1.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.
2.Banxia Baizhu Tianmatang Regulates NLRP3 Inflammasomes to Ameliorate Cognitive Impairment in Epilepsy
Xingdan ZHU ; Yinhua KAI ; Rong TIAN ; Xin YANG ; Jiayi HE ; Xiangxin GUO ; Yadong MU ; Cui JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):308-316
ObjectiveTo explore the mechanism through which Banxia Baizhu Tianmatang ameliorates cognitive impairment in epileptic rats induced by lithium chloride-pilocarpine by regulating the neuroinflammatory reaction mediated by NOD-like receptor protein 3 (NLRP3) inflammasomes. MethodsSixty male SD rats were randomly allocated into blank, model, carbamazepine (0.125 g·kg-1·d-1), Banxia Baizhu Tianmatang (1.04 g·kg-1·d-1), and carbamazepine (0.125 g·kg-1·d-1) + Banxia Baizhu Tianmatang (1.04 g·kg-1·d-1) groups (n=12). After the modeling of epilepsy, rats were administrated with corresponding agents by gavage for 12 weeks. At the 6th and 12th week of the intervention, the rats’ hyper-excited behavior was evaluated by the stylus experiment, and at the 12th week of intervention, the cognitive function was evaluated by Barnes maze. At the same time, the seizure frequency and severity grade (Racine score) were recorded. The serum and hippocampus tissue samples were collected after anesthesia for the following tests. Nissl staining was used to evaluate the degree of neuronal damage in the hippocampal CA1 area. The content of malondialdehyde (MDA) in the hippocampus was determined by the thiobarbituric acid (TBA) method. Serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18) were quantified by enzyme-linked immunosorbent assay (ELISA). Immunohistochemical method was adopted to detect the expression of apoptosis-associated speck-like protein containing a card (ASC) in the hippocampus. Western blot was employed to quantitatively analyze the protein levels of NLRP3, cysteinyl aspartate-specific proteinase-1 (Caspase-1), and brain-derived neurotrophic factor (BDNF) in the hippocampus. ResultsThe model group showed increased stylus scores at the 6th and 12th week after modeling, a decreased Barnes maze strategy score at the 12th week, a prolonged incubation period (P<0.05), elevated serum levels of inflammatory factors (P<0.05), decreased neurons with scattered arrangement and large gaps in the hippocampus, increased content of MDA in the hippocampus (P<0.05), an increased positive expression of ASC, and up-regulated protein levels of Caspase-1, NLRP3, and BDNF (P<0.05). Compared with the model group, the intervention with Banxia Baizhu Tianmatang for 12 weeks was accompanied by a decreased stylus score, epileptic seizures with a decreased score, a decreased number, and shortened duration, an increased Barnes maze strategy score, shortened escape latency (P<0.01), declined serum levels of inflammatory factors (P<0.05), regular morphology of hippocampal neurons, reduced MDA content in the hippocampus (P<0.05), a decreased positive expression of ASC, and down-regulated protein levels of Caspase-1, NLRP3, and BDNF (P<0.05, P<0.01). In addition, compared with the carbamazepine group, Banxia Baizhu Tianmatang + carbamazepine showed improved performance in controlling the seizure, improved the cognitive behavior score and morphology of hippocampal neurons, alleviated the oxidative stress products, lowered the levels of inflammatory factors, reduced the positive expression of ASC in the hippocampus, and down-regulated the expression of Caspase-1, NLRP3 and BDNF, with no significant differences. ConclusionBanxia Baizhu Tianmatang may reduce neuroinflammation, control epileptic seizures, and ameliorate cognitive impairment by inhibiting the expression of NLRP3 inflammasomes.
