1.Clinical Observation on Prevention of Recurrence of Common Bile Duct Stones After ERCP with Yuyin Lidan Granules
Xiao WANG ; Yong FANG ; Cong HE ; Jiali ZHANG ; Meng YU ; Jing KONG ; Yi JIANG ; Chuanqi CHENG ; Xiaosu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):159-166
ObjectiveTo observe the clinical efficacy and safety of Yuyin Lidan granules (YYLD) in preventing the recurrence of common bile duct stones (CBDS) in patients with liver and gallbladder dampness-heat syndrome following endoscopic retrograde cholangiopancreatography (ERCP). MethodsThis randomized, parallel, controlled trial enrolled postoperative CBDS-ERCP patients who met the inclusion and exclusion criteria. Sixty-four patients were randomly assigned to an observation group or a control group, with 32 cases in each. Both groups received conventional Western medical treatment after ERCP, while the observation group additionally received YYLD for 8 weeks. The follow-up period lasted for 1 year. The efficacy indicators included bile bilirubin levels, traditional Chinese medicine (TCM) syndrome scores, clinical efficacy rate, pancreatitis and inflammation markers, postoperative liver function, and CBDS recurrence rate at 1-year follow-up, which were used to jointly evaluate the clinical efficacy and safety of both groups. ResultsA total of 56 patients completed the study and were included in the final analysis, i.e., 29 in the observation group and 27 in the control group. Baseline characteristics were comparable between the two groups. Compared with pre-treatment and with the control group after treatment, the bile bilirubin level in the observation group significantly decreased (P<0.05). After treatment, the clinical cure and marked improvement rates were higher in the observation group than in the control group, showing a statistically significant difference in overall clinical efficacy (P<0.05). Compared with pre-treatment, the primary and secondary symptoms in the observation group, as well as the primary symptom and the secondary symptom of nausea and vomiting in the control group (weeks 4 and 8), were significantly reduced (P<0.05). Compared with the control group after treatment, the observation group showed significant reductions in the primary symptom of loose stools/constipation (day 5 and week 4) and in three secondary symptoms, i.e., bitter taste and sticky dry mouth, abdominal distension and poor appetite (throughout the treatment period), and general heaviness and fatigue (day 5 and week 4), with statistical differences (P<0.05). Compared with pre-treatment, both groups showed decreased lipase and urinary amylase levels (P<0.05). However, no significant between-group differences were observed in pancreatitis or inflammation-related indices after treatment. Compared with pre-treatment, all liver function indicators in the observation group and alanine aminotransferase ( ALT ), γ-glutamyl transferase ( γ-GT ), alkaline phosphatase (ALP), and conjugated bilirubin in the control group significantly decreased at weeks 4 and 8 (P<0.05). Compared with the control group after treatment, only serum total bilirubin and unconjugated bilirubin were significantly reduced in the observation group during the treatment period (P<0.05). ConclusionYYLD combined with conventional Western medical treatment can effectively regulate bilirubin metabolism (in bile and serum), improve TCM clinical symptoms, and prevent CBDS recurrence after ERCP in patients with liver and gallbladder dampness-heat syndrome. This regimen is safe and effective and is worthy of further clinical research and promotion.
2.Herbal Textual Research on Cynanchi Atrati Radix et Rhizoma in Famous Classical Formulas
Xiaoqi JING ; Minna GUO ; Haihua WANG ; Juan LI ; Fusheng ZHANG ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):208-216
This article systematically reviews and verifies the name, origin, production area, quality evaluation, harvesting, processing and other aspects of Cynanchi Atrati Radix et Rhizoma(CARR) by consulting relevant ancient and modern literature, in order to provide a basis for the development and utilization of famous classical formulas containing this herb. Through textual research, Baiwei has been the official name for CARR, though it also bears alternative names such as Chuncao, Popo Zhenxianbao, Longdan Baiwei. The mainstream base is the roots and rhizomes of Cynanchum atratum. Historical records indicate primary producing areas include Shandong, Anhui, Jiangsu, Shaanxi and Shanxi. Since the late Ming dynasty, varieties from Juxian, Yishui and Rizhao in Shandong have been highly regarded as authentic, commonly known as eastern Baiwei. Since modern times, its quality has been summarized as fine, slender, and straight fibrous roots, pale yellow exterior, whiter interior, and dryness with easy breakability are considered superior. The harvesting time before the Song dynasty was on the third day of the third lunar month, but after the Song dynasty, harvesting was possible in both spring and autumn. The initial processing methods of CARR in ancient times included drying in the shade, removing Lu(the little rhizomes which are on tap of roots), and removing mustaches, modern methods involve washing and sun-drying. During the Northern and Southern dynasties, processing methods included steaming. In the Song dynasty, drying and light stir-frying were predominant, while wine washing emerged in the Ming dynasty. Modern practices primarily involve using raw, stir-frying or honey processing. Regarding the medicinal properties of CARR, both ancient and modern texts agree it has a bitter and salty taste and is non-toxic. Records prior to the Qing dynasty predominantly describe its nature as extremely cold, while mainstream herbal texts after the Qing dynasty generally characterize it as cold. Before the Ming dynasty, there were no records of its meridian tropism. It was not until the Qing dynasty that it was recorded in the lung meridian. Modern records mainly refer to the stomach, liver, and kidney meridians. Throughout history, its main functions have been to clear heat, diuresis, nourish Yin, and replenish essence, primarily treating Yin deficiency and fever syndrome. Based on the research results, it is suggested that when developing famous classical formulas containing CARR, the dried roots and rhizomes of C. atratum can be selected as its medicinal source. If there are no specific processing requirements, raw products can be selected as medicine. If the processing requirements are specified, corresponding processed products can be selected as medicine according to the original formula requirements.
