OBJECTIVE： To investigate the effects of early-life Bisphenol A (BPA) exposure on glucose and lipid metabolism in post-weaning mice fed a high-fat diet. METHODS： Pregnant mice were exposed to BPA through drinking water at a concentration of 0.5 mg/(kg·d) from gestational day 6 until weaning of offspring. Twenty pregnant mice were divided equally into control and BPA groups. The weaned offspring were organized into，four groups：normal diet control，normal diet BPA，high-fat diet control，and high-fat diet BPA，with 10 mice per group，balanced by sex. Mice were sacrificed at postnatal day 21 (PND21) and postnatal day 91 (PND91) for the collection of blood and pancreas samples. Before sacrificing the PND91 mice，they were tested for glucose and insulin tolerance. Non-targeted lipidomics of the pancreas was analyzed using high-resolution liquid chromatography-mass spectrometry. qPCR was employed to measure mRNA expression levels of ceramide synthesis genes in the pancreas. Plasma insulin and pancreatic ceramide synthase activity were assessed using ELISA kits. Malondialdehyde (MDA) and superoxide dismutase (SOD) activities in the pancreas were measured using specific assay kits. RESULTS： At PND21，both male and female offspring in the BPA exposure group exhibited accelerated weight gain compared to the control group. At PND91，male mice in the high-fat diet BPA group showed increased body weight compared to the high-fat diet control group (P<0.05). Results of glucose tolerance and insulin tolerance tests at PND91 indicated that，compared to the high-fat diet control group，both male and female mice in the high-fat diet BPA group had significantly increased areas under the blood glucose curve (P<0.05). Calculating the insulin resistance index in mice，it was found that，male mice in the PND91 high-fat diet BPA group had an elevated insulin resistance index (P<0.05). Non-targeted lipidomics of the pancreas revealed that at PND21，compared to the control group，male and female mice in the BPA group had significantly increased ceramide (Cer) abundance (P<0.05). At PND91，compared to the high-fat diet control group，male mice in the high-fat diet BPA group had significantly increased ceramide abundance (P<0.05). Results from pancreatic qPCR showed that at PND21，compared to the control group，male mice in the BPA group had upregulated mRNA expression of the Cers4 and Cers6 genes，while female mice had upregulated Cers4 gene expression (P<0.05). At PND91，compared to the high-fat diet control group，male mice in the high-fat diet BPA group had upregulated mRNA expression of the Cers2，Cers4，and Cers6 genes (P<0.05). Analysis of ceramide synthase activity in the pancreas indicated that at PND21，compared to the control group，male mice in the BPA group had significantly enhanced Cers2，Cers4，and Cers6 activity (P<0.05). At PND91，compared to the high-fat diet control group，male mice in the high-fat diet BPA group had significantly enhanced Cers2 and Cers4 activity (P<0.05). Assessment of oxidative and antioxidative indicators showed that at PND91，compared to the high-fat diet control group，male mice in the high-fat diet BPA group had significantly increased pancreatic MDA content and significantly decreased SOD activity (P<0.05). Correlation analysis indicated a significantly positive correlation between ceramide abundance and the oxidative/antioxidative ratio in the pancreas of PND91 male mice (P<0.05). CONCLUSION： Early-life BPA exposure combined with post-weaning high-fat diet impaired glucose tolerance in offspring mice. It increased the de novo synthesis of pancreatic ceramide in male offspring mice in a sex-specific manner，which was closely related to the imbalance of oxidative and antioxidative status in the pancreas of male mice.

