ObjectiveTo explore the relationship between negative life events and depression severity in adolescent patients with depressive disorder， as well as the chain mediating role of insomnia symptoms and quality of life. Methods374 outpatient patients and hospitalized patients with adolescent depressive disorders were enrolled. The Adolescent Life Event Scale （ASLEC）， the Insomnia Severity Index （ISI）， the World Health Organization Quality of Life Questionnaire Short Form （WHOQOL-BREF）， and the Center for Epidemiology Depression Scale （CES-D） were used to evaluate the negative life event situation， insomnia symptoms， quality of life level and depression severity of the subjects， respectively. In addition， the PROCESS 4.0 macroprogram was used to analyze the chain mediating effect of insomnia symptoms and quality of life between negative life events and depression severity in patients with adolescent depressive disorder. ResultsThe results of correlation analysis showed that there was a significant correlation between negative life events and insomnia symptoms， quality of life， and depression severity （all P<0.05）. In addition， the results of chain mediation showed that negative life events had a significant direct effect on depression severity， with an effect size of 0.12 （P<0.001）. Insomnia symptoms and quality of life played a mediating role in the relationship between negative life events and depression severity in patients with adolescent depressive disorders， with indirect effect sizes of 0.062 （95%CI： 0.040-0.087） and 0.091 （95%CI： 0.059-0.123）， respectively. It could also play a chain mediation role， and the effect size was 0.039 （95%CI： 0.024-0.057）. ConclusionNegative life events experienced by patients with adolescent depressive disorder not only directly affect the severity of depressive symptoms， but may also indirectly exacerbate depression through insomnia symptoms and quality of life.

Zoledronic acid (ZA), a nitrogen-containing bisphosphonate with established clinical utility in osteoporosis management, exhibits emerging antitumor potential in estrogen receptor-positive breast cancer. However, the molecular mechanisms underlying its nonapoptotic anticancer effects remain poorly characterized. This study revealed that ZA induced ferroptosis in ER+ breast cancer cells through dual suppression of cystine-glutamate antiporter SLC7A11 and glutathione peroxidase 4 (GPX4), key repressors of ferroptosis. Pharmacological inhibition of ferroptosis using Ferrostatin-1 significantly attenuated ZA-induced cytotoxicity, while combinatorial treatment with the GPX4 inhibitor RSL3 synergistically enhanced lipid peroxidation and cell death. Mechanistically, ZA activated the Hippo-YAP signaling pathway, promoting YAP phosphorylation, proteasomal degradation, and cytoplasmic retention, thereby silencing SLC7A11 and GPX4. We established a novel metabolic vulnerability in hormone-responsive malignancies.These findings position ZA as a bifunctional ferroptosis inducer in ER+ breast cancer, offering a promising strategy to overcome endocrine resistance.

Objective:To investigate the characteristics of fatty acid metabolism in the hepatic steatosis (HS) population and to explore the role of inflammation in the relationship between fatty acid metabolism and HS.Methods:A total of 3 112 subjects from the 2011–2014 National Health and Nutrition Examination Survey (NHANES) database were included in this retrospective study, among whom 1 552 were in the HS group (weighted percentage: 49.87%) and 1 560 in the control group (weighted percentage: 50.13%). The differences in dietary fatty acid and serum free fatty acid (FFA) levels and metabolic characteristics were compared. Logistic and weighted quantile sum (WQS) regression models were used to evaluate the impacts of various FFA indexes on HS. Mediation analysis was conducted to examine whether inflammation served as a mediator in the relationship between FFA and HS.Results:Dietary fatty acid intake did not differ significantly between the HS and control groups. However, serum FFA metabolism characteristics differed between these two groups, with the concentrations of 28 FFAs in the HS group significantly higher than those in the control group. The risk of HS was correlated negatively with serum unsaturated fatty acids/saturated fatty acids ( OR=0.69, 95% CI: 0.64–0.75, P<0.001), polyunsaturated fatty acids/saturated fatty acids ( OR=0.63, 95% CI: 0.99–0.69, P<0.001), and Health Promotion Index ( OR=0.78, 95% CI: 0.74–0.82, P<0.001) but positively with thrombosis index ( OR=2.20, 95% CI: 1.89–2.55, P<0.001). The WQS model showed that behenic acid had the greatest contribution weight to increase the risk of HS among the 30 FFAs. After adjusting for confounding factors, the risk of HS increased by 167% (95% CI: 2.14–3.34, P<0.001) for each unit increase in the WQS index. Further mediation analysis showed that albumin/globulin ratio, white blood cell count, and neutrophil to platelet ratio significantly mediated the relationship between serum FFA and HS ( P<0.05). Conclusion:People with similar dietary fatty acid structures may present different serum FFA metabolism characteristics. Serum FFA level correlates with the risk of HS, and inflammation plays an important role in mediating the relationship between fatty acid metabolism and HS.

