1.Mechanism of Yizhi Qingxin Prescription in Regulating PKA/CaN Pathway to Improve Cognitive Function in Alzheimer's Disease Model Mice
Xiaochen GUO ; Jiangang LIU ; Dandan SHI ; Ziqi NING ; Yaoyao ZHANG ; Fang LIU ; Meixia LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):97-108
ObjectiveTo explore the mechanism by which Yizhi Qingxin prescription improves mitochondrial dysfunction in Alzheimer's disease (AD) through regulating mitochondrial Ca2+ homeostasis and kinetic balance based on the protein kinase A (PKA)/calcineurin (CaN) signaling pathway. MethodsSixty three-month-old amyloid precursor protein (APP)/presenilin 1 (PS1) double transgenic mice were randomly divided into a model group, a donepezil group(0.65 mg·kg-1), a low-dose Yizhi Qingxin prescription group (YQF-L,2.6 g·kg-1), a medium-dose Yizhi Qingxin prescription group (YQF-M,5.2 g·kg-1), and a high-dose Yizhi Qingxin prescription group (YQF-H,10.4 g·kg-1), with 12 mice in each group. Twelve C57BL/6J mice with the same genetic background served as a normal group. Each treatment group received gavage administration daily, with the model and normal groups receiving equal volume of physiological saline. Intervention continued for 12 consecutive weeks. The learning and memory abilities of the mice were assessed using the novel object recognition (NOR) and Morris water maze (MWM) tests. Hematoxylin-eosin (HE)/Nissl staining was used to observe histopathological changes in the hippocampus. Transmission electron microscopy (TEM) was used to observe mitochondrial ultrastructure. Fluo-4 acetoxymethyl ester (Fluo-4 AM) Ca2+ probe was used to measure intracellular Ca2+ concentration in brain tissue. Western blot was used to determine the protein expression of PKA, CaN, sodium/calcium/lithium exchanger (NCLX), mitochondrial calcium uniporter (MCU), calmodulin (CaM), dynamin-related protein 1 (Drp1), and phosphorylated dynamin-related protein 1 (serine 637 site) [p-Drp1(S637)] in the hippocampus. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to measure the expression of PKA, CaN, CaM, NCLX, MCU, and Drp1 mRNAs. ResultsCompared with those in the normal group, the recognition index (RI) of the model group decreased (P0.01), and the number of crossings through the original platform area, the duration of stay in the target quadrant, and the distance were reduced (P0.01). The protein expression of PKA, NCLX, and p-DRP1 (ser637) significantly decreased (P0.05), and the mRNA expression of PKA and NCLX significantly decreased (P0.05). The escape latency (EL) was prolonged (P0.05), and the intracellular Ca2+ level significantly increased (P0.01). The protein expression of CaN, CaM, MCU, and Drp1, as well as the mRNA expression of CaN, MCU, and Drp1, significantly increased (P0.05). After intervention with Donepezil and Yizhi Qingxin prescription, compared with that in the model group, the RI of the treatment group significantly increased (P0.05), and the number of crossings through the platform and the duration of stay in the target quadrant significantly increased (P0.05). The protein expression of PKA, NCLX, and p-Drp1 (ser637) and the mRNA expression of PKA and NCLX significantly increased (P0.05). On the 4th and 5th days, the EL was shortened (P0.05), and the intracellular Ca2+ level decreased (P0.05). The protein expression of CaN, CaM, MCU, and Drp1 and the mRNA expression of CaN, MCU, and Drp1 significantly decreased (P0.05). ConclusionYizhi Qingxin prescription regulates the PKA/CaN pathway, upregulates the expression of PKA, NCLX, and p-Drp1 (ser637) proteins, reduces the expression of CaN, CaM, MCU, and Drp1 proteins, and regulates Ca2+ homeostasis and mitochondrial dynamic balance, thereby enhancing the spatial learning and memory abilities of AD mice.