OBJECTIVE： This study aimed to develop a method for detecting white blood cell subsets with high telomerase reverse transcriptase (TERT) activity to evaluate proliferative potential of white blood cells in patients with malignant tumors. METHODS： A modified recombinant type I herpes simplex virus (HSV1-hTERTp-eGFP) was able to replicate in white blood cells with high TERT promoter transcriptional activity and to express green fluorescent protein (GFP). By observing green fluorescence，TERT positive white blood cells with high proliferation potential was identified，and the immune proliferation potential of individuals was evaluated. The addition of 100，500 and 1 000 GFP positive Jurkat cells that were infected with HSV1-hTERTp-eGFP virus for 24 hours to 2.5×10⁶ peripheral blood white blood cells that have not been infected with the virus were used as simulated samples for detection. Three parallel groups were set up and were tested for recovery rate，repeatability，linear regression coefficient，and other indicators. At the same time，peripheral blood samples were collected from 5 patients with benign pulmonary nodules，and 3 parallel groups were set up to detect the proportion of TERT positive white blood cells in 1 mL of peripheral blood in each group. The technical reliability of detecting peripheral blood samples was evaluated by calculating the coefficient of variation within each group. Using this detection technology，the percentage of TERT positive white blood cells in peripheral blood of 80 patients with benign pulmonary nodules and 80 patients with primary lung adenocarcinoma were detected and analyzed. RESULTS： In the simulated samples with Jurkat cells，the average number of positive cells were 80.3，448.2 and 920.6，respectively，with recovery rates of 80.3%，89.6% and 92.1%，respectively；The inter batch coefficients of variation for the three repeated experiments were 11.3%，7.2% and 4.4%，respectively，while the intra batch coefficients of variation within the three parallel experimental groups were less than 10%；The intra group coefficient of variation for the percentage of TERT positive cells in peripheral blood of 5 patients with benign pulmonary nodules was also within 10%. This detection technology showed high accuracy and stability. In 80 patients with benign pulmonary nodules，the percentage of TERT positive white blood cells showed a decreasing trend with age (r=-0.61；P<0.01). Compared with the control group of benign pulmonary nodule patients，the percentage of TERT positive white blood cells in early lung adenocarcinoma patients decreased (P<0.01)；Compared with non-invasive lung adenocarcinoma patients，the proportion of TERT positive white blood cells in invasive lung adenocarcinoma patients was significantly reduced (P<0.01). CONCLUSION： The peripheral blood immune cells of patients with lung adenocarcinoma showed a decrease in proliferation ability，which was more pronounced in patients with invasive lung adenocarcinoma. The observation was reminiscent to immunosenescence，and would provide a better understanding of poor prognosis among patients with invasive lung adenocarcinoma.

OBJECTIVE： To explore the role and mechanism of the miR-520c-3p/CXCL8 axis in erythroid differentiation of chronic myeloid leukemia (CML). METHODS： K562 cells were induced to differentiate with hemin and treated with CXCL8 (0，20，40，60，80 and 100 ng/mL) for 72 h. Expression of γ-globin mRNA was examined by RT-qPCR to analyze the effect of CXCL8 on erythroid differentiation. miR-520c-3p mimics and inhibitor were transfected into K562 cells to enhance or inhibit miR-520c-3p function，respectively. Expression of γ-globin mRNA was measured by RT-qPCR to assess the impact of miR-520c-3p on erythroid differentiation of K562 cells. Western blot and RT-qPCR experiments were performed to detect CXCL8 expression and observe the influence of miR-520c-3p on CXCL8 expression. The cells were divided into the experiment group (co-transfected with miR-520c-3p mimics and CXCL8 3'-UTR) and the control group (co-transfected with mimics-control and CXCL8 3'-UTR)，a dual luciferase reporter gene assay was conducted to examine the targeting relationship between miR-520c-3p and the 3'-UTR of CXCL8. RESULTS： Compared to the control group，expression of CXCL8 mRNA was significantly increased after hemin-induced differentiation (P<0.01). Compared to the untreated group，the γ-globin mRNA expression of CXCL8 treatment groups were significantly elevated (P<0.01). Compared with the negative control group，the expression levels of γ-globin mRNA，CXCL8 mRNA，and CXCL8 protein in K562 cells transfected with miR-520c-3p mimics were all decreased (P<0.01). In cells transfected with the miR-520c-3p inhibitor，expression levels of γ-globin mRNA，CXCL8 mRNA，and CXCL8 protein were all increased (P<0.01). The dual luciferase reporter gene assay revealed a significant decrease in relative fluorescence activities in the experimental group compared to the control group (P<0.01). CONCLUSION： This study reveals the key role of the miR-520c-3p/CXCL8 signaling axis in the erythroid differentiation of CML cells. miR-520c-3p exerted an inhibitory effect on the erythroid differentiation of K562 cells by targeting the expression of CXCL8.

