ObjectiveTo elucidate the expression patterns of CYHR1 in colorectal cancer （CRC） and its regulatory role in tumour cell proliferation and metastasis-related phenotypes. MethodsExpression differences of CYHR1 between colorectal cancer tissues and adjacent normal tissues were analyzed using data from The Cancer Genome Atlas （TCGA） database， and the association between CYHR1 expression and patient survival was evaluated. A CRC cell model with CYHR1 overexpression was established， and changes in the expression of related molecules were verified by quantitative real-time PCR （qRT-PCR）. Cell proliferation and metastatic potential were assessed using MTT assays， colony formation assays， and Transwell migration and invasion assays to evaluate the effects of CYHR1 and MALAT1 on CRC cell proliferation and migration/invasion， including combined intervention experiments. ResultsDatabase analysis revealed that CYHR1 is significantly overexpressed in CRC tissues， and that this overexpression is significantly associated with poorer overall survival. Compared with normal control cells， mRNA expression levels of CYHR1 were markedly elevated in CRC cell lines （HCT116， RKO， etc.） （P=0.002； P=0.006）. Functional experiments demonstrated that CYHR1 overexpression significantly promoted the proliferation （P＜0.001）， colony formation （P＜0.001）， migration （P=0.002） and invasive capacity （P=0.002） of HCT116 cells； whereas knockdown of either CYHR1 or MALAT1 inhibited CRC cell proliferation and reduced their migration （P=0.006） and invasive capacity （P=0.006）. In the context of CYHR1 overexpression， combined knockdown of MALAT1 partially reversed the promotional effect of CYHR1 on CRC cell proliferation， migration and invasion （P＜0.001； P=0.001）. Mechanistic studies further revealed that CYHR1 overexpression upregulates MALAT1 mRNA expression levels （P＜0.001）， whilst CYHR1 knockdown leads to downregulation of MALAT1 expression （P＜0.001）； MALAT1 knockdown likewise downregulates CYHR1 expression， suggesting a mutually reinforcing positive regulatory relationship between the two. Furthermore， CYHR1 can activate the IL-6/STAT3 signalling axis， whilst knockdown of IL-6 or STAT3 inhibited the upregulation of MALAT1 expression by CYHR1 （P=0.031； P=0.006）. ConclusionCYHR1 is abnormally overexpressed in colorectal cancer and is closely associated with poor prognosis. CYHR1 may promote the proliferation， migration， and invasion of colorectal cancer （CRC） cells by activating the IL-6/STAT3/MALAT1 axis.

Objective To explore the effect and mechanism of Gegen Qinlian decoction (GQD) on myocardial injury induced by high-fat diet combined with streptozotocin in type 2 diabetic mice through network pharmacology and animal experiments. MethodsNetwork pharmacology was applied to screen the potential signaling pathways of GQD in the intervention of diabetic heart disease (DHD). A total of 36 specific pathogen-free (SPF) male C57BL/6J mice aged 8 weeks were acclimatized for 1 week and randomly divided into a normal control group (n=6) and a model group (n=30). Mice in the model group were fed a 60% high-fat diet and 5% glucose water for 4 weeks, and were intraperitoneally injected with 100 mg/kg streptozotocin after 12 h of fasting to establish the type 2 diabetes model. Mice in the normal control group were given a conventional diet and drinking water, and were intraperitoneally injected with an equal volume of citrate buffer after 12 h of fasting at the same time point. After successful establishment of the diabetic mouse model, the model mice were randomly divided into five groups according to body weight: model control group, GQD low-dose group, GQD medium-dose group, GQD high-dose group, and metformin positive control group, with 6 mice in each group. After 6 weeks of intragastric administration of corresponding doses of GQD, metformin, and ddH₂O respectively, all mice in the six groups (one normal control group and five model groups) were anesthetized with inhaled 2% isoflurane, blood samples were collected via eyeball enucleation, and heart tissues were harvested after euthanasia by cardiac exsanguination. Hematoxylin-eosin (HE) staining and wheat germ agglutinin (WGA) staining were performed to observe the pathological damage of myocardial tissues and measure the cross-sectional area of cardiomyocytes. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of pro-inflammatory cytokines in serum. Immunohistochemical staining was adopted to determine the protein expression levels of receptor for advanced glycation end products (RAGE), nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3), and phosphorylated nuclear factor-κB p65 (p-p65) in myocardial tissues. ResultsNetwork pharmacology analysis revealed that the targets of GQD against DHD were significantly enriched in inflammatory pathways, including the advanced glycation end products (AGEs)-RAGE axis, tumor necrosis factor (TNF) and interleukin-17 (IL-17) signaling pathways. HE and WGA staining results showed that, compared with the normal control group, the model control group exhibited myocardial fiber rupture, cardiomyocyte swelling, and significantly increased cardiomyocyte cross-sectional area (P<0.05). Compared with the model control group, GQD intervention significantly ameliorated the pathological changes, including myocardial fiber rupture, cell swelling, and cardiomyocyte hypertrophy. The results of immunohistochemical staining and ELISA indicated that compared with the model control group, GQD intervention markedly downregulated the protein expression of RAGE, NLRP3, and p-p65 in myocardial tissues, as well as the levels of serum pro-inflammatory cytokines (P<0.05). ConclusionGQD can effectively alleviate myocardial pathological injury in diet-induced type 2 diabetic mice, and its mechanism may be related to inhibiting the activation of the RAGE/p-p65/NLRP3 signaling pathway and reducing myocardial inflammatory response.