2.Mechanism of Yizhi Qingxin Prescription in Regulating PKA/CaN Pathway to Improve Cognitive Function in Alzheimer's Disease Model Mice
Xiaochen GUO ; Jiangang LIU ; Dandan SHI ; Ziqi NING ; Yaoyao ZHANG ; Fang LIU ; Meixia LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):97-108
ObjectiveTo explore the mechanism by which Yizhi Qingxin prescription improves mitochondrial dysfunction in Alzheimer's disease (AD) through regulating mitochondrial Ca2+ homeostasis and kinetic balance based on the protein kinase A (PKA)/calcineurin (CaN) signaling pathway. MethodsSixty three-month-old amyloid precursor protein (APP)/presenilin 1 (PS1) double transgenic mice were randomly divided into a model group, a donepezil group(0.65 mg·kg-1), a low-dose Yizhi Qingxin prescription group (YQF-L,2.6 g·kg-1), a medium-dose Yizhi Qingxin prescription group (YQF-M,5.2 g·kg-1), and a high-dose Yizhi Qingxin prescription group (YQF-H,10.4 g·kg-1), with 12 mice in each group. Twelve C57BL/6J mice with the same genetic background served as a normal group. Each treatment group received gavage administration daily, with the model and normal groups receiving equal volume of physiological saline. Intervention continued for 12 consecutive weeks. The learning and memory abilities of the mice were assessed using the novel object recognition (NOR) and Morris water maze (MWM) tests. Hematoxylin-eosin (HE)/Nissl staining was used to observe histopathological changes in the hippocampus. Transmission electron microscopy (TEM) was used to observe mitochondrial ultrastructure. Fluo-4 acetoxymethyl ester (Fluo-4 AM) Ca2+ probe was used to measure intracellular Ca2+ concentration in brain tissue. Western blot was used to determine the protein expression of PKA, CaN, sodium/calcium/lithium exchanger (NCLX), mitochondrial calcium uniporter (MCU), calmodulin (CaM), dynamin-related protein 1 (Drp1), and phosphorylated dynamin-related protein 1 (serine 637 site) [p-Drp1(S637)] in the hippocampus. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to measure the expression of PKA, CaN, CaM, NCLX, MCU, and Drp1 mRNAs. ResultsCompared with those in the normal group, the recognition index (RI) of the model group decreased (P0.01), and the number of crossings through the original platform area, the duration of stay in the target quadrant, and the distance were reduced (P0.01). The protein expression of PKA, NCLX, and p-DRP1 (ser637) significantly decreased (P0.05), and the mRNA expression of PKA and NCLX significantly decreased (P0.05). The escape latency (EL) was prolonged (P0.05), and the intracellular Ca2+ level significantly increased (P0.01). The protein expression of CaN, CaM, MCU, and Drp1, as well as the mRNA expression of CaN, MCU, and Drp1, significantly increased (P0.05). After intervention with Donepezil and Yizhi Qingxin prescription, compared with that in the model group, the RI of the treatment group significantly increased (P0.05), and the number of crossings through the platform and the duration of stay in the target quadrant significantly increased (P0.05). The protein expression of PKA, NCLX, and p-Drp1 (ser637) and the mRNA expression of PKA and NCLX significantly increased (P0.05). On the 4th and 5th days, the EL was shortened (P0.05), and the intracellular Ca2+ level decreased (P0.05). The protein expression of CaN, CaM, MCU, and Drp1 and the mRNA expression of CaN, MCU, and Drp1 significantly decreased (P0.05). ConclusionYizhi Qingxin prescription regulates the PKA/CaN pathway, upregulates the expression of PKA, NCLX, and p-Drp1 (ser637) proteins, reduces the expression of CaN, CaM, MCU, and Drp1 proteins, and regulates Ca2+ homeostasis and mitochondrial dynamic balance, thereby enhancing the spatial learning and memory abilities of AD mice.
