1.Correlation Analysis of Huanglian Jiedu Wan on Syndrome Improvement and Clinical Biomarkers of "Excess Heat-Toxicity" Based on Machine Learning Model
Qi LI ; Keke LUO ; Baolin BIAN ; Hongyu YU ; Mengxiao WANG ; Mengyao TIAN ; Wen XIA ; Yuan MA ; Xinfang ZHANG ; Pengyue LI ; Nan SI ; Hongjie WANG ; Yanyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):162-173
ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally, the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome, such as oral ulcers, sore throat, and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation, oxidative stress, and energy metabolism in the early phase, Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting (XGBoost) algorithm was used to develop a prediction model for main symptom classification, with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers, sore throat, and overall symptoms, with significant effects observed especially in sore throat and overall symptom analyses (P<0.01). Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement, were also called "heat-related biomarkers", including succinic acid, α-ketoglutaric acid, glycine, lactic acid, adenosine monophosphate (AMP), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and so on. Conversely, clinical biomarkers negatively correlated with symptom severity, were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan, including malic acid, fumaric acid, cis-aconitic acid, adrenocorticotropic hormone (ACTH), IL-1β, IL-4, IL-8, succinic acid, and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables, with importance scores of 0.081 and 0.080, respectively. After screening out 14 key variables and optimizing the parameters, model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables, confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established, achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.
2.Correlation Analysis of Huanglian Jiedu Wan on Syndrome Improvement and Clinical Biomarkers of "Excess Heat-Toxicity" Based on Machine Learning Model
Qi LI ; Keke LUO ; Baolin BIAN ; Hongyu YU ; Mengxiao WANG ; Mengyao TIAN ; Wen XIA ; Yuan MA ; Xinfang ZHANG ; Pengyue LI ; Nan SI ; Hongjie WANG ; Yanyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):162-173
ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally, the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome, such as oral ulcers, sore throat, and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation, oxidative stress, and energy metabolism in the early phase, Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting (XGBoost) algorithm was used to develop a prediction model for main symptom classification, with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers, sore throat, and overall symptoms, with significant effects observed especially in sore throat and overall symptom analyses (P<0.01). Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement, were also called "heat-related biomarkers", including succinic acid, α-ketoglutaric acid, glycine, lactic acid, adenosine monophosphate (AMP), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and so on. Conversely, clinical biomarkers negatively correlated with symptom severity, were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan, including malic acid, fumaric acid, cis-aconitic acid, adrenocorticotropic hormone (ACTH), IL-1β, IL-4, IL-8, succinic acid, and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables, with importance scores of 0.081 and 0.080, respectively. After screening out 14 key variables and optimizing the parameters, model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables, confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established, achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.
3.The study of m6A methylation-related proteins in the prefrontal cortex of PTSD mice
Jiaying LU ; Luodong YANG ; Keke LU ; Wenlong XIN ; Bin LI ; Qulong LI ; Guiqing ZHANG
Acta Universitatis Medicinalis Anhui 2026;61(3):495-500
ObjectiveTo investigate the expression of prefrontal cortical neurons, methyltransferase-like 3 (METTL3), fat mass and obesity-associated gene (FTO), and AlkB homolog 5 (ALKBH5) proteins in a mouse model of post-traumatic stress disorder (PTSD). MethodsA PTSD mouse model was established using a single prolonged stress and foot shock stimulation (SPSS) method. The despair, anxiety, and learning and memory functions of PTSD mice were assessed through the open field test, Y-maze test, and forced swimming test. Neuronal damage was detected via HE and Nissl staining. The expression levels of METTL3, FTO, ALKBH5, and neuronal nuclear protein (NEUN) were assessed by Western blot and immunofluorescence staining. ResultsCompared to control group, PTSD mice subjected to SPSS exhibited signs of despair, anxiety, and impaired learning and memory. HE and Nissl staining results showed neuronal damage in the prefrontal cortex of PTSD mice. Western blot and immunofluorescence staining results showed that the expression of the m6A-related proteins METTL3 and FTO decreased, while the expression of ALKBH5 increased in the prefrontal cortex. Additionally, NEUN protein levels showed a declining trend. ConclusionThe pathogenesis of PTSD may be associated with neuronal damage in the prefrontal cortex and alterations in m6A methylation proteins.
