Objective: To investigate follow-up participation situation, negative conversion, and tuberculosis incidence among boarding junior and senior high school freshmen with latent tuberculosis infection(LTBI) in Zhuhai, so as to provide a reference for better LTBI management in school settings. Methods: A prospective cohort study enrolled 413 LTBI and 27 184 non LTBI boarding junior and senior high school freshmen in Zhuhai from September to October 2023. The LTBI group underwent follow up retesting at >3-6, >6-12, >12-24, >24 months after screening. The followup content included mycobacterium tuberculosis infection testing,chest X ray examination,and questionaire survey(basic demographic information, lifestyle, history of close contact with tuberculosis, and suspected symptoms of pulmonary tuberculosis). Follow-up participation, negative conversion, and tuberculosis incidence were analyzed. Rate was compared by using the Chi-square test, and factors associated with negative conversion were assessed by multivariate Logistic regression analysis. Results: Follow-up participation rates increased initially and then declined, with rates of 25.67% within >3-6 months, 37.05% within >6-12 months, 82.02% within >12-24 months, and 52.21% after 24 months. The overall negative conversion rate was 61.03%. The differences in the conversion rate to negative of mycobacterium tuberculosis infection detection among junior and senior high school freshmen in the LTBI group by different testing methods, school types, smoking history, sleep status, and academic stress were all statistically significant( χ 2=124.96, 22.70, 4.71, 6.31, 6.08, all P <0.05). Multivariate Logistic regression analysis identified testing methods, smoking history, and sleep status as factors associated with negative conversion ( OR =1.90-21.67, all P <0.05). The cumulative incidence of tuberculosis was 726.39 per 100 000 in the LTBI group versus 14.71 per 100 000 in the non-LTBI group ( RR =49.37, 95% CI =11.08-219.87, P <0.01). Conclusions Boarding junior and senior high school freshmen with LTBI in Zhuhai have a higher risk of tuberculosis than concurrent non-LTBI students; they therefore represent a priority group for tuberculosis prevention and control. Centralized follow up is recommended to improve participation, and high specificity methods should be used in follow up retesting and maintain consistency across assessments.

ObjectiveTo investigate whether photoreceptor-specific knockout of lanosterol synthase （Lss） induces age-related retinal degeneration and to elucidate the underlying molecular mechanisms. MethodsPhotoreceptor-specific Lss knockout mice （Lss^flox/flox； Rho-Cre， CKO） models were established. Retinal structure and function were assessed at 1， 4， and 8 months of age using fundus photography， optical coherence tomography （OCT）， scotopic electroretinogram （ERG） and hematoxylin and eosin （HE） staining. The mouse photoreceptor cell line 661W served as an in vitro model. Lss was knocked down via siRNA， followed by analyses including Seahorse XF mitochondrial stress test， JC-1， MitoSOX Red and DCFH-DA probes to evaluate mitochondrial function and oxidative stress， glycolytic rate assay for glycolysis ability， RT-qPCR for inflammatory factors， γH2AX immunofluorescence for double-strand DNA damage detection， and transcriptome sequencing to identify differentially expressed genes and enriched pathways. ResultsCompared with controls， 1-month-old CKO mice showed no abnormalities. Mild fundus lesions appeared at 4 months. By 8 months， CKO mice exhibited significant thinning of the outer nuclear layer， disorganization of the inner/outer segment structure， and markedly reduced amplitudes of the dark-adapted ERG a- and b-waves （P<0.05）. Transcriptome analysis revealed 1 842 differentially expressed genes （826 up-regulated， 1 016 down-regulated）， with Lss being the most significantly downregulated gene； these genes significantly enriched in sterol biosynthesis， oxidative phosphorylation， aerobic electron transport chain， and ATP synthesis （P<0.05）. In 661W cells， knockdown of Lss significantly reduced mitochondrial basal respiration （P=0.006）， ATP production （P=0.015）， maximal respiration （P<0.001）， and spare respiratory capacity （P<0.001）， while basal glycolysis （P=0.005） and compensatory glycolysis （P=0.028） were increased. These changes were accompanied by mitochondrial membrane potential depolarization （P<0.001）， elevated mitochondrial reactive oxygen species （ROS） levels （P<0.001）， and increased total ROS levels （P=0.013）. Additionally， the mRNA expression of inflammatory factors， including Ccl2 （P=0.004）， Ccl5 （P=0.012）， Cxcl1 （P<0.001）， Cxcl5 （P<0.000 1）， Tgfb （P=0.008）， Hgf （P<0.000 1） and Mmp3 （P=0.003） was significantly upregulated. Moreover， γH2AX fluorescence intensity was increased （P=0.002）. ConclusionsPhotoreceptor-specific Lss deficiency induces mitochondrial dysfunction， oxidative stress and cellular inflammation， ultimately leading to age-dependent retinal degeneration. This study provides novel mechanistic insights and potential therapeutic targets for hereditary and age-related retinal degenerative diseases.

