1.Pre-operative risk assessment of hepatocellular carcinoma recurrence in liver transplant recipients by non-invasive detection of pre-existing genetic lesions
Suqin YANG ; Sunbin LING ; Jianhua LI ; Yan WANG ; Jiapei WANG ; Qiwei HUANG ; Fanming LIU ; Yiqi ZHUANG ; Yingyu ZHENG ; Rui WANG ; Zhe YANG ; Xiaoping ZHENG ; Kai WANG ; Zhikun LIU ; Jun CHEN ; Jianguo WANG ; Haiyang XIE ; Lin ZHOU ; Leiming CHEN ; Guoqiang CAO ; Dandan CHEN ; Junfang JI ; Bin ZHAO ; Chao JIANG ; Di LU ; Xuyong WEI ; Hangjin JIANG ; Qiaonan SHAN ; Hengbo SHI ; Yong-Zhen XU ; Shusen ZHENG ; Zhengxin WANG ; Shengda LIN ; Xiao XU
Clinical and Molecular Hepatology 2026;32(2):884-903
Background/Aims:
Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy.
Methods:
We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues.
Results:
We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts.
Conclusion
Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.
2.Analysis of weight loss behavior and related factors of middle school students in Shanghai
CHEN Weili, ZHANG Zhe, ZHAI Yani, YAN Qiong, QI Yue, LUO Chunyan
Chinese Journal of School Health 2025;46(2):223-228
Objective:
To investigate the distribution characteristics and related factors of weight loss behavior among middle school students in Shanghai, so as to provide a reference for guiding scientific weight loss among middle school students.
Methods:
From May to June 2021, a stratified cluster random sampling method was used to select 16 758 junior and high school students in 16 districts of Shanghai. Youth Risk Behavior Surveillance System was administered to assess the basic condition and weight loss behaviors of the students. An unordered multinomial Logistic regression model was employed to analyze the factors associated with weight loss behaviors.
Results:
A total of 5 881 (35.09%) reported engaging in exercise for weight loss, 6 344 (37.86%) reported dieting for weight loss, and 461 (2.75%) engaged in unhealthy weight loss behaviors. The unordered multinomial Logistic regression analysis indicated that compared with the no weight loss behavior group, students from urban areas( OR =1.35,95% CI =1.10-1.66), those with Internet addiction ( OR =1.71,95% CI =1.23-2.38), those with victims of bullying ( OR =2.09, 95% CI =1.68-2.61), those experiencing insomnia ( OR =2.33,95% CI = 1.74-3.11), those feelings of sadness or despair ( OR =3.10, 95% CI =2.42- 3.97 ), and those who perceived their body weight as slightly heavy ( OR =2.77, 95% CI = 2.17-3.55) or very heavy ( OR =3.41, 95% CI =2.44-4.75) were more likely to engage in unhealthy weight loss behaviors ( P <0.05).
Conclusions
There are significant differences in weight loss behaviors among middle school students with varying characteristics in Shanghai. Negative emotions such as insomnia and feelings of sadness or despair, Internet addiction, cognitive bias in weight and experiences of bullying are identified as related factors for unhealthy weight loss behaviors. Targeted intervention measures should be implemented to guide students towards scientific approaches to weight management.