3.Clinical Efficacy of Gandou Fumu Decoction in Treating Hepatolenticular Degeneration with Liver Fibrosis of Liver-kidney Deficiency and Phlegm-blood Stasis Syndrome
Pingping YANG ; Meixia WANG ; Changchang CAO ; Zhuang TAO ; Jiang DU ; Yun XU ; Wenming YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):182-191
ObjectiveThis paper aims to evaluate the intervention effect of Gandou Fumu Decoction (GDFMD) in treating hepatolenticular degeneration with liver fibrosis of liver-kidney deficiency and phlegm-blood stasis syndrome, thereby providing evidence-based medical evidence for the treatment of Wilson's disease (WD)-related liver fibrosis with traditional Chinese medicine through clinical efficacy analysis. MethodsA total of 70 patients with WD-related liver fibrosis of liver-kidney deficiency and phlegm-blood stasis syndrome meeting the inclusion criteria were enrolled from Anhui Provincial Hospital of TCM from October 1, 2023, to October 1, 2024. Participants were divided into a control group and an observation group, with 35 cases in each group. The control group received conventional copper chelation therapy with sodium dimercaptopropanesulfonate (DMPS). On this basis, the observation group was additionally administered GDFMD orally. Each treatment course lasted eight days, for a total of four treatment courses. Efficacy evaluations were performed before treatment and after the second and fourth treatment courses, respectively. The clinical efficacy and safety of GDFMD in the treatment of WD-related liver fibrosis were assessed by comparing the changes in liver stiffness measurement (LSM), liver serological markers [alanine aminotransferase (ALT), aspartate aminotransferase (AST), type Ⅳ collagen (C-Ⅳ), laminin (LN), N-terminal propeptide of type Ⅲ procollagen (PⅢNP), and hyaluronic acid (HA)], fibrosis index based on 4 factors (FIB-4), AST to platelet ratio index (APRI), unified Wilson's disease rating scale part Ⅱ (UWDRS-Ⅱ), traditional Chinese medicine (TCM) syndrome score, 24-hour urinary copper, and safety indicators between the two groups before and after treatment. ResultsCompared with those before treatment, LSM levels decreased in both groups after two and four treatment courses (P<0.05). Compared with those after treatment, there was no statistically significant difference in the improvement of LSM levels in the observation group after two treatment courses, and the improvement of LSM levels in the observation group was more obvious after four treatment courses (P<0.05). Compared with those before treatment, the levels of HA, LN, PⅢNP, and C-Ⅳ decreased in both groups after two and four treatment courses (P<0.05). Compared with those after treatment, there was no statistically significant difference in the improvement of the C-Ⅳ levels in the observation group after two treatment courses, and the levels of HA, LN, and PⅢNP were more obvious (P<0.05). After four treatment courses in the observation group, the levels of HA, LN, PⅢNP, and C-Ⅳ were improved more significantly (P<0.05). Compared with those before treatment, ALT and AST levels decreased in both groups after two and four treatment courses (P<0.05). Compared with the control group after treatment, there was no statistically significant difference in the improvement of ALT and AST levels in the observation group after two treatment courses, and the improvement of ALT and AST levels in the observation group was more obvious after four treatment courses (P<0.05). Compared with those before treatment, APRI score and FIB-4 index level decreased in both groups after two and four treatment courses (P<0.05). Compared with those in control group after treatment, there was no statistically significant difference in the improvement of APRI score and FIB-4 index level in the observation group after two treatment courses, and the APRI score in the observation group was more obvious after four treatment courses (P<0.05), with no statistically significant improvement in the FIB-4 index difference. Compared with those before treatment, the levels of TCM syndrome scores decreased in both groups after two and four treatment courses (P<0.05). Compared with that of the control group after treatment, there was no statistically significant difference in the improvement of the level of TCM syndrome scores in the observation group after two treatment courses, and the improvement of the level of TCM syndrome scores in the observation group was more obvious after four treatment courses (P<0.05). Compared with those before treatment, the UWDRS-Ⅱ scores in both groups after two treatment courses were not improved obviously, and the UWDRS-Ⅱ scores in both groups decreased after four treatment courses (P<0.05). Compared with those of the control group after treatment, there was no statistically significant difference in the improvement of the UWDRS-Ⅱ scores in the observation group after two treatment courses, and the improvement of the UWDRS-Ⅱ scores in the observation group after four treatment courses was more obvious (P<0.05). Compared with those before treatment, the 24-h urine copper levels were significantly higher in both groups after two and four treatment courses (P<0.05). Compared with those in the control group after treatment, the 24-h urine copper levels in the observation group were significantly higher after two and four treatment courses (P<0.01). After two treatment courses, the 24-h urine copper level in the observation group showed a gradual decreasing trend, although it was higher than that before treatment. After four treatment courses, the control group had an improvement rate of 91.43%, an effective rate of 34.29%, and an apparent rate of 2.86%. The observation group had an improvement rate of 94.29%, an effective rate of 71.43%, and an apparent rate of 8.57%. The efficacy of the observation group was better than that of the control group (P<0.05). Conclusion① The efficacy of GDFMD combined with DMPS therapy in patients with WD-related liver fibrosis of liver-kidney deficiency and phlegm-blood stasis syndrome is significantly better than that of single DMPS therapy, and the advantages of the combined therapy are more obvious with the prolongation of the treatment cycle. ② GDFMD combined with the DMPS therapy program in the long-term application exhibits no obvious adverse reactions with good safety, which is worthy of clinical popularization and application.
