1.Serological characteristics of individuals with hepatitis C virus/hepatitis B virus overlapping infection
Yanfei CUI ; Xia HUANG ; Chao ZHANG ; Yingjie JI ; Song QING ; Yuanjie FU ; Jing ZHANG ; Li LIU ; Yongqian CHENG
Journal of Clinical Hepatology 2026;42(1):74-79
ObjectiveTo investigate the status of overlapping hepatitis B virus (HBV) infection in patients with chronic hepatitis C virus (HCV) infection and the serological characteristics of such patients. MethodsA total of 8 637 patients with HCV infection who were hospitalized from January 1, 2010 to December 31, 2020 and had complete data of HBV serological markers were enrolled, and the composition ratio of patients with overlapping HBV serological markers was analyzed among the patients with HCV infection. The patients were divided into groups based on age and year of birth, and serological characteristics were analyzed, and the distribution of HBV-related serological characteristics were analyzed across different HCV genotypes. ResultsThe patients with HCV/HBV overlapping infection accounted for 5.85%, and the patients with previous HBV infection accounted for 48.10%; the patients with protective immunity against HBV accounted for 14.67%, while the patients with a lack of protective immunity against HBV accounted for 31.39%. The patients were divided into groups based on age: in the 0 — 17 years group, the patients with protective immunity against HBV accounted for 61.41% (304 patients); the 18 — 44 years group was mainly composed of patients with previous HBV infection (698 patients, 37.31%), the 45 — 59 years group was predominantly composed of patients with previous HBV infection (1 945 patients, 50.38%), and the ≥60 years group was also predominantly composed of patients with previous HBV infection (1 486 patients, 61.66%). The patients were divided into groups based on the year of birth: in the pre-1992 group, the patients with previous HBV infection accounted for 51.63% (4 112 patients); in the 1992 — 2005 group, the patients with protective immunity against HBV accounted for 54.72% (168 patients); in the post-2005 group, the patients with protective immunity against HBV accounted for 64.38% (235 patients). In this study, 6 301 patients underwent HCV genotype testing: the patients with genotype 1b accounted for the highest proportion of 51.71% (3 258 patients), followed by those with genotype 2a (1 769 patients, 28.07%), genotype 3b (63 patients, 1.00%), genotype 3a (10 patients, 0.16%), genotype 4 (21 patients, 0.33%), and genotype 6a (5 patients, 0.08%). ConclusionWith the implementation of hepatitis B planned vaccination program in China, there has been a significant reduction in the proportion of patients with previous HBV infection among the patients with HCV/HBV overlapping infection, but there is still a relatively high proportion of patients with a lack of protective immunity against HBV.
2.Heat-clearing and Toxin-removing Method Reduces Ischemic Stroke Injury by Protecting Endothelial-pericyte and Inhibiting Macrophage Migration
Zijin SUN ; Haojia ZHANG ; Kai WANG ; Zhaoyi WANG ; Linjing SONG ; Wenxiu XU ; Jing JI ; Changxiang LI ; Qingguo WANG ; Xueqian WANG ; Fafeng CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):56-67
ObjectiveTo investigate the regulatory effects of Huanglian Jiedutang (HLJDT) on immune cell migration, blood-brain barrier protection, and cellular functional recovery in a model of ischemic stroke. MethodsA transient middle cerebral artery occlusion (tMCAO) model was established in mice to induce ischemic stroke. Cerebral blood flow and neurological function were evaluated using laser speckle imaging and neurological deficit scoring. Histopathological damage in brain tissues was assessed by hematoxylin-eosin (HE) and Nissl staining. Mice were divided into a sham group, a model group, an HLJDT group, and a Ginkgo biloba extract (GBE) group. After one week of acclimatization, intragastric administration was initiated. The sham and model groups received normal saline, the HLJDT group received HLJDT at 1.82 g·kg-¹, and the GBE group received GBE at 0.432 g·kg-¹. Administration was continued for 5 consecutive days, and the tMCAO model was established after the final dose on day 6. Single-cell RNA sequencing was performed on brain tissues and peripheral immune cells. UMAP and odds ratio (OR) indices were used to analyze cell distribution. Differential expression analysis was conducted to evaluate the effects of HLJDT on endothelial cells, pericytes, and macrophages, combined with CellChat and decoupler to analyze cell-cell communication and transcription factor regulation. Finally, PCR and ELISA were used to validate the mRNA and protein expression of relevant genes. ResultsCompared with the sham group, the model group showed significantly increased neurological deficit scores (P<0.01) and significantly decreased cerebral blood flow (P<0.01), accompanied by cortical structural disorder, aggravated cytoplasmic vacuolization, and increased numbers of Nissl bodies. Compared with the model group, both the HLJDT and GBE groups exhibited significantly reduced neurological deficit scores (P<0.01) and markedly improved cerebral blood flow (P<0.01), along with amelioration of cortical structural disorder, alleviated cytoplasmic