1.Mechanism of Modified Shaofu Zhuyutang in Antagonising Ectopic Endometrial Tissue Fibrosis Based on Cellular Pyroptosis Mediated by TRL4/NF-κB/NLPR3 Signaling Pathway
Zuoliang ZHANG ; Jiaxing WANG ; Wanrun WANG ; Xiangyu LIN ; Bin YUE ; Zhirui ZHANG ; Yinan WANG ; Yaling YANG ; Dongqing WEI ; Cancan HUANG ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):19-28
ObjectiveTo investigate the mechanism of action of modified Shaofu Zhuyutang in antagonizing cellular pyroptosis and fibrosis in ectopic endometrial tissues of endometriosis through the Toll-like receptor 4/nuclear factor-κB/NOD-like receptor protein 3 (TRL4/NF-κB/NLPR3) signaling pathway. MethodsSeventy-two SPF-grade female SD rats were randomly divided into a sham-operated group (n = 12) and a modeling group (n = 60). The rats in the sham-operated group underwent a caesarean section, while the rats in the modeling group were used to establish an endometriosis model through the auto-transplantation method. After successful modeling, the animals were randomly divided into the model group, progesterone group (0.25 mg·kg-1), and modified Shaofu Zhuyutang low-, medium-, and high-dose groups (7.5, 15, 30 g·kg-1), with 12 animals in each group. After 4 weeks of drug administration, voluntary activity and heat pain latency were observed. The rats were sacrificed for tissue collection, and Masson staining were used to observe histopathological changes in the endometrial tissues. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of interleukin-18 (IL-18), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and transforming growth factor-β (TGF-β). Immunohistochemistry (IHC) was used to detect the protein expression area of tumor necrosis factor-related factor 6 (TRAF6) and NLPR3 in the endometrial tissues. Immunofluorescence (IF) was used to detect the relative fluorescence intensity of Caspase-1 and gasdermin D (GSDMD) in the endometrial tissues. Western blot was employed to measure the relative expression of TRL4, myeloid differentiation factor 88 (MyD88), TRAF6, NF-κB p65, phosphorylated NF-κB p65 (p-NF-κB p65), and NLPR3 proteins in endometrial tissues. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of TRL4, MyD88, TRAF6, NF-κB, and NLPR3 in the endometrial tissues. ResultsCompared with the sham-operated group, rats in the model group showed reduced voluntary activity and shorter heat pain latency. Serum levels of IL-18, IL-1β, TNF-α, and TGF-β were elevated. The relative expression areas of TRAF6 and NLPR3 proteins were increased, and the relative fluorescence intensity of Caspase-1 and GSDMD was enhanced. The relative expression of TRL4, MyD88, TRAF6, NF-κB p65, p-NF-κB p65, and NLPR3 proteins, along with the expression of TRL4, MyD88, TRAF6, NF-κB, and NLPR3 mRNA, were significantly increased (P<0.01). Compared with the model group, rats in the progesterone group and the modified Shaofu Zhuyutang medium- and high-dose groups exhibited improved voluntary activity, longer heat pain latency, the fibrosis of endometrial tissue is alleviated. Serum levels of IL-18, IL-1β, TNF-α, and TGF-β were decreased. The relative expression areas of TRAF6 and NLPR3 proteins decreased, and the relative fluorescence intensity of Caspase-1 and GSDMD weakened. The relative expression of TRL4, MyD88, TRAF6, p-NF-κB p65, NLPR3 proteins, and TRL4, MyD88, TRAF6, NF-κB, and NLPR3 mRNA expression were reduced (P<0.05, P<0.01). ConclusionModified Shaofu Zhuyutang may play a therapeutic role in endometriosis by interfering with key proteins in the TRL4/NF-κB/NLPR3 signaling pathway, reducing NLRP3 inflammasome-induced cellular pyroptosis, antagonizing the fibrosis process in ectopic endometrial tissues, improving the inflammatory microenvironment in the pelvic cavity, and alleviating pain.
