Objective: To investigate the prevalence of depressive symptoms and their associated factors among students in Zhejiang Province, so as to provide evidence for targeted prevention strategies. Methods: A stratified cluster random sampling method was used to select 23 829 college students and primary and secondary school students aged 11-22 years in Zhejiang Province from December 2019 to February 2020. Depressive symptoms were assessed using the Center for Epidemiologic Studies Depression Scale (CES-D). Three machine learning algorithms, including Logistic regression, random forest, and eXtreme Gradient Boosting (XGBoost), were applied to construct predictive models, and key associated factors were identified by comparing model performance. Results: The detection rate of depressive symptoms among students in Zhejiang Province was 19.92%; the rates were 17.20% in boys and 22.87% in girls( χ 2=164.89, P <0.05). The CES-D total score was 9.00(4.00,13.00). Multiple Logistic regression analysis revealed that loneliness had the strongest association with depressive symptoms ( AOR =9.58, 95% CI =8.90-10.30), while bullying exposure ( AOR =4.39, 95% CI =4.02-4.80), female students( AOR =1.81, 95% CI =1.68-1.94)，never eating breakfast ( AOR = 2.34，95% CI =2.00-2.67) and overweight/obesity( AOR =1.10，95% CI =1.08-1.12) were significant associated factors of depressive symptoms among students (all P <0.05). Analysis based on the XGBoost model produced highly consistent results, identifying the above 5 factors as the core features with the highest correlation strength (all P <0.05). Conclusions Female, loneliness, bullying exposure, frequency of weekly breakfast and BMI are strongly associated with depressive symptoms among students. Mental health education for high risk groups should be strengthened, and coordinated prevention efforts between families and schools are recommended.

Objective:To evaluate the safety and efficacy of secondary transport on extracorporeal membrane oxygenation (ECMO) for critically ill children.Methods:This was a retrospective cohort study. Data from 222 pediatric patients who underwent ECMO transport from May 2019 to May 2024 at 5 ECMO centers and Chinese Database of Pediatric Extracorporeal Life Support Organization were collected. The cases were divided into primary and secondary transport groups by nature of transport. The clinical data, including demographics, ECMO indications, transport distance, pre-transport lab results, prognosis and complications were analyzed. Two independent samples t-test, Wilcoxon test, and χ2 test or Fisher′s exact probability method were used to compare the differences between 2 groups and evaluate the safety and efficacy of secondary transport. Results:Among the 222 children transported with ECMO, there were 135 males and 87 females, with an age of 3.0 (0.2, 7.0) years. There were 202 cases in the primary transport group and 20 cases in the secondary transport group. All secondary transport patients had failed attempts at weaning ECMO before transfer. The patients in the secondary transport group were older, had higher rates of surgical cannulation, circulatory support, and pre-ECMO lactate levels compared to the primary transport group (7.0 (2.8, 10.0) vs. 3.0 (0.2, 6.0) years old, 55.0% (11/20) vs. 3.6% (7/202), 80.0% (16/20) vs. 41.6% (84/202), (10±4) vs. (7±6) mmol/L, Z=3.41, χ 2=66.31, 10.99, t=2.24, all P<0.05). In the secondary transport group, the vasoactive-inotropic scores of patients on circulatory support and the oxygenation index for patients requiring respiratory support were higher than those in the primary transport group (83±33 vs. 82±68, 51.0±1.8 vs. 37.4±10.2, t=2.36, 2.63, respectively; both P<0.05). There were no statistically significant differences between the 2 groups in sex, transport distance, pre-ECMO creatinine, arterial blood gas BE values, and ECMO duration (all P>0.05). No life-threatening complications occurred during the transport in either group. Two patients in the secondary transport group underwent heart transplantation, and 1 patient underwent radiofrequency ablation. The overall survival rate between the 2 groups showed no statistically significant difference (45.0% (9/20) vs. 55.4% (112/202), χ2=1.15, P>0.05). Conclusions:Secondary ECMO transport for critically ill children don't increase mortality or life-threatening complications during transport. ECMO patients who cannot receive effective treatment locally can benefit from secondary transport to an advanced ECMO center provides further treatment opportunities.

