OBJECTIVE: To investigate effect of low-dose cadmium exposure in early life on liver function and lipid metabolism in a mouse model for Alzheimer's disease(AD mice). METHODS: AD and wild-type(WT) mice were exposed to cadmium via drinking water(0,1,and 10 mg/L) from one week before pregnancy until 6 months of age. Blood samples were collected from F1 generation mice at the ages of 1,3,and 6 months,and serum lipid and liver function indices were measured. Liver tissues were collected,and mRNA expression levels of metallothionein 1(MT1) and metallothionein 2(MT2) in the livers were determined by quantitative real-time polymerase chain reaction(RT-qPCR). RESULTS: Compared with 3-month-old toxicant-exposed WT mice, age-and dose-matched AD mice showed increased body weight, liver weight,liver coefficient,TC,TG,HDL,and ALT(P<0.05),along with decreased LDL,AI,and hepatic MT1 mRNA expression(P<0.05). In comparison with 6-month-old toxicant-exposed WT mice,age-and dose-matched AD mice exhibited elevated body weight,liver weight,TC,TG,and HDL(P<0.05),along with decreased AST and AST/ALT ratio(P<0.05). Furthermore,interactive effects of cadmium and APP/PS1 genotype were observed on body weight, liver weight, TC, LDL, and TG/HDL ratio(P<0.05). CONCLUSION: Low-dose cadmium exposure in early life can induce dyslipidemia and abnormal liver function in AD mice,therefore exposure to environmental cadmium levels(1 mg/L) can reduce expression level of MT1 and MT2.

OBJECTIVE: To establish a phosgene-induced acute lung injury（ALI） model in SD rats and to investigate mechanisms for improvement effect of captopril on ALI rats. METHODS: Forty male SD rats were randomly divided into control,phosgene poisoning（Phos）,Captopril（CAP） and combined groups with phosgene and captopril（Phos+CAP）, with ten rats in each group. Rats dynamically inhaled 41 mg/m³ phosgene for 30 minutes in Phos and Phos+CAP groups and received intraperitoneal injection with 5 mg/kg Captopril immediately after phosgene poisoning in CAP and Phos+CAP groups. Survival rates of the rats were determined within 24 hours after administration. Lung function of the rats was measured using a whole-body plethysmography（WBP）system after 24 hours. Rat lung tissues were collected to measure the lung coefficient. HE staining of lung tissues was performed to assess pathological damage. Total protein concentration was determined in bronchoalveolar lavage fluid（BALF） by BCA method. Inflammatory factors including IL-1β and TNF-α were detected by ELISA in serum, BALF and lung homogenate. Activities of superoxide dismutase（SOD） and glutathione peroxidase（GPx）,content of malondialdehyde（MDA） were measured by kits. Levels of Angiotensin-converting enzyme（ACE）and angiotensin Ⅱ（Ang Ⅱ） were determined by immunohistochemistry and ELISA. Apoptosis in lung tissues was observed by TUNEL staining. Expression of Bcl-2 in lung tissue was detected by Western blotting. RESULTS: Compared with the control group, the 24 h survival rate of rats in the phosgene exposure group was 80%.Pulmonary hemorrhage spots were observed,and HE staining showed typical pathological changes of acute lung injury. Levels of IL-1β and TNF-α in bronchoalveolar lavage fluid,lung homogenate,and serum were elevated（P<0.01）. Activities of SOD and GPx were decreased（P<0.05）,while MDA content as well as ACE and Ang Ⅱ levels were significantly increased（P<0.01）. Apoptotic cell density was markedly increased（P < 0.01） along with reduced expression of Bcl-2 protein（P<0.01）. Compared with Phos group,levels of IL-1β and TNF-α and the activity of angiotensin-converting enzyme（ACE） and content of angiotensin Ⅱ（Ang Ⅱ） were significantly decreased in the Phos+CAP group（P<0.05）. The number of apoptotic cells decreased（P<0.05）,and Bcl-2 protein expression declined（P<0.01）. CONCLUSION: Captopril significantly alleviated acute lung injury which was induced by phosgene. Alleviation mechanisms may be related to alveolar barrier repair, enhanced antioxidant defense and inflammation suppression. This study provides experimental evidence for a clinical approach of phosgene poisoning.

