1.Overcoming organ shortage: key clinical application technology breakthroughs and future prospects in xenogeneic organ transplantation
Junze CHEN ; Cheng ZHANG ; Yongyuan JIAN ; Kun DONG ; Shuaijun MA ; Chunqiang DONG
Organ Transplantation 2026;17(3):393-404
The technology of xenogeneic organ transplantation, as one of the core strategies to address the current contradiction between the supply and demand of transplant organs, has achieved significant breakthroughs from basic research to clinical application driven by factors such as the innovation of gene modification technology, the injection of research capital and the expansion of clinical trials, especially with the first actual clinical application of a pig heart-to-human transplant. China is also at the forefront of this field. This article intends to summarize the international research trends of xenogeneic organ transplantation (including financial support, the evolution of research stages and global clinical trial cases), and analyze the evolution and optimization of xenogeneic transplantation immunosuppression schemes, as well as the breakthroughs and unresolved scientific issues in current key clinical application technologies. The aim is to comprehensively present the progress of this field from basic research to clinical transformation, and provide references for promoting the rapid development of China's xenogeneic transplantation technology and subsequent clinical transformation and research directions.
2.Heartbeat-evoked responses to cue-induced craving in heroin use disorder individuals
Dingming CHANG ; Yongxin CHENG ; Juan WANG ; Ruowan LI ; Fang DONG ; Kai YUAN ; Dahua YU
Chinese Journal of Clinical Medicine 2026;33(2):230-239
Objective To explore the differences in heartbeat-evoked response (HER) under drug-related cues and neutral cues in individuals with heroin use disorder (HUD), and analyze the correlation between HER potentials and immediate cue-induced craving scores. Methods Fifty HUD participants were recruited from the Chang’an Compulsory Isolation Drug Rehabilitation Center in Shaanxi Province from June to September 2024. Simultaneous acquisition of 64-channel electroencephalography (EEG) and electrocardiogram signals was performed. Twenty alternating segments of drug-related and neutral cue videos were presented, and participants rated their subjective craving after each segment using visual analogue scale (VAS) scores. Scalp EEG data were source analyzed to obtain cortical EEG signals and corresponding HER. Short-time Fourier transform was used to calculate the power spectral density (PSD) of EEG within a time window from 100 ms before the R-peak to 500 ms after it, using the R-peak as the time zero point. Cluster-based permutation testing was used to analyze PSD differences between drug-related and neutral cues in the HUD individuals. Pearson correlation analysis was performed to evaluate the correlation between HER potentials and VAS scores. Results In the 350–420 ms time window, HER potentials in the left posterior parietal, temporal, and posterior cingulate cortices were significantly lower under drug-related cues compared to neutral cues (P<0.01); in the 140–210 ms time window, HER potentials in the right prefrontal cortex were significantly higher under drug-related cues compared to neutral cues (P<0.01). Correlation analysis showed that HER potentials in the left temporal and left posterior cingulate cortices were significantly negatively correlated with VAS scores (P<0.05). Drug-related cues enhanced PSD of γ power (30–100 Hz) in salience network (fronto-insular), parietal and occipital regions (P<0.05). PSD integrations of low-γ power (40–60 Hz) in parietal region (350–400 ms) and high-γ power (70–100 Hz) in left salience network (fronto-parietal) and occipital regions (300–350 ms) were positively correlated with VAS scores (P<0.05). Conclusions Drug-related cues may modulate cortical activity related to heartbeat perception in HUD individuals, and such dynamic changes in both time and frequency domains are stably associated with subjective craving.
