1.Analysis of Differential Metabolites of Pinelliae Rhizoma at Different Browning Stages Based on Widely Targeted Metabolomics
Jing TAO ; Honghong LIANG ; Ruoshi LI ; Zhouli XU ; Minzhao LI ; Aien TAO ; Guihua JIANG ; Li AI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):191-199
ObjectiveTo investigate differential metabolites associated with browning in the post-harvest processing of Pinelliae Rhizoma, providing data support for elucidating the key metabolites and metabolic pathways involved in browning, and developing safe and efficient sulfur-free processing techniques. MethodsUltra-performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry(UPLC-QTRAP-MS/MS) was used to detect the metabolites of Pinelliae Rhizoma at different browning stages(0, 8, 16 h) for widely targeted metabolomics. Subsequently, Multivariate statistical analysis of metabolites was conducted using principal component analysis(PCA), hierarchical cluster analysis(HCA), orthogonal partial least squares-discriminant analysis(OPLS-DA), and K-means cluster analysis. Differential metabolites at different browning stages were screened based on variable importance in the projection(VIP) value>1 and |log2fold change(FC)|≥1, and metabolic pathway enrichment analysis was performed using Kyoto Encyclopedia of Genes and Genomes(KEGG). ResultsA total of 1 416 metabolites were identified across the three browning stages of Pinelliae Rhizoma, predominantly comprising amino acids and their derivatives(239), lipids(219), alkaloids(156), phenolic acids(121), terpenoids(113), and flavonoids(111). A two-by-two comparison of the three browning phases, yielded 622 differential metabolites that were significantly enriched in the phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, and purine metabolic pathway. Further analysis revealed that carbohydrates such as D-mannose and turanose, phenolic acids such as 1-O-caffeoyl-6-O-glucosyl-β-D-glucose, dicaffeoylshikimic acid, and flavonoids such as epigallocatechin gallate, vitexin-7-O-rutinoside, luteolin-7-O-(6″-malonyl)glucoside-5-O-arabinoside, catechin gallate, epicatechin gallate, isovitexin-7-O-glucoside-2″-O-rhamnoside, apigenin-7-O-rutinoside-4ʹ-O-sophoroside, 3,5,3ʹ,4ʹ,5ʹ-penta-hydroxyflavan-7-gallate may act as browning substrates and play important roles in the browning process. ConclusionCarbohydrates, phenolic acids, and flavonoids may serve as key substrates in the browning process of Pinelliae Rhizoma, involving pathways such as phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, and purine metabolism, which can provide a theoretical basis for further exploration of the browning mechanism.
2.Reflections on Status Quo and Development Pathways of Traditional Chinese Medicine Technology Transfer in Context of Digital-intelligent Transformation
Jie ZHANG ; Jing XU ; Guangwei ZHENG ; Huayu ZHANG ; Chang LIU ; Xiaoxiao WEN ; Xishui PAN ; Bin WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):235-240
As a distinctive resource of Chinese civilization, traditional Chinese medicine (TCM) technology transfer faces significant opportunities under the background of digital and intelligent transformation, while also being constrained by unique challenges such as the complexity of its theoretical system, lengthy industrial chains, and multidimensional policy restrictions, resulting in a "high-value-high-threshold" paradox. At present, TCM technology transfer is deeply trapped in a "threefold reluctance" dilemma, i.e., unwillingness to transfer, inability to transfer, and lack of capacity to transfer. Specifically, the disconnection between scientific research evaluation systems and market demand leads to low conversion rates of research achievements, unclear ownership and compliance risks suppress innovation incentives, and the absence of professional services intensifies supply-demand mismatches. This article systematically analyzes the specific characteristics of TCM technology transfer and proposes a breakthrough pathway centered on full-chain digital and intelligent transformation. By integrating technologies such as intelligent sorting systems, blockchain-based traceability, and AI diagnostic models, the TCM ecosystem spanning "cultivation-production-service" can be reconstructed. In terms of standardization, promoting the progression from "experience-based data conversion" to "data standardization" and further to "intelligent standardization" is advocated to resolve quality control challenges. For example, a "three-no-one-full" certification system can strengthen quality trust. Policy coordination should focus on optimizing mechanisms for the transformation of scientific and technological achievements, while exploring intellectual property securitization and risk-sharing models to stimulate research momentum. In terms of internationalization, reliance on the Belt and Road Initiative platform to promote the export of geo-authentic medicinal material brands and standards is recommended to build a dual-driven model of "technology plus culture". Looking ahead, through the construction of national-level databases, the cultivation of interdisciplinary talent, and the mutual recognition of international standards, a new paradigm of "scientific intelligent manufacturing" can be formed, providing systematic solutions for the modernization of TCM and global health governance.