Objective To observe the therapeutic effects and mechanisms of acupuncture at"Yanglingquan"(GB34)and"Zusanli"(ST36)on knee osteoarthritis(KOA)rats,focusing on the regulation of PTEN-induced kinase 1/Parkin(PINK1/Parkin)-mediated mitophagy in chondrocytes.Methods Ten out of 40 male SD rats were randomly selected as the normal group,and the remaining 30 rats were used to construct a KOA model.A total of 27 rats were successfully modeled.The successfully modeled rats were randomly divided into a model group,an acupuncture group(acupuncture at Yanglingquan and Zusanli),and an acupuncture+autophagy inhibitor(3-MA)group(acupuncture at Yanglingquan and Zusanli combined with intraperitoneal injection of 3-MA),with 9 rats in each group.After intervention,the morphology of cartilage tissue in each group was observed,and the Mankin's score of cartilage was assessed.The apoptosis rate of chondrocytes was calculated.Enzyme-linked immunosorbent assay(ELISA)was used to detect inflammatory factors[cyclooxygenase 2(COX-2),interleukin 1β(IL-1β),and tumor necrosis factor α(TNF-α)],oxidative damage indicators[nitric oxide(NO),superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)],and the levels of matrix metalloproteinase 3(MMP-3)and tissue inhibitor of metalloproteinase 1(TIMP-1).The DHE probe was used to measure reactive oxygen species(ROS)level.Polymerase chain reaction(PCR)was used to detect the mRNA expression levels of PTEN-induced kinase 1(PINK1),Parkin,and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)in cartilage tissue.Western blot was used to measure the protein expression levels of PINK1,Parkin,and LC3-Ⅱ in cartilage tissue.Results Compared with the normal group,the model group showed increased Mankin's score,apoptosis rate,COX-2 level,IL-1β and TNF-α levels,NO and MDA levels,MMP-3 and TIMP-1 levels,and ROS content,as well as decreased SOD and GSH-Px levels,and reduced mRNA and protein expressions of PINK1,Parkin and LC3-Ⅱ,with statistically significant differences(P＜0.05).Compared with the model group,the acupuncture group showed decreased Mankin's score,apoptosis rate,COX-2 level,IL-1β and TNF-α levels,NO and MDA levels,MMP-3 and TIMP-1 levels,and ROS content,as well as increased SOD and GSH-Px levels,and elevated mRNA and protein expressions of PINK1,Parkin and LC3-Ⅱ,with statistically significant differences(P＜0.05).Compared with the acupuncture group,the acupuncture+3-MA group showed increased Mankin's score,apoptosis rate,COX-2 level,IL-1β and TNF-α levels,NO and MDA levels,MMP-3 and TIMP-1 levels,and ROS content,as well as decreased SOD and GSH-Px levels,and mRNA and protein expressions of PINK1,Parkin and LC3-Ⅱ,with statistically significant differences(P＜0.05).Conclusion Acupuncture at"Yanglingquan"and"Zusanli"can inhibit chondrocyte apoptosis by activating PINK1/Parkin-mediated mitophagy,thus improving the condition of KOA rats.

