Objective: To analyze the association between insufficient sleep and score of depressive symptoms among junior and senior high school students, so as to provide a scientific reference for targeted early intervention measures of adolescents depressive symptoms. Methods: From September to November 2023, a stratified cluster random sampling method was used to select 96 080 junior and senior high school students from 409 schools in 113 districts and counties in Shaanxi Province. A questionnaire survey was conducted using the 2023 Shaanxi Provincial Common Student Diseases and Health Influencing Factors Survey Form, and their height and weight were measured. Propensity score (PS) matched (1∶1) analysis was used to match participants with insufficient sleep to those sufficient sleep students. Through the gradual correction of the confounders, three multilevel linear models were established to analyze the association between insufficient sleep and depressive symptoms score, and subgroup analysis was conducted afterward. Results: A total of 70 135 (73.00%) students had insufficient sleep. After PS matching, 25 894 pairs were matched. Before PS matching, after adjusting for gender, educational stage, region, adolescent characteristics, boarding status, smoking, alcohol consumption, outdoor activities and body mass index grouping, linear regression analysis results showed that compared with students who got adequate sleep, students who lacked sleep had an increase of 1.39 scores ( B=1.39, 95%CI =1.28-1.51) in depressive symptoms; after PS matching, students with insufficient sleep got an increase of 1.32 scores ( B=1.32, 95%CI =1.17- 1.45 ) in depressive symptoms score compared with those who had adequate sleep (both P <0.05). Conclusions The insufficient sleep is associated with the increase of the depressive symptoms score of junior and senior high school students. It is recommended that junior and senior high school students should keep a good sleeping habit, so as to reduce the prevalence of depressive symptoms.

Objective To generate a dense granule protein 3 (GRA3) gene-deficient mutant of the Toxoplasma gondii ME49 strain and to test the virulence of the mutant. Methods Gene-deficient parasites were generated with the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system. Guide RNA (gRNA) was designed using the E-CRISPR software, and mutated on the pSAG1::Cas9-U6::sgUPRT plasmid using the Q5 site-directed mutagenesis kit to generate the pSAG1::Cas9-U6::sgGRA3 plasmid. A GRA3 donor plasmid containing GRA3 gene upstream sequences, pyrimethamine resistant gene dihydrofolate reductase-thymidylate synthase (DHFR-TS) and GRA3 gene downstream sequence was generated, and GRA3 donor DNA was amplified using PCR assay. The pSAG1::Cas9-U6::sgGRA3 plasmid and GRA3 donor DNA were electroporated into tachyzoites of the wild-type T. gondii ME49 strain. Then, parasite suspensions were inoculated into human foreskin fibroblast (HFF) cells and screened with pyrimethamine to yield pyrimethamine-resistant parasites for monoclonal screening. The GRA3 gene deficient monoclonal strain (ME49Δgra3) of T. gondii was identified using PCR and Western blotting assays, and the expression of GRA3 protein was determined in the T. gondii ME49Δgra3 strain using Western blotting. Subsequently, 1 000 freshly lysed tachyzoites of T. gondii ME49 and ME49Δgra3 strains were transferred to 12-well plates seeded with HFF cells, and incubated at 37 °C containing 5% CO₂ for 7 days, and the number of plaques was counted by staining with crystal violet solutions. HFF cells infected with tachyzoites of T. gondii ME49 and ME49Δgra3 strains were stained using Giemsa solutions, and the numbers of cells containing 1, 2, 4, and > 4 T. gondii parasitophorous vacuoles were counted. In addition, the survival rates of C57BL/6 mice infected with T. gondii ME49 and ME49Δgra3 strains were compared 35 days post-infection. Results PCR assay revealed successful amplification of both the upstream and downstream homologous arm bands of the DHFR-TS gene in the T. gondii ME49Δgra3 strain, and no corresponding bands were amplified in the ME49 strain. The GRA3 band was amplified in the ME49 strain, and the DHFR-TS band, rather than GRA3 band, was amplified in the ME49Δgra3 strain. Western blotting determined absence of GRA3 protein expression in the ME49Δgra3 strain. Crystal violet staining showed that the T. gondii ME49 strain produced more plaques than the ME49Δgra3 strain [(352.67 ± 26.39) plaques vs. (235.00 ± 26.29) plaques; t = 5.472, P < 0.01], and Giemsa staining revealed that the proportion of T. gondii parasitophorous vacuoles containing at least four T. gondii tachyzoites was higher in HFF cells infected with the ME49 strain than in those infected with the T. gondii ME49Δgra3 strain [(75.67 ± 2.52)% vs. (59.67 ± 2.31)%; t = 8.113, P < 0.01], and the proportion of T. gondii parasitophorous vacuoles containing at least 1 or 2 T. gondii tachyzoites was higher in HFF cells infected with the T. gondii ME49 strain than in those infected with the T. gondii ME49Δgra3 strain [(24.33 ± 2.52)% vs. (40.33 ± 2.31)%; t = −8.113, P < 0.01]. In addition, mice infected with the T. gondii ME49 and ME49Δgra3 strains started to die 8 and 9 days post-infection, and the 35-day mortality rates of mice infected with T. gondii ME49 and ME49Δgra3 strains were 10.00% and 70.00% post-infection (χ² = 6.762, P < 0.01). Conclusions The T. gondii ME49Δgra3 strain has been successfully generated, and GRA3 protein may increase the virulence of the T. gondii ME49 strain.

