OBJECTIVE： To investigate effects of the co-stimulatory molecule B7-H4 on MFC cell proliferation and tumor formation， and to explore its potential mechanisms. METHODS： MFC cells were transfected with lentiviruses to knock down B7-H4 expression，and stable transfection cell lines were screened. The cells were divided into a knockdown group (transfected with B7-H4 knockdown lentivirus) and a control group (transfected with the empty vector lentivirus). Expression levels of B7-H4 mRNA and protein in MFC cells were detected using RT-qPCR and Western blot to evaluate knockdown effects. Impact of B7-H4 knockdown on MFC cell proliferation and migration was assessed through CCK-8 assay and cell scratch experiments. The stably transfected cells from both groups were then injected subcutaneously into mice， and tumor formation was evaluated through tumor transplantation experiments. Expression levels of B7-H4，WNT7A，and STAT3 in MFC cells and mouse tumor tissues were analyzed using RT-qPCR and Western blot. RESULTS： RT-qPCR and Western blot results showed that compared to the control group，expression levels of B7-H4 in the knockdown group were significantly reduced (P<0.01)，indicating successful construction of B7-H4 knockdown cell lines. In MFC cells， B7-H4 knockdown significantly reduced both proliferation and migration capacity compared to the control group (P<0.01). In subcutaneous tumor formation experiments， the tumor volume and mass in the knockdown group were significantly smaller than those in the control group (P<0.01). RT-qPCR and Western blot results demonstrated that expression levels of B7-H4， WNT7A， and p-STAT3 in the knockdown group were significantly lower than those in the control group (P<0.01). In contrast， total STAT3 expression showed no significant change (P>0.05). CONCLUSION： B7-H4 knockdown inhibited MFC cell proliferation and migration and affected tumor formation in MFC cells. Low expression of B7-H4 inhibited expression of WNT7A and STAT3.

OBJECTIVE： To investigate the role of autophagy in inducing ferroptosis in human colon cancer SW620 cells treated with 5-fluorouracil (5-FU) combined with oxaliplatin (L-OHP). METHODS： The CCK-8 assay was used to assess the effects of different concentrations of 5-FU and L-OHP acting alone on SW620 cell proliferation after 24 h. The IC₅₀ values for both drugs were calculated，and the IC₅₀ concentrations were selected for subsequent combination treatment. SW620 cells were then divided into eight groups： control， 5-FU，L-OHP，5-FU +L-OHP，autophagy inhibitor 3-methyladenine (3-MA)，3-MA+5-FU，3-MA+L-OHP， and the triple combination of 3-MA， 5-FU and L-OHP. Reagent kits were used to detect reactive oxygen species (ROS) and malondialdehyde (MDA) levels in each cell group. Western blot analysis was employed to assess the expression of ferroptosis-related proteins SLC7A11， GPX4， and ACSL4， as well as autophagy-related protein LC3B. Transmission electron microscopy was utilized to observe mitochondrial morphology. RESULTS： Compared with the control group， both 5-FU and L-OHP individually inhibited SW620 cell proliferation (P<0.01). The calculated IC₅₀ values for cell survival under single treatment with 5-FU and L-OHP were 2 971 and 185 μmol/L，respectively. Combined treatment at the IC₅₀ concentrations of both drugs further inhibited SW620 cell proliferation (P<0.01). Both single and combined treatments with 5-FU and L-OHP significantly increased ROS and MDA levels (P<0.01)，while decreasing the ferroptosis-related proteins SLC7A11 and GPX4 (P<0.01)， and increased ACSL4 expression (P<0.05). Following intervention with the autophagy inhibitor 3-MA， compared to the control group， the triple combination group exhibited decreased expression of ferroptosis-related proteins SLC7A11 and GPX4 (P<0.01) and increased ACSL4 expression (P<0.01). In the L-OHP-treated group and the 5-FU and L-OHP group，intracellular autophagy-related protein LC3B-II/I expression was upregulated (P<0.05 or P<0.01). Compared with the 5-FU and/or L-OHP groups，the triple-drug combination group exhibited further reduced cell viability (P<0.01) and significantly elevated MDA and ROS levels (P<0.01). In the 5-FU and/or L-OHP intervention groups supplemented with 3-MA，ACSL4 protein expression increased (P<0.05 or P<0.01)，while SLC7A11 and GPX4 proteins expression decreased (P<0.05 or P<0.01). Transmission electron microscopy revealed that 5-FU and/or L-OHP induced mitochondrial damage in SW620 cells，with more pronounced morphological alterations observed in the combination treatment group.CONCLUSION： 5-FU and L-OHP inhibited proliferation of SW620 colon cancer cells and induced ferroptosis. Co-treatment further enhanced the level of ferroptosis in cancer cells. Inhibition of autophagy potentiated the effect of combined 5-FU and L-OHP treatment.

