ObjectiveTo evaluate the anti-inflammatory activity of the novel magnolol ester derivative YW and to investigate its effects on chondrocyte senescence and preliminary mechanisms. MethodsMagnolol and p-methylbenzoic acid were used as raw materials to synthesize the magnolol ester derivative YW （Molecular Formula： C₂₆H₂₄O₃， Molecular Weight： 384.17， HPLC Purity >96%） via DCC/DMAP-catalyzed esterification. Cytotoxicity was assessed using the CCK-8 assay. A lipopolysaccharide （LPS）-induced RAW264.7 macrophage activation model and an interleukin-1β （IL-1β）-induced rat primary chondrocyte model were established. The release and mRNA expression of inflammatory factors including nitric oxide （NO）， IL-1β， tumor necrosis factor-alpha （TNF-α）， and IL-6 were detected by enzyme-linked immunosorbent assay （ELISA）， Griess reagent method， and quantitative real-time PCR （RT-qPCR）. The expression of senescence markers such as inducible nitric oxide synthase （iNOS）， pro-interleukin-1β （pro-IL-1β）， lysine acetyltransferase 7 （KAT7）， cyclin-dependent kinase inhibitor 1A （p21）， and cyclin-dependent kinase inhibitor 2A （p16）， as well as proteins related to chondrocyte extracellular matrix synthesis and catabolism， were analyzed by Western blot （WB）. Molecular docking was performed using Discovery Studio 2019 to validate target binding. ResultsYW exhibited no significant cytotoxicity at concentrations ≤20 μmol/L. YW concentration-dependently inhibited LPS-induced macrophage inflammatory cytokine release， significantly downregulated iNOS， Pro-IL-1β protein， and inflammatory cytokine mRNA expression （P<0.01）. YW stably bound to KAT7 protein （binding energy： -94.2 kcal/mol）； YW downregulated KAT7 and aging marker protein expression in naturally aged and IL-1β-induced chondrocyte models （P<0.01）； YW regulated chondrocyte matrix synthesis and catabolic protein expression in IL-1β-induced chondrocytes （P<0.01）. ConclusionYW inhibits macrophage activation and inflammatory cytokine release while downregulating KAT7 and senescence marker protein expression in chondrocytes， thereby blocking chondrocyte senescence.

Objective:To investigate the prognostic value of deep learning-based automated quantification of tumor-stroma ratio (TSR) in patients undergoing neoadjuvant therapy (NAT) for breast cancer.Methods:Specimens were collected from 209 breast cancer patients who received NAT at Renmin Hospital of Wuhan University from October 2019 to June 2023. TSR levels in pre-NAT biopsy specimens were automatically computed using a deep learning algorithm and categorized into low stroma (TSR≤30%), intermediate stroma (TSR 30% to ≤60%), and high stroma (TSR>60%) groups. Residual cancer burden (RCB) grading of post-NAT surgical specimens was determined to compare the relationship between TSR expression levels and RCB grades. The correlation of TSR with NAT efficacy was analyzed, and the association between TSR expression and patient prognosis was further investigated.Results:There were 85 cases with low stroma (TSR≤30%), 93 cases with intermediate stroma (TSR 30% to ≤60%), and 31 cases with high stroma (TSR>60%). Different TSR expression levels showed significant differences between various RCB grades ( P<0.05). Logistic univariate and multivariate analyses showed that TSR was a risk factor for obtaining a complete pathological remission from neoadjuvant therapy for breast cancer when it was used as a continuous variable ( P<0.05); COX regression and survival analyses showed that the lower the percentage of tumorigenic mesenchyme was, the better the prognosis of the patient was ( P<0.05). Conclusions:The deep learning-based model enables automatic and accurate quantification of TSR. A lower pre-treatment tumoral stroma is associated with a lower RCB score and a higher rate of pathologic complete response, indicating that TSR can predict the efficacy of neoadjuvant therapy in breast cancer and thus holds prognostic significance. Therefore, TSR may serve as a biomarker for predicting therapeutic outcomes in breast cancer neoadjuvant therapy.