China is a high-incidence region for gastric cancer globally. The disease is characterized by a high morbidity rate, low early diagnostic rate, and poor long-term outcomes, imposing a significant burden on both patients and society. Therefore, exploring the pathogenesis of gastric cancer, developing novel therapeutic strategies, and identifying new drug targets is of great importance. Telomerase expression is broadly associated with cancer cell targeting, and its up-regulation is one of the key factors driving the initiation and progression of gastric cancer. Additionally, telomerase is intricately involved in the regulation of autophagy and autophagy-associated cell death. While autophagy can induce chemoresistance, excessive autophagy may lead to cell death, which also constitutes one of the mechanisms of chemotherapy. Telomerase not only directly contributes to gastric cancer pathogenesis but also indirectly influences its development and treatment by modulating autophagy and autophagic cell death. Therefore, telomerase holds promise as a novel therapeutic target in gastric cancer.
Humans ; Stomach Neoplasms/genetics* ; Telomerase/genetics* ; Autophagy/physiology*

Booster vaccinations are highly recommended in combating the SARS-CoV-2 Omicron variant and its subvariants. However, the optimal booster vaccination strategies and related immune mechanisms with different prior vaccinations are under-revealed. In this study, we systematically evaluated the immune responses in mice and hamsters with different prime-boost regimens before their protective efficacies against Omicron were detected. We found that boosting with Ad5-nCoV, S_WT-2P or S_Omicron-6P induced significantly higher levels of neutralization activities against Omicron variants than CoronaVac and ZF2001 by eliciting stronger germinal center (GC) responses. Specifically, S_Omicron-6P induced even stronger antibody responses against Omicron variants in CoronaVac and Ad5-nCoV-primed animals than non-Omicron-specific vaccines but with limited differences as compared to Ad5-nCoV and S_WT-2P. In addition, boosting with a specific vaccine has the potential to remodel the existing immune profiles. These findings indicated that adenovirus-vectored vaccines and mRNA vaccines would be more effective than other types of vaccines as booster shots in combating Omicron infections. Moreover, the protective efficacies of the vaccines in booster vaccinations are highly related to GC reactions in secondary lymphatic organs. In summary, these findings provide timely important information on prime-boost regimens and future vaccine design.

