1.Comparative Study on Effect of Jingui Shenqiwan and Liuwei Dihuangwan on Reproductive Ability and Brain Function of Normal Mice
Hong SUN ; Fan LEI ; Chenggong LI ; Rui LUO ; Shixian HU ; Bin REN ; Juan HAO ; Yi DING ; Lijun DU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):1-14
ObjectiveTo explore the effects of Jingui Shenqiwan (JSW) and Liuwei Dihuangwan (LDW) on the reproductive ability and brain function of normal mice and compare the actions of the two medications. MethodsSeven groups of female and male mice were divided at a ratio of 2∶1. Except for the control group, the other six groups were as follows: a group of both males and females receiving JSW (3.0 g·kg-1), a group of both males and females receiving LDW (4.5 g·kg-1), a group of males receiving water and females receiving JSW, a group of males receiving water while females receiving LDW, a group of females receiving water while males receiving JSW, and a group of females receiving water while males receiving LDW. Each group was administered the drug for 14 days and then caged together at a 2∶1 (female∶male) ratio to detect the number of pregnant mice and calculate the pregnancy rate. Pregnant mice continued receiving the drug until they naturally gave birth, which was followed by the observation of newborn mice, calculation of their average number, and the measurement of the offspring's preference for sugar water and neonatal recognition index. At the end of the experiment, the weights of the thymus and spleen were measured to calculate the organ coefficients, and mRNA or protein expression was analyzed in the brain and testes or ovaries. A 1% sucrose solution was used to examine the euphoria of their brain reward systems, while novel object recognition test (NOR) was applied to assess their memory capabilities. mRNA expression was detected using real-time quantitative polymerase chain reaction (Real-time PCR) assay, and protein expression was analyzed with Western blot. ResultsCompared with the control group, oral administration of JSW to both male and female mice for 14 days significantly increased the pregnancy rate of female mice on day 2 after being caged together (P<0.05), while LDW showed a trend but no statistical significance. Additionally, compared with the control group, JSW could upregulate the gene expression of gonadotropin-releasing hormone (GnRH) in the thalamus, as well as reproductive stem cell factor (SCF) and tyrosine kinase receptor (c-Kit) in the testes and reproductive stem cell marker mouse vasa homologue (MVH) in the ovaries, upregulate the expression of proteins influencing neuronal functional activity, such as brain-derived neurotrophic factor (BDNF), in hippocampal neurons (P<0.05), and enhance sucrose preference in male mice (P<0.05). Compared with the control group, JSW significantly increased sucrose preference and novel object recognition index in offspring mice (P<0.05), which was related to the upregulation of hippocampal dopamine D1 receptor (D1R) and N-methyl-D-aspartate receptor (Nmdar) gene expression. Compared with the control group, both JSW and LDW could upregulate the protein expression of glucocorticoid receptor (GR), BDNF, and tyrosine kinase receptor B (TrkB) in the hippocampus of offspring mice (P<0.05). ConclusionJSW significantly enhances the reproductive ability of normal mice, which is not only related to the release of gonadotropin but also associated with its regulation of brain function. Additionally, JSW has a certain regulatory effect on the brain function of the offspring mice.