4.Clinical Efficacy of Gandou Fumu Decoction in Treating Hepatolenticular Degeneration with Liver Fibrosis of Liver-kidney Deficiency and Phlegm-blood Stasis Syndrome
Pingping YANG ; Meixia WANG ; Changchang CAO ; Zhuang TAO ; Jiang DU ; Yun XU ; Wenming YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):182-191
ObjectiveThis paper aims to evaluate the intervention effect of Gandou Fumu Decoction (GDFMD) in treating hepatolenticular degeneration with liver fibrosis of liver-kidney deficiency and phlegm-blood stasis syndrome, thereby providing evidence-based medical evidence for the treatment of Wilson's disease (WD)-related liver fibrosis with traditional Chinese medicine through clinical efficacy analysis. MethodsA total of 70 patients with WD-related liver fibrosis of liver-kidney deficiency and phlegm-blood stasis syndrome meeting the inclusion criteria were enrolled from Anhui Provincial Hospital of TCM from October 1, 2023, to October 1, 2024. Participants were divided into a control group and an observation group, with 35 cases in each group. The control group received conventional copper chelation therapy with sodium dimercaptopropanesulfonate (DMPS). On this basis, the observation group was additionally administered GDFMD orally. Each treatment course lasted eight days, for a total of four treatment courses. Efficacy evaluations were performed before treatment and after the second and fourth treatment courses, respectively. The clinical efficacy and safety of GDFMD in the treatment of WD-related liver fibrosis were assessed by comparing the changes in liver stiffness measurement (LSM), liver serological markers [alanine aminotransferase (ALT), aspartate aminotransferase (AST), type Ⅳ collagen (C-Ⅳ), laminin (LN), N-terminal propeptide of type Ⅲ procollagen (PⅢNP), and hyaluronic acid (HA)], fibrosis index based on 4 factors (FIB-4), AST to platelet ratio index (APRI), unified Wilson's disease rating scale part Ⅱ (UWDRS-Ⅱ), traditional Chinese medicine (TCM) syndrome score, 24-hour urinary copper, and safety indicators between the two groups before and after treatment. ResultsCompared with those before treatment, LSM levels decreased in both groups after two and four treatment courses (P<0.05). Compared with those after treatment, there was no statistically significant difference in the improvement of LSM levels in the observation group after two treatment courses, and the improvement of LSM levels in the observation group was more obvious after four treatment courses (P<0.05). Compared with those before treatment, the levels of HA, LN, PⅢNP, and C-Ⅳ decreased in both groups after two and four treatment courses (P<0.05). Compared with those after treatment, there was no statistically significant difference in the improvement of the C-Ⅳ levels in the observation group after two treatment courses, and the levels of HA, LN, and PⅢNP were more obvious (P<0.05). After four treatment courses in the observation group, the levels of HA, LN, PⅢNP, and C-Ⅳ were improved more significantly (P<0.05). Compared with those before treatment, ALT and AST levels decreased in both groups after two and four treatment courses (P<0.05). Compared with the control group after treatment, there was no statistically significant difference in the improvement of ALT and AST levels in the observation group after two treatment courses, and the improvement of ALT and AST levels in the observation group was more obvious after four treatment courses (P<0.05). Compared with those before treatment, APRI score and FIB-4 index level decreased in both groups after two and four treatment courses (P<0.05). Compared with those in control group after treatment, there was no statistically significant difference in the improvement of APRI score and FIB-4 index level in the observation group after two treatment courses, and the APRI score in the observation group was more obvious after four treatment courses (P<0.05), with no statistically significant improvement in the FIB-4 index