3.Cross-cultural adaptation of blood donation behavior intention scale and its reliability and validity test in college students
Yuan CHEN ; Jiao XU ; Jing XIONG ; Jing XU ; Qing ZHANG
Chinese Journal of Blood Transfusion 2026;39(1):83-89
Objective: To translate the common metrics for donation attitude, subjective norm, perceived behavioral control, and intention for the blood donation context (BD-ASPI) into Chinese, and to test its reliability and validity among college students. Methods: A research team was established. Following Beaton's cross-cultural adaptation guidelines, the BD-ASPI was translated, culturally adapted, and pre-tested to develop the Chinese version. Using convenience sampling, 620 students from four universities in Wuhan were surveyed form August to November 2024 to test the scale's reliability and validity. Results: The Chinese version of the scale consisted of 21 items across four dimensions: attitude towards blood donation, subjective norm, perceived behavioral control, and intention. The item-level content validity index ranged from 0.89 to 1.00, and the average scale-level content validity index was 0.984. Confirmatory factor analysis indicated a good fit for the second-order factor model. The Criterion validity was 0.509 (P<0.001). The overall Cronbach's α coefficient was 0.965, with the coefficients for each dimension ranging from 0.891 to 0.974. The test-retest reliability was 0.894. Conclusion: The Chinese version of the BD-ASPI demonstrates good reliability and validity, and can serve as an effective tool for assessing the behavioral intention of voluntary blood donation among college students in China.
4.Advances in the assessment of diabetic retinopathy severity in periarterial capillary-free zone by optical coherence tomography angiography
International Eye Science 2026;26(3):441-446
Diabetic retinopathy(DR), the most common microvascular complication of diabetes, has become a leading cause of visual impairment and blindness across all age groups. The early diagnosis and severity assessment of DR rely on the precise evaluation of retinal microvascular alterations. The periarterial capillary-free zone(paCFZ), a physiological avascular region surrounding retinal arteries, has recently been recognized as an important biomarker reflecting the status of retinal microcirculation. Advances in optical coherence tomography angiography(OCTA)have enabled noninvasive, high-resolution quantification of the paCFZ, offering a novel approach for the early detection and stratification of DR. This review systematically summarizes the definition and developmental mechanism of the paCFZ, as well as its morphological characteristics across different stages of DR, with a particular focus on the advantages of OCTA in visualizing and quantifying the paCFZ. We further discuss the differential manifestations of the paCFZ in nonproliferative DR and proliferative DR, and its associations with retinal ischemia and oxygenation status. In addition, the potential clinical value of paCFZ in evaluating responses to anti-vascular endothelial growth factor(VEGF)therapy and predicting disease progression is summarized. Finally, the challenges in clinical translation and future research directions are addressed, aiming to provide theoretical support and new perspectives for early screening, risk stratification, and personalized management of DR.