vacuolization, and reduced numbers of Nissl bodies. Single-cell analysis showed that HLJDT protected endothelial cells and pericytes by preventing their reduction, restored the expression of functional genes in these cells (e.g., PECAM1 and NOS3), and downregulated the expression of chemokines and adhesion-related factors (e.g., CCL2 and CXCL2). In macrophages, HLJDT reduced their recruitment to the central nervous system and downregulated the expression of chemokine receptors and inflammatory factors (e.g., IL-6, CCR2, and CXCR2). Cell-cell communication analysis further indicated that HLJDT, through the above mechanisms, alleviated damage to pericytes and endothelial cells, reduced their recruitment of macrophages, and decreased ligand-receptor interactions in chemokine signaling pathways (including CCL, CXCL, and CSF3) between pericytes/endothelial cells and macrophages, thereby preventing secondary injury. Compared with the sham group, the model group showed significantly upregulated mRNA expression levels of IL-1β, IL-6, TNF-α, CCL2, CXCL2, and CSF3 (P<0.01), while mRNA expression levels of endothelial- and pericyte function-related genes (RGS5, PECAM1, VEGFB, and NOS3) were significantly downregulated (P<0.01). In contrast, compared with the model group, the HLJDT and GBE groups exhibited significantly decreased mRNA expression levels of IL-1β, IL-6, TNF-α, CCL2, CXCL2, and CSF3 (P<0.01), and significantly increased expression of RGS5, PECAM1, VEGFB, and NOS3 (P<0.01). At the protein level, compared with the sham group, the model group showed significantly increased expression of IL-1β, IL-6, and TNF-α (P<0.01), whereas these protein levels were significantly reduced in the HLJDT and GBE groups compared with the model group (P<0.01). ConclusionHLJDT reduces neuronal damage in ischemic stroke by protecting endothelial cells and pericytes, while inhibiting their interaction with macrophages, thereby mitigating secondary injury in the central nervous system.
3.Correlation of mitochondrial genetic differentiation and spatial variables of Oncomelania hupensis robertsoni in Yunnan Province
Yuanyuan ZHANG ; Jing SONG ; Yuwan HAO ; Zaogai YANG ; Xinping SHI ; Siqi NING ; Hongqiong WANG ; Chunhong DU ; Jihua ZHOU ; Zongya ZHANG ; Kai LI ; Shizhu LI ; Yi DONG
Chinese Journal of Schistosomiasis Control 2026;38(1):54-59
Objective Objective To analyze the potential spatial factors affecting the genetic differentiation of Oncomelania hupensis robertsoni in Yunnan Province. Methods A total of 13 administrative villages were selected from schistosomiasis-endemic areas of Yunnan Province as O. hupensis snail sampling sites. At least 200 snails were collected in each site, and the spatial variable data of each site were recorded, including longitude, latitude and altitude. Thirty active and Schistosoma japonicum uninfected O. hupensis snails were selected from each sampling site by means of the crawling method and the cercarial shedding method. Genomic DNA was extracted from O. hupensis snails. Following PCR amplification, purification of PCR amplification products and sequencing, the gene sequences of O. hupensis snail samples were spliced and edited using the DNAstar software and the NCBI database to yield the complete mitochondrial sequences of O. hupensis snails at each sampling site, and the mitochondrial genetic distance matrix of O. hupensis robertsoni was calculated at each sampling site. The geographical coordinates of each sampling site were marked using the software ArcGIS 10.2, and the straight-line geographical distance between each sampling site was calculated. The altitude difference, longitude difference and latitude difference between each sampling site were calculated using the Excel software, and the correlation between the mitochondrial genetic distance matrix of O. hupensis robertsoni and each spatial variable matrix was examined by using the Mantel test at 13 sampling sites in Yunnan Province. Results Among the 13 O. hupensis snail sampling sites in Yunnan Province, the largest mitochondrial genetic distance of O. hupensis robertsoni snail populations was seen between Anding Village, Nanjian Yi Autonomous County and Caizhuang Village, Midu County (26.244 2), and the largest geographical distance was seen between Dongyuan Village, Gucheng District and Cangling Village, Chuxiong County (272.64 km). The highest altitude difference was seen between Anding Village, Nanjian Yi Autonomous County and Dongyuan Village, Gucheng District (1 086.10 m), and the largest longitude difference was found between Qiandian Village, Eryuan County and Cangling Village, Chuxiong County (1.86°), while the largest latitude difference was measured between Leqiu Village, Nanjian Yi Autonomous County and Dongyuan Village, Gucheng District (1.81°). In addition, the mitochondrial genetic distance of O. hupensis robertsoni snail populations was positively correlated with altitude at 13 snail sampling sites in Yunnan Province (r = 0.542 8, P < 0.001), and showed no significant correlations with geographical distance (r = 0.093 4, P > 0.05), longitude (r = −0.199 5, P > 0.05) or latitude (r = 0.205 7, P > 0.05). Conclusion Altitude may be a potential spatial factor affecting the genetic differentiation of O. hupensis robertsoni in Yunnan Province.