2.Mechanism of Modified Shaofu Zhuyutang in Treatment of Endometriosis Based on EGFR/PI3K/Akt Signaling Pathway
Yaling YANG ; Wanrun WANG ; Zuoliang ZHANG ; Xiangyu LIN ; Jiaxing WANG ; Cancan HUANG ; Xiujia JI ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):29-38
ObjectiveTo observe the effects of modified Shaofu Zhuyutang on key proteins of the epidermal growth factor receptor (EGFR)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in SD rats with endometriosis. MethodsAfter successful establishment of an endometriosis model in 60 female SD rats of SPF grade via the auto-transplantation method, the rats were randomly divided into a model group, modified Shaofu Zhuyutang high-, medium-, and low-dose groups, and a gestrinone group, with another 12 rats serving as a blank group. The blank and model groups were administered 10 mL·kg-1 normal saline, while the high-, medium-, and low-dose groups received 30, 15, and 7.5 g·kg-1 modified Shaofu Zhuyutang, respectively. The gestrinone group was administered 0.25 mg·kg-1 gestrinone suspension. After four weeks of treatment, uterine contractions were induced with 2 U of oxytocin, and the writhing response of rats was observed. After 24 h, the rats were euthanized, and the weight and volume of ectopic endometrial tissue were recorded. Hematoxylin-eosin (HE) staining was used to observe pathological changes in endometrial tissues, while the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay was used to evaluate the apoptosis rate of endometrial tissues. Immunofluorescence was used to detect the relative expression areas of the B-cell lymphoma-2 gene-associated promoter (Bad) and B-cell lymphoma-2 (Bcl-2) proteins in endometrial tissues. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), epidermal growth factor (EGF), and EGFR were measured by enzyme-linked immunosorbent assay (ELISA). The relative protein expression levels of EGFR, PI3K, phosphorylated PI3K (p-PI3K), Akt, and phosphorylated Akt (p-Akt) in endometrial tissues were analyzed by Western blot. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of EGFR, PI3K, and Akt. ResultsCompared with the blank group, the model group showed endometrial thickening, glandular and mesenchymal hyperplasia, a significant decrease in the relative expression area of Bad in ectopic endometrial tissues, a significant increase in the relative expression area of Bcl-2, and a significant reduction in the apoptosis rate as indicated by TUNEL staining. Serum levels of IL-1β, IL-6, TNF-α, EGF, and EGFR were significantly elevated (P<0.01). The relative protein expression levels of EGFR, PI3K, p-PI3K, Akt, and p-Akt, as well as the mRNA expression levels of EGFR, PI3K, and Akt, were also significantly increased (P<0.01). Compared with the model group, the high- and medium-dose groups of modified Shaofu Zhuyutang and the gestrinone group exhibited reduced glandular and mesenchymal hyperplasia to varying degrees, with dilated glandular lumens. The number of writhing responses was significantly reduced, the latency to writhing response was significantly prolonged, and the weight and volume of ectopic endometrial tissue were significantly decreased. The relative expression area of Bad in ectopic endometrial tissue was significantly increased, the relative expression area of Bcl-2 was significantly decreased, and the apoptosis rate was significantly elevated as shown by TUNEL staining. Serum levels of IL-1β, IL-6, TNF-α, EGF, and EGFR were significantly reduced, and the relative protein expression levels of EGFR, PI3K, p-PI3K, Akt, and p-Akt, as well as the mRNA expression levels of EGFR, PI3K, and Akt, were significantly decreased (P<0.05,P<0.01). ConclusionModified Shaofu Zhuyutang may exert therapeutic effects on endometriosis by interfering with key proteins of the EGFR/PI3K/Akt signaling pathway and inducing apoptosis in ectopic endometrial tissue.