3.Ten surgical pearls adapted from ancient Chinese allusions in managing severe proliferative diabetic retinopathy
Zhe CHEN ; Chan WU ; Yan ZHOU ; Shiqun LIN ; Xingyu XIAO ; Rongping DAI
International Eye Science 2025;25(5):698-705
AIM: To summarize 10 surgical pearls for managing proliferative diabetic retinopathy(PDR)adapted from the ancient Chinese allusions and analyze the application of these pearls in a real-world fashion.METHODS: Retrospective, noncomparative, interventional study. Ten surgical pearls were summarized and adapted from the ancient Chinese philosophy. Totally 346 cases(443 eyes)that underwent pars plana vitrectomy(PPV)at our hospial from January 2016 to February 2024 were selected. Flexible combinations of these pearls were applied according to the specific condition of each patient during surgeries. The efficacy and safety were analyzed, as well as the application frequencies according to the existence of tractional retinal detachment or not.RESULTS: A total of 473 times of surgeries were performed on all the patients. According to ancient Chinese allusions, ten surgical pearls were summarized from these surgeries. All PPVs went smoothly with the application of different combinations. Finally, almost all proliferative membranes were successfully peeled except for 10 patients(11 eyes), who went through strategy No.10(minimal membranectomy)that, only necessary relaxation incisions were made with most of the proliferative membranes left on purpose. The final visual acuities were mostly improved or stable(1.92±0.83 LogMAR preoperatively vs 1.16±0.85 LogMAR postoperatively, P<0.01). Postoperative complications mainly included early inflammatory responses in the anterior chamber and nuclear sclerosis. Recurrent vitreous hemorrhage, retinal detachment, and hyphema or neovascular glaucoma occurred in 1.9%(9/473), 3.2%(15/473), 0.4%(2/473)and 0.4%(2/473)times of PPVs, respectively. After 12/473(2.5%)times of PPVs, retinal detachment at the macular area still existed, and multiple times of subsequent PPVs were conducted. Final retinal attachment at the macular area was realized in 98.9% eyes. Those 5 unattached eyes were with heavily reproliferated membranes and subsequent tractional retinal detachment recurrence under the oil, and three of them were scleral buckled additionally.CONCLUSION:These 10 surgical strategies and technique pearls were mostly effective and safe in the management of severe PDR patients. They were relatively easy to be memorized and applicated once the meaning of each Chinese idiom was understood. One can use different combinations flexibly according to a patient's specific condition.
4.Modulation of colonic DNA methyltransferase by mild moxibustion and electroacupuncture in ulcerative colitis TET2 knockout mice
Gege FENG ; Yue ZHANG ; Huangan WU ; Lu ZHU ; Hongxiao XU ; Zhe MA ; Yan HUANG
Digital Chinese Medicine 2025;8(1):100-110
Objective:
To investigate the mechanism of in alleviating colonic mucosal inflammation in ten-eleven translocation (TET) protein 2 gene knockout (TET2-/-) mice with ulcerative colitis (UC) by regulating DNA methyltransferase (DNMT) and DNA hydroxymethylase.
Methods:
Male specific pathogen-free (SPF) grade C57BL/6J wild-type (WT) mice (n = 8) and TET2-/- mice (n = 20) were used to establish UC models by freely drinking 3% dextran sulfate sodium solution for 7 d. After UC model validation through histopathological examination in two mice from each type, the remaining mice were divided into four groups (n = 6 in each group): WT model (WT + UC), TET2-/- model (TET2-/- + UC), TET2-/- mild moxibustion (TET2-/- + MM), and TET2-/- electroacupuncture (TET2-/- + EA) groups. TET2-/- + MM group received mild moxibustion on Tianshu (ST25) and Qihai (CV6) for 10 min daily for 7 d. The TET2-/- + EA group also applied electroacupuncture (1 mA, 2/100 Hz) at the same acupoints for 10 min daily for 7 d. The disease activity index (DAI) scores of each group of mice were accessed daily. The colon lengths of mice in groups were measured following intervention. The pathological changes in the colon tissues were observed with hematoxylin and eosin (HE) staining. The concentrations of interleukin (IL)-6, C-C motif chemokine 17 (CCL17), and C-X-C motif chemokine ligand 10 (CXCL10) in serum were detected by enzyme-linked immunosorbent assay (ELISA). The expression of DNMT proteins (DNMT1, DNMT3A, and DNMT3B) in the colon tissues was detected by immunohistochemistry. The expression of 5-methylcytosine (5-mC), 5-hydroxymethylcytosine (5-hmC), histone deacetylase 2 (HDAC2), and DNA hydroxymethylase family proteins (TET 1 and TET3) was detected using immunofluorescence, which also determined the co-localization of TET1 and IL-6 protein.