3.Empirical study of input, output, outcome and impact of community-based rehabilitation stations
Xiayao CHEN ; Ying DONG ; Xue DONG ; Zhongxiang MI ; Jun CHENG ; Aimin ZHANG ; Didi LU ; Jun WANG ; Jude LIU ; Qianmo AN ; Hui GUO ; Xiaochen LIU ; Zefeng YU
Chinese Journal of Rehabilitation Theory and Practice 2026;32(1):83-89
ObjectiveTo investigate the present situation of input, output, outcome and impact of all registered community-based rehabilitation stations in Inner Mongolia in China, and analyze how the input predict the output, outcome and impact. MethodsFrom March 1st to April 30th, 2025, a questionnaire survey was conducted on all registered community-based rehabilitation stations in Inner Mongolia, covering four dimensions: input, output, outcome and impact. A total of 1 365 questionnaires were distributed. The input included four items: laws and policies, human resources, equipment and facilities, and rehabilitation information management. The output included two items: technical paths and benefits/effectiveness. The outcome included three items: coverage rates, rehabilitation interventions and functional results. The impact included two items: health and sustainability. Each item contained several questions, all of which were described in a positive way. Each question was scored from one to five. A lower score indicated that the situation of the community-based rehabilitation station was more in line with the content described in the question. Regression analysis was performed using the total score of each item of input dimension as independent variables, and the total scores of the output, outcome and impact dimensions as dependent variables. ResultsA total of 1 262 valid questionnaires were collected. The mean values of input, output, outcome and impact of community-based rehabilitation stations were 1.827 to 1.904, with coefficient of variation of 45.892% to 49.239%. The regression analysis showed that, rehabilitation information management, human resources, and laws and policies significantly predicted the output dimension (R² = 0.910, P < 0.001). Meanwhile, all four items in the input dimension predicted both the outcome (R² = 0.850, P < 0.001) and impact dimensions (R² = 0.833, P < 0.001). ConclusionInput, output, outcome and impact of the community-based rehabilitation stations in Inner Mongolia were generally in line with the content of the questions, although some imbalances were observed. Additionally, the input of community-based rehabilitation stations could significantly predict their output, outcome and impact.
4.Clinical characteristics and prognosis of immunotherapy for recurrent/metastatic nasopharyngeal carcinoma: a single-center retrospective analysis
WANG Haoqiang ; LIU Baiyang ; YANG Ning ; LIU Peng ; CHENG Donghai ; PENG Lijun ; WANG Xianci ; HUANG Xueqin ; DONG Enlai ; JIANG Yiming ; ZHOU Juan ; XIE Bo
Chinese Journal of Cancer Biotherapy 2026;33(1):84-90
[摘 要] 目的:探讨复发/转移性鼻咽癌(NPC)接受含PD-1单抗免疫治疗的临床特征和预后影响因素。方法:回顾性分析2019年3月至2024年7月期间南部战区总医院确诊的95例NPC患者的临床资料和外周血生化及免疫学指标。预后分析采用Kaplan-Meier曲线,组间比较使用Log-rank检验,采用Cox比例风险模型进行单因素和多因素分析。结果:95例患者中男性81例,女性14例,中位年龄49.72岁(16~74岁),Ⅳ期91例(95.79%),所有患者均采用免疫治疗,联合或不联合化疗方案治疗,中位无进展生存期(mPFS)为10.5个月,客观缓解率(ORR)70.53%,疾病控制率(DCR)89.47%,接受含铂治疗方案患者PFS相对更长,且差异有统计学意义。紫杉醇 + 顺铂 + 氟尿嘧啶(TPF)对比吉西他滨 + 顺铂(GP)和紫杉醇 + 顺铂(TP)显示出更长的PFS,但差异无统计学意义。不同PD-1单抗治疗组间的PFS未显示出有统计学意义的差异。单因素及多因素Cox回归分析结果显示,肿瘤复发状态、初始血浆EBV感染状态、治疗周期数、基线外周血SII是复发/转移性NPC患者接受PD-1抑制剂治疗疗效预测的独立相关因素(均P < 0.05),并且非复发患者、初始血浆EBV DNA阳性、接受 ≥ 4治疗周期、基线外周血SII < 772.81的患者接受PD-1抑制剂治疗预后相对更好。结论:在接受PD-1抑制剂治疗的复发/转移性NPC患者中,非复发患者、初始血浆EBV DNA阳性、≥ 4治疗周期且外周血SII < 772.81者PFS相对更长,可早期识别免疫治疗效果不佳患者并精准干预。
5.Tracing Development of LIU Wansu's Theory of ''Fire-heat Inducing Stroke''
Xin LAN ; Changxiang LI ; Haojia ZHANG ; Jialin CHENG ; Zijin SUN ; Liyang DONG ; Zilin REN ; Xueqian WANG ; Fafeng CHENG ; Qingguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):32-41
