Objective: To understand the impact of childhood trauma on psychological distress among upper grade elemetary school students, and to explore the mediating role of leisure activities in the relationship, so as to provide a basis for developing mental health intervention strategies. Methods: From August to November 2024, a combination of convenience sampling and stratified cluster random sampling was employed to recruit 1 373 fourth to sixth grade students from four primary schools in Harbin. The Childhood Trauma Questionnaire(CTQ), a self designed leisure activity scale (including active and passive leisure activities), and the Kessler Psychological Distress Scale (K10) were used to assess childhood trauma experiences, leisure activities, and levels of psychological distress. Spearman correlation analysis and linear regression analysis were conducted to explore the relationships among childhood trauma, leisure types, leisure time, and psychological distress. Based on the mediation analysis framework proposed by Hayes (Model 4), the mediating role of leisure types in the relationship between childhood trauma and psychological distress was examined. Results: Totally 19.1% of the upper elemetary school students exhibited psychological distress, while 30.2% had experienced childhood trauma. During school days, 64.6% of the students were reported of having leisure time concentrated between 1 and 5 hours per day, whereas 67.4% reported leisure time exceeding 5 hours per day on weekends. After controlling for potential demographic confounders such as gender, grade, ethnicity, household registration, being an only child, parents educational level, co residence, and whether parents are first time married,linear regression analysis showed that childhood trauma experience had positive predictive effect on psychological distress in upper primary school students( β =0.20, P <0.01). Leisure time showed no statistically significant association with psychological distress, both on school days ( β =-0.58 to -0.56) and weekends ( β =0.26- 0.98 )(all P >0.05). Active leisure activities were negatively associated with psychological distress ( β =-0.20), while passive leisure activities were positively associated with psychological distress ( β =0.29)(both P <0.01). Leisure type partially mediated the relationship between childhood trauma and psychological distress, accounting for 11.7% of the indirect effect. Conclusion Childhood trauma experiences positively predict psychological distress in upper elementary school students, and affect psychological distress through active leisure and passive leisure.

Objective This study aims to explore the efficacy and complication rates of the transcranial approach(TCA)and extended endoscopic endonasal approach(EEEA)for the treatment of craniopharyngiomas based on the QST classification,providing a scientific reference for clinical decision-making on surgical approach.Methods A total of 151 patients who underwent craniopharyngioma surgery at our center from January 2018 to December 2023 were enrolled.The patients were categorized into Q-CP(suprasellar type),S-CP(infundibular type),and T-CP(tuberal type)according to the QST classification.Systematic collection and analysis were performed on the outcomes of TCA and EEEA treatments,respectively.The differences in effectiveness between the two approaches were evaluated based on the QST classification.Results The improvement rate of visual symptoms was overall higher in the EEEA group than in the TCA group(59.1%vs.36.5%,P=0.006),and the visual deterioration rate was lower(3.0%vs.14.1%,P=0.006).However,the incidence of cerebrospinal fluid leakage was higher in the EEEA group(15.2%vs.3.5%,χ2=4.986,P=0.026).The incidence of postoperative seizures(8.2%vs.0,P=0.019),brain contusions(10.6%vs.0,P=0.005),and subdural hematoma(9.4%vs.0%,P=0.01)was higher in the TCA group.For patients with Q-CP type,the EEEA group had a higher rate of total tumor resection(92.9%vs.65.2%,P=0.025)and a lower recurrence rate(3.6%vs.21.7%,P=0.047),with shorter hospital stays and lower postoperative costs.The TCA group had higher intraoperative blood loss in this type(300 mL vs.200 mL,Z=-2.261,P=0.024).For S-CP type patients,the EEEA group showed a higher rate of total tumor resection(91.3%vs.74.2%)and a lower recurrence rate(0 vs.12.9%,P=0.031),with lower postoperative hospital costs.In T-CP type,due to the deeper location,EEEA showed limitations in protecting hypothalamic function and the TCA group had a better postoperative hypothalamic function score(P=0.035).Conclusion Based on QST classification,EEEA has advantages in Q-CP and S-CP types and is recommended as the preferred surgical procedure;In the T-CP type,TCA surgery is more helpful in protecting hypothalamic function.

