1.Construction and Application Evaluation of an Integrated Traditional Chinese and Western Medicine Risk Prediction Model for Readmission in Patients with Stable Angina of Coronary Heart Disease:A Prospective Study Based on Real-World Clinical Data
Wenjie HAN ; Mingjun ZHU ; Xinlu WANG ; Rui YU ; Guangcao PENG ; Qifei ZHAO ; Jianru WANG ; Shanshan NIE ; Yongxia WANG ; Jingjing WEI
Journal of Traditional Chinese Medicine 2025;66(6):604-611
ObjectiveBy exploring the influencing factors of readmission in patients with stable angina of coronary heart disease (CHD) based on real-world clinical data, to establish a risk prediction model of integrated traditional Chinese and western medicine, in order to provide a basis for early identification of high-risk populations and reducing readmission rates. MethodsA prospective clinical study was conducted involving patients with stable angina pectoris of CHD, who were divided into a training set and a validation set at a 7∶3 ratio. General information, traditional Chinese medicine (TCM)-related data, and laboratory test results were uniformly collected. After a one-year follow-up, patients were classified into a readmission group and a non-readmission group based on whether they were readmitted. Univariate and multivariate logistic regression analyses were performed to identify independent risk factors for readmission. A risk prediction model of integrated traditional Chinese and western medicine was constructed and visualized using a nomogram. The model was validated and evaluated in terms of discrimination, calibration, and clinical decision curve analysis. ResultsA total of 682 patients were included, with 477 in the training set and 205 in the validation set, among whom 89 patients were readmitted. Multivariate logistic regression analysis identified heart failure history [OR = 6.93, 95% CI (1.58, 30.45)], wiry pulse [OR = 2.58, 95% CI (1.42, 4.72)], weak pulse [OR = 3.97, 95% CI (2.06, 7.67)], teeth-marked tongue [OR = 4.38, 95% CI (2.32, 8.27)], blood stasis constitution [OR = 2.17, 95% CI (1.06, 4.44)], phlegm-stasis mutual syndrome [OR = 3.64, 95% CI (1.87, 7.09)], and elevated non-high-density lipoprotein cholesterol [OR = 1.30, 95% CI (1.01, 1.69)] as influencing factors of readmission. These factors were used as predictors to construct a nomogram-based risk prediction model for readmission in patients with stable angina. The model demonstrated moderate predictive capability, with an area under the receiver operating characteristic curve (AUC) of 0.818 [95% CI (0.781, 0.852)] in the training set and 0.816 [95% CI (0.779, 0.850)] in the validation set. The Hosmer-Lemeshow test showed good calibration (χ² = 4.55, P = 0.80), and the model's predictive ability was stable. When the threshold probability exceeded 5%, the clinical net benefit of using the model to predict readmission risk was significantly higher than intervening in all patients. ConclusionHistory of heart failure, teeth-marked tongue, weak pulse, wiry pulse, phlegm-stasis mutual syndrome, blood stasis constitution, and non-high-density lipoprotein cholesterol are influencing factors for readmission in patients with stable angina of CHD. A clinical prediction model was developed based on these factors, which showed good discrimination, calibration, and clinical utility, providing a scientific basis for predicting readmission events in patients with stable angina.
2.Association between Mediterranean diet scores and dental caries among children and adolescents with neurodevelopmental disorders
XIONG Wenjuan, SU Yuanyuan, LIU Zhao, HUANG Xiaoqing, QU Zhiyi, CUI Shanshan
Chinese Journal of School Health 2025;46(2):172-176
Objective:
To explore the association between mediterranean diet (MD) patterns and dental caries among children and adolescents with neurodevelopmental disorders (NDD), so as to provide a basis for developing scientific anti caries strategies related to diet.
Methods:
From December 2021 to June 2024, a questionnaire survey, a three day 24 hour dietary review survey, oral health examination, physical development measurement and Childhood Autism Rating Scale (CARS) evaluation were conducted involving 147 children and adolescents aged 2-22 years with NDD from nine special education schools and rehabilitation institutions in Tianjin. Group comparisons were carried out using the Mann-Whitney U test, Chi-square test, or Fisher s exact probability method. The correlation between dietary quality and dental caries was analyzed by adopting multiple linear regression analysis and restricted cubic spline.