3.Effect of Modified Shaofu Zhuyutang on Ferroptosis in Ectopic Endometrial Tissues of Rats with Endometriosis Based on MDM4/p53/GPX4 Signaling Pathway
Zuoliang ZHANG ; Xiangyu LIN ; Wanrun WANG ; Jiaxing WANG ; Yaling YANG ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):39-47
ObjectiveTo investigate the mechanism of modified Shaofu Zhuyutang in inducing ferroptosis in ectopic endometrial tissues of rats with endometriosis through the murine double minute 4 (MDM4)/tumor suppressor p53/glutathione peroxidase 4 (GPX4) signaling pathway. MethodsSeventy SPF-grade female SD rats were randomly divided into a blank group (n = 10), a sham-operated group (n = 10), and a modeling group (n = 50). The sham-operated group underwent laparotomy, while the modeling group was subjected to the autotransplantation method to establish an endometriosis model. After successful modeling, the animals were randomly assigned to the model group, progesterone group (0.25 mg·kg-1), and modified Shaofu Zhuyutang high-, medium-, and low-dose groups (30, 15, and 7.5 g·kg-1, respectively), with 10 rats per group. After four weeks of drug administration, the rats were euthanized for sample collection. The weight and volume of ectopic endometrial tissues were recorded for each group. Transmission electron microscopy (TEM) was employed to observe ultrastructural changes in endometrial tissues, while Prussian blue staining was used to assess iron ion deposition. Serum levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). The relative levels of Fe2+, malondialdehyde (MDA), and glutathione (GSH) in endometrial tissues were determined by colorimetric assay. Immunofluorescence (IF) was used to detect the relative fluorescence intensities of GSH and GPX4 in endometrial tissues. The relative expression levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K (p-PI3K), protein kinase B (Akt), phosphorylated Akt (p-Akt), MDM4, p53, and GPX4 proteins were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to assess the mRNA expression of PI3K, Akt, MDM4, p53, and GPX4. ResultsCompared with the blank and sham-operated groups, the model group exhibited reduced ferroptotic damage in ultrastructural observations, decreased ferroptotic aggregates and positive iron ion expression area on Prussian blue staining, elevated serum IL-6, IL-1β, and TNF-α levels, reduced Fe2+ and MDA content, increased GSH content in endometrial tissues, and enhanced GSH and GPX4 fluorescence intensities (P<0.01). The protein and mRNA expression levels of PI3K, p-PI3K, Akt, p-Akt, MDM4, and GPX4 were elevated, while those of p53 were decreased (P<0.01). Compared with the model group, in the progesterone group and the modified Shaofu Zhuyutang high- and medium-dose groups, ferroptotic damage in ultrastructural observations was exacerbated to varying degrees by TEM, and ferroptotic aggregates and positive iron ion expression areas were increased on Prussian blue staining. Serum IL-6, IL-1β, and TNF-α levels decreased, Fe2+ and MDA content increased, and GSH content decreased in endometrial tissues. GSH and GPX4 fluorescence intensities weakened, while the protein and mRNA expression levels of PI3K, p-PI3K, Akt, p-Akt, MDM4, and GPX4 decreased, and those of p53 increased (P<0.05, P<0.01). Conclusionmodified Shaofu Zhuyutang may exert therapeutic effects in endometriosis by inducing ferroptosis in ectopic endometrial tissues, alleviating inflammatory responses, and modulating key proteins in the MDM4/p53/GPX4 signaling pathway.