2.Quality Evaluation of Naomaili Granules Based on Multi-component Content Determination and Fingerprint and Screening of Its Anti-neuroinflammatory Substance Basis
Ya WANG ; Yanan KANG ; Bo LIU ; Zimo WANG ; Xuan ZHANG ; Wei LAN ; Wen ZHANG ; Lu YANG ; Yi SUN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):170-178
ObjectiveTo establish an ultra-performance liquid fingerprint and multi-components determination method for Naomaili granules. To evaluate the quality of different batches by chemometrics, and the anti-neuroinflammatory effects of water extract and main components of Naomaili granules were tested in vitro. MethodsThe similarity and common peaks of 27 batches of Naomaili granules were evaluated by using Ultra performance liquid chromatography (UPLC) fingerprint detection. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technology was used to determine the content of the index components in Naomaili granules and to evaluate the quality of different batches of Naomaili granules by chemometrics. LPS-induced BV-2 cell inflammation model was used to investigate the anti-neuroinflammatory effects of the water extract and main components of Naomaili granules. ResultsThe similarity of fingerprints of 27 batches of samples was > 0.90. A total of 32 common peaks were calibrated, and 23 of them were identified and assigned. In 27 batches of Naomaili granules, the mass fractions of 14 components that were stachydrine hydrochloride, leonurine hydrochloride, calycosin-7-O-glucoside, calycosin,tanshinoneⅠ, cryptotanshinone, tanshinoneⅡA, ginsenoside Rb1, notoginsenoside R1, ginsenoside Rg1, paeoniflorin, albiflorin, lactiflorin, and salvianolic acid B were found to be 2.902-3.498, 0.233-0.343, 0.111-0.301, 0.07-0.152, 0.136-0.228, 0.195-0.390, 0.324-0.482, 1.056-1.435, 0.271-0.397, 1.318-1.649, 3.038-4.059, 2.263-3.455, 0.152-0.232, 2.931-3.991 mg∙g-1, respectively. Multivariate statistical analysis showed that paeoniflorin, ginsenoside Rg1, ginsenoside Rb1 and staphylline hydrochloride were quality difference markers to control the stability of the preparation. The results of bioactive experiment showed that the water extract of Naomaili granules and the eight main components with high content in the prescription had a dose-dependent inhibitory effect on the release of NO in the cell supernatant. Among them, salvianolic acid B and ginsenoside Rb1 had strong anti-inflammatory activity, with IC50 values of (36.11±0.15) mg∙L-1 and (27.24±0.54) mg∙L-1, respectively. ConclusionThe quality evaluation method of Naomaili granules established in this study was accurate and reproducible. Four quality difference markers were screened out, and eight key pharmacodynamic substances of Naomaili granules against neuroinflammation were screened out by in vitro cell experiments.
3.Molecular Mechanism of Astragali Radix and Hedyotis diffusa in Regulating LINC01134-CTCF-p21 Axis to Inhibit Lung Adenocarcinoma Proliferation
Haipeng SUN ; He ZHUANG ; Xue LIU ; Siyuan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):131-138
ObjectiveTo explore the interaction and competitive binding of Homo sapiens long intergenic non-protein-coding RNA 1134 (LINC01134) to CCCTC-binding factor CTCF, affecting the transcription of cyclin-dependent kinase inhibitor (p21) and influencing the proliferation of A549 cells, in order to investigate the possible mechanism of Astragali Radix and Hedyotis diffusa (A-H) in inhibiting A549 proliferation by regulating this axis. MethodsRNA-binding protein immunoprecipitation (RIP) assays were conducted to examine the interaction between LINC01134 and CTCF, and chromatin immunoprecipitation (ChIP) assays were used to study the effect of LINC01134 overexpression on the interaction between CTCF and p21. Stable A549 cell lines (oe-NC and oe-LINC01134) were established using lentiviral transfection, and each group was treated with 10% A-H drug-containing serum. Real-time PCR and Western blot analyses were performed to detect the effects of A-H on the expression of LINC01134, CTCF, and p21 in A549 cells. Cell counting kit-8 (CCK-8) and colony formation assays were used to assess the effects of A-H on A549 cell proliferation via LINC01134. Flow cytometry was employed to evaluate the effects of A-H on the A549 cell cycle through LINC01134, and Western blot was used to detect changes in cell cycle proteins. ResultsCompared with the IgG group, the oe-CTCF group showed a significantly increased abundance of LINC01134 aggregates (P0.01). Compared with the oe-Vector group, p21 abundance in CTCF complexes was significantly reduced in the oe-LINC01134 group (P0.01). Compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed the expression of LINC01134 and p21 (P0.05), but had no significant regulatory effect on CTCF. Compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed cell viability at 72 h (P0.05), inhibited malignant proliferation (P0.05), and reversed the proportions of cells in the G0/G1 and S phases (P0.01). Furthermore, compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed the expression of Cyclin D1, CDK4, Cyclin E, CDK2, phosphorylated retinoblastoma protein (p-Rb), and E2F transcription factor 3 (E2F3) (P0.01). ConclusionA-H regulates the LINC01134-CTCF-p21 axis to block the G1/S phase transition of A549 cell cycle, accelerate cellular senescence, and inhibit malignant proliferation.
