Abstract:
Objective　To investigate the effects of Trpc6 knockout on chronic lipopolysaccharide （LPS）-induced myocardial inflammation and fibrosis in mice and its potential mechanisms.Methods　Male C57BL/6 wild-type （WT） mice and Trpc6 knockout （Trpc6-/-） mice of the same background were randomly divided into four groups： WT control， WT+LPS （200 μg/kg）， Trpc6-/- control， and Trpc6-/-+LPS （200 μg/kg）. Group with LPS received intraperitoneal LPS injections for 21 consecutive days to induce chronic myocardial inflammatory injury. Cardiac ultrasound assessed changes in left ventricular ejection fraction （EF）， left ventricular shortening fraction （FS）， and cardiac output （CO）. Hematoxylin and eosin （HE） staining and periodic acid-Schiff （PAS） staining were used to examine morphological alterations in myocardial tissue. Masson’s trichrome staining was used to assess myocardial fiber alterations； Western blot analysis was used to measure myocardial tissue expression of transient receptor potential calcium channel 6 （TRPC6）， NOD-like receptor family pyrin domain-containing 3 inflammasome （NLRP3），absent in melanoma 2 inflammasome （AIM2）， Caspase-1， interleukin （IL）-6， and IL-1β in mouse myocardial tissue.Results　Compared with the WT control group， the WT+LPS group exhibited decreased cardiac EF （P<0.01）， FS （P<0.01）， and CO （P<0.05）， along with significantly increased myocardial tissue damage， glycoprotein deposition， and fibrosis （P<0.01）. Further analysis revealed that compared with the WT control group， the WT+LPS group exhibited markedly increased myocardial tissue expression of TRPC6， NLRP3， AIM2， Caspase-1， IL-6， and IL-1β （P<0.01）. Compared with the WT+LPS group， mice in the Trpc6-/- +LPS group exhibited elevated EF （P<0.01） and FS （P<0.05）， along with reduced myocardial tissue injury， glycoprotein deposition， and fibrosis （P<0.05）.Conclusion　Chronic LPS treatment can activate NLRP3/AIM2 inflammasomes through the up-regulation of TRPC6 expression， and then lead to chronic myocardial inflammatory injury and fibrosis， while Trpc6 knockdown can reduce myocardial inflammatory injury and fibrosis， and the mechanism is related to inhibiting the activation of NLRP3/AIM2 inflammasomes.

Abstract:
Objective　To investigate the role of interleukin （IL）-17A in acute inhalational pneumonia induced by the highly drug-resistant and hypervirulent Staphylococcus aureus strain USA300-R in mice.Methods　An acute inhalational pneumonia model was established in mice using an aerosolized pulmonary delivery technique. RNA sequencing （RNA-seq） and enzyme-linked immunosorbent assay （ELISA） were employed to examine the expression dynamics of Il17a mRNA and IL-17A protein， respectively， in the lungs of infected mice. Il17a knockout （Il17a-/-） mice were generated using CRISPR/Cas9 gene editing technology. The survival rate， body weight， bacterial load in lung tissue， and histopathological changes were compared between Il17a-/- and wild-type （WT） mice following inhalational infection with USA300-R.Results　12 hours after USA300-R infection， compared to pre-infection， the expression level of Il17a mRNA in lung tissue and the level of IL-17A protein in bronchoalveolar lavage fluid （BALF） increased by approximately 50-fold （P<0.01） and 6-fold （P<0.001）， respectively. Compared to WT mice， Il17a-/- mice exhibited approximately 10-fold higher bacterial loads in lung tissue at both 12 and 24 hours post-infection （P<0.001， P<0.05）. However， they showed significantly attenuated lung histopathological injury， reduced alveolar wall thickening， markedly decreased neutrophil infiltration， and an approximately 50% improvement in survival rate （P<0.05）.Conclusion　In acute Staphylococcus aureus USA300-R inhalational pneumonia， IL-17A contributes to bacterial clearance by recruiting neutrophils； however， excessive neutrophil infiltration exacerbates pulmonary inflammation and injury， reduces survival rates， and represents a potential therapeutic target.

Abstract:
Objective　To investigate the role of runt-related transcription factor 3 （RUNX3） in transforming growth factor-β1 （TGF-β1）-induced activation of mouse primary pulmonary fibroblasts （PFs）， and its effects on fibroblast activation protein （FAP） expression， cell proliferation， and collagen synthesis.Methods　PFs were isolated from C57BL/6 mice and cultured. A RUNX3 knockdown model was established using small interfering RNA （siRNA）. Cells were assigned to the control group （Control）， TGF-β1-treated group （TGF-β1）， negative control group （TGF-β1+siRNA-NC）， and RUNX3-silenced group （TGF-β1+si-RUNX3）. In addition， a RUNX3 overexpression rescue experiment was performed based on TGF-β1 stimulation. Protein and mRNA levels of RUNX3， FAP， and typeⅠcollagen （COL1A1） were measured by Western blot and reverse transcription quantitative real-time PCR （RT-qPCR）. Cell proliferation was assessed using CCK-8 and EdU assays. Co-expression of COL1A1 and FAP was examined by double immunofluorescence staining.Results　Compared with the Control group，RUNX3， FAP， and COL1A1 expression levels were upregulated in PFs in the TGF-β1 group （P<0. 01）. The CCK-8 assay showed that the absorbance value was reduced in the RUNX3 knockdown group compared with the negative control group（ P<0. 01）. Consistently， the EdU assay demonstrated a lower proportion of EdU-positive cells in the RUNX3 knockdown group than in the negative control group （P<0. 01）. Immunofluorescence double staining re‑vealed decreased fluorescence intensities of COL1A1 and FAP in the RUNX3 knockdown group relative to the nega‑tive control. Under RUNX3 overexpression conditions， these fluorescence signals exhibited a partial rebound（ P<0. 01）. Conclusion　 RUNX3 in TGF- β1-induced PFs may promote cell proliferation and collagen synthesis by positively regulating FAP expression. Targeting the RUNX3/FAP axis may represent a potential therapeutic strat‑egy for pulmonary fibrosis.