4.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.
5.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.
6.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.
7.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.
8.Surveillance of Oncomelania hupensis snails following interruption of schistosomiasis transmission in Yunnan Province
Siqi NING ; Yi DONG ; Chunhong DU ; Lifang WANG ; Yun ZHANG ; Yuhe HE ; Hua JIANG ; Jiayu SUN ; Chunqiong CHEN ; Jiaqi YAN ; Jihua ZHOU ; Zongya ZHANG ; Hongqiong WANG ; Meifen SHEN ; Jing SONG
Chinese Journal of Schistosomiasis Control 2026;38(2):200-206
Objective To investigate the distribution characteristics of Oncomelania hupensis snails in Yunnan Province fol-lowing interruption of schistosomiasis transmission, so as to provide the evidence for assessing the risk of schistosomiasis transmission and scientifically formulating the schistosomiasis surveillance program. Methods According to the requirements of the National Schistosomiasis Surveillance Scheme (2020 Edition), O. hupensis snail surveillance data were collected from 18 schistosomiasis-endemic counties (cities, districts) in Yunnan Province from 2020 to 2024, including area of snail survey, area of snail habitats, area of re-emerging snail habitats, number of frames surveyed, number of frames with O. hupensis snails, number of O. hupensis snails captured, and number of living snails, and the occurrence of frames with snails and mean density of living snails were calculated. Changes in snail status over the 5-year period from 2020 to 2024 and the differences in snail distributions specified by epidemic intensity, environmental type, and vegetation type were analyzed. Results The areas of snail survey increased from 1 727.96 hm2 in 2020 to 3 894.45 hm2 in 2024 (peak) across 18 schistosomiasis-endemic counties (cities, districts) in Yunnan Province during the period from 2020 through 2024. The areas of snail habitats increased from 70.36 hm2 in 2020 to a peak in 2023 (172.04 hm2), followed by a reduction to 132.36 hm2 in 2024, and the areas of re-emerging snail habitats increased from 42.71 hm2 in 2020 to a peak in 2022 (78.43 hm2), followed by a reduction to 40.21 hm2 in 2024. The occurrence of frames with snails and mean density of living snails increased from 1.24% (3 025/244 404) and (0.033 2 ± 0.038 7) snails/0.1 m2 in 2020 to peaks at 2.03% (6 231/307 563) and (0.066 9 ± 0.068 4) snails/0.1 m2 in 2023, followed by reductions to 1.04% (5 829/559 941) and (0.032 6 ± 0.057 7) snails/0.1 m2 in 2024, respectively. There was a significant difference in the occurrence of frames with snails over the 5-year study period (χ2 = 1 962.95, P < 0.05), and the occurrence of frames with snails reduced by 48.71% in 2024 relative to in 2023 (χ2 = 1 411.05, P < 0.005); however, there was no significant difference in the mean density of living snails over the 5 years (H = 5.310, P > 0.05). There were significant differences in the occurrence of frames with snails (χ2 = 481.27, P < 0.05) and mean density of living snails (H = 6.872, P < 0.05) in schistosomiasis-endemic areas with different epidemic intensities. The occurrence of frames with snails (χ2 = 25.32 and 38.70, both P values < 0.017) and mean density of living snails (Z = 28.55 and 49.96, both P values < 0.017) were higher in schistosomiasis transmission-interrupted and eliminated areas with snails than in schistosomiasis-eliminated areas without snails, and the occurrence of frames with snails (χ2 = 453.54, P < 0.017) and mean density of living snails (Z = −56.97, P < 0.017) were higher in schistosomiasis-eliminated areas with snails than in schistosomiasis transmission-interrupted areas with snails. O. hupensis snails were mainly distributed in paddy fields, dry farmlands and ditches; however, the occurrence of frames with snails (13.40%, 424/3 164) and mean density of living snails [(0.252 8 ± 0.158 7) snails/0.1 m2] were higher in ponds/weirs than in other types of environments (both P values < 0.05). Rice, dry farmland crops and weeds were main vegetations in which O. hupensis snails were distributed, and the occurrence of frames with snails (2.29%, 7 111/310 140) and mean density of living snails [(0.072 3 ± 0.018 9) snails/0.1 m2] were higher in weeds than in other types of environments (both P values < 0.05). Conclusions O. hupensis snails have been effectively controlled in Yunnan Province following implementation of integrated schistosomiasis control measures; however, there are still risk factors for schistosomiasis transmission, including reduced attention to schistosomiasis control and snail re-emergence. Improved control efforts and surveillance system construction and timely identification of risk factors of snail status and timely management are recommended to ensure the achievement of the target of schistosomiasis elimination as scheduled.
