Background Toluene and chlorobenzene have been designated as surrogate contaminants in the challenge test for evaluating the safety of recycling processes for food-contact recycled polyethylene terephthalate (rPET). Establishing a reliable analytical method is essential for ensuring the compliant use of rPET and safeguarding food safety. Objective To develop a rapid quantitative method for determining toluene and chlorobenzene in rPET using headspace gas chromatography-mass spectrometry (HS-GC-MS), as part of the challenge test for process safety evaluation. Methods The effects of different chromatographic columns and headspace conditions on detection of target analytes were investigated. Three columns HP-5 ms UI (30 m×0.25 mm×0.25 μm), DB-624 (30 m×0.32 mm×1.8 μm), and VF-WAXms (30 m×0.25 mm×0.25 μm) were compared for separation efficiency and peak shape. Headspace equilibration temperatures (50-100 ℃) and equilibration times (10-30 min) were evaluated to determine the optimal instrumental parameters. The effect of sample grinding on recovery was assessed to select the best pretreatment conditions. The established method was validated for selectivity, linearity, sensitivity, accuracy, and precision, and was subsequently applied to the analysis of 12 rPET samples. Results The target analytes achieved good separation and response within 15 min, under the optimized conditions using an HP-5 ms UI column, a headspace equilibration temperature of 60 ℃ and a 10 min equilibration time. Direct analysis without grinding yielded satisfactory recovery rates. Toluene and chlorobenzene showed excellent linearity (0.0030-5.0 mg·kg⁻¹, R²≥0.999). The limits of detection (LOD) and quantification (LOQ) were 0.00003 mg·kg⁻¹ and 0.00011 mg·kg⁻¹ for toluene respectively, and 0.00011 mg·kg⁻¹ and 0.00037 mg·kg⁻¹ for chlorobenzene respectively. The recoveries of the matrix spike at low and high spiking levels ranged from 88.6% to 110%, with relative standard deviations of 0.52%-4.9% (n=6). Among the 12 rPET samples from different recycling stages, three samples contained detectable levels of toluene (0.196-0.250 mg·kg⁻¹) and chlorobenzene (0.704-0.867 mg·kg⁻¹). Conclusion The proposed method is simple, rapid, highly sensitive, and accurate, making it suitable for determining toluene and chlorobenzene in food-contact rPET. It provides a robust technical approach for the safety evaluation of rPET recycling processes.

ObjectiveTo explore the biological mechanism of bone loss caused by perfluorooctanoic acid (PFOA) through transcriptomic analysis, and to provide new insights into regulating perfluoroalkyl substances (PFAS) applications and the prevention of hazards affecting bone health. MethodsMouse embryonic osteoblast precursor cells (MC3T3-E1) were exposed to 0.1, 1, 10, and 100 μmol·L^-¹ PFOA for 24 hours to assess the effects on cell viability and alkaline phosphatase (ALP) activity, and to determine the critical concentration of PFOA toxicity. The transcriptome sequencing (RNA-seq) was performed to identify differentially expressed genes (DEGs) induced by PFOA. Gene ontology (GO) analysis and gene set enrichment analysis (GSEA) were conducted to identify significantly affected gene pathways. Additionally, Seahorse XF metabolic phenotyping and reverse transcription polymerase chain reaction (RT-PCR) were used to validate the key pathways. ResultsExposure to 10 and 100 μmol·L^-¹ PFOA significantly reduced the cell viability and ALP activity of MC3T3-E1 cells. Therefore, the results of transcriptomic analysis for 10 μmol‧L^-1 PFOA exposure found that a total of 80 DEGs were identified, including 32 upregulated genes and 48 downregulated genes. According to GO analysis, PFOA mainly affected cellular components such as mitochondrion and nucleus, molecular functions involving GTPase activity and GTP binding, as well as biological process related to mRNA processing. GSEA identified the downregulation of the β-oxidation of fatty acid pathway in mitochondria. Metabolic phenotyping reserches showed that PFOA indeed reduced mitochondrial aerobic respiration capacity and adenosine triphosphate (ATP) production, and the ratio of ATP production from cellular aerobic respiration to glycolysis was significantly decreased as well. The mRNA expression of glucose metabolism-related genes (GK, G6PD, and CS), as well as fatty acid metabolism-related genes (CPT1A and CPT2), were significantly downregulated. ConclusionPFOA reduces bone formation by inhibiting energy metabolism and β-oxidation of fatty acid pathways in osteoblasts, whihc lays the foundation for revealing the mechanism of PFOA exposure induced bone loss.