Abstract:
Objective　To investigate the effect of isoacteoside on the replication process of respiratory syncytial virus （RSV） and its underlying mechanism.Methods　RSV-infected HEp-2 cells （control group） were treated with isoacteoside （isoacteoside-treated group）. Real-time quantitative PCR （qPCR） was used to analyze the mRNA expression levels of the RSV fusion protein （F） gene and matrix protein （M） gene in both groups. Viral titers were determined by plaque assay to assess the inhibitory effect of isoacteoside on viral replication. The number of RSV inclusion bodies formed was determined using cell transfection and laser confocal imaging to evaluate the role of isoacteoside in the formation of inclusion bodies required for RSV replication. 18 seven-week-old female BALB/c mice were randomly divided into three groups： uninfected control group （n=6）， RSV-infected group （n=6）， and RSV-infected group treated with isoacteoside （n=6）. 5 days after infection， the mice were euthanized and lung tissues were collected for pathological analysis to assess the therapeutic effects of isoacteoside on RSV-induced lung tissue injury.Results　Compared to the control group， the isoacteoside-treated group exhibited the reduced mRNA levels of the RSV F gene （t=17.13， P<0.001） and M gene （t=18.22， P<0.001）， as well as a decrease in viral titer （t=15.32， P<0.001）. The number of inclusion bodies was also significantly reduced （t=16.12， P<0.001）. In the RSV-infected mouse experiment， compared to the RSV-infected group， the isoacteoside-treated group showed a decrease in the levels of inflammatory cytokines IL-4 （t=14.76， P<0.01） and IL-6 （t=21.13， P<0.001） in lung tissues， as well as decreased mRNA levels of the viral load-associated genes F （t=19.52， P<0.001） and M （t=18.76， P<0.001） in lung tissues. Pathological damage to lung tissue caused by RSV infection was alleviated.Conclusion　Isoacteoside inhibits RSV replication by interfering with the formation of RSV inclusion bodies and alleviates lung tissue injury induced by RSV infection in mice.

Abstract:
Objective　To explore the potential mechanisms by which Myristica fragrans prevents and treats atherosclerosis （AS）.Methods　The major active components of Myristica fragrans and their shared targets with AS were obtained from databases. The shared targets were subjected to pathway enrichment analysis and PPI network construction using the ClusterProfile package and the STRING database. Molecular docking between key targets and major active components was performed using AutoDock. Gene expression data from early and late， as well as stable and unstable AS plaques， were used to validate changes of key targets and major pathways during AS progression. Western blot， flow cytometry， YO-PRO-1/PI staining， and TUNEL staining were applied to verify the main mechanisms.Results　Nine active components of Myristica fragrans interacted with 293 AS-related targets， among which eight components acted on an average of 57.0% of the shared targets. Gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analyses indicated that the anti-AS effects mainly involved oxidative stress， inflammation， lipid metabolism， fluid shear stress， and apoptosis pathways. PPI network revealed JUN， CASP3， MAPK3， and AKT1 as key targets mainly involved in regulating apoptosis. Molecular docking showed stable binding conformations and high affinities between major components and these targets. Integrated analysis of gene expression in early and late， as well as stable and unstable AS plaques， showed significant enrichment of leukocyte apoptosis pathways in late and unstable plaques. Cell experiments further confirmed that Myristica fragrans significantly reduced Cleaved-CASP3（P=0.04）and p-MAPK3（P=0.000 3）levels， increased p-AKT1（P=0.004）levels， and inhibited macrophage apoptosis.Conclusion　Myristica fragrans potentially interferes with AS development by modulating pathways related to oxidative stress， inflammation， lipid metabolism， fluid shear stress， and apoptosis， with CASP3， MAPK3， and AKT1 serving as key targets mediating its anti-apoptotic and anti-AS effects.

Abstract:
Objective　To explore the inhibitory effect and related molecular mechanisms of Saponin of Schizocapsa plantaginea HanceⅠ （SSPHⅠ） on human nasopharyngeal carcinoma HONE-1 cells.Methods　The effect of SSPHⅠ on HONE-1 cell viability was detected using the CCK-8 assay. Its inhibitory effect on cell proliferation was evaluated through a colony formation assay. Changes in cell invasion ability were analyzed using the Transwell assay. Intracellular reactive oxygen species （ROS） levels were measured using the DHE fluorescent probe. The extent of intracellular content release was reflected by the LDH release assay. The rate of cell pyroptosis was detected using the Annexin-V/PI double staining method. Changes in the expression of proteins related to the classical pyroptosis pathway were examined by Western Blot.Results　CCK-8 assay showed that treatment with SSPHⅠ for 24 hours reduced HONE-1 cell viability in a concentration-dependent manner， with an IC50 value of 3.383 μmol/L. In the colony formation assay， the number of HONE-1 cell colonies gradually decreased with increasing concentrations of SSPHⅠ （P<0.01）. The Transwell assay revealed that the number of cells migrating through the chamber was reduced following SSPHⅠ treatment （P<0.01）. DHE fluorescence probe detection indicated that intracellular ROS fluorescence intensity increased after SSPHⅠ treatment （P<0.001）. The LDH release assay showed that LDH activity in the cell supernatant increased with higher concentrations of SSPHⅠ （P<0.001）. Annexin-V/PI double staining demonstrated that the proportion of Annexin-V/PI-positive cells increased after SSPHⅠ treatment （P<0.001）. Western blot analysis showed that， compared with the control group， the protein expression levels of cleaved-Caspase-1 and GSDMD-N-terminal were upregulated in SSPHⅠ-treated cells （P<0.05）， and NLRP3 protein expression levels also increased （P<0.05）. ELISA results showed that the levels of IL-1β and IL-18 in the cells increased with higher concentrations of SSPHⅠ （P<0.05）.Conclusion　SSPHⅠ can induce pyroptosis in nasopharyngeal carcinoma HONE-1 cells by regulating the ROS/NLRP3/Caspase-1 signaling axis， thereby exerting an anti-nasopharyngeal carcinoma effect. This suggests that SSPHⅠ may serve as a potential therapeutic agent for nasopharyngeal carcinoma.

