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.

ObjectiveTo investigate the effects of Huanglian Jiedutang （HLJDT） on cognitive function in mice with ischemic stroke （IS） and to elucidate whether its neuroprotective effects are mediated by inhibition of the nuclear factor-κB （NF-κB） signaling pathway and subsequent suppression of NF-κB-regulated neuronal apoptosis. MethodsAn IS model was established using middle cerebral artery occlusion （MCAO）. Sixty C57BL/6J mice were randomly assigned to five groups （n =12 per group）， i.e.， sham operation， model， HLJDT low-dose （3.9 g·kg^-1·d^-1）， HLJDT high-dose （7.8 g·kg^-1·d^-1）， and Ginkgo biloba extract （GBE， 31.2 mg·kg^-1·d^-1）. Post-operatively， neurological deficit scores （Longa score）， cerebral infarct volume assessed by 2，3，5-triphenyltetrazolium chloride （TTC） staining， and brain water content were evaluated. Learning and memory were assessed using new object recognition （NOR） and fear conditioning （FC） tests. Hippocampal pathology was examined via hematoxylin and eosin （HE） staining. Immunofluorescence detected expression of glial fibrillary acidic protein （GFAP， astrocyte marker）， cellular oncogene Fos （c-Fos， neuronal activation marker）， and glutamate decarboxylase 65 （GAD65）. Western blot measured nuclear factor-κB inhibitor protein α （IκBα）， phosphorylated IκBα （p-IκBα）， NF-κB p65， phosphorylated NF-κB p65 （p-NF-κB p65）， ionic calcium binding adapter molecule 1 （Iba-1）， tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and apoptosis-related proteins， such as cleaved cysteinyl aspartate-specific protease 3 （Caspase-3）， B-cell lymphoma 2 （Bcl-2）， and Bcl-2-associated X protein （Bax）. Real-time quantitative PCR （Real-time PCR） was used to assess mRNA levels of Iba-1， TNF-α， IL-1β， NF-κB p65， cleaved Caspase-3， Bax， and Bcl-2. ResultsCompared with the sham group， the model group exhibited significantly increased neurological deficit scores， brain water content， and cerebral infarct volume （P<0.01）. Hippocampal CA1 neurons were disorganized， showing nuclear pyknosis and karyolysis. NOR exploration time and FC freezing time were significantly reduced （P<0.01）. GFAP and c-Fos expression were increased， while GAD65 expression was decreased （P<0.01）. Cleaved Caspase-3 and Bax were upregulated， Bcl-2 was downregulated， and the Bax/Bcl-2 ratio was elevated （P<0.01）. Expression levels of p-IκBα， p-NF-κB p65， IL-1β， TNF-α， and Iba-1 were significantly increased （P<0.01）. Compared with the model group， HLJDT high-dose， low-dose， and GBE groups showed significant improvements in all parameters （P<0.01）. Among them， the HLJDT high-dose group showed the most pronounced neuronal structural recovery and superior performance in NOR and FC tests （P<0.01）. In this group， GFAP and c-Fos decreased， GAD65 increased （P<0.01）， apoptosis-related protein expression was reversed， and NF-κB signaling and related inflammatory factor expression were suppressed （P<0.01）. ConclusionHLJDT ameliorates cognitive dysfunction in mice after IS， potentially by inhibiting the NF-κB signaling pathway， thereby reducing neuroinflammation and hippocampal neuronal apoptosis.

Huanglian Jiedutang （HLJDT）， as a classical formula for clearing heat and removing toxins， has been widely applied in the treatment of various clinical diseases in recent years， particularly during the fire-heat stage of stroke， where it has attracted considerable attention. Based on previous studies， this paper systematically elaborates on the research progress on the active components of HLJDT， its clinical application in ischemic stroke， and advances in studies on its mechanisms of action. Modern pharmacological studies have demonstrated that HLJDT contains multiple active components， including baicalin， geniposide， and berberine. In the treatment of ischemic stroke， these components exert therapeutic effects through multi-target， multi-pathway， and multi-level mechanisms. Clinical studies have shown that HLJDT can increase cerebral blood flow， reduce cerebral infarct volume， and improve post-stroke physical dysfunction in patients with ischemic stroke. Experimental studies have indicated that HLJDT can improve neurological function scores and increase cerebral perfusion in experimental stroke models. In addition， the mechanisms underlying the anti-ischemic stroke effects of HLJDT may be related to anti-inflammatory and antioxidant activities， promotion of angiogenesis， and regulation of amino acid and energy metabolism. Although existing studies have confirmed that HLJDT exhibits multi-target and multi-pathway synergistic therapeutic characteristics， further large-sample randomized controlled trials are still needed to verify its long-term efficacy and to further elucidate the dynamic interaction network among components， targets， and pathways. Combined with network pharmacology and molecular docking analyses， this study further clarifies the synergistic targets of the core components （berberine， baicalin， and geniposide）， providing a theoretical basis for in-depth research and clinical translation of HLJDT in the treatment of ischemic stroke.