OBJECTIVE： To evaluate relationships between inflammation-nutrition composite indexes and survival prognosis in patients with type I endometrial cancer (EC). METHODS： A total of 404 patients with type I EC diagnosed by postoperative pathology from January 2010 to May 2022 were included in this study. Complete blood count and biochemical examination results were collected within one week before surgery. Four inflammatory nutritional composite indicators were calculated： systemic inflammatory response index (SIRI)， aggregate index of systemic inflammation (AISI)， geriatric nutritional risk index (GNRI)， and advanced lung cancer inflammatory index (ALI). Clinicopathological data and follow-up data were also collected. The optimal cutoff value was determined using receiver operating characteristic (ROC) curves. Survival curves were plotted using the Kaplan-Meier method. Univariate and multivariate Cox proportional hazards regression models were used to screen for independent prognostic factors of overall survival (OS). RESULTS： The area under the curves for the inflammatory nutritional indicators were ranked as follows： ALI (0.609) > AISI (0.592) > SIRI (0.581) > GNRI (0.556). Univariate Cox regression showed that age ≥ 60 years，lymph node metastasis，FIGO stage (III + IV)，carbohydrate antigen 125 (CA125) ≥ 35 U/mL，and ALI<64.32 were significantly associated with shorter OS (all P<0.05). Multivariate Cox regression showed that advanced age [HR=2.25，95% CI(1.02， 4.96)，P=0.044]，high FIGO stage [HR=2.80，95% CI(1.27，6.20)，P=0.010]，and low ALI [HR=0.28，95% CI (0.08， 0.96)， P=0.042] were independent risk factors for OS， while SIRI， AISI， and GNRI were not significantly associated with OS (P>0.05). CONCLUSION： Among the four inflammatory and nutritional composite indicators，preoperative ALI had the best prognostic ability for type I EC，and ALI<64.32 was an independent risk factor for poor prognosis.

OBJECTIVE： To investigate the usefulness of thin-prep cytologic test (TCT) combined with high-risk human papilloma virus (HR-HPV) detection in opportunistic screening for cervical cancer in community postmenopausal women. METHODS： The clinical data of 698 community postmenopausal women who underwent opportunistic screening for cervical cancer from November 2021 to December 2024 were collected in this study， and these women were given TCT and HR-HPV detection. Among them， 165 individuals with abnormal screening results underwent colposcopy at a higher-level hospital. Finally， the sensitivity， specificity and accuracy of TCT， HR-HPV and their combined detection were compared and analyzed with colposcopy biopsy as the gold standard. The receiver operating characteristic (ROC) curve was used to analyze diagnostic performance. RESULTS： The sensitivities of TCT and HR-HPV testing were 91.11% and 92.22%，respectively，both with a specificity of 48.0%，positive predictive values of 67.77% and 68.03% ， negative predictive values of 81.82% and 83.72% ， and accuracies of 71.52% and 72.12% . The combined detection sensitivity of the two methods was 97.78% ， specificity was 45.33% ， positive predictive value was 68.22%，negative predictive value was 94.44%，and accuracy was 73.94%. The accuracy of TCT and HR-HPV testing alone was lower than that of the combined testing；the area under the ROC curve for combined testing in cervical cancer screening was higher than that of TCT or HPV alone (P<0.05). CONCLUSION： TCT combined with HR-HPV detection showed high diagnostic efficacy，improved sensitivity of cervical cancer screening， and higher clinical value for opportunistic cervical cancer screening in postmenopausal women.