OBJECTIVE： To investigate the role of the Wnt1-Ngn2-En1/2 signaling pathway in the herbicide simazine-induced damage to the dopaminergic neurons in embryonic and weaned mice. METHODS： Fifteen female and fifteen male healthy SPF Kunming mice were kept in containers in a 2∶1 ratio. During pregnancy，three groups of maternal mice were randomized∶control group (0.3% methyl cellulose solution)，low-dose group (50 μg/kg) and high-dose group (200 μg/kg). The tissues of the offspring on the day 8.5 (E8.5)，day 12.5 (E12.5)，and postnatal day 21 (PND21) of development were obtained. mRNA and protein expression of the Wnt1-Ngn2-En1/2 signaling pathway-related genes (Wnt1，Ngn2，En1，En2，Nr4a2 and Th) were detected using immunofluorescence hybridization (FISH)，real-time fluorescence quantitative PCR (qPCR) and Western blot. Through an open field experiment，effect of the treated pregnant mice on the autonomic behavior of PND21 female and male mice were identified. RESULTS： Western blot and qPCR data in E8.5 demonstrated that in comparison to the control group，low simazine-exposed groups had a decrease in the mRNA and protein expression of Wnt1 and Ngn2 (P<0.05)，and the high-dose group had a substantial decline in the expressions of Wnt1，Ngn2，En1 and En2 (P<0.05). The results of E12.5 embryo show the mRNA and protein expression of Nr4a2 and Th in the low-dose group declined as compared with the control group，(P<0.05)，and the expression of Nr4a2，Th，Wnt1，Ngn2，En1 and En2 in high-dose group decreased significantly compared with that in control group (P<0.05). The FISH data verified that Wnt1 mRNA expression was substantially reduced in the low-dose and high-dose sections of E8.5 and E12.5 as opposed to that of the control group (P<0.05). The high-dose group of PND21 offspring exhibited significantly reduced motor and self-exploration abilities in comparison to the male offspring of the control group (P<0.05)，and there was a notable down-regulation of Wnt1 and Th protein expressions (P<0.05). CONCLUSION： Simazine affected the development of dopaminergic neurons in male offspring through the Wnt1-Ngn2-En1/2 signaling pathway，and damaged the ability of independent behavior of male mice.

OBJECTIVE： To investigate the involvement of Dickkopf-related protein-1 (DKK-1) and nicotinamide N-methyltransferase (NNMT) in clinicopathology and prognosis among liver cancer patients. METHODS： Expression levels of NNMT and DKK-1 mRNA in normal liver tissues and liver cancer cells were analyzed by GEPIA2 database.，Effects of the two on the overall survival (OS) and disease-free survival (DFS) of patients with liver cancer were analyzed. Immunohistochemistry was used to detect the protein expression of NNMT and DKK-1 in liver tumor group，benign lesion group and normal group，and the relationship between them and the occurrence and development of liver cancer and clinicopathological features was analyzed. RESULTS： The GEPIA2 database showed that expression levels of NNMT mRNA in liver cancer tissues were significantly lower than that in normal liver tissues (P<0.05). Expressions of NNMT and DKK-1 mRNA in liver cancers were negatively correlated (r=-0.13，P<0.05). Both had no significant effect on the 10-year OS and DFS of liver cancer patients. The 5-year survival rate of liver cancer patients with high expression of NNMT mRNA was higher than that of those with low expression (P<0.05). The results of immunohistochemistry showed that compared with normal liver tissues，the expressions of DKK-1 and NNMT proteins in liver cancer tissues were significantly reduced (P<0.01). Expression of DKK-1 protein in hepatocellular carcinoma was significantly higher than that in cholangiocarcinoma. The expression of both proteins was not significantly correlated with tumor size，stage and tissue grade (P>0.05). CONCLUSION： The expression levels of DKK-1 and NNMT proteins in liver cancer tissues are lower than those in normal liver tissues. The expression levels of DKK-1 protein were related to the tissue types of liver cancers，and the expression levels of NNMT mRNA affected the 5-year survival rate of patients with liver cancer.