Objective To explore the shared mechanisms of genes related to three common chronic diseases non-alcoholic fatty liver disease(NAFLD),type 2 diabetes mellitus(T2DM)and atherosclerosis(AS)with ageing as well as potential therapeutic agents by using bioinformatics analysis,machine learning algorithms and molecular docking methods and techniques.Methods Ageing-related genes were collected and organized from AgeingAtlas,CellAge,GenAge,and MSigDB databases.After taking the intersection of genes related to NAFLD,T2DM and AS obtained from databases such as CTD,DisGeNET,GeneCards,OMIM,PharmGKB,and TTD and the gene sets obtained based on the GEO differential gene analysis,we obtained the set of related disease genes for these three common chronic diseases.The KEGG pathway enrichment analysis was performed on the ageing gene set and the three disease-related gene sets using the clusterProfiler package,and the intersection was taken.The enriched genes in the KEGG pathway were merged and imported into the STRING database;the PPI network was constructed.We analyzed the core sub-modules in the PPI network using the MCODE tool and calculated the essential values of Nim and Cim for each node and module.Meanwhile,three machine learning models were used to screen the feature genes:the Lasso regression model,the Boruta algorithm,and the random forest model.The HIT2.0 database was utilized to find the targeted TCM small molecules related to the key feature genes.Small molecules were evaluated and analyzed by ADMET using SwissADME and ADMETlab 3.0 online system.The molecular docking method was utilized to dock the key action targets and screen small molecules.Results A total of 1 325,616,78,and 597 genes related to ageing,NAFLD,T2DM,and AS were obtained.The KEGG pathway enrichment analysis results for ageing and the three diseases were taken to intersect to get two shared intersecting pathways containing 243 genes.The PPI network was constructed,and Cluster 2 had the highest Cim value among the three core submodules.According to the results of signature gene screening,combined with PPI network module analysis results,four signature genes related to ageing were found:CDK6,CDKN1A,MYC,and PTEN.These four targets have 94 potential TCM small molecule candidates,among which resveratrol(RSV)is a TCM small molecule common to these four targets.The ADMET evaluation showed that it had good drug-forming properties.The PTEN target had a high Nim value,and molecular docking of RSV with PTEN showed good binding stability.Conclusion A potential herbal small molecule,RSV,was identified from the perspective of ageing,which may prevent and treat three common chronic diseases,namely,NAFLD,T2DM and AS,by regulating the key gene PTEN.

Objective:To investigate the prognostic value of deep learning-based automated quantification of tumor-stroma ratio (TSR) in patients undergoing neoadjuvant therapy (NAT) for breast cancer.Methods:Specimens were collected from 209 breast cancer patients who received NAT at Renmin Hospital of Wuhan University from October 2019 to June 2023. TSR levels in pre-NAT biopsy specimens were automatically computed using a deep learning algorithm and categorized into low stroma (TSR≤30%), intermediate stroma (TSR 30% to ≤60%), and high stroma (TSR>60%) groups. Residual cancer burden (RCB) grading of post-NAT surgical specimens was determined to compare the relationship between TSR expression levels and RCB grades. The correlation of TSR with NAT efficacy was analyzed, and the association between TSR expression and patient prognosis was further investigated.Results:There were 85 cases with low stroma (TSR≤30%), 93 cases with intermediate stroma (TSR 30% to ≤60%), and 31 cases with high stroma (TSR>60%). Different TSR expression levels showed significant differences between various RCB grades ( P<0.05). Logistic univariate and multivariate analyses showed that TSR was a risk factor for obtaining a complete pathological remission from neoadjuvant therapy for breast cancer when it was used as a continuous variable ( P<0.05); COX regression and survival analyses showed that the lower the percentage of tumorigenic mesenchyme was, the better the prognosis of the patient was ( P<0.05). Conclusions:The deep learning-based model enables automatic and accurate quantification of TSR. A lower pre-treatment tumoral stroma is associated with a lower RCB score and a higher rate of pathologic complete response, indicating that TSR can predict the efficacy of neoadjuvant therapy in breast cancer and thus holds prognostic significance. Therefore, TSR may serve as a biomarker for predicting therapeutic outcomes in breast cancer neoadjuvant therapy.