OBJECTIVE: To explore mechanisms by which acteoside(ACT) ameliorates sarcopenia,and to suggest a scientific basis for treatment of sarcopenia with natural products. METHODS: Fifty 6-week-old SPF male C57 BL/6 mice were randomly divided into five groups:a negative control group(normal saline),a model group(5 mg/kg dexamethasone via intraperitoneal injection), a positive control group(5 mg/kg dexamethasone plus 0.37 mg/kg perindopril erbumine),a dexamethasone plus 50 mg/kg ACT group,and a dexamethasone plus100 mg/kg ACT group. All mice were administered intragastrically once daily for 4 consecutive weeks. Before sacrifice,fasting blood glucose concentrations were measured using a blood glucose meter. Serum levels of tumor necrosis factor-α(TNF-α) were determined by ELISA. After sacrifice, gastrocnemius muscle tissues were harvested, and glycogen deposition in skeletal muscle was evaluated by Periodic Acid-Schiff(PAS) staining.Protein expression levels of matrix metalloproteinase-9(MMP-9) in skeletal muscle were detected by immunohistochemistry. RESULTS: Compared with the negative control mice, dexamethasone-induced sarcopenia in mice exhibited impaired glucose tolerance(P<0.01),and serum levels of the pro-inflammatory factor TNF-αwere significantly elevated(P<0.01). Compared with the 50 mg/kg ACT intervention group,glucose tolerance was partially restored in the 100 mg/kg ACT intervention group. Compared with the model group, TNF-αconcentrations were decreased in both ACT intervention groups(P<0.01). Compared with the negative control mice,model mice showed markedly reduced glycogen deposition between muscle fibers,with sparse distribution and weakened staining intensity of magenta granules. Additionally, MMP-9 protein expression was abnormally increased and widely distributed in the intermuscular fiber spaces. In contrast, the ACT intervention groups exhibited increased density of glycogen granule deposition and significantly reduced intensity of MMP-9 positive signals. Among them, the PAS staining intensity and MMP-9 distribution pattern in the 100 mg/kg ACT intervention group were restored to near-normal levels. CONCLUSION: ACT regulated blood glucose concentration in vivo in a dose-dependent manner. Mechanistically, it ameliorated pathological progression of sarcopenia by inhibiting TNF-α-and MMP-9-mediated inflammatory responses and excessive degradation of the extracellular matrix. These findings provide a pharmacological basis for development of natural product-based drugs in prevention and treatment of sarcopenia.