LIU Wansu, as the foremost of the four great masters of the Jin-Yuan period, established the "theory of fire-heat'' and extended the fire-heat pathogenesis framework to the field of stroke, thereby forming the theory of ''fire-heat inducing stroke''. This achieved a paradigmatic shift in stroke etiology from ''exogenous wind inducing stroke'' to ''fire-heat inducing stroke''. This paper systematically reviews the developmental trajectory of LIU Wansu's ''fire-heat inducing stroke'' theory and explores the social background, academic origins, and core connotations of its theoretical construction. The study found that, based on the ''Nineteen Pathomechanisms'' in the Huangdi's Internal Classic (Huang Di Nei Jing) and combined with clinical practice, LIU Wansu proposed that fire-heat is the fundamental cause of stroke, and that the Six Climatic Factors and the Five Zhi-Emotions can all transform into fire. He further constructed a stratified syndrome differentiation and therapeutic system centered on clearing heat and purging fire, emphasizing differentiated treatment of exterior and interior syndromes, Six Meridians syndrome differentiation, and seasonally adjusted medication. This theory not only resolved the diagnostic and therapeutic dilemmas of febrile epidemic diseases during the Jin-Yuan period, but also exerted a profound influence on later physicians such as ZHANG Zihe and ZHU Danxi, thereby promoting the pluralistic development of stroke theory in traditional Chinese medicine (TCM). Modern pharmacological research provides solid scientific evidence, confirming that the ''fire-heat'' pathological state is highly associated with key mechanisms such as excessive inflammatory responses, oxidative stress, and excitatory amino acid toxicity following cerebral ischemia. Heat-clearing and fire-purging prescriptions and agents, such as Huanglian Jiedu Tang and baicalin, can exert multi-target neuroprotective effects by regulating inflammatory signaling, enhancing antioxidant enzyme activity, and balancing neurotransmitters. This not only verifies the scientific basis of the ''fire-heat inducing stroke'' theory from a modern biological perspective but also provides conclusive evidence for the clinical application of heat-clearing and fire-purging therapy. LIU Wansu's ''fire-heat inducing stroke'' theory represents a major milestone in the historical understanding of stroke pathogenesis, and its academically transitional insights continue to hold core guiding value for the pattern identification and treatment of ischemic stroke today.
6.Effects of Huanglian Jiedutang on Neutrophil Infiltration in Brain of MCAO Mice via Regulation of Chemokine Expression in Exosomes
Haojia ZHANG ; Kai WANG ; Zijin SUN ; Chunyu WANG ; Wei SHAO ; Kunjing LIU ; Liyang DONG ; Dan CHEN ; Wenxiu XU ; Chuanzun WANG ; Wen WANG ; Changxiang LI ; Xueqian WANG ; Fafeng CHENG ; Qingguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):42-53
ObjectiveTo investigate whether Huanglian Jiedutang can inhibit neutrophil infiltration in the brains of middle cerebral artery occlusion (MCAO) mice by regulating the expression of neutrophil-related chemokines in exosomes, thereby achieving therapeutic effects. MethodsA total of 130 male specific pathogen-free (SPF) C57BL/6J mice were randomly divided into four groups: Sham-operated group, MCAO model group, Huanglian Jiedutang group (6 g·kg-1), and Ginaton group (21.6 mg·kg-1), with 10 mice in the Ginaton group and 40 mice in each of the remaining three groups. Mice in the Huanglian Jiedutang group and the Ginaton group were administered the corresponding drugs by oral gavage once daily at a volume of 0.15 mL·(10 g)-1 for 7 consecutive days, while the sham-operated and model groups received an equal volume of saline via the same route. After 7 days, MCAO surgery was performed. The distal and proximal ends of the right common carotid artery (CCA) were ligated, a small incision was made between the two ligatures, and a silicone rubber-coated monofilament with a rounded tip was inserted into the lumen to occlude the CCA. The filament was left in place for 1 h to establish a focal cerebral ischemia model. At 24 h after modeling, mice were evaluated. Neurological function was assessed using the Longa score. Cerebral infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Cerebral blood flow was observed by laser speckle imaging. Hematoxylin and eosin (HE) staining and Nissl staining were used to observe pathological changes in brain tissues. Exosomes were isolated from mouse plasma and brain tissues by ultracentrifugation and molecular size exclusion and identified by electron microscopy, particle size analysis, and protein blotting. Long-chain RNA libraries of exosomes were constructed and sequenced. Real-time quantitative reverse transcription polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of inflammatory factors and neutrophil-related chemokines in exosomes from plasma and brain tissues of each group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the protein expression of inflammatory factors and neutrophil-related chemokines in exosomes from brain tissues of each group. Immunohistochemistry was used to detect the expression