3.Exploring Mechanism of Banxia Baizhu Tianma Decoction in the Treatment of Epilepsy Based on Network Pharmacology and Experimental Verification
Xin YANG ; Jin FU ; Cui JIANG ; Yinhua KAI ; Jiayi HE ; Xiangxin GUO ; Rong TIAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(3):776-791
Objective To analyze the target,signal pathway and potential mechanism of Banxia Baizhu Tianma Decoction in the treatment of epilepsy based on network pharmacology,and to verify it by molecular docking technology and animal experiments.Methods The active ingredients and drug targets of Banxia Baizhu Tianma Decoction were screened by BATMAN and other databases.The targets of epilepsy-related diseases were obtained by GeneCards and other databases,and the intersection targets were taken.Constructing'drug-ingredient-target-disease'network and PPI network to screen the core targets.The core active ingredients were screened according to GO,KEGG functional enrichment analysis and'pathway-target-active ingredient'network.Molecular docking was used to verify the core targets and core active ingredients.In the animal experiment,the rat model of epilepsy was induced by lithium chloride-pilocarpine,and 40 Wistar rats were divided into normal control group,model control group,carbamazepine group and Banxia Baizhu Tianma Decoction group.The seizures were observed by behavior.Nissl staining was used to observe neuronal damage in hippocampus.Immunohistochemistry was used to detect the expression of BAX,BCL-2 and Caspase-3 protein.Results A total of 1072 targets of Banxia Baizhu Tianma Decoction were screened,1046 disease targets of epilepsy were screened,and 220 intersection targets of Banxia Baizhu Tianma Decoction in the treatment of epilepsy were screened.The core targets AKT1,ALB,ACTB,INS and TNF of PPI network were obtained.KEGG pathway mainly involves TNF signaling pathway,IL-17 signaling pathway,cAMP signaling pathway,pathways of neurodegeneration-multiple diseases,serotonergic synapse and Dopaminergic synapse.The core active ingredients with the highest correlation in the'pathway-target-active component'network were Betulin,Ephedrine,Ergotamine and Thymol.Results of molecular docking indicated that the core target had satisfactory affinity with those active ingredients.Animal experiments showed that Banxia Baizhu Tianma Decoction could effectively reduce epileptic seizures in rats,improve hippocampal neuronal damage in rats,significantly reduce the percentage of neuronal apoptosis,significantly down-regulate the expression of pro-apoptotic proteins BAX and Caspase-3,and up-regulate the expression of anti-apoptotic protein BCL-2.Conclusion Banxia Baizhu Tianma Decoction has the characteristics of multi-component,multi-target and multi-pathway in the treatment of epilepsy.Inhibiting neuronal apoptosis and reducing hippocampal neuronal damage may be one of the important mechanisms for its treatment of epilepsy.
4.Exploring Mechanism of Banxia Baizhu Tianma Decoction in the Treatment of Epilepsy Based on Network Pharmacology and Experimental Verification
Xin YANG ; Jin FU ; Cui JIANG ; Yinhua KAI ; Jiayi HE ; Xiangxin GUO ; Rong TIAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(3):776-791
Objective To analyze the target,signal pathway and potential mechanism of Banxia Baizhu Tianma Decoction in the treatment of epilepsy based on network pharmacology,and to verify it by molecular docking technology and animal experiments.Methods The active ingredients and drug targets of Banxia Baizhu Tianma Decoction were screened by BATMAN and other databases.The targets of epilepsy-related diseases were obtained by GeneCards and other databases,and the intersection targets were taken.Constructing'drug-ingredient-target-disease'network and PPI network to screen the core targets.The core active ingredients were screened according to GO,KEGG functional enrichment analysis and'pathway-target-active ingredient'network.Molecular docking was used to verify the core targets and core active ingredients.In the animal experiment,the rat model of epilepsy was induced by lithium chloride-pilocarpine,and 40 Wistar rats were divided