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.

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.

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.

Objective: To investigate the differences in three-dimensional maxillary and palatal parameters as well as midpalatal suture maturation stages between Skeletal Class I malocclusion patients with maxillary transverse deficiency (MTD) and normal maxillary transverse development, in order to provide clinical guidance for orthodontic diagnosis and treatment Methods: This study was approved by the institutional ethics committee and informed consent was obtained. Cone-bean CT data from 66 Skeletal Class I malocclusion patients [22 males, 44 females; age (14.39 ± 1.68) years] were analyzed. Based on Yonsei University width analysis, participants were divided into two groups: the maxillary transverse deficiency group [n = 33, age (14.34 ± 1.99) years] and the maxillary transverse normal group [n = 33, age (14.43 ± 1.33) years]. Parameters compared included midpalatal suture maturation stages (A-E), maxillary skeletal width on nasal floor (NF) and maxillary skeletal width on hard palate (HP), maxillary arch width on buccal alveolar crest (BAC) and maxillary arch width on lingual alveolar crest (LAC), palatal vault height, palatal bone thickness, palatal length, surface area, and volume Results: The proportion of patients with palatal suture maturity at stages D+E in the maxillary transverse deficiency group (33%, 11/33) was lower than that in the normal maxillary transverse development group (45%, 15/33), showing no statistically significant difference between the two groups. Compared to the maxillary transverse normal group, significant reductions were observed in the maxillary transverse deficiency group for maxillary widths [NF: (67.63 ± 3.74) mm vs. (71.49 ± 5.11) mm; HP: (64.60 ± 3.53) mm vs. (68.40 ± 4.64) mm], dental arch widths [BAC: (56.88 ± 2.25) mm vs. (59.81 ± 2.71) mm; LAC: (33.90 ± 1.89) mm vs. (36.91 ± 2.20) mm], and palatal surface area [(1 170.80 ± 126.48) mm2 vs. (1 264.76 ± 140.10) mm2]. No significant differences were noted in palatal height, bone thickness, length, or volume. Conclusion Skeletal Class I malocclusion patients with MTD have narrowed maxillary and dental arch widths, reduced palatal surface area, and delayed midpalatal suture maturation. Early maxillary expansion is recommended to harmonize jaw relationship.

Traumatic main bronchus rupture is a relatively rare injury in thoracic trauma, which is extremely critical, with a mortality rate as high as 70% - 80%. The complete rupture and displacement of the traumatic cervical trachea can lead to asphyxia, hypoxia, and cardiac arrest, even death of the patient in a short time. We performed emergency surgery with the support of extracorporeal membrane oxygenation for a case of traumatic cervical tracheal trunk complete rupture and displacement combined with cardiac arrest and achieved a successful rescue. We summarized our experience and found that timely surgical reconstruction of the airway is the key to increasing the traumatic main bronchus rupture survival of patients.
Humans ; Extracorporeal Membrane Oxygenation ; Heart Arrest/etiology* ; Rupture ; Trachea/surgery*

Doxorubicin (Dox) is an anthracycline drug widely applied in various malignancies. However, the fatal cardiotoxicity induced by Dox limits its clinical application. Post-transcriptional protein modification via ubiquitination/deubiquitination in cardiomyocytes mediates the pathophysiological process in Dox-induced cardiotoxicity (DIC). In this study, we aimed to clarify the regulatory role and mechanism of a deubiquitinating enzyme, ubiquitin-specific peptidase 13 (USP13), in DIC. RNA-seq analysis and experimental examinations identified that cardiomyocyte-derived USP13 positively correlated with DIC. Mice with cardiac-specific deletion of USP13 were subjected to Dox modeling. Adeno-associated virus serotype 9 (AAV9) carrying cTNT promoter was constructed to overexpress USP13 in mouse heart tissues. Cardiomyocyte-specific knockout of USP13 exacerbated DIC, while its overexpression mitigated DIC in mice. Mechanistically, USP13 deubiquitinates the stimulator of interferon genes (STING) and promotes the autolysosome-related degradation of STING, subsequently alleviating cardiomyocyte inflammation and death. Our study suggests that USP13 serves a cardioprotective role in DIC and indicates USP13 as a potential therapeutic target for DIC treatment.

Food-derived bioactive peptides (FBPs), particularly those with ten or fewer amino acid residues and a molecular weight below 1300 Da, have gained increasing attention for their safe, diverse structures and specific biological activities. The development of FBP-based functional foods and potential medications depends on understanding their structure‒activity relationships (SARs), stability, and bioavailability properties. In this review, we provide an in-depth overview of the roles of FBPs in treating various diseases, including Alzheimer's disease, hypertension, type 2 diabetes mellitus, liver diseases, and inflammatory bowel diseases, based on the literature from July 2017 to Mar. 2023. Subsequently, attention is directed toward elucidating the associations between the bioactivities and structural characteristics (e.g., molecular weight and the presence of specific amino acids within sequences and compositions) of FBPs. We also discuss in silico approaches for FBP screening and their limitations. Finally, we summarize recent advancements in formulation techniques to improve the bioavailability of FBPs in the food industry, thereby contributing to healthcare applications.
Humans ; Peptides/therapeutic use* ; Structure-Activity Relationship ; Functional Food ; Diabetes Mellitus, Type 2/drug therapy* ; Biological Availability ; Alzheimer Disease/drug therapy* ; Inflammatory Bowel Diseases/drug therapy* ; Hypertension/drug therapy* ; Liver Diseases/drug therapy* ; Bioactive Peptides, Dietary

Cancer is a highly deadly disease, with breast cancer, cervical cancer, endometrial cancer, and ovarian cancer being the most prevalent in women. However, traditional cancer treatments present challenges due to their strong toxic side effects and adverse reactions. Numerous studies have demonstrated that natural products derived from various plants possess therapeutic and preventive properties against cancer. These phytochemicals have been extensively investigated as a potential alternative to conventional chemotherapy drugs, owing to their safety and efficacy. This article provides a comprehensive review of the recent advances in the chemoprevention and mechanisms of phytochemicals against the four major female cancers. The focus will be on how these phytochemicals regulate cancer cell proliferation, apoptosis, invasion, and metastasis to impede cancer progression. Given their extensive clinical applications, phytochemicals hold great promise in the field of cancer treatment. It hopes that this review will inspire more researchers to explore the potential of these natural compounds in combating female cancers.

