ObjectiveThis study aims to investigate the molecular mechanism by which Guizhi Fulingwan （GFW） inhibits ferroptosis in endometriosis （EMT） through the regulation of the hypoxia inducible factor-1α/heme oxygenase 1 （HIF-1α/HO-1） signaling pathway. MethodsMachine learning was employed to identify ferroptosis-related biomarkers associated with EMT. Network pharmacology was utilized to identify the active components of GFW and its potential therapeutic targets against EMT， including core targets. Functional enrichment analysis was conducted to explore the biological processes， molecular functions， cellular components， and signaling pathways associated with the potential targets. An EMT rat model was established via autologous transplantation. Thirty female Sprague-Dawley （SD） rats were randomly divided into five groups： sham-operated， model， positive control （dienogest at 0.2 mg·kg^-1）， low-dose GFW （2.5 g·kg^-1）， and high-dose GFW （5 g·kg^-1）. After modeling， the rats received their respective treatment by oral gavage for 28 consecutive days， while the sham and model groups received equal volumes of distilled water. Serum and ectopic endometrial tissues were collected. Hematoxylin and eosin （HE） staining was employed to evaluate morphological alterations in ectopic lesions. Quantitative real-time polymerase chain reaction （Real-time PCR） and Western blot were conducted to assess mRNA and protein expression of HIF-1α， HO-1， glutathione peroxidase 4 （GPX4）， spermidine/spermine N1-acetyltransferase （SAT1）， and prostaglandin-endoperoxide synthase 2 （PTGS2）. Tissue levels of malondialdehyde （MDA）， glutathione （GSH）， and ferrous iron （Fe²⁺） were quantified using commercial assay kits. Serum levels of interleukin-6 （IL-6） and transforming growth factor-β₁ （TGF-β₁） were measured via enzyme-linked immunosorbent assay （ELISA）. ResultsFive ferroptosis-related biomarkers in EMT were identified： ALOX12， CHAC1， SAT1， AST1， and HO-1. Network pharmacology analysis revealed 42 active components of GFW and 192 potential therapeutic target genes related to EMT treatment， with FOS， JUN， HO-1 identified as core targets. Functional enrichment analysis indicated that the potential targets were primarily involved in oxidative stress response and reactive oxygen species metabolism and were enriched in the HIF-1 signaling pathway. Compared to the sham-operated group， the model group exhibited significant increases in both mRNA and protein expression of HIF-1α， HO-1， and PTGS2， as well as elevated tissue levels of Fe²⁺ and MDA. Conversely， GSH levels and the expression of GPX4 and SAT1 were markedly reduced， and serum levels of IL-6 and TGF-β₁ levels were significantly higher （P<0.01）. Compared with the model group， all GFW-treated groups showed significant downregulation of HIF-1α and HO-1， reduced Fe²⁺ levels， and downregulated expression of MDA， PTGS2， IL-6， and TGF-β₁. Meanwhile， GSH， GPX4， and SAT1 expression levels were significantly increased （P<0.05， P<0.01）， effectively ameliorating iron overload and oxidative stress， thereby demonstrating therapeutic efficacy in EMT， with the high-dose GFW demonstrating the most pronounced therapeutic effects. ConclusionGFW exerts therapeutic effects on endometriosis by regulating the HIF-1α/HO-1 signaling pathway to rectify iron metabolism disorders and attenuate free iron-induced oxidative damage. It upregulates the antioxidative defense system to inhibit lipid peroxidation cascades and modulates inflammatory cytokine networks. These effects collectively disrupt the pathological interaction between ferroptosis and chronic inflammation， providing a novel theoretical foundation for the clinical application of GFW in EMT treatment.