4.Analysis of Animal Models of Retinal Vein Occlusion Based on Clinical Manifestations of Traditional Chinese and Western Medicine
Xiaoyu LI ; Lina LIANG ; Xiaofeng HAO ; Menglu MIAO ; Mei SUN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):204-209
ObjectiveRetinal vein occlusion (RVO) is the second most common vascular disease leading to vision loss. Since its pathogenesis remains unclear, current Western medical treatments primarily target complications such as macular edema and neovascularization. The main therapeutic approaches include intravitreal injections of anti-vascular endothelial growth factor (VEGF) agents or corticosteroids, laser photocoagulation, and pars plana vitrectomy. However, these treatments cannot fully reverse disease progression or structural damage. Traditional Chinese medicine (TCM) has unique advantages in the clinical diagnosis and treatment of RVO, and integrated Chinese and Western medicine approaches may offer better clinical outcomes. This study, based on the clinical manifestations of RVO, systematically reviews the existing literature and evaluates the alignment of current RVO animal models with clinical manifestations. The aim is to identify the characteristics and limitations of existing models and provide recommendations and prospects for developing RVO animal models featuring the combination of disease and syndrome. MethodsDatabases including CNKI, Wanfang Data, PubMed, and Web of Science were searched with the keywords of "retinal vein occlusion" and "animal model". Model characteristics were assessed based on the diagnostic criteria for diseases and syndromes in both TCM and Western medicine. The alignment of each model with clinical manifestations was analyzed and evaluated. ResultsThe available RVO models were primarily established via methods such as laser photocoagulation, photodynamic therapy, diathermy, intravitreal drug injection, and mechanical modeling. These models demonstrated moderate overall alignment with clinical manifestations, mainly reflecting disease characteristics. However, they generally lack representation of TCM syndrome features. ConclusionExisting RVO models are predominantly based on Western medicine and lack TCM syndrome features. Western medical treatments for RVO have certain limitations, while syndrome differentiation and treatment in TCM offer potential advantages. Future research should focus on developing disease-syndrome integrated animal models that incorporate both pathological features and TCM syndrome characteristics. This approach will enhance the design of RVO models and facilitate both basic and clinical research, which make it a scientifically valuable and necessary endeavor.
5.Exploring Pathogenesis and Treatment Principles of Chronic Obstructive Pulmonary Disease Based on Spleen-mitochondria Correlation
Shiyi WANG ; Miao YU ; Xinyao HE ; Zi WANG ; Haijun LUAN ; Yibo SUN ; Haotong WANG ; Linlin WANG ; Lijian PANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):258-264
According to the Qi-blood-body fluid theory and the association between the spleen in visceral manifestation theory of traditional Chinese medicine (TCM) and mitochondria in modern cellular biology, it is proposed that the role of the spleen in generating and transforming Qi and blood is analogous to the energy-producing function of mitochondria—both serving as fundamental power sources for vital activities of the human body. The spleen governs transportation and transformation, playing a critical role in energy metabolism and the digestion and absorption of nutrients. Similarly, mitochondria are vital for maintaining physiological functions such as cellular energy supply, cell survival, and overall human metabolism. Furthermore, spleen deficiency is closely linked to mitochondrial dysfunction. Accordingly, mitochondrial energy conversion and substance metabolism are regarded as the microscopic essence of the spleen's function in transportation and transformation. Spleen deficiency and mitochondrial dysfunction contribute to the formation of pathological products such as phlegm-turbidity and blood stasis. This aligns with the pathogenesis of chronic obstructive pulmonary disease (COPD), with Qi deficiency as the root cause and phlegm-turbidity and blood stasis as the manifestations. Therefore, the integrative treatment of COPD should follow the therapeutic principle of invigorating the spleen and reinforcing healthy Qi, while also resolving phlegm and removing blood stasis to address both root cause and manifestations. This approach can improve the mitochondrial function, regulate energy metabolism, and reduce oxidative stress levels to alleviate COPD symptoms, slow down disease progression, and improve prognosis. By integrating the holistic concept of TCM with molecular mechanisms of modern medicine, this paper explores the pathogenesis and therapeutic principles of COPD from the spleen-mitochondria correlation. It not only provides a new direction for the modern development of TCM and the integration of Chinese and Western medicine but also offers a theoretical foundation for the integrated treatment of chronic, complex age-related diseases.