Results:
There were 46 children and adolescents (31.3%) in the non dental caries group and 101 children and adolescents (68.7%) in the dental caries group. The number of decayed missing and filled teeth (dmft) was 2.0 (4.0), and the MD score was 4.0 (2.0) points. There were 62 children and adolescents (42.2%) in the low MD scores group and 85 children and adolescents (57.8%) in the high MD scores group. There was no significant difference in MD scores between NDD children in the non dental caries group and those in the dental caries group [nondental caries group:4.0(2.0), dental caries group:4.0(2.0), Z= -0.14, P >0.05]. The MD scores and dmft exhibited increasing and then decreasing trend ( P total =0.02, P non lineary = 0.04 ). Children and adolescents with NDD in the MD high scores group had a lower number of dmft than those in the MD low scores group ( β= -2.00 , 95%CI =-3.39 to -0.62, P <0.05). However, in children and adolescents with NDD and CARS scores ≥30, the above association was insignificant ( β=-0.63, 95%CI=-0.29-0.15, P >0.05).
Conclusion
Children and adolescents with NDD who have dietary patterns similar to the Mediterranean diet, are found to have fewer dental caries, and this is observed among those with no or mild symptoms of autism spectrum disorder.
3.Accuracy of large language models for answering pediatric preventive dentistry questions
GUAN Boyan ; XU Minghe ; ZHANG Huiqi ; MA Shulei ; ZHANG Shanshan ; ZHAO Junfeng
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(4):313-319
Objective:
To evaluate and compare the accuracy of responses to pediatric preventive dentistry-related questions between the domestic large language model, ChatGLM-6B, and the international large language model, ChatGPT-3.5, in order to provide insights for further research and development of domestic language models in the field of oral medicine.
Methods:
A total of 100 common pediatric preventive dentistry questions of varying difficulty levels [basic (n = 35), intermediate (n = 35), and advanced (n = 30) ] were provided by pediatric preventive dentistry experts. Two doctors independently registered these questions with ChatGPT-3.5 and ChatGLM-6B and collected the answers. A cohort of 16 dentists assessed responses generated by ChatGLM-6B and ChatGPT-3.5 using a predefined 3-point Likert scale. The average score of the ratings from 16 doctors was taken as the answer score. If the answer score was higher than 2.8, it was accepted as a accurate answer; if the score was lower than 1.4, it was accepted as an inaccurate answer; if the score was between 1.4 and 2.8, it was accepted as a partially accurate answer. Comparative analysis was conducted on the accuracy rates and evaluation outcomes between the two groups. Consistency analysis of the ratings was conducted
Results:
The answer accuracy rates of ChatGPT-3.5 and ChatGLM-6B for 100 pediatric preventive dentistry questions were comparable: ChatGPT-3.5 demonstrated 68% accurate, 30% partially accurate, and 2% inaccurate responses, while ChatGLM-6B showed 67% accurate, 31% partially accurate, and 2% inaccurate responses, with no statistically significant differences (P>0.05). Both models exhibited equivalent accuracy across questions of varying difficulty levels (basic, intermediate, advanced), showing no statistical differences (P>0.05). The overall average scores for ChatGPT3.5 and ChatGLM-6B in answering all questions were both 2.65, with no statistically significant difference (P>0.05). For questions of different difficulty levels, ChatGPT3.5 had an average score of 2.66 for basic questions while ChatGLM-6B had an average score of 2.70. For intermediate questions, ChatGPT3.5 had an average score of 2.63 and ChatGLM-6B had an average score of 2.64. For advanced questions, ChatGPT3.5 had an average score of 2.68, and ChatGLM-6B had an average score of 2.61. No statistically significant differences were observed across any difficulty category (P>0.05). The consistency of the experts’ grading ranged from fair to moderate.