Abstract:
Objective　To investigate the protective effect of genistein （Gen） on etoposide-induced chondrocyte senescence and its underlying mechanism.Methods　The C28/I2 cell line was treated with different concentrations of Gen and etoposide， and the cell viability was detected by the CCK-8 assay. The senescence model of C28/I2 chondrocytes was induced by etoposide， with Gen intervention. Senescence-associated β-galactosidase （SA-β-gal） staining was performed to detect the senescence-positive rate and staining characteristics of chondrocytes. The expressions of peroxiredoxin 6 （Prdx6）， cyclin-dependent kinaseto clarify the functional necessity of Prdx6.Results　Compared with the etoposide group， the C28/I2 chondrocyte viability significantly increased （P<0.01）， the expression ofsenescence-associated proteins p21 and p16 decreased （P<0.01， P<0.05）， the expression of senescence-associated genes p21 and p16 reduced （both P<0.01）， the fluorescence intensity of senescence-associated proteins p21 and p16 was diminished （P<0.05， P<0.01）， and the proportion of SA-β-gal-positive cells decreased （P<0.01） in the Gen+etoposide group. Compared with the Control group， the expression of Prdx6 was downregulated in the etoposide group （P<0.05）. Compared with the etoposide group， the expression of Prdx6 was upregulated in the Gen+etoposide group （P<0.01）. Compared with the Control group， the GPx activity significantly decreased in the si-Prdx6 group （P<0.01）. Furthermore， compared with the si-Prdx6 group， the GPx activity increased in the si-Prdx6+Gen group （P<0.05）. Molecular docking results revealed that Gen formed hydrogen bond interactions with the active site of Prdx6. After Prdx6 knockdown， the expression of senescence-associated genes p21 and p16 and the fluorescence intensity of senescence-associated proteins p21 and p16 both increased in the Gen+etoposide+si-Prdx6 group （both P<0.01）.Conclusion　Gen can inhibit etoposide-induced senescence of C28/I2 chondrocytes by upregulating the expression of Prdx6. This study provides potential drug targets and experimental basis for the prevention and treatment of chondrocyte senescence-related diseases.

Abstract:
Objective To explore the effects of transmembrane protein 16B （TMEM16B） on brain edema and blood-brain barrier after cerebral ischemia-reperfusion injury in rats. Methods TMEM16B overexpression and knockdown was performed by adeno-associated virus （AAV）， and then adult male Sprague-Dawley rats were sub⁃jected to middle cerebral artery occlusion （MCAO）. Rats were randomly divided into Sham group， MCAO group， AAV no-load group， TMEM16B overexpression group and TMEM16B knockdown group. Modified neurological se⁃verity scores， adhesive removal test and cylinder test were used to evaluate neurologic function. The ultrastructure of ischemic brain tissue was observed by transmission electron microscope. Brain water content was reflected by dry wet weight ratio of brain tissue. The expressions of TMEM16B， aquaporin4（AQP4）， Claudin5 and zonula oc⁃ cludens-1（ZO-1） were investigated by immunofluorescence and Western blot. Results Compared with the AAV no-load group， the sensory and motor functions of rats in TMEM16B overexpression group were significantly im⁃ paired. Mitochondria were swollen； mitochondrial cristae and tight junctions disappeared. The brain water content was higher in overexpression group. The expression of TMEM16B and AQP4 increased while the expression of Claudin5 and ZO-1 decreased （all P<0. 05）. Compared with the AAV no-load group， the rats in TMEM16B knock⁃down group showed some recovery in motor function. The mitochondrial cristae and structure were clear， and the basement membrane was partially blurred. The brain water content was lower in knockdown group. The protein lev⁃els of TMEM16B and AQP4 were lower while the levels of Claudin5 and ZO-1 were higher in TMEM16B knock⁃ down group than in AAV no-load group （all P<0. 05）. Conclusion An increase in TMEM16B expression aggra⁃vates brain edema and blood-brain barrier damage in rats after cerebral ischemia-reperfusion injury， while a de⁃ crease in TMEM16B expression alleviates brain edema and protects the blood-brain barrier.

Abstract:
Objective　To investigate the effects of salidroside （SAL） on the impaired bioactivity of endothelial progenitor cells （EPCs） in atherosclerotic （As） mice and the potential mechanisms regarding AMP-activated protein kinase （AMPK）.Methods　Atherosclerosis was induced in 8-week-old male ApoE-/- mice with high-fat diet. Intragastric administration of SAL was given to one mice group to investigate the effects of SAL on aortic plaque burden， plasma NO level， the migration and angiogenic capabilities of bone marrow-derived EPCs （BM-EPCs）. The proliferation， migration and vasculogenic properties of EPCs isolated from As mice were investigated in vitro. AMPK-sh-RNA or the AMPK inhibitor Compound C was used to investigate the role of AMPK/Akt/eNOS pathway in the regulatory effects of SAL.Results　Compared with As group， NO level was significantly elevated in SAL group. The sizes of atherosclerotic plaques at the aortic root were reduced with smaller lipid cores in SAL group compared with As group. Moreover， the migration and angiogenesis capacity of EPCs markedly decreased in As mice， while SAL treatment reversed these impairments. Incubation with SAL at concentrations of 20， 40， and 80 μmol/L for 48 hours significantly promoted the proliferation， migration， and angiogenesis of EPCs. AMPK-sh-RNA transfection abrogated the 20 μmol/L SAL improvement in EPC biological activities. Western blot analysis further demonstrated that treatment with Compound C blocked the activation of AMPK/Akt/eNOS signaling pathway induced by SAL.Conclusion　SAL upregulates the biological functions of EPCs through activating the AMPK/Akt/eNOS signaling pathway， thereby ameliorating EPC dysfunction during the pathological progression of atherosclerosis.