into normal control group,model control group,carbamazepine group and Banxia Baizhu Tianma Decoction group.The seizures were observed by behavior.Nissl staining was used to observe neuronal damage in hippocampus.Immunohistochemistry was used to detect the expression of BAX,BCL-2 and Caspase-3 protein.Results A total of 1072 targets of Banxia Baizhu Tianma Decoction were screened,1046 disease targets of epilepsy were screened,and 220 intersection targets of Banxia Baizhu Tianma Decoction in the treatment of epilepsy were screened.The core targets AKT1,ALB,ACTB,INS and TNF of PPI network were obtained.KEGG pathway mainly involves TNF signaling pathway,IL-17 signaling pathway,cAMP signaling pathway,pathways of neurodegeneration-multiple diseases,serotonergic synapse and Dopaminergic synapse.The core active ingredients with the highest correlation in the'pathway-target-active component'network were Betulin,Ephedrine,Ergotamine and Thymol.Results of molecular docking indicated that the core target had satisfactory affinity with those active ingredients.Animal experiments showed that Banxia Baizhu Tianma Decoction could effectively reduce epileptic seizures in rats,improve hippocampal neuronal damage in rats,significantly reduce the percentage of neuronal apoptosis,significantly down-regulate the expression of pro-apoptotic proteins BAX and Caspase-3,and up-regulate the expression of anti-apoptotic protein BCL-2.Conclusion Banxia Baizhu Tianma Decoction has the characteristics of multi-component,multi-target and multi-pathway in the treatment of epilepsy.Inhibiting neuronal apoptosis and reducing hippocampal neuronal damage may be one of the important mechanisms for its treatment of epilepsy.
5.The evaluation value of CTP and CTA combined with serum NSE in collateral circulation status and prognosis of patients with post-stroke hemiplegia
Xin LIU ; Yu MA ; Kai LU ; Guicheng ZHANG ; Yue WU ; Fangmei HU ; Youxiang CUI ; Yunchuan SUN
Tianjin Medical Journal 2025;53(6):629-633
Objective To investigate the value of CT perfusion imaging(CTP)and CT angiography(CTA)combined with serum neuron-specific enolase(NSE)in assessing the status and prognosis of collateral circulation(CC)in hemiplegic patients with stroke.Methods A total of 106 patients with stroke hemiplegia were selected in this study.All patients underwent CTA and CTP,and patients were classified into the good CC group(n=67)and the poor CC group(n=39)based on CTA images.Patients were also classified into the good prognostic group(n=56)and the poor prognostic group(n=50)based on modified Rankin Scale(mRS)scores after 3 months of treatment.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum NSE levels.Pearson correlation analysis was used to analyse the correlation between CC scores and CTP parameters and serum NSE levels.The receiver operating characteristic(ROC)curve was used to analyze the evaluation value of serum NSE for the prognosis of hemiplegic patients after stroke.Kappa test was used to analyse the consistency of the prognosis and follow-up results of hemiplegic stroke patients assessed by CTP and CTA alone and in combination with serum NSE.Results The CTP parameters rCBF and rCBV were lower in the poor CC group than those in the good CC group(P<0.05),and levels of rTTP,rMTT and serum NSE were higher than those in the good CC group(P<0.01).CC score was positively correlated with rCBF and rCBV,and negatively correlated with rTTP,rMTT and serum NSE levels(P<0.05).The rCBF and rCBV were lower in the poor prognosis group than those in the good prognosis group,and the proportion of poor CC,rTTP,rMTT,serum NSE level and mRS score were higher than those in the good prognosis group(P<0.01).The area under the curve of serum NSE alone for predicting poor prognosis in hemiplegic patients with stroke was 0.878(95%CI:0.800-0.934),with a sensitivity of 74.00%and a specificity of 91.07%,which was in good agreement with the results of the follow-up(Kappa value=0.654,P<0.001).Conclusion CTP,CTA combined with serum NSE have a relatively high evaluation value for the assessment of the CC status and prognosis of stroke patients with hemiplegia.