5.Visual analysis of hotspots and trends in global disease burden research based on CiteSpace
Jing XU ; Yuanyuan XU ; Yunshang CUI
Journal of Public Health and Preventive Medicine 2026;37(1):34-39
Objective To analyze the current status, hotspots, and trends of global disease burden research from 2015 to 2024 based on bibliometric methods, and to provide references for public health policy-making and academic research. Methods Disease burden-related literature was retrieved from the Web of Science Core Collection database, and visual analysis was conducted using CiteSpace 6.2.R4 software, including publication volume, subject distribution, national/institutional collaboration, author co-citation networks, keyword co-occurrence, clustering, and burst analysis. Results A total of 1 852 valid articles were included. The annual publication volume showed a growing trend, and entered a rapid growth phase after 2020 (with an average annual growth rate of 17.24%). The United States, China, the United Kingdom, and Germany had the highest publication volumes, with public health, internal medicine, and epidemiology being the main subject areas. The author co-citation network indicated that Younossi ZM, Lozano R, and others were core authors, while the Chinese University of Hong Kong and Aga Khan University were at the core of the institutional collaboration network. High-frequency keywords were focused on “prevalence”, “mortality”, and “risk factors”, and clustering analysis formed seven themes, including disease burden assessment methods (such as disability-adjusted life years), specific diseases (such as chronic obstructive pulmonary disease), and data resources (such as National Health and Nutrition Examination Survey). Burst analysis divided the research into three stages: 2015-2017 focused on infectious diseases (such as hepatitis C), 2018-2019 shifted to chronic non-communicable diseases (such as metabolic syndrome), and 2020-2024 focused on emerging fields such as viral-related diseases and cancer treatment. Conclusion Global disease burden research exhibits characteristics of multidisciplinary crossover. In the future, it is necessary to strengthen interdisciplinary collaboration, integrate big data and artificial intelligence technologies, focus on emerging health issues, and promote the transformation of research findings into policy.
6.Influencing Factors of Urate Crystal Deposition in Patients with Hyperuricemia and Prediction Model of TCM Syndrome Types-inflammatory Indicators
Jiaqi XU ; Bin AI ; Chao LIN ; Qiaoxuan LIN ; Changning LI ; Jing CAI ; Yan XIAO ; Jiemei GUO ; Youxin SU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):66-73
ObjectiveTo identify potential influencing factors of urate crystal deposition at ankle/foot in patients with hyperuricemia (HUA), and to analyze the predictive value of inflammatory indicators for urate crystal deposition in patients with different traditional Chinese medicine (TCM) syndromes, so as to provide potential reference for clinical risk assessment and individualized TCM intervention. MethodsA retrospective study was carried out with the enrollment of 231 HUA patients from The Third Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine between January 2021 and December 2024. The enrolled patients were further divided into a crystal deposition-positive group (143 cases) and a crystal deposition-negative group (88 cases) according to the results of dual-energy computed tomography (CT). Sociodemographic data, living habits, serum uric acid levels, and inflammatory indicators of the enrolled patients were collcted, and TCM syndrome differentiation was performed. Furthermore, univariate analysis was used to compare inter-group differences in clinical characteristics. MMultivariate Logistic regression was applied to identify the influencing factors of urate crystal deposition. In addition, the receiver operating characteristic (ROC) curves were plotted to evaluate the predictive efficacy of inflammatory indicators for crystal deposition across different TCM syndromes. ResultsThere were statistically significant inter-group differences in the proportion of males, age, body mass index, proportion of mental labor, rate of low water intake, and rate of high-sugar beverage consumption (P<0.05),whereas no significant difference in low exercise intensity was found between the two groups. Furthermore, compared with the negative group, the positive group had higher serum uric acid level, neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR), but lower systemic immune-inflammation index (SIRI) (P<0.05). Regarding the distribution of TCM syndromes, the positive group was dominated by the dampness-heat accumulation syndrome (55/143,38.46%), while the negative group was mainly characterized by the phlegm-turbidity obstruction syndrome (44/88,50.00%). Multivariate Logistic regression analysis revealed that high-sugar beverage consumption, elevated NLR, and elevated PLR were risk factors for urate crystal deposition [odd ratio (OR) = 8.002, 5.377, 1.034, respectively; 95% CI 1.572-40.732, 2.179-13.270, 1.013-1.054,all P<0.05], while SIRI was a protective factor (OR = 0.869, 95% CI 0.778-0.971, P<0.05). In the positive group, patients with the dampness-heat accumulation syndrome exhibited the highest NLR, while the lowest PLR and SIRI, showing statistically significant differences with those of other syndromes (all P<0.05). In addition, ROC curve analysis indicated that for the dampness-heat accumulation syndrome, the combined "NLR + PLR" model had an area under the curve (AUC) of 0.901 (95% CI 0.850-0.951, P<0.01), with a sensitivity of 89.1% and a specificity of 79.5%; for the blood stasis-heat obstruction syndrome, the combined "NLR + PLR" model had an AUC of 0.880 (95% CI 0.825-0.934, P<0.01), with a sensitivity of 100.0% and a specificity of 67.3%; for the liver-kidney Yin-deficiency syndrome, the single PLR model had an AUC of 0.842 (95% CI 0.731-0.952, P<0.01), with a sensitivity of 83.3% and a specificity of 84.0%. ConclusionUrate crystal deposition in HUA patients exhibits intimate associations with high-sugar beverage consumption as well as elevated NLR and PLR levels. Meanwhile, TCM syndrome differentiation has potential correlation with inflammatory characteristics. The inflammatory indicator-based prediction model constructed based on TCM syndromes exhibits good predictive value.