Conclusion
This study demonstrates the potential of both ChatGLM-6B and ChatGPT-3.5 in answering pediatric preventive dentistry questions. ChatGLM-6B performed similarly to ChatGPT-3.5 in this field, but the accuracy rates of both models fell short of expectations and are not suitable for clinical use. Future efforts should focus on improving the accuracy and consistency of large language models in providing medical information, as well as developing specialized medical models for the field of oral medicine.
4.Association between ambient ozone exposure during pregnancy and risk of preterm birth in Guangdong Province
Peng HU ; Shanshan RAN ; Qingmei LIN ; Yin YANG ; Zilong ZHANG ; Xiaoling GUO ; Yonggui GAO ; Jinde ZHAO ; Hualiang LIN
Journal of Environmental and Occupational Medicine 2025;42(4):379-384
Background Air pollution exposure has a significant impact on maternal and child health. However, the research on the association between ambient ozone (O3) exposure during pregnancy and the risk of premature birth in newborns is limited, and the conclusions are inconsistent. Objective To investigate the association of ambient O3 exposure during pregnancy with the risk of preterm birth in Guangdong Province. Methods Data of pregnant women in Guangzhou from 2013 to 2019 and Foshan from 2018 to 2023 were collected, and O3 concentrations during different trimesters were assessed according to maternal residential addresses. Bilinear interpolation was used to evaluate the concentrations of air pollution. A cohort study design was adopted in our study. Restricted cubic spline curves were used to evaluate the exposure-response relationship between O3 exposure and preterm birth risk and explore potential exposure threshold of O3. Logistic regression models were used to evaluate the association of O3 exposure with preterm birth. Results A total of 702 924 pregnant women were included in this study, of whom 43 051 (6.12%) were preterm. The average O3 exposure concentrations of pregnant women during the first, second, third, and whole trimesters were 95.51, 97.51, 100.60, and 97.87 μg·m−3, respectively. We observed J-shaped associations between O3 exposure and preterm birth risk during the second, third, and whole trimesters of pregnancy using restricted cubic spline curves. This study found that there were threshold concentrations between O3 exposure and preterm birth risk during different gestational periods, and the threshold concentrations in the first, second, third, and whole trimesters were 112.32, 99.83, 111.74, and 112.46 μg·m−3, respectively. During the second, third, and whole trimesters of pregnancy, after adjusting for maternal age, baby sex, pre-pregnancy body mass index, mode of delivery, baby birth weight, gestational diabetes, and gestational hypertension, the odds ratios (OR) of preterm birth were 1.02 (95%CI: 1.01, 1.04), 1.02 (95%CI: 1.00, 1.03), and 1.17 (95%CI: 1.13, 1.21) for each 10 μg·m−3 increase in O3 concentration above the O3 threshold. No significant association was found between O3 exposure and the risk of preterm birth during the first trimester. Conclusion There is a nonlinear association between the risk of preterm birth and O3 exposure during pregnancy, and higher concentrations of O3 exposure during pregnancy are associated with the risk of preterm birth. Above the O3 threshold concentration during pregnancy, especially during the second, third, and whole trimesters, the risk of preterm birth elevates with the increase of O3 exposure concentrations.