OBJECTIVES: To investigate the effects of quercetin on cuproptosis and L-type calcium currents in the myocardium of diabetic rats. METHODS: Forty SD rats were randomized into control group and diabetic model groups. The rat models of diabetes mellitus (DM) induced by high-fat and high-sugar diet combined with streptozotocin (STZ) injection were further divided into DM model group, quercetin treatment group, and empagliflozin treatment group (n=10). Blood glucose and body weight were measured every other week, and cardiac function of the rats was evaluated using echocardiography. HE staining, Sirius red staining, and wheat germ agglutinin (WGA) analysis were used to observe the changes in myocardial histomorphology, and serum copper levels and myocardial FDX1 expression were detected. In cultured rat cardiomyocyte H9c2 cells with high-glucose exposure, the effects of quercetin and elesclomol, alone or in combination, on intracellular CK-MB and LDH levels and FDX1 expression were assessed, and the changes in L-type calcium currents were analyzed using patch-clamp technique. RESULTS: The diabetic rats exhibited elevated blood glucose, reduced body weight, impaired left ventricular function, increased serum copper levels and myocardial FDX1 expression, decreased L-type calcium currents, and prolonged action potential duration. Quercetin and empagliflozin treatment significantly lowered blood glucose, improved body weight, and restored cardiac function of the diabetic rats, and compared with empagliflozin, quercetin more effectively reduced serum copper levels, downregulated FDX1 expression, and enhanced myocardial L-type calcium currents in diabetic rats. In H9c2 cells, high glucose exposure significantly increased myocardial expressions of FDX1, CK-MB and LDH, which were effectively lowered by quercetin treatment; Elesclomol further elevated FDX1, CK-MB and LDH levels in the exposed cells, and these changes were not significantly affected by the application of quercetin. CONCLUSIONS Quercetin ameliorates myocardial injury in diabetic rats possibly by suppressing myocardial cuproptosis signaling and restoring L-type calcium channel activity.
Animals ; Quercetin/pharmacology* ; Calcium Channels, L-Type/metabolism* ; Diabetes Mellitus, Experimental/metabolism* ; Rats, Sprague-Dawley ; Rats ; Myocytes, Cardiac/drug effects* ; Myocardium/pathology* ; Male

Objective:To explore the short- and medium-term clinical outcomes of covered stent implantation in the treatment of transplant renal artery stenosis (TRAS).Methods:A retrospective analysis was conducted on the clinical data of 12 consecutive patients with TRAS (transplant renal artery stenosis) who underwent covered stent implantation in Henan Provincial People′s Hospital from January 2021 to December 2024. The changes in indicators such as serum creatinine (Cr), blood urea nitrogen (BUN), mean arterial pressure (MAP), peak systolic velocity (PSV), intrarenal resistance index (RI), and the diameter of the stenotic site were analyzed before the operation, one week after the operation, six months after the operation, and 12 months after the operation. Repeated measures analysis of variance was used to investigate the changes of each observation index over time.Results:The surgery was successfully performed on all 12 patients, with a technical success rate of 100%. Among them, 8 cases used self-expanding covered stents, and 4 cases used balloon-expandable covered stents. One week, six months and twelve months after the surgery, the levels of Cr, BUN, PSV and MAP were all lower than those before the surgery. The RI and the diameter of the stenotic site were significantly increased compared with those before the surgery, the difference was statistically significant ( P<0.05). During the perioperative period and the postoperative follow-up period, no surgery-related complications were found. Conclusion:The implantation of the covered stent can effectively relieve the stenosis of the transplant renal artery, significantly improve renal function, and reduce blood pressure levels in TRAS patients, while maintaining excellent short-to medium-term clinical outcomes.