Human naïve pluripotent stem cells (PSCs) hold great promise for embryonic development studies. Existing induction and culture strategies for these cells, heavily dependent on MEK inhibitors, lead to widespread DNA hypomethylation, aberrant imprinting loss, and genomic instability during extended culture. Here, employing high-content analysis alongside a bifluorescence reporter system indicative of human naïve pluripotency, we screened over 1,600 chemicals and identified seven promising candidates. From these, we developed four optimized media-LAY, LADY, LUDY, and LKPY-that effectively induce and sustain PSCs in the naïve state. Notably, cells reset or cultured in these media, especially in the LAY system, demonstrate improved genome-wide DNA methylation status closely resembling that of pre-implantation counterparts, with partially restored imprinting and significantly enhanced genomic stability. Overall, our study contributes advancements to naïve pluripotency induction and long-term maintenance, providing insights for further applications of naïve PSCs.
Humans ; DNA Methylation/drug effects* ; Genomic Instability ; Pluripotent Stem Cells/metabolism* ; Cell Culture Techniques/methods* ; Cells, Cultured

Objective To investigate the potential molecular mechanism of modified Shaofu Zhuyu Decoction in treating endometriosis from the perspective of autophagy in ectopic endometrial tissue cells mediated by the epidermal growth factor receptor (EGFR)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. Methods Seventy-two female SD rats were divided into the blank group,the model group,the gestrinone group and the modified Shaofu Zhuyu Decoction high-,medium-,and low-dose groups according to the random number table method,12 rats in each group. Construction of an endometriosis model used auto-transplantation method. The rats in the gestrinone group were gavaged with 0.25mg/(kg·d) of gestrinone suspension,the rats in the modified Shaofu Zhuyu Decoction high-,medium-,and low-dose groups were gavaged with 30,15,and 7.5g/(kg·d) of modified Shaofu Zhuyu Decoction,respectively,and the rats in the blank and model groups were gavaged with an equal amount of physiological saline,respectively. 4 weeks of continuous treatment. At the end of drug administration,2 U of oxytocin was given to each rat to induce contractions,and the pain of the rats was observed;the weight and volume of ectopic endometrial tissue of the rats were recorded. Hematoxylin-eosin (HE) staining method was used to observe the pathological changes of rat endometrial tissue in each group;transmission electron microscope was used to observe the ultrastructure of rat endometrial tissue. Enzyme-linked immunosorbent assay was used to detect the serum levels of oestradiol (E2),progesterone (P),interleukin-1β (IL-β),interleukin-6 (IL-6),tumour necrosis factor-α (TNF-α),transforming growth factor-β (TGF-β),epidermal growth factor (EGF),and EGFR in the rat serum of each group. Mean fluorescence intensity of autophagy-associated protein yeast Atg6 homologue (Beclin-1) and microtubule-associated protein 1 light chain 3 (LC3B) in endometrial tissues of rat endometrium in each group by immunofluorescence assay. Protein expression levels of EGFR,PI3K,phosphorylated phosphatidylinositol 3-kinase (p-PI3K),AKT,and phosphorylated protein kinase B (p-AKT) in endometrial tissues of rats in each group were examined by protein blotting. Real-time fluorescence PCR was used to detect EGFR,PI3K,and AKT mRNA expression levels.Results Compared with the blank group,the rats in the model group showed typical torsion reaction,ectopic endometrial tissue was visible in the abdominal wall,and the HE result showed thickening of the ectopic endometrial tissue,mesenchymal hyperplasia,and glandular dilatation;only autophagic vesicles were seen in the ultrastructure,and no obvious autophagic structures were seen in some of the fields of view;Serum E2,IL-6,IL-1β,TNF-α,TGF-β,EGF,and EGFR levels were elevated and P levels were decreased in rats;autophagy-related proteins Beclin-1 and LC3B decreased in average fluorescence intensity;protein expressions of EGFR,PI3K,AKT,p-PI3K,p-AKT was elevated;and mRNA expressions of EGFR,PI3K,AKT was elevated (P<0.05). Compared with the model group,the gestrinone group and modified Shaofu Zhuyu Decoction high-,medium-dose groups showed a decrease in the number of torsion and a prolongation of the latency time of the torsion response,a decrease in the volume and mass of ectopic endometrial tissues,a decrease in the degree of pathological damage as shown by HE,and the appearance of lysosomes,autophagolysosomes and autophagic vesicles in different degrees in the ultrastructure;Serum E2,IL-6,IL-1β,TNF-α,TGF-β,EGF,EGFR levels were decreased,and P levels were increased;the mean fluorescence intensity of autophagy-related proteins Beclin-1 and LC3B was increased;the protein expression of EGFR,PI3K,AKT,p-PI3K,and p-AKT was decreased;and the mRNA expression of EGFR,PI3K,and AKT was decreased (P<0.05). Conclusion The mechanism of modified modified Shaofu Zhuyu Decoction for the treatment of endometriosis may be related to the induction of autophagy of ectopic endometrial tissues mediated by the EGFR/PI3K/AKT signaling pathway and the attenuation of inflammatory responses.