Background: and Purpose Ruptured intracranial aneurysms (RIA) are associated with a mortality rate of up to 40% in the Chinese population, highlighting the critical need for targeted treatment interventions for at-risk individuals. Although the impact of aldehyde dehydrogenase 2 (ALDH2) gene mutations on susceptibility to intracranial aneurysms (IA) is well documented, the potential connection between ALDH2 rs671 single-nucleotide polymorphism (SNP) and RIA remains unexplored. Given the increased prevalence of ALDH2 gene mutations among Chinese Han individuals, it is clinically relevant to investigate the link between ALDH2 rs671 SNP and IA rupture. Methods: A prospective study was conducted on 546 patients diagnosed with IA to investigate the association between ALDH2 rs671 SNP and the risk of IA rupture. Results: The ALDH2 rs671 SNP (ALDH2*2) was significantly more prevalent in patients with unruptured IA (UIA) than in those with RIA (32.56% vs. 18.58%, P=0.004). Multivariate logistic regression analysis revealed that people with the ALDH2 mutation (ALDH2*1/*2 and ALDH2*2/*2 gene type) had a significantly reduced odds ratio (OR=0.49; 95% confidence level [CI] 0.27–0.88; P=0.018) for RIAs. Age-specific subgroup analysis indicated that the ALDH2 mutation provided a stronger protective effect in individuals aged 60 years and above with IA compared to those under 60 years old (OR=0.38 vs. OR=0.52, both P<0.05). Conclusion The incidence of RIA was significantly higher in individuals with a normal ALDH2 gene (ALDH2*1/*1) than in those with an ALDH2 rs671 SNP (ALDH2*1/*2 or ALDH2*2/*2). ALDH2 rs671 SNP may serve as a protective factor against RIA in the Chinese Han population.

Background: and Purpose Ruptured intracranial aneurysms (RIA) are associated with a mortality rate of up to 40% in the Chinese population, highlighting the critical need for targeted treatment interventions for at-risk individuals. Although the impact of aldehyde dehydrogenase 2 (ALDH2) gene mutations on susceptibility to intracranial aneurysms (IA) is well documented, the potential connection between ALDH2 rs671 single-nucleotide polymorphism (SNP) and RIA remains unexplored. Given the increased prevalence of ALDH2 gene mutations among Chinese Han individuals, it is clinically relevant to investigate the link between ALDH2 rs671 SNP and IA rupture. Methods: A prospective study was conducted on 546 patients diagnosed with IA to investigate the association between ALDH2 rs671 SNP and the risk of IA rupture. Results: The ALDH2 rs671 SNP (ALDH2*2) was significantly more prevalent in patients with unruptured IA (UIA) than in those with RIA (32.56% vs. 18.58%, P=0.004). Multivariate logistic regression analysis revealed that people with the ALDH2 mutation (ALDH2*1/*2 and ALDH2*2/*2 gene type) had a significantly reduced odds ratio (OR=0.49; 95% confidence level [CI] 0.27–0.88; P=0.018) for RIAs. Age-specific subgroup analysis indicated that the ALDH2 mutation provided a stronger protective effect in individuals aged 60 years and above with IA compared to those under 60 years old (OR=0.38 vs. OR=0.52, both P<0.05). Conclusion The incidence of RIA was significantly higher in individuals with a normal ALDH2 gene (ALDH2*1/*1) than in those with an ALDH2 rs671 SNP (ALDH2*1/*2 or ALDH2*2/*2). ALDH2 rs671 SNP may serve as a protective factor against RIA in the Chinese Han population.