BACKGROUND:Restoring the normal level of reactive oxygen species and mitochondrial function of nucleus pulposus cells and inhibiting apoptosis of nucleus pulposus cells are key targets for delaying intervertebral disc degeneration.Prussian blue nanoparticles have peroxidase-like activity,which can effectively remove reactive oxygen species in the pathological microenvironment and protect nucleus pulposus cells from oxidative stress damage.OBJECTIVE:To investigate the biological functions and mechanisms of Prussian blue nanoparticles in delaying nucleus pulposus degeneration in rats.METHODS:Prussian blue nanoparticles were prepared by hydrothermal method,and their micromorphology and particle size were characterized.Prussian blue nanoparticles with different mass concentrations(20,40,60,80,and 100 μg/mL)were used to intervene in the caudal nucleus pulposus cells of passage 2 SD rats.Cell proliferation was detected by CCK-8 assay after 24 hours.60 μg/mL Prussian blue nanoparticles were used to intervene in the caudal nucleus pulposus cells of passage 2 SD rats.The viability of nucleus pulposus cells was observed by live-dead staining after 1 and 3 days.Passage 2 SD rat caudal vertebrae nucleus pulposus cells were obtained and observed for cell adhesion before being divided into three intervention groups.The control group did not receive any intervention.The lipopolysaccharide group was added with lipopolysaccharide.The lipopolysaccharide+Prussian blue nanoparticle group was added with lipopolysaccharide and 60 μg/mL Prussian blue nanoparticles.Reactive oxygen species,mitochondrial superoxide,and mitochondrial membrane potential were detected 24 hours after intervention.RT-qPCR detection and Alcian blue staining were performed 48 hours after intervention.RESULTS AND CONCLUSION:(1)Under transmission electron microscopy,Prussian blue nanoparticles were uniform nanocubes with an average particle size of 130 nm.(2)CCK-8 assay results showed that 20-60 μg/mL Prussian blue nanoparticles had no obvious cytotoxicity,and 60 μg/mL Prussian blue nanoparticles were selected for cell intervention in subsequent experiments.Live-dead staining results showed that 60 μg/mL Prussian blue nanoparticles had no effect on the viability of nucleus pulposus cells.(3)Compared with the control group,the levels of reactive oxygen species and mitochondrial superoxide in nucleus pulposus cells in the lipopolysaccharide group were increased(P＜0.01),the mitochondrial membrane potential was decreased(P＜0.01),the mRNA expressions of type Ⅱ collagen and aggrecan were decreased(P＜0.01),the mRNA expressions of matrix metalloproteinase 13 and thrombospondin integrin metallopeptidase 5 were increased(P＜0.01),and the positive area of Alcian blue staining was reduced(P＜0.01).Compared with the lipopolysaccharide group,the levels of reactive oxygen species and mitochondrial superoxide in nucleus pulposus cells in the lipopolysaccharide+Prussian blue nanoparticle group were decreased(P＜0.01),mitochondrial membrane potential increased(P＜0.01),mRNA expression of type Ⅱ collagen and aggrecan increased(P＜0.01),mRNA expression of matrix metalloproteinase 13 and thrombospondin integrin metallopeptidase 5 decreased(P＜0.01),and positive area of Alcian blue staining increased(P＜0.01).The results showed that Prussian blue nanoparticles delayed the degeneration of rat nucleus pulposus by reducing oxidative stress of nucleus pulposus cells,restoring mitochondrial function,and maintaining the balance of extracellular matrix synthesis and catabolism.