9.Correlation between depressive symptom and traditional Chinese medicine constitution among school aged children and adolescents
Chinese Journal of School Health 2025;46(9):1222-1225
Objective:
To explore the correlation between traditional Chinese medicine (TCM) constitution and depressive symptom among school aged children and adolescents, so as to provide evidences for informing constitution based regulation and prevention of depressive symptom.
Methods:
From June to December 2024, a total of 4 729 students aged 6-14 were recruited by cluster random sampling from 10 primary schools in Baoding (Hebei Province), Heze and Liaocheng (Shandong Province). General information, TCM constitution and depressive symptom were collected. Restricted cubic spline (RCS) models were used to analyze related factors and threshold effects of depressive symptom. Binary Logistic regression was applied to examine the association between depressive symptom and TCM constitution, with subgroup analyses conducted.
Results:
The detection rate of depressive symptom among the included children and adolescents was 25.82%. RCS analyses indicated non linear associations between depressive symptom and age (inflection point at 10 years old), bedtime (inflection point at 22:00), and wake up time (inflection point at 6:30 ) (all P non linearity <0.01). Linear associations were observed with body mass index (BMI) and sleep duration (all P non linearity > 0.05 ). After adjusting for covariates such as age, BMI and sleep status, binary Logistic regression analyses showed that Yin deficient constitution ( OR =1.26, 95% CI =1.09-1.45) and Phlegm-dampness constitution ( OR =1.42, 95% CI =1.11-1.82) were significantly associated with depressive symptom among children and adolescents (all P <0.05).
Conclusions
Depressive symptom among school aged children and adolescents is primarily associated with Yin deficiency and Phlegm dampness constitutions in TCM constitution. Active attention should be paid to susceptible TCM constitution among children and adolescents. Targeted health guidance and interventions should be implemented to improve TCM constitution health status for preventing the occurrence of depressive symptom.
10.Longitudinal cross lagged analysis of body mass index and weight stigma with depressive symptom in adolescents
DONG Ziqi, SONG Xinli, YUAN Wen, LI Jing, YANG Tian, ZHANG Xiuhong, SONG Yi, DONG Yanhui
Chinese Journal of School Health 2025;46(9):1242-1245
Objective:
To explore the bidirectional associations among body mass index Z scores (BMI Z scores) and weight stigma with depressive symptoms in adolescents, thereby providing evidence for targeted intervention strategies.
Methods:
A stratified cluster random sampling method was employed to select 18 301 adolescents aged 12-18 years from all 12 prefectures (103 counties) in the Inner Mongolia Autonomous Region, and two waves of longitudinal surveys were conducted in September 2023 (T1) and September 2024 (T2) among the adolescents. Weight stigma was assessed by using a self developed questionnaire, depressive symptom was measured with the Center for Epidemiologic Studies Depression Scale (CES-D), and BMI Z scores were calculated according to the World Health Organization standards. Pearson correlation analysis was used to examine associations among variables, and cross lagged panel models were constructed to investigate the dynamic bidirectional relationships among the three variables.
Results:
Adolescents BMI Z scores and weight stigma with depressive symptoms all exhibited autoregressive stability across the two time points (autoregressive paths, all P <0.01). Cross lagged model comparisons indicated that the bidirectional path model achieved the best fit ( χ 2=12.65, RMSEA =0.017, CFI =1.000; △ χ 2=193.39, P <0.01), supporting dynamic bidirectional associations among the three variables. After adjusting for gender, age, subjective social status and only child status, T1 BMI Z scores among adolescents positively predicted T2 weight stigma ( β =0.061), and T1 weight stigma positively predicted T2 depressive symptoms ( β =0.608); in the reverse direction, T1 depressive symptoms predicted T2 weight stigma ( β =0.003), and T1 weight stigma predicted T2 BMI Z scores ( β =0.081) (all P <0.01).
Conclusions
There is a bidirectional cross lagged relationship among adolescents BMI Z scores and weight stigma with depressive symptoms, suggesting that weight stigma may serve as a key psychological variable linking obesity and depressive symptoms. Greater attention should be paid to the potential threat of weight stigma to adolescents mental health, with intervention strategies expanded from a solely physiological focus to encompass psychosocial dimensions.


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