4.Comparison of multiple machine learning models for predicting the survival of recipients after lung transplantation
Lingzhi SHI ; Yaling LIU ; Haoji YAN ; Zengwei YU ; Senlin HOU ; Mingzhao LIU ; Hang YANG ; Bo WU ; Dong TIAN ; Jingyu CHEN
Organ Transplantation 2025;16(2):264-271
Objective To compare the performance and efficacy of prognostic models constructed by different machine learning algorithms in predicting the survival period of lung transplantation (LTx) recipients. Methods Data from 483 recipients who underwent LTx were retrospectively collected. All recipients were divided into a training set and a validation set at a ratio of 7:3. The 24 collected variables were screened based on variable importance (VIMP). Prognostic models were constructed using random survival forest (RSF) and extreme gradient boosting tree (XGBoost). The performance of the models was evaluated using the integrated area under the curve (iAUC) and time-dependent area under the curve (tAUC). Results There were no significant statistical differences in the variables between the training set and the validation set. The top 15 variables ranked by VIMP were used for modeling and the length of stay in the intensive care unit (ICU) was determined as the most important factor. Compared with the XGBoost model, the RSF model demonstrated better performance in predicting the survival period of recipients (iAUC 0.773 vs. 0.723). The RSF model also showed better performance in predicting the 6-month survival period (tAUC 6 months 0.884 vs. 0.809, P = 0.009) and 1-year survival period (tAUC 1 year 0.896 vs. 0.825, P = 0.013) of recipients. Based on the prediction cut-off values of the two algorithms, LTx recipients were divided into high-risk and low-risk groups. The survival analysis results of both models showed that the survival rate of recipients in the high-risk group was significantly lower than that in the low-risk group (P<0.001). Conclusions Compared with XGBoost, the machine learning prognostic model developed based on the RSF algorithm may preferably predict the survival period of LTx recipients.
5.Study on stir-frying process of Platycodon grandiflorum and its protective effect on acute lung injury in mice before and after stir-frying
Xiaotian HAN ; Lei WANG ; Yaping WANG ; Yaling YANG ; Bin QI
China Pharmacy 2025;36(13):1587-1592
OBJECTIVE To optimize the stir-frying process of Platycodon grandiflorum, compare the protective effect of P. grandiflorum on lung injury in mice before and after stir-frying, and preliminarily explore the characteristics of “enhanced efficacy through processing” of stir-frying P. grandiflorum. METHODS On the basis of single-factor experiment, the stir-frying time, temperature and frequency were taken as factors, and the comprehensive scores of appearance traits, platycodin D content and alcohol extract content were taken as indexes. Box-Behnken response surface experiment was designed to optimize the stir-frying process of P. grandiflorum and verify it. The mice were divided into blank control group, model group, dexamethasone group (positive control drug, 5 mg/kg), P. grandiflorum low-dose and high-dose groups (0.6,1.2 g/kg), fried P. grandiflorum low and high dose groups (0.6, 1.2 g/kg), with 8 mice in each group. The treatment was given once a day for 10 consecutive days. After the last administration, acute lung injury model was established by intratracheal instillation of lipopolysaccharide. The levels of interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α) in serum and superoxide dismutase (SOD), myeloperoxidase (MPO) and malondialdehyde (MDA) in lung tissue were detected, lung wet/dry weight ratio and thymus index were calculated, and the pathological changes of lung tissue were observed. RESULTS The optimum processing conditions were as follows:stir- frying temperature of 120 ℃, stir-frying time of 12 min, stir-frying frequency of 19 r/min; the comprehensive scores of the three batches of process verification were all greater than 97 points, RSD<3% (n=3). The results of pharmacodynamic experiments showed that compared with blank control group, the lung dry-wet weight ratio as well as the levels of IL-6, IL-1β, TNF-α, MPO and MDA in the model group were significantly increased (P<0.01), the thymus index and SOD level were significantly decreased (P<0.01), and the lung tissue was significantly damaged. Compared with model group, above indexes of the mice in each administration group were significantly improved (P<0.01), and the lung tissue injury was significantly reduced. At the same dose, except for the lung dry-wet weight ratio, the above indexes of the mice in the stir-fried P. grandiflorum groups were significantly improved compared with P. grandiflorum 涵。E-mail:495758271@qq.com groups (P<0.05), and the lung tissue damage was further reduced. CONCLUSIONS The optimized stir-frying process is stable and feasible. The protective effect of stir-fried P.grandiflorum on acute lung injury in mice is better than that of raw products at the same dosage.