4.Effects of different forms of shaker exercises on hyoid muscles activation in healthy adults
Zhenzhen LI ; Qiaoxia ZHEN ; Keke CHEN ; Boyan FANG
Chinese Journal of Tissue Engineering Research 2025;29(18):3791-3797
BACKGROUND:Shaker exercise can enhance the contraction force of the hyoid muscles,which is effective in the treatment of oropharyngeal dysphagia.Shaker exercise should be performed through the main force of the anterior cervical region to train the hyoid muscles.However,patients often experience physical fatigue,leading to compensatory abdominal force exertion.Moreover,the effect of different forms of shaker exercises on the activation of the hyoid muscles in healthy adults is currently unknown.OBJECTIVE:To investigate the effects of shaker exercises with varying force on the activation of the hyoid muscles in healthy adults.METHODS:Twenty healthy adults,including 10 males and 10 females with a mean age of(26.8±3.4)years were recruited as participants for the study.They performed isometric movements of the shaker exercise in the anterior neck and abdominal force modes in sequence.Simultaneously,real-time surface electromyography data were collected from the suprahyoid and infrahyoid muscles and rectus abdominis muscle.At the end of the experiment,participants were asked to rate the physical exertion levels for each type of force using the Borg Rating of Perceived Exertion Scale at the end of the training.Following a series of processing of the original electromyography signals,the peak amplitude and/or average amplitude values of the suprahyoid and infrahyoid muscles and rectus abdominis in the 60-second movement window were calculated and compared,and the differences in the subjective fatigue scores corresponding to the two force modes were compared.RESULTS AND CONCLUSION:The average amplitude of the rectus abdominis muscle in the abdominal force mode was found to be higher than that in the neck force mode(P<0.05).However,there were no significant differences in the mean and peak amplitudes between the suprahyoid and infrahyoid muscles in the two force modes(P>0.05).There was also no significant difference in the Borg Rating of Perceived Exertion Scale scores in the two force modes(P>0.05).These findings suggest that shaker exercises with different force modes(anterior neck and abdomen)have minimal impact on the hyoid muscles and similar feeling of fatigue in healthy adults.
5.The effects of interactive scalp acupuncture and constraint-induced movement therapy for children with spastic hemiplegic cerebral palsy
Xuesong REN ; Zhijun CAI ; Houqin ZHOU ; Keke LI ; Yueqin LI ; Xijun LIU ; Dengna ZHU
Chinese Journal of Physical Medicine and Rehabilitation 2025;47(1):41-45
Objective:To observe any effect of combining interactive scalp acupuncture with constraint-induced movement therapy (CIMT) in treating the upper extremity motor function of children with spastic hemiplegic cerebral palsy.Methods:Fifty-seven children with spastic hemiplegic cerebral palsy were randomly divided into a control group of 28 and an observation group of 29. The control group was given conventional rehabilitation and routine CIMT, while the observation group was additionally provided with interactive scalp acupuncture. In the observation group the CIMT lasted for 30 minutes while the needles were retained. Then, after the needles were removed, the family administered simple rehabilitation training for 2.5 hours guided by a rehabilitation therapist. The treatments were administered 5 days per week for 3 weeks as a session, and the whole treatment consisted of 3 sessions. Before, as well as 1 month and 3 months after the treatment, upper extremity motor function in both groups was evaluated using the Melbourne assessment scale 2 (MA2) and the fine motor Peabody developmental motor scales. Muscle tone in the children′s elbow flexors was assessed using the modified Ashworth scale (MAS).Results:After both 1 and 3 months of treatment, the average MA2 scores of both groups were significantly better than those before treatment, with those of the observation group significantly superior to the control group′s averages. After both 1 and 3 months, both groups′ visual-motor integration scores, grasping ability scores and total scores had improved significantly, with those of the observation group significantly better than in the control group, on average. The MAS scores of both groups were also significantly better than before the treatment, but with no significant differences between the two groups at either time point.Conclusions:Interactive scalp acupuncture combined with constraint-induced movement therapy can significantly improve the upper extremity motor functioning and fine motor ability of children with spastic hemiplegic cerebral palsy.