OBJECTIVE： To study the proportion of HER-2-low expression in breast cancer and the clinicopathological characteristics of HER-2-low breast cancer. METHODS： From January 2019 to December 2023，982 cases of invasive breast cancer cases that were treated in the Shenzhen Maternal and Child Healthcare Hospital were collected. Immunohistochemistry (IHC) was used to detect the expression of human epidermal growth factor receptor-2 (HER-2)，hormone receptors including estrogen receptor (ER) and progesterone receptor (PR)，and cell proliferation related protein Ki-67. Fluorescence in situ hybridization (FISH) was used to detect HER-2 gene amplification. The clinicopathological data such as the patients-- age，tumor diameter，histological grades of tumor and lymph node metastasis status were retrospectively studied to analyze the proportion of HER-2 low expression in breast cancer cases and the difference in clinicopathological characteristics between HER-2-low breast cancer and HER-2 negative/positive breast cancer. RESULTS： Among the 982 cases，567 (57.74%) were HER-2-low，194 (19.76%) were HER-2 negative，and 221 (22.50%) were HER-2 positive. The histological grade and Ki-67 proliferation index of HER-2-low breast cancer were lower than those of HER-2 negative and HER-2 positive breast cancer (all P<0.01). The lymph node metastasis rate in HER-2-low breast cancer was higher than it in HER-2 negative breast cancer，but lower than it in HER-2 positive breast cancer (all P<0.01). The positive rate of hormone receptors in HER-2-low breast cancer was higher than that in HER-2 positive breast cancer (P<0.01)，but no significant difference with HER-2 negative breast cancer. There was no significant difference between HER-2-low breast cancer and HER-2 negative or HER-2 positive breast cancer in terms of patients-- age and tumor diameter. CONCLUSION： Breast cancer with low HER-2 expression accounted for a high proportion of the overall breast cancer cases，and showed low histological grade，low Ki-67 proliferation index，but high hormone receptor positivity rate. These characteristics may affect therapeutic efficacy.

OBJECTIVE： To evaluate the value of serum cystatin 4 (CST4) levels in the clinical diagnosis and prognosis evaluation of pancreatic cancer. METHODS： From October 2020 to January 2023，71 newly diagnosed pancreatic cancer patients who were treated in the Fourth Hospital of Hebei Medical University were selected for our study，and their clinical indicators including blood routine，coagulation function，blood biochemistry，and serum tumor markers were collected. Differences in CST4 and other clinical laboratory test results between these patients and patients with benign pancreatic tumors were analyzed. Receiver operating curve (ROC) analysis were performed to evaluate the significance of CST4 in the diagnosis and clinical progression assessment of pancreatic cancer. Kaplan-Meier survival curves were constructed，and multivariate Cox regression analysis was conducted to evaluate the value of CST4 in the clinical prognosis assessment of pancreatic cancer compared with traditional tumor markers：carcinoembryonic antigen (CEA) and carbohydrate antigen 199 (CA19-9). RESULTS： Compared with the benign tumors group，the proportions and median values of serum CST4，CA19-9，and CEA levels in pancreatic cancer patients were significantly increased (P<0.01). ROC analysis showed that the area under the curve (AUC) of CST4 in the diagnosis of pancreatic cancer was 0.782，and the AUC of CST4 combined with CEA and CA19-9 was significantly higher than that of single detection (AUC=0.930) (P<0.05). CST4 showed higher value in predicting the clinical progression of pancreatic cancer (AUC=0.794)，with a sensitivity of 80.90%，specificity of 70.80%，positive likelihood ratio of 2.77，and an optimal cutoff value of 84.41 U/mL. Prognostic analysis showed that the median survival times of high CST4 group and low CST4 group patients were significantly different at 6.2 months and 25.6 months (P<0.01). Elevated serum CST4 level was identified as an independent risk factor for the prognosis of pancreatic cancer (P<0.05). CONCLUSION： Our data show that serum CST4 level can be used as a clinical auxiliary diagnostic indicator for pancreatic cancer and as an independent risk factor for the prognosis of pancreatic cancer.