Objective: To explore the impact of sleep quality, experience of childhood maltreatment, and their interaction on depressive symptoms among middle school students, so as to provide the reference for early intervention of depressive symptoms among middle school students. Methods: From September to December 2023, a questionnaire survey was conducted among 1 231 students from two secondary schools in Harbin, Heilongjiang Province by a convenient sampling method. The survey included general demographic information, Childhood Trauma Questionnaire Short Form, Pittsburgh Sleep Quality Index and Short Version of Center for Epidemiological Studies Depression Scale. The Chi square test was used to analyze the differences in depressive symptom, sleep quality and childhood maltreatment among students with different demographic characteristics. Correlation analysis was conducted using Logistic regression, and interaction analysis was performed by both additive and multiplicative interaction models. Results: The detection rate of depressive symptoms among middle school students was 22.7%, and the rate for high school students (35.2%) was significantly higher than that for middle school students (17.0%) ( χ 2=50.35, P <0.01). The detection rates of depressive symptoms among middle school students with a history of childhood maltreatment and poor sleep quality were 45.8% and 44.0%, respectively. Multivariate Logistic regression analysis showed that compared to students without a history of childhood maltreatment, students with a history of childhood maltreatment had a higher risk of depressive symptoms ( OR =4.49，95% CI =3.31~ 6.09 , P <0.01);students with poor sleep quality had a higher risk of depressive symptoms than students with good sleep quality ( OR = 5.99，95% CI =4.37~8.22, P <0.01).The interaction results showed that the presence of childhood maltreatment and poor sleep quality had an additive interaction on the occurrence of depression in middle school students. Compared with students without childhood maltreatment and having good sleep quality, students with childhood maltreatment and poor sleep quality had a 22.49 times higher risk of developing depression ( OR =22.49，95% CI =14.22~35.59, P <0.01). Conclusion Depressive symptoms among middle school students are associated with childhood maltreatment and poor sleep quality, and there is an additive interaction between childhood maltreatment and poor sleep quality on the impact of depressive symptoms.

Objective:To study the effect of adipose tissue derived extracellular vesicles (AT-EVs) on nerve regeneration and functional reinnervation after sciatic nerve injury (SNI) in rat.Methods:From April 2023 to April 2024, the experiment was completed by the Department of Orthopaedics, the First Affiliated Hospital of Zhengzhou University in the key laboratory of the School of Medical Sciences, Zhengzhou University. AT-EVs were isolated and extracted from subcutaneous adipose tissue of 12 healthy male SD rats using ultracentrifugation. The morphology, size and distribution of AT-EVs were characterised via transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The effect of AT-EVs on the proliferation ability of pheochromocytoma cells of rat (PC12) and rat Schwann cells (RSC 96) were detected by cell proliferation assay. Twenty-four healthy male SD rats were randomly divided into 3 groups with 8 rats per group, including a sham group, a SNI group and an AT-EVs treatment group. Right sciatic nerves of rat models in sham group were exposed without injury. For the rat models in SNI group and AT-EVs treatment group, right sciatic nerves were injected with phosphate buffered solution (PBS) and AT-EVs under the epineurium of the nerve respectively following sciatic nerve crush. At 28 days after surgery, Sciatic function index (SFI) and pace analysis was performed. Then, rats in all groups were sacrificed, and bilateral gastrocnemius muscles were collected to measure the muscle wet weight ratio. The right sciatic nerves were removed for further exploration of nerve regeneration. The histopathological changes and axon arrangement of sciatic nerve were evaluated by Hematein Eosin (HE) staining and Masson staining. The regeneration efficiency of Myelin forming protein (MBP) and Neuronal marker [β-tubulin (Tuj-1)] were analysed by immunofluorescence staining of sciatic nerve. Statistical analysis was conducted using GraphPad Prism 8.0. P<0.05 were considered statistically significant. Results:The results of TEM and NTA revealed that AT-EVs exhibited a typical disc-shaped bilayer membrane structure with a predominant particle size of 184.5 nm. AT-EVs significantly enhanced the proliferation ability of PC12 and RSC96, and the difference was statistically significant ( P<0.05). At 28 days after surgery, the SFI and muscle wet weight ratio were higher in the AT-EVs treatment group compared to the SNI group, and the difference was statistically significant ( P<0.05). Axons in AT-EVs treatment group were arranged more closely and orderly than those in SNI group at 28 days after surgery, and there were less axon disintegration and vacuolation caused by injury. The immunofluorescence assay indicated that the MBP and Tuj-1 fluorescence intensity in AT-EVs treatment group was significantly higher than those in the SNI group, and the difference was statistically significant ( P<0.05). Conclusion:AT-EVs can enhance the proliferation ability of PC12 and RSC96 to promote axon regeneration and functional reinnervation following SNI.