of the neutrophil-specific protein myeloperoxidase (MPO) in the brains of mice in each group. ResultsCompared with the sham-operated group, the model group showed decreased neurological function scores (P<0.01), obvious cerebral infarction (P<0.01), reduced cerebral blood flow (P<0.01), neuronal necrosis in the brain, and decreased numbers of Nissl bodies (P<0.01). The mRNA expression levels of IL-1β, MPO, CXCL1, CXCL2, CXCL3, CXCL10, CCL2, and CCL3 in exosomes from plasma and brain tissues were significantly increased (P<0.05, P<0.01). The protein expression levels of IL-1β, MPO, CXCL2, and CXCL10 in exosomes from brain tissues were increased (P<0.05, P<0.01), and MPO-positive rates and mean optical density values in brain tissues were elevated (P<0.01). Compared with the model group, the Huanglian Jiedutang group and the Ginaton group showed increased neurological function scores (P<0.05), reduced cerebral infarct volume (P<0.01), restored cerebral blood flow (P<0.01), reduced necrotic cells in the brain, and increased numbers of Nissl bodies (P<0.01). In the Huanglian Jiedutang group, the mRNA expression levels of IL-1β, MPO, CXCL1, CXCL2, CXCL3, CXCL10, CCL2, and CCL3 in exosomes from plasma and brain tissues were decreased (P<0.05, P<0.01). The protein expression levels of IL-1β, MPO, CXCL2, and CXCL10 in exosomes from brain tissues were reduced (P<0.05, P<0.01), and MPO-positive rates and mean optical density values in brain tissues were decreased (P<0.01). ConclusionHuanglian Jiedutang can effectively regulate the expression of neutrophil-related chemokines in exosomes from plasma and brain tissues of MCAO mice, thereby reducing neutrophil infiltration in the brain and achieving therapeutic effects.
7.Tracing Development of LIU Wansu's Theory of ''Fire-heat Inducing Stroke''
Xin LAN ; Changxiang LI ; Haojia ZHANG ; Jialin CHENG ; Zijin SUN ; Liyang DONG ; Zilin REN ; Xueqian WANG ; Fafeng CHENG ; Qingguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):32-41
LIU Wansu, as the foremost of the four great masters of the Jin-Yuan period, established the "theory of fire-heat'' and extended the fire-heat pathogenesis framework to the field of stroke, thereby forming the theory of ''fire-heat inducing stroke''. This achieved a paradigmatic shift in stroke etiology from ''exogenous wind inducing stroke'' to ''fire-heat inducing stroke''. This paper systematically reviews the developmental trajectory of LIU Wansu's ''fire-heat inducing stroke'' theory and explores the social background, academic origins, and core connotations of its theoretical construction. The study found that, based on the ''Nineteen Pathomechanisms'' in the Huangdi's Internal Classic (Huang Di Nei Jing) and combined with clinical practice, LIU Wansu proposed that fire-heat is the fundamental cause of stroke, and that the Six Climatic Factors and the Five Zhi-Emotions can all transform into fire. He further constructed a stratified syndrome differentiation and therapeutic system centered on clearing heat and purging fire, emphasizing differentiated treatment of exterior and interior syndromes, Six Meridians syndrome differentiation, and seasonally adjusted medication. This theory not only resolved the diagnostic and therapeutic dilemmas of febrile epidemic diseases during the Jin-Yuan period, but also exerted a profound influence on later physicians such as ZHANG Zihe and ZHU Danxi, thereby promoting the pluralistic development of stroke theory in traditional Chinese medicine (TCM). Modern pharmacological research provides solid scientific evidence, confirming that the ''fire-heat'' pathological state is highly associated with key mechanisms such as excessive inflammatory responses, oxidative stress, and excitatory amino acid toxicity following cerebral ischemia. Heat-clearing and fire-purging prescriptions and agents, such as Huanglian Jiedu Tang and baicalin, can exert multi-target neuroprotective effects by regulating inflammatory signaling, enhancing antioxidant enzyme activity, and balancing neurotransmitters. This not only verifies the scientific basis of the ''fire-heat inducing stroke'' theory from a modern biological perspective but also provides conclusive evidence for the clinical application of heat-clearing and fire-purging therapy. LIU Wansu's ''fire-heat inducing stroke'' theory represents a major milestone in the historical understanding of stroke pathogenesis, and its academically transitional insights continue to hold core guiding value for the pattern identification and treatment of ischemic stroke today.