Objective To investigate the effects of Echinococcus multilocularis infection on levels of memory T (Tm) cells and their subsets in lymph nodes of mice at different stages of infection, so as to provide new insights into immunotherapy for alveolarechinococcosis. MethodsTwenty-four C57BL/6J mice aged 6 to 9 weeks were randomly divided into the infection group and the control group, of 12 mice in each group. Mice in the infection group were administered with 3 000 E. multilocularis protoscoleces via portal venous injection, while animals in the control group were administered with an equal volume of physiological saline. Three mice from each group were sacrificed 4, 12 weeks and 24 weeks post-infection, and lymph nodes were sampled and stained with hematoxylin and eosin (HE) to investigate the histopathological changes of mouse lymph nodes in the infection group. The expression and localization of T lymphocyte surface markers CD3, CD4, and CD8 were observed in mouse lymph nodes using immunohistochemical staining. In addition, lymphocyte suspensions were prepared from mouse lymph nodes in both groups at different time points post-infection, and the levels of Tm cell subsets and their secreted cytokines were detected using flow cytometry. Results HE staining showed diffuse structural alterations in the subcapsular cortical and paracortical regions of mouse lymph nodes in the infection group 4 weeks post-infection with E. multilocularis. Immunohistochemical staining detected CD3, CD4 and CD8 expression in mouse lymph nodes in both groups. Flow cytometry revealed higher proportions of CD4⁺ Tm cells [(55.3 ± 4.8)% vs. (38.8 ± 6.1)%; t = -4.259, P < 0.05] and CD4⁺ tissue-resident Tm (Trm) cells [(57.7 ± 3.7)% vs. (34.1 ± 11.2)%; t = -3.990, P < 0.05] in mouse lymph nodes in the infection group than in the control group 4 weeks post-infection, and higher proportions of CD4⁺ Tm cells [(34.6 ± 3.2)% vs. (23.3 ± 7.5)%; t = -2.764, P < 0.05] and CD4⁺ Trm cells [(44.0 ± 1.9)% vs. (31.2 ± 1.5)%; t = -4.039, P < 0.05] in mouse lymph nodes in the infection group than in the control group 24 weeks post-infection. The proportions of CD8⁺ Tm cells were higher in the infection group than in the control group 4 weeks [(56.8 ± 2.7)% vs. (43.9 ± 5.2)%; t = -4.416, P < 0.01] and 12 weeks post-infection [(25.4 ± 2.7)% vs. (12.0 ± 2.6)%; t = -2.552, P < 0.05], while the proportions of tumor necrosis factor (TNF)-α⁺ CD4⁺ T cells [(15.7 ± 5.0)% vs. (49.4 ± 6.4)%; t = 7.150, P < 0.01], TNF-α⁺CD8⁺ T cells [(20.7 ± 5.5)% vs. (57.5 ± 8.4)%; t = -6.694, P < 0.01], and TNF-α⁺ CD8⁺ Tm cells [7.0% (1.0%) vs. 31.0% (11.0%); Z = -2.236, P < 0.05] were lower in the infection group than in the control group 24 weeks post-infection. Conclusions Tm cells levels are consistently increased in lymph nodes of mice at different stages of E. multilocularis infection, with Trm cells as the predominantly elevated subset. The impaired capacity of CD8⁺ Tm cells to secrete the effector molecule TNF-α in mouse lymph nodes at the late-stage infection may facilitate chronic parasitism of E. multilocularis.