difference. Compared with those before treatment, the levels of TCM syndrome scores decreased in both groups after two and four treatment courses (P<0.05). Compared with that of the control group after treatment, there was no statistically significant difference in the improvement of the level of TCM syndrome scores in the observation group after two treatment courses, and the improvement of the level of TCM syndrome scores in the observation group was more obvious after four treatment courses (P<0.05). Compared with those before treatment, the UWDRS-Ⅱ scores in both groups after two treatment courses were not improved obviously, and the UWDRS-Ⅱ scores in both groups decreased after four treatment courses (P<0.05). Compared with those of the control group after treatment, there was no statistically significant difference in the improvement of the UWDRS-Ⅱ scores in the observation group after two treatment courses, and the improvement of the UWDRS-Ⅱ scores in the observation group after four treatment courses was more obvious (P<0.05). Compared with those before treatment, the 24-h urine copper levels were significantly higher in both groups after two and four treatment courses (P<0.05). Compared with those in the control group after treatment, the 24-h urine copper levels in the observation group were significantly higher after two and four treatment courses (P<0.01). After two treatment courses, the 24-h urine copper level in the observation group showed a gradual decreasing trend, although it was higher than that before treatment. After four treatment courses, the control group had an improvement rate of 91.43%, an effective rate of 34.29%, and an apparent rate of 2.86%. The observation group had an improvement rate of 94.29%, an effective rate of 71.43%, and an apparent rate of 8.57%. The efficacy of the observation group was better than that of the control group (P<0.05). Conclusion① The efficacy of GDFMD combined with DMPS therapy in patients with WD-related liver fibrosis of liver-kidney deficiency and phlegm-blood stasis syndrome is significantly better than that of single DMPS therapy, and the advantages of the combined therapy are more obvious with the prolongation of the treatment cycle. ② GDFMD combined with the DMPS therapy program in the long-term application exhibits no obvious adverse reactions with good safety, which is worthy of clinical popularization and application.
5.Traditional Chinese medicine syndrome and syndrome differentiation-based treatment of Wilson disease
Wenjie HAO ; Wenming YANG ; Ting CHENG ; Hailin JIANG ; Han WANG ; Meixia WANG
Journal of Clinical Hepatology 2026;42(3):522-528
Wilson disease (WD) is an autosomal recessive disorder of copper metabolism, and decoppering therapy and symptomatic treatment are the main Western medicine therapies for WD. This article systematically reviews the understanding of the etiology and pathogenesis of WD in traditional Chinese medicine (TCM) and points out that abnormal natural endowment is the core etiology and pathogenesis of WD, with internal accumulation of copper toxicity as the manifestation, liver/spleen/kidney dysfunction as the root cause, and intermingled “toxin, stasis, phlegm, and deficiency” as the key pathogenesis. Literature research and clinical observation are conducted to summarize the common TCM syndromes of WD, including stagnation of liver Qi, internal retention of damp-heat, phlegm-stasis-heat accumulation syndrome, liver-kidney Yin deficiency syndrome, spleen-kidney Yang deficiency, and syndrome of deficiency damage and phlegm stasis. This article proposes the corresponding therapies and representative prescriptions for each syndrome and discusses the advantages of treatment by stage and integrated traditional Chinese and Western medicine therapy. This article aims to provide a systematic reference for the syndrome differentiation-based treatment of WD in clinical practice of TCM, thereby giving full play to the advantages of TCM in the treatment of this disease.