Objective To explore the shared mechanisms of genes related to three common chronic diseases non-alcoholic fatty liver disease(NAFLD),type 2 diabetes mellitus(T2DM)and atherosclerosis(AS)with ageing as well as potential therapeutic agents by using bioinformatics analysis,machine learning algorithms and molecular docking methods and techniques.Methods Ageing-related genes were collected and organized from AgeingAtlas,CellAge,GenAge,and MSigDB databases.After taking the intersection of genes related to NAFLD,T2DM and AS obtained from databases such as CTD,DisGeNET,GeneCards,OMIM,PharmGKB,and TTD and the gene sets obtained based on the GEO differential gene analysis,we obtained the set of related disease genes for these three common chronic diseases.The KEGG pathway enrichment analysis was performed on the ageing gene set and the three disease-related gene sets using the clusterProfiler package,and the intersection was taken.The enriched genes in the KEGG pathway were merged and imported into the STRING database;the PPI network was constructed.We analyzed the core sub-modules in the PPI network using the MCODE tool and calculated the essential values of Nim and Cim for each node and module.Meanwhile,three machine learning models were used to screen the feature genes:the Lasso regression model,the Boruta algorithm,and the random forest model.The HIT2.0 database was utilized to find the targeted TCM small molecules related to the key feature genes.Small molecules were evaluated and analyzed by ADMET using SwissADME and ADMETlab 3.0 online system.The molecular docking method was utilized to dock the key action targets and screen small molecules.Results A total of 1 325,616,78,and 597 genes related to ageing,NAFLD,T2DM,and AS were obtained.The KEGG pathway enrichment analysis results for ageing and the three diseases were taken to intersect to get two shared intersecting pathways containing 243 genes.The PPI network was constructed,and Cluster 2 had the highest Cim value among the three core submodules.According to the results of signature gene screening,combined with PPI network module analysis results,four signature genes related to ageing were found:CDK6,CDKN1A,MYC,and PTEN.These four targets have 94 potential TCM small molecule candidates,among which resveratrol(RSV)is a TCM small molecule common to these four targets.The ADMET evaluation showed that it had good drug-forming properties.The PTEN target had a high Nim value,and molecular docking of RSV with PTEN showed good binding stability.Conclusion A potential herbal small molecule,RSV,was identified from the perspective of ageing,which may prevent and treat three common chronic diseases,namely,NAFLD,T2DM and AS,by regulating the key gene PTEN.

Objective : To establish an interleukin-1β (Il-1β) induced inflammatory model of rat articular chondro- cytes (ACs) , and to investigate the relationship between the expression of lysine acetyltransferase 7 (KAT7) under inflammatory stimulation and the senescence of ACs. Methods: Primary ACs were obtained by digestion of rat knee cartilage with collagenase type Ⅱ and identified. The inflammatory model of ACs was induced by IL-1β . KAT7 was over-expressed or knocked down in ACs by adeno-associated virus infection or small interfering RNA transfection , respectively. A negative control group was set up. Transwell assay was used to detect cell migration ability. Senes- cent cells were stained with senescence-associated β-galactosidase (SA-β-Gal) . Western blot ( WB) was used to detect the protein expression levels of KAT7 , collagen type II (Col Ⅱ ) , matrix metalloproteinase 13 (MMP13) , tumor protein p53 (p53) and cyclin-dependent kinase inhibitor 1A (p21) . The cells of negative control group and KAT7 over-expression group were performed for RNA sequencing , and WB was used to verify the related signaling pathways obtained by Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. Results: Compared with the control group , the SA-β-Gal staining was enhanced , the protein expression of Col Ⅱ decreased , the pro- tein expression of MMP13 and p53 increased , the cell migration ability decreased , and the expression of KAT7 also increased in the ACs of rats after IL-1β stimulation. Compared with the negative control group , the SA-β-Gal stai- ning was enhanced , the protein expression of Col Ⅱ decreased , the protein expression of MMP13 , p53 and p21 in- creased , and the cell migration ability decreased in the KAT7 over-expression group. Compared with the negative control group , the SA-β-Gal staining was weakened , the protein expression of Col Ⅱ increased , the protein expres- sion of MMP13 , p53 and p21 decreased , and the cell migration ability was enhanced in the KAT7 knockdown inflammatory model of ACs. KEGG enrichment analysis showed that phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway was activated. Compared with the negative control group , the relative protein ex⁃pression levels of phosphorylated protein kinase B (p⁃AKT)/AKT and phosphorylated mammalian target of rapamy⁃cin (p⁃mTOR)/mTOR in KAT7 over⁃expression group increased. The relative protein expression levels of p ⁃AKT/AKT and p ⁃mTOR/mTOR in KAT7 knockdown cells decreased. Conclusion Rat ACs with high expression of KAT7 exhibit senescence and osteoarthritis phenotype , and the mechanism may be related to the activation of PI3K/AKT/mTOR signaling pathway by KAT7.