Results:
Compared with WT + UC group, TET2-/- + UC group exhibited significantly higher DAI scores and shorter colon lengths (P < 0.01). Both mild moxibustion and electroacupuncture significantly decreased DAI scores and ameliorated colon shortening in TET2-/- mice (P < 0.001). Histopathological scores of TET2-/- + UC mice were significantly higher than those of WT + UC group (P < 0.001) and were significantly reduced after both mild moxibustion and electroacupuncture interventions (P < 0.001). Serum levels of IL-6, CCL17, and CXCL10 were significantly elevated in TET2-/- + UC group compared with WT + UC group (P < 0.001). Mild moxibustion significantly reduced IL-6, CCL17, and CXCL10 levels (P < 0.001, P < 0.001, and P < 0.01, respectively), while electroacupuncture also significantly reduced IL-6, CCL17, and CXCL10 levels (P < 0.05, P < 0.01, and P < 0.01, respectively). TET2-/- + UC mice showed increased expression levels of DNMT1, DNMT3A , DNMT3B, and 5-mC (P < 0.05, P < 0.01 and P < 0.001, respectively), with decreased expression levels of TET1, TET3, 5-hmC, and HDAC2 (P < 0.001). Mild moxibustion significantly reduced DNMT1, DNMT3B, and 5-mC levels (P < 0.05, P < 0.01, and P < 0.001, respectively), while increasing expression levels of TET1, TET3, 5-hmC, and HDAC2 (P < 0.001, P < 0.001, P < 0.05, and P < 0.001, respectively). Electroacupuncture significantly decreased 5-mC and DNMT3B levels (P < 0.001 and P < 0.01, respectively) and increased 5-hmC and HDAC2 levels (P < 0.05 and P < 0.001, respectively), but did not significantly affect TET1 and TET3 expression (P > 0.05). Compared with TET2-/- + MM group, TET2-/- + EA group showed significantly higher 5-mC expression (P < 0.001). TET2-/- + UC group exhibited markedly increased IL-6 expression and higher co-localization of TET1 and IL-6 in mucosal epithelium, whereas minimal IL-6 expression was observed in the other groups.
Conclusion
Mild moxibustion and electroacupuncture significantly ameliorate colonic inflammation exacerbated by TET2 deficiency in UC mice via epigenetic modulation. Distinct mechanisms exist between the two interventions: mild moxibustion regulates both DNMT and hydroxymethylase, whereas electroacupuncture primarily affects DNMT.
5.Processing History and Modern Research of Jianghuanglian: A Review
Ying LI ; Yun WANG ; Zhe JIA ; Lin YAN ; Min JIN ; Cun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):275-282
Jianghuanglian is one of the representative processed products of Coptidis Rhizoma for treating cold syndrome with drugs of heat nature, and ginger is used to restrict the bitter cold of Coptidis Rhizoma, which can be traced back to Bojifang, and it is suitable for stagnation of damp-heat in middle-jiao, cold-heat mutual knots and other symptoms. Jianghuanglian retains the alkaloids, phenylpropanoids and flavonoids of Coptidis Rhizoma, and also introduces gingerol components such as 6-gingerol in ginger, which has pharmacological activities such as anti-inflammatory, antibacterial, anti-tumor, and improving gastrointestinal function. The 2020 edition of Chinese Pharmacopoeia and many local processing specifications have included the traditional processing process and quality standards of Jianghuanglian, but the specific process parameters and quality standards are incomplete, which limits the production and clinical application of this processed product. By summarizing the processing history, process research, quality evaluation, pharmacodynamic and medicinal property changes and application of Jianghuanglian in the past 20 years, there are differences in the processing methods and standards in various provinces and cities, which are mainly reflected in the preparation method, dosage, processing process and quantitative standards of ginger juice. In addition, there are also certain differences in the changes of the main components of Jianghuanglian prepared from ginger or dried ginger, as well as their efficacy and medicinal properties. The research on the processing process of Jianghuanglian plays an important role in improving its quality standards, and this review can provide a reference for improving the quality evaluation system of Jianghuanglian.
6.Efficacy and Safety of Yangxue Qingnao Pills Combined with Amlodipine in Treatment of Hypertensive Patients with Blood Deficiency and Gan-Yang Hyperactivity: A Multicenter, Randomized Controlled Trial.
Fan WANG ; Hai-Qing GAO ; Zhe LYU ; Xiao-Ming WANG ; Hui HAN ; Yong-Xia WANG ; Feng LU ; Bo DONG ; Jun PU ; Feng LIU ; Xiu-Guang ZU ; Hong-Bin LIU ; Li YANG ; Shao-Ying ZHANG ; Yong-Mei YAN ; Xiao-Li WANG ; Jin-Han CHEN ; Min LIU ; Yun-Mei YANG ; Xiao-Ying LI
Chinese journal of integrative medicine 2025;31(3):195-205
OBJECTIVE:
To evaluate the clinical efficacy and safety of Yangxue Qingnao Pills (YXQNP) combined with amlodipine in treating patients with grade 1 hypertension.