8.Effects of Huanglian Jiedutang on Neutrophil Infiltration in Brain of MCAO Mice via Regulation of Chemokine Expression in Exosomes
Haojia ZHANG ; Kai WANG ; Zijin SUN ; Chunyu WANG ; Wei SHAO ; Kunjing LIU ; Liyang DONG ; Dan CHEN ; Wenxiu XU ; Chuanzun WANG ; Wen WANG ; Changxiang LI ; Xueqian WANG ; Fafeng CHENG ; Qingguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):42-53
ObjectiveTo investigate whether Huanglian Jiedutang can inhibit neutrophil infiltration in the brains of middle cerebral artery occlusion (MCAO) mice by regulating the expression of neutrophil-related chemokines in exosomes, thereby achieving therapeutic effects. MethodsA total of 130 male specific pathogen-free (SPF) C57BL/6J mice were randomly divided into four groups: Sham-operated group, MCAO model group, Huanglian Jiedutang group (6 g·kg-1), and Ginaton group (21.6 mg·kg-1), with 10 mice in the Ginaton group and 40 mice in each of the remaining three groups. Mice in the Huanglian Jiedutang group and the Ginaton group were administered the corresponding drugs by oral gavage once daily at a volume of 0.15 mL·(10 g)-1 for 7 consecutive days, while the sham-operated and model groups received an equal volume of saline via the same route. After 7 days, MCAO surgery was performed. The distal and proximal ends of the right common carotid artery (CCA) were ligated, a small incision was made between the two ligatures, and a silicone rubber-coated monofilament with a rounded tip was inserted into the lumen to occlude the CCA. The filament was left in place for 1 h to establish a focal cerebral ischemia model. At 24 h after modeling, mice were evaluated. Neurological function was assessed using the Longa score. Cerebral infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Cerebral blood flow was observed by laser speckle imaging. Hematoxylin and eosin (HE) staining and Nissl staining were used to observe pathological changes in brain tissues. Exosomes were isolated from mouse plasma and brain tissues by ultracentrifugation and molecular size exclusion and identified by electron microscopy, particle size analysis, and protein blotting. Long-chain RNA libraries of exosomes were constructed and sequenced. Real-time quantitative reverse transcription polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of inflammatory factors and neutrophil-related chemokines in exosomes from plasma and brain tissues of each group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the protein expression of inflammatory factors and neutrophil-related chemokines in exosomes from brain tissues of each group. Immunohistochemistry was used to detect the expression of the neutrophil-specific protein myeloperoxidase (MPO) in the brains of mice in each group. ResultsCompared with the sham-operated group, the model group showed decreased neurological function scores (P<0.01), obvious cerebral infarction (P<0.01), reduced cerebral blood flow (P<0.01), neuronal necrosis in the brain, and decreased numbers of Nissl bodies (P<0.01). The mRNA expression levels of IL-1β, MPO, CXCL1, CXCL2, CXCL3, CXCL10, CCL2, and CCL3 in exosomes from plasma and brain tissues were significantly increased (P<0.05, P<0.01). The protein expression levels of IL-1β, MPO, CXCL2, and CXCL10 in exosomes from brain tissues were increased (P<0.05, P<0.01), and MPO-positive rates and mean optical density values in brain tissues were elevated (P<0.01). Compared with the model group, the Huanglian Jiedutang group and the Ginaton group showed increased neurological function scores (P<0.05), reduced cerebral infarct volume (P<0.01), restored cerebral blood flow (P<0.01), reduced necrotic cells in the brain, and increased numbers of Nissl bodies (P<0.01). In the Huanglian Jiedutang group, the mRNA expression levels of IL-1β, MPO, CXCL1, CXCL2, CXCL3, CXCL10, CCL2, and CCL3 in exosomes from plasma and brain tissues were decreased (P<0.05, P<0.01). The protein expression levels of IL-1β, MPO, CXCL2, and CXCL10 in exosomes from brain tissues were reduced (P<0.05, P<0.01), and MPO-positive rates and mean optical density values in brain tissues were decreased (P<0.01). ConclusionHuanglian Jiedutang can effectively regulate the expression of neutrophil-related chemokines in exosomes from plasma and brain tissues of MCAO mice, thereby reducing neutrophil infiltration in the brain and achieving therapeutic effects.