Objective:To identify the main components of Huanglian Jiedu Decoction(HLJDD)using ultra-high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS),and to explore the potential mechanism of action of HLJDD in the treatment of gouty arthritis(GA)using network pharmacology and molecular docking methods.Methods:We identi-fied the chemical components of HLJDD by combining UPLC-Q-TOF-MS data acquired in both positive and negative ion modes with reference standards,relevant literature,and database searches.We analyzed the potential therapeutic mechanism of HLJDD for GA by using network pharmacology to determine the intersection targets between the active ingredients of HLJDD and GA for further enrich-ment analysis and visual network mapping.The binding affinity of the active ingredients with the intersection targets was validated through molecular docking.Results:A total of 47 components were identified by UPLC-Q-TOF-MS;54 key components of HLJDD for GA treatment and 37 intersection targets were determined by net-work pharmacology;and the top 10 key targets by Degree value were obtained by protein-protein interaction analysis.The Gene On-tology functional enrichment analysis revealed 20 biological pro-cesses,7 cellular components,and 8 molecular functions.The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis demonstrated 96 GA-related intervention pathways,in which inflammatory signaling pathways such as interleukin-17(IL-17)and tu-mor necrosis factor(TNF)were involved.Molecular docking verified that the key components of HLJDD had high binding affinity with the core targets.Conclusion:The identified key components in HLJDD,such as phellodendrine,coptisine,wogonin,and β-sitosterol,may alleviate GA by regulating multiple core targets in the IL-17 and TNF pathways,such as PTSG2,which provides a theoretical ba-sis for future investigation into the mechanism of action of HLJDD.

ObjectiveTo explore the role of flurochloridone (FLC) on osteogenic differentiation and the potential mechanism of inhibiting bone formation, and to provide new insights into bone health risks associated with FLC pesticide exposure. MethodsNeonatal rat skull differentiation primary osteoblast model was used to investigate the effects of 1, 10 and 100 μmol·L^-1 FLC exposure on cell viability, osteogenic differentiation alkaline phosphatase (ALP) activity, and bone mineralization nodule formation, respectively. The potential mechanism underlying the inhibition of FLC on osteoblast differentiation was analyzed using osteogenic differentiation gene chip technique, and the expression of key genes and proteins in the pathway was validated using reverse transcription polymerase chain reaction (RT-PCR) and protein immunoblotting (Western blot) methods. ResultsExposure to FLC at a concentration of 100 μmol·L⁻¹ reduced cell proliferation, ALP activity, and the formation of mineralized nodules in primary osteoblasts. Gene chip analyses revealed that exposure to 10 μmol·L⁻¹ FLC induced 15 differentially expressed genes (DEGs). Among these, MMP9 and Tnf were up-regulated, while Nkx3⁃2, Tuft1, Bmp2, Col12a1, Pparg, Enam, Igf1, Bmp5, Bmp3, Calcr, Egf, Igfbp3, and Col14a1 were down-regulated. Results of protein-protein interaction analyses and gene ontology enrichment analyses indicated that FLC inhibited the BMP/SMAD pathway involved in osteogenic differentiation. FLC suppressed the protein expression of BMP2 and Osterix, as well as the expression of key genes critical for osteogenic differentiation and ossification, such as BMP2, Runx2, SMAD1, and SMAD5 in the BMP/SMAD pathway. ConclusionFLC affects osteogenic differentiation and bone formation potential by regulating the BMP/SMAD axis and the expression of osteogenic genes, suggesting its potential risk in bone metabolism.

