1.Optimization of Extraction Process and Material Basis Analysis of Modified Banxia Xiexintang
Yuling LIU ; Ruying TANG ; Dongxue ZHENG ; Qiling ZHANG ; Xinmin LIU ; Donghan BAI ; Longfei LIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):129-139
ObjectiveTo optimize and validate the optimal sequential alcohol-water extraction process of modified Banxia Xiexintang(MBXT) based on pharmacodynamic evaluation, combined with the G1-entropy weight method and Box-Behnken response surface methodology, and to systematically and comprehensively analyze the material basis of this formula, providing a scientific basis for its quality control and industrial production. MethodsRats were randomly divided into the normal group, model group, metformin group, and MBXT water extraction, water extraction and ethanol precipitation, sequential ethanol-water extraction groups. Except for the normal group, a polycystic ovary syndrome with insulin resistance(PCOS-IR) model was established in all rats via a high-fat diet combined with letrozole induction. Enzyme-linked immunosorbent assay(ELISA) biochemical assay kits and hematoxylin-eosin(HE) staining were used to compare sex hormone levels in serum and ovarian histopathology, thereby screening extraction process routes. Based on this, a comprehensive score was constructed using the G1-entropy weight method based on the transfer rates of index components(berberine hydrochloride and baicalin) and the dry extract rate. Box-Behnken response surface methodology was then utilized to optimize the extraction process parameters. Finally, the chemical constituents of the sample from the optimal process were qualitatively analyzed by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap mass spectrometry(UPLC-Q-Orbitrap-MS). ResultsPharmacodynamic findings revealed that compared with the normal group, serum testosterone(T) and luteinizing hormone(LH) levels were significantly elevated in the model group, while estradiol(E2) and follicle-stimulating hormone(FSH) levels were significantly decreased(P<0.01), with polycystic changes observed in ovarian tissues. Compared with the model group, all treatment groups significantly reversed the changes in sex hormone levels, with the sequential ethanol-water extraction group showing the optimal effect in improving the aforementioned indicators and pathological morphology, followed by subsequent process optimization. The optimized process involved adding 12 times the amount of 70% ethanol for extracting twice, each lasting 120 min, and adding 12 times the amount of water for extracting thrice, each lasting 90 min. Validation test results showed that under optimal process conditions, the average transfer rates of berberine hydrochloride and baicalin were 76.05% and 93.38%, respectively. MS analysis identified a total of 377 compounds, including 112 flavonoids, 41 terpenoids, 28 organic acids, 22 coumarins, and 8 alkaloids, while elucidating the cleavage patterns of key components. ConclusionThe optimized sequential ethanol-water extraction process is stable and feasible, effectively preserving the material basis of MBXT for treating PCOS-IR. It further clarifies the main chemical composition of this formula, providing a scientific basis for the development and quality control of its preparations.
2.Exploring Mechanism of Modified Banxia Xiexintang in Ameliorating Metabolic Disorders and Reproductive Function in PCOS-IR Rats Based on Metabolomics and Transcriptomics
Donghan BAI ; Ruying TANG ; Longfei LIN ; Yuling LIU ; Dongxue ZHENG ; Qiling ZHANG ; Xinmin LIU ; Hui LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):140-149
ObjectiveTo evaluate the therapeutic effects of modified Banxia Xiexintang(MBXT) on polycystic ovary syndrome with insulin resistance(PCOS-IR) rats and reveal its potential mechanisms based on the integrated analysis of transcriptomics and metabolomics. MethodsFemale SD rats were selected, and a PCOS-IR model was established by intragastric administration of letrozole combined with a high-fat diet for 21 days. The modeled rats were randomly divided into the model group, MBXT low-, medium-, and high-dose groups(6.62, 13.23, 26.46 g·kg-1), and metformin group(0.158 g·kg-1), with a normal group set up separately. After 14 days of administration, the estrous cycle was observed, ovarian morphology was examined by hematoxylin-eosin(HE) staining, and the levels of testosterone(T), estradiol(E2), follicle-stimulating hormone(FSH), and luteinizing hormone(LH) in serum were detected by enzyme-linked immunosorbent assay(ELISA). Serum metabolites and ovarian tissue gene expression were detected using ultra-performance liquid chromatography-quadrupole-electrostatic orbitrap mass spectrometry(UPLC-Q-Orbitrap-MS) and RNA-Seq technology, respectively, followed by multi-omics integrated analysis. ResultsPharmacodynamic findings revealed that all MBXT dose groups could reversed abnormal estrous cycles in PCOS-IR rats, improve polycystic ovarian lesions, and normalize dysregulated serum hormone levels(T, LH, E2, FS, P<0.05, P<0.01). Metabolomic