5.Quality Evaluation of Black Panacis Quinquefolii Radix Based on Neuroprotective Spectrum-effect Relationship
Yuting YANG ; Shuyun LIANG ; Shanshan LI ; Yulong YANG ; Ziqi YANG ; Guangzhi CAI ; Liru ZHAO ; Jiyu GONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):149-156
ObjectiveTo clarify the neuroprotective effect of black Panacis Quinquefolii Radix(PQR) and explore its active ingredients, with the aim of establishing an activity-oriented quality evaluation method. MethodsTransgenic Tg(HuC∶EGFP) zebrafish was used to establish a neuronal injury model by aluminum chloride immersion. Different doses(10, 20 mg·L-1) of PQR and black PQR ethanol extracts were administered. The neuroprotective effects of PQR and black PQR were compared by analyzing the fluorescent area and intensity of zebrafish neurons. Based on ultra-performance liquid chromatography(UPLC), a fingerprint profile of black PQR was established, followed by principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Differential components were screened using the criteria of variable importance in the projection(VIP) value>1 and P<0.05. The neuroprotective activity of 14 batches of black PQR was assessed, and Spearman correlation analysis was used to identify saponins related to neuroprotective activity, which were then validated. Based on the above results, active marker components were determined, and an UPLC method was established for their quantitation with clear content limits. ResultsPharmacological efficacy results showed that both PQR and black PQR at different doses could significantly improved neuronal damage in zebrafish. At a dose of 20 mg·L-1, black PQR demonstrated superior efficacy(P<0.05). The fingerprint similarities of 14 batches of black PQR were>0.94, with 26 common peaks identified. Through comparison with the reference standards, 8 components were confirmed, including peak 1(ginsenoside Rg1), peak 2(ginsenoside Re), peak 5(ginsenoside Rb1), peak 9(ginsenoside Rd), peak 16[ginsenoside 20(S)-Rg3], peak 17[ginsenoside 20(R)-Rg3], peak 18(ginsenoside Rk1), and peak 19(ginsenoside Rg5). The results of PCA and OPLS-DA indicated that there were differences in saponins among black PQR samples from different origins, and 12 differential components were screened. All 14 batches of black PQR exhibited good protective effects on zebrafish neurons, with Shaanxi-produced black PQR showing superior protective effects compared to the other three production regions. Spearman correlation analysis revealed that a total of 11 components, including ginsenosides 20(S)-Rg3, 20(R)-Rg3, Rk1 and Rg5, showed a significant positive correlation with the neuroprotective effect in zebrafish(P<0.05). The activity validation results indicated that ginsenosides 20(S)-Rg3, 20(R)-Rg3, Rk1 and Rg5 were the primary components responsible for the neuroprotective effects of black PQR. Quantitative analysis showed that the content of ginsenoside 20(S)-Rg3 in 14 batches of black PQR ranged from 0.17% to 0.52%, and the repair rate of neuronal damage ranged from 42.77% to 97.83%. ConclusionBased on the fingerprint and neuronal protective activity, the spectrum-effect related quality control model of black PQR was established, with ginsenoside 20(S)-Rg3 as the quality control index, and the neuronal damage repair rate≥60% as the evaluation standard, the minimum limit of ginsenoside 20(S)-Rg3 in black PQR should be≥0.20%.
6.Clinical and Mechanistic Study of Modified Sinisan in Treating Precancerous Lesions of Digestive System Based on "Inflammation-to-Cancer Transformation"
Xuhang SUN ; Dandan WEI ; Xin PENG ; Shanshan LI ; Yihan ZHAO ; Fuke YAO ; Shiqing JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):221-234
Tumorigenesis, invasion, and metastasis occur in the context of a persistent inflammatory microenvironment, and a variety of inflammatory factors can lead to the development of various tumors. Guided by the thought of "preventive treatment of disease" in TCM and the concept of tertiary prevention in modern medicine, it is of great significance to effectively intervene in the inflammatory stage of the disease, interrupt disease progression, prevent the occurrence of malignant tumors, and reverse the process of "inflammation-to-cancer transformation". Sinisan, a commonly used prescription in the Treatise on Febrile Diseases, has been widely applied in the treatment of precancerous lesions of the digestive system, demonstrating considerable advantages. This article reviewed literature from the past 20 years, summarizing the application of Sinisan in precancerous lesions of the digestive system from three aspects: the exploration of its prescription-syndrome relationship, clinical application, and mechanistic study. It is found that basic syndrome indications of Sinisan include harmonizing the Earth element to promote spleen-stomach transportation and transformation, soothing the liver and nourishing the Wood element to restore the smooth flow of Qi, and regulating Yin and Yang to relieve stagnation within the system. In clinical application, Sinisan has shown significant efficacy in atrophic gastritis and precancerous conditions such as intestinal metaplasia, gastric ulcer, ulcerative colitis, esophagitis, and pancreatitis. Mechanistic studies have revealed