Objective To investigate how culture duration affects the expression of immune membrane proteins in human monocyte-derived dendritic cells (DCs) and their exosomes (DEXs). Methods Human monocytes were induced with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) to differentiate into DCs and were subsequently matured with tumor necrosis factor α(TNF-α). Exosomes were isolated by ultracentrifugation, and DEXs were identified by transmission electron microscopy and Amnis imaging flow cytometry, which were also used to quantify the expression of immune membrane proteins on DCs and DEXs. Results On the 10th day of culture, DCs displayed high surface expression of CD11c, CD80, CD86, major histocompatibility complex class I (MHC-I), and MHC-II. Expression peaked at day 18(CD11c: 78.66%±20.33%, CD80: 76.41%±10.02%, CD86: 96.43%±0.43%, MHC-I: 84.71%±2.96%, MHC-II: 80.01%±7.03%). After day 24, the overall expression showed a declining trend, with statistically significant differences observed for all markers except CD80 and MHC-II. By day 30, 80% of the DCs still expressed CD80, CD86, and MHC-II. The expression of immune membrane proteins on DEX surfaces also reached its peak on day 18, followed by an overall decline with prolonged culture time, with statistically significant differences observed for all markers except CD80. Correlation analysis revealed a significant positive relationship between the expression levels of immune membrane proteins on DC and DEX surfaces (CD11c: r=0.98; CD80: r=0.65; CD86: r=0.82; MHC-I: r=0.86; MHC-II: r=0.93). Conclusion Human monocyte-derived DCs in vitro express high expression of immune membrane proteins and maintain stable expression over a specific period. The exosomes secreted by these cells similarly demonstrate high surface expression of immune membrane proteins, with temporal trends aligned with those of the parent DCs.
Humans ; Dendritic Cells/immunology* ; Exosomes/immunology* ; Monocytes/metabolism* ; Cells, Cultured ; Time Factors ; B7-1 Antigen/metabolism* ; Membrane Proteins/immunology* ; Cell Culture Techniques/methods* ; B7-2 Antigen/metabolism* ; Cell Differentiation ; CD11c Antigen/metabolism* ; Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology*

Objective To investigate the factors influencing the efficacy of intravesical Bacille Calmette-Guérin(BCG)instillation after transurethral resection of bladder tumor(TURBT)in patients with intermediate-and high-risk non-muscle invasive bladder cancer(NMIBC),and to construct a prediction model for recurrence after BCG treatment.Methods A retrospective cohort study was conducted on the subjected patients diagnosed with intermediate-and high-risk NMIBC undergoing TURBT followed by standard BCG instillation.The 110 patients treated in Department of Urology of Army Medical Center of PLA from January 2018 to December 2023 were assigned into a training set,while the 52 patients treated at Department of Urology of General Hospital of Central Theater Command from January 2015 to December 2020 were into an external validation set.A total of 17 variables were included and analyzed.Univariate and multivariate Cox regression analyses were performed to identify factors associated with recurrence after BCG instillation,and nomograms were plotted to predict 1-year,3-year,and 5-year recurrence-free survival(RFS).Calibration curve,decision curve analysis(DCA),and receiver operating characteristic(ROC)curve analysis were conducted for internal and external validation to evaluate the predictive performance and clinical utility of the model.Results In the training set,26 patients(23.64%)experienced recurrence during the follow-up period,with a median RFS of 32.00(18.00～50.50)months.Univariate Cox regression analysis suggested that