analysis revealed that compared with the model group, MBXT reversed 278 differential metabolites such as estrone and S-formylglutathione, mainly involving pathways such as steroid hormone biosynthesis, glutathione metabolism, and lipid peroxidation regulation. Transcriptomic analysis identified 434 differentially expressed genes, and enrichment analysis revealed that MBXT significantly regulated lipid peroxidation defense systems, including glutathione metabolism, peroxisome function, and fatty acid metabolism, thereby intervening in ferroptosis processes. It also engaged in inflammation-related pathways such as the chemokine signaling pathway. Integrated analysis revealed that both metabolomics and transcriptomics co-enriched metabolic pathways associated with ferroptosis and fatty acid metabolism. And key Hub genes[such as Ras-related C3 botulinum toxin substrate 2 gene(Rac2) and Fas ligand gene(Faslg)] showed significant correlations with differential metabolites. ConclusionMBXT can effectively ameliorate reproductive dysfunction and metabolic disorders in PCOS-IR rats. Its mechanism may be related to remodeling the immune-metabolism network, particularly by regulating MHC-mediated immune responses, inhibiting local ovarian ferroptosis, and enhancing steroid hormone synthesis pathways.
3.Investigation on Mechanism of Modified Banxia Xiexintang in Improving Ovarian Dysfunction of PCOS-IR Rats by Inhibiting Ferroptosis via AMPK/FASN/GPX4 Signaling Pathway
Donghan BAI ; Ruying TANG ; Longfei LIN ; Yuling LIU ; Dongxue ZHENG ; Qiling ZHANG ; Xinmin LIU ; Hui LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):150-160
ObjectiveTo investigate the mechanism of modified Banxia Xiexintang(MBXT) in improving ovarian dysfunction in polycystic ovary syndrome with insulin resistance(PCOS-IR) rats by inhibiting ferroptosis through the adenosine monophosphate(AMP)-activated protein kinase(AMPK)/fatty acid synthase(FASN)/glutathione peroxidase 4(GPX4) signaling pathway. MethodsSeventy-six female SD rats were randomly divided into a normal group(n=13) and a modeling group(n=63). The modeling group established a PCOS-IR model by intragastric administration of letrozole combined with a high-fat diet for 21 days. After successful modeling, these rats were randomly divided into the model group, MBXT low-, medium-, and high-dose groups(6.62, 13.23, 26.46 g·kg-1), metformin group(0.158 g·kg-1), and high-dose of MBXT combined with ferroptosis inducer Erastin group(15 mg·kg-1), with 10 rats in each group. After 14 days of intervention, ovarian pathological morphology was observed by hematoxylin-eosin(HE) staining, the mitochondrial ultrastructure of granulosa cells was observed by transmission electron microscopy(TEM), ovarian reactive oxygen species(ROS) levels were detected by dihydroethidium(DHE) probe, biochemical methods were used to detect Fe2+, malondialdehyde(MDA), glutathione(GSH) and other indicators in ovarian tissues, serum sex hormone and insulin levels were measured by enzyme-linked immunosorbent assay(ELISA), and the protein expressions of AMPK, FASN, acyl-CoA synthetase long-chain family member 4(ACSL4), GPX4, and solute carrier family 7 member 11(SLC7A11) in ovarian tissues were detected by Western blot. ResultsCompared with the normal group, the model group showed polycystic changes in the ovaries, with atrophy of mitochondria in granulosa cells and increased membrane density. Serum levels of testosterone(T), luteinizing hormone(LH), and insulin were significantly increased(P<0.01). The levels of ROS, MDA, 4-hydroxynonenal(4-HNE), and Fe2+ in ovarian tissues were significantly elevated(P<0.01), while adenosine triphosphate(ATP), GSH, and reduced nicotinamide adenine dinucleotide phosphate (NADPH) levels were significantly decreased(P<0.01). The phosphorylation levels of AMPK and acetyl-CoA carboxylase (ACC), as well as the protein expressions of SLC7A11, GPX4, and ferroptosis suppressor protein 1(FSP1) were significantly downregulated(P<0.01), whereas the expressions of FASN, ACSL4, and nuclear receptor coactivator 4(NCOA4) were significantly upregulated(P<0.01). Compared with the model group, MBXT intervention at various doses improved the above pathological changes and biochemical indicators in a dose-dependent manner, with the high-dose group showing the most significant effect(P<0.01). Compared with the MBXT high-dose group, the high-dose of MBXT combined with ferroptosis inducer Erastin group restored ovarian ferroptosis characteristics in rats, with increased ROS and lipid peroxidation products, and altered expressions of key proteins(P<0.05, P<0.01). ConclusionMBXT can effectively improve ovarian function and metabolic disorders in PCOS-IR rats. Its mechanism may be related to activating the AMPK/ACC signaling pathway, downregulating FASN and ACSL4 to reduce lipid peroxidation substrates, and restoring glucose-6-phosphate dehydrogenase/phosphoglycerate dehydrogenase(G6PD/PHGDH) metabolic flux to enhance the GPX4/FSP1 antioxidant defense system, thereby inhibiting ferroptosis in ovarian granulosa cells.