6.The evaluation value of CTP and CTA combined with serum NSE in collateral circulation status and prognosis of patients with post-stroke hemiplegia
Xin LIU ; Yu MA ; Kai LU ; Guicheng ZHANG ; Yue WU ; Fangmei HU ; Youxiang CUI ; Yunchuan SUN
Tianjin Medical Journal 2025;53(6):629-633
Objective To investigate the value of CT perfusion imaging(CTP)and CT angiography(CTA)combined with serum neuron-specific enolase(NSE)in assessing the status and prognosis of collateral circulation(CC)in hemiplegic patients with stroke.Methods A total of 106 patients with stroke hemiplegia were selected in this study.All patients underwent CTA and CTP,and patients were classified into the good CC group(n=67)and the poor CC group(n=39)based on CTA images.Patients were also classified into the good prognostic group(n=56)and the poor prognostic group(n=50)based on modified Rankin Scale(mRS)scores after 3 months of treatment.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum NSE levels.Pearson correlation analysis was used to analyse the correlation between CC scores and CTP parameters and serum NSE levels.The receiver operating characteristic(ROC)curve was used to analyze the evaluation value of serum NSE for the prognosis of hemiplegic patients after stroke.Kappa test was used to analyse the consistency of the prognosis and follow-up results of hemiplegic stroke patients assessed by CTP and CTA alone and in combination with serum NSE.Results The CTP parameters rCBF and rCBV were lower in the poor CC group than those in the good CC group(P<0.05),and levels of rTTP,rMTT and serum NSE were higher than those in the good CC group(P<0.01).CC score was positively correlated with rCBF and rCBV,and negatively correlated with rTTP,rMTT and serum NSE levels(P<0.05).The rCBF and rCBV were lower in the poor prognosis group than those in the good prognosis group,and the proportion of poor CC,rTTP,rMTT,serum NSE level and mRS score were higher than those in the good prognosis group(P<0.01).The area under the curve of serum NSE alone for predicting poor prognosis in hemiplegic patients with stroke was 0.878(95%CI:0.800-0.934),with a sensitivity of 74.00%and a specificity of 91.07%,which was in good agreement with the results of the follow-up(Kappa value=0.654,P<0.001).Conclusion CTP,CTA combined with serum NSE have a relatively high evaluation value for the assessment of the CC status and prognosis of stroke patients with hemiplegia.
7.Research on Influencing Factors of Satisfaction of Public Emergency Medical Rescue Resources'Demand under the Flood Disaster
Kai CUI ; Peng ZHANG ; Xin ZHANG
Chinese Hospital Management 2024;44(8):93-96
Objective To explore public emergency medical rescue resources'satisfaction of demand,influencing factors and multi-subject enabling countermeasures under the flood disaster.Methods A total of 651 valid question-naires were collected from victims in four towns in one city.Logistic regression analysis was applied to analyze the in-fluencing factors.Results 80.49%of the respondents considered that their and their families'emergency medical res-cue resources'demand could be met during the flood.The results of logistic regression analysis showed influencing factors of low satisfaction including there were the elderly aged 65 and above in the family,and patients with chronic diseases such as diabetes and hypertension in the family and the differences were statistically significant(P<0.05);Influencing factors of high satisfaction including adequate preparation of familial emergency supplies,the grassroots government publicized the disaster prevention and self-help knowledge before the disaster,the grass-roots government had prepared emergency medical rescue resources,such as food,water and relief materials be-fore the disaster,the adequacy of emergency medical rescue resources of local hospital,and medical staff pro-moted psychological knowledge and skill and the differences were statistically significant(P<0.05).Conclusion The store of emergency resource should be strengthened in all aspects,including medical institutions,the grass-roots governments and the public in the community,and emergency preparedness and publicity work should be promoted in a coordinated manner,so as to build a resilient community and society for disaster response.
8.Using machine learning to construct the diagnosis model of female bladder outlet obstruction based on urodynamic study data
Quan ZHOU ; Guang LI ; Kai CUI ; Weilin MAO ; Dongxu LIN ; Zhenglong YANG ; Zhong CHEN ; Youmin HU ; Xin ZHANG
Investigative and Clinical Urology 2024;65(6):559-566
Purpose:
To intelligently diagnose whether there is bladder outlet obstruction (BOO) in female with decent detrusor contraction ability by focusing on urodynamic study (UDS) data.