It is necessary to explore potent therapeutic agents via regulating gut microbiota and metabolism to combat Parkinson's disease(PD).Dioscin,a bioactive steroidal saponin,shows various activities.How-ever,its effects and mechanisms against PD are limited.In this study,dioscin dramatically alleviated neuroinflammation and oxidative stress,and restored the disorders of mice induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).16 S rDNA sequencing assay demonstrated that dioscin reversed MPTP-induced gut dysbiosis to decrease Firmicutes-to-Bacteroidetes ratio and the abundances of Enterococcus,Streptococcus,Bacteroides and Lactobacillus genera,which further inhibited bile salt hy-drolase(BSH)activity and blocked bile acid(BA)deconjugation.Fecal microbiome transplantation test showed that the anti-PD effect of dioscin was gut microbiota-dependent.In addition,non-targeted fecal metabolomics assays revealed many differential metabolites in adjusting steroid biosynthesis and pri-mary bile acid biosynthesis.Moreover,targeted bile acid metabolomics assay indicated that dioscin increased the levels of ursodeoxycholic acid,tauroursodeoxycholic acid,taurodeoxycholic acid and β-muricholic acid in feces and serum.In addition,ursodeoxycholic acid administration markedly improved the protective effects of dioscin against PD in mice.Mechanistic test indicated that dioscin significantly up-regulated the levels of takeda G protein-coupled receptor 5(TGR5),glucagon-like peptide-1 receptor(GLP-1R),GLP-1,superoxide dismutase(SOD),and down-regulated NADPH oxidases 2(NOX2)and nu-clear factor-kappaB(NF-κB)levels.Our data indicated that dioscin ameliorated PD phenotype by restoring gut dysbiosis and regulating bile acid-mediated oxidative stress and neuroinflammation via targeting GLP-1 signal in MPTP-induced PD mice,suggesting that the compound should be considered as a prebiotic agent to treat PD in the future.

Objective:To explore the risk factors of recurrence after resection of thigh soft tissue malignant tumors and the evaluation of the postoperative lower limb function.Methods:A total of 211 patients who underwent thigh soft tissue malignant tumor resection in the Department of Orthopaedics of the second affiliated Hospital of Medical College of Zhejiang University from May 2011 to May 2021 were retrospectively analyzed, including 117 males and 94 females, aged 53 (43, 65) years. There were 59 cases of atypical/well-differentiated liposarcoma, 30 cases of other types of liposarcoma, 33 cases of malignant fibrous histiocytoma, 19 cases of fibrosarcoma, 12 cases of rhabdomyosarcoma, 12 cases of leiomyosarcoma, 9 cases of synovial sarcoma, and 37 cases of others. The tumor involvement of muscle, bone, blood vessel, nerve tissue and intermuscular compartment, postoperative Musculoskeletal Tumor Society (MSTS) functional score and recurrence were recorded. The relationship between anatomical structure and postoperative recurrence was analyzed by Cox logistic regression to determine risk factors; the cumulative recurrence rate of different compartment groups (medial compartment, anterior compartment, and posterior compartment) were compared; The Box plot was used to compare the postoperative function of patients with resection of different anatomical structures (e.g., vastus lateralis, biceps femoris, femur, etc.).Results:A total of 34 out of 211 patients relapsed, with a recurrence rate of 16.1%. The recurrence time ranged from 2.6 months to 91.6 months after operation, with a median recurrence time of 37.0 (18.2, 52.8) months. Three independent risk factors were found to be associated with recurrence, namely: pathological grade [ HR=3.86, 95% CI(1.75, 8.51)], involvement of vastus intermedius [ HR=3.05, 95% CI(1.53, 6.06)], and involvement of vastus medialis [ HR=3.17, 95% CI(1.56, 6.41)]. The recurrence rate of patients with anterior chamber tumor resection was 35.3%, which was higher than that of patients without tumor resection (16.2%), and the difference was statistically significant ( P=0.020). There was no significant difference in recurrence rate between patients with medial chamber tumor resection and patients without tumor resection ( P>0.05). The recurrence rate of patients with posterior compartment tumor resection was 12.3%, which was lower than that of unresectable patients (37.6%), and the difference was statistically significant ( P=0.002).The postoperative MSTS score of 167 patients averaged 26±3.2 points (9-30 points). After intraoperative resection of part of the femur, vastus intermedius, vastus medialis, and rectus femoris, the patient's function was relatively poor [corresponding median MSTS score: 25 (23, 28), 26 (24 28), 26 (24,28), 26(24, 27)]. Conclusion:The risk factors for local recurrence after resection of thigh soft tissue malignant tumors include: pathological grade of the tumor, and whether the vastus intermedius or vastus medialis is involved. Anterior compartment tumors have a higher recurrence rate after surgery. If the tumor involves the above-mentioned anatomical structures, more attention should be paid to the risk of local recurrence after surgery.