Objective:To investigate the predictive value of radiomics features derived from dynamic contrast-enhanced MRI (DCE-MRI) based on habitat imaging technology for pathological complete response after neoadjuvant therapy (NAT) for breast cancer.Methods:All patients were female, aged 25-67 years. Patients were stratified into training ( n=83) and validation ( n=36) sets via stratified random sampling (7∶3 ratio). Pathological complete remission (pCR) and non-pathological complete remission (non-pCR) were defined using the Miller-Payne grading system. All patients underwent DCE-MRI before NAT. ITK-Snap software was used to outline the region of interest (ROI), the imaging histological features of the entire tumor region were extracted and screened, a traditional imaging histological model for predicting post-NAT pCR (ROI overall model) was constructed; the tumor region was divided into three subregions using habitat imaging technology, and the imaging histological features within ROI subregion 1, ROI subregion 2, and ROI subregion 3 were extracted and screened, and the habitat imaging model for predicting post-NAT pCR were constructed (ROI subregion 1 model, ROI subregion 2 model, ROI subregion 3 model). Univariate logistic regression identified clinical predictors of pCR for clinical model construction. Combined models integrating clinical predictors and habitat imaging features were established. The efficacy of each model in predicting pCR after NAT in breast cancer was evaluated using receiver operating characteristic curves and area under the curve (AUC), and the efficacy of clinical application of the models was evaluated using decision curve analysis (DCA). Results:Of the 119 patients, 74 were pCR patients, with 52 in the training set and 22 in the validation set, and 45 were non-pCR patients, with 31 in the training set and 14 in the validation set. Logistic regression analysis showed that human epidermal growth factor receptor 2 status ( OR=0.254, 95% CI 0.093-0.697, P=0.008) was an independent predictor of pCR after NAT, and this was used to construct a clinical prediction model. The predictive efficacy of ROI subregion 1 model and ROI subregion 2 model in the habitat model was higher than that of the traditional imaging histology model (ROI overall model), with AUCs of 0.805, 0.748,0.728 for the training set and 0.776,0.718,0.708 for the validation set, respectively. The combined clinical prediction model for predicting pCR after NAT in breast cancer had AUCs of 0.877 and 0.818 for the training and validation sets, respectively. DCA showed a higher net benefit for the combined model than for the traditional imaging histology model and the habitat imaging histology model. Conclusion:Compared with the traditional method of extracting the entire tumor region, extracting radiomics features from DCE-MRI subregions based on habitat imaging technology can improve the predictive performance of NAT efficacy in breast cancer.

Objective To investigate the clinical value of baseline 18F-fluorodeoxyglucose(18F-FDG)PET/CT metabolic parame-ters and certain clinical indicators in predicting the prognosis of patients with diffuse large B-cell lymphoma(DLBCL).Methods A retrospec-tive analysis was conducted on the baseline 18 F-FDG PET/CT data of 73 DLBCL patients who received R-CHOP treatment.Patients were divided into progression group(24 cases)and non-progression group(49 cases)based on disease progression within 2 years post-treatment.The lesion maximum standardized uptake value(SUVmax),tumour-to-liver blood pool SUVmax ratio(LLR)and tumour-to-mediastinal blood pool SUVmax ratio(L-BPR)were analyzed using receiver operating characteristic(ROC)curves.Kaplan-Meier(K-M)survival curves analysis were performed based on the optimal thresholds of SUVmax,LLR and L-BPR.x2 tests were used to analyze and compare the relationship between each parameter and disease progression.Indicators that were significant in the x2 tests were included in the multivariate Cox regression analysis.Results The area under the curve(AUC)for LLR,L-BPR,and SUVmax were 0.920,0.914,and 0.848,respectively,with optimal thresholds of 7.41,10.67,and 14.70.Based on these thresholds,K-M survival curves analysis showed that the 2-year progression-free survival(PFS)rates for DLBCL patients were 79.2％vs 30.8％(P＜0.001),74.3％vs 22.0％(P=0.009),and 79.5％vs 51.6％(P=0.002),respectively.Significant differences were observed between the progression and non-progression groups in terms of LLR,L-BPR,SUVmax,extranodal involvement,international prognostic index(IPI)score,lactate dehydrogenase(LDH)level,Eastern Cooperative Oncology Group(ECOG)score,and β2-microglobulin(β2-MG)levels(P＜0.05).Multivariate Cox regression analysis revealed that the IPI score and LLR were independent predictors affecting the 2-year PFS of DLBCL patients(P＜0.05).Conclusion Baseline 18F-FDG PET/CT metabolic parameter LLR and IPI score are inde-pendent factors for predicting the prognosis of DLBCL patients.