7.Effect of Huanglian Jiedutang on Focal Cerebral Ischemia-reperfusion Injury in Mice and Its Impact on Oligodendrocyte-related Gene Expression
Zijin SUN ; Kai WANG ; Haojia ZHANG ; Linjing SONG ; Zhaoyi WANG ; Wenxiu XU ; Jing JI ; Yonglin SHAN ; Qianqian SHI ; Xueqian WANG ; Fafeng CHENG ; Qingguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):54-63
ObjectiveTo evaluate the therapeutic effects of Huanglian Jiedutang on cerebral infarction injury in a mouse model of middle cerebral artery occlusion (MCAO) and to explore its mechanism of action on oligodendrocytes, particularly its potential in myelin repair. MethodsMultiple experimental approaches were used to evaluate cerebral ischemic injury and the effects of drug intervention. Laser speckle imaging was used to detect changes in cerebral blood flow, 2,3,5-Triphenyltetrazolium chloride (TTC) staining was used to measure infarct volume, and neurological function was scored according to the Zea-Longa criteria. Brain tissues were routinely embedded in paraffin and subjected to HE and Nissl staining to observe tissue structure and neuronal damage. Animals were divided into a sham group (n=24), model group (n=24), Huanglian Jiedutang group (n=24), and Ginkgo biloba extract (GBE) group (n=18). After 1 week of acclimatization, intragastric administration was initiated. The sham and model groups received normal saline, the Huanglian Jiedutang group was administered 1.82 g·kg-1, and the GBE group was administered 0.432 g·kg-1 after preparation as a 2.16 g·L-1 solution. All groups were treated for 5 consecutive days at a dose of 0.2 mL·(10 g)-¹·d-¹. The MCAO model was established after the final administration on day 6. Single-cell RNA sequencing was used to analyze brain tissue cellular composition and changes in oligodendrocyte subpopulations. Distinct subpopulations were identified by Uniform manifold approximation and projection (UMAP) dimensionality reduction and unsupervised clustering, and marker gene expression was analyzed. Pathway enrichment and causal inference were further performed using IPA. Finally, real-time quantitative PCR was used to verify mRNA expression changes of myelin-related genes. ResultsCompared with the sham group, the model group showed significantly increased neurological function scores (P<0.01), significantly impaired blood flow (P<0.01), significantly enlarged cerebral infarct area (P<0.01), and pathological changes including disordered cortical structural arrangement, aggravated cytoplasmic vacuolization, and increased Nissl bodies. Compared with the model group, the Huanglian Jiedutang and GBE groups showed significantly decreased neurological function scores (P<0.01), markedly restored blood flow levels (P<0.01), significantly reduced cerebral infarct area (P<0.01), and improvement in cortical structural disorder, alleviation of cytoplasmic vacuolization, and a reduction in Nissl bodies. Single-cell data showed that a myelin-associated oligodendrocyte (Mye-OL) subpopulation existed among oligodendrocytes, which was closely related to myelin generation. Compared with the sham group, the number of Mye-OL cells decreased in the model group. Compared with the model group, the number of Mye-OL cells increased in the Huanglian Jiedutang group. This subpopulation promoted the expression of myelin-related genes, including MOG, MBP, and MAG, via transcription factors such as OLIG1, OLIG2, NKX2-2, and SOX10, thereby regulating myelin generation, restoring cognition, and exerting therapeutic effects on acute cerebral infarction. Compared with the sham group, the mRNA expression levels of OLIG1, OLIG2, NKX2-2, and SOX10 were significantly downregulated in the model group (P<0.01), and the mRNA expression levels of myelin-related genes, including MOG, MBP, and MAG, were also significantly downregulated (P<0.01). In contrast, compared with the model group, the Huanglian Jiedutang and GBE groups showed significantly upregulated mRNA expression levels of OLIG1, OLIG2, NKX2-2, and SOX10 (P<0.01), and significantly upregulated mRNA expression levels of myelin-related genes, including MOG, MBP, and MAG (P<0.01). ConclusionHuanglian Jiedutang exerts therapeutic effects on acute cerebral infarction by regulating the OLIG1/2-NKX2-2-SOX10 signaling pathway to promote myelin generation by Mye-OL cells.