Abstract:
Objective　To investigate the functional role and underlying molecular mechanisms of fumarylacetoacetate hydrolase （FAH） in the progression of glioblastoma （GBM）.Methods　Differential expression analysis was performed on the TCGA-GBM， GSE4290， and GSE116520 datasets. Weighted gene co-expression network analysis （WGCNA） was used to identify key modules， and Cox regression and risk modeling were used to screen prognostic genes. Immune infiltration analysis of prognostic genes was carried out by using single-cell RNA sequencing panels. The clinical expression signature of FAH in GBM was analyzed in the TCGA and HPA databases. The functional role of FAH was validated by in vitro and in vivo experiments， and pathway analysis was performed to explore the underlying mechanisms.Results　A total of 152 overlapping genes were identified across the three GBM datasets （P<0.05）. WGCNA revealed that the turquoise module was most strongly associated with tumor purity， stromal score， immune score， and ESTIMATE score （P<0.001）. Compared with normal tissues， three prognostic genes （CTSD， FAH， and THBD） were upregulated in GBM and correlated with immune infiltration （P<0.05）. FAH mRNA and protein levels were elevated in GBM tissues relative to normal tissues， and its expression was significantly associated with age stratification and TP53 mutation （P<0.05）. CCK-8 assay results showed that， compared with the shNC group， the proliferative activity of GBM cells in the shFAH group was reduced （P<0.001）. Transwell migration and invasion assays demonstrated that， relative to the shNC group， the numbers of migrated and invaded cells in the shFAH group decreased （P<0.05）. Western blot analysis revealed that the protein expression levels of PI3K， p-AKT， and p-mTOR in the shFAH group decreased compared with those in the shNC group （P<0.05）. In vivo subcutaneous xenograft experiments further confirmed that tumor volume and weight significantly decreased in the shFAH group compared with the shNC group （P<0.001）.Conclusion　FAH promotes GBM progression by activating the PI3K/AKT/mTOR signaling pathway and may serve as a potential therapeutic target for GBM.

Abstract:
Objective　To investigate whether fullerene C60 administered by intratracheal spraying promotes N-bis （2-hydroxypropyl） nitrosamine （DHPN） induced-lung tumor development and the underlying mechanisms.Methods　A rat lung tumor model was induced by the DHPN drinking water method. Eight-week-old male Fisher 344 rats were randomly divided into 5 groups （n = 15）： DHPN+Veh group （0.2% DHPN + vehicle solution）， DHPN+C60-L group （0.2% DHPN + 250 μg/mL C60）， DHPN+C60-H group （0.2% DHPN + 500 μg/mL C60）， DHPN group （0.2% DHPN）， and C60-H group （normal drinking water + 500 μg/mL C60）. 0.2% DHPN in drinking water was given to initiate carcinogenesis， and 2 weeks later， different concentrations of fullerene C60 were administered by intratracheal spraying every two weeks for a total of 20 times. Two weeks after the last spraying， the incidence， number and size of lung tumors were statistically analyzed. In another mechanism animal experiment， 8-week-old male Fisher rats were divided into two groups with 5 rats in each group： C60 group and the control vehicle group. 500 μg/mL C60 or the control vehicle intratracheally sprayed every two days for a total of 5 times. Six hours after the last spraying， the lungs were removed and used for observation of C60 distribution and for detection of SOD， 8-OHdG and cytokines in lung tissues. Results　No lung tumorigenesis was observed in the C60-H group； the incidence， number and size of lung tumors in the DHPN+C60-H group were significantly higher than those in the DHPN+Veh group （P<0. 05， P<0. 001， P<0. 05） and the DHPN group （P<0. 01， P<0. 001， P<0. 01）， re‑spectively. In additional animal experiments designed for mechanistic investigation， the levels of SOD and 8- OHdG in the experimental group were markedly elevated compared with the control group （both P<0. 001）， with statistically significant differences. Conclusion　Intratracheal nebulization of high-concentration fullerene C60 pro‑ moted DHPN-induced lung tumorigenesis； C60 led to an increase in reactive oxygen species （ROS） production， thereby elevating the potential for gene mutation. This may represent one of the mechanisms underlying C60-in‑ duced promotion of lung tumorigenesis.

Abstract:
Objective　To explore the feasibility of constructing a programmed death receptor-1（PD-1） molecular probe for non-invasive micro-positron emission tomography/computed tomography （Micro-PET/CT） imaging of PD-1 protein in mouse S180 sarcoma.Methods　A transgenic PD-1 C57 S180 sarcoma mouse model was established using the S180 sarcoma cell injection. Furthermore， 124I-anti-PD-1 monoclonal antibody probe was synthesized. 18.5 MBq of the 124I-anti-PD-1 probe was injected into the tail vein of transgenic PD-1 C57 mice. Subsequently， S180 sarcoma was imaged using Micro-PET/CT.Results　Study successfully established a transgenic PD-1 C57 S180 sarcoma mouse model. Immunohistochemical （IHC） results showed PD-1 protein expression in S180 sarcoma. Micro-PET/CT imaging successfully visualized the PD-1 protein receptor in S180 sarcoma at different time points （20， 48， 72， and 120 h） after probe injection.Conclusion　The 124I-anti-PD-1 monoclonal antibody molecular probe successfully targets the PD-1 receptor in S180 sarcoma of transgenic PD-1 C57 mice， and presents clear Micro-PET/CT immunoassay results， thus it potentially enables the non-invasive screening of patients with PD-1 positive malignant tumors.