6.Preliminary establishment of comprehensive risk assessment indicator system for occupational health based on Delphi method and analytic hierarchy process
Weiqi SUN ; Meixia LIU ; Tongning GAO ; Yuqi CHEN ; Yan YIN
Journal of Environmental and Occupational Medicine 2026;43(6):754-761
Background Occupational health risk assessment is the foundation of safeguarding workers' occupational health. However, traditional assessment methods often fail to adequately account for health effects and enterprise-specific characteristics, limiting their comprehensive evaluation capabilities. Additionally, these methods suffer from inconsistent standards, narrow application scopes, insufficient hazard sub-categorization, and indicator systems that poorly support informatization and digital transformation. To address these gaps, this study proposes methodological innovations focusing on the assessment indicator system. Objective To construct a systematic, generalizable, and practical occupational health comprehensive risk assessment indicator system by leveraging occupational health practices and surveillance big data from Shanghai. Methods Initial system indicators were extrated through literature review. Subsequently, expert interviews and group discussions, combined with occupational health surveillance big data, were used to refine these indicators, forming a candidate pool covering two major dimensions: risk factor characteristics and enterprise characteristics. The Delphi method was applied to evaluate and screen the indicators, while the analytic hierarchy process (AHP) was used to construct a hierarchical model and determine indicator weights. Results A total of 76 articles were reviewed, identifying 25 candidate indicators. After two rounds of Delphi expert consultation, 8 primary indicators (e.g. hazard characteristics, exposure characteristics, health effects, industry characteristics, management characteristics, protective features, adverse events, and uncertainty) and 17 secondary indicators (hazard factor characteristics, exposure levels, number of people exposed, number of factor exposure, special physical exam indicators, suspected occupational diseases, occupational diseases, industry categories, organizational management, hazard reporting, occupational health training, hazard factor testing, occupational health exams, protective facilities, personal protective equipment, administrative penalties, and data quality) were finalized. The expert response rate was 100%, and the expert authority coefficient was 0.86. The Kendall's W coefficients for importance and accessibility scores across the two consultation rounds were 0.41, 0.40 and 0.41, 0.50. A hierarchical model targeting the comprehensive occupational health risk index was constructed, comprising two major elements and three levels of indicators. AHP was applied to calculate the weights, with all judgment matrices demonstrating satisfactory consistency (consistency ratio < 0.1). Conclusion By integrating risk factor profiles with enterprise-specific characteristics via the Delphi-AHP approach, a robust occupational health comprehensive risk assessment indicator system is established. This system provides a practical framework for digitized occupational health management, though further empirical validation in real-world settings is warranted.
7.Predicting Hepatic Fibrosis Risk in Wilson's Disease: Development and Validation of Prognostic Model Integrating TCM Syndromes and Modern Biomarkers
Jiafeng ZHOU ; Meixia WANG ; Zhuang TAO ; Shuai KANG ; Gang WANG ; Rui WANG ; Wenming YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(18):197-207
ObjectiveTo analyze the influencing factors of hepatic fibrosis in patients with Wilson's disease (WD) and to develop and validate a predictive model for hepatic fibrosis in WD that integrates traditional Chinese medicine (TCM) syndromes and modern biomarkers, thus providing a basis for early identification of high-risk populations and medical intervention. MethodsThe clinical data of 220 WD patients diagnosed in the Department of Encephalopathy, The First Affiliated Hospital of Anhui University of Chinese Medicine between January 2010 and December 2024 were retrospectively collected. Through a random number table, patients were assigned into a training set (154 patients) and a validation set (66 patients) in a 7∶3 ratio. Univariate COX regression, LASSO regression analysis, and multivariate COX regression analysis were performed to identify independent influencing factors for hepatic fibrosis in WD patients, and a clinical prediction model was established based on these factors. The discrimination, calibration, and clinical utility of the model were evaluated based on the area under the receiver operating characteristic curve (AUC), calibration curves, and decision curve analysis (DCA), respectively. ResultsA total of 220 patients were included. The syndrome of combined phlegm and stasis, triglycerides (TG), laminin (LN), and male gender were identified as independent risk factors for hepatic fibrosis in WD patients, while TCM intervention and high-density lipoprotein cholesterol (HDL-C) were protective factors. The C-index indicated excellent discriminative ability (training set: 3-year AUC=0.908, 5-year AUC=0.869, 7-year AUC=0.829; validation set: 3-year AUC= 0.898, 5-year AUC=0.780, 7-year AUC=0.743). Calibration curves showed good consistency. DCA demonstrated the clinical net benefit probabilities across different risk thresholds at various time points. ConclusionThe predictive model established in this study has high accuracy and can be conveniently used for the early identification and risk prediction of hepatic fibrosis in patients with WD.