platelet count,eosinophil to lymphocyte ratio(ELR),neutrophil to lymphocyte ratio(NLR),platelet to lymphocyte ratio(PLR),systemic immune inflammation(SII)index,and neutrophil-monocyte to lymphocyte ratio(NMLR),pathological T1 stage(pT1)tumor and hemoglobin,albumin,lymphocyte,and platelet(HALP)score were potential factors influencing recurrence after BCG instillation.Multivariate Cox regression analysis identified high HALP score(HR=0.185,95%CI:0.046～0.736,P=0.017)as an independent protective factor,while high ELR(HR=3.599,95%CI:1.505～8.608,P=0.004)and pT1 stage(HR=3.240,95%CI:1.191～8.818,P=0.021)were independent risk factors for recurrence.Based on this,a nomogram prediction model was constructed.The calibration curves demonstrated good agreement between predicted and actual 1-,3-,and 5-year recurrence risks.Decision curve analysis indicated clinical utility across a wide threshold probability range.In the training set,the model showed strong predictive performance for 1-(AUC=0.842),3-(AUC=0.847),and 5-year(AUC=0.887)recurrence risks,which was further validated in the external cohort.Conclusion Higher HALP score prior to BCG instillation therapy is a protective factor against tumor recurrence,while higher ELR and pT1 stage are risk factors.Our nomogram prediction model based on HALP score,ELR and pathological T stage,can identify individuals at high risk of recurrence after BCG instillation therapy.

Objective To investigate risk factors for residual lesions after initial transurethral resection of bladder tumors(TURBT)and risk factors for tumor recurrence after second TURBT in patients with non-muscle-invasive bladder cancer(NMIBC)in order to provide reference for clinical management.Methods A case-control study design was adopted to include 120 NMIBC patients who underwent initial TURBT and then second surgery within 2～8 weeks in our department from January 2017 to January 2025.Based on the presence of residual lesions after the initial TURBT or not,the patients were divided into a residual lesion group(n=34)and a non-residual lesion group(n=86).Chi-square test and multivariate logistic regression analysis were performed to identify potential risk factors for residual lesions following the initial TURBT.Univariate and multivariate Cox regression models were used to analyze potential risk factors for tumor recurrence after the second TURBT.Results The residual lesion rate after initial TURBT was 28.33％.Chi-square test analysis revealed that tumor stage T1(Chi-square=5.756,P=0.016)and broad tumor base(Chi-square=4.331,P=0.037)were factors influencing residual lesions after initial TURBT.Multivariate logistic regression analysis identified tumor stage T1(OR=3.047,95％CI:1.128～8.226,P=0.028)as an independent risk factor for residual lesions after initial TURBT.The tumor recurrence rate after second TURBT was 17.5％.Multivariate Cox regression analysis identified tumor stage T1(OR=4.258,95％CI:1.248～14.532,P=0.021),intravesical chemotherapy instillation after second TURBT(OR=3.539,95％CI:1.284～9.752,P=0.015),history of urinary system tumors(OR=3.002,95％CI:1.145～7.873,P=0.025)and high platelet-to-lymphocyte(PLR)ratio(OR=2.798,95％CI:1.115～7.023,P=0.028)as independent risk factors for tumor recurrence after second TURBT.Conclusion Tumor stage T1 and broad tumor base are risk factors for residual lesions after initial TURBT,while tumor stage T1,intravesical chemotherapy instillation after second TURBT,history of urinary system tumors and high PLR ratio are risk factors for tumor recurrence after second TURBT.Comprehensive analysis on above 4 indicators can effectively assess the risk of tumor recurrence in NMIBC patients following second TURBT,and timely early medical intervention is beneficial for improving patient outcomes.