Abstract:
Objective　To explore the effect and mechanism of tumor necrosis factor alpha （TNF-α）-regulated circular RNA Williams Beuren syndrome chromosomal region 22 （circWBSCR22） on the malignant behaviors of cervical cancer cells.Methods　Human cervical cancer HeLa cells were treated with TNF-α. Reverse transcription quantitative polymerase chain reaction （RT-qPCR） was used to detect the expression level of circWBSCR22. Nuclear-cytoplasmic RNA separation experiments were conducted to determine the localization of circWBSCR22 in cells. Cell migration， invasion， viability， and proliferation were assessed through Transwell assay， tetrazolium salt （MTT） assay， and colony formation assay， respectively. Dual-luciferase reporter gene assays were performed to verify the targeting relationship between microRNA-512-5p （miR-512-5p） and circWBSCR22 or family with sequence similarity 60A （FAM60A）. RT-qPCR was used to detect the RNA levels of miR-512-5p and FAM60A. Western blot was used to detect the protein level of FAM60A.Results　Compared with the Control group， the expression level of circWBSCR22 in HeLa cells treated with TNF-α increased （P<0.05）. About 66% of circWBSCR22 was found in the cytoplasm. Compared with the control group， the migration （P<0.01） and invasion （P<0.01） of HeLa cells transfected with circWBSCR22 were enhanced， but the cell activity and cell proliferation were not significantly affected （P>0.05）. The double luciferase reporter gene assay showed that the relative luciferase activity of the co transfected circWBSCR22-WT and miR-512-5p groups was lower than that of the co transfected circWBSCR22-WT and miR-NC groups （P<0.01）. Compared with the control group， the expression level of miR-512-5p in HeLa cells transfected with circWBSCR22 decreased （P<0.01）. Compared with the control group， the migration （P<0.01） and invasion （P<0.05） ability of HeLa cells transfected with miR-512-5p decreased. Double luciferase reporter gene assay showed that compared with the co transfected FAM60A 3´-UTR-WT and miR-NC groups， the fluorescence activity of the co transfected FAM60A 3´-UTR-WT and miR-512-5p groups decreased （P<0.01）.Compared with the control group， the expression levels of FAM60A mRNA （P<0.05） and protein （P<0.01） in HeLa cells transfected with miR-512-5p decreased. Compared with the control group， the migration （P<0.01） and invasion （P<0.01） of cervical cancer HeLa cells transfected with FAM60A were enhanced. Compared with the control group， the expression levels of FAM60A mRNA （P<0.01） and protein （P<0.05） in the cells transfected with circWBSCR22 increased. The results of rescue experiment showed that compared with the pcDNA+miR-NC group， the expression levels of FAM60A RNA （P<0.01） and protein （P<0.001） in the pcDNA+miR-512-5p group decreased， and the ability of migration （P<0.01） and invasion （P<0.05） decreased； Compared with pcDNA+miR-512-5p group， the expression levels of FAM60A RNA （P<0.01） and protein （P<0.001） in circWBSCR22+miR-512-5p group increased， and the ability of migration （P<0.01） and invasion （P<0.05） was enhanced.Conclusion　TNF-α promotes the migration and invasion of cervical cancer cells through the circWBSCR22/miR-512-5p/FAM60A regulatory axis.

Abstract:
Objective　To examine the impact of Toll-like receptor 4 （TLR4）-mediated neutrophil extracellular traps （NETs） formation on isoproterenol （ISO）-induced heart failure （HF） in aged mice.Methods　The mice were randomly divided into Control group， Model group， TAK-242 （TLR4 inhibitor） group， LPS （TLR4 agonist） group and LPS+DNaseⅠ （NETs inhibitor） group， with 8 mice in each group. The HF model was established by continuous infusion of ISO through an osmotic pump， and TAK-242， LPS， and LPS+DNaseⅠ interventions were administered during the last week of infusion for 7 consecutive days. ELISA was used to detect the N-terminal pro-B-type natriuretic peptide （NT-pro BNP）， cardiac troponin I （cTnI）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， and serum and myocardial tissue myeloperoxidase-DNA （MPO-DNA） and neutrophil elastase-DNA （NE-DNA） levels. HE staining was used to observe the pathological changes of myocardial tissue. Masson staining was used to observe the deposition of collagen fibers in myocardial tissue. Immunofluorescence was used to observe the colocalization of Ly6G and MPO in myocardial tissue. The protein expressions of TLR4， MPO， NE and cit-H3 in myocardial tissue were detected by Western blot.Results　Compared with the Control group， the end-diastolic interventricular septal thickness （IVSD）， left ventricular end-diastolic diameter （LVEDD） and left ventricular end-systolic diameter （LVESD） in the Mode group significantly increased （P<0.01）， fractional shortening （FS） and ejection fraction （EF） significantly decreased （P<0.01）， and the myocardial structure was significantly damaged. The myocardial collagen fiber precipitation， serum NT-pro BNP， cTnI， IL-1β， IL-6 and TNF-α contents， MPO-DNA and NE-DNA contents in serum and myocardial tissue significantly increased （P<0.01）. The co-localization of Ly6G and MPO and the expression of TLR4， MPO， NE and cit-H3 proteins in myocardial tissue significantly increased （P<0.01）. After TAK-242 intervention， the above indicators were significantly reversed， while the injury was further aggravated in LPS group. DNaseⅠ treatment could partially alleviate the deterioration of myocardial structure and function induced by LPS.Conclusion　TLR4 and NETs are abnormally elevated in the myocardial tissue of aged HF mice. Treatment with the TLR4 inhibitor TAK-242 effectively suppresses NET formation， attenuates inflammatory responses， and enhances cardiac function in aged HF mice.

Abstract:
Objective　To evaluate the predictive value of epicardial adipose tissue （EAT） parameters for microvascular obstruction （MVO） formation in patients with ST segment elevation myocardial infarction （STEMI） using cardiac magnetic resonance quantification.Methods　A total of 139 STEMI patients were included in this study， and various parameters such as EAT thickness， volume， and mass index were measured utilizing cardiac magnetic resonance. All included patients were divided into MVO group and non-MVO group according to whether MVO occurred. Differences in EAT related parameters between two groups were compared and correlation analysis was applied to evaluate the correlation between quantitative indicators of EAT and indicators such as infarct size and ejection fraction. Logistic regression analysis was used to identify the relevant risk factors for MVO formation. Receiver Operating Characteristic （ROC） curve analysis was performed to assess the predictive value of the epicardial adipose tissue （EAT） quality index and other indicators for the occurrence of MVO.Results　Compared with non MVO group， patients in MVO group presented with higher peak troponin T levels， increase of neutrophil lymphocyte ratio （NLR） and C-reactive protein （CRP）， larger infarct size and compromised left ventricular ejection fraction （LVEF） （P<0.05）. Total EAT volume， EAT mass index， left atrioventricular EAT volume， left atrioventricular EAT mass index and thickness of EAT in the left atrioventricular groove were significantly higher in patients with MVO. Multivariate Logistic regression analysis demonstrated that NLR， peak troponin T levels and left atrioventricular EAT mass index were independent predictors of MVO. The ROC curve suggested that the left atrioventricular EAT mass index had the highest predictive power for MVO formation in STEMI patients.Conclusion　The parameters of EAT quantified by cardiac magnetic resonance serve as imaging biomarkers for predicting MVO formation in STEMI patients. These metrics enable risk stratification post-myocardial infarction and facilitate early identification of high-risk individuals， thereby supporting personalized therapeutic decision-making.