that Sinisan can inhibit inflammatory factors and improve the inflammatory microenvironment, inhibit cell proliferation and regulate apoptosis, exhibit anti-angiogenic and antitumorigenic effects, modulate immune function, and exert antioxidant effects. These mechanisms can be achieved by regulating pathways such as nuclear factor erythroid 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1), farnesoid X receptor (FXR)/Nrf2, phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), Takeda G protein-coupled receptor 5/cyclic adenosine monophosphate/protein kinase A (TGR5/cAMP/PKA), interleukin-4/signal transducer and activator of transcription 6 (IL-4/STAT6), Janus kinase/signal transducer and activator of transcription (JAK/STAT), RhoA/Rho-associated protein kinase (RhoA/ROCK), and transforming growth factor-β/Smad proteins (TGF-β/Smads), confirming Sinisan's role in reversing the inflammation-to-cancer transformation. The current research status of Sinisan in precancerous lesions of the digestive system was thoroughly examined through the above three aspects, along with the identification of limitations and areas for improvement in current research. The aim is to provide a basis and support for future in-depth research on Sinisan, promote the development of new integrated treatment models combining TCM and Western medicine for precancerous lesions, and aid in the research and development of drugs related to precancerous lesions.
7.Consistency Evaluation of Processing of Black Panacis Quinquefolii Radix Decoction Pieces Based on "Chromaticity-chemistry-activity"
Yulong YANG ; Shanshan LI ; Yuting YANG ; Ziqi YANG ; Guangzhi CAI ; Liru ZHAO ; Jiyu GONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):195-203
ObjectiveTo assess the quality consistency of black Panacis Quinquefolii Radix(bPQR) decoction pieces prepared by atmospheric and pressurized steaming processes based on chromaticity-chemical composition-vasoactive inhibition. The ultimate goal was to screen the pressurized steaming process yielding quality equivalent to atmospheric steaming, and optimize the processing technology of bPQR. MethodsThe bPQR decoction pieces were prepared using both atmospheric and pressurized steaming processes, and the chromaticity values[lightness value(L*), red/green chromaticity value(a*), yellow/blue chromaticity value(b*), total chromaticity value(E*ab)] were measured. High performance liquid chromatography(HPLC) was employed to establish fingerprint profiles for the decoction pieces, and cluster analysis was conducted on chromaticity values and the common peak areas in fingerprint profiles to elucidate the quality relationships between the decoction pieces processed by different methods. The optimal atmospheric steaming of bPQR decoction pieces was determined through zebrafish angiogenesis inhibition experiments. The contents of ginsenosides Rg1, Re, Rb1, 20(S)-Rg3, Rk1 and Rg5 in the decoction pieces were quantified, and Spearman correlation analysis was employed to investigate the relationship between saponin content, chromaticity, and angiogenesis inhibition activity during the steaming process. By integrating the consistency of chromaticity, saponin components and angiogenesis inhibition activity, pressurized steaming conditions with quality equivalent to the atmospheric pressure method were selected. ResultsCompared with the atmospheric steaming method, pressurized steaming resulted in faster color darkening and higher conversion rates of ginsenosides in bPQR decoction pieces. Moreover, the neovascularization inhibitory activity of bPQR decoction pieces continued to increase with the deepening of processing. Based on the effectiveness and safety, the optimal process for preparing bPQR decoction pieces with neovascularization inhibitory activity was determined to be atmospheric steaming for 21 h. All six ginsenosides tested exhibited strong to extremely strong correlations with both the chromaticity values of the decoction pieces and their neovascularization inhibitory activities. Among them, ginsenosides Rg1, Re and Rb1 exhibited positive correlations with chromaticity values and negative correlations with zebrafish angiogenesis inhibition activity. Conversely, ginsenosides 20(S)-Rg3, Rk1 and Rg5 showed negative correlations with chromaticity values and positive correlations with zebrafish angiogenesis inhibition activity. By integrating chromaticity values, cluster analysis results, as well as the results of activity, it was determined that the quality of bPQR decoction pieces steamed under pressurized conditions of 110 ℃(0.045 MPa) for 5 h and 115 ℃(0.07 MPa) for 3 h was highly consistent with that obtained by atmospheric steaming for 21 h. ConclusionThe preparation of bPQR decoction pieces by pressurized steaming has the advantages of short preparation time, low energy consumption, and rapid saponin conversion rate, making it a viable alternative to atmospheric steaming for preparing bPQR decoction pieces. Meanwhile, the evaluation method based on chromaticity-chemical composition-activity can provide a more scientific and effective explanation of change rules in the quality during traditional Chinese medicine processing, and offer a new model for optimizing processing technology and enhancing quality control.