OBJECTIVE： To investigate effects of saxitoxin (STX) exposure on cognitive function in F1 generation mice and its underlying mechanisms. METHODS： C57BL/6J mice were exposed to STX via drinking water during gestation and lactation to adulthood at doses of 0.00， 0.02， 0.20， or 2.00 μg/kg. Cognitive and memory functions were assessed in postnatal day 60 (PND60) and PND180 mice using the Morris water maze test， cortical neurotoxicity was evaluated by NeuN immunohistochemical staining， differentially expressed proteins in cortical tissues were screened and analyzed via Tandem Mass Tag (TMT) quantitative proteomics and enrichment analysis， key proteins were validated by Western blot， and cortical ATP levels were measured with a commercial kit. RESULTS： At STX doses of 0.02，0.20，and 2.00 μg/kg， no significant differences in maternal body weight，water intake，or body weight gain of F1 generation mice compared with the control group (P>0.05). In the Morris water maze， PND180 offspring exposed to STX exhibited prolonged platform-finding latency， reduced platform crossings， and decreased target quadrant occupancy compared to the control group (all P<0.05)， while no differences were detected at PND60. Immunohistochemical analysis revealed significantly fewer cortical neurons in PND180 mice following STX exposure (P<0.05). Proteomic profiling identified 295 DEPs at PND60 and 327 DEPs at PND180 in the 2.00 μg/kg group. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses highlighted oxidative phosphorylation (OXPHOS) as a critical pathway in both age groups. Western blot confirmed elevated expression of OXPHOS-related proteins Ndufs1 and Ndufb9 in PND180 cortical tissues (P<0.05)， with no significant changes at PND60. Cortical ATP levels were also significantly reduced in STX-exposed PND180 mice (P<0.05). CONCLUSION： STX exposure from gestation and lactation to adulthood induces cognitive impairment in F1 generation mice，and its mechanism of action may involve the oxidative phosphorylation pathway.

OBJECTIVE： To investigate the role of the mitochondrial glutathione (GSH) transporter SLC25A39 in regulating mitochondrial homeostasis and apoptosis in hepatoma cells. METHODS： Data on expression of SLC25A39 in tumor and adjacent tissues and its relationship with the survival time of hepatocellular carcinoma patients were obtained using the HCCDB public database. SLC25A39-knockdown HepG2 cells were verified by real-time quantitative PCR (qPCR) and Western blot. Impact of SLC25A39 expression on the proliferation of liver cancer cells was investigated using the CCK-8 method. HepG2 cells were treated with 50 μmol/L tert-butyl hydroperoxide (TBHP) for 12 hours to induce cell apoptosis. Cell viability was assessed using the CCK-8 assay，and cell apoptosis rates were detected by Annexin V/PI method. In addition，mitochondrial GSH content was detected by MitoRT staining and purified mitochondrial methods. Mitochondrial membrane potential was detected by JC-1 staining. Mitochondrial reactive oxygen species (ROS) levels were detected by MitoSOX staining. Intracellular ATP levels were detected by bioluminescence assay. RESULTS： Results from the HCCDB public database indicate that levels of SLC25A39 in most hepatocellular carcinomas were higher than those in their adjacent tissues (P<0.05). Expression of SLC25A39 was positively correlated with poor prognosis in cancer patients. Compared to the control cells， the proliferation rate of SLC25A39 knockdown cells gradually decreased， and the apoptosis ratio significantly increased (all P<0.05). Compared to the control cells，knockdown of SLC25A39 in HepG2 cells resulted in decreased mitochondrial GSH， mitochondrial membrane potential， and cellular ATP content， while mitochondrial ROS levels were increased (all P<0.05). After stimulation with TBHP， the apoptosis rate in SLC25A39 knockdown cells significantly increased compared to the control group， and both mitochondrial oxidative stress and functional impairment were further exacerbated (P<0.05). CONCLUSION： SLC25A39 was capable of promoting hepatocellular carcinoma cells to maintain mitochondrial homeostasis and redox balance by enhancing mitochondrial GSH uptake capacity， thereby conferring enhanced anti-apoptotic and survival capabilities. Targeted inhibition of SLC25A39 may be useful for the treatment of hepatocellular carcinoma.