Objective:To develop and validate a predictive model for massive hemorrhage during brain tumor resection in pediatric patients.Methods:A retrospective analysis was performed on the clinical data from pediatric patients who underwent elective brain tumor resection under general anesthesia at the Women and Children′s Medical Center Affiliated to Guangzhou Medical University from December 2016 to October 2023. The patients were randomly divided into model group and internal validation group in a ratio of 8∶2. Pediatric patients who underwent elective brain tumor resection under general anesthesia at Qilu Hospital of Shandong University from January 2021 to July 2024 were selected and served as external validation group. Relevant characteristic variables were screened through Lasso regression. A multivariate logistic regression was used to develop the model and plot the nomogram for intraoperative massive hemorrhage. The performance of the model was evaluated using the area under the receiver operating characteristic curve and calibration curve.Results:Through Lasso regression and multivariate logistic regression analyses, 11 independent influencing factors were identified: age ( OR=0.323, 95% confidence interval [ CI]: 0.280-0.374, P<0.001), weight ( OR=0.164, 95% CI: 0.135-0.199, P<0.001), activated partial thromboplastin time ( OR=1.133, 95% CI: 1.036-1.239, P=0.006), thrombin time ( OR=1.141, 95% CI: 1.048-1.243, P=0.002), red blood cell count ( OR=0.941, 95% CI: 0.888-0.996, P=0.035), hemoglobin concentration ( OR=0.873, 95% CI: 0.822-0.926, P<0.001), platelet count ( OR=1.062, 95% CI: 1.001-1.127, P=0.048), maximum tumor diameter ( OR=2.384, 95% CI: 2.241-2.536, P<0.001), tumor invasiveness ( OR=2.376, 95% CI: 2.071-2.726, P<0.001), hydrocephalus ( OR=2.409, 95% CI: 2.139-2.713, P<0.001), and centered midline structure ( OR=0.509, 95% CI: 0.465-0.557, P<0.001). Based on this, a nomogram prediction model was established. The receiver operating characteristic curve showed that the area under the curve of this model in predicting the risk of massive hemorrhage during brain tumor resection was 0.936 (95% CI: 0.90-0.959) in model group, 0.863 (95% CI: 0.744-0.948) in internal validation group, and 0.855 (95% CI: 0.726-0.955) in external validation group. The calibration curve indicated good model consistency, and the Hosmer-Lemeshow goodness-of-fit test result showed a P value of 0.979 ( P>0.05). Conclusions:Age, body weight, activated partial thromboplastin time, thrombin time, red blood cell count, hemoglobin concentration, platelet count, maximum tumor diameter, tumor invasiveness, hydrocephalus and midline structure are independent influencing factors for major bleeding during brain tumor resection in pediatric patients, and the prediction model established based on this histogram has high accuracy.