6.Application and data analysis of the cardio-cerebrovascular events monitoring system in Yichang
Zhengchao FANG ; Jiajuan YANG ; Chi HU ; Chan WU ; Yaling DENG ; Zhiying YU ; Jie ZHU ; Ling ZHANG
Journal of Public Health and Preventive Medicine 2025;36(6):95-98
Objective To analyze the monitoring data of cardio-cerebrovascular diseases prevention and control system in Yichang in 2022, and to provide data support and experience for the precise prevention and treatment of cardio-cerebrovascular diseases. Methods Acute cardiovascular and cerebrovascular event data were collected from the Yichang Cardio-cerebrovascular Events Monitoring System from January 1, 2022 to December 31, 2022. Descriptive analysis was conducted for the data collected. Statistical analysis was performed using SPSS 20.0 software, and a chi-square test was used to analyze the count data. Results A total of 37,217 cases of cardio-cerebrovascular events were monitored in Yichang in 2022. The crude incidence and the standardized incidence were 983.84/100,000 and 541.55/100,000, respectively. The incidence in males was higher than females (554.93/100,000 vs 428.91/100,000,χ2 =464.52,P<0.05). The top three diseases were cerebral infarction, acute myocardial infarction, and cerebral hemorrhage. The incidence of events increased with age, and 79.80% of the cases were over 60 years old. The main onset time was from May to August. Conclusion The use of the cardio-cerebrovascular events monitoring system in Yichang and the implementation of “mandatory reporting card” monitoring can timely obtain the epidemic characteristics of the diseases, provide support for the precise formulation of prevention and control strategies and measures, reduce underreporting rates, and improve the monitoring system, which is worthy of reference and promotion.
7.Evaluation and improvement of undergraduate courses in food hygiene and nutrition from the perspective of graduates
Qian ZHANG ; Yan YANG ; Yaling QIE ; Hongwei SU ; Ling MA
Chinese Journal of Medical Education Research 2024;23(7):926-930
Objective:To understand the problems existing in the curriculum design of food hygiene and nutrition for undergraduates, and put forward countermeasures to improve the quality of talents cultivation and promote professional development.Methods:Self-made questionnaires were distributed by convenience sampling method, through online and offline ways, and the graduates' opinions on the curriculum design of their school. Quantitative data were analyzed by SPSS 26.0, and qualitative data were analyzed by NVivo 11.0.Results:A total of 258 valid questionnaires were recovered, and the average score of graduates' satisfaction with teaching quality was 2.42±1.02. Graduates had different opinions on the importance of special courses, and 150 students (58.14%) were satisfied with major courses. Some graduates thought the time of special courses was unreasonable and there were few major courses. Besides, 141 graduates (54.65%) were satisfied with the clinical probation in their graduation colleges. Although graduates believed that innovation ability was important, innovative education courses were scarce at present.Conclusions:Graduates are less satisfied with the setting of undergraduate curriculum in food hygiene and nutrition. It is suggested to optimize professional courses setting, improve experimental facilities and equipment, strengthen the management of practice bases, and pay attention to the cultivation of students' innovation and entrepreneurship ability.