6.Analysis and prediction of disease burden of idiopathic epilepsy in China
Xiaojun WANG ; Chenwei LI ; Jianglin RAN ; Zhiheng FENG ; Keke YANG ; Huiyuan PENG
Chinese Journal of Neuromedicine 2025;24(7):689-698
Objective:To describe the temporal trend of disease burden of idiopathic epilepsy in China from 1990 to 2021 and predict the incidence of idiopathic epilepsy in China from 2022 to 2035 to provide references for the formulation of relevant health policies and measures.Methods:Based on data from the Global Burden of Disease Study 2021 (GBD 2021) database regarding idiopathic epilepsy in China, changes in disease burden from 1990 to 2021 were acquired. Disease burden was quantified using age-standardized incidence rate (ASIR), age-standardized prevalence rate (ASPR), age-standardized mortality rate (ASMR), age-standardized disability-adjusted life years (DALYs) rate (ASDR) and their 95% uncertain interval (UI). Temporal trend analysis was performed using a linear regression model to estimate the estimated annual percent change (EAPC) and annual percentage change (APC) in incidence of idiopathic epilepsy and their 95% CI. Additionally, incidence and number of patients with idiopathic epilepsy in China from 2022 to 2035 were predicted using Bayesian age-period-cohort model. Results:The ASIR of idiopathic epilepsy increased from 22.35 per 100,000 population in 1990 (95% UI: 15.04-30.92 per 100,000 population) to 28.19 per 100,000 population in 2021 (95% UI: 19.03-37.89 per 100,000 population), with an EAPC of 0.12% (95% CI: -0.10%-0.34%); ASPR of idiopathic epilepsy increased from 189.27 per 100,000 population in 1990 (95% UI: 132.48-252.95 per 100,000 population) to 214.71 per 100,000 population in 2021 (95% UI: 150.10-278.56 per 100,000 population), with an EAPC of -0.32% (95% CI: -0.57%-0.06%); ASMR of idiopathic epilepsy decreased from 1.86 per 100,000 population in 1990 (95% UI: 1.59-2.24 per 100,000 population) to 0.80 per 100,000 population in 2021 (95% UI: 0.67-1.00 per 100,000 population), with an EAPC of -2.96% (95% CI: -3.09%-2.82%); ASDR of idiopathic epilepsy decreased from 178.60 per 100,000 population in 1990 (95% UI: 143.44-220.63 per 100,000 population) to 101.39 per 100,000 population in 2021 (95% UI: 72.51-139.40 per 100,000 population), with an EAPC of -2.38% (95% CI: -2.54%-2.22%). The prediction model showed that by 2035, the prevalence of idiopathic epilepsy in China will be 28.27 per 100,000 (95% CI: 23.19-38.66), with an estimated 394,928 incident cases (95% CI: 324,037-540,128). Conclusions:From 1990 to 2021, the ASIR and ASPR of idiopathic epilepsy in China show an upward trend, while the ASMR and ASDR hace a decline trend. Incidence of idiopathic epilepsy in China is expected to remain stable over the next decade.