Objective To investigate the feasibility of simultaneous multi-slice(SMS)acquisition combined with single-shot echo-planar imaging(SSEPI)multi-model diffusion weighted imaging(DWI)for breast lesions.Methods Totally 108 cases of breast lesions were retrospectively enrolled and divided into malignant group(n=66)and benign group(n=42)based on pathology.3.0T MR scanner was used to acquire SSEPI and SMS-SSEPI multi-b values DWI,7 derived parameters were obtained through post-processing with mono-exponential,fractional-order calculus(FROC)and continuous-time random walk(CTRW)models.Then the imaging quality and derived parameters of SMS-SSEPI and SSEPI DWI were compared between groups.Spearman correlation analysis was performed to explore the relationships of corresponding parameters between SMS-SSEPI DWI and SSEPI DWI.Diagnostic performance of each parameter for distinguishing malignant and benign lesions was evaluated according to the area under the receiver operating characteristic curve(AUC).Results Background noise score of SMS-SSEPI DWI was lower than that of SSEPI DWI(P＜0.05),whereas no significant difference of overall imaging quality,normal anatomical structure depiction,lesion conspicuity,geometric distortion,signal-to-noise ratio(SNR)nor contrast-to-noise ratio(CNR)was found between SMS-SSEPI DWI and SSEPI DWI(all P＞0.05).Parameters derived from SMS-SSEPI DWI were all moderately to highly positively correlated with those from SSEPI DWI(rs=0.66-0.98).Malignant lesions exhibited significantly lower apparent diffusion coefficient(ADC),diffusion coefficient based on FROC(DFROC),fractional order derivative in space(βFROC),diffusion coefficient based on CTRW(DCTRW),temporal diffusion heterogeneity index(αCTRW)and spatial diffusion heterogeneity index(βCTRW)values,but higher spatial parameter(μFROC)value than benign lesions(all P＜0.05).AUC of SMS-SSEPI DWI derived parameters for differentiating malignant from benign lesions were 0.699-0.900,of those from SSEPI DWI were 0.654-0.887,while in both SMS-SSEPI DWI and SSEPI DWI,DFROC had the highest diagnostic efficacy(AUC=0.900,0.887).Conclusion SMS-SSEPI DWI could be used to effectively differentiate malignant and benign breast lesions.

Objective To investigate the effects of safflower polysaccharide on tumor growth and the Phos-phoinositide 3-kinase(PI3K)/Protein Kinase B(Akt)/Mammalian Target of Rapamycin(mTOR)signaling pathway in colorectal cancer mice.Methods The mouse model of colorectal cancer was established by inoculating SW480 colorectal cancer cells.The successfully modeled mice were divided into five groups:the control group,the low-dose safflower polysaccharide group 15 mg/(kg·day),the medium-dose safflower polysaccharide group 45 mg/(kg·day),and the high-dose safflower polysaccharide group 135 mg/(kg·day).After treatment,tumor volume,mass,and tumor inhibition rate were measured.Immunohistochemistry was used to detect the expression of Proliferating Cell Nuclear Antigen(PCNA)protein.Apoptosis was assessed using the Terminal deoxynucleotidyl Transferase-Mediated Nick End Labeling(TUNEL)method.Hematoxylin-Eosin(HE)staining was performed to observe the morphology of tumor tissues.Real-time polymerase chain reaction(RT-PCR)was employed to measure the expression levels of PI3K,Akt,and mTOR in tumor tissues from each group.Western blot analysis was conducted to evaluate the expres-sion levels of c-Myc,Bcl-2-associated X protein(Bax),and proteins related to the PI3K/Akt/mTOR pathway.Results Compared with the model group,the safflower polysaccharide(low,medium,and high)dose groups exhibited significantly decreased tumor mass and volume,as well as reduced mRNA levels of PI3K,Akt,and mTOR.Additionally,the expression ratios of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,and c-Myc protein were notably decreased,while the expression of Bax protein was significantly increased(P<0.05).As the concentration of safflower polysaccharide increased,the tumor inhibition rate also increased(P<0.05).In the model group,tumor tissue from mice showed a higher number of PCNA-positive cells;TUNEL staining revealed minimal green fluores-cence,indicating low apoptosis rates.The tumor cells were closely packed,large in size,and poorly differentiated.Conversely,in the different dosage safflower polysaccharide groups,the expression of PCNA in tumor tissues was markedly reduced,and TUNEL staining positive cells(green)were significantly increased,leading to higher apopto-sis rates(P<0.05).The tumor tissue cells were loosely arranged,with shrunken nuclei and numerous necrotic cells.Conclusion Safflower polysaccharide potentially inhibits the growth of colorectal cancer in mice through suppression of the PI3K/Akt/mTOR signaling pathway.

The left maxilla of a patient was resected because of tumor,and the defect was reconstruted with fibular transplantation.The autonomous dental implant robot technology was used to achieve precise implantation of multiple implants and immediate denture restoration within the limited left maxilla.The surgery was minimally invasive and efficient,and significantly reducing patient post-operative discomfort.The final restoration was completed 6 months after surgery.