Objective:To explore the clinical characteristics of dislocation after resection and reconstruction of tumors involving pelvic area II, and to try to propose intraoperative and postoperative techniques to prevent its occurrence.Methods:From March 2011 to March 2021, 122 patients with resection and reconstruction involving pelvic area II were retrospectively analyzed. Among them, 17 had postoperative dislocation, 32 had pelvic area II, and 31 had pelvic area I+II. There were 40 cases in the pelvic area II+III, and 19 cases in the pelvic area I+II+III. There were 49 female patients and 73 male patients; the mean age was 47 years (9-73 years). The Musculoskeletal Tumor Society (MSTS) score of lower extremity patients after reduction was evaluated, and the clinical characteristics of dislocation, such as dislocation direction, dislocation time and reduction method, were counted and analyzed, and feasible prevention measures were analyzed based on the imaging characteristics after resection and reconstruction. dislocation method.Results:Among the 122 patients, there were 17 cases of dislocation, and the dislocation rate was 13.7%. Among them, 12 cases were anterior dislocation, accounting for 70.6% of anterior dislocation; 5 cases were posterior dislocation, and the proportion of posterior dislocation was 29.4%, difference (χ 2=4.52, P=0.033). There were 12 cases of dislocation within 3 months after operation, accounting for 70.6%; 1 case of dislocation occurred in 5 months after operation, and the other 4 cases of dislocation occurred for more than one year. The MSTS score of the dislocation patients after reduction was 56.1±15.6% (20%-80%). Combined with postoperative imaging examinations, the feasible methods for preventing dislocation include: The anteversion angle of the acetabular cup can be appropriately reduced (0°-10°); The acetabular cup can be appropriately shifted to the rear of the rotation center; Appropriately reducing the anteversion angle of the femoral neck prosthesis, the combination of the three can reduce the occurrence of femoral-acetabular prosthesis impingement; at the same time, the affected limb is controlled in a neutral position with strict nail shoes after surgery to avoid the occurrence of external rotation. Conclusion:The dislocation after tumor resection and reconstruction in pelvic area II is mostly anterior dislocation. Postoperative CT scan of the pelvis shows that the space between the neck of the femoral stem prosthesis and the lower edge of the acetabulum is small, which is prone to impingement. If the anteversion angle of the acetabular cup is appropriately reduced, the acetabular prosthesis is appropriately displaced posteriorly, and the anteversion angle of the femoral neck is reduced by 5°, the possibility of femoral-acetabular impingement can be reduced, thereby reducing the risk of postoperative dislocation.

OBJECTIVETo explore the short-term effects of ambient PM10 pollution on mortality due to circulatory diseases, and to study the modifying effect of season on the association between ambient PM10 concentration and mortality in Nanjing.

METHODSDaily mortality, air pollution, and meteorological data from 1 January 2009 to 31 December 2013 in Nanjing were collected; Time-stratified case-crossover design was used to analyze the associations between daily average concentration of inhalable particle (PM10) and mortality due to circulatory diseases; Odds ratios (OR) and 95% confidence intervals (CI) were calculated; And stratified analysis was conducted to compare the mortality risks of circulatory diseases exposed to outdoor PM10 in warm season (May-October) with that in cool season (November-April).

RESULTSThe mean daily average concentrations of PM10, NO2, and SO2 from 2009 to 2013 in Nanjing were 109.1, 51.5, and 35.4 µg/m(3), respectively. During our study period, a total of 78 299 circulatory disease deaths were recorded. On average, there were approximately 43 circulatory disease deaths per day; and deaths due to cardiac diseases, ischemic heart disease and cerebrovascular diseases were 19, 10 and 24 per day, respectively. After adjusting for the meteorological variables such as daily temperature, relative humidity and air pressure, a10 µg/m(3) increment of PM10 was associated with the mortality on values of (OR) 1.002 4 (95% CI: 1.000 1-1.004 8) for total circulatory diseases, 1.004 8 (95% CI: 1.001 3-1.008 3) for cardiac diseases, 1.007 3 (95% CI: 1.002 4-1.012 3) for ischemic heart disease and 1.000 5 (95% CI: 0.998 5-1.002 4) for cerebrovascular diseases, respectively. In summer season, the OR values of daily mortality for total circulatory diseases and cerebrovascular diseases were 1.008 0 (95% CI: 1.003 9-1.012 2) and 1.005 7 (95% CI: 1.002 0-1.009 4), respectively, the corresponding OR values in cool season were 1.001 4 (95% CI: 0.998 5-1.004 2) and 0.998 8 (95% CI: 0.996 4-1.001 2), respectively; The effect estimates in warm season were higher than that in cool season.

CONCLUSIONThe elevated levels of ambient PM10 were positively associated with the increase of mortality due to circulatory diseases, and season may modify the effects of outdoor PM10 pollution on mortality.

Air Pollutants ; adverse effects ; Air Pollution ; Cardiovascular Diseases ; mortality ; Cerebrovascular Disorders ; mortality ; China ; Cross-Over Studies ; Humans ; Particulate Matter ; adverse effects ; Seasons ; Temperature