Objective:To explore the association between the systemic immune-inflammation index (SII) and obesity metabolic phenotypes, as well as metabolic features in children and adolescents.Methods:A cross-sectional study was conducted using the random cluster sampling method from March 2023 to May 2024. Children and adolescents aged 9-17 years in Guangzhou were surveyed through questionnaires, physical measurements, and blood tests. According to BMI and metabolic status, participants were classified into normal-weight groups [metabolically healthy normal weight (MHNW) and metabolically unhealthy normal weight (MUNW)] and overweight/obese groups [metabolically healthy overweight/obese (MHO/O) and metabolically unhealthy overweight/obese (MUO/O)]. After natural log-transformation of SII values (lnSII), multinomial logistic regression was used to assess the association between SII and obesity metabolic phenotypes, while binary logistic regression was applied to assess the relationship between SII and metabolic phenotypes in the overweight/obese subgroup. Linear regression model and restricted cubic spline (RCS) were employed to examine the relationship between SII and metabolic features among the entire population.Results:A total of 3 749 participants were included. After adjusting for covariates, for every unit increase in lnSII, the risk of MHO/O and MUO/O increased by 93% ( OR=1.93, 95% CI: 1.56-2.40, P<0.001) and 156% ( OR=2.56, 95% CI: 2.02-3.25, P<0.001), respectively. In the overweight/obesity subgroup, for every unit increase in lnSII, the risk of MUO/O increased by 37% ( OR=1.37, 95% CI: 1.01-1.87, P=0.045). Linear regression model and RCS showed that lnSII was positively correlated with systolic blood pressure (SBP) and diastolic blood pressure (DBP) (SBP: β=1.39, 95% CI: 0.67-2.11, P<0.001; DBP: β=1.27, 95% CI: 0.79-1.75, P<0.001). lnSII also had a non-linear relationship with triglyceride ( Pnonlinear=0.032) and high-density lipoprotein cholesterol ( Pnonlinear=0.002). Conclusion:Elevated SII levels are associated with unfavorable obesity metabolic phenotypes, higher blood pressure, and altered lipid profiles in children and adolescents. SII may be a potential driving factor for metabolic heterogeneity in children and adolescents.

Objective To construct and implement a swallowing disorder assessment and management program for tracheal intubated patients after extubation based on the 4R crisis management theory,providing standardized and scientific interventions for oral feeding.Methods Utilizing the expert meeting method with the 4R crisis management theory framework,a swallowing disorder assessment and management program was developed for post-extubation tracheal intubated patients.A convenience sampling method was employed to select patients with tracheal intubations treated from July to December 2023 in the emergency ICU,central ICU,and cardiovascular surgery ICU of a tertiary hospital in Zhejiang Province.The patients treated from October to December were assigned to an experimental group(n=68),while those treated from July to September were designated as a control group(n=58).The experimental group received the 4R crisis management-based intervention,whereas the control group received standard ICU assessment and management.Outcomes indicators included the incidence of post-extubation swallowing disorders,time to first oral intake,incidence of aspiration during initial feeding,nasogastric and nasointestinal tube placement duration,incidence of aspiration pneumonia during hospitalization,re-intubation rates,ICU readmission rates,ICU stay duration,and total hospitalization days.Results Of the initially recruited subjects,68 in the experimental group and 54 in the control group were included in the final analysis.After the intervention,the experimental group exhibited significantly lower rates of post-extubation swallowing disorders,shorter time to first liquid oral intake,aspiration incidence during first feeding,shorter durations of nasogastric and nasointestinal tube placement,aspiration pneumonia,ICU readmission compared to the control group(P＜0.05).No significant differences were observed between the groups in time to first regular oral intake,re-intubation rates(P＞0.05).Conclusion The risk management program for dysphagia following tracheal extubation based on the 4R crisis management theory is scientifically robust and safe,offering a valuable reference for clinical assessments and management of swallowing and eating post-extubation in tracheal intubated patients.

