1.Historical Evolution and Key Information Research on Classic Formula Puji Xiaoduyin
Lianchao ZHU ; Lyuyuan LIANG ; Jing TANG ; Jialei CAO ; Ziming XU ; Huizhen ZHANG ; Zhidan GUO ; Rongze MA ; Zhengshao ZHANG ; Bingqi WEI ; Xiubo DU ; Bingxiang MA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):238-247
Puji Xiaoduyin, a specialized formula for the swollen-head epidemic, was recorded in the Catalogue of Ancient Classical Formula (the Second Batch)-Han Medicine, published in September 2023. It had been inherited and developed by medical experts of successive generations and passed down to this day. This paper sorted out the historical evolution of this formula using bibliometric methods. It also comprehensively analyzed key information on the formula name, historical origin, drug dosage, herb origin, processing methods, decocting methods, function, and clinical applications. Additionally, this paper analyzed the application of this formula in both modern and ancient times. Results showed that the formula was first recorded as "Puji Xiaodu Yinzi" in LI Dongyuan's Proven Formulas written by LI Gao from the Jin dynasty. The medicinal composition and dosage were: Scutellariae Radix and Coptidis Rhizoma (20.65 g each), Ginseng Radix et Rhizoma 12.39 g, Scrophulariae Radix, Citri Reticulatae Pericarpium, and Glycyrrhizae Radix et Rhizoma (8.26 g each), Forsythiae Fructus, Arctii Fructus, Isatidis Radix, and Lasiosphaera Calvatia (4.13 g each), Bombyx Batryticatus and Cimicifugae Rhizoma (2.891 g each), Bupleuri Radix and Platycodonis Radix (8.26 g each). These medicines were grounded to fine powder. One dose, including 20.65 g of the powder, was mixed with 600 mL of water and decocted to 300 mL. After abandoning slag, the medicine should be taken warm frequently. In the formula, Bombyx Batryticatus is stir-fired. With the effect of dispersing wind and clearing heat, removing stagnation and dissipating mass, the formula is specialized in swollen-head epidemic, pestilence, red and swelling head, face, and neck, dry mouth and tongue, as well as other diseases resulting from toxic heat stagnated in the upper jiao. The formula is widely used in treating diseases involving the respiratory, dermal, ophthalmologic, otolaryngologic, and nervous systems. The formula is most frequently used for respiratory diseases, with a wide range of symptoms including parotitis/mumps (66 times), followed by tonsillitis (28 times). In conclusion, the broadly applied formula has accurate efficacy and great development value.
2.Analysis of Risk Factors and Establishment of Prediction Model for Turbidity Toxicity Accumulation Syndrome in Patients with Chronic Atrophic Gastritis
Yican WANG ; Chenggong ZHAO ; Pengli DU ; Jie WANG ; Yuxi GUO ; Haiyan BAI ; Yongli HUO ; Xiaomeng LANG ; Zheng ZHI ; Bolin LI ; Jianping LIU ; Yanru CAI ; Jianming JIANG ; Qian YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):288-295
ObjectiveThis paper aims to explore the risk factors for chronic atrophic gastritis (CAG) with turbidity toxin accumulation syndrome and establish a prediction model. MethodsClinical data of 180 patients with CAG who participated in the "clinical study of Xianglian Huazhuo Particles blocking CAG cancer transformation" of Hebei Sheng Zhong Yi Yuan from July 2021 to March 2022 were collected. After confounding factors were controlled by propensity score matching, patients were divided into a training set (namely dev) and a validation set (namely vad) in a seven to three ratio. The risk factors for CAG with turbidity toxin accumulation syndrome in the training set were investigated by using univariate Logistic regression analysis and least absolute shrinkage and selection operator (namely Lasso) regression algorithms. Subsequently, a model, named model 1se, was developed by using the training set data to predict the risk factors for CAG with turbidity toxin accumulation syndrome. The accuracy of the prediction model was assessed by using various methods, including the receiver operating characteristic (ROC) curve, Hosmer-Lemeshow test (H-L), calibration plot, and decision curve analysis (DCA). ResultsAge, body mass index (BMI), family history of cancer, job and life satisfaction, yellow and greasy fur with slippery pulse, and heavy body sensation were independent risk factors of the model. The prediction model showed excellent predictive value for both the training and validation sets. ConclusionThe established prediction model for CAG with turbidity toxin accumulation syndrome has high discrimination and excellent calibration, which could provide an excellent clinical basis for disease diagnosis and individualized treatment of patients.