METHODS:
This is a multicenter, randomized, double-blind, and placebo-controlled study. Adult patients with grade 1 hypertension of blood deficiency and Gan (Liver)-yang hyperactivity syndrome were randomly divided into the treatment or the control groups at a 1:1 ratio. The treatment group received YXQNP and amlodipine besylate, while the control group received YXQNP's placebo and amlodipine besylate. The treatment duration lasted for 180 days. Outcomes assessed included changes in blood pressure, Chinese medicine (CM) syndrome scores, symptoms and target organ functions before and after treatment in both groups. Additionally, adverse events, such as nausea, vomiting, rash, itching, and diarrhea, were recorded in both groups.
RESULTS:
A total of 662 subjects were enrolled, of whom 608 (91.8%) completed the trial (306 in the treatment and 302 in the control groups). After 180 days of treatment, the standard deviations and coefficients of variation of systolic and diastolic blood pressure levels were lower in the treatment group compared with the control group. The improvement rates of dizziness, headache, insomnia, and waist soreness were significantly higher in the treatment group compared with the control group (P<0.05). After 30 days of treatment, the overall therapeutic effects on CM clinical syndromes were significantly increased in the treatment group as compared with the control group (P<0.05). After 180 days of treatment, brachial-ankle pulse wave velocity, ankle brachial index and albumin-to-creatinine ratio were improved in both groups, with no statistically significant differences (P>0.05). No serious treatment-related adverse events occurred during the study period.
CONCLUSIONS
Combination therapy of YXQNP with amlodipine significantly improved symptoms such as dizziness and headache, reduced blood pressure variability, and showed a trend toward lowering urinary microalbumin in hypertensive patients. These findings suggest that this regimen has good clinical efficacy and safety. (Registration No. ChiCTR1900022470).
Humans
;
Amlodipine/adverse effects*
;
Drugs, Chinese Herbal/adverse effects*
;
Male
;
Female
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Hypertension/complications*
;
Middle Aged
;
Treatment Outcome
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Drug Therapy, Combination
;
Adult
;
Blood Pressure/drug effects*
;
Double-Blind Method
;
Aged
;
Antihypertensive Agents/adverse effects*
7.Design and verification of accurate measurement of human body mass in microgravity environment
Zhe ZHANG ; Weibo LIU ; Zhi XU ; Yan ZHANG ; Jianping GUO ; Yu ZHANG ; Sheng Yuan WANG ; Yong XUAN ; Yue GAO ; Mi JIANG
Space Medicine & Medical Engineering 2025;36(1):50-57
Traditional mass measurement methods are not applicable in microgravity environments,and the main challenge for in-orbit body mass measurement technology based on inertial principles is to address the random errors brought about by the weightless environment.These include additional torques due to shifts in the center of mass,nonlinear accelerations due to non-rigid human bodies,mechanical energy consumption due to organ vibrations,and random vibrations of the measurement device itself.To address the above difficulties,the project proposes a technical scheme based on the principle of linear acceleration,designs and constructs a ground-specific air-floating experimental and simulation platform,studies key data such as motion trajectory,acceleration change,and vibration frequency amplitude during the mass measurement process,and simulates the changes in the center of mass and random vibrations of the human body in a weightless environment.The project has designed an adjustable posture bracket to adapt to changes in the center of mass,enhance body restraint,and greatly reduce shaking;it has also developed an integrated four-bar linkage motion guidance mechanism,high-precision integrated photoelectric distance measurement,and modular motion constant force measurement device to ensure the accurate measurement of acceleration and constant force data.The product has undergone simulation calculations,ground human applicability tests,and in-orbit applicability verification in the space station.Ground test results show that the device achieves a body mass measurement accuracy better than 0.5%,and the dispersion is better than 0.38%;after flight mission verification and evaluation,the in-orbit body mass measurement dispersion is less than 0.4%,which is superior to the SLAMMD,a mass measurement device of the same principle on the International Space Station,and is at the forefront internationally,achieving accurate body mass measurement.