OBJECTIVE： This study aimed to elucidate the distribution，accumulation and toxicity of cadmium in the blood，esophagus，liver，and kidneys of mice following subchronic exposure of cadmium in drinking water. METHODS： For this study，27 pathogen-free (SPF)-grade C57BL/6J mice were randomly assigned to a control，low-dose (10 mg/kg CdCl₂)，and high-dose groups (75 mg/kg CdCl₂). The control group received standard feed and double-distilled water，while the exposed groups were given standard feed and a solution containing the corresponding concentration of CdCl₂. The low-dose group was exposed to cadmium for 8，12，16，and 20 weeks，and the high-dose group for 16 weeks，with three mice per time point. Whole blood was collected from the orbital cavity of mice of the control group at 8 and 12 weeks，the low-dose group at 8，16，and 20 weeks，and the high-dose group at 16 weeks. Mice were sacrificed according to the experimental time points，and esophageal，liver，and kidney tissues were collected. Cadmium levels were measured using inductively coupled plasma mass spectrometry (ICP-MS). RESULTS： Blood cadmium levels in the treated groups were significantly higher than in the control group (P<0.01). The blood cadmium levels in the low-dose group were significantly increased compared to the control (P<0.01)，but there was no statistically significant difference among the different exposure time points within the group (P>0.05). At the 16-week time point，the blood cadmium levels in the high-dose group were significantly higher than in the low-dose group (P<0.01). The esophageal cadmium concentration in the low-dose group was higher than in the control group at all exposure time points (P<0.05 or P<0.01)，with the highest concentration at 20 weeks (67.7 μg/L)，which was statistically significant compared to 8 weeks (44.0 μg/L) and 12 weeks (35.2 μg/L) (P<0.05 or P<0.01). The esophageal cadmium concentration in the high-dose group (129.2 μg/L) at 16 weeks was significantly higher than in the low-dose group (47.9 μg/L，P<0.01). In the low-dose group，cadmium concentrations in liver and kidney tissues increased from 12-16 weeks and 8-16 weeks，respectively，compared to the control group (P<0.05 or P<0.01)，reaching a peak at 16 weeks (P<0.01) before declining. At 16 weeks，there were statistically significant differences in the cadmium concentrations of liver and kidney tissues among all dose groups，with the highest concentrations in the high-dose group (P<0.01). Cadmium levels were quantifiable in the blood，esophagus，liver，and kidneys of both control and cadmium-treated groups，with the highest to lowest order of cadmium content being kidney>liver>esophagus>blood. CONCLUSION： Dietary cadmium exposure resulted in the accumulation of cadmium in the blood，esophagus，liver，and kidney of mice，exhibiting a definite time- and dose-dependent effect.