OBJECTIVE: To investigate changes in cellular stemness transition in radiation-induced skin injury in cell culture. METHODS: Human immortalized keratinocytes(Ha Ca T) were exposed to X-ray at 0,5,10,and 20 Gy. Cell migration ability at 24 h and 48 h post-irradiation was assessed using the scratch wound healing assay and transwell migration assay. Western blot was used to detect the expression of stemness-related proteins KLF4,c-myc and LGR5 in Ha Ca T cells at 24 h and 48 h after irradiation. In the animal experiments,the skin of the right hind limb of SD rats was irradiated with a single dose of 20 Gy X-ray. At 15 and 35 days post-irradiation, western blot was performed to detect the expression of stemness-related proteins SOX2,Nanog, KLF4, c-myc, and LGR5 in the irradiated skin cells. A non-irradiated control group(0 Gy) was included, with 4 rats per group. RESULTS: Compared with the control group(0 Gy), the in vitro scratch wound healing assay at 48 h post-irradiation showed that cell migration ability was significantly reduced in the5, 10, and 20 Gy groups(P<0.05). In contrast, the transwell assay demonstrated significantly enhanced migration ability in all irradiated groups(P<0.01), with the 10 Gy group still retaining a certain level of migration capacity. At 24 h post-irradiation,KLF4,c-myc and LGR5 protein expression levels in Ha Ca T cells were significantly decreased in all irradiated dose groups(P<0.01). At 48 h post-irradiation,KLF4,c-myc and LGR5 protein expression levels were significantly increased(P<0.01). In the animal experiments,compared with the control group(0 Gy), at 15 days after 20 Gy X-ray irradiation, expression levels of stemness-related proteins SOX2, c-Myc, and LGR5 in rat skin were significantly elevated(all P<0.01). At 35 days postirradiation, expression levels of SOX2, Nanog, and LGR5 were significantly decreased(P<0.05 or P<0.01).CONCLUSION: In radiation-induced skin injury, cellular stemness transition capacity expressed dynamic changes which might cause inhibition of repair,promotion of regeneration with gradual decline in the long term.

OBJECTIVE: To investigate changes in differentially expressed genes in peripheral blood after in vitro exposure to different doses of⁶⁰Co γ rays and to identify new biomarkers for radiation damage.METHODS: Peripheral blood from four healthy adult volunteers were irradiated with different doses of⁶⁰Co γrays（0, 2, 4, 6, and 8 Gy）. Total RNA samples were extracted from irradiated blood using RNA high-throughput sequencing（RNA seq）. The sequencing data was analyzed using bioinformatics methods. The R language software was used to screen differentially expressed genes between groups, and a common set of differentially expressed genes was identified. In addition, GO functional enrichment analysis and KEGG pathway enrichment analysis were performed on the differentially expressed genes using the Metascape platform.A protein interaction network was constructed using STRING database and key genes were screened using the Cytoscape software. RESULTS: A total of 281 common and differentially expressed genes were screened,including 89 lncRNAs and 192 mRNAs. The enrichment of these expressed genes was shown by GO to mainly included lymphocyte activation, positive regulation of immune effector process, positive regulation of immune response, inflammatory response,regulation of cellular component size. The KEGG pathway was enriched in cytokine cytokine receptor interactions, viral carcinogenesis, Th17 cell differentiation, ribosomes, T cell receptor signaling pathways,platelet activation,and intercellular adhesion junctions. Protein-protein interaction network screening identified 10 key genes: CD80, CD40, IL2 RA, CCR7, NFKB1, TP53, IL21R, STAT6,PDCD1,and SLAMF1. CONCLUSION: The 10 key genes might be useful biomarkers for demonstration of radiation damage.

OBJECTIVE: Radiomics and machine learning show great potential in oncology prediction and prognosis. This study aimed to develop a machine learning model integrating contrast-enhanced CT radiomics features and clinical characteristics to predict lymph node metastasis in cervical squamous cell carcinoma(CSCC) patients. METHODS: In this retrospective study, patients with pathologically confirmed CSCC who underwent radical hysterectomy at the Cancer Hospital of Shantou University Medical College between January 2016 and December 2021 were enrolled. Preoperative contrast-enhanced CT images obtained within three weeks before surgery were collected. A Radscore was calculated by delineating tumor volumes and extracting/selecting radiomic features. Clinical characteristics were also collected. Four machine learning modelsLogistic regression, LDA, SVM, and Naïve Bayes-were built using the clinical features and Radscore to identify the optimal predictive model. RESULTS: The Naive Bayes model demonstrated the best and most stable overall performance,achieving an AUC of 0.957. The AUCs for the other models were 0.953 for SVM,0.943 for Logistic regression, and 0.941 for LDA. On the test set, the Naive Bayes model also achieved excellent accuracy(0.819), sensitivity(0.915), specificity(0.727), and an F1-score(0.889). SHAP analysis identified CA-125 as the most important predictor in the model's decision-making. CONCLUSION: We successfully developed and validated a high-performance Naïve Bayes model for predicting lymph node metastasis risk in CSCC patients. SHAP interpretability analysis further confirmed CA-125,CEA,FIGO stage,and Radscore as key predictors,enhancing the model's transparency and clinical credibility.