8.Regulatory Role of Huanglian Jiedutang in Microglial Metabolic Reprogramming to Suppress Neuroinflammatory Damage Based on Single-cell Transcriptomics
Zijin SUN ; Haojia ZHANG ; Kai WANG ; Linjing SONG ; Chuanzun WANG ; Wen WANG ; Jing JI ; Zhaoyi WANG ; Wenxiu XU ; Qingguo WANG ; Xueqian WANG ; Fafeng CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):64-73
ObjectiveTo investigate the characteristics of metabolic reprogramming during cerebral ischemia-reperfusion injury using single-cell transcriptome sequencing, analyze the heterogeneity of microglial populations, and evaluate the interventional effects of Huanglian Jiedutang on metabolic abnormalities and neuroinflammation. MethodsA transient middle cerebral artery occlusion (tMCAO) model was used to establish ischemic stroke in mice. Local cerebral blood flow changes were monitored by laser speckle imaging. Neurological impairment was evaluated using the Zea-Longa score, and histopathological damage in brain tissue was observed by HE and Nissl staining. Animals were divided into a sham group, model group, Huanglian Jiedutang group, and Ginkgo biloba extract (GBE) group. After 1 week of acclimatization, intragastric administration was initiated. The sham and model groups received normal saline, the Huanglian Jiedutang group was administered 1.82 g·kg-1, and the GBE group was administered 0.432 g·kg-1 after preparation as a 2.16 mg/mL solution. All groups were treated for 5 consecutive days (0.2 mL/10 g/day), and the tMCAO model was established on day 6 after the final administration. At the molecular level, single-cell RNA sequencing was performed on ischemic hemisphere tissue. Non-negative matrix factorization (NMF) was used to cluster microglial subpopulations, combined with differential expression analysis, metabolic reprogramming assessment, and inflammatory factor correlation analysis to elucidate their functional characteristics in ischemia-reperfusion injury. Transcription factor enrichment analysis was further conducted to identify key regulatory nodes. Finally, PCR was used to detect mRNA expression changes of relevant genes to validate the single-cell sequencing results. ResultsCompared with the sham group, the model group showed increased neurological function scores (P<0.01), decreased blood flow levels (P<0.01), disordered cortical structure, increased cytoplasmic vacuolization, and increased Nissl bodies. Compared with the model group, the Huanglian Jiedutang and GBE groups showed decreased neurological function scores (P<0.01), increased blood flow levels (P<0.01), alleviated cortical structural disorder, reduced cytoplasmic vacuolization, and decreased Nissl bodies. Single-cell analysis showed that microglia could be divided into five subpopulations. Among them, clusters 3 and 5 exhibited significant pro-inflammatory phenotypes, with marked activation of hypoxia and NF-κB signaling pathways, and were identified as pro-inflammatory subpopulations. Clusters 1 and 2 were enriched in Wnt/β-catenin and transforming growth factor(TGF)-β signaling pathways and exhibited prominent anti-inflammatory and reparative characteristics. Meanwhile, glycolysis-related genes, such as HK2, PFKP, and LDHA, were significantly upregulated in the pro-inflammatory subpopulations. Correlation analysis showed that the expression levels of inflammatory molecules were positively correlated with glycolysis-related gene expression levels, whereas the expression levels of reparative and anti-inflammatory molecules were negatively correlated with glycolysis-related gene expression levels, indicating that microglia rely on the glycolytic pathway for energy acquisition under ischemic conditions. Further single-cell transcriptome analysis revealed that Huanglian Jiedutang effectively downregulated key genes driving metabolic reprogramming (such as HK2, PFKP, and LDHA), significantly reduced the proportion of microglial subpopulations accompanied by glycolytic reprogramming, and inhibited their transformation toward a damage phenotype, thereby reducing inflammatory