Materials and Methods:
We retrospectively reviewed the UDS data of female patients during urination. Eleven easily accessible urinary flow indicators were calculated according to the UDS data of each patient during voiding period. Eight diagnosis models based on back propagation neural network with different input feature combination were constructed by analyzing the correlations between indicators and lower urinary tract dysfunction labels. Subsequently, the stability of diagnostic models was evaluated by five-fold cross-validation based on training data, while the performance was compared on test dataset.
Results:
UDS data from 134 female patients with a median age of 51 years (range, 27–78 years) were selected for our study.Among them, 66 patients suffered BOO and the remaining were normal. Applying the 5-fold cross-validation method, the model with the best performance achieved an area under the receiver operating characteristic curve (AUC) value of 0.949±0.060 using 9 UDS input features. The accuracy, sensitivity, and specificity for BOO diagnosis model in the testing process are 94.4%, 100%, and 89.3%, respectively.
Conclusions
The 9 significant indicators in UDS were employed to construct a diagnostic model of female BOO based on machine learning algorithm, which performs preferable classification accuracy and stability.
9.Research on Influencing Factors of Satisfaction of Public Emergency Medical Rescue Resources'Demand under the Flood Disaster
Kai CUI ; Peng ZHANG ; Xin ZHANG
Chinese Hospital Management 2024;44(8):93-96
Objective To explore public emergency medical rescue resources'satisfaction of demand,influencing factors and multi-subject enabling countermeasures under the flood disaster.Methods A total of 651 valid question-naires were collected from victims in four towns in one city.Logistic regression analysis was applied to analyze the in-fluencing factors.Results 80.49%of the respondents considered that their and their families'emergency medical res-cue resources'demand could be met during the flood.The results of logistic regression analysis showed influencing factors of low satisfaction including there were the elderly aged 65 and above in the family,and patients with chronic diseases such as diabetes and hypertension in the family and the differences were statistically significant(P<0.05);Influencing factors of high satisfaction including adequate preparation of familial emergency supplies,the grassroots government publicized the disaster prevention and self-help knowledge before the disaster,the grass-roots government had prepared emergency medical rescue resources,such as food,water and relief materials be-fore the disaster,the adequacy of emergency medical rescue resources of local hospital,and medical staff pro-moted psychological knowledge and skill and the differences were statistically significant(P<0.05).Conclusion The store of emergency resource should be strengthened in all aspects,including medical institutions,the grass-roots governments and the public in the community,and emergency preparedness and publicity work should be promoted in a coordinated manner,so as to build a resilient community and society for disaster response.
10.Research on Influencing Factors of Satisfaction of Public Emergency Medical Rescue Resources'Demand under the Flood Disaster
Kai CUI ; Peng ZHANG ; Xin ZHANG
Chinese Hospital Management 2024;44(8):93-96
Objective To explore public emergency medical rescue resources'satisfaction of demand,influencing factors and multi-subject enabling countermeasures under the flood disaster.Methods A total of 651 valid question-naires were collected from victims in four towns in one city.Logistic regression analysis was applied to analyze the in-fluencing factors.Results 80.49%of the respondents considered that their and their families'emergency medical res-cue resources'demand could be met during the flood.The results of logistic regression analysis showed influencing factors of low satisfaction including there were the elderly aged 65 and above in the family,and patients with chronic diseases such as diabetes and hypertension in the family and the differences were statistically significant(P<0.05);Influencing factors of high satisfaction including adequate preparation of familial emergency supplies,the grassroots government publicized the disaster prevention and self-help knowledge before the disaster,the grass-roots government had prepared emergency medical rescue resources,such as food,water and relief materials be-fore the disaster,the adequacy of emergency medical rescue resources of local hospital,and medical staff pro-moted psychological knowledge and skill and the differences were statistically significant(P<0.05).Conclusion The store of emergency resource should be strengthened in all aspects,including medical institutions,the grass-roots governments and the public in the community,and emergency preparedness and publicity work should be promoted in a coordinated manner,so as to build a resilient community and society for disaster response.

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