Objective:To evaluate the association between triglyceride-glucose(TyG) index level, their variability, and the risk of incident hyperuricemia(HUA).Methods:A total of 1 583 cases with good compliance who underwent follow-up at the health examination center of a tertiary hospital physical in Zhengzhou were enrolled. The TyG index mean(TyG-mean) and variability indexes, including standard deviation(TyG-SD), coefficient of variation(TyG-CV), and adjusted standard deviation(adj-TyG-SD), were calculated based on TyG index values from three consecutive annual health check-ups. Cox proportional risk regression model was used to assess the relationship between the variability of TyG index and the risk of new-onset HUA; the dose-response relationship between different TyG indexes and HUA was examined using restricted cubic spline(RCS). Results:After a 3-year follow-up, 146 participants developed incident HUA. Both TyG-mean and TyG index variability indicators were significantly higher in the HUA group compared to the non-HUA group( P<0.05). After adjusting for multiple confounders, each standard deviation σincrease in TyG-SD, TyG-CV, and adj-TyG-SD was associated with a 1.23-fold(95% CI 1.06-1.43), 1.22-fold(95% CI 1.05-1.42), and 1.26-fold(95% CI 1.08-1.45) higher risk of incident HUA, respectively. RCS analysis revealed a nonlinear association between adj-TyG-SD and HUA risk( P<0.05), with a critical threshold of 0.55 at a hazard ratio( HR) of 1. Conclusions:Increased TyG index variability is associated with a higher risk of incident HUA, with adj-TyG-SD showing the strongest correlation with HUA risk.

Background: and Purpose Ruptured intracranial aneurysms (RIA) are associated with a mortality rate of up to 40% in the Chinese population, highlighting the critical need for targeted treatment interventions for at-risk individuals. Although the impact of aldehyde dehydrogenase 2 (ALDH2) gene mutations on susceptibility to intracranial aneurysms (IA) is well documented, the potential connection between ALDH2 rs671 single-nucleotide polymorphism (SNP) and RIA remains unexplored. Given the increased prevalence of ALDH2 gene mutations among Chinese Han individuals, it is clinically relevant to investigate the link between ALDH2 rs671 SNP and IA rupture. Methods: A prospective study was conducted on 546 patients diagnosed with IA to investigate the association between ALDH2 rs671 SNP and the risk of IA rupture. Results: The ALDH2 rs671 SNP (ALDH2*2) was significantly more prevalent in patients with unruptured IA (UIA) than in those with RIA (32.56% vs. 18.58%, P=0.004). Multivariate logistic regression analysis revealed that people with the ALDH2 mutation (ALDH2*1/*2 and ALDH2*2/*2 gene type) had a significantly reduced odds ratio (OR=0.49; 95% confidence level [CI] 0.27–0.88; P=0.018) for RIAs. Age-specific subgroup analysis indicated that the ALDH2 mutation provided a stronger protective effect in individuals aged 60 years and above with IA compared to those under 60 years old (OR=0.38 vs. OR=0.52, both P<0.05). Conclusion The incidence of RIA was significantly higher in individuals with a normal ALDH2 gene (ALDH2*1/*1) than in those with an ALDH2 rs671 SNP (ALDH2*1/*2 or ALDH2*2/*2). ALDH2 rs671 SNP may serve as a protective factor against RIA in the Chinese Han population.