Objective·To explore the antitumor effects and potential mechanisms of combining phosphoinositide 3-kinase,FYVE-type zinc finger containing(PIKfyve)inhibitor YM201636 with the stimulator of interferon genes(STING)agonist diABZI in STING pathway-deficient melanoma.Methods·The mRNA and protein expression levels of STING in human cancer cell lines were obtained from the Cancer Cell Line Encyclopedia(CCLE)and UniProt databases.Based on median expression values,melanoma cell lines with high STING mRNA but low protein expression were identified.Quantitative real-time PCR(qRT-PCR)and Western blotting were performed to validate STING mRNA and protein expression in human melanoma cells.The murine melanoma cell line YUMM1.7,characterized by low STING protein expression,was selected through Western blotting.The ability of YM201636 to restore STING protein expression in YUMM1.7 cells was evaluated.STING agonist diABZI was then applied in combination with YM201636 to analyze the synergistic tumor cell-killing effect through CCK-8 assay.Western blotting was used to detect the phosphorylation of TANK-binding kinase 1(TBK1)and interferon regulatory factor 3(IRF3),and qRT-PCR was used to evaluate type Ⅰ interferon expression.A mouse melanoma model was established and treated with YM201636,diABZI,or their combination.Tumor volume was measured,and treatment efficacy was assessed.RNA sequencing and immunofluorescence staining were performed to analyze immune cell infiltration in the tumor microenvironment.Results·Database analyses,qRT-PCR,and Western blotting confirmed that some human melanoma cell lines exhibited high STING mRNA expression but low STING protein levels.YM201636 significantly increased STING protein expression in YUMM1.7 cells(P<0.001).Combined treatment with YM201636 and diABZI significantly enhanced phosphorylation of TBK1 and IRF3(P<0.05),indicating effective activation of the STING signaling pathway.This combination also promoted the expression of type Ⅰ interferons(P<0.001)and enhanced tumor cell killing in vitro.In vivo,the combination therapy markedly suppressed melanoma growth compared to monotherapy.Immune profiling of the tumor microenvironment revealed significantly increased infiltration of CD4? T cells and CD8? T cells in the combination treatment group(P<0.05).Conclusion·The PIKfyve inhibitor YM201636 could restore STING protein expression in STING-deficient melanoma and enhance the antitumor efficacy of the STING agonist diABZI,offering a promising therapeutic strategy for tumors with defective STING signaling.

Objective:To investigate the semen characteristics of male patients with autosomal dominant polycystic kidney disease (ADPKD) complicated by infertility and the impact of PKD1 mutations on their reproductive outcomes following preimplantation genetic testing for monogenic diseases (PGT-M). Methods:A retrospective cohort study was conducted to analyze the clinical data of ADPKD patients in the Clinical Center of Reproductive Medicine, the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2023. The study included two groups: observation group consisting of male infertility patients with ADPKD aged 20-45 years ( n=78), and control group comprising ADPKD patients with PKD1 mutations who demonstrated normal semen parameters ( n=5). According to the type of mutation, the patients were further divided into the PKD1 truncated mutation group and the non-truncated mutation group. Baseline data, ovulation induction outcomes and PGT-M clinical outcomes were compared between the two groups. Binary logistic regression analysis was used to assess the effects of age, family history, mutation type and mutation position on semen quality and live birth rates. Results:In the observation group, there were 28 cases of asthenozoospermia, 38 cases of oligoasthenozoospermia, and 12 cases of azoospermia. The total sperm count and forward motility ratio of patients with asthenozoospermia, oligoasthenozoospermia, and azoospermia in the observation group were significantly different from those in control group (all P<0.001). Genetic diagnosis revealed PKD1 mutations in 70 cases, PKD2 mutation in 1 case, and no known pathogenic mutations were identified in 7 cases. Among the 70 ADPKD patients with PKD1 mutations, 52 couples underwent PGT-M assisted reproduction. The blastocyst formation rate was significantly lower in the non-truncating mutation group [44.74% (51/114)] than in the truncating mutation group [58.76% (161/274), P=0.011]. No statistically significant differences were observed between the two groups in other parameters including age of both partners, semen parameters, number of oocytes retrieved, two pronuclei (2PN) rate, number of available embryos, high-quality embryo rate, clinical pregnancy rate, miscarriage rate, and live birth rate (all P>0.05). Binary logistic regression analysis showed that neither PKD1 mutation type nor PKD1 mutation site was an independent factor affecting semen parameters and live birth rate in ADPKD patients (all P>0.05). Conclusions:Asthenospermia and oligoasthenospermia are the most common semen phenotypes in ADPKD-associated male infertility. The PKD1 mutation type and location are not associated with abnormal semen parameters in ADPKD-associated infertility patients. PKD1 mutation types do not affect the outcomes of PGT-M in infertile patients with ADPKD.