3.Historical Evolution and Key Information Research on Classic Formula Puji Xiaoduyin
Lianchao ZHU ; Lyuyuan LIANG ; Jing TANG ; Jialei CAO ; Ziming XU ; Huizhen ZHANG ; Zhidan GUO ; Rongze MA ; Zhengshao ZHANG ; Bingqi WEI ; Xiubo DU ; Bingxiang MA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):238-247
Puji Xiaoduyin, a specialized formula for the swollen-head epidemic, was recorded in the Catalogue of Ancient Classical Formula (the Second Batch)-Han Medicine, published in September 2023. It had been inherited and developed by medical experts of successive generations and passed down to this day. This paper sorted out the historical evolution of this formula using bibliometric methods. It also comprehensively analyzed key information on the formula name, historical origin, drug dosage, herb origin, processing methods, decocting methods, function, and clinical applications. Additionally, this paper analyzed the application of this formula in both modern and ancient times. Results showed that the formula was first recorded as "Puji Xiaodu Yinzi" in LI Dongyuan's Proven Formulas written by LI Gao from the Jin dynasty. The medicinal composition and dosage were: Scutellariae Radix and Coptidis Rhizoma (20.65 g each), Ginseng Radix et Rhizoma 12.39 g, Scrophulariae Radix, Citri Reticulatae Pericarpium, and Glycyrrhizae Radix et Rhizoma (8.26 g each), Forsythiae Fructus, Arctii Fructus, Isatidis Radix, and Lasiosphaera Calvatia (4.13 g each), Bombyx Batryticatus and Cimicifugae Rhizoma (2.891 g each), Bupleuri Radix and Platycodonis Radix (8.26 g each). These medicines were grounded to fine powder. One dose, including 20.65 g of the powder, was mixed with 600 mL of water and decocted to 300 mL. After abandoning slag, the medicine should be taken warm frequently. In the formula, Bombyx Batryticatus is stir-fired. With the effect of dispersing wind and clearing heat, removing stagnation and dissipating mass, the formula is specialized in swollen-head epidemic, pestilence, red and swelling head, face, and neck, dry mouth and tongue, as well as other diseases resulting from toxic heat stagnated in the upper jiao. The formula is widely used in treating diseases involving the respiratory, dermal, ophthalmologic, otolaryngologic, and nervous systems. The formula is most frequently used for respiratory diseases, with a wide range of symptoms including parotitis/mumps (66 times), followed by tonsillitis (28 times). In conclusion, the broadly applied formula has accurate efficacy and great development value.
4.Analysis of Risk Factors and Establishment of Prediction Model for Turbidity Toxicity Accumulation Syndrome in Patients with Chronic Atrophic Gastritis
Yican WANG ; Chenggong ZHAO ; Pengli DU ; Jie WANG ; Yuxi GUO ; Haiyan BAI ; Yongli HUO ; Xiaomeng LANG ; Zheng ZHI ; Bolin LI ; Jianping LIU ; Yanru CAI ; Jianming JIANG ; Qian YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):288-295
ObjectiveThis paper aims to explore the risk factors for chronic atrophic gastritis (CAG) with turbidity toxin accumulation syndrome and establish a prediction model. MethodsClinical data of 180 patients with CAG who participated in the "clinical study of Xianglian Huazhuo Particles blocking CAG cancer transformation" of Hebei Sheng Zhong Yi Yuan from July 2021 to March 2022 were collected. After confounding factors were controlled by propensity score matching, patients were divided into a training set (namely dev) and a validation set (namely vad) in a seven to three ratio. The risk factors for CAG with turbidity toxin accumulation syndrome in the training set were investigated by using univariate Logistic regression analysis and least absolute shrinkage and selection operator (namely Lasso) regression algorithms. Subsequently, a model, named model 1se, was developed by using the training set data to predict the risk factors for CAG with turbidity toxin accumulation syndrome. The accuracy of the prediction model was assessed by using various methods, including the receiver operating characteristic (ROC) curve, Hosmer-Lemeshow test (H-L), calibration plot, and decision curve analysis (DCA). ResultsAge, body mass index (BMI), family history of cancer, job and life satisfaction, yellow and greasy fur with slippery pulse, and heavy body sensation were independent risk factors of the model. The prediction model showed excellent predictive value for both the training and validation sets. ConclusionThe established prediction model for CAG with turbidity toxin accumulation syndrome has high discrimination and excellent calibration, which could provide an excellent clinical basis for disease diagnosis and individualized treatment of patients.