8.Compatibility Effect and Mechanism of Astragali Radix-Curcumae Rhizoma on Antitumor: A Review
Hanyu KANG ; Shanshan LI ; Dandan WEI ; Yihan ZHAO ; Ruxin DU ; Shiqing JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):276-286
Malignant tumor is a serious and difficult disease threatening human health, which has a high morbidity and mortality rate worldwide. Traditional Chinese medicine has unique advantages in improving the therapeutic effect of malignant tumors and alleviating adverse reactions. Traditional Chinese medicine believes that Qi deficiency and blood stasis are important pathogeneses in the development of malignant tumors, and the method of supplementing Qi and activating blood is an effective strategy for treating malignant tumors. Astragali Radix, sweet in taste and warm in nature, has effects of tonifying Qi and rising Yang, strengthening the exterior and reducing sweat, promoting fluid and nourishing blood. Curcumae Rhizoma, acrid and bitter in taste and warm in nature, has the effects of promoting Qi and breaking blood stasis, eliminating mass, and relieving pain. Astragali Radix-Curcumae Rhizoma, as the classic herb pair of invigorating Qi and activating blood, has a clear effect on inhibiting tumor growth and metastasis. Studies have shown that Astragali Radix-Curcumae Rhizoma contains astragalus polysaccharide, astragaloside, calycosin, formononetin, curcumin, β-elemene, curcumenol, curcumenone, curcumendione, gemacrone, and other anti-tumor active ingredients. It can significantly inhibit the occurrence and development of liver cancer, colorectal cancer, gastric cancer, lung cancer, ovarian cancer, cervical cancer, breast cancer, and other cancers and has the advantages of superposition effect, synergistic complementarity, and increased dissolution compared with single herb and monomer of Chinese traditional herbs and has been widely valued in the field of TCM anti-cancer. Its anti-tumor mechanism includes inhibition of tumor cell proliferation, promotion of tumor cell apoptosis and autophagy, anti-invasion and metastasis, regulation of immune function, and enhancement of anti-tumor drug sensitivity. By combining Chinese and foreign literature, the compatibility effect and anti-tumor mechanism of Astragali Radix-Curcumae Rhizoma were summarized, and then scientific compatibility of these two herbs was expounded, in order to provide a useful reference for clinical application and future research of Astragali Radix-Curcumae Rhizoma.
