ObjectiveTo construct a competing endogenous RNA （ceRNA） network involving long non-coding RNA NOTCH1 （lncRNA NOTCH1） and investigate whether Xianglian Huazhuo prescription （XLHZ） ameliorates gastric mucosal injury via regulating ferroptosis through the lncRNA NOTCH1/microRNA-138-5p （miR-138-5p）/silent information regulator 1 （SIRT1） axis. MethodsLncRNA sequencing and bioinformatics tools were used to construct the ceRNA network involving lncRNA NOTCH1. In the animal experiments， a chronic atrophic gastritis （CAG） model was induced by N-methyl-N'-nitro-N-nitrosoguanidine （MNNG） in mice. Mice were randomized into normal， model， Morodan （2.0 g·kg^-1）， and high-， medium-， and low-dose （7.6， 3.8， 1.9 g·kg^-1， respectively） XLHZ groups （n=10）. Transmission electron microscopy was adopted to assess gastric pathology. Real-time PCR was employed to measure the mRNA levels of lncRNA NOTCH1， miR-138-5p， SIRT1， nuclear factor erythroid 2-related factor 2 （Nrf2）， and glutathione peroxidase 4 （GPX4）. Western blot was adopted to quantify the protein levels of SIRT1， Nrf2， and GPX4 in the gastric tissue. In the cell experiment， dual-luciferase assay was employed to verify the targeting among lncRNA NOTCH1， miR-138-5p， and SIRT1. MNNG was used to treat GES-1 cells for disease modeling. The effects of NOTCH1 mimics on the expression of lncRNA NOTCH1， miR-138-5p， and SIRT1， as well as the effects of miR-138-5p inhibitor on the expression of miR-138-5p and SIRT1， were observed. The rescue experiment with XLHZ was designed with five groups： GES-1， GES-1+MNNG （Model）， Model+XLHZ， Model+NOTCH1 inhibitor+XLHZ， and Model+NOTCH1 inhibitor+miR-138-5p mimics+XLHZ. Real-time PCR and immunofluorescence assay were employed to measure the mRNA levels of lncRNA NOTCH1， miR-138-5p， SIRT1， Nrf2， and GPX4 and the Fe²⁺ content， respectively， in GES-1 cells. ResultsThe modeled mice showed mitochondrial damage with shrinkage and cristae loss， downregulated mRNA/protein levels of lncRNA NOTCH1， SIRT1， Nrf2， GPX4， and upregulated expression of miR-138-5p （P<0.01）. Compared with the model group， drug treatments reduced mitochondrial structural damage and the membrane density decreased， partially recovered the crista structure， downregulated the expression of miR-138-5p， and upregulated the mRNA and protein levels of lncRNA NOTCH1， SIRT1， Nrf2， and GPX4 （P<0.05， P<0.01）. In the cell experiments， the dual-luciferase assays confirmed lncRNA NOTCH1 bound to miR-138-5p and miR-138-5p bound to SIRT1， both suppressing luciferase activity （P<0.05， P<0.01）. Compared with the blank control group， the model group showed decreased expression of lncRNA NOTCH1， SIRT1， Nrf2， GPX4， increased expression of miR-138-5p， and enhanced Fe²⁺ fluorescence （P<0.01）. NOTCH1 mimics or miR-138-5p inhibitor reversed these changes （P<0.05， P<0.01）. Rescue experiments showed that NOTCH1 inhibitor reversed XLHZ-induced miR-138-5p decrease and SIRT1 increase （P<0.01）， while miR-138-5p overexpression suppressed SIRT1 expression （P<0.05， P<0.01）. The results confirmed that lncRNA NOTCH1 negatively regulated miR-138-5p， which in turn negatively regulated SIRT1. ConclusionXLHZ effectively alleviates gastric mucosal injury and curbs CAG progression by inhibiting ferroptosis through modulation of the lncRNA NOTCH1/miR-138-5p/SIRT1 axis.