8.Targeted delivery of rosuvastatin enhances treatment of hyperhomocysteinemia-induced atherosclerosis using macrophage membrane-coated nanoparticles
Liu DAYUE ; Yang ANNING ; Li YULIN ; Li ZHENXIAN ; You PEIDONG ; Zhang HONGWEN ; Quan SHANGKUN ; Sun YUE ; Zeng YALING ; Ma SHENGCHAO ; Xiong JIANTUAN ; Hao YINJU ; Li GUIZHONG ; Liu BIN ; Zhang HUIPING ; Jiang YIDENG
Journal of Pharmaceutical Analysis 2024;14(9):1301-1319
Rosuvastatin(RVS)is an excellent drug with anti-inflammatory and lipid-lowering properties in the aca-demic and medical fields.However,this drug faces a series of challenges when used to treat atherosclerosis caused by hyperhomocysteinemia(HHcy),including high oral dosage,poor targeting,and long-term toxic side effects.In this study,we applied nanotechnology to construct a biomimetic nano-delivery system,macrophage membrane(M?m)-coated RVS-loaded Prussian blue(PB)nanoparticles(MPR NPs),for improving the bioavailability and targeting capacity of RVS,specifically to the plaque lesions associated with HHcy-induced atherosclerosis.In vitro assays demonstrated that MPR NPs effectively inhibited the Toll-like receptor 4(TLR4)/hypoxia-inducible factor-1α(HIF-1α)/nucleotide-binding and oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathways,reducing pyroptosis and inflammatory response in macrophages.Additionally,MPR NPs reversed the abnormal distribution of adenosine triphosphate(ATP)-binding cassette transporter A1(ABCA1)/ATP binding cassette transporter G1(ABCA1)/ATP binding cassette transporter G1(ABCG1)caused by HIF-1α,promoting cholesterol efflux and reducing lipid deposition.In vivo studies using apolipoprotein E knockout(ApoE-/-)mice confirmed the strong efficacy of MPR NPs in treating atherosclerosis with favorable bio-security,and the mechanism behind this efficacy is believed to involve the regulation of serum metabolism and the remodeling of gut microbes.These findings suggest that the synthesis of MPR NPs provides a promising nanosystem for the targeted therapy of HHcy-induced atherosclerosis.
9.Research progress of macrophage based nanoimaging and nanobiomimetic technology in the diagnosis and treatment of atherosclerotic plaque
Yaling WU ; Yuanhui DUAN ; Rong SUN ; Zhizhen SHI ; Lijia YANG ; Sen QUN
Chinese Journal of Arteriosclerosis 2024;32(10):899-906
Macrophages play an important role in the development of inflammation,the core mechanism of athero-sclerosis.In recent years,nanotechnology has provided a new perspective for the diagnosis and treatment of atherosclerosis with its unique characteristics of biological compatibility and precise targeting.In this paper,the patholog-ical mechanism of atherosclerosis,the realization of atherosclerosis imaging by targeting macrophages with nanomaterials and the application of macrophage cell membrane biomimetic nanoparticles in anti-atherosclerosis were reviewed.
10.Clinical analysis of hematopoietic stem cell transplantation for the treatment of eld-erly myeloid tumors
Zheng YALING ; Zheng ZIYU ; Wu MING ; Yang CHUTING ; Huang RUI ; Li YUHUA
Chinese Journal of Clinical Oncology 2024;51(11):552-556
Objective:To investigate the effect of hematopoietic stem cell transplantationon(HSCT)survival outcomes in older patients with myeloid neoplasms.Methods:We retrospectively analyzed the treatment outcomes of 54 patients aged≥55 years with myeloid neoplasms who underwent HSCT between January 2018 and May 2023 at Zhujiang Hospital of Southern Medical University.Results:Among the 54 pa-tients,45 had acute myeloid leukemia(AML)and 9 had myelodysplastic syndrome.The median age of the patients was 57.5(55-68)years.Fifty-three patients underwent hematopoietic reconstitution,with a median time to neutrophil reconstitution of 13(8-24)days and median time to platelet reconstitution of 15(9-75)days.The cumulative incidence was 23.3%for acute graft-versus-host disease(GVHD)and 24.6%for 3-year chronic GVHD.With a median follow-up of 28.2 months,the 3-year cumulative relapse rate(CIR)was 18%and 3-year non-relapse mortality rate was 28.3%.The 3-year relapse-free survival(RFS)rate was 58.2%and 3-year overall survival(OS)rate was 56.5%.Conclu-sions:HSCT is an effective and safe therapy for achieving long-term survival in older patients with myeloid tumors.


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