Breast cancer is a kind of malignant tumor with a complex mechanism， and its morbidity and mortality are increasing year by year， which seriously threatens women's health. At present， the main clinical treatments are surgical resection， radiotherapy， chemotherapy， and drug therapy， but they are often accompanied by side effects and adverse reactions， which affect the therapeutic effect. Traditional Chinese medicine （TCM） has the advantages of multi-component and multi-target treatment in the fight against breast cancer. The wnt/β-catenin signaling pathway is one of the classic pathways in cancer research. Abnormally activated Wnt/β-catenin signaling pathway inhibits β-catenin degradation by blocking the formation of Axin/glycogen synthase kinase 3β/adenomatous polyposis coli complex， thus promoting β-catenin nuclear metastasis， and it binds to T cell transcription factor/lymphoenhancer factor-1 to initiate downstream target genes and further interfere with the proliferation， migration， and invasion of tumor cells to affect the tumor process. Previous studies have shown that TCM monomers and compounds can mediate the Wnt/β-catenin signaling pathway to inhibit the malignant phenotype of breast cancer cells， thus playing an anti-breast cancer role， and the biochemical process involved in the regulation of therapeutic drugs has not been systematically combed. By analyzing and collating Chinese and foreign literature at the present stage， this paper discussed the association mechanism between Wnt/β-catenin signaling pathway and breast cancer and analyzed the internal mechanism of TCM monomers and compounds in mediating Wnt/β-catenin signaling pathway to exert anti-breast cancer effect. The statistical results showed that the flavonoids， alkaloids， and terpenoids in TCM monomers could target the Wnt/β-catenin signaling pathway and block the further development of malignant phenotype of breast cancer cells. TCM compounds with functions of clearing heat and detoxifying， promoting blood circulation and removing blood stasis， and tonifying kidney and liver were commonly used to intervene in the Wnt/β-catenin signaling pathway to prevent breast cancer. Compared with the current inhibitors of Wnt/β-catenin signaling pathway， the application of TCM monomers and compounds is expected to bring low-toxicity and high-efficiency breast cancer treatment drugs to the clinical practice， and the existing results provide a reference for the subsequent screening， research， and development of TCM small-molecule compounds and TCM compounds against breast cancer.

ObjectiveFor prepubertal and urgently treated malignant tumor patients, ovarian tissue cryopreservation and transplantation represent more appropriate fertility preservation methods. Current clinical practices often involve freezing ovarian tissue with high concentrations of cryoprotectants (CPAs) and thawing with water baths. These processes lead to varying degrees of toxicity and devitrification damage to ovarian tissue. Therefore, this paper proposes optimized methods for vitrification of ovarian tissues based on sodium alginate hydrogel encapsulation and magnetic induction nanowarming technology. MethodsFirstly, the study investigated the effects of sodium alginate concentration, the sequence of hydrogel encapsulation and CPAs loading on vitrification efficiency of encapsulated ovarian tissue. Additionally, the capability of sodium alginate hydrogel encapsulation to reduce the required concentration of CPAs was validated. Secondly, a platform combining water bath and magnetic induction nanowarming was established to rewarm ovarian tissue under various concentrations of magnetic nanoparticles and magnetic field strengths. The post-warming follicle survival rate, antioxidant capacity, and ovarian tissue integrity were evaluated to assess the efficacy of the method. ResultsThe study found that ovarian tissue encapsulated with 2% sodium alginate hydrogel exhibited the highest follicle survival rate after vitrification. The method of loading CPAs prior to encapsulation proved more suitable for ovarian tissue cryopreservation, effectively reducing the required concentration of CPAs by 50%. A combination of 8 g/L Fe₃O₄ nanoparticles and an alternating magnetic field of 300 Gs showed optimal warming effectiveness for ovarian tissue. Combining water bath rewarming with magnetic induction nanowarming yielded the highest follicle survival rate, enhanced antioxidant capacity, and preserved tissue morphology. ConclusionSodium alginate hydrogel encapsulation of ovarian tissue reduces the concentration of CPAs required during the freezing process. The combination of magnetic induction nanowarming with water bath provides an efficient method ovarian tissue rewarming. This study offers novel approaches to optimize ovarian tissues vitrification.

BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.

BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

The present study aimed to explore whether hydrogen sulfide (H₂S) improved hypoxic pulmonary hypertension (HPH) in rats by inhibiting aerobic glycolysis-pyroptosis. Male Sprague-Dawley (SD) rats were randomly divided into normal group, normal+NaHS group, hypoxia group, and hypoxia+NaHS group, with 6 rats in each group. The control group rats were placed in a normoxic (21% O₂) environment and received daily intraperitoneal injections of an equal volume of normal saline. The normal+NaHS group rats were placed in a normoxic environment and intraperitoneally injected with 14 μmol/kg NaHS daily. The hypoxia group rats were placed in a hypoxia chamber, and the oxygen controller inside the chamber maintained the oxygen concentration at 9% to 10% by controlling the N₂ flow rate. An equal volume of normal saline was injected intraperitoneally every day. The hypoxia+NaHS group rats were also placed in an hypoxia chamber and intraperitoneally injected with 14 μmol/kg NaHS daily. After the completion of the four-week modeling, the mean pulmonary artery pressure (mPAP) of each group was measured using right heart catheterization technique, and the right ventricular hypertrophy index (RVHI) was weighed and calculated. HE staining was used to observe pathological changes in lung tissue, Masson staining was used to observe fibrosis of lung tissue, and Western blot was used to detect protein expression levels of hexokinase 2 (HK2), pyruvate dehydrogenase (PDH), pyruvate kinase isozyme type M2 (PKM2), nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), GSDMD-N-terminal domain (GSDMD-N), Caspase-1, interleukin-1β (IL-1β) and IL-18 in lung tissue. ELISA was used to detect contents of IL-1β and IL-18 in lung tissue. The results showed that, compared with the normal control group, there were no significant changes in all indexes in the normal+NaHS group, while the hypoxia group exhibited significantly increased mPAP and RVHI, thickened pulmonary vascular wall, narrowed lumen, increased collagen fibers, up-regulated expression levels of aerobic glycolysis-related proteins (HK2 and PKM2), up-regulated expression levels of pyroptosis-related proteins (NLRP3, GSDMD-N, Caspase-1, IL-1β, and IL-18), and increased contents of IL-1β and IL-18. These changes of the above indexes in the hypoxia group were significantly reversed by NaHS. These results suggest that H₂S can improve rat HPH by inhibiting aerobic glycolysis-pyroptosis.
Animals ; Rats, Sprague-Dawley ; Male ; Hypertension, Pulmonary/metabolism* ; Glycolysis/drug effects* ; Hydrogen Sulfide/therapeutic use* ; Hypoxia/complications* ; Rats ; Pyroptosis/drug effects*

OBJECTIVE: To analyze the core acupoint selection rules and syndrome-based compatibility patterns of acupuncture for autism spectrum disorder (ASD) using data mining techniques. METHODS: Relevant literature of acupuncture for ASD was retrieved from CNKI, Wanfang, VIP, PubMed, and Web of Science. After applying inclusion and exclusion criteria, a prescription database was established based on the extracted effective data. Descriptive analysis was conducted on the frequency, meridian tropism, anatomical distribution, and specific point. High-frequency acupoints were visualized using Origin software. The Apriori algorithm in IBM SPSS Modeler 18.0 was used for association rule analysis of acupoint combinations. Cluster analysis of high-frequency acupoints was performed using IBM SPSS Statistics 26.0. The relationships between high-frequency syndromes and acupoints were visualized using Cytoscape 3.10.0. RESULTS: A total of 223 studies and 452 prescriptions were included, among which 223 were based on syndrome differentiation. A total of 205 acupoints were included with a cumulative frequency of 4 067. The top five most frequently used acupoints were Baihui (GV20), Sishenzhen, Zhisanzhen, Niesanzhen, and Neiguan (PC6). Acupoints were primarily from Jin's three-needle therapy, the governor vessel, scalp acupuncture, and the foot-taiyang bladder meridian, with a high proportion of acupoints located on the head and neck and the limbs. Among specific point, five-shu points, yuan-source points, and back-shu points were most frequently used. Association rule analysis revealed that the core acupoint group was Sishenzhen-Dingshenzhen-Zhisanzhen-Niesanzhen. Cluster analysis divided the top 20 high-frequency acupoints into four categories: governor vessel activation and brain awakening group, spleen strengthening and heart nourishing group, Jin's three-needle spirit-regulating group, and kidney-reinforcing and marrow-filling group. Clinically, the main syndrome patterns were kidney essence deficiency, hyperactivity of heart and liver fire, phlegm obstructing the heart orifices, dual deficiency of heart and spleen, and liver qi stagnation. CONCLUSION The core acupoint prescriptions of acupuncture for ASD are Sishenzhen, Dingshenzhen, Zhisanzhen, and Niesanzhen. The treatment emphasizes spirit regulation and mental tranquility, guided by the principles of harmonizing multiple zang-fu organs, regulating qi and blood, unblocking qi movement, and balancing yin and yang. Syndrome-based acupoint compatibility is recommended in clinical practice.
Humans ; Acupuncture Points ; Acupuncture Therapy ; Autism Spectrum Disorder/therapy* ; Data Mining ; Meridians