8.Gantou Fumu Decoction Inhibits Hepatic Stellate Cell Activation by Regulating Nrf2/ARE Signaling Pathway
Wangsheng HE ; Yanping ZHU ; Shizhen LONG ; Mengmeng LI ; Wenming YANG ; Han WANG ; Meixia WANG ; Ting DONG ; Shupei JIA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(23):105-115
ObjectiveTo explore the mechanism through which Gantou Fumu decoction (GDFMT)-containing serum inhibits the activation of hepatic stellate cells. MethodsSerum pharmacology techniques were employed to prepare GDFMT-containing serum samples. Rat hepatic stellate cells (HSCT6) were randomly allocated into three groups: Control, penicillamine (90 mg·kg-1), and GDFMT (6.8 g·kg-1). Cell viability was assessed by the cell counting kti-8 (CCK-8) assay, intracellular ROS levels were determined by flow cytometry using the DCFH‑DA fluorescent probe and while extracellular collagen Ⅰ (ColⅠ) levels were quantified via enzyme linked immunosorbent assay (ELISA). Intracellular α-smooth muscle actin (α-SMA) expression was visualized through immunofluorescence staining. Real-time polymerase chain reaction(Real-time PCR) and Western blot were performed to evaluate alterations in the expression of nuclear factor-erythroid 2-related factor 2 (Nrf2), glutamate cysteine ligase, catalytic (GCLC), glutamate-cysteine ligase, modifier subunit (GCLM), Kelch-like ECH-associated protein 1 (Keap1), NAD(P)H:Quinone oxidoreductase 1 (NQO1), and heme oxygenase-1 (HO-1). Genetic knockdown of Nrf2 was achieved through small interfering (siRNA) transfection to validate pathway specificity. ResultsCCK-8 data indicated that under the selected experimental conditions, culturing HSCT6 cells with 10% (volume fraction) GDFMT-containing serum for 48 h demonstrated equivalent effect in inhibiting HSCT6 cell proliferation compared with culturing HSCT6 cells with 10% (volume fraction) penicillamine-containing serum for 36 h, with no statistically significant difference between the two groups. After treatment of HSCT6 cells with siRNA-mediated Nrf2 gene transfection technique, the inhibitory effect of GDFMT on the activation process of HSCT6 cells was significantly weakened. After HSCT6 cells were cultured with GDFMT-containing serum for 48 h, the flow cytometry with a fluorescence probe showed that the content of reactive oxygen species (ROS) in the cells was significantly reduced. After the expression level of the Nrf2 gene in HSCT6 cells was knocked down by siRNA transfection, the content of ROS increased (P<0.01), indicating that GDFMT significantly reduced the ROS level in activated HSCT6 cells, and this effect strictly depended on the integrity of the Nrf2 pathway. Quantitative analysis by ELISA showed that the synthesis of ColⅠ in the HSCT6 cell culture system treated with GDFMT-containing serum for 48 h was lower than that in the control group (P<0.01). After the Nrf2 gene level of HSCT6 cells was knocked down by siRNA transfection, the ColⅠ concentration in the culture supernatant increased (P<0.01). The results of α-SMA detection by IF method showed that compared with the normal group, GDFMT-containing serum reduced the expression of α-SMA (P<0.01) and the protein level of α-SMA in HSCT6 cells was increased (P<0.01) after siRNA-mediated Nrf2 silencing, suggesting that Nrf2 signaling pathway may be involved in regulating the phenotypic transformation process of the cells. Real-time PCR and Western blot results showed that GDFMT-containing serum reduced the protein level of α-SMA (P<0.01) and enhanced the gene and protein levels of Nrf2 in the nucleus and GCLC, HO-1, Keap1, GCLM, and NQO1 in the cytoplasm of HSCT6 cells (P<0.05, P<0.01). After the Nrf2 gene level of HSCT6 cells was knocked down by siRNA transfection, the protein level of α-SMA was increased (P<0.01), and the levels of GCLC, GCLM, Keap1, NQO1, and HO-1 were decreased (P<0.05, P<0.01). These results indicated that GDFMT-containing serum successfully triggered the Nrf2 signaling pathway of HSCT6 cells. ConclusionThe anti-hepatic fibrosis effect of 10%GDFMT was equivalent to that of penicillamine. In addition, GDFMT can inhibit HSC activation and reduce oxidative stress damage by activating the Nrf2/antioxidant response element (ARE) pathway, as manifested by the up-regulated expression of Nrf2 and downstream antioxidant genes (GCLC, GCLM, HO-1, Keap1, and NQO1) and reduced secretion of ROS, α-SMA, and ColⅠ.
