous phloem fissures in cross-sections, while older specimens display progressively denser arrangements of phloem parenchyma cells. In terms of the contents of various major components, the contents of water-soluble extract, starch and total saponins in WPR were inversely proportional to the root diameter, while the contents of water-soluble extract, water-soluble polysaccharides and total saponins in CPR decreased with the increase of planting years. The content of xanthones in WPR was significantly higher than that of CPR, while the contents of other major components showed no significant change pattern. Among the six indicator components, the average content of sibiricose A5 in WPR was significantly higher than that of CPR, followed by slightly higher content of tenuifoliside A. In CPR, the relative content of 3,6′-disinapoyl sucrose and tenuifoliside A was the highest. The former showed an increase in volatility with increasing cultivation years, while the latter showed a decrease in volatility. The results of differential compound analysis based on UPLC-Q-TOF-MS showed that there were significant differences in metabolites between WPR and CPR samples. Among them, the seven compounds with the largest differences among WPR samples of different thicknesses were polygalasaponins, and for CPR with different planting years, the main differential compounds were oligosaccharide esters. ConclusionThere are differences between WPR and CPR in character, microscopic structure and chemical composition, and some components are inversely proportional with the increase of diameter and cultivation duration due to the distribution characteristics. However, the longer the cultivation years of PR, the closer it is to the "thick wrinkles on the epidermis" of WPR, which has been respected by generations. It is suggested that this traditional character combined with modern component contents should be used as the index of artificial cultivation and quality control of PR.
10.Comparison of Wild and Cultivated Alpiniae Oxyphyllae Fructus Based on Traditional Quality Evaluation
Fengfan WANG ; Yajie XIANG ; Jian FENG ; Wencheng HOU ; Wenlan LI ; Yangyang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):235-244
ObjectiveTo compare the differences between wild Alpiniae Oxyphyllae Fructus(WAOF) and cultivated Alpiniae Oxyphyllae Fructus(CAOF) through a traditional quality evaluation system for medicinal materials. MethodsA total of 10 batches of WAOF and 12 batches of CAOF samples were collected from various regions of Hainan province. Relevant analytical methods from the 2020 edition of the Pharmacopoeia of the People's Republic of China were employed to observe the characteristics of WAOF and CAOF, followed by microscopic identification, thin-layer chromatography(TLC) identification, moisture content(toluene method), total ash, acid-insoluble ash, water-soluble and alcohol-soluble extracts(hot dipping method), water-soluble protein, total polysaccharides and total flavonoids(ultraviolet spectrophotometry), and volatile oil content(method A under general rule 2204). The contents of five active components(protocatechuic acid, chrysin, kaempferol, tectochrysin and nootkatone) were quantified using ultra-performance liquid chromatography(UPLC), and the antioxidant activity was evaluated. Building upon traditional quality evaluation of AOF, quantitative measurements were conducted on its appearance traits including diameter, length, plumpness(diameter/length ratio), and color. Canonical correlation analysis was performed using SPSS 26.0 to explore relationships between appearance traits and intrinsic quality. ResultsNo significant differences were observed between WAOF and CAOF in microscopic observation, TLC identification, moisture content, protocatechuic acid content, kaempferol content, odor, or antioxidant activity measured by 2,2ʹ-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) method. WAOF exhibited significantly higher levels in water-soluble extracts, alcohol-soluble extracts, total polysaccharide content, water-soluble protein content, 100-grain weight, length, and total color difference(ΔE*ab) compared to CAOF(P<0.01). In contrast, CAOF showed significantly higher levels of total ash, acid-insoluble ash, content of total flavonoids, volatile oil content, chrysin content, tectochrysin content, nootkatone content, diameter, plumpness, lightness(L*), red-green chromaticity(a*), yellow-blue chromaticity(b*), and antioxidant activity measured by 1,1-diphenyl-2-picrylhydrazyl(DPPH) method compared to WAOF(P<0.01). Correlation analysis between 7 phenotypic traits and 8 quality traits revealed that among the phenotypic traits, plumpness, L*, a*, and b* exerted significant influence on intrinsic quality. Among the quality traits, total flavonoids, volatile oils, nootkatone, chrysin, and tectochrysin contributed substantially to intrinsic quality. ConclusionPlumpness, L*, a*, and b* of AOF significantly influence its intrinsic quality, and higher values of these parameters indicate relatively superior intrinsic quality. The comprehensive quality evaluation reveals that CAOF samples collected in this study are superior to their wild counterparts.


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