Ischemic stroke is the most common type of stroke,characterized by a complex pathophysiology in which inflammato-ry responses play a pivotal role,significantly affecting patient outcomes.Following a stroke,the disruption of the blood-brain barrier facilitates interactions between the central nervous system(CNS)and the peripheral immune system,triggering a cascade of inflammatory responses that exacerbate ischemic brain injury.Microglia(MG),as the primary immune cells of the CNS,are highly sensitive to cerebral ischemia and interact with various peripheral immune cells to regulate the inflammatory response.For instance,neutrophils participate in early MG-mediated in-flammatory responses through the release of neutrophil extracel-lular traps;T cells interact with MG,exerting bidirectional effects on the spread of brain inflammation and tissue repair.Additionally,other immune cells such as B cells,monocytes/macrophages,and dendritic cells contribute to these processes through complex immune regulatory networks,influencing MG's inflammatory functions and its capacity for tissue repair.This re-view delves into the interaction mechanisms between MG and pe-ripheral immune cells,providing novel insights into the dynamic changes of post-stroke inflammation and potential directions for precision therapies aimed at modulating immune balance.

Purpose To investigate the expression of SNRPA in gastric cancer and its effect on the proliferation,migration and invasion of gastric cancer cells.Methods The expression of SNRPA in gastric cancer tissues was ana-lyzed using The Cancer Genome Atlas(TCGA)database.The level of SNRPA was detected by immunohistochemistry(EnVision method),and its relationship with clinicopathological parameters was analyzed.Western blot was used to detect the expression of SNRPA in 15 cases of fresh gastric cancer and paracancerous tissues.The expression of SNRPA in MGC-803 and GES-1 cells was detected by immunofluorescence staining.The effects of SNRPA expression on the proliferation,migration and invasion of gastric cancer cell lines were determined by RNA interference technology and cell function assays,and the expression levels of Cyclin D1 protein and EMT-related proteins(E-cadherin,N-cadher-in)were detected by Western blot.Results The expression level of SNRPA in gastric cancer tissues was significantly higher than that in normal gastric mucosal tissues(P＜0.05),with an area under the curve(AUC)of 0.902 for diag-nostic accuracy.The Kaplan-Meier survival curves showed that the survival time of the group with high expression of SNRPA was shorter.SNRPA expression correlated with tumor size,Ki67,infiltration depth,and p53 status(P＜0.05).Compared with the corresponding control group,SNRPA silencing inhibited the proliferation,migration and in-vasion ability of gastric cancer cells,accompanied by decreased Cyclin D1 and N-cadherin expression and increased E-cadherin expression(P＜0.05).Conclusion SNRPA expression is upregulated in gastric cancer tissues and cell lines,promoting the proliferation,migration,and invasion of gastric cancer cells,and may be a potential molecular marker for gastric cancer.