Objective To explore the association between single nucleotide polymorphism(SNP)of Rab37 gene and the age at menarche(AAM).Methods A case-control study design was used to conduct an epidemiologic survey on 2 060 women in a community in Jurong City,Jiangsu Province,and their venous blood samples were collected.ASA chip was used to explore the Rab37 gene genotyping of blood sample DNA to obtain the genotyping results for Rab37 SNPs rs62084865,rs10512597,rs35489971,rs1037170,rs6501732,rs77822106,and rs3178300.Based on menarche time of＜16 years old or≥16 years old,the participants were divided into 2 groups.Linear and logistic regression analyses were used to explore the association between the SNP of Rab37 gene and the age at menarche based on dominant,recessive and allelic models.Results Among the 2 060 healthy women,944 women had normal menstrual periods,while 1 116 women experienced delayed menarche.Linear regression analysis showed that the gene polymorphism of rs3178300 was negatively correlated with the age at menarche in females[recessive model:CC/CT+TT,β=-0.915,95%CI(-1.692,-0.137),P=0.021;allelic model:T/C,β=-0.221,95%CI(-0.410,-0.032),P=0.022].Logistic regression analysis indicated that the gene polymorphism of rs3178300 was significantly associated with the risk of delayed menarche in females[recessive model:CC/CT+TT,OR=0.295,95%CI(0.116,0.751),P=0.010;allelic model:T/C,OR=0.796,95%CI(0.652,0.972),P=0.025].The remaining 6 SNPs showed no significant association with age at menarche.Conclusion The rs3178300 polymorphism of Rab37 gene is significantly associated with delayed menarche in Chinese women.

The p21-activated kinase 4 (PAK4), a key regulator of malignancy, is negatively correlated with immune infiltration and has become an emergent drug target of cancer therapy. Given the lack of high efficacy PAK4 inhibitors, we herein reported the identification of a novel inhibitor 13 bearing a tetrahydrobenzofuro[2,3-c]pyridine tricyclic core and possessing high potency against MIA PaCa-2 and Pan02 cell lines with IC₅₀ values of 0.38 and 0.50 μmol/L, respectively. This compound directly binds to PAK4 in a non-ATP competitive manner. In the mouse Pan02 model, compound 13 exhibited significant tumor growth inhibition at a dose of 100 mg/kg, accompanied by reduced levels of PAK4 and its phosphorylation together with immune infiltration in mice tumor tissue. Overall, compound 13 is a novel allosteric PAK4 inhibitor with a unique tricyclic structural feature and high potency both in vitro and in vivo, thus making it worthy of further exploration.

OBJECTIVES: To analyze the differences in the prognosis of gastric signet ring cell carcinoma (SRCC) among different races using the US Surveillance Epidemiology and End Results (SEER) database and The Cancer Genome Atlas (TCGA) database. METHODS: We analyzed the data of patients with gastric SRCC from the SEER database from 2000 to 2020, and divided the patients into cohorts of whites, blacks, Asians or Pacific Islanders, American Indians/Alaska Natives according to their race. The prognosis and treatment of the cohorts were evaluated using baseline demographic analysis, Kamplan-Meier survival curve, and nomogram analysis. RESULTS: We analyzed the data of a total of 2058 patients, including 8.6% blacks, 72.4% whites, 16.6% Asians or Pacific Islanders, 1.0% American Indians/Alaska Natives, and 1.4% other races. The tumor grade varied among different races, and the prevalence and survival rates of patients differed significantly across races. The differences in the white cohort were the most prominent, and all the differences were statistically significant (P<0.05). Racial differences were also noted in patient management and prognosis. CONCLUSIONS There are racial differences in tumor grades and prognosis of gastric SRCC, and these differences provide evidence for optimizing clinical diagnosis and treatment strategies for this malignancy.
Aged ; Female ; Humans ; Male ; Middle Aged ; Carcinoma, Signet Ring Cell/therapy* ; Databases, Factual ; Prognosis ; Racial Groups ; SEER Program ; Stomach Neoplasms/therapy* ; Survival Rate ; United States/epidemiology* ; White ; Asian American Native Hawaiian and Pacific Islander ; American Indian or Alaska Native ; Black or African American