Objective To investigate the multi-target mechanisms of quercetin in treating endometriosis(EMT)through integrative network pharmacology analysis.Methods Active targets of quercetin were collected from the TCMSP database,while EMT-related differentially expressed genes(DEGs)were identified through the Gene Expression Omnibus(GEO)dataset.A comparative analysis was conducted to pinpoint potential therapeutic targets of quercetin for EMT treatment.Functional enrichment analyses were employed to investigate the biological functions associated with these targets,and a protein-protein interaction(PPI)network was conducted to identify core targets.Molecular docking and dynamics simulations were performed to validate the binding characteristics between quercetin and the core targets.The top 2 target protein pairs,HSP90AB1 and AR,exhibiting the lowest binding energy,were selected for subsequent cellular experimental validation.Human EMT-immortalized ectopic endometrial epithelial cell line 12Z(n=6,independent replicates)was subjected,and CCK-8 assay was used to determine ehe effects of quercetin on cell viability and proliferation,and the half-maximal inhibitory concentration(IC50)was calculated at 48 h after treatment.Then the 12Z cells were treated with quercetin at a concentration gradient of 0,30,60 and 90 μmol/L,the migration and invasion abilities were assessed with cell scratch and cell invasion assays.Western blotting was conducted to detect the changes in the expression of HSP90AB1 and AR proteins after different doses of treatment.Results There were 49 potential EMT-related therapeutic targets and 10 core targets identified.Functional enrichment analyses revealed that these targets were significant enriched in inflammation-related signaling pathways,including AGE-RAGE,ErbB and TNF;immune-related pathways,such as Th17 cell differentiation,T/B cell receptor signaling;angiogenesis-related pathways like VEGF;and hormonal regulatory pathways involving estrogen and GnRH.Molecular docking demonstrated that quercetin exhibited favorable binding activity(binding energy<-5 kcal/mol)with all core target proteins,with particularly strong binding energies(<-7 kcal/mol)observed for AR,EGFR,FOS,ERBB2,and HSP90AB1.Molecular dynamics simulations revealed that quercetin forms sustained hydrogen bond interactions with AR and HSP90AB1,facilitating the formation of stable complexes.CCK-8 assay,cell scratch assay,and transwell invasion assay indicated that quercetin inhibited the proliferative activity,and migrative and invasive abilities of 12Z cells in a concentration-dependent manner,with more pronounced inhibitory effects observed at 60 and 90 μmol/L quercetin(P<0.001);Western blotting revealed that treatment of 12Z cells with varying quercetin concentrations for 48 h up-regulated the expression of HSP90AB1 and AR,with the most significant increase observed at 90 μmol/L quercetin(HSP90AB1,P<0.05;AR,P<0.001).The restored expression levels of HSP90AB1 and AR showed positive correlations with the proliferative activity,migrative and invasive abilities of ectopic endometrial cells.Conclusion Quercetin effectively addresses endometriosis through multiple molecular targets and signaling pathways,and stabilization of the HSP90AB1/AR complex and subsequent protein upregulation represents a key therapeutic mechanism.

Retinopathy caused by myopia is the first cause of irreversible blinding eye disease in China.The TCM methods to prevent and control myopia mainly include Chinese materia medica and TCM appropriate techniques,which have the advantages of good efficacy,simple operation,and few adverse reactions.This paper believed that internal and external pathogenic factors act on the brain and eyes,breaking their homeostasis,leading to rhythmic disorders,and imbalance of essence,qi and blood is the main pathogenesis of myopia.Based on the idea of"nature and human in the same rhythm"and"the same treatment for common diseases",targeting the above pathogenesis,the method of"brain-eye synchronization"was proposed to restore the homeostasis of the brain and eyes to prevent and control myopia in children and adolescents,and the treatment rules were to regulate the rhythms of the time,harmonize the qi and blood,nourish the blood to soften the tendons,and replenish the essence and blood,so as to achieve the effect of brain-eye synchronization and the treatment of the spirit and the body together.This article summarized the theoretical basis of"brain-eye synchronization"and its clinical application in traditional Chinese and Western medicine,with a view to providing new ideas for the prevention and control of myopia in children and adolescents.

Objective:To explore risk factors for delayed onset of lactogenesis in prenatally overweight women and construct risk prediction models based on machine learning for early identification of high-risk individuals.Methods:Convenience sampling was adopted to select 338 prenatally overweight women who delivered in the Obstetrics Departments of four ClassⅢ Grade A hospitals in Hengyang City from October 2023 to June 2024 for the study. Delivery women were randomly divided into training set and test set in the ratio of 7∶3. The survey was conducted with the General Information Questionnaire, Breastfeeding Self-efficacy Scale Short Form, Edinburgh Postnatal Depression Scale, LATCH Scale and Pittsburgh Sleep Quality Index. One-way analysis and LASSO regression were used to screen predictors using delayed onset of lactogenesis as the outcome variable. Risk prediction models were constructed based on three machine learning algorithms of Logistic regression, support vector, and random forest, respectively. The models were tuned by ten-fold cross-validation to filter out the best models.Results:Delayed onset of lactogenesis occurred in 140 of 338 prenatally overweight women, an incidence of 41.4%. Among the three predictive model performances, the random forest model had the highest area under the receiver operating characteristic curve, accuracy, precision, recall, and F1 value. The importance of each predictor was ranked according to the fandom forest algorithm, and in descending order of importance, they were breastfeeding 1 h after the birth of the newborn, number of previous deliveries, age, feeding mode 3 d postpartum, pregnancy complications, mode of delivery, number of breastfeeding 24 h postpartum, and monthly household income.Conclusions:Risk prediction models for delayed onset of lactogenesis in prenatally overweight women are constructed based on three machine learning algorithms, aiming to help provide a scientific basis for clinical healthcare professionals to take relevant decisions.