ObjectiveTo explore the effect of Shenge Bushen Capsules （SBC） on sexual development in normal 3-week-old mice. MethodsThe experiment consisted of two parts. In the first part， mice were divided into four groups： The control group and the low， medium， and high-dose SBC groups （234.7， 469.4， 938.7 mg·kg^-1， respectively）. In the second part， mice were divided into four groups： Control group， Pseudostellariae Radix polysaccharide （PRP） group， total flavonoids group， and SBC group， all receiving a dose of 469.4 mg·kg^-1. After 7 days of administration， the vaginal opening of female mice and the descent of testes and scrotum in male mice， as well as the ovarian and testicular organ indices， were observed. After 4 weeks of administration， female and male mice were housed together for 2 days， and the pregnancy rate of females was monitored. After delivery， the pregnant female mice continued receiving the treatment for 4 weeks， and the sexual development of their offspring， including vaginal opening， testicular descent， and organ indices of ovaries and testes， was observed. Serum sex hormones were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of gonadotropin-releasing hormone （GnRH） and growth hormone （GH） proteins in the hypothalamus was assessed by Western blot. ResultsCompared with the control group， there was no significant effect on the vaginal opening of female mice or the descent of testes in male mice after 7 days of SBC administration. After 4 weeks of administration， the pregnancy rate in the low-dose group was significantly reduced （P<0.05）， but no significant effects were observed in the other groups. The three doses of SBC did not significantly affect the ovarian or testicular organ indices， and there was no significant upregulation in the expression of GnRH or GH in the hypothalamus. The primary component of SBC， Pseudostellariae Radix polysaccharide， significantly reduced the vaginal opening in female mice after 7 days of administration （P<0.05）. After 4 weeks， the serum estradiol levels of non-pregnant female mice were decreased （P<0.05）， but there was no significant effect on the expression of GnRH or GH proteins in the hypothalamus of either male or female mice. Additionally， there were no significant effects on precocious puberty indicators， such as vaginal opening and testicular descent， in the offspring mice. ConclusionSBC does not significantly promote precocious puberty in young mice， and it does not have any noticeable effects on the pregnancy rate of adult mice or the sexual development of their offspring.

Objective:To investigate the role of a modified positioning device in improving image quality and diagnostic efficacy for thyroid cancer in contrast-enhanced neck CT imaging.Methods:This prospective cross-sectional study included 137 patients with pathologically confirmed thyroid lesions who underwent contrast-enhanced neck CT at Zhangzhou Affiliated Hospital of Fujian Medical University from January to April 2024. Patients scanned in January and February (modified positioning group, n=62) underwent scanning using the modified positioning device, whereas those scanned in March and April (traditional positioning group, n=75) underwent scanning with conventional positioning. The estimated volume CT dose index (CTDI vol) in the thyroid region was recorded. Subjective image quality for thyroid and neck regions was evaluated using a 5-point Likert scale. Diagnostic assessments for thyroid cancer, capsule invasion, and lymph node metastasis were independently conducted by one junior radiologist and one senior radiologist using a 5-point scoring system, with scores≥3 considered positive diagnoses. The differences of CTDI vol and image quality scores between the 2 groups were compared using Mann-Whitney U test. The diagnostic performance was evaluated by the receiver operating characteristic curve analysis. Results:The estimated CTDI vol values for the thyroid region were significantly lower in the modified positioning group compared to the traditional positioning group [11.20 (8.37, 13.56) vs. 12.46 (10.10, 19.43) mGy, Z=1.99, P=0.026]. Subjective image quality scores for thyroid and neck regions were significantly higher in the modified positioning group than in the traditional positioning group (all P<0.001). For thyroid cancer diagnosis by the senior radiologist, the modified positioning group had a significantly higher area under the curve (AUC) of 0.842 (95% CI 0.728-0.956) compared to the traditional positioning group (AUC=0.666,95% CI 0.554-0.777, Z=2.17, P=0.031). No significant differences were observed in diagnostic performance between the junior and senior radiologists for thyroid cancer, capsule invasion, and lymph node metastasis in other subgroup comparisons (all P>0.05). Conclusion:The modified positioning device using in contrast-enhanced neck CT imaging can improve image quality and diagnostic efficacy for thyroid cancer while reducing radiation exposure to the thyroid gland.