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.

In vaccination,traditional vaccines are administered intramuscularly or subcutaneously,mainly inducing humoral immunity,with weak induction of mucosal immunity and insufficient protection against respiratory and gastrointestinal viruses.Current research has found that immuni-zation vaccines can effectively induce mucosal immunity through oral or nasal administration,but there is a lack of effective mucosal adjuvants,and there is an urgent need for the development of safe and effective new mucosal adjuvants.With the advancement of nano-adjuvant research,novel nanoparticles have attracted widespread attention due to their good mucosal adhesion,efficient an-tigen-loading ability,biocompatibility,and advantages in immune-enhancement,nano-size effect,tissue-targeting,and slow release of antigens,etc.,and are expected to be used as novel mucosal ad-juvants in the prevention and treatment of animal diseases.To date,significant progress has been made in research targeting mucosal adjuvants such as polymeric nanoparticles,inorganic nanoparti-cles,and liposomes.This paper reviews the application of nanomucosal adjuvants in animal vaccines and their mechanisms of action at home and abroad,and analyzes the physicochemical factors af-fecting the immune-enhancing effect of nanoparticle adjuvants,with a view to providing a reference for the development of novel mucosal adjuvants.

Objective:To explore the construction and efficacy of cranial magnetic resonance imaging(MRI)classification model based on attention mechanism in type 2 diabetes patients with cognitive impairment.Methods:The case data of 100 patients with type 2 diabetes who were treated in the General Hospital of North China Petroleum Administration Bureau from June 2022 to January 2024 were retrospectively selected.A total of 100 MRI images of cranial FLAIR_LongTR sequence with cognitive impairment(32 cases)and those without cognitive impairment(68 cases)were respectively collected.The images of the above two kinds of samples were horizontally and vertically translated to expand to 1000 samples,respectively.The samples were randomly divided into training samples(n=700)and test samples(n=300)as the ratio of 7:3 according to affine transformation data augmentation method.Then,the attention mechanism model was established to test the images with full scan of the test samples.The ability of the attention mechanism system in screening cognitive impairment was analyzed according to the method of setting threshold value.The 100 MRI images of cranial FLAIR_LongTR sequence of patients in our hospital from January 2024.From January to May 2024 were used as a verification set to verify the diagnostic value of attention mechanism.Results:With the increasing of iteration times,the sample loss of training and verification of attention mechanism model gradually decreased and tended toward stability,and the accuracy of training set and verification set gradually increased and tended toward stability.In the attention mechanism model,the average loss rate of training samples was 10.024%,and that of test samples was 15.247%.In the attention mechanism model,the average accuracy of training samples was 99.078%,and the average accuracy of test samples was 99.753%.Receiver operating characteristic(ROC)curves showed that the area under curve(AUC)of attention mechanism model was 0.998,which can better diagnose cognitive impairment of patients with type 2 diabetes than the resNET model(AUC=0.656)(Z=3.437,P<0.001).Conclusion:The constructed cranial MRI classification model by using attention mechanism has favorable diagnostic value for cognitive impairment in patients with type 2 diabetes.

The aim of this study was to qualitatively and quantitatively detect coronavirus(CoV)in the feces of bats from Qinghua Cave,Yunnan Province,China.CoV was qualitatively tested with reverse transcription polymerase chain reaction(RT-PCR),and homology and genetic evolution were analyzed with bioinformatics software.The established reverse transcription real-time fluores-cence quantitative PCR(qRT-PCR)method was applied to CoV quantification in bat feces.The positivity rate of CoV in 306 fecal samples collected from the fulvous fruit bat(Rousettus leschenaultia)was 7.8%(24/306)according to RT-PCR.All 24 strains of CoV belonged to β-CoV,and showed a similarity of 86.8%-100.0%at the nucleotide level and 95.2%-100.0%at the amino acid level,with respect to other β-CoV sequences in the NCBI database.The positivity rate of CoV was 18.6%(57/306)according to qRT-PCR,a value higher than that according to RT-PCR(χ2=25.3,P<0.05).The mean β-CoV load was 1.3×103 copies/μL.In conclusion,the bats in Qinghua Cave,Yunnan Province,carried CoV belonging to β-CoV.The established qRT-PCR method achieved good sensitiv-ity,accuracy,reproducibility,and a higher detection rate than that of RT-PCR,and can be used for rapid detection of β-CoV in bats.