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

BACKGROUND:Restoring the normal level of reactive oxygen species and mitochondrial function of nucleus pulposus cells and inhibiting apoptosis of nucleus pulposus cells are key targets for delaying intervertebral disc degeneration.Prussian blue nanoparticles have peroxidase-like activity,which can effectively remove reactive oxygen species in the pathological microenvironment and protect nucleus pulposus cells from oxidative stress damage.OBJECTIVE:To investigate the biological functions and mechanisms of Prussian blue nanoparticles in delaying nucleus pulposus degeneration in rats.METHODS:Prussian blue nanoparticles were prepared by hydrothermal method,and their micromorphology and particle size were characterized.Prussian blue nanoparticles with different mass concentrations(20,40,60,80,and 100 μg/mL)were used to intervene in the caudal nucleus pulposus cells of passage 2 SD rats.Cell proliferation was detected by CCK-8 assay after 24 hours.60 μg/mL Prussian blue nanoparticles were used to intervene in the caudal nucleus pulposus cells of passage 2 SD rats.The viability of nucleus pulposus cells was observed by live-dead staining after 1 and 3 days.Passage 2 SD rat caudal vertebrae nucleus pulposus cells were obtained and observed for cell adhesion before being divided into three intervention groups.The control group did not receive any intervention.The lipopolysaccharide group was added with lipopolysaccharide.The lipopolysaccharide+Prussian blue nanoparticle group was added with lipopolysaccharide and 60 μg/mL Prussian blue nanoparticles.Reactive oxygen species,mitochondrial superoxide,and mitochondrial membrane potential were detected 24 hours after intervention.RT-qPCR detection and Alcian blue staining were performed 48 hours after intervention.RESULTS AND CONCLUSION:(1)Under transmission electron microscopy,Prussian blue nanoparticles were uniform nanocubes with an average particle size of 130 nm.(2)CCK-8 assay results showed that 20-60 μg/mL Prussian blue nanoparticles had no obvious cytotoxicity,and 60 μg/mL Prussian blue nanoparticles were selected for cell intervention in subsequent experiments.Live-dead staining results showed that 60 μg/mL Prussian blue nanoparticles had no effect on the viability of nucleus pulposus cells.(3)Compared with the control group,the levels of reactive oxygen species and mitochondrial superoxide in nucleus pulposus cells in the lipopolysaccharide group were increased(P＜0.01),the mitochondrial membrane potential was decreased(P＜0.01),the mRNA expressions of type Ⅱ collagen and aggrecan were decreased(P＜0.01),the mRNA expressions of matrix metalloproteinase 13 and thrombospondin integrin metallopeptidase 5 were increased(P＜0.01),and the positive area of Alcian blue staining was reduced(P＜0.01).Compared with the lipopolysaccharide group,the levels of reactive oxygen species and mitochondrial superoxide in nucleus pulposus cells in the lipopolysaccharide+Prussian blue nanoparticle group were decreased(P＜0.01),mitochondrial membrane potential increased(P＜0.01),mRNA expression of type Ⅱ collagen and aggrecan increased(P＜0.01),mRNA expression of matrix metalloproteinase 13 and thrombospondin integrin metallopeptidase 5 decreased(P＜0.01),and positive area of Alcian blue staining increased(P＜0.01).The results showed that Prussian blue nanoparticles delayed the degeneration of rat nucleus pulposus by reducing oxidative stress of nucleus pulposus cells,restoring mitochondrial function,and maintaining the balance of extracellular matrix synthesis and catabolism.