Traditional Chinese medicine upholds the unity of nature and humanity as its fundamental principle,asserting that man is in harmony with heaven and earth and corresponds to the sun and moon.Human life processes align with the circadian rhythms of nature,exhibiting periodic temporal changes-concepts that Western medicine defines as biological rhythms.The"opening-closing-pivot"of the three yin and three yang meridians represents the primary mode of interaction between nature and humanity.This mechanism regulates the transformation of yin and yang to qi within the body,ensuring synchronization with the circadian rhythm.Building upon this foundation,the concept of"the harmony of rhythms and states"was proposed to explain human growth and aging."Rhythms"encompass not only circadian cycles but also the movement of qi and blood,as well as the contraction rhythm of muscle fascia,among other physiological processes."States"refer to the dynamic evolution and transformation of qi during the"opening-closing-pivot"stages in both time and space."Harmony"pertains not only to the individual physiological balance of"rhythms"or"states"but also to their coherent interaction.The fundamental cause of myopia is"the disharmony of rhythms and states",considering that rhythm disorders,qi and blood disorders,and fascia imbalances contribute to this disease.Based on this understanding,myopia prevention and treatment should center on"regulating rhythms and states",with adjusting the pivot to brighten the eyes as the core therapeutic principle throughout the entire cycle of myopia.Treatment should be tailored to different disease stages and supplemented with methods such as time-based regulation,qi and blood harmonization,tendon relaxation and collateral dredging,and essence and blood nourishment to prevent and manage myopia effectively.

Retinopathy caused by myopia is the first cause of irreversible blinding eye disease in China.The TCM methods to prevent and control myopia mainly include Chinese materia medica and TCM appropriate techniques,which have the advantages of good efficacy,simple operation,and few adverse reactions.This paper believed that internal and external pathogenic factors act on the brain and eyes,breaking their homeostasis,leading to rhythmic disorders,and imbalance of essence,qi and blood is the main pathogenesis of myopia.Based on the idea of"nature and human in the same rhythm"and"the same treatment for common diseases",targeting the above pathogenesis,the method of"brain-eye synchronization"was proposed to restore the homeostasis of the brain and eyes to prevent and control myopia in children and adolescents,and the treatment rules were to regulate the rhythms of the time,harmonize the qi and blood,nourish the blood to soften the tendons,and replenish the essence and blood,so as to achieve the effect of brain-eye synchronization and the treatment of the spirit and the body together.This article summarized the theoretical basis of"brain-eye synchronization"and its clinical application in traditional Chinese and Western medicine,with a view to providing new ideas for the prevention and control of myopia in children and adolescents.

Traditional Chinese medicine upholds the unity of nature and humanity as its fundamental principle,asserting that man is in harmony with heaven and earth and corresponds to the sun and moon.Human life processes align with the circadian rhythms of nature,exhibiting periodic temporal changes-concepts that Western medicine defines as biological rhythms.The"opening-closing-pivot"of the three yin and three yang meridians represents the primary mode of interaction between nature and humanity.This mechanism regulates the transformation of yin and yang to qi within the body,ensuring synchronization with the circadian rhythm.Building upon this foundation,the concept of"the harmony of rhythms and states"was proposed to explain human growth and aging."Rhythms"encompass not only circadian cycles but also the movement of qi and blood,as well as the contraction rhythm of muscle fascia,among other physiological processes."States"refer to the dynamic evolution and transformation of qi during the"opening-closing-pivot"stages in both time and space."Harmony"pertains not only to the individual physiological balance of"rhythms"or"states"but also to their coherent interaction.The fundamental cause of myopia is"the disharmony of rhythms and states",considering that rhythm disorders,qi and blood disorders,and fascia imbalances contribute to this disease.Based on this understanding,myopia prevention and treatment should center on"regulating rhythms and states",with adjusting the pivot to brighten the eyes as the core therapeutic principle throughout the entire cycle of myopia.Treatment should be tailored to different disease stages and supplemented with methods such as time-based regulation,qi and blood harmonization,tendon relaxation and collateral dredging,and essence and blood nourishment to prevent and manage myopia effectively.