Abstract:
Objective　To explore the value of a multimodal MRI-based radiomics nomogram for differentiating human epidermal growth factor receptor-2 （HER-2） negative breast cancer molecular subtypes.Methods　A retrospective analysis was conducted on 190 patients with HER-2 negative breast cancer who underwent multimodal MRI examination， and the patients were divided into two molecular subtype groups： a HER-2 low expression group （n=108） and a HER-2 zero expression group （n=82）. The cases were randomly stratified and sampled at a ratio of 7∶3 and divided into a training set of 133 cases and a testing set of 57 cases. The clinical and radiological features of the patients were collected， the radiomics features based on T2-weighted imaging （T2WI）， diffusion-weighted imaging （DWI）， and dynamic contrast-enhanced （DCE）-MRI were extracted， and the clinical-radiological model， unimodal radiomics model， multimodal radiomics model， and combined model were constructed respectively. Then the nomogram combined multimodal radiomics signature （radsocre） with clinical-radiological features was used to construct a visualized predictive model， and the area under the curve （AUC） was used to compare the effectiveness of different models in distinguishing HER-2 low expression and zero expression subtypes.Results　A significant difference in radscore was demonstrated between the HER-2 low and HER-2 zero expression groups in both the training （P<0.000 1） and testing sets （P<0.01）. The AUC of the multimodal radiomics model in the training set and the testing set were 0.914 and 0.836， respectively， which was superior to any unimodal radiomics model. The nomogram demonstrated great diagnostic efficacy （AUC=0.930 in training set； AUC=0.865 in testing set）.Conclusion　A multimodal MRI-based nomogram incorporating radsocre and clinical-radiological features can accurately distinguish the subtypes of HER-2 negative breast cancer.

Abstract:
Objective　To investigate the fluctuations in serum uteroglobin-related protein 1（UGRP1） levels in Hashimoto’s thyroiditis （HT） patients before and after pregnancy， and to analyze the influencing factors.Methods　Ten healthy individuals and ten HT patients were enrolled. Thyroid fine needle aspiration cytology combined with immunohistochemistry was used to detect the expression of UGRP1 protein in thyroid cells between the two groups. A total of 30 healthy women were enrolled as the control group， and 149 HT patients were recruited， including 36 non-pregnant HT patients， 77 pregnant HT patients and 36 post-partum HT patients. According to levothyroxine sodium （L-T4） administration status， the pregnant HT group was further subdivided into the non-L-T4 subgroup （n=36） and the L-T4 subgroup （n=41）. Kruskal-Wallis H test was used to compare the general clinical data， thyroid-related indicators and serum UGRP1 levels among groups. Spearman correlation analysis and univariate linear regression analysis were performed to explore the influencing factors of serum UGRP1 levels in HT patients.Results　UGRP1 expression was negative in thyroid cells of all healthy individuals， while 80% of HT patients exhibited positive expression. The serum UGRP1 levels in the control group， non-pregnant HT group， pregnant HT group， and post-partum HT group were 359.52 （297.84， 440.60）， 695.77 （518.55， 865.04）， 207.96 （173.82， 264.91）， and 582.08 （280.83， 735.87） pg/mL， respectively， with statistically significant differences among groups （P<0.001）. Correlation analysis showed no significant correlation between serum UGRP1 levels and thyroid-related indicators or gestational age in pregnant HT patients. Univariate linear regression analysis revealed that pregnancy status was negatively correlated with serum UGRP1 levels in HT patients （β= -424.457， P<0.001）， while L-T4 administration had no statistically significant effect on serum UGRP1 levels in pregnant HT patients （P=0.890）.Conclusion　Serum UGRP1 levels are significantly higher in HT patients than in healthy individuals. Pregnancy is an important factor affecting serum UGRP1 levels in HT patients， which can lead to a decrease in UGRP1 levels， while L-T4 administration has no significant effect on serum UGRP1 levels.

Abstract:
Objective　To explore the clinical application value of CIE 1976 L*a*b*（Lab） color space in the differential diagnosis of early esophageal cancer and non-cancerous lesions.Methods　We selected the endoscopic images of patients with esophageal lesions confirmed by pathology who underwent white light imaging endoscopy （WLI） and narrow band imaging endoscopy （NBI）. Five regions of interest （ROI） were selected respectively from the mucosa of the lesion area and the mucosa of the surrounding normal area for labeling. The Lab color space parameters were extracted and counted， and the color difference values（ΔE*）were calculated.Results　A total of 213 eligible patients were included for analysis in the study. In WLI and NBI modes， there were differences in mucosal color between the early esophageal cancer group and the non-cancer group （P<0.05）. Compared with WLI mode， NBI mode could significantly increase the color difference between early esophageal cancer and non-cancerous lesions （P<0.05）. The lightness component value （L* value） of the early esophageal cancer lesion area was lower than that of the non-cancerous lesion area， and this color difference was more significant in the NBI mode （P<0.05）. In WLI mode， there was no significant difference in yellow-blue component value （b* value） between the mucosa of early esophageal cancer and non-cancerous lesions. However， in the NBI mode， the b* value of the mucosa in the non-cancerous lesion area was higher than that in the early esophageal cancer lesion area （P<0.05）. On the red-green axis， the mucosa of the early esophageal cancer and non-cancerous lesions was red in WLI mode and green in NBI mode. There was no significant difference in red-green component value （a* value） between the two groups.Conclusion　There are color differences between early esophageal cancer and non-cancerous lesions under WLI and NBI. The color of early esophageal cancer is darker under WLI， and the color of non-cancerous lesions is yellower under non-magnified NBI mode. Lab color space is helpful to identify early esophageal cancer and non-cancerous lesions.