Objective This study aims to explore the efficacy and complication rates of the transcranial approach(TCA)and extended endoscopic endonasal approach(EEEA)for the treatment of craniopharyngiomas based on the QST classification,providing a scientific reference for clinical decision-making on surgical approach.Methods A total of 151 patients who underwent craniopharyngioma surgery at our center from January 2018 to December 2023 were enrolled.The patients were categorized into Q-CP(suprasellar type),S-CP(infundibular type),and T-CP(tuberal type)according to the QST classification.Systematic collection and analysis were performed on the outcomes of TCA and EEEA treatments,respectively.The differences in effectiveness between the two approaches were evaluated based on the QST classification.Results The improvement rate of visual symptoms was overall higher in the EEEA group than in the TCA group(59.1%vs.36.5%,P=0.006),and the visual deterioration rate was lower(3.0%vs.14.1%,P=0.006).However,the incidence of cerebrospinal fluid leakage was higher in the EEEA group(15.2%vs.3.5%,χ2=4.986,P=0.026).The incidence of postoperative seizures(8.2%vs.0,P=0.019),brain contusions(10.6%vs.0,P=0.005),and subdural hematoma(9.4%vs.0%,P=0.01)was higher in the TCA group.For patients with Q-CP type,the EEEA group had a higher rate of total tumor resection(92.9%vs.65.2%,P=0.025)and a lower recurrence rate(3.6%vs.21.7%,P=0.047),with shorter hospital stays and lower postoperative costs.The TCA group had higher intraoperative blood loss in this type(300 mL vs.200 mL,Z=-2.261,P=0.024).For S-CP type patients,the EEEA group showed a higher rate of total tumor resection(91.3%vs.74.2%)and a lower recurrence rate(0 vs.12.9%,P=0.031),with lower postoperative hospital costs.In T-CP type,due to the deeper location,EEEA showed limitations in protecting hypothalamic function and the TCA group had a better postoperative hypothalamic function score(P=0.035).Conclusion Based on QST classification,EEEA has advantages in Q-CP and S-CP types and is recommended as the preferred surgical procedure;In the T-CP type,TCA surgery is more helpful in protecting hypothalamic function.