9.Study on the relationship between serum SAA,IL-10,IL-21 levels and the first acute exacerbation in COPD patients with pneumothorax
Yumei CHEN ; Meixia ZHANG ; Maoliang TIAN
International Journal of Laboratory Medicine 2025;46(15):1825-1830
Objective To investigate the relationship between serum amyloid A(SAA),interleukin-10(IL-10),interleukin-21(IL-21)levels and the first acute exacerbation in patients with chronic obstructive pulmo-nary disease(COPD)with pneumothorax.Methods A total of 102 patients with stable COPD complicated with pneumothorax admitted to the hospital from January 2020 to January 2023 were selected.They were treated with closed thoracic drainage,anti-inflammatory,antiasthmatic,expectorant,and high-flow nasal hu-midification oxygen therapy(HFNC).The patients were divided into two groups according to whether they had the first acute exacerbation within 1 year after treatment,the first acute exacerbation group(32 patients had the first acute exacerbation within 1 year of follow-up)and the non-first acute exacerbation group(70 pa-tients had no first acute exacerbation within 1 year of follow-up).Serum SAA,IL-10 and IL-21 levels and blood gas indexes were compared between the two groups after treatment.Multivariate Logistic regression a-nalysis was used to analyze the influencing factors of the first acute exacerbation after HFNC treatment.The receiver operating characteristic(ROC)curve was used to analyze the predictive value of serum SAA,IL-10 and IL-21 levels after treatment for the first acute exacerbation of COPD patients with pneumothorax.Results There were significant differences in hypoproteinemia,smoking,underlying diseases,forced vital ca-pacity(FVC),forced expiratory volume in one second(FEV1)as a percentage of predicted value(FEV1%),FEV1 to FVC ratio(FEV1/FVC),arterial partial pressure of oxygen(PaO2),arterial partial pressure of car-bon dioxide(PaCO2),SAA,IL-10,IL-21 between the two groups(P<0.05).Multivariate Logistic regression analysis showed that hypoproteinemia,smoking,underlying diseases,PaCO2,SAA,IL-10 and IL-21 were the risk factors for the first acute exacerbation after HFNC treatment(P<0.05).FVC,FEV1%,FEV1/FVC and PaO2 were protective factors for the first acute exacerbation after treatment(P<0.05).ROC curve results showed that the area under the curve(AUC)of SAA,IL-10 and IL-21 levels after treatment to predict the first acute exacerbation of COPD patients with pneumothorax was 0.755,0.726 and 0.674,respectively.When the cut-off values of SAA,IL-10 and IL-21 were 171.06 g/L,26.46 pg/mL and 244.79 pg/mL,the sen-sitivity was 76.51%,60.84%and 56.90%,and the specificity was 66.73%,74.49%and 74.52%,respective-ly.The AUC of combined prediction was 0.860,the sensitivity was 80.44%,and the specificity was 76.51%.Conclusion The serum SAA,IL-10 and IL-21 levels in COPD patients with pneumothorax have certain pre-dictive value for the first acute exacerbation of COPD patients with pneumothorax.
10.Clinical value of quality control circle activities in preventing deep vein thrombosis in military hospitals
Xile WEI ; Xuehui HU ; Hange SONG ; Meixia ZHANG ; Guang CHENG ; Heng LUO ; Desheng WANG
Journal of Navy Medicine 2025;46(2):150-155
Objective To explore the clinical effect of quality control circle(QCC)activities on the prevention of postoperative deep vein thrombosis(DVT)in inpatients in military hospitals.Methods A total of 318 patients who were diagnosed and treated in The First Affiliated Hospital of Air Force Medical University from January to December 2021 and 40 medical staff were enrolled in this study.Routine care was performed in 158 patients from January to June 2021,and QCC care was implemented in 160 patients from July to December 2021.The awareness of DVT prevention in medical staff and patients(or their famiy members)before and after QCC activities,lower limb DVT preventive measures taken by medical staff,and the occurrence of DVT were compared.Results The scores of the cause,clinical manifestations,nursing measures and preventive measures of DVT after QCC activities were significantly higher than those before QCC activities in both medical staff and patients(or their families)(P<0.05).The overall implementation rate of preventing lower limb DVT after QCC activities was significantly higher than that before QCC activities(94.14%vs.46.20%,P<0.05).The incidence of DVT after QCC activities was significantly lower than that before QCC activities(0.62%vs.5.06%,P<0.05).Conclusion Implementing QCC activities can improve the cognitive ability of military medical staff and patients(or family members)in DVT prevention,increase the implementation rate of DVT prevention measures,and reduce the incidence of DVT.

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