Objective:To explore risk factors for delayed onset of lactogenesis in prenatally overweight women and construct risk prediction models based on machine learning for early identification of high-risk individuals.Methods:Convenience sampling was adopted to select 338 prenatally overweight women who delivered in the Obstetrics Departments of four ClassⅢ Grade A hospitals in Hengyang City from October 2023 to June 2024 for the study. Delivery women were randomly divided into training set and test set in the ratio of 7∶3. The survey was conducted with the General Information Questionnaire, Breastfeeding Self-efficacy Scale Short Form, Edinburgh Postnatal Depression Scale, LATCH Scale and Pittsburgh Sleep Quality Index. One-way analysis and LASSO regression were used to screen predictors using delayed onset of lactogenesis as the outcome variable. Risk prediction models were constructed based on three machine learning algorithms of Logistic regression, support vector, and random forest, respectively. The models were tuned by ten-fold cross-validation to filter out the best models.Results:Delayed onset of lactogenesis occurred in 140 of 338 prenatally overweight women, an incidence of 41.4%. Among the three predictive model performances, the random forest model had the highest area under the receiver operating characteristic curve, accuracy, precision, recall, and F1 value. The importance of each predictor was ranked according to the fandom forest algorithm, and in descending order of importance, they were breastfeeding 1 h after the birth of the newborn, number of previous deliveries, age, feeding mode 3 d postpartum, pregnancy complications, mode of delivery, number of breastfeeding 24 h postpartum, and monthly household income.Conclusions:Risk prediction models for delayed onset of lactogenesis in prenatally overweight women are constructed based on three machine learning algorithms, aiming to help provide a scientific basis for clinical healthcare professionals to take relevant decisions.

Objective To explore the impact of the sialic acid binding lectin-E(Siglec-E)on the inhibitory properties of parthenolide(PTL)against lipopolysaccharide(LPS)-induced M1 polarization of microglia(BV2).Methods ①Single cell sequencing data of Siglece related mouse brain tissue was obtained from Gene Expression Omnibus(GEO)database and divided into the WT group(n=3)and the Siglece-/-group(n=4).The microglia cells were screened,and the enrichment analysis was performed to analyze related differential genes and pathways.BV2 cells were constructed by the shRNA interference technique and were divided into NC-shRNA and Siglece-shRNA to detect the expression level of Siglec-E(Siglece).② NC-shRNA and Siglece-shRNA cells were respectively divided into the Control group,LPS group,PTL group and PTL+LPS group(n=3).The mRNA levels of markers of M1 polarization in microglia,iNOS,IL-1 β and IL-6,were detected by RT-qPCR.Siglecefl/fl and Cx3cr1cre mice were mated to obtain microglia-specific Siglece deletion(Siglecefl/fl×Cx3cr1cre)mice,and LPS-induced neuroinflammation model was established.③ Nine WT and Siglecefl/fl×Cx3cr1cre male mice were assigned to the Control group,LPS group and PTL+LPS group(n=3).RT-qPCR,immunofluorescence assay and Western blotting were used to verify the knock-out effect and polarization-related pathways,and to investigate the mechanism of Siglec-E affecting PTL inhibition of M1 polarization of microglia.Results Compared with the NC-shRNA group,the expression of Siglec-E in the Siglece-shRNA group was significantly decreased(P<0.01),indicating that the Siglec-E knock-down cell model was successfully established.With the stimulation of LPS,mRNA levels ofiNOS,IL-1 β and IL-6 were significantly up-regulated compared with the Control group both in shRNA cells and Siglece-shRNA cells(P<0.01).With the influence of PTL and LPS,the markers of M1 polarization in NC-shRNA cells mentioned before were significantly decreased(P<0.05),while for Siglice-shRNA cells,there were no significant changes in the markers of M1 polarization.PTL inhibited the phosphorylation of JNK and IκB protein(P<0.01)and the nuclear translocation of NF-κB in BV2 cells,down-regulated Siglec-E,and weakened the inhibitory effect.Compared with mice in the WT group,the expression of Siglec-E in microglia of Siglecefl/fl×Cx3cr1cre mice was decreased significantly(P<0.01),and the inhibitory effect of PTL on the phosphorylation of NF-κB in microglia of Siglecefl/fl×Cx3cr1cre mice was also decreased.Conclusion The absence of Siglec-E in microglia attenuates the inhibition of M1 polarization by the MAPK/NF-κB pathway targeted by PTL.