6.Xuefu Zhuyutang in Malignant Tumor Disease: A Review
Jiaqi JI ; Xiaoqing HU ; Yihan ZHAO ; Xuhang SUN ; Dandan WEI ; Junwen PEI ; Shiqing JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):321-330
Cancer has become a significant global public health issue, severely impacting public health and societal development. Despite advances in tumor treatment methods in recent years and a gradual decline in cancer mortality rates, drug-related adverse reactions and drug resistance remain substantial challenges. Traditional Chinese medicine (TCM) has demonstrated significant clinical efficacy in cancer treatment and small side effects, making it widely applied in the field of oncology. Xuefu Zhuyutang, derived from Yilin Gaicuo, is known for its abilities to invigorate blood circulation, dispel blood stasis, promote Qi flow, and alleviate pain. It was specifically formulated by the esteemed WANG Qingren of the Qing dynasty for the "blood stasis syndrome in the blood mansion" and is commonly used to treat Qi stagnation and blood stasis syndrome. Clinical studies have shown that Xuefu Zhuyutang, when combined with conventional Western medications, produces significant effects in the treatment of malignant tumors such as liver cancer, lung cancer, and cervical cancer. It substantially reduces the incidence of adverse reactions following Western treatments, including radiation esophagitis, radiation encephalopathy, radiation-induced oral mucositis, and edema. Additionally, it alleviates cancer-related pain and fever, blood hypercoagulability, and associated complications such as depression and anxiety, and also mitigates chemotherapy-induced side effects like hand-foot syndrome. Basic research has demonstrated its potential anti-tumor mechanisms, including the inhibition of Wnt/β-catenin signaling pathway activation, suppression of mitogen-activated protein kinase (MAPK) pathway activation, and anti-tumor angiogenesis. Pharmacological studies have revealed that its active components inhibit tumor cell proliferation and migration, induce tumor cell apoptosis, suppress tumor angiogenesis, enhance the cytotoxicity of natural killer cells against tumors, improve the tumor microenvironment, and regulate immune function. This paper reviewed the latest research progress on Xuefu Zhuyutang in the treatment of malignant tumors from four aspects: theoretical exploration, clinical studies, mechanisms of action, and pharmacological basis, aiming to provide insights and methods for the clinical diagnosis and treatment of malignant tumors.