4.Integrative Strategies of Traditional Chinese and Western Medicine for Polycystic Ovary Syndrome with Insulin Resistance: Synergistic Potentiation Mechanisms Based on Pathogenic Complementarity
Ruying TANG ; Hengchang LIU ; Longfei LIN ; Donghan BAI ; Yuling LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):161-171
Polycystic ovary syndrome (PCOS) with insulin resistance (IR) represents a common complex endocrine-metabolic disorder in reproductive-aged women,posing substantial threats to their reproductive and long-term health. The networked characteristics of pathological mechanisms pose severe challenges in the diagnosis and treatment modes with a single medical system. Western medicine identifies it as a pathological axis with the core of "IR-hyperinsulinemia-hyperandrogenemia" and offers effective symptomatic treatments,it is often faced with limitations such as insufficient overall regulation and drug side effects. Traditional Chinese medicine is characterized by a holistic concept centered on the dysfunction of "kidney-spleen-liver" and syndrome differentiation and treatment,yet its microscopic action mechanisms remain to be fully elucidated. An integrative diagnosis and treatment mode of traditional Chinese medicine and western medicine provides a vital approach to overcoming these challenges. This review systematically sorted out the understanding of polycystic ovary syndrome with insulin resistance (PCOS-IR) pathogenesis from both Chinese and western medicine perspectives. An explainable mapping relation potentially existing between the molecular pathological networks defined by western medicine and the macroscopic dialectical system of traditional Chinese medicine was hypothesized. The synergistic potentiation mechanisms of integrative strategies of traditional Chinese medicine and western medicine were analyzed,with a focus on clarifying the recent progress of integrative diagnosis and treatment strategies. The paper aims to provide a more comprehensive and profound reference for research and clinical practice in this field,advance the further development of the prevention and treatment of PCOS-IR with integrated traditional Chinese medicine and western medicine,and explore the establishment of a more precise,individualized diagnosis and treatment system to make optimal clinical decisions.
5.Ameliorative effect and mechanism of vitexin on inflammation in ulcerative colitis mice
Lin ZHOU ; Pengfei XIA ; Yuling LIU ; Zhichao MENG ; Geng LI ; Yuanyuan YU
China Pharmacy 2026;37(6):758-763
OBJECTIVE To explore the ameliorative effect and potential mechanism of vitexin on inflammation in ulcerative colitis (UC) mice. METHODS The UC mice model was established by continuous administration of 3% dextran sulfate sodium solution for 5 days. Mice with successful modeling were randomly divided into UC group, vitexin low- and high-dose groups (vitexin-L and vitexin-H groups, 40, 80 mg/kg), mesalazine group (400 mg/kg), and vitexin-H+recombinant Jagged canonical Notch ligand 1 (rJagged-1) group (vitexin-H+rJagged-1 group, 80 mg/kg vitexin+1 mg/kg rJagged-1), with 12 mice in each group. Another 12 normal mice were used as the control (CK) group. Mice in each group were administered the corresponding drugs or the corresponding drugs and normal saline by gavage and intraperitoneal injection once daily for 7 consecutive days. General conditions were observed during the experiment. At 24 h after the last administration, the disease activity index (DAI) score was evaluated. Colonic histopathological morphology was observed and scored. Macrophage polarization levels in the spleen and colon tissues were measured. The protein expressions of interleukin-6 (IL-6), IL-10, tumor necrosis factor-α (TNF-α), transforming growth factor-β 1 (TGF-β 1 ), Jagged-1, Notch1 and Notch intracellular domain (NICD) in colonic tissues were determined. RESULTS Compared with the UC group, the symptoms (reduced food and water intake, dull fur, etc.) and pathological changes (epithelial cell shedding, inflammatory cell infiltration, etc.) were significantly improved in the vitexin-L, vitexin-H and mesalazine groups. DAI scores, colonic histopathological scores, M1 macrophage contents in spleen tissue, M1/M2 macrophage ratios, M1 macrophage proportions in colon tissue, and protein expressions of IL-6, TNF-α, Jagged-1, Notch1 and NICD in colon tissue were significantly decreased ( P <0.05). Meanwhile, the M2 macrophage contents in spleen tissue, M2 macrophage proportions in colon tissue, and protein expressions of IL-10 and TGF-β 1 in colon tissue were significantly increased ( P <0.05). Moreover, the improvement effects in the vitexin-H and mesalazine groups were significantly superior to those in the vitexin-L group ( P <0.05). Compared with the vitexin-H group, the above symptoms and pathological changes were aggravated, and all quantitative indicators were significantly reversed in the vitexin-H+rJagged-1 group ( P <0.05). CONCLUSIONS Vitexin can ameliorate the inflammation of UC mice, which is associated with its inhibition of the Jagged-1/Notch1 pathway and regulation of macrophage polarization (inhibition of M1-type polarization and promotion of M2-type polarization).