Objective:To investigate the role of a modified positioning device in improving image quality and diagnostic efficacy for thyroid cancer in contrast-enhanced neck CT imaging.Methods:This prospective cross-sectional study included 137 patients with pathologically confirmed thyroid lesions who underwent contrast-enhanced neck CT at Zhangzhou Affiliated Hospital of Fujian Medical University from January to April 2024. Patients scanned in January and February (modified positioning group, n=62) underwent scanning using the modified positioning device, whereas those scanned in March and April (traditional positioning group, n=75) underwent scanning with conventional positioning. The estimated volume CT dose index (CTDI vol) in the thyroid region was recorded. Subjective image quality for thyroid and neck regions was evaluated using a 5-point Likert scale. Diagnostic assessments for thyroid cancer, capsule invasion, and lymph node metastasis were independently conducted by one junior radiologist and one senior radiologist using a 5-point scoring system, with scores≥3 considered positive diagnoses. The differences of CTDI vol and image quality scores between the 2 groups were compared using Mann-Whitney U test. The diagnostic performance was evaluated by the receiver operating characteristic curve analysis. Results:The estimated CTDI vol values for the thyroid region were significantly lower in the modified positioning group compared to the traditional positioning group [11.20 (8.37, 13.56) vs. 12.46 (10.10, 19.43) mGy, Z=1.99, P=0.026]. Subjective image quality scores for thyroid and neck regions were significantly higher in the modified positioning group than in the traditional positioning group (all P<0.001). For thyroid cancer diagnosis by the senior radiologist, the modified positioning group had a significantly higher area under the curve (AUC) of 0.842 (95% CI 0.728-0.956) compared to the traditional positioning group (AUC=0.666,95% CI 0.554-0.777, Z=2.17, P=0.031). No significant differences were observed in diagnostic performance between the junior and senior radiologists for thyroid cancer, capsule invasion, and lymph node metastasis in other subgroup comparisons (all P>0.05). Conclusion:The modified positioning device using in contrast-enhanced neck CT imaging can improve image quality and diagnostic efficacy for thyroid cancer while reducing radiation exposure to the thyroid gland.

Objective This study aims to investigate the impact of cultivation time on dendritic cells(DCs)and their derived exosomes′ expression of immune-related membrane proteins(CD80,MHC-Ⅰ,MHC-Ⅱ)and provides experimental evidence for future research.Methods Mouse bone marrow cells were induced to differentiate into DCs using GM-CSF and IL-4,followed by maturation stimulation withTNF-α.Exosomes were extracted using ultracentrifugation.Western blot and Amnis image flow cytometry were used to identify exosomes derived from mouse DCs.Amnis image flow cytometry was used to detect the expression of immune-related proteins CD80,CD11c,MHC-Ⅰ,and MHC-Ⅱ in mouse DCs and their exosomes.Results After 5 days of in vitro cultivation,more than 50%of dendritic cells expressed CD80,CD11c,MHC-Ⅰ,and MHC-Ⅱ,reaching the highest level on day 13.The positivity rates were as follows:CD80(97.29±0.63)%,CD11c(92.31±1.18)%,MHC-Ⅰ(97.91±0.49)%,and MHC-Ⅱ(97.91±0.49)%.The differences were statistically significant(P<0.001).The expression gradually decreased after day 13,but approximately 80%of DC cells still expressed MHC-Ⅰ and MHC-Ⅱ immune molecules on day 30.The expression levels of CD80,CD11c,and MHC-Ⅱ on the exosome membrane were highest on day 5 and then decreased overall with prolonged cultivation time,except for MHC-Ⅰ molecules.The differences were statistically significant(P<0.01).Conclusions In vitro-cultured mouse dendritic cells express high levels of immune-related membrane proteins and can be stably maintained for a long time under suitable culture conditions.The secreted exosomes also carry abundant immune-related membrane proteins,but no significant correlation was found between the immune-related proteins on the dendritic cell surface and the exosome membrane surface.