5.Investigating Effect of Xianglian Huazhuo Prescription on Cell Cycle and Proliferation in Rats with Chronic Atrophic Gastritis Through TGF-β1/Smads Signaling Pathway
Yican WANG ; Jie WANG ; Yirui CHENG ; Xiaojing LI ; Yibin MA ; Qiuhua LIU ; Ziwei LIU ; Yuxi GUO ; Pengli DU ; Yanru CAI ; Yao DU ; Zheng ZHI ; Bolin LI ; Qian YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):128-136
ObjectiveTo explore the potential mechanism of Xianglian Huazhuo prescription (XLHZ) in treating chronic atrophic gastritis (CAG) by regulating cell cycle and inhibiting proliferation, using bioinformatics technology and animal experiments. MethodsDifferential expressed genes (DEGs) related to CAG were screened using GEO database and GEO2R tool. Weighted gene co-expression network analysis (WGCNA) was employed to search for hub genes of CAG. These hub genes were intersected with cell cycle proliferation based on GeneCards database. Eenrichment analysis of the intersecting genes was performed to obtain signaling pathways and biological processes related to CAG. Protein protein interaction (PPI) analysis of genes was conducted using the Protein Interaction Platform (STRING) database to search the super hub gene (hub 2.0), and animal experiments were conducted for further validation. Fourteen of 70 male Wistar rats were randomly selected as the normal group, and the remaining 56 rats were prepared by the combined modeling method of "starvation disorder+N-methyl-N-nitro-N-nitrosoguanidine (MNNG) + sodium salicylate". The successfully modeled rats were randomly divided into the model group, XLHZ-H, XLHZ-M, and XLHZ-L groups (36, 18, 9 g·kg-1, respectively), and Morodan group (1.4 g·kg-1). Each group was given corresponding intervention for 60 days. Hematoxylin-eosin (HE) staining was used to observe the histopathological changes of gastric mucosa in rats. The ultrastructure of gastric mucosal tissue cells was observed by transmission electron microscopy. The relative expression levels of TGF-β1, Smad2 and Smad3 proteins, S/G2/M phase marker geminin and proliferation marker MCM2 were detected by Western blot in gastric mucosal tissue, and Spearman correlation analysis was performed. ResultsA total of 15 hub 2.0 genes were identified, including TGF-β1, suggesting the involvement of the TGF-β1 signaling pathway in the CAG pathogenesis. Compared with the normal group, the expressions of TGF-β1, Smad2, geminin and MCM2 proteins in the gastric mucosa tissue of the model group were increased (P<0.05), and the expression of Smad3 protein was decreased (P<0.05). Compared with the model group, the expressions of TGF-β1 and geminin in the gastric mucosa were decreased in the drug groups (P<0.05). The XLHZ-M group, XLHZ-H group and Morodan group had significantly decreased protein expression of Smad2 and MCM2 (P<0.05). The protein expression of Smad3 was significantly increased in XLHZ-M, XLHZ-H, and Morodan groups (P<0.05). Spearman correlation analysis showed that Smad3 was negatively correlated with other indicators, and positively correlated with other indicators (P<0.01). ConclusionXLHZ may inhibit TGF-β1/Smads signaling pathway, regulate cell cycle, and inhibit proliferation in the treatment of CAG.