Objective:To study the effect of adipose tissue derived extracellular vesicles (AT-EVs) on nerve regeneration and functional reinnervation after sciatic nerve injury (SNI) in rat.Methods:From April 2023 to April 2024, the experiment was completed by the Department of Orthopaedics, the First Affiliated Hospital of Zhengzhou University in the key laboratory of the School of Medical Sciences, Zhengzhou University. AT-EVs were isolated and extracted from subcutaneous adipose tissue of 12 healthy male SD rats using ultracentrifugation. The morphology, size and distribution of AT-EVs were characterised via transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The effect of AT-EVs on the proliferation ability of pheochromocytoma cells of rat (PC12) and rat Schwann cells (RSC 96) were detected by cell proliferation assay. Twenty-four healthy male SD rats were randomly divided into 3 groups with 8 rats per group, including a sham group, a SNI group and an AT-EVs treatment group. Right sciatic nerves of rat models in sham group were exposed without injury. For the rat models in SNI group and AT-EVs treatment group, right sciatic nerves were injected with phosphate buffered solution (PBS) and AT-EVs under the epineurium of the nerve respectively following sciatic nerve crush. At 28 days after surgery, Sciatic function index (SFI) and pace analysis was performed. Then, rats in all groups were sacrificed, and bilateral gastrocnemius muscles were collected to measure the muscle wet weight ratio. The right sciatic nerves were removed for further exploration of nerve regeneration. The histopathological changes and axon arrangement of sciatic nerve were evaluated by Hematein Eosin (HE) staining and Masson staining. The regeneration efficiency of Myelin forming protein (MBP) and Neuronal marker [β-tubulin (Tuj-1)] were analysed by immunofluorescence staining of sciatic nerve. Statistical analysis was conducted using GraphPad Prism 8.0. P<0.05 were considered statistically significant. Results:The results of TEM and NTA revealed that AT-EVs exhibited a typical disc-shaped bilayer membrane structure with a predominant particle size of 184.5 nm. AT-EVs significantly enhanced the proliferation ability of PC12 and RSC96, and the difference was statistically significant ( P<0.05). At 28 days after surgery, the SFI and muscle wet weight ratio were higher in the AT-EVs treatment group compared to the SNI group, and the difference was statistically significant ( P<0.05). Axons in AT-EVs treatment group were arranged more closely and orderly than those in SNI group at 28 days after surgery, and there were less axon disintegration and vacuolation caused by injury. The immunofluorescence assay indicated that the MBP and Tuj-1 fluorescence intensity in AT-EVs treatment group was significantly higher than those in the SNI group, and the difference was statistically significant ( P<0.05). Conclusion:AT-EVs can enhance the proliferation ability of PC12 and RSC96 to promote axon regeneration and functional reinnervation following SNI.

Objective:To investigate the species and number of glial cell clusters, main signaling pathway and progression of glial cell clusters after peripheral nerve injury (PNI) in rats using single-cell RNA sequencing (scRNA-seq).Methods:Twenty-seven SD rats were randomly divided into sham-operated group, group of 3 d after PNI and group of 7 d after PNI ( n=9). Thrice squeezing the right sciatic nerves in the group of 3 d after PNI and group of 7 d after PNI and no damage in the sham-operated group were performed. Species and number of glial cell clusters in the right sciatic nerve samples were detected by scRNA-seq. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze the signal pathway enriched by differentially expressed genes (DEGs) of glial cells in the right sciatic nerve samples. Pseudo time analysis was used to simulate the progression of glial cell clusters in the right sciatic nerve samples. Results:(1) ScRNA-seq revealed a total of 1 609 glial cells (mainly cluster 6 [ n=1 388]) in the sciatic nerve samples of sham-operated group; 6 176 glial cells were observed in the sciatic nerve samples of group of 3 d after PNI, mainly cluster 2 ( n=3 124) and cluster 3 ( n=959); 8 975 glial cells were observed in the sciatic nerve samples of group of 7 d after PNI, mainly cluster 1 ( n=3 071), cluster 4 ( n=1 696), and cluster 5 ( n=1 389). (2) GO and KEGG analysis showed that compared with those in the sham-operated group, biological processes such as protein translation, cadherin binding and ribosome composition were up-regulated in glial cells in the group of 3 d after PNI and group of 7 d after PNI. Compared with those in the group of 3 d after PNI, glial cells enriched in biological processes such as axonal regeneration, myelination and focal adhesion, and in upregulated mitogen activated protein kinase (MAPK) signaling pathway and phosphoinositide 3-kinase (PI3K)-protein kinase B (PKB) signaling pathway in the group of 7 d after PNI. (3) Pseudo time analysis showed that glial cells in the sciatic nerve samples were mainly cluster 6 (marker genes: Atp1a2 and Sparcl1) in the sham-operated group, progressed into cluster 2 (marker genes: Mapt and Slc7a11) and cluster 3 (marker genes: Esco2 and Neil3) mainly in the group of 3 d after PNI, and progressed into cluster 1 (marker genes: Bcas1 and Prx), cluster 4 (marker genes: Ccn2 and Gap43), and cluster 5 (marker genes: Cd24 and Atxn1) mainly in the group of 7 d after PNI. Conclusion:In rats after PNI, glial cells can up-regulate MAPK signaling pathway and PI3K-PKB signaling pathway; with prolonged injury time, glial cells can progress into clusters with marker genes Bcas1, Prx, Cd24 and Atxn1 mainly.