9.Therapeutic Effect of Cranial Painkiller Pills' Extract Powder in Treatment of Trigeminal Neuralgia Induced by Injection of Talci Pulvis into Infraorbital Foramen of Model Rats Based on OTULIN-regulated Neuroinflammation
Shuran LI ; Xinwei WANG ; Jing SUN ; Dan XIE ; Ronghua ZHAO ; Lei BAO ; Zihan GENG ; Qiyue SUN ; Jingsheng ZHANG ; Yaxin WANG ; Xihe CUI ; Xinying LI ; Bing HAN ; Tianjiao LU ; Xiaolan CUI ; Liying LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):21-28
ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis, evaluate its potential clinical application value, and compare the therapeutic effect with that of Cranial Painkiller granules, so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis, and the rats were randomly divided into the normal group, model group, carbamazepine group (60 mg·kg-1), Cranial Painkiller granules group (2.70 g·kg-1), and low, medium, and high dosage groups of Cranial Painkiller pills' extract powder (1.35, 2.70, 5.40 g·kg-1) according to the basal mechanical pain thresholds, and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling, and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin (HE) staining was used to detect pathological changes in the trigeminal ganglion, and enzyme-linked immunosorbent assay (ELISA) was used to detect the inflammatory factors interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) levels in rat serum, as well as neuropeptide substance P (SP) and β-endorphin (β-EP) levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3, ASC, Caspase-1, and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group, the pain threshold of rats in the model group showed a continuous significant decrease (P<0.01). The pathological damage of brain tissue was significant (P<0.01), and the inflammatory levels of IL-1, IL-6, IL-8, and TNF-α in serum were significantly elevated (P<0.01). The level of the SP in the brain tissue was significantly elevated (P<0.01), and the level of β-EP was significantly reduced (P<0.01), while the level of OTULIN was significantly reduced, and NLRP3, ASC, and Caspase-1 protein levels were significantly elevated (P<0.01). After administration of the drug, compared with the model group, the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased (P<0.01). The inflammatory levels of IL-1, IL-6, IL-8, and TNF-α and SP levels significantly decreased (P<0.01), and the β-EP levels were significantly elevated (P<0.01), while the levels of OTULIN protein were significantly elevated (P<0.05, P<0.01), and the levels of NLRP3, ASC proteins were decreased (P<0.01)in high dose Cranial Painkiller pills' extract powder. Meanwhile, compared with those in the model group, the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement (P<0.05, P<0.01). ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis, and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.
10.Research on Regulatory Mechanism of Verbenalin on HCoV-229E-infected Macrophage Injury Based on Mitophagy
Qiyue SUN ; Lei BAO ; Zihan GENG ; Ronghua ZHAO ; Shuran LI ; Xihe CUI ; Jingsheng ZHANG ; Xian LIU ; Rui XIE ; Xiaolan CUI ; Shanshan GUO ; Jing SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):29-37
ObjectiveTo investigate the protective effect and mechanism of verbenalin on mouse mononuclear macrophage leukemia cells (RAW264.7) damaged by human coronavirus (HCoV)-229E infection, thereby providing experimental evidence for its development and application. MethodsRAW264.7 macrophages were infected with different concentrations of HCoV-229E to establish a coronavirus-induced macrophage injury model using the cell counting kit-8 (CCK-8) assay for assessing cell proliferation and viability. Cells were randomly divided into four groups: normal control, verbenalin group (125 μmol·L-1), model group (HCoV-229E), and HCoV-229E + verbenalin group (HCoV-229E + 125 μmol·L-1 verbenalin). Cell viability was measured using the CCK-8 assay, and the maximum non-toxic concentration (CC0), half-maximal cytotoxic concentration (CC50), half-maximal effective concentration (EC50), and selectivity index (SI) of verbenalin were calculated. Calcein/PI double staining was used to assess cell viability and cytotoxicity, and JC-1 staining was applied to evaluate changes in mitochondrial membrane potential (MMP). mito-Keima adenovirus labeling was used to assess mitophagy levels in each group. ResultsA macrophage infection model was successfully established by infecting RAW264.7 cells with the original concentration of HCoV-229E for 36 h. The CC0 of verbenalin was 125 μmol·L-1. The CC50 was 448.25 μmol·L-1. The EC50 against HCoV-229E-infected cells was 46.28 μmol·L-1, and the SI was 9.68. Compared with the normal group, the model group showed significantly reduced cell survival rate (P<0.01), increased cell death rate (P<0.01), decreased MMP (P<0.01), and suppressed mitophagy (P<0.01). In contrast, verbenalin treatment significantly improved cell survival rate (P<0.01), reduced cell death rate (P<0.01), alleviated MMP loss (P<0.01), and enhanced mitophagy levels (P<0.01) compared with the model group. ConclusionVerbenalin can enhance the survival rate of macrophages following HCoV-229E infection. The underlying mechanism may be associated with the activation of mitophagy, maintenance of MMP stability, and alleviation of mitochondrial damage.


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