BACKGROUND:Recent studies have shown that Du Meridian electroacupuncture has a unique effect on alleviating spinal cord injury,but the underlying mechanisms require further clarification.OBJECTIVE:To investigate the regulatory effects and the associated action mechanisms of Du Meridian electroacupuncture on ferroptosis after cervical spinal cord injury in rats.METHODS:One hundred SD rats were randomly divided into sham,model,Du Meridian electroacupuncture,RSL3,and Du Meridian electroacupuncture+RSL3 groups.The sham group underwent only laminectomy.The other four groups were subjected to cervical spinal cord injury by the Allen method.The Du Meridian electroacupuncture group received electroacupuncture after cervical spinal cord injury.The RSL3 group received intraperitoneal injections of glutathione peroxidase 4 inhibitor RSL3 after cervical spinal cord injury.The Du Meridian electroacupuncture+RSL3 group received both electroacupuncture and RSL3 intervention after cervical spinal cord injury.Samples were collected on postoperative days 7 and 28 to assess motor function,histological morphology,neuronal survival,glial scar formation,oxidative stress levels,Fe2+content,glutathione peroxidase 4,and long-chain acyl-CoA synthetase 4 expression.RESULTS AND CONCLUSION:(1)Finally,90 rats completed the follow-up experiment,with 18 rats in each group.(2)FLS and BBB scores were significantly higher in the Du Meridian electroacupuncture group compared with the model and Du Meridian electroacupuncture+RSL3 groups(P＜0.05).(3)Compared with the model group,Du Meridian electroacupuncture improved cervical spinal cord tissue morphology and mitochondrial ultrastructure,while these effects were inhibited by RSL3.(4)Du Meridian electroacupuncture increased the expression of microtubule-associated protein 2,glutathione peroxidase 4,glutathione,and superoxide dismutase(P＜0.05)and reduced the expression of glial fibrillary acidic protein,long-chain acyl-CoA synthetase 4,reactive oxygen species,malondialdehyde,and Fe2+compared with the model group(P＜0.05).However,RSL3 reversed the inhibitory effects of Du Meridian electroacupuncture on ferroptosis,lipid peroxidation and oxidative stress.(5)The results suggest that Du Meridian electroacupuncture inhibits ferroptosis by regulating the glutathione peroxidase 4/long-chain acyl-CoA synthetase 4 axis,thereby reducing secondary neuronal damage and glial scar formation after cervical spinal cord injury and improving neurological function.

AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.

Objective:To investigate the characteristics of fatty acid metabolism in the hepatic steatosis (HS) population and to explore the role of inflammation in the relationship between fatty acid metabolism and HS.Methods:A total of 3 112 subjects from the 2011–2014 National Health and Nutrition Examination Survey (NHANES) database were included in this retrospective study, among whom 1 552 were in the HS group (weighted percentage: 49.87%) and 1 560 in the control group (weighted percentage: 50.13%). The differences in dietary fatty acid and serum free fatty acid (FFA) levels and metabolic characteristics were compared. Logistic and weighted quantile sum (WQS) regression models were used to evaluate the impacts of various FFA indexes on HS. Mediation analysis was conducted to examine whether inflammation served as a mediator in the relationship between FFA and HS.Results:Dietary fatty acid intake did not differ significantly between the HS and control groups. However, serum FFA metabolism characteristics differed between these two groups, with the concentrations of 28 FFAs in the HS group significantly higher than those in the control group. The risk of HS was correlated negatively with serum unsaturated fatty acids/saturated fatty acids ( OR=0.69, 95% CI: 0.64–0.75, P<0.001), polyunsaturated fatty acids/saturated fatty acids ( OR=0.63, 95% CI: 0.99–0.69, P<0.001), and Health Promotion Index ( OR=0.78, 95% CI: 0.74–0.82, P<0.001) but positively with thrombosis index ( OR=2.20, 95% CI: 1.89–2.55, P<0.001). The WQS model showed that behenic acid had the greatest contribution weight to increase the risk of HS among the 30 FFAs. After adjusting for confounding factors, the risk of HS increased by 167% (95% CI: 2.14–3.34, P<0.001) for each unit increase in the WQS index. Further mediation analysis showed that albumin/globulin ratio, white blood cell count, and neutrophil to platelet ratio significantly mediated the relationship between serum FFA and HS ( P<0.05). Conclusion:People with similar dietary fatty acid structures may present different serum FFA metabolism characteristics. Serum FFA level correlates with the risk of HS, and inflammation plays an important role in mediating the relationship between fatty acid metabolism and HS.