Objective:To reveal the molecular characteristics of mononuclear phagocytes (MPs) in rat model of peripheral nerve injury (PNI) using single-cell RNA sequencing (scRNA-seq) technology that would provide the developmental changes and major biological process involved in the function of MPs after PNI.Methods:Twenty-seven male SD rats (200-300 g in weight) were selected from the Department of Hand and Foot Microscopy and Wound Repair Surgery, Henan Provincial People's Hospital (People's Hospital of Zhengzhou University) and the Department of Orthopaedics of First Affiliated Hospital of Zhengzhou University from July 2023 to December 2023. The rats were divided into a Sham operation group (Sham group), a 3 days post crush group (3 dpc group) and a 7 days post crush group (7 dpc group), following the randomised table method with 9 rats per group. After 7 days of environmental acclimatisation, the 3 dpc group and 7 dpc group were subjected to have the right sciatic nerve crushed in order to create a model of crush injury. And as a control group, the Sham group was subjected to Sham surgery only. Nine right sciatic nerves of rats were collected from each group at the corresponding time pints. Single-cell isolation was performed on the 10X Genomics platform. ScRNA-seq libraries were constructed using the Gel Bead Kit V3 and the libraries were sequenced using an Illumina Novaseq 6000 sequencer. Dimensionality reduction was performed using Principal Component Analysis and T-Distributed Stochastic Neighbor Embedding to visualise and explore the cellular heterogeneity within the dataset. Nine distinct cell clusters and their corresponding marker genes were identified based on the dimensionality-reduced data. Differential gene expression analysis was then performed to identify differentially expressed genes (DEGs) in MPs between different groups. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed to uncover the biological processes and pathways based on the DEGs. Monocle program for pseudo-time analysis was used to infer the developmental trajectory of MPs after injury.Results:A total of 19 054 cells were obtained by sequencing, and the results showed that the proportion of MPs in peripheral nerves was significantly up-regulated after PNI, and MPs were classified into 9 cellular subgroups based on the clustering analysis of the scRNA-seq data, which were Cluster 1 (3 398 cells), Cluster 2 (3 388 cells), Cluster 3 (3 262 cells), Cluster 4 (2 825 cells), Cluster 5 (2 753 cells), Cluster 6 (1 894 cells), Cluster 7 (648 cells), Cluster 8 (492 cells) and Cluster 9 (394 cells), respectively. Based on the expression of different cell subpopulation markers, MPs in the Sham group, 3 dpc group and 7 dpc group of sciatic nerves were classified into 9 cell clusters and the distributions of different MPs clusters in the 9 sciatic nerve samples were identified, among which, the Sham group had the lowest number of MPs cells in the sciatic nerve samples (a total of 2 719 cells) and the clusters were mainly dominated by clusters 5 (1 119 cells) and clusters 6 (1 240 cells). The 3 dpc group had the highest number of MPs cells (9 760 cells in total) and the clusters were mainly dominated by cluster 2 (1 760 cells), cluster 3 (3 130 cells) and cluster 4 (2 300 cells). The MPs (6 575 cells in total) in the 7 dpc group were mainly dominated by cluster 1 (2 406 cells) and cluster 2 (1 628 cells). Compared with the Sham group, the GO and KEGG annotations of the DEGs were significantly upregulated in the 3 dpc group, indicating that MPs in the rat sciatic nerves would have the ability to bind to extracellular molecules and remove debris from the injury site at 3 days post-injury, and the 7 dpc group would have the ability to activate the signalling pathways related to nerve repair. The proposed time-series analysis revealed that, in the uninjured condition, the MPs were mainly in the cluster 5 (Ccl17 +Cd80 +) and cluster 6 (Fcmr +Slc9a9 +). At 3 days post-injury, MPs developed into cell types dominated by cluster 2 (Cd8b +Meis3 +), cluster 3 (Il10 +Cd163 +) and cluster 4 (Ccl24 +Prg4 +). At 7 days post-injury, the effector state of cluster 2 among the main cell types of MPs was still maintained but the other parts had developed into cluster 1 (Hspa1b +Apobec1 +) related phenotypes. Conclusion:The molecular characteristics of MPs in the peripheral nerve revealed through scRNA-seq data provide valuable insights into the role of MPs in mediating inflammation and neural regeneration after PNI.