6.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.
7.Effect of ankle soft robotic exosuit on gait function for stroke patients: a scoping review from 2020 to 2025
Huigang GAO ; Yuanwen LIU ; Yuling QIN ; Xiquan HU ; Lin ZHU
Chinese Journal of Rehabilitation Theory and Practice 2026;32(6):645-652
ObjectiveTo explore effect of the ankle soft robotic exosuits (ASRE) on gait function in stroke patients. MethodsDatabases including PubMed, Embase, Scopus, Web of Science, CNKI, Wanfang and VIP were searched for researches evaluating the impact of ASRE on gait function in stroke patients with hemiplegia, covering the period from January 1st, 2020 to May 1st, 2025. The PEDro scale and the Risk of Bias in Non-randomized Studies of Interventions tool were used to assess the quality of the included researches, and data were extracted for a scoping review. ResultsA total of twelve researches were finally included, consisting of three randomized controlled trials (RCT), three cross-over controlled trials and six pre-post controlled trials, which came from three countries of China, the United States and South Korea, involving 256 patients. The three RCT were rated as high-quality, while the remaining researches were presented moderate to serious risk of bias. Five researches employed a single-session intervention; and seven used multiple-session intervention, with 30 to 60 minutes a session, two to five sessions a week, for five to 20 sessions. ASRE was effective on walking speed, walking distance, propulsive force of the hemiplegic side, gait parameters and spatiotemporal symmetry, joint range of motion, as well as the scores of scales related to lower limb motor function, balance and gait, with favorable safety and reliability. ConclusionASRE may improve the gait function in stroke patients.
8.The effectiveness of the peripheral arterial calcification scoring system based on CT angiography in assessing renal function in patients with peripheral arterial disease
Yuling CUI ; Ningning DING ; Li ZHOU ; Yan MENG ; Yaqing HAN ; Cuilin YIN ; Zhe LIU ; Jian YANG
Journal of Practical Radiology 2025;41(4):589-593
Objective To explore the effectiveness of the peripheral arterial calcification scoring system(PACSS)based on computed tomography angiography(CTA)in assessing renal function in patients with peripheral arterial disease(PAD).Methods The clinical data,CTA imaging data,and laboratory results from PAD patients who underwent lower limb artery CTA examination were retrospectively collected.The PACSS was utilized to score the calcification in both lower limb arteries.Participants were categorized into three groups based on their estimated glomerular filtration rate(eGFR)(normal group:eGFR≥90 mL/min;mild renal dysfunction group:eGFR 60-89 mL/min,and moderate to severe renal dysfunction group:eGFR<60 mL/min).The demographic data,clinical symptoms,and comorbidities among the three groups were compared by analysis of variance(ANOVA).The Spearman correlation coefficient was employed to evaluate the relationship between eGFR,cystatin C,and PACSS score.Results The age(P<0.001)and PACSS score(P<0.05)of patients with renal dysfunction were significantly higher than those of patients with normal renal function.However,there were no significant differences in gender,prevalence of diabetes,hypertension,or severe limb ischemia.Spearman correlation analysis showed that eGFR was negatively correlated with PACSS score(r=-0.18 in the right lower limb,P=0.037,r=-0.24 in the left lower limb,P=0.006).In contrast,cystatin C was positively correlated with PACSS score(r=0.26 in the right lower limb,P<0.001,r=0.22 in the left lower limb,P=0.002).Conclusion The PACSS score of lower limb artery in PAD patients is corre-lated with the severity of renal dysfunction.This finding may facilitate early warning and clinical intervention for PAD patients with renal dysfunction.