Abstract:
Objective　To evaluate a novel nomogram based on contrast-enhanced MRI radiomics combined with clinical variables for the preoperative prediction of perineural invasion （PNI） in intrahepatic cholangiocarcinoma （ICC）.Methods　The clinical data of 59 ICC patients were retrospectively collected. According to postoperative pathology reports， the patients were divided into the non-PNI group （n = 33） and the PNI group （n = 26）. Regions of interest （ROI） were delineated from five MRI sequences. Radiomics features were then extracted and filtered to select those with the strongest discriminative power for PNI identification. These selected features were used to construct a radiomics model， which subsequently generated a quantitative radiomics score （radiomics score， Radscore）. Univariate analysis was applied to identify clinical variables associated with PNI， and the glm function was subsequently used to construct clinical and combined models. Finally， the models were evaluated using receiver operating characteristic （ROC） curves， calibration curves， and decision curve analysis （DCA）. The combined model was then visualized as a nomogram.Results　The clinical model included age， carbohydrate antigen 19-9 （CA19-9）， red blood cell distribution width， and albumin， whereas the Radscore included five radiomic features. The areas under the ROC curves （AUCs） for the clinical and radiomics models were 0.717 （95%CI： 0.586-0.848） and 0.896 （95%CI： 0.820-0.973）， respectively， whereas the combined model further improved its AUC to 0.917 （95% CI：0.848-0.987）. The calibration curves and DCA showed that the nomogram was well calibrated and provided the greatest net clinical benefit.Conclusion　The novel nomogram may serve as a basis for preoperative prediction of PNI status， thereby assisting clinical decision-making and guiding personalized treatment.

Abstract:
Objective　To analyze the etiological characteristics and risk factors of autosomal dominant polycystic kidney disease （ADPKD） complicated with urinary tract infection （UTI）.Methods　A retrospective analysis was performed on 244 patients with ADPKD admitted to the Department of Nephrology. The patients were stratified into a study group （n=138） and a control group （n=106） according to the presence or absence of concurrent UTI. All patients in the study group underwent etiological testing and drug susceptibility testing. Clinical data of both groups were collected to identify the risk factors for ADPKD complicated with UTI.Results　A total of 105 pathogens were detected in 138 cases in the study group， including 79 Gram-negative bacteria， 20 Gram-positive bacteria and 6 fungi. Escherichia coli showed a relatively high resistance rate to ampicillin， piperacillin and ciprofloxacin. Enterococcus faecalis had a relatively high resistance rate to tetracycline. There were statistically significant differ‑ences between the study group and the control group in terms of gender， history of urinary tract infection， C-reactive protein（ CRP）， increased urine red blood cells， increased urine white blood cells， increased percentage of neutrophils， blood sodium， blood calcium， total cholesterol， serum iron and transferrin saturation （P<0. 05）. Multivariate binary Logistic regression analysis revealed that female gender（ OR=2. 236， P=0. 004）， history of uri‑nary tract infection（ OR=2. 383， P=0. 002）， increased urine white blood cells（ OR=2. 726， P=0. 001）， increased percentage of neutrophils （OR=1. 040， P=0. 010）， elevated C-reactive protein （OR=1. 011， P=0. 007） and el‑evated blood calcium （OR=4. 389， P=0. 024） were risk factors of urinary tract infection in ADPKD patients （P<0. 05）. Conclusion　The main pathogens of UTI in patients with ADPKD are Gram-negative bacteria. Female， previous history of urinary tract infection， urinary leukocytosis， increased percentage of neutrophils， increased CRP and increased serum calcium are risk factors of UTI.

Abstract:
Objective　To analyze the risk factors for long-term cardiovascular events in patients undergoing long-term peritoneal dialysis （PD）， and to construct and validate a visual nomogram prediction model based on multiple parameters.Methods　A prospective cohort study was conducted， consecutively enrolling 248 maintenance PD patients （dialysis duration ≥ 3 months）. Demographic characteristics， clinical indicators， laboratory parameters， and echocardiographic indices （including left ventricular ejection fraction ［LVEF］， ratio of early diastolic mitral inflow velocity to early diastolic mitral annular velocity （E/e’）， etc.） were collected. The composite endpoint was defined as the occurrence of cardiovascular events or cardiovascular death， with non-cardiovascular death as the competing risk and loss to follow-up or the end of follow-up as censoring events. Fine-Gray competing risks model was used to screen independent predictors， based on which a nomogram model was constructed. Internal validation was performed using the Bootstrap method （1 000 resamplings）， and the concordance index （C-index） and time-dependent receiver operating characteristic （time-dependent ROC） curve were calculated to evaluate the model performance.Results　With a median follow-up of 29 months （interquartile range： 24–35 months）， 88 patients （35.48%） reached the composite endpoint， including 80 cases of cardiovascular events and 8 cases of cardiovascular death， and 4 patients died of non-cardiovascular causes. Multivariate Fine-Gray analysis revealed that age， diabetes mellitus， hemoglobin （HGB） level and E/e' ratio were independent influencing factors of the composite endpoint. Specifically， each 1-year increase in age was associated with a 3.0% increase in the risk of the composite endpoint （HR=1.030， P=0.006）； patients with diabetes mellitus had a 167.9% higher risk compared with non-diabetic patients （HR=2.679， P=0.007）； each 1g/L increase in HGB level contributed to a 1.5% reduction in the risk （HR=0.985， P=0.003）； and each 0.1 increase in E/e' ratio led to a 7.2% increase in the risk （HR=1.072， P=0.045）. The nomogram model had a C-index of 0.76 （95% CI： 0.698–0.820）， and the AUC of the time-dependent ROC curve reached 0.849 at 23 months of follow-up.Conclusion　Increased age， complicated with diabetes mellitus， decreased HGB， and elevated E/e' ratio are independent risk factors of long-term occurrence of cardiovascular events and cardiovascular death in patients undergoing long-term PD. The nomogram model constructed based on the above variables has good predictive value and clinical applicability， which can provide a reference for cardiovascular risk stratification and individualized intervention in long-term PD patients.