7.Caffeic acid phenethyl ester inhibits mGluR5-Fyn signaling to alleviate neuroinflammation and pathological changes in Alzheimer disease
Yuxiang LIU ; Yani HE ; Xueying LIN ; Sihan PENG ; Shuyi LI ; Keke ZHANG ; Wei WEI
Chinese Journal of Pathophysiology 2025;41(5):833-842
AIM:This study aims to investigate the regulatory effects of caffeic acid phenethyl ester(CAPE)on metabotropic glutamate receptor 5(mGluR5)and tyrosine kinase Fyn,and to explore its role in alleviating neuroinflam-mation and pathological features of Alzheimer disease(AD).METHODS:In vitro,the murine neuroblastoma N2a cell line was treated with amyloid β-protein 42 oligomers(Aβ42Os;10 nmol/L to 10 μmol/L)for 24 h.Cell viability was as-sessed by MTT assay.Western blot analyzed mGluR5 expression and Fyn phosphorylation(Tyr416).Pharmacological modulators(CHPG/MPEP)were used to evaluate mGluR5-mediated inflammatory cytokine regulation(qPCR)and Fyn ac-tivation.In vivo,wild-type(WT)and 5×FAD mice(WT,WT+CAPE,5×FAD and 5×FAD+CAPE)were analyzed for AD-related proteins,neuroinflammation(ELISA),glial activation(GFAP/Iba-1 immunofluorescence),and β-amyloid deposi-tion(thioflavin S).RESULTS:(1)Treatment with 1 μmol/L Aβ42Os increased mGluR5 expression(P<0.01)and Fyn phosphorylation(P<0.01)without affecting N2a cell viability.Intracerebral Aβ42Os injection similarly up-regulated hip-pocampal mGluR5 and Fyn(P<0.01).(2)MPEP reduced mGluR5 expression(P<0.01)and Fyn phosphorylation(P<0.01),while suppressing tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)mRNA levels(P<0.01).(3)CAPE decreased mGluR5-Fyn activation in N2a cells,neurons,and 5×FAD mice(P<0.01).(4)CAPE-treated 5×FAD mice exhibited reduced neuroinflammation markers(GFAP,Iba-1,TNF-α,IL-1β,and IL-6),Aβ plaques,and p-APP levels(P<0.01).CONCLUSION:Treatment with CAPE inhibits Aβ42Os-induced mGluR5-Fyn signaling activation,thereby attenuating neuroinflammation and the pathology associated with AD.
8.Mendelian Randomized Study of Protective Effect of Statins on Breast Cancer
Di HU ; Yifang SHUI ; Keke MIAO ; Mengquan LI
Cancer Research on Prevention and Treatment 2025;52(2):165-171
Objective To genetically investigate the protective effects of statins on breast cancer. Methods Instrumental variables for the statin target gene HMGCR and five other cholesterol-regulated genes (LDLR, PCSK9, ABCG8, APOB, and NPC1L1) were obtained from previous expression quantitative trait locus (eQTL) studies. Cholesterol-regulated genes predicted by these instrumental variables served as the exposure factors. Mendelian randomization based on pooled data (SMR) was conducted to explore the genetic effects of exposure factors on the incidence risk of all breast cancers, ER+ breast cancer, and ER-breast cancer. Instrumental variables for total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and non-high-density lipoprotein cholesterol (non-HDL-C) were derived from a previous human genome-wide association study and restricted to be chromosomally located within 100 kb of the above cholesterol regulatory genes; the instrumental variables could predict TC, LDL-C, or non-HDL-C levels under the regulation of the abovementioned cholesterol-associated genes which were used as exposure factors. Two-sample Mendelian randomization (IVW, MR-PRESSO, and MR-Egger) was used to explore the genetic effects of exposure factors on the risk of all breast cancers, ER+ breast cancer, and ER− breast cancer. Results SMR analysis reported that elevated HMGCR expression was significantly associated with the increased incidence risk of all breast cancers and ER+ breast cancer (P=0.044 and P=0.039, respectively) but not with the change in incidence risk of ER− breast cancer (P=0.190); the other five regulatory genes were not significantly correlated with the change in incidence risk of all breast cancers, ER+ breast cancer, and ER− breast cancer (all P>0.05). IVW analysis reported that under the regulation of HMGCR, elevated levels of peripheral TC, LDL-C, and non-HDL-C significantly increased the incidence risk of all breast cancers (P=1.160e-05, P=1.248e-05, and P=1.869e-05) and the incidence risk of ER+ breast cancer (P=3.181e-04, P=2.231e-04, and P=3.520e-04), but they were not associated with a change in the incidence risk of ER− breast cancer (P=0.062, P=0.133, and P=0.055). The results of MR-PRESSO and MR-Egger analyses supported the IVW results. Conclusion Statins could reduce the incidence risk of ER+ breast cancer at the genetic level, but there is no such protective effects on ER− breast cancer.