7.Molecular Mechanism of Astragali Radix and Hedyotis diffusa in Regulating LINC01134-CTCF-p21 Axis to Inhibit Lung Adenocarcinoma Proliferation
Haipeng SUN ; He ZHUANG ; Xue LIU ; Siyuan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):131-138
ObjectiveTo explore the interaction and competitive binding of Homo sapiens long intergenic non-protein-coding RNA 1134 (LINC01134) to CCCTC-binding factor CTCF, affecting the transcription of cyclin-dependent kinase inhibitor (p21) and influencing the proliferation of A549 cells, in order to investigate the possible mechanism of Astragali Radix and Hedyotis diffusa (A-H) in inhibiting A549 proliferation by regulating this axis. MethodsRNA-binding protein immunoprecipitation (RIP) assays were conducted to examine the interaction between LINC01134 and CTCF, and chromatin immunoprecipitation (ChIP) assays were used to study the effect of LINC01134 overexpression on the interaction between CTCF and p21. Stable A549 cell lines (oe-NC and oe-LINC01134) were established using lentiviral transfection, and each group was treated with 10% A-H drug-containing serum. Real-time PCR and Western blot analyses were performed to detect the effects of A-H on the expression of LINC01134, CTCF, and p21 in A549 cells. Cell counting kit-8 (CCK-8) and colony formation assays were used to assess the effects of A-H on A549 cell proliferation via LINC01134. Flow cytometry was employed to evaluate the effects of A-H on the A549 cell cycle through LINC01134, and Western blot was used to detect changes in cell cycle proteins. ResultsCompared with the IgG group, the oe-CTCF group showed a significantly increased abundance of LINC01134 aggregates (P0.01). Compared with the oe-Vector group, p21 abundance in CTCF complexes was significantly reduced in the oe-LINC01134 group (P0.01). Compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed the expression of LINC01134 and p21 (P0.05), but had no significant regulatory effect on CTCF. Compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed cell viability at 72 h (P0.05), inhibited malignant proliferation (P0.05), and reversed the proportions of cells in the G0/G1 and S phases (P0.01). Furthermore, compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed the expression of Cyclin D1, CDK4, Cyclin E, CDK2, phosphorylated retinoblastoma protein (p-Rb), and E2F transcription factor 3 (E2F3) (P0.01). ConclusionA-H regulates the LINC01134-CTCF-p21 axis to block the G1/S phase transition of A549 cell cycle, accelerate cellular senescence, and inhibit malignant proliferation.
8.Analysis of Animal Models of Retinal Vein Occlusion Based on Clinical Manifestations of Traditional Chinese and Western Medicine
Xiaoyu LI ; Lina LIANG ; Xiaofeng HAO ; Menglu MIAO ; Mei SUN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):204-209
ObjectiveRetinal vein occlusion (RVO) is the second most common vascular disease leading to vision loss. Since its pathogenesis remains unclear, current Western medical treatments primarily target complications such as macular edema and neovascularization. The main therapeutic approaches include intravitreal injections of anti-vascular endothelial growth factor (VEGF) agents or corticosteroids, laser photocoagulation, and pars plana vitrectomy. However, these treatments cannot fully reverse disease progression or structural damage. Traditional Chinese medicine (TCM) has unique advantages in the clinical diagnosis and treatment of RVO, and integrated Chinese and Western medicine approaches may offer better clinical outcomes. This study, based on the clinical manifestations of RVO, systematically reviews the existing literature and evaluates the alignment of current RVO animal models with clinical manifestations. The aim is to identify the characteristics and limitations of existing models and provide recommendations and prospects for developing RVO animal models featuring the combination of disease and syndrome. MethodsDatabases including CNKI, Wanfang Data, PubMed, and Web of Science were searched with the keywords of "retinal vein occlusion" and "animal model". Model characteristics were assessed based on the diagnostic criteria for diseases and syndromes in both TCM and Western medicine. The alignment of each model with clinical manifestations was analyzed and evaluated. ResultsThe available RVO models were primarily established via methods such as laser photocoagulation, photodynamic therapy, diathermy, intravitreal drug injection, and mechanical modeling. These models demonstrated moderate overall alignment with clinical manifestations, mainly reflecting disease characteristics. However, they generally lack representation of TCM syndrome features. ConclusionExisting RVO models are predominantly based on Western medicine and lack TCM syndrome features. Western medical treatments for RVO have certain limitations, while syndrome differentiation and treatment in TCM offer potential advantages. Future research should focus on developing disease-syndrome integrated animal models that incorporate both pathological features and TCM syndrome characteristics. This approach will enhance the design of RVO models and facilitate both basic and clinical research, which make it a scientifically valuable and necessary endeavor.