9.Evaluation of the effect of clinical pharmacists participating in the treatment of chronic heart failure based on the clinical pharmacy pathway
Guanhua HOU ; Baozhen WANG ; Yuchen TANG ; Jie CHENG ; Yuan DONG ; Zhiqiang DONG
China Pharmacy 2026;37(6):800-805
OBJECTIVE To evaluate the effect of clinical pharmacists participating in the treatment of chronic heart failure (CHF) based on the clinical pharmacy pathway (CPP). METHODS Totally 226 CHF patients recruited from August 24th, 2024 to March 14th, 2025, were divided into an observation group and a control group based on the random number table method, with 113 cases in each group. All patients were treated with conventional therapy. The observation group was additionally given CPP management (including pharmaceutical care during hospitalization, the formulation of individualized discharge medication regimens, and pharmaceutical follow-up after discharge). The cardiac function parameters at admission, at discharge, at 3 and 6 months after discharge, drug use at 6 months after discharge, economic indicators, as well as the readmission rate and mortality rate at 6 months after discharge were compared between the two groups. Morisky Medication Adherence Scale-8 Items (MMAS-8), Somatic Self-rating Scale (SSS) and Patient Health Questionnaire-9 (PHQ-9) scores were compared at admission, at discharge and at 3 and 6 months after discharge. RESULTS Six months after discharge, 24 patients dropped out. Eventually, 104 patients in the observation group and 98 patients in the control group completed the study. Compared with at admission, New York Heart Association (NYHA) cardiac functional classification, left ventricular ejection fraction (LVEF) and N -terminal pro-B-type natriuretic peptide (NT-proBNP) of both groups of patients at discharge as well as at 3 and 6 months after discharge were significantly improved; moreover, the improvements at 3 and 6 months after discharge were significantly better than those at discharge. Meanwhile, the above indexes (except for NYHA cardiac functional classification at discharge, NT-proBNP and NYHA cardiac functional classification at 3 months after discharge) of the observation group at discharge, at 3 and 6 months after discharge were significantly better than the control group ( P <0.05). The utilization rates of angiotensin converting enzyme inhibitor (ACEI)/angiotensin Ⅱ receptor blocker (ARB)/angiotensin receptor neprilysin inhibitor (ARNI), the proportion of β-blockers reaching the target dose, the utilization rate of sodium-glucose linked transporter 2 inhibitor (SGLT2i), and the proportion of SGLT2i reaching the target dose in the observation group were significantly higher than the control group ( P <0.05), and the proportion of drugs and readmission rate were significantly lower than the control group ( P <0.05). Compared with at admission, MMAS-8 scores of the patients in the observation group at discharge, at 3 and 6 months after discharge were significantly increased, while SSS and PHQ-9 scores were significantly lowered ( P <0.05). And all the above scores gradually decreas ed with the extension of discharge time ( P <0.05). CONCLUSIONS Clinical pharmacists can utilize CPP to significantly improve patients’ cardiac function, medication adherence, somatic symptoms and depression. Additionally, they can significantly improve the utilization rates of ACEI/ARB/ARNI and SGLT2i, as well as the proportion of target doses of β-blockers and SGLT2i, while simultaneously reducing readmission rates.