Objective To explore the repair effect and mechanism of human amniotic mesenchymal stem cells(hAMSCs)on endometrial injury.Methods hAMSCs were isolated using a two-enzyme digestion and then cultured.The third-passage(P3)cells were harvested to detect the surface markers by flow cytometry and to identify their trilineage differentiation potentials.Eighteen nulliparous female SD rats(8～9 weeks old,weighing 250～280 g)were randomly divided into 3 groups(n=6):normal control group,model group,and hAMSCs group.A rat model of intrauterine adhesions(IUA)was established in SD rats by using curettage combined with lipopolysaccharide(LPS)infection.In 2 weeks after modeling,the hAMSCs group received a bilateral uterine horn transplantation of 0.2 mL hAMSCs(1.0×10? cells/mL),while the model group received a same volume of PBS into both uterine horns.All rats were sacrificed in 2 weeks after transplantation.HE and Masson staining was used to observe endometrial thickness and gland number as well as endometrial fibrosis area.RT-qPCR and Western blotting were performed to detect the mRNA and protein levels of TGF-β1,Smad3,Smad7,epithelial-mesenchymal transition(EMT)markers(E-cadherin,Vimentin),fibrosis factor α-SMA,and endometrial estrogen receptor(ER)and progesterone receptor(PR)in endometrial tissues.Results The obtained cells were identified as hAMSCs due to the characteristics of surface markers and differentiation potentials.Compared with the normal control group,the model group showed decreased endometrial thickness,reduced gland number,increased fibrosis area,and enhanced mRNA and protein levels of fibrosis-related factors TGF-β1,Smad3,Vimentin,and α-SMA(P<0.01),while down-regulation of fibrosis-inhibiting molecule Smad7,the EMT marker E-cadherin,and endometrial receptors ER and PR at both mRNA and protein levels(P<0.01).hAMSCs transplantation increased endometrial thickness and gland number,decreased fibrosis area,and down-regulated mRNA expression of the aforementioned fibrosis-related factors(P<0.01),and up-regulated the mRNA expression levels of Smad7,E-cadherin,ER,and PR(P<0.01).The hAMSCs group also exhibited obviously down-regulated protein levels of TGF-β1,Smad3,and α-SMA(P<0.05),while enhanced protein levels of Smad7 and PR(P<0.05).Conclusion Intrauterine transplantation of hAMSCs can promote the repair of endometrial injury,and inhibits endometrial EMT and fibrosis through the TGF-β1/Smad7 signaling pathway.

ObjectiveTo explore the role of lead exposure in the phenotypic transformation of vascular smooth muscle cells (VSMC), and to provide new insights for the mechanism of lead impact on vascular lesions. MethodsMouse aortic smooth muscle cells (MOVAS) were divided into a control group (0 μmol·L^-1), low concentration lead groups (0.1, 1, 5, and 10 μmol·L^-1), and high concentration lead groups (15, 25, and50 μmol·L^-1). MTT assays were used to assess the proliferation of the cells, and scratch assays were implicated to measure migration ability of the cells. Fluorescence quantitative PCR was employed to determine levels of mRNA expression for smooth muscle actin α (α⁃SMA), smooth muscle 22 alpha (SM22α), synthetic phenotype-related genes osteopontin (OPN), matrix metalloproteinase 9 (MMP9), and the transcription factor SOX9. Immunoblotting was used to determine levels of protein expression for α-SMA, OPN, and MMP9. ResultsProliferation of MOVAS was observed under the lead ions concentrations of 0‒50 µmol·L^-1, with a significant increase of proliferation compared to the control group at the concentrations of 5‒50 µmol·L^-1 (all P<0.05). The migration ability of cells gradually increased at the concentrations of 0‒10 µmol·L^-1, with a significant increase at 5 (q=4.574, P=0.003) and 10 µmol·L^-1 (q=10.570, P<0.001) compared to the control group. The 10 µmol·L^-1 lead ions significantly reduced the levels of mRNA expression for vascular smooth muscle contractile phenotype genes α⁃SMA (q=7.426, P<0.001) and SM22α (q=4.766, P=0.001), while significantly increasing the levels of mRNA expression for OPN (q=11.330, P<0.001), MMP9 (q=7.842, P<0.001), and SOX9 (q=11.120, P<0.001) genes. Furthermore, the 10 µmol·L^-1 lead ions significantly reduced the levels of protein expression for the vascular smooth muscle contractile phenotype marker α-SMA protein (q=2.897, P=0.049), while significantly increasing the levels of protein expression for the synthetic markers OPN (q=3.188, P=0.031) and MMP9 (q=3.292, P=0.026), compared to the control group. ConclusionTreatment with lead in vitro induced VSMC to differentiate from contractile phenotype to synthetic phenotype, indicating that a certain dose of lead exposure might be detrimental to the cardiovascular system.