Ultra-rapid cooling and rewarming rate is a critical technical approach to achieve ice-free cells during the freezing and melting process. A set of ultra-rapid solid surface freeze-thaw visualization system was developed based on a sapphire flim, and experiments on droplet freeze-thaw were carried out under different cryoprotectant components, volumes and laser energies. The results showed that the cooling rate of 1 μL mixed cryoprotectant [1.5 mol/L propylene glycol (PG) + 1.5 mol/L ethylene glycol (EG) + 0.5 mol/L trehalose (TRE)] could be 9.2×10 ³ °C/min. The volume range of 1-8 μL droplets could be vitrified. After comparing the proportions of multiple cryoprotectants, the combination of equal proportion mixed permeability protectant and trehalose had the best vitrification freezing effect and more uniform crystallization characteristics. During the rewarming operation, the heating curve of glassy droplets containing gold nanoparticles was measured for the first time under the action of 400-1 200 W laser power, and the rewarming rate was up to the order of 10 ⁶ °C/min. According to the droplet images of different power rewarming processes, the laser power range for ice-free rewarming with micron-level resolution was clarified to be 1 400-1 600 W. The work of this paper simultaneously realizes the ultra-high-speed temperature ramp-up, transient visual observation and temperature measurement of droplets, providing technical means for judging the ice free droplets during the freeze-thaw process. It is conducive to promoting the development of ultra-rapid freeze-thaw technology for biological cells and tissues.
Freezing ; Vitrification ; Cryopreservation/methods* ; Trehalose ; Gold ; Rewarming ; Metal Nanoparticles ; Cryoprotective Agents ; Lasers

Objective:To explore the effect of sciatic nerve derived exosomes(SN-EXO) on axon regeneration and functional recovery after peripheral nerve injury(PNI).Methods:From March 2021 to October 2022, the Department of Orthopedics of the First Affiliated Hospital of Zhengzhou University studied the effect of SN-EXO on the proliferation of Schwann cells(SCs) through EdU cell proliferation experiment. Twenty-one healthy male SD rats were randomly divided into 3 groups of sham operation, peripheral nerve injury(PNI) and SN-EXO treatment, with 7 rats in each group. The right sciatic nerves of rat models in sham group were exposed without injury. In the rat in PNI group and SN-EXO treatment group, PBS and SN-EXO were injected under the epineurium of right sciatic nerves following sciatic nerve crush. Sciatic nerve function index(SFI) was performed at 28 days after operation, and then sacrificed. Right sciatic nerves were removed for further exploration of nerve regeneration. The histopathological changes and axon arrangement of sciatic nerves were evaluated by HE staining. Regeneration efficiency of neurofilaments and SCs were obserred by NF200 and S100β double staining of sciatic nerve. The data obtained were statistically analyzed, and P<0.05 was statistically significant. Results:It was found that SN-EXO can significantly enhance the proliferation ability of SCs, with statistically significant difference( P<0.05). SFI in SN-EXO treatment group and PNI group were(-27.65±4.36) and(-57.33±7.49), respectively, and the difference was statistically significant( P<0.05). Axons in SN-EXO treatment group were arranged more closely and orderly than those in the PNI group at 28 days after operation, and there were less injury induced axon disintegration and vacuolation. Immunofluorescence assay indicated that NF200 and S100β fluorescence intensity in SN-EXO treatment group was significantly higher than that in the PNI group, and the difference was statistically significant( P<0.05). Conclusion:SN-EXO could enhance the proliferation of SCs to promote axon regeneration following peripheral nerve injury.