3.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.
4.Candida albicans promotes malignant progression of oral leukoplakia: a preliminary study based on clinical cohort and animal models
CHENG Fangbo ; ZHANG Shiyu ; WANG Ying ; LI Jing
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(3):237-245
Objective:
To explore the correlation between Candida albicans and the development of oral leukoplakia (OLK), and to provide a basis for improving the pathogenic mechanism of the malignant transformation of OLK.
Methods:
Oral microbiome data were obtained from public databases (NCBI BioProject, PRJNA788378; GEO, GSE227919), and bioinformatic methods were employed to evaluate the correlation between Candida albicans infection and OLK. Approval was obtained from the institutional Medical Ethics Committee. A tissue microarray was constructed using samples collected from an OLK clinical cohort. Hematoxylin and eosin (H&E) staining and periodic acid-Schiff (PAS) staining were performed to analyze the relationship between the Candida albicans detection rate and clinicopathological features. Approval was obtained from the institutional Animal Ethics Committee. A mouse model was established by combining 4-nitroquinoline-1-oxide (4NQO) in drinking water with oral inoculation of Candida albicans (4NQO + Candida albicans group), while mice treated with 4NQO in drinking water and PBS served as the control group (4NQO + PBS group). The degree of epithelial dysplasia was compared between the two groups to assess the impact of Candida albicans infection on lesion progression (defined in this study as the progression from mild/moderate epithelial dysplasia to severe dysplasia/carcinoma in situ or invasive squamous cell carcinoma).
Results:
Bioinformatic analysis revealed that the detection rate of Candida albicans in OPMDs and OLK tissues was significantly higher than that in the healthy control group. Staining results of clinical samples demonstrated that Candida albicans colonized OLK lesions; compared with Candida albicans-negative patients, positive patients exhibited a state of high-grade progression. Animal experiments indicated that, compared with the 4NQO + PBS group, the degree of oral epithelial dysplasia in the 4NQO + Candida albicans group was significantly exacerbated, and the malignant transformation rate was higher, suggesting that Candida albicans promotes the high-grade progression of OLK.
Conclusion
Candida albicans exhibits a increasing trend during the malignant progression of the OLK. It aggravates the degree of epithelial dysplasia in OLK and promotes its transformation into high-grade lesions, suggesting that Candida albicans plays a crucial promoting role in the high-grade progression of OLK.
5.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.
6.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.
7.Herbal Textual Research on Piperis Longi Fructus in Famous Classical Formulas
Haihua WANG ; Xiaoqi JING ; Juan LI ; Dabang REN ; Fusheng ZHANG ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):210-219
This article systematically analyzes the historical evolution of the name, origin, medicinal parts, producing area, harvesting and processing, nature, flavor and efficacy of Piperis Longi Fructus by referring to the materia medica, medical books, and prescription books of past dynasties, combined with the relevant modern literature, in order to provide a basis for the development and utilization of famous classical formulas containing this herb. According to the herbal textual research, the name of Piper longum first appeared in Nanfang Caomuzhuang, and it also has other aliases such as Biboli, Halou, and Hujiaohua. Historically, the origin of Piperis Longi Fructus has been P. longum of the Piperaceae family. In ancient times, both the fruit and root were used as medicine, and since the Republic of China, the fruit has been mainly used as medicine. The medicinal part is the dried, nearly ripe or ripe fruit spikes. Piperis Longi Fructus is native to India and has been introduced into China since the Tang dynasty. In the Ming dynasty, Bencao Pinhui Jingyao clearly stated that the genuine producing area was "Duanzhou", present-day Zhaoqing in Guangdong province. Nowadays, it is planted in Guangdong, Guangxi, Hainan, Yunnan and other regions. Historically and currently, harvesting occurs in autumn. The ancient processing method uniformly involved removing the stems, soaking in the sourest vinegar overnight, baking, and scraping off the peels and grains with a knife until clean. In modern times, impurities are removed, and it is dried in the sun and crushed when used. The properties, functions and applications of P. longum are basically the same in ancient and modern times. It tastes pungent, is warm in nature, and non-toxic. It has the effects of warming the middle-jiao to dispel cold, lowering Qi and relieving pain, and is used for cold pain in the epigastrium and abdomen, vomiting, diarrhea, chest pain, headache, and toothache. Based on the research results, it is recommended that when developing famous classical formulas containing Piperis Longi Fructus, the dried nearly ripe or ripe fruit spikes of P. longum should be used. If there are no clear processing requirements, it is recommended to use the raw products for medicinal use, and the specific processing methods can refer to the relevant requirements under Piperis Longi Fructus in the 2025 edition of the Pharmacopoeia of the People's Republic of China. If processing requirements such as soaking in vinegar and peeling are clearly specified, it is recommended to follow the ancient methods.
8.Research progress on the comorbidity mechanism of dry eye and depression
Yunfan ZHANG ; Kang WANG ; Jing LI
International Eye Science 2026;26(4):629-635
Dry eye disease(DED)and depression(DEP), though anatomically and clinically distinct, show significant epidemiological, pathophysiological, and prognostic interplay. Their co-occurrence has risen sharply in recent years, yet the mechanisms driving this comorbidity remain under-investigated. This review systematically synthesizes current evidence, highlighting that pro-inflammatory cytokines, neuro-regulatory imbalance, mitochondrial dysfunction, and gut-microbiota dysbiosis constitute a shared molecular network, while sleep deprivation and antidepressant use further amplify the vicious cycle. By identifying limitations in existing studies, this review proposes future research directions to offer new theoretical and clinical insights for managing DED-DEP comorbidity.
9.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.
10.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.


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