OBJECTIVE： To investigate effects of ionizing radiation on the mRNA expression profile in exosomes from the AHH-1 human peripheral blood B lymphoblast cell line and to investigate usefulness of the profile as radiation biomarkers. METHODS： Using the AHH-1 cell line as a model， culture supernatants were collected from control cells and cells exposed to 2 Gy or 5 Gy of γ-rays at 24 h postirradiation. Exosomes were isolated by differential ultracentrifugation and characterized via nanoparticle tracking analysis for size distribution，transmission electron microscopy for morphology，and Western blotting for the surface markers CD63 and TSG101. Differentially expressed mRNAs were screened using expression microarray and subsequently subjected to GO and KEGG enrichment analyses. To further examine the dose- and time-dependent responses of candidate genes， AHH-1 cells were irradiated with 0， 1， 2， 4， or 6 Gy of γ-rays，and exosomes were collected at 24 h and 48 h postirradiation. Exosomal RNA was extracted，and the relative expression levels of selected mRNAs were validated by quantitative realtime PCR. RESULTS： Compared to the control group，a total of 197 differentially expressed mRNAs were identified in the exosomes from the 2 Gy irradiated cells，with 184 being upregulated and 13 downregulated. Following 5 Gy irradiation， 192 differentially expressed mRNAs were identified， comprising 170 upregulated and 22 downregulated transcripts. Bioinformatic analysis indicated that these altered mRNAs influenced cellular processes such as proliferation and apoptosis by participating in pathways including the TGF-β and ErbB signaling pathways. Real-time quantitative PCR analysis of the relative expression changes of candidate genes revealed that at 24 h after 0-6 Gy γ-rays irradiation， ZNFX1， DDB2， SPDYE2B， and CHD3 mRNA exhibited a significant upregulation trend. At 48 h after 4 Gy γ-rays irradiation， LENG8， FBXW7， AKAP1， and ZGRF1 mRNA were significantly upregulated. Notably，the expression levels of LENG8 and FBXW7 mRNA at 48 h after 4 Gy irradiation were approximately three times that of the control group. CONCLUSION： Ionizing radiation significantly altered mRNA expression profile in exosomes derived from AHH-1 cells. The differentially expressed genes identified (ZNFX1， DDB2， SPDYE2B， CHD3， LENG8， FBXW7， AKAP1， and ZGRF1) showed potential as biomarkers for ionizing radiation exposure.

OBJECTIVE： To investigate effects of aflatoxin B1 (AFB1) exposure on livers of mice from a metabolic perspective， thereby providing data support for elucidating mechanisms of AFB1-induced hepatotoxicity. METHODS： BALB/c mice were randomly divided into two groups. After a 7-day acclimatization period，the AFB1 group received 0.75 mg/kg AFB1 daily via oral gavage，while the control group was administered an equivalent volume of physiological saline. These mice were sacrificed after 30 days， and liver tissues were rapidly frozen and stored in liquid nitrogen. Metabolomic analysis of liver tissues was performed using liquid chromatography-mass spectrometry (LC-MS). Differential metabolites between the two groups were identified，and enrichment analysis of the involved metabolic pathways was conducted. RESULTS： After 30 days of AFB1 exposure，metabolite expression profiles in livers of mice were significantly altered. A total of 1 608 metabolites were detected，among which 32 were unique to the AFB1 group. Compared with the control group，201 differential metabolites were identified，of which 107 were significantly upregulated and 94 were significantly downregulated. Changes in metabolic profiles were associated with the regulation of 12 metabolic pathways： 11 related to metabolism and 1 related to genetic information processing. The upregulated metabolites were enriched in five pathways. The downregulated metabolites were enriched in seven pathways. Additionally， this study identified the GSSG/GSH ratio， oxododecanedioic acid， and 18-hydroxy eicosapentaenoic acid as potential biomarkers of AFB1-induced liver injury. CONCLUSION： AFB1 exposure significantly altered metabolic profiles of livers，with alterations attributed to pathways including metabolism and information processing. These findings corroborate traditional mechanisms of AFB1-induced liver injury and provide metabolomic-level data for understanding potential mechanisms underlying long-term，low-dose AFB1 exposure-induced hepatotoxicity in mice.

OBJECTIVE： To investigate teratogenic effect of γ-polyglutamic acid in SD rats. METHODS： Pregnant rats (n=80) were divided into negative control group，positive (cyclophosphamide) control group，and low，medium，and high dose (750，1 500，3 000 mg/kg，respectively) γ-polyglutamic acid groups according to the National Food Safety Standard GB 15193.14-2015 and to their body weight in a zigzag pattern. Rats were orally administered with γ-polyglutamic acid at the designed dose per day from day 6 to day 15 after conception，while the positive control group was given a one-time intraperitoneal injection of 15 mg/kg cyclophosphamide on day 12 of conception. On the 20th day of pregnancy，they were euthanized，and their parental pregnancy and fetal development were examined by caesarean section. Uteri connected to fetuses were removed and weighed， the number of corpora lutea were recorded as well as premature deaths， late stillbirths，live births，and implantation. Fetel rats were inspected for the appearance，internal organs，and skeletal deformities. RESULTS： There was no statistically significant difference (P>0.05) in the uterine mass， corpus luteum number， implantation number， absorbed fetus， stillbirth， live fetus number， as well as the appearance，visceral，and skeletal deformities of the fetal mice between the γ-polyglutamic acid dose groups and the negative control group. However，there was a statistically significant difference (P<0.05 or P<0.01) in all indicators between the cyclophosphamide (15 mg/kg) group and the negative control group. CONCLUSION： Under the conditions of this experiment，γ-polyglutamic acid had no significant adverse effects on the body mass of pregnant rats，reproductive ability of rats，and growth and development of fetal rats. No teratogenic effects of γ-polyglutamic acid on the appearance，bones，and internal organs of fetal mice were observed.