ObjectiveTo explore the mechanism of Xianglian Huazhuo prescription （XLHZ） in ameliorating chronic atrophic gastritis （CAG） by regulating lysosomal iron metabolism. MethodsIn the animal experiments， a mouse model of CAG was established through a combined modeling approach， including free drinking of N-methyl-N′-nitro-N-nitrosoguanidine （MNNG， 100 mg·L^-1）， gavage of ranitidine （8 g·L^-1）， and irregular feeding. Mice were randomly assigned into the normal， model， Morodan （2.0 g·kg^-1）， and high-， medium-， and low-dose （7.2， 3.8， 1.9 g·kg^-1， respectively） XLHZ groups. In the cell experiments， GES-1 cells were treated with MNNG to construct a cell model of CAG， and cells were allocated into the GES-1， GES-1+MNNG （Model）， and Model+XLHZ groups. The general physical conditions of mice in each group were observed. Hematoxylin-eosin （HE） staining was used to detect the pathological morphology of the gastric mucosa. Transmission electron microscopy was employed to observe the ultrastructure of gastric mucosal cells. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the serum levels of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and interleukin-10 （IL-10） in the serum. Western blot was used to determine the protein levels of lysosome-associated membrane protein 1 （LAMP1）， nuclear receptor coactivator 4 （NCOA4）， ferritin heavy chain 1 （FTH1）， six-transmembrane epithelial antigen of prostate 3 （Steap3）， and transient receptor potential mucolipin 1 （TRPML1） in the gastric tissue. Immunohistochemistry was employed to detect the positive expression of glutathione peroxidase 4 （GPX4） and 8-hydroxy-2′-deoxyguanosine （8-OHdG） in the gastric tissue. Fluorescent probe method was used to detect the levels of Fe²⁺ and reactive oxygen species （ROS）. Immunofluorescence assay was applied to observe the co-localization of FTH1 and LAMP1. ResultsThe animal experiments showed that compared with the normal group， the model group exhibited poor mental status and hair condition， accompanied by decreased locomotor activity， food and water intake， and even reduced body weight. HE staining results showed that the gastric mucosal epithelium of model mice presented poor continuity， significantly decreased mucosal thickness， irregular gastric fovea morphology with widened spacing， dilated glandular cavities， reduced and disarranged glands with obvious atrophy， and extensive inflammatory cell infiltration. Transmission electron microscopy revealed that the mitochondria were markedly shrunken with condensed and deeply stained matrix， drastically reduced and irregularly arranged cristae， and even vacuolization， and the outer mitochondrial membrane shrank and ruptured. In addition， the model group showed elevated serum levels of IL-6 and IL-1β （P<0.01）， declined serum level of IL-10 （P<0.01）， upregulated protein levels of LAMP1， NCOA4， Steap3， and TRPML1 （P<0.01）， downregulated protein level of FTH1 （P<0.01） in the gastric tissue， reduced positive expression of GPX4 （P<0.01）， and increased positive expression of 8-OHdG （P<0.01）. Compared with the model group， all the treatment groups showed improved mental state and hair appearance， increased locomotor activity， food and water intake， as well as gradual body weight gain in mice. HE staining demonstrated that the gastric mucosal epithelium of mice in each treatment group was continuous and intact， with normalized gastric fovea morphology， narrowed glandular cavities， increased and neatly arranged glands， and alleviated inflammatory cell infiltration. Transmission electron microscopy results showed that the mitochondria in mice from each treatment group possessed intact structures with recovered normal volume， increased and regularly arranged cristae， and homogeneous density of the mitochondrial membrane. The mice treated with high and medium doses of XLHZ exhibited declined serum levels of IL-6 and IL-1β （P<0.01） and elevated serum level of IL-10 （P<0.01）. Low-dose XLHZ reduced serum IL-1β level in mice （P<0.05）. The expression levels of LAMP1， NCOA4， Steap3 and TRPML1 protein in gastric tissue of mice in XLHZ high- and medium-dose groups were decreased （P<0.05， P<0.01）， and the expression level of FTH1 protein was increased （P<0.05， P<0.01）. The positive expression of GPX4 in the gastric tissue was increased in the high- and medium-dose XLHZ groups （P<0.05， P<0.01）， and that of 8-OHdG was decreased in all treatment groups （P<0.01）. The cell experiments showed that compared with the GES-1 group， the fluorescence intensities of Fe²⁺ and ROS， as well as the co-localization of FTH1 and LAMP1， were significantly increased in the GES-1 + MNNG group （P<0.01）. Compared with the GES-1 + MNNG group， the fluorescence intensities of Fe²⁺ and ROS were markedly decreased in the Model + XLHZ group （P<0.01）， while the co-localization of FTH1 and LAMP1 was reduced （P<0.05）. ConclusionXLHZ can effectively regulate lysosomal iron metabolism and ameliorate gastric mucosal and cellular injury induced by CAG.