8.Analysis of Ancient and Modern Literature of Xiaoyaosan and Examination of Its Key Information
Zhe WANG ; Jialei CAO ; Lyuyuan LIANG ; Yiping WANG ; Chen CHEN ; Weixiao LI ; Bingqi WEI ; Yinli LI ; Yongbin YAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):227-237
First recorded in an official medical book from the Northern Song Dynasty called Taiping Huimin Heji Ju Fang (Prescriptions of the Bureau of Taiping People's Welfare Pharmacy), Xiaoyaosan has been developed and refined over generations and is preserved to this day. It specializes in soothing the liver,resolving stagnation,fortifying the spleen,and nourishing blood. In this study,ancient traditional Chinese medicine (TCM) books and contemporary studies were reviewed to obtain information on Xiaoyaosan using bibliometrics,including its historical development,dosage,origin,processing methods,decoction dosage,and ancient and modern indications. Furthermore,a question regarding the presence of Zingiberis Rhizoma Recens and Menthae Haplocalycis Herba in Xiaoyaosan was investigated,and a table of key information on Xiaoyaosan was compiled,providing references for developing Xiaoyaosan preparations. According to the weight and measurement system of the Song dynasty,the contemporary equivalent formulation of the decocted Xiaoyaosan consists of 20.65 g of Glycyrrhizae Radix et Rhizoma and 41.3 g of Angelica Sinensis Radix,Poria,Paeoniae Radix Alba,Atractylodis Macrocephalae Rhizoma,and Bupleuri Radix. The formulation is processed to obtain a mixed powder with a particle size of 10 mesh. For each dose,8.25 g of the mixed powder is combined with 1 g of unprocessed Zingiberis Rhizoma Recens and 0.62 g of Menthae Haplocalycis Herba in 300 mL of water. The mixture is decocted until the volume reaches 210 mL,and the residue is then removed,with no specific timing required for administration. After the processing,each dose consists of approximately 0.75 g of Glycyrrhizae Radix et Rhizoma and 1.50 g of Radix Angelica Sinensis,Poria,Paeoniae Radix Alba,Atractylodis Macrocephalae Rhizoma,and Bupleuri Radix. Ancient medical literature shows that Xiaoyaosan primarily treats blood deficiency and overstrain,specifically for symptoms including heat caused by blood deficiency and fatigue,irregular menstruation,headache,eye soreness,pain in the ribs and limbs,and emaciation and bone steaming. In the Qing Dynasty,ZHANG Lu clearly proposed the pathogenesis of liver depression,and since then,the use of Xiaoyaosan in treating various syndromes associated with liver depression has been highly praised by physicians in the Qing dynasty and modern times. Xiaoyaosan has a wide application in modern clinical practices,involving digestive diseases,gynecological diseases,psychological diseases,nervous system diseases,and otorhinolaryngologic diseases. Moreover,it is most commonly used to treat depression and other diseases complicated with depression,hyperplasia of the mammary gland,etc. The key information on Xiaoyaosan and its clinical applications in ancient and modern times investigated in the study could serve as a scientific reference for in-depth research and extended clinical applications of the prescription.
