[摘 要] 目的：探讨X连锁锌指蛋白（ZFX）通过Nectin-4表达影响食管鳞状细胞癌（ESCC）进展的分子机制及其对PI3K/AKT信号通路的激活作用。方法：收集2022年8月至2023年7月在南充市中心医院手术切除的30对ESCC组织及癌旁组织标本。采用人食管上皮细胞HET-1A及ESCC细胞KYSE-30、KYSE-150、KYSE-410、KYSE-510和TE-1。基于2018年至2019年收集的6例ESCC配对标本转录组测序数据筛选Nectin-4，通过TIMER2.0数据库、RT-qPCR和WB法、免疫组织化学（IHC）检测ESCC组织及细胞中Nectin-4的表达水平。采用shRNA技术在KYSE-410和KYSE-510细胞中敲低Nectin-4，通过CCK-8、克隆形成、划痕愈合及Transwell实验检测敲低Nectin-4对细胞增殖、迁移和侵袭能力的影响，采用WB法检测敲低Nectin-4对细胞PI3K/AKT通路及EMT相关蛋白表达水平的影响。通过生物信息学预测并结合双萤光素酶报告基因实验，鉴定并验证ZFX作为Nectin-4的上游转录调控因子。结果：综合分析显示，ESCC组织及细胞系中Nectin-4表达显著高于癌旁组织及HET-1A细胞（均P < 0.01）。功能研究表明，敲低Nectin-4显著抑制KYSE-410和KYSE-510细胞的增殖活性、克隆形成能力，以及迁移和侵袭能力（均P < 0.01）。机制方面，敲低Nectin-4时细胞中E-cadherin表达上调、N-cadherin下调（均P < 0.01），并抑制PI3K/AKT通路相关蛋白的磷酸化水平（P < 0.01）。结论： ZFX通过上调Nectin-4表达激活PI3K/AKT信号通路促进ESCC进展，为ESCC的治疗提供了潜在的新靶点。

Objective To investigate the mechanism by which hypoxia-inducible factor(HIF)-1α and circ-UBE2G1 mutually regulate and promote thyroid cancer(THCA)metastasis under hypoxic microenvironment.Methods The GEPIA database was used to analyze the expression characteristics and correlation between circ-UBE2G1 and HIF-1α in THCA.The relationship of circ-UBE2G1 and miR-330-3p with survival rate was analyzed using Kaplan-Meier survival curve.After THCA cells were exposed to hypoxia,HIF-1α was silenced by transfection to analyze its regulation for circ-UBE2G1 transcription.The targeting relationship between miR-330-3p and either circ-UBE2G1 or HIF-1α was verified by sequence prediction and dual luciferase reporter assay.The effects of HIF-1α overexpression and circ-UBE2G1 silencing on THCA cell migration and invasion were analyzed after corresponding transfections.A tumor-bearing nude mouse model was established by subcutaneous injection of THCA cells with HIF-1α overexpression and circ-UBE2G1 silencing,respectively.THCA tissues and adjacent normal samples were clinically collected to analyze the expression levels and correlations of circ-UBE2G1,miR-330-3p,and HIF-1α.Results Bioinformatics analysis showed that circ-UBE2G1 was positively correlated with HIF-1α(P<0.01),and its high expression was associated with a low survival rate in THCA patients(P=0.024).Inhibition of HIF-1α blocked the promotive effect of hypoxia on circ-UBE2G1(P<0.05).Silencing circ-UBE2G1 inhibited the migration and invasion of THCA cells(P<0.05),and reversed the promotive effect of HIF-1α on these processes(P<0.05).Dual luciferase reporter gene assay revealed that miR-330-3p targeted both circ-UBE2G1 and HIF-1α(P<0.05).Silencing circ-UBE2G1 led to increased levels of miR-330-3p(P<0.05),whereas increasing miR-330-3p inhibited the expression of HIF-1α(P<0.05).In clinical THCA samples,both circ-UBE2G1 and HIF-1α were increased and positively correlated(P<0.05),while miR-330-3p was lowly expressed and negatively correlated with both circ-UBE2G1 and HIF-1α(P<0.05).Conclusion Hypoxia promotes the transcription of circ-UBE2G1 by inducing HIF-1α expression,and circ-UBE2G1 promotes HIF-1α expression by targeting miR-330-3p,thereby promoting THCA metastasis.