OBJECTIVE: This study aimed to investigate liver toxicity of the organophosphate flame retardant triethyl phosphate(TEP) on zebrafish(Danio rerio), and to provide a scientific basis for the ecological risk assessment of TEP. METHODS: Four-month-old AB strain wild-type zebrafish were exposed to environmentally relevant concentrations of TEP(0, 1, 10, 100, and 1 000 μg/L) for 28 days. After exposure,body weight and body length were measured. Histopathological damage of the liver was observed via hematoxylin-eosin(HE) staining. Liver function indicators(activities of ALT and AST), oxidative stress and inflammation-related indicators(gene expression levels and enzyme activities),metabolic enzyme(CYP450),and expression levels of key molecules in the Keap1-Nrf2-ARE signaling pathway were detected using RT-qPCR,ELISA,and other methods. RESULTS: TEP exposure had no significant effect on the body weight or body length of zebrafish. HE pathological sections of exposed groups showed sinusoidal dilation and edema in the liver;nuclear lysis was observed in the 1 and 10 μg/L TEP groups,while nuclear lysis,nuclear pyknosis,and anuclear areas were simultaneously observed in the 100 and 1 000 μg/L groups. Activities of ALT and AST were significantly elevated. The mRNA expression of oxidative stress-related genes(Sod1,Sod2,Gstp1.1,Gstp1.2, Cat, Cyp1a, Cyp3a, and Gpx1a) were downregulated, while the mRNA expression of proinflammatory factors(IL-1β,TNF-α and IL-6) was upregulated. The activities of SOD,CAT,GSH-Px,and CYP450 were significantly decreased,whereas MDA content was significantly increased. For key molecules in the Keap1-Nrf2-ARE signaling pathway:the mRNA expression of Keap1 was upregulated,while Nrf2,Ho1,and NQO1 expressions were downregulated. Meanwhile, the protein expressions of Keap1, Nrf2 and NQO1 were significantly downregulated. CONCLUSION: Environmentally relevant concentrations of TEP induced oxidative stress injury accompanied by inflammatory response in zebrafish liver by interfering with the Keap1-Nrf2-ARE signaling pathway. The results provide scientific reference for ecological prevention and control of TEP pollution in water bodies.

OBJECTIVE: To investigate expressions of cytokeratin 7（CK7） and cytokeratin 19（CK19） in primary liver cancer and to analyze their clinical significance. METHODS: Immunohistochemical Envision method was used to detect expression of cytokeratins in cancer tissues from 56 liver cancer patients. Spearman correlation was used to calculate the correlation coefficient between cytokeratins. The chi-square test or Fisher's exact probability test was used to analyze relationships between expression of CK7 and CK19 proteins and clinicopathological characteristics of patients. ROC curve was used to analyze accuracy of CK7 and CK19 protein expression levels in predicting the survival status of patients. Kaplan-Meier method was used to calculate survival rates of patients,and the Log-rank test was performed. RESULTS: The positive expression rates of CK7, CK8, CK18, CK19, and CK20 proteins in liver cancer tissues were 30.36%, 89.29%,69.64%,21.43%,and 5.36%,respectively. The expression levels of CK7 and CK19 proteins were positively correlated（r=0.602,P=0.00）. In primary liver cancer, expression of CK7 protein was related to the degree of differentiation and pathological type of liver cancer tissue（χ²=8.155,12.118,respectively,P<0.05）. Expression of CK19 protein was related to tumor diameter,tissue differentiation,tumor stage,and pathological type（χ²=7.263,15.955,7.292,10.672,respectively,all P<0.05）. Prognostic analysis showed that the area under the receiver operating characteristic（ROC） curve for CK7 protein in predicting the survival status of patients was0.750,with a standard error of 0.210 and a 95% confidence interval of（0.338,1.000）. The area under the ROC curve for CK19 was 0.875,with a standard error of 0.155 and a 95% confidence interval of（0.571,1.000）. CK8, CK18, and CK20 showed low predictive accuracy for patient survival status. Survival analysis results showed that CK7 and CK19 had no significant effect on the survival time of patients（P>0.05）. CONCLUSION: Expressions of CK7 and CK19 were highly and positively correlated. Both CK7 and CK19 affected tissue differentiation and pathological type of liver cancer patients. Prognostic analysis showed that expression of CK7 and CK19 had good prediction on postoperative survival status of patients. However,survival analysis showed that they have no significant effect on postoperative survival time of patients.