ObjectiveTo construct a competing endogenous RNA （ceRNA） network involving long non-coding RNA NOTCH1 （lncRNA NOTCH1） and investigate whether Xianglian Huazhuo prescription （XLHZ） ameliorates gastric mucosal injury via regulating ferroptosis through the lncRNA NOTCH1/microRNA-138-5p （miR-138-5p）/silent information regulator 1 （SIRT1） axis. MethodsLncRNA sequencing and bioinformatics tools were used to construct the ceRNA network involving lncRNA NOTCH1. In the animal experiments， a chronic atrophic gastritis （CAG） model was induced by N-methyl-N'-nitro-N-nitrosoguanidine （MNNG） in mice. Mice were randomized into normal， model， Morodan （2.0 g·kg^-1）， and high-， medium-， and low-dose （7.6， 3.8， 1.9 g·kg^-1， respectively） XLHZ groups （n=10）. Transmission electron microscopy was adopted to assess gastric pathology. Real-time PCR was employed to measure the mRNA levels of lncRNA NOTCH1， miR-138-5p， SIRT1， nuclear factor erythroid 2-related factor 2 （Nrf2）， and glutathione peroxidase 4 （GPX4）. Western blot was adopted to quantify the protein levels of SIRT1， Nrf2， and GPX4 in the gastric tissue. In the cell experiment， dual-luciferase assay was employed to verify the targeting among lncRNA NOTCH1， miR-138-5p， and SIRT1. MNNG was used to treat GES-1 cells for disease modeling. The effects of NOTCH1 mimics on the expression of lncRNA NOTCH1， miR-138-5p， and SIRT1， as well as the effects of miR-138-5p inhibitor on the expression of miR-138-5p and SIRT1， were observed. The rescue experiment with XLHZ was designed with five groups： GES-1， GES-1+MNNG （Model）， Model+XLHZ， Model+NOTCH1 inhibitor+XLHZ， and Model+NOTCH1 inhibitor+miR-138-5p mimics+XLHZ. Real-time PCR and immunofluorescence assay were employed to measure the mRNA levels of lncRNA NOTCH1， miR-138-5p， SIRT1， Nrf2， and GPX4 and the Fe²⁺ content， respectively， in GES-1 cells. ResultsThe modeled mice showed mitochondrial damage with shrinkage and cristae loss， downregulated mRNA/protein levels of lncRNA NOTCH1， SIRT1， Nrf2， GPX4， and upregulated expression of miR-138-5p （P<0.01）. Compared with the model group， drug treatments reduced mitochondrial structural damage and the membrane density decreased， partially recovered the crista structure， downregulated the expression of miR-138-5p， and upregulated the mRNA and protein levels of lncRNA NOTCH1， SIRT1， Nrf2， and GPX4 （P<0.05， P<0.01）. In the cell experiments， the dual-luciferase assays confirmed lncRNA NOTCH1 bound to miR-138-5p and miR-138-5p bound to SIRT1， both suppressing luciferase activity （P<0.05， P<0.01）. Compared with the blank control group， the model group showed decreased expression of lncRNA NOTCH1， SIRT1， Nrf2， GPX4， increased expression of miR-138-5p， and enhanced Fe²⁺ fluorescence （P<0.01）. NOTCH1 mimics or miR-138-5p inhibitor reversed these changes （P<0.05， P<0.01）. Rescue experiments showed that NOTCH1 inhibitor reversed XLHZ-induced miR-138-5p decrease and SIRT1 increase （P<0.01）， while miR-138-5p overexpression suppressed SIRT1 expression （P<0.05， P<0.01）. The results confirmed that lncRNA NOTCH1 negatively regulated miR-138-5p， which in turn negatively regulated SIRT1. ConclusionXLHZ effectively alleviates gastric mucosal injury and curbs CAG progression by inhibiting ferroptosis through modulation of the lncRNA NOTCH1/miR-138-5p/SIRT1 axis.