Respiratory syncytial virus is the leading pathogen of lower respiratory tract infections in children under 5 years of age. Early-life respiratory syncytial virus infection is closely associated with long-term adverse outcomes，including impaired lung function，recurrent wheezing，and asthma. Metabolomics，an emerging systems biology approach，enables the quantitative analysis of dynamic changes in small-molecule metabolites within biological samples，providing critical insights into disease pathogenesis. This article reviews the metabolomic profiles of children with respiratory syncytial virus infection，with a focus on alterations in carbohydrate，lipid，and amino acid metabolism pathways. Additionally，it examines the distinct metabolomic features of children who develop recurrent wheezing following respiratory syncytial virus infection. These findings offer novel perspectives for elucidating the pathophysiological mechanisms of respiratory syncytial virus infection and improving early diagnosis and prognostic assessment.

To clarify the occurrence and distribution of broad bean wilt virus 2(BBWV2) on Rehmannia glutinosa, this study collected 87 R. glutinosa samples with typical symptoms of viral disease such as chlorosis and crumple from Wenxian county and Wuzhi county in Jiaozuo city, Henan province and Qiaocheng district in Bozhou city, Anhui province. The BBWV2 CP target band was amplified from 37 R. glutinosa samples by RT-PCR technology. The total detection rate reached 42.5%, among which 43.0% was detected in samples from Henan province. The detection rate in samples from Anhui province was 37.5%. 37 BBWV2 CP sequences were obtained by cloning and sequencing of BBWV2 positive samples(data has been submitted to GenBank, accession numbers: PP407959-PP407995), and the sequence analysis of these CP sequences with 91 other BBWV2 isolates in GenBank showed a high genetic diversity with a consistency rate of 70.8%-100%. Meanwhile, phylogenetic analysis showed that BBWV2 could be divided into three groups according to CP sequences, among which the BBWV2 in R. glutinosa isolates obtained in this study were all located in group 3. This study identified the differences in the occurrence, distribution, and genetic diversity of BBWV2 in R. glutinosa from Henan province and Anhui province and provided a theoretical basis for the prevention and control of BBWV2.
Rehmannia/virology* ; Phylogeny ; Plant Diseases/virology* ; China ; Molecular Sequence Data ; Fabavirus/classification*

Objective:To investigate the value of electrical cardiometry(EC),a noninvasive hemodynamic testing technique,in the di-agnosis of heart failure(HF)in adults.Methods:A prospective study was conducted among the adult patients who were hospitalized in Department of Cardiology,The First Affiliated Hospital of Chongqing Medical University,from September 2022 to May 2024,and the patients who were diagnosed with HF were enrolled as HF group,while those who were excluded from the clinical diagnosis of HF were enrolled as control group.The two groups were compared in terms of general clinical data and related parameters measured by EC,in-cluding stroke volume variation(SVV),systolic time ratio(STR),pre-ejection period(PEP),index of contractility(ICON),ICON varia-tion(VIC),left ventricular ejection time(LVET),and left ventricular ejection fraction(LVEF).A multivariate logistic regression model established by the forward method was used to investigate the factors associated with HF.The receiver operating characteristic(ROC)curve was plotted for the hemodynamic parameters measured by EC in the diagnosis of HF,and the area under the ROC curve(AUC)was calculated.Results:A total of 239 patients were enrolled,with 126 in the HF group and 113 in the control group.Compared with the control group,the HF group had significantly higher SVV,STR,PEP,VIC,and age and significantly lower ICON,LVET,and LVEF(all P<0.001).The multivariate logistic regression model estab-lished by the forward method showed that STR(odds ratio[OR]=1.199,95%CI=1.110-1.294,P<0.001),VIC(OR=1.176,95%CI=1.090-1.269,P<0.001),and age(OR=1.068,95%CI=1.010-1.128,P<0.05)were positively correlated with HF,and ICON(OR=0.968,95%CI=0.941-0.996,P<0.05)and LVEF(OR=0.854,95%CI=0.798-0.913,P<0.001)were negatively correlated with HF.The ROC curve analysis showed that STR,VIC,and ICON had an AUC of 0.887(95%CI=0.848-0.927),0.891(95%CI=0.851-0.932),and 0.718(95%CI=0.654-0.782),respectively,in the diagnosis of HF,with an optimal cut-off value of 37.5%,19.5%,and 49.3,respec-tively,a sensitivity of 91.3%,73.8%,and 50.0%,respectively,and a specificity of 61.4%,93.8%,and 86.7%,respectively.Among these indicators,STR combined with VIC had an AUC of 0.940(95%CI=0.912-0.967)in the diagnosis of HF,with a sensitivity of 83.3%and a specificity of 91.2%.Conclusion:The EC method can effectively assess the cardiac functional status of adult patients,and STR combined with VIC has some clinical value for the diagnosis of heart failure.