Alzheimer's disease (AD) is a neurodegenerative disease with clinical hallmarks of progressive cognitive impairment. Synergistic effects of the Aβ-Tau cascade reaction are tightly implicated in AD pathology, and microglial NLRP3 inflammasome activation drives neuronal tauopathy. However, the underlying mechanism of how Aβ mediates NLRP3 inflammasome remains unclear. Herein, we determined that oligomeric Aβ (o-Aβ) bound to microglial Kv2.1 and promoted Kv2.1-dependent potassium efflux to activate NLRP3 inflammasome resulting in neuronal tauopathy by using Kv2.1 inhibitor drofenine (Dfe) as a probe. The underlying mechanism has been intensively investigated by assays with Kv2.1 knockdown in vitro (si-Kv2.1) and in vivo (AAV-ePHP-si-Kv2.1). Dfe deprived o-Aβ of its capability to promote microglial NLRP3 inflammasome activation and neuronal Tau hyperphosphorylation by inhibiting the Kv2.1/JNK/NF-κB pathway while improving the cognitive impairment of 5×FAD-AD model mice. Our results have highly addressed that the Kv2.1 channel is required for o-Aβ-driven microglial NLRP3 inflammasome activation and neuronal tauopathy in AD model mice and highlighted that Dfe as a Kv2.1 inhibitor shows potential in the treatment of AD.

Dendritic cells (DCs) serve as the primary antigen-presenting cells in autoimmune diseases, like rheumatoid arthritis (RA), and exhibit distinct signaling profiles due to antigenic diversity. Type II collagen (CII) has been recognized as an RA-specific antigen; however, little is known about CII-stimulated DCs, limiting the development of RA-specific therapeutic interventions. In this study, we show that CII-stimulated DCs display a preferential gene expression profile associated with migration, offering a new perspective for targeting DC migration in RA treatment. Then, saikosaponin D (SSD) was identified as a compound capable of blocking CII-induced DC migration and effectively ameliorating arthritis. Optineurin (OPTN) is further revealed as a potential SSD target, with Optn deletion impairing CII-pulsed DC migration without affecting maturation. Function analyses uncover that OPTN prevents the proteasomal transport and ubiquitin-dependent degradation of C-C chemokine receptor 7 (CCR7), a pivotal chemokine receptor in DC migration. Optn-deficient DCs exhibit reduced CCR7 expression, leading to slower migration in CII-surrounded environment, thus alleviating arthritis progression. Our findings underscore the significance of antigen-specific DC activation in RA and suggest OPTN is a crucial regulator of CII-specific DC migration. OPTN emerges as a promising drug target for RA, potentially offering significant value for the therapeutic management of RA.

Acute kidney injury (AKI) has high morbidity and mortality, but effective clinical drugs and management are lacking. Previous studies have suggested that macrophages play a crucial role in the inflammatory response to AKI and may serve as potential therapeutic targets. Emerging evidence has highlighted the importance of forkhead box protein O1 (FoxO1) in mediating macrophage activation and polarization in various diseases, but the specific mechanisms by which FoxO1 regulates macrophages during AKI remain unclear. The present study aimed to investigate the role of FoxO1 in macrophages in the pathogenesis of AKI. We observed a significant upregulation of FoxO1 in kidney macrophages following ischemia-reperfusion (I/R) injury. Additionally, our findings demonstrated that the administration of FoxO1 inhibitor AS1842856-encapsulated liposome (AS-Lipo), mainly acting on macrophages, effectively mitigated renal injury induced by I/R injury in mice. By generating myeloid-specific FoxO1-knockout mice, we further observed that the deficiency of FoxO1 in myeloid cells protected against I/R injury-induced AKI. Furthermore, our study provided evidence of FoxO1's pivotal role in macrophage chemotaxis, inflammation, and migration. Moreover, the impact of FoxO1 on the regulation of macrophage migration was mediated through RhoA guanine nucleotide exchange factor 1 (ARHGEF1), indicating that ARHGEF1 may serve as a potential intermediary between FoxO1 and the activity of the RhoA pathway. Consequently, our findings propose that FoxO1 plays a crucial role as a mediator and biomarker in the context of AKI. Targeting macrophage FoxO1 pharmacologically could potentially offer a promising therapeutic approach for AKI.