ObjectiveTo explore the mechanism of Xianglian Huazhuo prescription （XLHZ） in ameliorating chronic atrophic gastritis （CAG） by regulating lysosomal iron metabolism. MethodsIn the animal experiments， a mouse model of CAG was established through a combined modeling approach， including free drinking of N-methyl-N′-nitro-N-nitrosoguanidine （MNNG， 100 mg·L^-1）， gavage of ranitidine （8 g·L^-1）， and irregular feeding. Mice were randomly assigned into the normal， model， Morodan （2.0 g·kg^-1）， and high-， medium-， and low-dose （7.2， 3.8， 1.9 g·kg^-1， respectively） XLHZ groups. In the cell experiments， GES-1 cells were treated with MNNG to construct a cell model of CAG， and cells were allocated into the GES-1， GES-1+MNNG （Model）， and Model+XLHZ groups. The general physical conditions of mice in each group were observed. Hematoxylin-eosin （HE） staining was used to detect the pathological morphology of the gastric mucosa. Transmission electron microscopy was employed to observe the ultrastructure of gastric mucosal cells. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the serum levels of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and interleukin-10 （IL-10） in the serum. Western blot was used to determine the protein levels of lysosome-associated membrane protein 1 （LAMP1）， nuclear receptor coactivator 4 （NCOA4）， ferritin heavy chain 1 （FTH1）， six-transmembrane epithelial antigen of prostate 3 （Steap3）， and transient receptor potential mucolipin 1 （TRPML1） in the gastric tissue. Immunohistochemistry was employed to detect the positive expression of glutathione peroxidase 4 （GPX4） and 8-hydroxy-2′-deoxyguanosine （8-OHdG） in the gastric tissue. Fluorescent probe method was used to detect the levels of Fe²⁺ and reactive oxygen species （ROS）. Immunofluorescence assay was applied to observe the co-localization of FTH1 and LAMP1. ResultsThe animal experiments showed that compared with the normal group， the model group exhibited poor mental status and hair condition， accompanied by decreased locomotor activity， food and water intake， and even reduced body weight. HE staining results showed that the gastric mucosal epithelium of model mice presented poor continuity， significantly decreased mucosal thickness， irregular gastric fovea morphology with widened spacing， dilated glandular cavities， reduced and disarranged glands with obvious atrophy， and extensive inflammatory cell infiltration. Transmission electron microscopy revealed that the mitochondria were markedly shrunken with condensed and deeply stained matrix， drastically reduced and irregularly arranged cristae， and even vacuolization， and the outer mitochondrial membrane shrank and ruptured. In addition， the model group showed elevated serum levels of IL-6 and IL-1β （P<0.01）， declined serum level of IL-10 （P<0.01）， upregulated protein levels of LAMP1， NCOA4， Steap3， and TRPML1 （P<0.01）， downregulated protein level of FTH1 （P<0.01） in the gastric tissue， reduced positive expression of GPX4 （P<0.01）， and increased positive expression of 8-OHdG （P<0.01）. Compared with the model group， all the treatment groups showed improved mental state and hair appearance， increased locomotor activity， food and water intake， as well as gradual body weight gain in mice. HE staining demonstrated that the gastric mucosal epithelium of mice in each treatment group was continuous and intact， with normalized gastric fovea morphology， narrowed