OBJECTIVE: To explore the changes of stress and displacement of intervertebral discs and vertebral bodies in adolescent idiopathic cervical kyphosis models caused by Wuqinxi () Huju() and extension movement after torque loading by finite element analysis. METHODS: One healthy male volunteer aged 24-year-old (heighted 178 cm and weighted 65 kg) was selected, software such as Mimics 21.0, Geomagic wrap 2017, SolidWorks 2017, and Ansys Workbench 17.0 were used to simulate adolescent idiopathic cervical spine model, an axial compressive load of 266 N was applied to the center of the end plate on C₂ for head physical gravity simulation, the lower part of C₇ vertebral body was set as the point of freedom constraint, a torque of 1.5 N·m was applied with C₂ as the reference point to simulate the stress on intervertebral discs and vertebral bodies after 45° movement of Wuqinxi () Huju (). RESULTS: The normal C₂-C₇ cervical spine model and adolescent idiopathic cervical kyphosis model were successfully constructed. The maximum stress value of intervertebral disc when the Huju()was raised and extended at 45° and loaded with torque occurred in C_3,4 intervertebral disc (3.588 1) MPa. The maximum stress values of each intervertebral disc were C_3,4(3.588 1 MPa)>C_2,3 (3.467 5 MPa) >C_4,5(2.597 7 MPa) >C_5,6 (2.378 8 MPa) >C_6,7 (1.404 9 MPa), respectively. The maximum stress of C6 vertebral body was 5.842 9 MPa, while the stresses of C₂, C₃, C₄, and C₅ vertebral bodies was 4.184 8, 4.437 8, 4.148 7, and 2.852 4 MPa respectively. The overall stress of vertebral body was mainly concentrated in the front of vertebral body. CONCLUSION After long-term practice of Huju()movement, the stress concentration in intervertebral discs and the front of vertebral body changes the stress load state of intervertebral discs and vertebral body. As time goes by, intervertebral discs may change, forming a shape that is higher in the front and lower in the back. The vertebral body may also undergo remodeling, resulting in a relative increase in the height of the anterior edge of vertebral body and promoting the recovery of cervical kyphosis to a physiological lordosis state.
Humans ; Finite Element Analysis ; Male ; Cervical Vertebrae/physiopathology* ; Kyphosis/therapy* ; Young Adult ; Adolescent ; Adult

OBJECTIVE: To explore the effect of bear bile powder (BBP) on acute lung injury (ALI) and the underlying mechanism. METHODS: The chemical constituents of BBP were analyzed by ultra-high-pressure liquid chromatography-mass spectrometry (UPLC-MS). After 7 days of adaptive feeding, 50 mice were randomly divided into 5 groups by a random number table (n=10): normal control (NC), lipopolysaccharide (LPS), dexamethasone (Dex), low-, and high-dose BBP groups. The dosing cycle was 9 days. On the 12th and 14th days, 20 µL of Staphylococcus aureus solution (bacterial concentration of 1 × 10^-7 CFU/mL) was given by nasal drip after 1 h of intragastric administration, and the mice in the NC group was given the same dose of phosphated buffered saline (PBS) solution. On the 16th day, after 1 h intragastric administration, 100 µL of LPS solution (1 mg/mL) was given by tracheal intubation, and the same dose of PBS solution was given to the NC group. Lung tissue was obtained to measure the myeloperoxidase (MPO) activity, the lung wet/dry weight ratio and expressions of CD14 and other related proteins. The lower lobe of the right lung was obtained for pathological examination. The concentrations of inflammatory cytokines including interleukin (IL)-6, tumour necrosis factor α (TNF-α ) and IL-1β in the bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay, and the number of neutrophils was counted. The colonic contents of the mice were analyzed by 16 sRNA technique and the contents of short-chain fatty acids (SCFAs) were measured by gas chromatograph-mass spectrometer (GC-MS). RESULTS: UPLC-MS revealed that the chemical components of BBP samples were mainly tauroursodeoxycholic acid and taurochenodeoxycholic acid sodium salt. BBP reduced the activity of MPO, concentrations of inflammatory cytokines, and inhibited the expression of CD14 protein, thus suppressing the activation of NF-κB pathway (P<0.05). The lung histopathological results indicated that BBP significantly reduced the degree of neutrophil infiltration, cell shedding, necrosis, and alveolar cavity depression. Moreover, BBP effectively regulated the composition of the intestinal microflora and increased the production of SCFAs, which contributed to its treatment effect (P<0.05). CONCLUSIONS BBP alleviates lung injury in ALI mouse through inhibiting activation of NF-κB pathway and decreasing expression of CD14 protein. BBP may promote recovery of ALI by improving the structure of intestinal flora and enhancing metabolic function of intestinal flora.
Animals ; Acute Lung Injury/pathology* ; Lipopolysaccharides ; Ursidae ; Gastrointestinal Microbiome/drug effects* ; Bile/chemistry* ; Lipopolysaccharide Receptors/metabolism* ; Powders ; Male ; Lung/drug effects* ; Mice ; Peroxidase/metabolism* ; Signal Transduction/drug effects* ; Cytokines/metabolism*