Objective:To assess the effects of various leisure hobbies on the ability of daily living(ADL)and the quality of life among older adults in China.Methods:A cross-sectional study was conducted using data from the 4th Chinese Urban and Rural Elderly Living Conditions Sample Survey.We categorized community recreation hobbies into two types: dynamic hobbies, which include walking, playing ball games, dancing, and other physical activities, and static hobbies, which encompass activities that do not require physical exertion, such as reading, watching TV, and drawing.The EuroQol five-dimension scale(EQ-5D)-3L was employed to evaluate the quality of life.Older adults who maintained either dynamic or static hobbies for three consecutive years from 2017 to 2019 were initially screened.The relationship between different types of leisure hobbies and ADL scores, as well as quality of life at various time points, was analyzed using repeated-measures ANOVA.Subsequently, propensity scores were matched based on gender, age, ethnicity, marital status, literacy, and chronic diseases, and five machine learning algorithms were utilized to identify the optimal model for predicting fully independent ADL and EQ-5D effect values.Based on the optimal predictive model, we conducted Shapley Additive Explanations(SHAP)and global analysis.Results:The total number of older adults who participated in the survey over the three consecutive years from 2017 to 2019 was 4 912.Among these, 2 594 individuals maintained the same hobby habits throughout this period, comprising 1 362 males and 1 232 females, with an average age of(70.81±6.71)years.Statistically significant differences in ADL scores and EQ-5D scores were observed between the dynamic and static leisure hobby groups(all P<0.05)across various time points, hobby group classifications, and interaction levels.Older adults engaged in dynamic hobbies exhibited superior ADL functioning and higher EQ-5D scores in comparison to those in the static hobby group, with these differences remaining statistically significant following propensity score matching(all P<0.05).In predicting full ADL independence, the SHAP value for dynamic hobbies ranked second, while for predicting an EQ-5D weighted value of 1, the SHAP value for dynamic hobbies ranked ninth.Overall, the type of dynamic hobby demonstrated a positive trend in its effect on the ability to perform daily activities independently and in achieving an EQ-5D weighted value of 1. Conclusions:In comparison to static hobbies, Chinese older adults who engage in dynamic hobbies exhibit improved daily mobility and a higher quality of life.Furthermore, dynamic hobbies positively influence the capacity for full independence in daily activities, as evidenced by an EQ-5D weighted value of 1.

Objective:To construct a nanoparticle vaccine displaying the prefusion F (preF) protein of respiratory syncytial virus (RSV) using the I53-50 protein nanoparticle platform, and to systematically evaluate its immunogenicity and protective efficacy.Methods:The RSV preF trimer antigen was genetically fused to I53-50A and assembled in vitro with I53-50B to form preF-I53-50 nanoparticles, theoretically displaying 20 preF antigens per particle. The structure and purity were characterized by size-exclusion chromatography, SDS-PAGE, and negative-stain electron microscopy. BALB/c mice were intramuscularly immunized with varying doses (1 μg or 5 μg) of preF antigen or an equimolar amount of preF-I53-50 nanoparticles. Humoral immunity, B-cell responses, and protective efficacy were assessed following intranasal viral challenge.Results:The preF-I53-50 nanoparticles self-assembled into spherical structures (50-60 nm in diameter) with uniformly arrayed antigens. The nanoparticle vaccine enhanced RSV-specific IgG1 and IgG2a antibody responses, promoting a Th1-biased immune profile. At equimolar preF doses, the neutralizing antibody titers induced by 1 μg and 5 μg nanoparticle formulations were 2.8-fold and 2.3-fold higher, respectively, than those elicited by preF alone ( P<0.05). Notably, even the low-dose nanoparticle group outperformed the high-dose preF group (1.6-fold increase). Viral challenge experiments demonstrated that preF-I53-50 effectively suppressed pulmonary viral replication, mitigated pathological damage, and induced stronger germinal center and memory B-cell responses, suggesting enhanced B-cell affinity maturation and long-term immune memory. Conclusion:The preF-I53-50 vaccine improves the immunogenicity and protective efficacy of RSV preF through multivalent antigen display.