Objective To construct a global model of cardiac surface motion based on average intensity projection(AIP)image.Methods The cine magnetic resonance imaging data were divided into training set and test set for model construction and validation.The datum points were obtained on the AIP surface point cloud,and the corresponding points of each cardiac phase surface point cloud were found based on surface features.Principal component analysis was used to extract feature information,establish the mapping relationship between the datum points and the corresponding points,and construct a model for predicting each temporal phase surface point cloud from the AIP surface point cloud.Results The RMSE of the model on test set for corresponding point prediction ranged from(0.209±0.020)mm to(0.841±0.074)mm,while the Euclidean distance for each time phase surface point cloud prediction ranged from(1.399±0.029)mm to(1.658±0.100)mm.Conclusion The proposed global model exhibit high accuracy and can provide a reference for image segmentation and clinical treatments.

Objective To develop a combined model integrating radiomics and 3D deep learning features for improving the predictive efficacy of overall survival in non-small cell lung cancer(NSCLC)patients undergoing radiotherapy,thereby providing a foundation for optimizing individualized radiotherapy strategies.Methods A retrospective analysis was conducted on 522 NSCLC patients from 3 centers.Radiomics features were extracted from the tumor region of interest on radiotherapy planning CT scans,and a 3D-SE-ResNet was constructed to extract deep learning features.Following feature extraction,features were selected via univariate Cox analysis and Lasso-Cox regression,and a combined model was established by fusing the two feature types through principal component analysis.The discriminative ability of the model was evaluated using the concordance index(C-index)and the area under the receiver operating characteristic curve(AUC),while the risk stratification efficacy was verified by Kaplan-Meier survival analysis.Results The predictive performance of deep learning features was significantly superior to that of radiomics features(C-index:0.73 vs 0.65).The combined model achieved the highest predictive performance in the training set,internal test set,and external test set(C-index:0.74,0.69,0.72 respectively),with higher AUC values for predicting 1-year,2-year,and 3-year OS than either single model.Kaplan-Meier analysis showed significant differences in survival between the high-and low-risk groups(Log-rank test,P<0.001),and calibration curves indicated good consistency between predicted and actual survival outcomes.Conclusion The combined model integrating radiomics and 3D deep learning features can accurately predict survival outcomes in NSCLC patients undergoing radiotherapy.The multi-center validation results support its potential application in prognosis stratification for individualized radiotherapy.

Objective To evaluate the diagnostic efficacy of sound touch viscoelastography(STVi)and shear wave elastography(SWE)in distinguishing between benign and malignant breast nodules.Methods A total of 104 breast nodules(52 benign and 52 malignant)from 102 patients scheduled for surgical treatment at Binzhou Medical University Hospital between October 2024 and February 2025 were prospectively enrolled.All nodules were pathologically confirmed through surgical excision or core needle biopsy.The viscosity coefficient and Young's modulus of both intranodular and perinodular tissues within a 2-mm range were measured using the Mindray Resona A20S ultrasound system.The diagnostic performance of each parameter,the correlation between elastic parameter values and the maximum nodule diameter,as well as the inter-correlation between the two parameters were systematically analyzed.Results The elasticity parameters were significantly higher in malignant nodules[maximum intranodular Viscosity coefficient(Vimax):5.93(4.33,8.47)Pa·s,maximum Young's modulus(Emax):81.18(58.31,120.33)kPa;maximum Viscosity coefficient of the surrounding 2-mm tissue(Vi2max):7.57(5.40,10.16)Pa·s,maximum Young's modulus(E2max):117.21(65.66,170.66)kPa]compared to benign nodules[Vimax:3.70(2.69,5.32)Pa·s,Emax:41.42(28.29,64.25)kPa;Vi2max:4.30(3.63,5.65)Pa·s,E2max:47.23(36.42,74.67)kPa](P<0.05).The diagnostic performance of the 2-mm perinodular tissue(Vi2max:0.78,E2max:0.81)surpassed that of intranodular tissue(Vimax:0.72,Emax:0.77)(P<0.05).The combined diagnostic model(Vi2+E2,Vi+E)achieved AUC values of 0.82 and 0.77,respectively,which outperformed STVi alone(P<0.05)and showed marginally better performance than SWE alone,although the difference was not statistically significant(P>0.05).The maximum nodule diameter showed a moderate correlation with the elasticity parameters,with E2max exhibiting the strongest correlation(r=0.510,P<0.05).Conclusions Both STVi and SWE show clinical value in distinguishing between benign and malignant breast nodules.Particularly,elasticity parameters obtained from the 2-mm perinodular tissue demonstrate better diagnostic performance than those measured within the nodule itself,and combining these parameters enhances the overall diagnostic accuracy of STVi.