OBJECTIVE： To investigate involvement of serum selenium and urine iodine concentrations (UIC) with thyroid cancer through meta-analysis. METHODS： Systematic search was conducted through PubMed，Web of Science，Wanfang Database and CNKI for relevant publications up to November 2023. Meta-analyses were performed using Review Manager 5.4.1 and Stata 17. RESULTS： A total of 18 reports including 22 case-control studies with 6 457 participants were included. Pooled analysis showed that the serum selenium level of thyroid cancer patients was lower than that of healthy controls [SMD=-1.46，95%CI=(-2.21，-0.72)，P=0.001]. Compared with the healthy control group，excessive iodine intake (UIC≥300 μg/L) was positively correlated with the occurrence of thyroid cancer [OR=2.68，95% CI=(1.40，5.12)，P=0.003]，and the appropriate iodine intake (100≤UIC<200 μg/L) was negatively correlated with the occurrence of thyroid cancer [OR=0.47，95%CI=(0.29，0.76)，P=0.002]，and insufficient iodine intake (UIC<100 μg/L) [OR=0.62，95%CI=(0.35，1.09)，P=0.10] and minor iodine overdose (200≤UIC<300 μg/L) [OR=0.81，95%CI=(0.50，1.32)，P=0.40] were not statistically significant with the occurrence of thyroid cancer. CONCLUSION： This meta-analysis indicates that decreased serum selenium levels may elevate the risk of developing thyroid cancer，whereas excessive iodine intake could serve as a potential risk factor for thyroid cancer. Conversely，moderate iodine intake may confer protection against thyroid cancer.

OBJECTIVE： To investigate the characteristics and mechanisms of cisplatin-induced polyploid giant cancer cells (PGCCs) in vitro. METHODS： Cultured cells were treated with cisplatin at concentrations of 1.5，3，6，12，and 24 μg/mL for 3，4，and 5 days. DNA content was analyzed by flow cytometry to determine the optimal combination of concentration and duration for PGCC induction in A549，HepG2，and SK-OV-3 cells. Morphological changes and DNA nuclear area were examined through morphological observation and Giemsa staining. Expression of stemness genes (Nanog，Sox-2，OCT-4，c-Myc) was evaluated by RT-qPCR，while stemness surface markers (CD44，CD133) were analyzed by flow cytometry. Cell senescence was assessed by β-galactosidase staining. After 72-hour treatment with cisplatin at concentrations ranging from 0.75 to 96 μg/mL，cell viability was measured by the MTT assay. RESULTS： Treatment with 3 μg/mL cisplatin for 3 d effectively induced polyploidy (>4N) in all three cell lines. The induced cells exhibited significantly increased cell and nuclear areas；varying degrees of upregulation in stemness markers CD44 and CD133，along with partial increase in stemness gene expression；partial senescence phenotype in A549 cells；and enhanced cisplatin tolerance in A549 and SK-OV-3 PGCCs compared to controls. CONCLUSION： The cisplatin-induced PGCC model in A549 cells demonstrated enlarged cell and nuclear areas，expressed stemness genes (Nanog，Sox-2，OCT-4)，and showed increased cisplatin tolerance. The model may be useful for investigating，drug resistance mechanisms.

OBJECTIVE： To evaluation of teratogenicity and mutagenicity of refined fish oil (DHA content 89.97%，EPA content 1.12%). METHODS： SD female rats were given different doses(1.2，2.3，4.6 g/kg) of refined fish oil by oral administration on the 6th to 15th day of pregnancy. The body weight，pregnancy outcome，fetal viscera and bone，and fetal growth and development of pregnant rats were measured to evaluate the teratogenic effect of the fish oil. Effects of refined fish oil on the colony number and colony morphology of Salmonella typhimurium TA97a，TA98，TA100，TA102 and TA1535 were detected under activated and non-activated conditions. Its mutagenicity was evaluated by detecting the micronucleus rate in bone marrow cells and chromosome aberration rate in spermatogonia from normal ICR mice. RESULTS： In the teratogenic test，the NOAEL of refined fish oil on pregnant mice was 4.6 g/kg. The Ames test results for all dose groups (8-5 000 μg/dish) was negative under both metabolic and non-metabolic activation conditions. Compared with the control group，the micronucleus rate，chromosome aberration rate and aberration rate in bone marrow cells for each dose group were insignificant (P>0.05)，and there was no obvious mutagenicity. CONCLUSION： Under the experimental conditions，no teratogenic and mutagenic effects were observed in refined fish oil.