glandular cavities， increased and neatly arranged glands， and alleviated inflammatory cell infiltration. Transmission electron microscopy results showed that the mitochondria in mice from each treatment group possessed intact structures with recovered normal volume， increased and regularly arranged cristae， and homogeneous density of the mitochondrial membrane. The mice treated with high and medium doses of XLHZ exhibited declined serum levels of IL-6 and IL-1β （P<0.01） and elevated serum level of IL-10 （P<0.01）. Low-dose XLHZ reduced serum IL-1β level in mice （P<0.05）. The expression levels of LAMP1， NCOA4， Steap3 and TRPML1 protein in gastric tissue of mice in XLHZ high- and medium-dose groups were decreased （P<0.05， P<0.01）， and the expression level of FTH1 protein was increased （P<0.05， P<0.01）. The positive expression of GPX4 in the gastric tissue was increased in the high- and medium-dose XLHZ groups （P<0.05， P<0.01）， and that of 8-OHdG was decreased in all treatment groups （P<0.01）. The cell experiments showed that compared with the GES-1 group， the fluorescence intensities of Fe²⁺ and ROS， as well as the co-localization of FTH1 and LAMP1， were significantly increased in the GES-1 + MNNG group （P<0.01）. Compared with the GES-1 + MNNG group， the fluorescence intensities of Fe²⁺ and ROS were markedly decreased in the Model + XLHZ group （P<0.01）， while the co-localization of FTH1 and LAMP1 was reduced （P<0.05）. ConclusionXLHZ can effectively regulate lysosomal iron metabolism and ameliorate gastric mucosal and cellular injury induced by CAG.

Background/Aims: Metabolic dysfunction-associated steatohepatitis (MASH) is a significant risk factor for gallstone formation, but mechanisms underlying MASH-related gallstone formation remain unclear. Golgi membrane protein 1 (GOLM1) participates in hepatic cholesterol metabolism and is upregulated in MASH. Here, we aimed to explore the role of GOLM1 in MASH-related gallstone formation. Methods: The UK Biobank cohort was used for etiological analysis. GOLM1 knockout (GOLM1-/-) and wild-type (WT) mice were fed with a high-fat diet (HFD). Livers were excised for histology and immunohistochemistry analysis. Gallbladders were collected to calculate incidence of cholesterol gallstones (CGSs). Biles were collected for biliary lipid analysis. HepG2 cells were used to explore underlying mechanisms. Human liver samples were used for clinical validation. Results: MASH patients had a greater risk of cholelithiasis. All HFD-fed mice developed MASH, and the incidence of gallstones was 16.7% and 75.0% in GOLM1-/- and WT mice, respectively. GOLM1-/- decreased biliary cholesterol concentration and output. In vivo and in vitro assays confirmed that GOLM1 facilitated cholesterol efflux through upregulating ATP binding cassette transporter subfamily G member 5 (ABCG5). Mechanistically, GOLM1 translocated into nucleus to promote osteopontin (OPN) transcription, thus stimulating ABCG5-mediated cholesterol efflux. Moreover, GOLM1 was upregulated by interleukin-1β (IL-1β) in a dose-dependent manner. Finally, we confirmed that IL-1β, GOLM1, OPN, and ABCG5 were enhanced in livers of MASH patients with CGSs. Conclusions In MASH livers, upregulation of GOLM1 by IL-1β increases ABCG5-mediated cholesterol efflux in an OPN-dependent manner, promoting CGS formation. GOLM1 has the potential to be a molecular hub interconnecting MASH and CGSs.