10.Mechanisms of Huanglian Jiedutang and Its Major Active Constituents in Inhibiting LPS-induced M1 Polarisation of BV2 Microglia
Haojia ZHANG ; Kai WANG ; Kunjing LIU ; Xin LAN ; Zijin SUN ; Chunyu WANG ; Wenyuan MA ; Wei SHAO ; Jinhua HAN ; Liyang DONG ; Changxiang LI ; Xueqian WANG ; Youxiang CUI ; Fafeng CHENG ; Qingguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):44-55
ObjectiveTo investigate whether Huanglian Jiedutang (HLJD) and its major active constituents (geniposide, baicalin, and berberine) can inhibit the inflammatory response of BV2 cells under lipopolysaccharide (LPS) stimulation via the high-mobility group protein B1 (HMGB1)/Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway, and to explore differences in therapeutic efficacy among the three monomers, their combined formula, and HLJD under equal content ratios. MethodsBV2 microglial cells were used as the primary experimental model. Cell viability was assessed using the cell counting kit-8 (CCK-8) method to examine the effects of different concentrations of dimethyl sulfoxide (DMSO, 0.8%, 0.4%, 0.2%, 0.1%, and 0.05%) on cell viability. IncuCyte was employed to monitor the growth of cells under different concentrations of HLJD (200, 100, 50, 25, 12.5, 6.25 mg·L-1). Nitric oxide (NO) assay was used to screen the optimal HLJD concentration. High-performance liquid chromatography (HPLC) determined the content of geniposide, baicalin, and berberine in HLJD, and experimental groups were subsequently established according to the relative proportions of these constituents. CCK-8 assay evaluated cell viability under different treatments. Enzyme-linked immunosorbent assay (ELISA) measured levels of inflammatory factors (TNF-α, IL-1β, IL-6, IL-10) in the supernatant. Flow cytometry assessed the effects of treatments on M1-type polarization of BV2 cells. Western blot determined the expression levels of HMGB1, TLR4, and NF-κB-related proteins. ResultsCompared with the blank group, DMSO at concentrations ≤0.2% did not affect cell viability within 48 h. BV2 cell growth plateaued at 24 h after treatment with 200 mg·L-1 HLJD. Under stimulation with 2 mg·L-1 LPS, this concentration of HLJD effectively reduced NO release, and 6 h pre-treatment had a stronger inhibitory effect on NO than direct administration. HPLC results showed that 1 mg of HLJD freeze-dried powder contained approximately 24 μg of geniposide, 15 μg of baicalin, and 30 μg of berberine. Based on these ratios, experimental groups were blank, LPS (2 mg·L-1), HLJD (200 mg·L-1), monomer combination, geniposide (4.8 mg·L-1), baicalin (3 mg·L-1), and berberine (6 mg·L-1). The monomer combination group consisted of all three active constituents dissolved together. LPS and HLJD or its active constituents did not affect cell viability compared with the blank group. LPS significantly increased TNF-α, IL-1β, IL-6, and IL-10 in the supernatant (P<0.01). HLJD and its active constituents significantly reduced pro-inflammatory factors TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01) while upregulating anti-inflammatory IL-10 (P<0.01), with the monomer combination showing the strongest effect (P<0.05, P<0.01). Compared with the blank group, LPS significantly increased the proportion of CD80⁺CD86⁺ (M1-type) BV2 cells (P<0.01). HLJD and its constituents partially inhibited M1 polarization (P<0.05, P<0.01), with the monomer combination exhibiting the most pronounced effect (P<0.05, P<0.01). Compared with the blank group, LPS upregulated HMGB1, TLR4, and NF-κB-related proteins (P<0.01), whereas HLJD and its active constituents significantly reduced their expression (P<0.05, P<0.01), with the monomer combination having the strongest regulatory effect (P<0.05, P<0.01). ConclusionHLJD and its major active constituents (geniposide, baicalin, berberine) can inhibit LPS-induced inflammatory responses in BV2 cells. The combination of the three active constituents demonstrates the most potent anti-inflammatory effect, significantly attenuating M1-type polarization of BV2 cells via the HMGB1/TLR4/NF-κB signaling pathway.

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