Objective:To explore the short- and medium-term clinical outcomes of covered stent implantation in the treatment of transplant renal artery stenosis (TRAS).Methods:A retrospective analysis was conducted on the clinical data of 12 consecutive patients with TRAS (transplant renal artery stenosis) who underwent covered stent implantation in Henan Provincial People′s Hospital from January 2021 to December 2024. The changes in indicators such as serum creatinine (Cr), blood urea nitrogen (BUN), mean arterial pressure (MAP), peak systolic velocity (PSV), intrarenal resistance index (RI), and the diameter of the stenotic site were analyzed before the operation, one week after the operation, six months after the operation, and 12 months after the operation. Repeated measures analysis of variance was used to investigate the changes of each observation index over time.Results:The surgery was successfully performed on all 12 patients, with a technical success rate of 100%. Among them, 8 cases used self-expanding covered stents, and 4 cases used balloon-expandable covered stents. One week, six months and twelve months after the surgery, the levels of Cr, BUN, PSV and MAP were all lower than those before the surgery. The RI and the diameter of the stenotic site were significantly increased compared with those before the surgery, the difference was statistically significant ( P<0.05). During the perioperative period and the postoperative follow-up period, no surgery-related complications were found. Conclusion:The implantation of the covered stent can effectively relieve the stenosis of the transplant renal artery, significantly improve renal function, and reduce blood pressure levels in TRAS patients, while maintaining excellent short-to medium-term clinical outcomes.

Objective To investigate the clinical value of baseline 18F-fluorodeoxyglucose(18F-FDG)PET/CT metabolic parame-ters and certain clinical indicators in predicting the prognosis of patients with diffuse large B-cell lymphoma(DLBCL).Methods A retrospec-tive analysis was conducted on the baseline 18 F-FDG PET/CT data of 73 DLBCL patients who received R-CHOP treatment.Patients were divided into progression group(24 cases)and non-progression group(49 cases)based on disease progression within 2 years post-treatment.The lesion maximum standardized uptake value(SUVmax),tumour-to-liver blood pool SUVmax ratio(LLR)and tumour-to-mediastinal blood pool SUVmax ratio(L-BPR)were analyzed using receiver operating characteristic(ROC)curves.Kaplan-Meier(K-M)survival curves analysis were performed based on the optimal thresholds of SUVmax,LLR and L-BPR.x2 tests were used to analyze and compare the relationship between each parameter and disease progression.Indicators that were significant in the x2 tests were included in the multivariate Cox regression analysis.Results The area under the curve(AUC)for LLR,L-BPR,and SUVmax were 0.920,0.914,and 0.848,respectively,with optimal thresholds of 7.41,10.67,and 14.70.Based on these thresholds,K-M survival curves analysis showed that the 2-year progression-free survival(PFS)rates for DLBCL patients were 79.2％vs 30.8％(P＜0.001),74.3％vs 22.0％(P=0.009),and 79.5％vs 51.6％(P=0.002),respectively.Significant differences were observed between the progression and non-progression groups in terms of LLR,L-BPR,SUVmax,extranodal involvement,international prognostic index(IPI)score,lactate dehydrogenase(LDH)level,Eastern Cooperative Oncology Group(ECOG)score,and β2-microglobulin(β2-MG)levels(P＜0.05).Multivariate Cox regression analysis revealed that the IPI score and LLR were independent predictors affecting the 2-year PFS of DLBCL patients(P＜0.05).Conclusion Baseline 18F-FDG PET/CT metabolic parameter LLR and IPI score are inde-pendent factors for predicting the prognosis of DLBCL patients.