BACKGROUND:Eucommia ulmoides has a certain osteogenic effect,which can promote the proliferation and differentiation of osteoblasts.However,it is unclear whether Eucommia ulmoides has effects on alveolar bone formation and Wnt/β-Catenin signaling pathway. OBJECTIVE:To investigate the mechanism by which Eucommia ulmoides promotes alveolar bone formation in ovariectomized rats based on the Wnt/β-Catenin signaling pathway. METHODS:Sixty female Sprague-Dawley rats were selected and randomly divided into five groups:blank control group,sham-operation group,model group,low-dose group Eucommia ulmoides group,and high-dose Eucommia ulmoides group,with twelve rats in each group.Osteoporosis animal models were constructed by bilateral oophorectomy in the model group and the low-dose and high-dose Eucommia ulmoides groups.The sham-operation group underwent the same method to remove adipose tissue of equal mass around the bilateral ovaries.Three months after surgery,the low-and high-dose Eucommia ulmoides groups were given 2.1 g/kg/d and 4.2 g/kg/d Eucommia ulmoides by gavage,respectively.The sham-operation group and model group were given the same amount of physiological saline by gavage.After 12 weeks of drug intervention,the changes in alveolar bone mass of rats in each group were observed through Micro-CT;hematoxylin-eosin staining was used to observe the pathological structural changes of alveolar bone in rats;enzyme linked immunosorbent assay was used to detect the expression levels of alkaline phosphatase and osteocalcin in the serum of rats;western blot was used to detect the expression levels of β-Catenin and Frizzled9 receptor proteins in the alveolar bone of rats;and real-time fluorescence quantitative PCR was used to detect the expression of osteocalcin,Runt-related transcription factor 2(Runx2),alkaline phosphatase,β-catenin,and frizzled9 mRNAs in alveolar bone tissues of rats. RESULTS AND CONCLUSION:Compared with the blank control group,bone volume fraction,trabecular number,trabecular thickness,and bone mineral density were reduced in the model group(P＜0.05),and trabecular separation was elevated(P＜0.05).Pathological observation showed that the arrangement of trabeculae was disordered and irregular,the trabeculae were thinned or broken,and the marrow cavity was enlarged in the model group,with a significant reduction in bone volume;the level of alkaline phosphatase in the serum was increased(P＜0.05),and the level of osteocalcin was decreased(P＜0.05);mRNA expression of alkaline phosphatase,osteocalcin,Runx2,β-catenin,and frizzled9 were decreased(P＜0.05);protein expression of β-Catenin and Frizzled9 was decreased(P＜0.05).Compared with the model group,the low-and high-dose Eucommia ulmoides groups showed an increase in bone volume fraction,trabecular number,trabecular thickness,and bone mineral density(P＜0.05)and a decrease in trabecular separation(P＜0.05).In the low-and high-dose Eucommia ulmoides groups,bone trabeculae were slightly aligned and thickened,with a significant increase in bone mass.Compared with the model group,the serum level of alkaline phosphatase was reduced(P＜0.05)and the serum level of osteocalcin was elevated(P＜0.05)in the low-and high-dose Eucommia ulmoides groups.Compared with the model group,the mRNA expression of alkaline phosphatase,osteocalcin,Runx2,β-catenin,and frizzled9 were increased in the low-and high-dose Eucommia ulmoides groups(P＜0.05).Compared with the model group,the protein expression of Frizzled9 was increased in the low-dose Eucommia ulmoides group(P＜0.05),while the protein expression of β-Catenin and Frizzled9 was increased in the high-dose Eucommia ulmoides group(P＜0.05).Compared with the low-dose Eucommia ulmoides group,the high-dose Eucommia ulmoides group had a more significant improvement in the above indexes.To conclude,Eucommia ulmoides can effectively promote the alveolar bone formation,and its mechanism of action might be related to the activation of the Wnt/β-catenin signaling pathway.