Abstract:
Objective　To evaluate the feasibility of various strength levels of deep learning image reconstruction （DLIR） algorithms for improving non-contrast abdominal CT image quality at ultra-low radiation doses， by comparing ultra-low-dose DLIR images with low-dose filtered back projection （FBP） images.Methods　A prospective collection of 85 patients undergoing non-contrast abdominal CT scans was performed， and a self-controlled study method was employed to conduct low-dose （LD） group and ultra-low-dose （ULD） group scans. The LD group used a noise index of 10 and employed FBP for image reconstruction （LD-FBP group）. The ULD group used a noise index of 30 and employed DLIR at different levels （low， medium， high）， resulting in three subgroups of reconstructed images： ULD-DLIR-L， ULD-DLIR-M， and ULD-DLIR-H. For each group， CT values， standard devia-tion（ SD）， signal-to-noise ratio（ SNR）， and contrast-to-noise ratio（ CNR） were measured and calculated for the liver， spleen， kidneys， aorta， psoas major， and subcutaneous fat. Effective dose（ ED） was also recorded. Two ra‑diologists independently performed subjective evaluations of image quality using a 5-point scale. Results　Com‑pared with the LD-FBP group， the ULD-DLIR-L group showed significantly lower SNR and CNR values in the liver， spleen， kidneys， aorta， and psoas major（ P<0. 001）， while the ULD-DLIR-H group exhibited significantly higher values（ P<0. 001）. The difference of SNR and CNR values for the ULD-DLIR-M group showed no statisti‑cally significant difference. For subjective evaluation， the scores of the ULD-DLIR-L and ULD-DLIR-M groups were lower than those of the LD-FBP group， while there was no statistically significant difference in scores between the ULD-DLIR-H group and the LD-FBP group. The ED value of the ULD group was approximately 88% lower than that of the LD group. Conclusion　Compared with the LD-FBP group， the ULD-DLIR-H group significantly reduces SD values while increasing SNR and CNR values， effectively improving the image quality of non-contrast abdominal CT scans.

Abstract:
Objective　To investigate the metabolic alterations of serum short chain fatty acids （SCFAs） in pediatric patients with atopic dermatitis （AD） and their correlation with different clinical phenotypes using targeted metabolomics.Methods　This study enrolled 87 AD patients and 67 healthy controls （HC）. Serum levels of eight SCFAs were quantified by ultra-high-performance liquid chromatography-mass spectrometry. The associations between SCFAs and AD were assessed using various statistical methods.Results　Compared with the HC group， levels of acetic acid （AA）， propionic acid （PA）， and caproic acid （CA） （P=0.002，P=0.002，P=0.043） decreased in the AD group. Logistic regression analysis identified AA （OR=0.449， 95% CI： 0.289–0.698） and PA （OR = 0.487， 95% CI： 0.324–0.732） as protective factors against AD. The combination of AA and PA yielded an area under the curve （AUC） greater than 0.7， indicating good diagnostic efficacy. Age-stratified analysis revealed that AA reduction was predominant in childhood， whereas PA reduction was predominant in adolescence. Pathway enrichment analysis showed significant enrichment of fatty acid biosynthesis （FDR=0.341， P=0.003） and vitamin K metabolism （FDR=1， P=0.039） pathways. Furthermore， subgroup analyses based on disease severity， personal/family history of atopy， and sex revealed no significant differences in SCFAs levels among the groups.Conclusion　Differential serum SCFAs and their enriched metabolic pathways may be implicated in the pathogenesis of AD.

Abstract:
Atherosclerosis （AS） is a chronic and inflammatory vascular disease. Macrophages are common immune cells and play an important role in the development of AS. In recent years， research has found that the formation of AS plaques is closely related to pathological and physiological processes such as macrophage polarization， energy metabolism， and lipid phagocytosis. This review aims to summarize the mechanism of macrophages in the development of AS， and to explore potential therapeutic methods for delaying AS by regulating macrophages， providing new ideas for the treatment and research of AS.

Abstract:
Neurodegenerative diseases such as Alzheimer's disease （AD）， Parkinson's disease （PD）， and Huntington's disease （HD） are increasingly becoming major challenges to global public health. The pathogenesis of these diseases is highly complex， with oxidative damage and ferroptosis emerging as potential core factors. Oxidative damage primarily results from the accumulation of reactive oxygen species （ROS） within cells， a process that impairs cellular function and viability. Ferroptosis， an iron-dependent form of programmed cell death， plays a crucial role in neuronal injury and apoptosis. Multiple studies indicate a significant interaction between oxidative damage and ferroptosis. This interplay not only accelerates the progression of neurodegenerative diseases but also offers new directions for potential therapeutic targets. This review systematically analyzes the mechanisms of oxidative damage and ferroptosis in various neurodegenerative diseases， evaluates current research frontiers， and explores feasible therapeutic strategies， aiming to provide new insights and perspectives for in-depth research in this field.

Abstract:
SMYD5 is a ribosomal methyltransferase with SET and MYND structural domains，which is a member of the SMYD family and is expressed in a variety of tissues，including ovary and testis. This enzyme participates in biological processes such as gene expression regulation，cell development and differentiation，and maintenance of genomic stability through ribosomal protein methylation modification. In recent years，research on SMYD5 has in⁃ creased in cancers including hepatocellular carcinoma，gastric adenocarcinoma，and lung cancer. Studies have re⁃ vealed that SMYD5 exhibits high expression levels in various diseases including hepatocellular carcinoma，gastric adenocarcinoma，lung cancer，and inflammatory bowel disease，influencing the progression of these conditions. This review summarizes the role of SMYD5 in hepatocellular carcinoma，inflammatory bowel disease，and other biological functions，aiming to provide a reference for related disease research.

Abstract:
Temporomandibular disorders （TMD） encompass a prevalent group of conditions affecting the masticatory muscles， temporomandibular joint， and associated structures. Currently， it is widely recognized by scholars both domestically and internationally that TMD has a complex etiology and pathogenesis. Most patients present clinically with pain， yet effective treatment options remain limited. In recent years， the role of ion channels in the pathological mechanisms and therapeutic approaches for TMD has garnered increasing attention. This review systematically summarizes the function of ion channels in the management of temporomandibular joint diseases. First， the multifaceted roles of ion channels in treating TMD are examined， including their regulatory effects on TMJ muscle dysfunction， joint structural abnormalities， and TMJ-related pain. Finally， the potential value of ion channel research in TMD therapy is discussed. With advancing research in this field， ion channel-targeted therapies for temporomandibular disorders are expected to achieve new breakthroughs， thereby providing an important theoretical foundation and practical guidance for developing novel clinical treatment strategies.