9.Management Suggestions and Catalog Recommendations for Emergency Medicines After Earthquake in High-altitude Areas
Yuling BAO ; Xia GUO ; Yu LIU ; Dong LIU
Herald of Medicine 2025;44(3):349-355
Based on the special environment of high-altitude areas and the characteristics of post-disaster medicines de-mand,this article discussed the difficulties and management strategies for post-earthquake emergency medicines support.Natural conditions such as low temperature,dryness and ultraviolet radiation in high-altitude areas,as well as problems such as weak transportation infrastructure and shortage of medical resources,have posed huge challenges to the stability,transportation,supply,distribution and use of the medicines.Therefore,this paper suggested solutions such as diversified supply channels,transportation reinforcement,temperature and humidity control,light protection measures,sand and corrosion prevention,proposed precautions such as medicine distribution on demand,rational use and strengthening of pharmaceutical care,also emphasized that local cultur-al habits should be respected.Furthermore,according to high-altitude areas environmental characteristics and different rescue sta-ges(emergency period,sub-emergency period,recovery period),the corresponding emergency medicines catalog was recommen-ded.The study aims to provide scientific basis and practical guidance for the medicine supply of earthquake in plateau,improve rescue efficiency,and reduce disaster losses.
10.De Novo Assembly and Phylogenetic Study of Chloroplast Genomes of Five Species of Genus Polygonatum
Wei LI ; Mingyu ZHU ; Yuling ZENG ; Xuan WEN ; Chutong HUANG ; Xinyue FA ; Lin SEN ; Zhigang HU ; Yifei LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(1):36-55
Objective Five chloroplast(cp)genomes from members of genus Polygonatum were assembled by hybrid assembly technique,and their intraspecic and interspecific differences were analyzed by comparative genomic method.Codon usage patterns and influencing factors were determined,and the cp genome data were applied to understand the phylogenomic relationships in the entire genus Polygonatum along with the available data.Methods In this study,the chloroplast genomes of 5 species of genus Polygonatum were assembled using Unicycler software.Sequence alignment,collinearity analysis,boundary analysis and other methods were used to evaluate the interspecific differences of these five species.Nucleotide polymorphism analysis was used to discover the high-variation sites of the five species and their related species,predict the distribution of different long repeat sequences and SSRs,and then analyze the use bias of Polygonatum code.Finally,phylogenetic tree was constructed with the coding sequences of other 47 genus Polygonatum and their closely related species to explore their phylogenetic relationships in this study.Results ①Chloroplast genomes of 155 408-155 623 bp were assembled from five species of Polygonatum.A total of 132-133 genes were annotated,and 369 long repeats and 1553 simple repeats were detected.②The contraction and expansion of chloroplast genomes in 8 species were not obvious at the IRs boundary,and the size and distribution of individual genes at the LSC-IRs-SSC boundary,such as ndhF gene and ycf1 gene,were slightly different.No interspecific or intraspecific rearrangement was observed in 8 species.③ The high-variation regions of the 8 chloroplast genomes are mainly located in two single-copy regions,the duplicate copy region is relatively conserved,and the coding region is more conserved than the non-coding region.High nucleotide polymorphic loci rps16-trnQ,trnS-trnG,trnTUGU-trnL,ndhF-rpl32 and rpl32-trnL are located in the single copy region and most of them are gene spacer regions.④ The codon preference results showed that the codon preference of the five species was similar and mainly affected by selection pressure,and the third base of the codon played A dominant role and mainly ended in A/U.RSCU clustering heat map shows that PK and PZ,PCB and PS have close relationship.⑤ Phylogenetic trees divided 52 species into five branches:Ⅰ,Ⅱ,Ⅲ,ⅣandⅤ.PS,PK,PCB,PCZ and PZ were divided into ⅣandⅤbranches,among which PK and PZ were most closely related,while PCZ was more distant than the other four,was divided into the Ⅴbranch alone.Conclusion This study provided a reference for the phylogenetic research and molecular marker development of the medicinal plants of the Polygonum genus.

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