injury. Meanwhile, compared with the sham group, the mRNA expression levels of interleukin (IL)-1β, IL-6, tumor necrosis factor(TNF)-α, CCL2, CXCL2, and CSF3 were significantly upregulated (P<0.01) in the model group, whereas the mRNA expression levels of endothelial- and pericyte-related functional genes, including RGS5, PECAM1, VEGFB, and NOS3, were significantly downregulated (P<0.01). In contrast, compared with the model group, the Huanglian Jiedutang and GBE groups showed significantly decreased mRNA expression levels of IL-1β, IL-6, TNF-α, CCL2, CXCL2, and CSF3 (P<0.01), and significantly increased mRNA expression levels of endothelial- and pericyte-related functional genes, including RGS5, PECAM1, VEGFB, and NOS3 (P<0.01). ConclusionHuanglian Jiedutang exerts neuroprotective effects by regulating the metabolic reprogramming state of microglia and modulating their inflammatory levels, thereby inhibiting neuroinflammatory injury.
9.Volatile Component Differences in Xihuangwan Prepared with Natural and Artificial Musk Based on Non-targeted and Targeted Metabolomics
Jing WANG ; Fangzhu XU ; Li MENG ; Qizhen ZHU ; Huanjun ZHAO ; Caina YU ; Xuelian CHEN ; Hui GAO ; Zimin YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):194-201
ObjectiveHeadspace solid-phase microextraction-gas chromatography-mass spectrometry(HS-SPME-GC-MS) and GC-triple quadrupole MS(GC-QqQ-MS) in combination with non-targeted and targeted metabolomics were employed to systematically analyze the chemical composition differences of Xihuangwan prepared with natural musk and artificial musk, and establish an identification system for them. MethodsThe volatile components of 9 batches of Xihuangwan samples from 8 manufacturers were analyzed by HS-SPME-GC-MS non-targeted metabolomics, and identified by comparing their MS data with the National Institute of Standards and Technology(NIST) spectral library. Orthogonal partial least squares-discriminant analysis(OPLS-DA) was used to identify differential volatile components of Xihuangwan prepared with natural musk and artificial musk. Additionally, GC-QqQ-MS targeted metabolomics was applied to quantify the levels of α-pinene, β-elemene, muscone, dehydroepiandrosterone, bornyl acetate, and octyl acetate in 27 batches of samples from 9 manufacturers. Cluster analysis, principal component analysis(PCA), and partial least squares-discriminant analysis(PLS-DA) were conducted to further explore the differences in volatile components between Xihuangwan samples prepared with natural musk and artificial musk. ResultsNon-targeted metabolomics identified 291 volatile compounds in Xihuangwan, including alkanes, esters, alkanes, alcohols, ketones, naphthalenes and others. OPLS-DA analysis revealed distinct separation between Xihuangwan samples containing artificial musk(A1, C1, D1, E1, F1, G1, I1) and those containing natural musk(H1, H3). A total of 30 differential metabolites were identified. The relative contents of these 30 differential metabolites were visualized using a radar chart, revealing significant differences in the levels of octanol, borneol acetate and muscone. Cluster analysis and PCA results from targeted metabolomics indicated that Xihuangwan could be classified into two distinct groups:one composed of natural musk(H1, H3) and the other of artificial musk, sample H2. PLS-DA identified muscone, octyl acetate, and dehydroepiandrosterone as key differential volatile components. Although no significant difference was observed in the content of octyl acetate between the two groups, statistically significant differences were found for muscone and dehydroepiandrosterone(P<0.05). ConclusionMuscone and dehydroepiandrosterone can be used for the differentiation of Xihuangwan samples containing natural musk from those containing artificial musk. This study systematically and comprehensively analyzed the differences in the types and contents of major volatile components in Xihuangwan prepared with natural musk and artificial musk, providing a scientific basis for quality evaluation and control of Xihuangwan.