9.Exploring Pathogenesis and Treatment Principles of Chronic Obstructive Pulmonary Disease Based on Spleen-mitochondria Correlation
Shiyi WANG ; Miao YU ; Xinyao HE ; Zi WANG ; Haijun LUAN ; Yibo SUN ; Haotong WANG ; Linlin WANG ; Lijian PANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):258-264
According to the Qi-blood-body fluid theory and the association between the spleen in visceral manifestation theory of traditional Chinese medicine (TCM) and mitochondria in modern cellular biology, it is proposed that the role of the spleen in generating and transforming Qi and blood is analogous to the energy-producing function of mitochondria—both serving as fundamental power sources for vital activities of the human body. The spleen governs transportation and transformation, playing a critical role in energy metabolism and the digestion and absorption of nutrients. Similarly, mitochondria are vital for maintaining physiological functions such as cellular energy supply, cell survival, and overall human metabolism. Furthermore, spleen deficiency is closely linked to mitochondrial dysfunction. Accordingly, mitochondrial energy conversion and substance metabolism are regarded as the microscopic essence of the spleen's function in transportation and transformation. Spleen deficiency and mitochondrial dysfunction contribute to the formation of pathological products such as phlegm-turbidity and blood stasis. This aligns with the pathogenesis of chronic obstructive pulmonary disease (COPD), with Qi deficiency as the root cause and phlegm-turbidity and blood stasis as the manifestations. Therefore, the integrative treatment of COPD should follow the therapeutic principle of invigorating the spleen and reinforcing healthy Qi, while also resolving phlegm and removing blood stasis to address both root cause and manifestations. This approach can improve the mitochondrial function, regulate energy metabolism, and reduce oxidative stress levels to alleviate COPD symptoms, slow down disease progression, and improve prognosis. By integrating the holistic concept of TCM with molecular mechanisms of modern medicine, this paper explores the pathogenesis and therapeutic principles of COPD from the spleen-mitochondria correlation. It not only provides a new direction for the modern development of TCM and the integration of Chinese and Western medicine but also offers a theoretical foundation for the integrated treatment of chronic, complex age-related diseases.
10.Xuefu Zhuyutang in Malignant Tumor Disease: A Review
Jiaqi JI ; Xiaoqing HU ; Yihan ZHAO ; Xuhang SUN ; Dandan WEI ; Junwen PEI ; Shiqing JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):321-330
Cancer has become a significant global public health issue, severely impacting public health and societal development. Despite advances in tumor treatment methods in recent years and a gradual decline in cancer mortality rates, drug-related adverse reactions and drug resistance remain substantial challenges. Traditional Chinese medicine (TCM) has demonstrated significant clinical efficacy in cancer treatment and small side effects, making it widely applied in the field of oncology. Xuefu Zhuyutang, derived from Yilin Gaicuo, is known for its abilities to invigorate blood circulation, dispel blood stasis, promote Qi flow, and alleviate pain. It was specifically formulated by the esteemed WANG Qingren of the Qing dynasty for the "blood stasis syndrome in the blood mansion" and is commonly used to treat Qi stagnation and blood stasis syndrome. Clinical studies have shown that Xuefu Zhuyutang, when combined with conventional Western medications, produces significant effects in the treatment of malignant tumors such as liver cancer, lung cancer, and cervical cancer. It substantially reduces the incidence of adverse reactions following Western treatments, including radiation esophagitis, radiation encephalopathy, radiation-induced oral mucositis, and edema. Additionally, it alleviates cancer-related pain and fever, blood hypercoagulability, and associated complications such as depression and anxiety, and also mitigates chemotherapy-induced side effects like hand-foot syndrome. Basic research has demonstrated its potential anti-tumor mechanisms, including the inhibition of Wnt/β-catenin signaling pathway activation, suppression of mitogen-activated protein kinase (MAPK) pathway activation, and anti-tumor angiogenesis. Pharmacological studies have revealed that its active components inhibit tumor cell proliferation and migration, induce tumor cell apoptosis, suppress tumor angiogenesis, enhance the cytotoxicity of natural killer cells against tumors, improve the tumor microenvironment, and regulate immune function. This paper reviewed the latest research progress on Xuefu Zhuyutang in the treatment of malignant tumors from four aspects: theoretical exploration, clinical studies, mechanisms of action, and pharmacological basis, aiming to provide insights and methods for the clinical diagnosis and treatment of malignant tumors.

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