6.Investigating Effect of Xianglian Huazhuo Prescription on Cell Cycle and Proliferation in Rats with Chronic Atrophic Gastritis Through TGF-β1/Smads Signaling Pathway
Yican WANG ; Jie WANG ; Yirui CHENG ; Xiaojing LI ; Yibin MA ; Qiuhua LIU ; Ziwei LIU ; Yuxi GUO ; Pengli DU ; Yanru CAI ; Yao DU ; Zheng ZHI ; Bolin LI ; Qian YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):128-136
ObjectiveTo explore the potential mechanism of Xianglian Huazhuo prescription (XLHZ) in treating chronic atrophic gastritis (CAG) by regulating cell cycle and inhibiting proliferation, using bioinformatics technology and animal experiments. MethodsDifferential expressed genes (DEGs) related to CAG were screened using GEO database and GEO2R tool. Weighted gene co-expression network analysis (WGCNA) was employed to search for hub genes of CAG. These hub genes were intersected with cell cycle proliferation based on GeneCards database. Eenrichment analysis of the intersecting genes was performed to obtain signaling pathways and biological processes related to CAG. Protein protein interaction (PPI) analysis of genes was conducted using the Protein Interaction Platform (STRING) database to search the super hub gene (hub 2.0), and animal experiments were conducted for further validation. Fourteen of 70 male Wistar rats were randomly selected as the normal group, and the remaining 56 rats were prepared by the combined modeling method of "starvation disorder+N-methyl-N-nitro-N-nitrosoguanidine (MNNG) + sodium salicylate". The successfully modeled rats were randomly divided into the model group, XLHZ-H, XLHZ-M, and XLHZ-L groups (36, 18, 9 g·kg-1, respectively), and Morodan group (1.4 g·kg-1). Each group was given corresponding intervention for 60 days. Hematoxylin-eosin (HE) staining was used to observe the histopathological changes of gastric mucosa in rats. The ultrastructure of gastric mucosal tissue cells was observed by transmission electron microscopy. The relative expression levels of TGF-β1, Smad2 and Smad3 proteins, S/G2/M phase marker geminin and proliferation marker MCM2 were detected by Western blot in gastric mucosal tissue, and Spearman correlation analysis was performed. ResultsA total of 15 hub 2.0 genes were identified, including TGF-β1, suggesting the involvement of the TGF-β1 signaling pathway in the CAG pathogenesis. Compared with the normal group, the expressions of TGF-β1, Smad2, geminin and MCM2 proteins in the gastric mucosa tissue of the model group were increased (P<0.05), and the expression of Smad3 protein was decreased (P<0.05). Compared with the model group, the expressions of TGF-β1 and geminin in the gastric mucosa were decreased in the drug groups (P<0.05). The XLHZ-M group, XLHZ-H group and Morodan group had significantly decreased protein expression of Smad2 and MCM2 (P<0.05). The protein expression of Smad3 was significantly increased in XLHZ-M, XLHZ-H, and Morodan groups (P<0.05). Spearman correlation analysis showed that Smad3 was negatively correlated with other indicators, and positively correlated with other indicators (P<0.01). ConclusionXLHZ may inhibit TGF-β1/Smads signaling pathway, regulate cell cycle, and inhibit proliferation in the treatment of CAG.
7.Level of vitamin D in children with cholestatic liver disease and its clinical features
Ziyun GUO ; Lina DU ; Xiaoxuan XIE ; Yan YANG
Journal of Clinical Hepatology 2025;41(1):99-103
ObjectiveTo investigate vitamin D level in children with cholestatic liver disease, and to provide a theoretical basis for vitamin D supplementation therapy in children with this disease. MethodsA total of 116 children with cholestatic liver disease who attended Department of Traditional Chinese Medicine, Beijing Children’s Hospital, Capital Medical University, for the first time from January 2022 to January 2024 were enrolled and divided into groups for comparison based on sex, age, vitamin D supplementation dose, course of the disease, and etiology. The data on the serum level of 25-hydroxyvitamin D (25-OH-D) and related biochemical parameters were collected to assess the correlation between vitamin D level and biochemical parameters. The chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups, and the Spearman rank correlation test was used for correlation analysis. ResultsAmong the 116 children, 76 (65.5%) had vitamin D deficiency or insufficiency. The children with vitamin D deficiency or insufficiency accounted for 65.7% (46/70) among boys and 65.2% (30/46) among girls, with no significant difference between boys and girls (χ2=0.003, P=0.956). The children with vitamin D deficiency or insufficiency accounted for 83.3% (25/30) among the children who had never received vitamin D supplementation, 58.7% (27/46) among the children with a daily supplementation dose of 500 IU, 64.3% (18/28) among the children with a daily supplementation