Objective:To evaluate the association between triglyceride-glucose(TyG) index level, their variability, and the risk of incident hyperuricemia(HUA).Methods:A total of 1 583 cases with good compliance who underwent follow-up at the health examination center of a tertiary hospital physical in Zhengzhou were enrolled. The TyG index mean(TyG-mean) and variability indexes, including standard deviation(TyG-SD), coefficient of variation(TyG-CV), and adjusted standard deviation(adj-TyG-SD), were calculated based on TyG index values from three consecutive annual health check-ups. Cox proportional risk regression model was used to assess the relationship between the variability of TyG index and the risk of new-onset HUA; the dose-response relationship between different TyG indexes and HUA was examined using restricted cubic spline(RCS). Results:After a 3-year follow-up, 146 participants developed incident HUA. Both TyG-mean and TyG index variability indicators were significantly higher in the HUA group compared to the non-HUA group( P<0.05). After adjusting for multiple confounders, each standard deviation σincrease in TyG-SD, TyG-CV, and adj-TyG-SD was associated with a 1.23-fold(95% CI 1.06-1.43), 1.22-fold(95% CI 1.05-1.42), and 1.26-fold(95% CI 1.08-1.45) higher risk of incident HUA, respectively. RCS analysis revealed a nonlinear association between adj-TyG-SD and HUA risk( P<0.05), with a critical threshold of 0.55 at a hazard ratio( HR) of 1. Conclusions:Increased TyG index variability is associated with a higher risk of incident HUA, with adj-TyG-SD showing the strongest correlation with HUA risk.

Objective:To explore the potential correlation between cumulative pulse pressure (cumPP) level and metabolic syndrome (MetS), and to provide insights for MetS management.Methods:A total of 3 968 subjects who underwent health checkup were selected to form a research cohort, and the data were categorized into three groups based on the tertiles of cumPP levels. Cox proportional hazards regression model was employed to analyze the association between different cumPP levels and the incidence of new-onset MetS. Results:The risk of MetS increased with the increased tiers of the cumPP levels (2.5%, 4.3%, and 4.6%, Ptrend<0.001) during the median follow-up period of 2.16 years. Spearman rank correlation analysis showed that cumPP was positively correlated with waist circumference, systolic blood pressure, diastolic blood pressure and fasting plasma glucose (all P<0.05). The Cox proportional hazards regression adjusted model showed that the risk of MetS in Q2 and Q3 was higher than that in Q1 in the total population, and the same results were observed in males (all P<0.05), while there was no statistical significance in females. Model 3 of the total population adjusted for a variety of confounding factors displayed a higher risk of MetS in Q3 compared with that in Q1[1.654 (95% CI 1.272-2.151) ]. When stratified by sex, and the risk of MetS in Q3 was 1.665 times higher than that in Q1 (95% CI 1.245-2.227), while there was no statistically significant risk in female. According to the visual nomogram of independent risk factors screened by multivariate analysis based on Cox proportional hazards regression model, the incidence of MetS at 1 year, 2 years, and 3 years was 0.18%, 3.97% and 7.39%, respectively. In addition, the dose-response curve was plotted according to cumPP, suggesting that the risk of MetS gradually increased with the increase of cumPP in the total population. Subgroup analyses based on baseline systolic blood pressure levels showed that higher cumPP levels were associated with a higher risk of developing MetS, regardless of whether systolic blood pressure was abnormal. Conclusions:Elevated cumPP levels is significantly related to the incidence of new-onset MetS. Maintaining pulse pressure within an appropriate range over long term is crucial for the management of MetS.

Objective:To explore the relationship between the long-term dynamic change in alanine aminotransferase(ALT) level and metabolic associated fatty liver disease(MAFLD).Methods:A retrospective study was conducted on 6 864 subjects who underwent four consecutive physical examinations from 2017 to 2020 in a cohort study of physical examination population in Henan Province. The relation between ALT level and the shift of MAFLD risk was analyzed using a multi-state Markov model, and the bidirectional relationship between ALT level and MAFLD was explored using a random intercept cross-lagged model.Results:Multi-state Markov model after adjusting for confounding factors showed that the risk of MAFLD in ALT Q2, Q3, Q4 group was gradually higher than that in Q1 group; Compared with health status, non-alcoholic fatty liver disease and MAFLD status gradually increased the risk of ALT shifting from normal to abnormal. The random intercept cross-lagged model after adjusting for confounding factors showed that there was a significant positive bidirectional relationship between MAFLD and ALT level. The cross-lag effect of MAFLD→ALT level was 0.083(95% CI 0.078-0.087), and the cross-lag effect of ALT→MAFLD was 0.044(95% CI 0.039-0.050). And with the extension of time, the cross-lag effect gradually decreased. Conclusions:There is a significant bidirectional relationship between the long-term dynamic change of ALT level and MAFLD. The occurrence of MAFLD is more likely to increase the risk of elevated ALT level, emphasizing the need for enhanced early prevention and treatment of MAFLD.