9.Predictive value of albumin,hemoglobin,and multifactorial model for poor postoperative prognosis in elderly patients with meningiomas
Yan-Yu GONG ; Hong QU ; Si-Zhe FENG ; Chun-Yong YU ; Jin-Wei DU ; Jin JIANG
Medical Journal of Chinese People's Liberation Army 2025;50(4):418-426
Objective To explore the predictive value of albumin,hemoglobin and multifactorial model for poor postoperative prognosis in elderly patients with meningioma.Methods A retrospective analysis was conducted on 253 elderly patients who underwent meningioma surgery and were transferred to the neurosurgical intensive care unit(NICU)at General Hospital of Northern Theater Command from January 2019 to September 2021,serving as the modeling cohort.Another 227 elderly patients who were treated in NICU after meningioma surgery from November 2021 to June 2023 were used as the validation cohort.Patients in the modeling cohort were categorized into good prognosis group[Glasgow Coma Scale(GCS)score>7,n=161]and poor prognosis group(GCS≤7,n=92)based on the GCS.Univariate and multifactorial logistic regression analyses were performed on the modeling cohort to identify independent risk factors,and a multifactorial model for predicting poor postoperative prognosis in elderly patients with meningioma was constructed based on these factors.The predictive efficacy and accuracy of the model were evaluated using the area under the receiver operating characteristic(ROC)curve(AUC),sensitivity,specificity,Hosmer-Lemeshow goodness-of-fit test,and calibration curves.The predictive value of postoperative albumin,hemoglobin,and the multifactorial models for postoperative prognosis in elderly meningioma patients was assessed using restricted cubic spline modeling(RCS),decision curves(DCA),and validated using an external validation cohort to assess the stability of the model.Results Meningioma WHO grade Ⅱand Ⅲ(OR=3.994,95%CI 1.963-8.126),postoperative hypoalbuminemia(OR=2.194,95%CI 1.079-4.462),and postoperative anemia(OR=2.117,95%CI 1.096-4.089)were identified as independent risk factors for poor postoperative prognosis in elderly meningioma patients(P<0.05),while the use of analgesic/sedative medications was a protective factor(OR=0.388,95%CI 0.201-0.748,P<0.05).The Hosmer-Lemeshow test indicated that the constructed multifactorial model had a good fit accuracy(P=0.161).The AUC for predicting poor postoperative prognosis in elderly meningioma patients for postoperative albumin and hemoglobin were 0.545(95%CI 0.472-0.617)and 0.632(95%CI 0.561-0.702),respectively,and showed a nonlinear dose-response relationship with prognosis(P<0.01).DCA analysis results showed that the net benefit rate of multifactorial model was higher than that of postoperative albumin and hemoglobin when the threshold probabilities were between 0.10 and 0.90.The AUC for predicting postoperative prognosis in the elderly meningioma patients in the modeling and validation cohorts were 0.810 and 0.819,respectively,and their calibration curves suggested good discrimination and accuracy.Conclusions Meningioma WHO grades Ⅱ and Ⅲ,postoperative anemia and hypoalbuminemia are independent risk factors for poor postoperative prognosis in elderly meningioma patients,while the use of analgesic/sedative drugs is a protective factor.The multifactorial model constructed based on these factors has a good predictive efficacy and credibility,and can be used as a reference for clinical decision-making.
10.Microbial community mediated by microbial agents improves the quality of Epimedium pubescens Maxim.
Kunyang LAI ; Xiufu WAN ; Jiancai XIAO ; Hongyang WANG ; Shangxuan SHI ; Binbin YAN ; Chaogeng LYU ; Chengcai ZHANG ; Yufei ZHANG ; Feng YUAN ; Zhe ZHAO ; Shoudong ZHU ; Chuanzhi KANG ; Yan ZHANG
Science of Traditional Chinese Medicine 2025;3(3):270-281
Background: Optimizing cultivation techniques for traditional Chinese medicine has become a crucial means to improve the quality of medicinal materials. Microbial agents, as environmentally friendly and efficient plant growth promoters and soil conditioners, have increasingly attracted attention in eco-agriculture research. Objective: Our understanding remains limited regarding how the application of microbial agents, alone or in combination, affects changes in the rhizosphere microbiome and its association with the bioactive components of medicinal materials. Methods: In this study, Epimedium pubescens Maxim. was employed as a model plant to examine the effects of 2 microbial agents(Paenibacillus mucilaginosus and Bacillus subtilis) applied individually and in combination on plant growth and the accumulation of bioactive components. Additionally, this study explored the relationship between the rhizosphere microbiome and plant development. Results: The application of microbial agents increased the yield of E. pubescens leaves by 20.30% to 33.66% and enhanced the total flavonol glycosides content by 11.40% to 29.94%. Meanwhile, microbial treatments reshaped the rhizosphere microbiome, promoted the enrichment of beneficial microorganisms (e.g., Frankia and Paenibacillus), suppressed phytopathogenic fungi such as Didymella and Scytalidium, and enhanced the stability of the soil microbial co-occurrence network. The partial least squares path model suggested that microbial agents not only directly impact the quality of medicinal herbs but also indirectly alter the accumulation of bioactive components by modulating the soil microbiome. Conclusion: These findings deepen our understanding of the relationship between medicinal plant quality and rhizosphere microbiomes as mediated by microbial agents. They also provide a basis for designing and manipulating synthetic microbial communities to promote sustainable development in eco-agriculture.


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