dose of 700 IU, and 50.0% (6/12) among the children with a daily supplementation dose of>700 IU, and there was no significant difference between these groups (χ2=6.460, P=0.091). Comparison between the groups with different etiologies showed that the children with vitamin D deficiency or insufficiency accounted for 57.7% (15/26) in the infectious disease group, 66.7% (10/15) in the inherited metabolic disease group, 66.7% (6/9) in the drug-induced liver injury group, 100.0% (8/8) in the group with abnormal structure of the biliary system, and 63.8% (37/58) in the group with unknown etiology, and there was no significant difference between these groups (χ2=5.304, P=0.252). Comparison between the groups with different courses of the disease showed that the children with vitamin D deficiency or insufficiency accounted for 78.4% (29/37) in the<1 month group, 54.3% (25/46) in the 1 — 3 months group, 53.3% (8/15) in the 3 — 6 months group, and 77.8% (14/18) in the>6 months group, with no significant difference between these groups (χ2=7.432, P=0.059). Comparison between different age groups showed that compared with the infant group, the children group had a significantly higher proportion of children with vitamin D deficiency or insufficiency (χ2=9.504, P=0.018). The correlation analysis showed that serum aspartate aminotransferase and alanine aminotransferase had no significant correlation with 25-OH-D (P>0.05); serum alkaline phosphatase (ALP) (r=-0.286, P=0.002), gamma-glutamyl transpeptidase (GGT) (r=-0.248, P=0.007), total bilirubin (TBil) (r=-0.353, P<0.001), direct bilirubin (DBil) (r=-0.299, P=0.001), and total bile acid (r=-0.236, P=0.011) were negatively correlated with 25-OH-D, while serum calcium (r=0.263, P=0.004) and phosphorus (r=0.385, P<0.001) were positively correlated with 25-OH-D. ConclusionMost children with cholestatic liver disease have vitamin D deficiency or insufficiency, and the increase in serum ALP, GGT, TBil, DBil or total bile acid and the reduction in calcium or phosphorus may suggest vitamin D deficiency or insufficiency.
8.Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method
Yuan LI ; Hanqian DU ; Jiashan LI ; Li GUO ; Zehui LI ; Na LIN ; Ying XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):53-62
ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 (CCK-8) assay was employed to measure the viability of human ovarian granulosa cells (KGN cells) treated with Tusizi prescription extracts 1-11, and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM). The common targets shared by Tusizi prescription and diminished ovarian reserve (DOR) were selected and imported into search tool for the retrieval of interacting genes/proteins (STRING) to construct a protein-protein interaction (PPI) network and into gene function annotation database (DAVID) for gene ontology (GO) analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, terminal-deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL), and enzyme-linked immunosorbent assay (ELISA) were employed to examine the proliferation, apoptosis, and estradiol (E2) secretion of KGN cells treated with the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design. On this basis, the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes, primarily involving cellular responses to exogenous compound stimuli, negative regulation of apoptotic process, and positive regulation of cell proliferation. It identified 84 cellular components, including cell membrane, mitochondria, and neuronal cell body, as well as 144 molecular functions such as enzyme binding, estrogen response element binding, and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen, Lycii Fructus, Dioscoreae Rhizoma, Poria, and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14, the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group, the hyperoside group demonstrated increased viability of ovarian germline stem cells (P<0.01). The CCK-8, EdU, and ELISA results showed that compared with the normal group, the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells (P<0.05, P<0.01). ELISA results showed that compared with the normal group, the water extract 11 of Tusizi prescription promoted the E2 secretion of KGN cells (P<0.05), while the optimal prescription screened by uniform design had no significant effect on the E2 secretion. ConclusionBoth the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14) can improve the ovarian reserve function, and the former has better effect. Tusizi prescription can modulate biological processes (such as cell proliferation and apoptosis) and molecular functions (such as enzyme binding and estrogen response element binding) through active components like hyperoside to promote the proliferation and E2 secretion and inhibit the apoptosis of KGN cells, thereby protecting the ovarian reserve function.