OBJECTIVE： To evaluate safety of recombinant humanized Type III collagen and to provide reference data for safe clinical use. METHODS： The maximum tolerated dose method was adopted in the acute toxicity test. ICR mice aged 3-5 weeks were administered the recombinant collagen via tail vein injection at the maximum dosage of 2 000 mg/kg for a single administration. After administration，the toxic reaction in these mice was closely observed. In the long-term toxicity test，120 SD rats aged 6 weeks were selected and divided into high-dose (130 mg/kg)， medium-dose (65 mg/kg)， low-dose (32.5 mg/kg) groups and a control group，with 30 rats in each group. The drug was continuously administered for 28 days，followed by a 28-day recovery period. The main inspection indicators included animal ophthalmology， body weight， feed consumption，hematological indexes，serum biochemistry，blood coagulation，urine indexes，histopathology，etc. RESULTS： In the acute toxicity test，the maximum tolerated dose was greater than 2 000 mg/kg，equivalent to 1 503.76 times the clinically proposed dosage， and no obvious toxic and side effects. In the 28-day repeated dose toxicity test，the no-observed-adverse-effect level (NOAEL) of recombinant humanized Type III collagen administered via tail vein injection in SD rats was 130 mg/kg， corresponding to 97.74 times the proposed clinical dose for adults. CONCLUSION： Under the conditions of this experiment，recombinant type III humanized collagen did not show obvious toxic reactions.

OBJECTIVE： To evaluate safety of lactose-N-tetrasaccharide (LNT) according to national food safety standards. METHODS： Acute oral toxicity test was performed using the limit method. Twenty ICR mice were administered 20 g/kg of LNT via gavage once，and the toxic effects were observed for two consecutive weeks. Bacterial reverse mutation test： Five histidine-deficient Salmonella Typhimurium strains (TA97a，TA98， TA100， TA102， and TA1535) were used， and two tests were conducted with and without S₉. For the mammalian erythrocyte micronucleus test， ICR mice were administered LNT twice via gavage at a maximum dose of 5.0 g/kg，with medium and low doses of 2.5 and 1.25 g/kg，respectively，over 30 hours. A solvent control group and a positive control group were also included. For the in vitro chromosomal aberration test， Chinese hamster lung (CHL) cell line was used，and three concentrations (0.8，2.0，and 5.0 mg/mL) were set up with and without S₉. A DMEM negative control and a positive control group were also included. A 90-day oral toxicity test was conducted with three dosage groups (2.5，5.0，and 10.0 g/kg) and a solvent control group to observe sub-chronic toxicity in SD rats. RESULTS： The acute oral toxicity test showed that the LD₅₀ of LNT in mice was greater than 20 g/kg; all three mutagenicity tests were negative. In the 90-day oral toxicity test， no significant abnormalities were observed in female rats， with a no-adverse-effect level (NOAEL) of 10.0 g/kg. Male rats showed weight gain and decreased food intake， with a NOAEL of 5.0 g/kg. CONCLUSION： Continuous gavage administration of LNT for 90 days had no adverse effects on female rats， but it caused weight gain and decreased food intake in male rats. Therefore， the safety of LNT in food applications needs to consider gender differences.

OBJECTIVE： To study synergistic anti-fatigue effect of different doses and combinations of American ginseng and astragalus，and to screen out the dose with significant anti-fatigue effect. METHODS： American ginseng and astragalus were extracted with water and precipitated with alcohol at a ratio of 1∶3， and the precipitate was dried to obtain the herbal extract. The extract was administered to mice by gavage at high，medium，and low doses (600，300，and 150 mg/kg，respectively) for 30 days. A blank control group and a positive control group were also set up. The fatigue resistance of animals in different dose groups was judged by lactate clearance rate test， serum urea nitrogen and creatine kinase content after exercise， liver glycogen content，and changes in related indicators in the weighted swimming test. RESULTS： Compared with the blank control group，each test group showed a reduced area under the curve of serum lactate，increased liver glycogen content， reduced serum urea nitrogen and creatine kinase levels after exercise and increased time to exhaustion in the weighted swimming test (P<0.05 or P<0.01). Compared with the positive control group， the area under the curve (AUC) of blood lactate and the liver glycogen content of mice in the high-dose test substance group were decreased (P<0.01). CONCLUSION： The American ginseng and astragalus combination had an anti-fatigue effect on mice，and the high-dose group has a better anti-fatigue effect than the positive control.