Acute kidney injury (AKI) has high morbidity and mortality, but effective clinical drugs and management are lacking. Previous studies have suggested that macrophages play a crucial role in the inflammatory response to AKI and may serve as potential therapeutic targets. Emerging evidence has highlighted the importance of forkhead box protein O1 (FoxO1) in mediating macrophage activation and polarization in various diseases, but the specific mechanisms by which FoxO1 regulates macrophages during AKI remain unclear. The present study aimed to investigate the role of FoxO1 in macrophages in the pathogenesis of AKI. We observed a significant upregulation of FoxO1 in kidney macrophages following ischemia-reperfusion (I/R) injury. Additionally, our findings demonstrated that the administration of FoxO1 inhibitor AS1842856-encapsulated liposome (AS-Lipo), mainly acting on macrophages, effectively mitigated renal injury induced by I/R injury in mice. By generating myeloid-specific FoxO1-knockout mice, we further observed that the deficiency of FoxO1 in myeloid cells protected against I/R injury-induced AKI. Furthermore, our study provided evidence of FoxO1's pivotal role in macrophage chemotaxis, inflammation, and migration. Moreover, the impact of FoxO1 on the regulation of macrophage migration was mediated through RhoA guanine nucleotide exchange factor 1 (ARHGEF1), indicating that ARHGEF1 may serve as a potential intermediary between FoxO1 and the activity of the RhoA pathway. Consequently, our findings propose that FoxO1 plays a crucial role as a mediator and biomarker in the context of AKI. Targeting macrophage FoxO1 pharmacologically could potentially offer a promising therapeutic approach for AKI.

Objective To establish mouse models exposed to different doses of neodymium oxide via tracheal instillation,and to investigate the mechanisms underlying brain tissue damage induced by neodymium oxide exposure in mice.Methods Forty-eight male C57/BL6 mice were randomly assigned to four groups:the control group,the low-dose group,the medium-dose group,and the high-dose group.The low-dose,medium-dose,and high-dose groups received 62.5 mg/mL,125 mg/mL,and 250 mg/mL neodymium oxide,respectively,via non-exposed tracheal instillation.The control group received an equivalent volume of saline using the same administration method.After 35 days,the mice were euthanized,and brain tissues were collected.RT-PCR was used to assess the mRNA expression changes of Claudin-5 and Occludin.Western blot analysis was performed to evaluate the expression changes of Claudin-5 and Occludin tight junction proteins,as well as the expression changes of MMP-2 and MMP-9 in the brain tissues.Additionally,the expression of the RhoA/ROCK2 signaling pathway and downstream cofilin protein was examined.Changes in oxidative stress markers,including MDA,T-AOC,and NO,were measured using a kit method.Results The mRNA expression of Claudin-5 was significantly reduced in the middle-dose and high-dose groups compared to the control group(P<0.05).Similarly,the mRNA expression of Occludin was significantly lower in the low-dose,medium-dose,and high-dose groups compared to the control group(P<0.05).Additionally,the protein expression of Claudin-5,MMP-2,and Occludin was significantly decreased in the low-dose,medium-dose,and high-dose groups compared to the control group(P<0.05).The protein expression of MMP-9 and RhoA was also signifi-cantly lower in the medium-dose and high-dose groups compared to the control group(P<0.05).Furthermore,the protein expression of ROCK2 and p-cofilin in the high-dose group was significantly lower than that in the control group(P<0.05).The content of MDA and T-AOC was significantly lower in the medium-dose and high-dose groups compared to the control group(P<0.05),and the content of NO in the high-dose group was significantly lower than that in the control group(P<0.05).Conclusion Exposure to neodymium oxide results in increased permeability of the blood-brain barrier in mice,leading to oxidative stress,inflammatory responses,and activation of the RhoA/ROCK2 signaling pathway.