9.Inhibitory effect of safranal on proliferation,migration and phenotypic transformation of vascular smooth muscle cells of rats induced by high glucose in vitro
Yixuan GAO ; Peng WANG ; Silong ZHANG ; Ruijuan GAO ; Yingfang MA ; Keke ZHANG ; Dan FENG ; Zongqi HUANG ; Ketao MA ; Li LI ; Junqiang SI
Journal of Jilin University(Medicine Edition) 2025;51(4):948-957
Objective:To discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.
10.Textual Research and Clinical Application Analysis of Classic Formula Fangji Fulingtang
Xiaoyang TIAN ; Lyuyuan LIANG ; Mengting ZHAO ; Jialei CAO ; Lan LIU ; Keke LIU ; Bingqi WEI ; Yihan LI ; Jing TANG ; Yujie CHANG ; Jingwen LI ; Bingxiang MA ; Weili DANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):270-277
The classic formula Fangji Fulingtang is from ZHANG Zhongjing's Synopsis of the Golden Chamber in the Eastern Han dynasty. It is composed of Stephaniae Tetrandrae Radix, Astragali Radix, Cinnamomi Ramulus, Poria, and Glycyrrhizae Radix et Rhizoma, with the effects of reinforcing Qi and invigorating spleen, warming Yang and promoting urination. By a review of ancient medical books, this paper summarizes the composition, original plants, processing, dosage, decocting methods, indications and other key information of Fangji Fulingtang, aiming to provide a literature basis for the research, development, and clinical application of preparations based on this formula. Synonyms of Fangji Fulingtang exist in ancient medical books, while the formula composition in the Synopsis of the Golden Chamber is more widespread and far-reaching. In this formula, Stephaniae Tetrandrae Radix, Astragali Radix, Cinnamomi Ramulus, Poria, and Glycyrrhizae Radix et Rhizoma are the dried root of Stephania tetrandra, the dried root of Astragalus embranaceus var. mongholicus, the dried shoot of Cinnamomum cassia, the dried sclerotium of Poria cocos, and the dried root and rhizome of Glycyrrhiza uralensis, respectively. Fangji Fulingtang is mainly produced into powder, with the dosage and decocting method used in the past dynasties basically following the original formula. Each bag is composed of Stephaniae Tetrandrae Radix 13.80 g, Astragali Radix 13.80 g, Cinnamomi Ramulus 13.80 g, Poria 27.60 g, and Glycyrrhizae Radix et Rhizoma 9.20 g. The raw materials are purified, decocted in water from 1 200 mL to 400 mL, and the decoction should be taken warm, 3 times a day. Fangji Fulingtang was originally designed for treating skin edema, and then it was used to treat impediment in the Qing dynasty. In modern times, it is mostly used to treat musculoskeletal and connective tissue diseases and circulatory system diseases, demonstrating definite effects on various types of edema and heart failure. This paper clarifies the inheritance of Fangji Fulingtang and reveals its key information (attached to the end of this paper), aiming to provide a theoretical basis for the development of preparations based on this formula.

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