OBJECTIVE: To investigate correlations between protein expression of immunoglobulin heavy constant gamma 1(IGHG1) and thymosin β10(TMSB10) with pathological complete response(p CR) and Miller-Payne(MP) grading following neoadjuvant therapy(NAT) in patients with human epidermal growth factor receptor 2(HER2)-positive breast cancer, and to evaluate their predictive value. METHODS: Data of 242 HER2-positive breast cancer cases were obtained from the NAT dataset GSE243375 in the GEO database.Differentially expressed genes related to p CR were screened through differential expression analysis, LASSO regression,and support vector machine recursive feature elimination(SVM-RFE). Pathological tissue paraffin blocks and clinicopathological data were collected from 108 HER2-positive breast cancer patients who received NAT. The chi-square test was used to analyze correlations between expression of these two proteins and clinicopathological characteristics. The Mann-Whitney U test was employed to compare differences in protein expression levels between the p CR group and the non-pCR group. Spearman rank correlation analysis was performed to evaluate correlations between their expression and MP grade. Furthermore,binary logistic regression and ordinal logistic regression were used to adjust for confounding variables and to analyze independent predictive values of IGHG1 and TMSB10 for p CR and MP grade. RESULTS: Bioinformatics analysis identified 10 differentially expressed genes related to p CR after NAT in HER2-positive breast cancer. The AUC values for predicting p CR by IGHG1 and TMSB10 were0.626 and 0.618, respectively. Immunohistochemical results showed that TMSB10 was mainly expressed in the cytoplasm of breast cancer tumor cells,presenting brown-yellow granular staining. IGHG1 was mainly expressed in the cytoplasm of breast cancer tumor cells, showing diffuse or focal brown-yellow granular staining, with occasionally positive expression on the cell membrane. Univariate analysis revealed that expression levels of TMSB10 were significantly higher in the p CR group than in the non-pCR group(P=0.038),and the proportion of IGHG1 high expression in the p CR group(15.3%) was also significantly higher than that in the non-pCR group(2.0%)(P=0.043). Multivariate Logistic regression analysis showed that TMSB10 was an independent predictor of increased MP grade[OR=1.232,95%CI(1.060,1.431),P=0.006],and IGHG1 was also an independent predictor of increased MP grade[OR=1.434,95%CI(1.028,2.000),P=0.034]. The combined prediction AUC value for p CR by both indicators was 0.660 [95%CI(0.557,0.763)],which was superior to a single indicator. CONCLUSION: IGHG1 was shown to be an independent predictor of increased MP grade after neoadjuvant therapy in HER2-positive breast cancer,and its high expression was significantly associated with higher p CR rates. TMSB10 was an independent predictor of p CR and increased MP grade. IGHG1 and TMSB10 can be considered to be potential biomarkers for predicting efficacy of NAT in HER2-positive breast cancer. The combined application of IGHG1 and TMSB10 could improve accuracy of predicting the efficacy of NAT in HER2-positive breast cancer.