9.Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method
Yuan LI ; Hanqian DU ; Jiashan LI ; Li GUO ; Zehui LI ; Na LIN ; Ying XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):53-62
ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 (CCK-8) assay was employed to measure the viability of human ovarian granulosa cells (KGN cells) treated with Tusizi prescription extracts 1-11, and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM). The common targets shared by Tusizi prescription and diminished ovarian reserve (DOR) were selected and imported into search tool for the retrieval of interacting genes/proteins (STRING) to construct a protein-protein interaction (PPI) network and into gene function annotation database (DAVID) for gene ontology (GO) analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, terminal-deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL), and enzyme-linked immunosorbent assay (ELISA) were employed to examine the proliferation, apoptosis, and estradiol (E2) secretion of KGN cells treated with the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design. On this basis, the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes, primarily involving cellular responses to exogenous compound stimuli, negative regulation of apoptotic process, and positive regulation of cell proliferation. It identified 84 cellular components, including cell membrane, mitochondria, and neuronal cell body, as well as 144 molecular functions such as enzyme binding, estrogen response element binding, and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen, Lycii Fructus, Dioscoreae Rhizoma, Poria, and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14, the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group, the hyperoside group demonstrated increased viability of ovarian germline stem cells (P<0.01). The CCK-8, EdU, and ELISA results showed that compared with the normal group, the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells (P<0.05, P<0.01). ELISA results showed that compared with the normal group, the water extract 11 of Tusizi prescription promoted the E2 secretion of KGN cells (P<0.05), while the optimal prescription screened by uniform design had no significant effect on the E2 secretion. ConclusionBoth the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14) can improve the ovarian reserve function, and the former has better effect. Tusizi prescription can modulate biological processes (such as cell proliferation and apoptosis) and molecular functions (such as enzyme binding and estrogen response element binding) through active components like hyperoside to promote the proliferation and E2 secretion and inhibit the apoptosis of KGN cells, thereby protecting the ovarian reserve function.
10.Intervention Effect of Suanzaoren Tang on Depression Model Rats Based on JNK/c-Myc/p53 Pathway
Shuailin DU ; Zhicheng HAO ; Ce ZHANG ; Jiyuan GUO ; Xusheng TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):12-19
ObjectiveTo investigate the intervention effects of Suanzaoren Tang on depression model rats induced by isolation combined with chronic unpredictable mild stress (CUMS), and to examine its influence on the c-Jun N-terminal kinase (JNK)/proto-oncogene protein (c-Myc)/tumor suppressor protein 53 (p53) signaling pathway, thereby revealing its potential functional mechanism. MethodsA total of 72 male SD rats were randomly divided into six groups using a strict random number table: blank group, model group, fluoxetine group (3.6 mg·kg-1), and high-, medium-, and low-dose Suanzaoren Tang groups (10, 5, 2.5 g·kg-1),with 12 rats in each group. A depression model was established using isolation combined with CUMS. Fluoxetine and different doses of Suanzaoren Tang were administered continuously for 28 days. Behavioral indicators such as sucrose water consumption and open field test scores were recorded. Western blot and immunohistochemistry (IHC) were employed to analyze the expression of key proteins in the JNK/c-Myc/p53 signaling pathway, and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to evaluate the number of apoptotic cells in the hippocampus. ResultsCompared with the blank group, the model group exhibited a significantly reduced sucrose preference index (P<0.01), a lower total score of horizontal and vertical movements in the open field test (P<0.01), significantly increased expression of JNK, c-Myc, and p53 proteins in the hippocampus (P<0.01), and a higher number of TUNEL-positive cells in the hippocampus (P<0.01). Compared with the model group, the sucrose preference index and the total score of horizontal and vertical movements in the open field test significantly increased in the high- and medium-dose Suanzaoren Tang groups and the fluoxetine group (P<0.05, P<0.01). The expression of JNK, c-Myc, and p53 proteins significantly decreased in all Suanzaoren Tang groups (high, medium, and low doses) and the fluoxetine group (P<0.05, P<0.01). The number of TUNEL-positive cells in the hippocampus also significantly decreased in these groups (P<0.01). ConclusionSuanzaoren Tang can regulate the expression of JNK/c-Myc/p53 proteins in the hippocampus of depression model rats, and its antidepressant mechanism may be related to its protective effect on hippocampal neurons.

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