ObjectiveChinese patent medicines for cardiovascular and cerebrovascular diseases are diverse and complex in their efficacy. The traditional classification method based on efficacy categories has certain limitations and cannot meet the clinical needs for individualized drug selection and variety comparison. This article， based on the formulation compatibility analysis technology of "Tangye Jingfa Tu"， clarifies the composition and efficacy characteristics of common Chinese patent medicines used for cardiovascular and cerebrovascular diseases， providing support for the precise selection of these medicines. MethodsFifty-six representative Chinese patent medicines， covering all the efficacy subcategories of "stasis-resolving agents" in the National Basic Medical Insurance， Work Injury Insurance， and Maternity Insurance Drug Catalogue （2023） （more than 50% of the total）， were selected for the study. Within the knowledge system of "Tangye Jingfa Tu"， the compatibility structure of herbal flavors and the proportion structure of herbal quantities for each Chinese patent medicine were determined. The correlation between these structures and the efficacy categories was analyzed to identify the similarities and differences among the selected Chinese patent medicines. Additionally， the efficacy was reclassified and compared according to the theoretical framework of tonifying and purging methods of five Zang organs in the "Tangye Jingfa Tu". ResultsThe representative Chinese patent medicines included in the analysis were Shexiang Baoxin pills， Danshen tablets， Qili Qiangxin capsules， Breviscapine tablets， etc.， covering all the efficacy subcategories of "stasis-resolving agents". Among the 56 representative Chinese patent medicines， salty flavor was the most common （48）， followed by pungent （33）， and sweet （26）. According to the dominant herbal flavor， salty flavor was the most common （37）， followed by pungent （9）， and sour （5）. According to the dominant herbal quantity， salty flavor was the most common （27）， followed by sour （7）， and pungent （5）. Furthermore， Chinese patent medicines with different efficacy subtypes showed different flavor characteristics. For example， most Qi-invigorating and blood-activating agents contained sweet drugs for tonifying the spleen （9/10）， most Qi-moving and blood-activating agents contained pungent drugs for tonifying the liver （7/8）， and all kidney-invigorating and blood-activating agents contained bitter drugs for tonifying the kidneys （6/6）. However， the efficacy classification of individual medicines did not always align with the compatibility characteristics of their formulas， as seen with Dengyin Naotong capsules. ConclusionThe formulations of Chinese patent medicines for cardiovascular and cerebrovascular diseases predominantly feature salty， sour， and pungent flavors， which largely conform to the therapeutic principles of "nourishing the heart with salt and soothing the heart with sour" and the liver-heart， heart-spleen mother-child treatment relationship shown in the "Tangye Jingfa Tu". Using the "Tangye Jingfa Tu" framework to conduct research on the structure and efficacy characteristics of Chinese patent medicines is objective and effective.

ObjectiveChinese patent medicines for cardiovascular and cerebrovascular diseases are diverse and complex in their efficacy. The traditional classification method based on efficacy categories has certain limitations and cannot meet the clinical needs for individualized drug selection and variety comparison. This article， based on the formulation compatibility analysis technology of "Tangye Jingfa Tu"， clarifies the composition and efficacy characteristics of common Chinese patent medicines used for cardiovascular and cerebrovascular diseases， providing support for the precise selection of these medicines. MethodsFifty-six representative Chinese patent medicines， covering all the efficacy subcategories of "stasis-resolving agents" in the National Basic Medical Insurance， Work Injury Insurance， and Maternity Insurance Drug Catalogue （2023） （more than 50% of the total）， were selected for the study. Within the knowledge system of "Tangye Jingfa Tu"， the compatibility structure of herbal flavors and the proportion structure of herbal quantities for each Chinese patent medicine were determined. The correlation between these structures and the efficacy categories was analyzed to identify the similarities and differences among the selected Chinese patent medicines. Additionally， the efficacy was reclassified and compared according to the theoretical framework of tonifying and purging methods of five Zang organs in the "Tangye Jingfa Tu". ResultsThe representative Chinese patent medicines included in the analysis were Shexiang Baoxin pills， Danshen tablets， Qili Qiangxin capsules， Breviscapine tablets， etc.， covering all the efficacy subcategories of "stasis-resolving agents". Among the 56 representative Chinese patent medicines， salty flavor was the most common （48）， followed by pungent （33）， and sweet （26）. According to the dominant herbal flavor， salty flavor was the most common （37）， followed by pungent （9）， and sour （5）. According to the dominant herbal quantity， salty flavor was the most common （27）， followed by sour （7）， and pungent （5）. Furthermore， Chinese patent medicines with different efficacy subtypes showed different flavor characteristics. For example， most Qi-invigorating and blood-activating agents contained sweet drugs for tonifying the spleen （9/10）， most Qi-moving and blood-activating agents contained pungent drugs for tonifying the liver （7/8）， and all kidney-invigorating and blood-activating agents contained bitter drugs for tonifying the kidneys （6/6）. However， the efficacy classification of individual medicines did not always align with the compatibility characteristics of their formulas， as seen with Dengyin Naotong capsules. ConclusionThe formulations of Chinese patent medicines for cardiovascular and cerebrovascular diseases predominantly feature salty， sour， and pungent flavors， which largely conform to the therapeutic principles of "nourishing the heart with salt and soothing the heart with sour" and the liver-heart， heart-spleen mother-child treatment relationship shown in the "Tangye Jingfa Tu". Using the "Tangye Jingfa Tu" framework to conduct research on the structure and efficacy characteristics of Chinese patent medicines is objective and effective.