10.Effect of Huanglian Jiedutang on Focal Cerebral Ischemia-reperfusion Injury in Mice and Its Impact on Oligodendrocyte-related Gene Expression
Zijin SUN ; Kai WANG ; Haojia ZHANG ; Linjing SONG ; Zhaoyi WANG ; Wenxiu XU ; Jing JI ; Yonglin SHAN ; Qianqian SHI ; Xueqian WANG ; Fafeng CHENG ; Qingguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):54-63
ObjectiveTo evaluate the therapeutic effects of Huanglian Jiedutang on cerebral infarction injury in a mouse model of middle cerebral artery occlusion (MCAO) and to explore its mechanism of action on oligodendrocytes, particularly its potential in myelin repair. MethodsMultiple experimental approaches were used to evaluate cerebral ischemic injury and the effects of drug intervention. Laser speckle imaging was used to detect changes in cerebral blood flow, 2,3,5-Triphenyltetrazolium chloride (TTC) staining was used to measure infarct volume, and neurological function was scored according to the Zea-Longa criteria. Brain tissues were routinely embedded in paraffin and subjected to HE and Nissl staining to observe tissue structure and neuronal damage. Animals were divided into a sham group (n=24), model group (n=24), Huanglian Jiedutang group (n=24), and Ginkgo biloba extract (GBE) group (n=18). After 1 week of acclimatization, intragastric administration was initiated. The sham and model groups received normal saline, the Huanglian Jiedutang group was administered 1.82 g·kg-1, and the GBE group was administered 0.432 g·kg-1 after preparation as a 2.16 g·L-1 solution. All groups were treated for 5 consecutive days at a dose of 0.2 mL·(10 g)-¹·d-¹. The MCAO model was established after the final administration on day 6. Single-cell RNA sequencing was used to analyze brain tissue cellular composition and changes in oligodendrocyte subpopulations. Distinct subpopulations were identified by Uniform manifold approximation and projection (UMAP) dimensionality reduction and unsupervised clustering, and marker gene expression was analyzed. Pathway enrichment and causal inference were further performed using IPA. Finally, real-time quantitative PCR was used to verify mRNA expression changes of myelin-related genes. ResultsCompared with the sham group, the model group showed significantly increased neurological function scores (P<0.01), significantly impaired blood flow (P<0.01), significantly enlarged cerebral infarct area (P<0.01), and pathological changes including disordered cortical structural arrangement, aggravated cytoplasmic vacuolization, and increased Nissl bodies. Compared with the model group, the Huanglian Jiedutang and GBE groups showed significantly decreased neurological function scores (P<0.01), markedly restored blood flow levels (P<0.01), significantly reduced cerebral infarct area (P<0.01), and improvement in cortical structural disorder, alleviation of cytoplasmic vacuolization, and a reduction in Nissl bodies. Single-cell data showed that a myelin-associated oligodendrocyte (Mye-OL) subpopulation existed among oligodendrocytes, which was closely related to myelin generation. Compared with the sham group, the number of Mye-OL cells decreased in the model group. Compared with the model group, the number of Mye-OL cells increased in the Huanglian Jiedutang group. This subpopulation promoted the expression of myelin-related genes, including MOG, MBP, and MAG, via transcription factors such as OLIG1, OLIG2, NKX2-2, and SOX10, thereby regulating myelin generation, restoring cognition, and exerting therapeutic effects on acute cerebral infarction. Compared with the sham group, the mRNA expression levels of OLIG1, OLIG2, NKX2-2, and SOX10 were significantly downregulated in the model group (P<0.01), and the mRNA expression levels of myelin-related genes, including MOG, MBP, and MAG, were also significantly downregulated (P<0.01). In contrast, compared with the model group, the Huanglian Jiedutang and GBE groups showed significantly upregulated mRNA expression levels of OLIG1, OLIG2, NKX2-2, and SOX10 (P<0.01), and significantly upregulated mRNA expression levels of myelin-related genes, including MOG, MBP, and MAG (P<0.01). ConclusionHuanglian Jiedutang exerts therapeutic effects on acute cerebral infarction by regulating the OLIG1/2-NKX2-2-SOX10 signaling pathway to promote myelin generation by Mye-OL cells.


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