Background/Aims: Metabolic dysfunction-associated steatohepatitis (MASH) is a significant risk factor for gallstone formation, but mechanisms underlying MASH-related gallstone formation remain unclear. Golgi membrane protein 1 (GOLM1) participates in hepatic cholesterol metabolism and is upregulated in MASH. Here, we aimed to explore the role of GOLM1 in MASH-related gallstone formation. Methods: The UK Biobank cohort was used for etiological analysis. GOLM1 knockout (GOLM1-/-) and wild-type (WT) mice were fed with a high-fat diet (HFD). Livers were excised for histology and immunohistochemistry analysis. Gallbladders were collected to calculate incidence of cholesterol gallstones (CGSs). Biles were collected for biliary lipid analysis. HepG2 cells were used to explore underlying mechanisms. Human liver samples were used for clinical validation. Results: MASH patients had a greater risk of cholelithiasis. All HFD-fed mice developed MASH, and the incidence of gallstones was 16.7% and 75.0% in GOLM1-/- and WT mice, respectively. GOLM1-/- decreased biliary cholesterol concentration and output. In vivo and in vitro assays confirmed that GOLM1 facilitated cholesterol efflux through upregulating ATP binding cassette transporter subfamily G member 5 (ABCG5). Mechanistically, GOLM1 translocated into nucleus to promote osteopontin (OPN) transcription, thus stimulating ABCG5-mediated cholesterol efflux. Moreover, GOLM1 was upregulated by interleukin-1β (IL-1β) in a dose-dependent manner. Finally, we confirmed that IL-1β, GOLM1, OPN, and ABCG5 were enhanced in livers of MASH patients with CGSs. Conclusions In MASH livers, upregulation of GOLM1 by IL-1β increases ABCG5-mediated cholesterol efflux in an OPN-dependent manner, promoting CGS formation. GOLM1 has the potential to be a molecular hub interconnecting MASH and CGSs.

Background/Aims: Metabolic dysfunction-associated steatohepatitis (MASH) is a significant risk factor for gallstone formation, but mechanisms underlying MASH-related gallstone formation remain unclear. Golgi membrane protein 1 (GOLM1) participates in hepatic cholesterol metabolism and is upregulated in MASH. Here, we aimed to explore the role of GOLM1 in MASH-related gallstone formation. Methods: The UK Biobank cohort was used for etiological analysis. GOLM1 knockout (GOLM1-/-) and wild-type (WT) mice were fed with a high-fat diet (HFD). Livers were excised for histology and immunohistochemistry analysis. Gallbladders were collected to calculate incidence of cholesterol gallstones (CGSs). Biles were collected for biliary lipid analysis. HepG2 cells were used to explore underlying mechanisms. Human liver samples were used for clinical validation. Results: MASH patients had a greater risk of cholelithiasis. All HFD-fed mice developed MASH, and the incidence of gallstones was 16.7% and 75.0% in GOLM1-/- and WT mice, respectively. GOLM1-/- decreased biliary cholesterol concentration and output. In vivo and in vitro assays confirmed that GOLM1 facilitated cholesterol efflux through upregulating ATP binding cassette transporter subfamily G member 5 (ABCG5). Mechanistically, GOLM1 translocated into nucleus to promote osteopontin (OPN) transcription, thus stimulating ABCG5-mediated cholesterol efflux. Moreover, GOLM1 was upregulated by interleukin-1β (IL-1β) in a dose-dependent manner. Finally, we confirmed that IL-1β, GOLM1, OPN, and ABCG5 were enhanced in livers of MASH patients with CGSs. Conclusions In MASH livers, upregulation of GOLM1 by IL-1β increases ABCG5-mediated cholesterol efflux in an OPN-dependent manner, promoting CGS formation. GOLM1 has the potential to be a molecular hub interconnecting MASH and CGSs.