Background Prenatal exposure to fine particulate matter (PM_2.5) is closely associated with cortical damage and neuroinflammation in offspring. The cyclic guanosine monophosphate–adenosine monophosphate synthase (cGAS)–stimulator of interferon genes (STING) signaling pathway is a key regulator of inflammation and may be subject to epigenetic regulation. Objective To investigate the role of cGAS-STING pathway activation in PM_2.5-induced cortical damage in offspring mice during pregnancy and the underlying epigenetic regulatory mechanisms. Methods Open field tests were used to assess depressive-like behavior in offspring mice. Morphological analysis was conducted to evaluate cortical damage and microglial activation in offspring brains. Real-time fluorescent quantitative PCR (RT-qPCR) and Western blot (WB) were performed to detect changes in the expression of key molecules in the cGAS-STING pathway in cortical tissue. A PM_2.5-induced microglial cell injury model was established in BV2 cells. Microglial activation was observed, cell viability was measured using the Cell Counting Kit-8 (CCK-8), and the expression levels of inducible nitric oxide synthase (iNOS) and key molecules in the cGAS-STING pathway were detected by RT-qPCR and WB. Bioinformatics analysis was performed to explore the epigenetic regulatory association between the STING signaling pathway and lysine-specific demethylase 5A (KDM5A). Changes in KDM5A mRNA and protein expression, as well as the protein level of histone H3 lysine 4 trimethylation (H3K4me3), were detected in an in vitro PM_2.5 injury model. Using small interfering RNA (siRNA) technology, the KDM5A gene was silenced in BV2 cells exposed to PM_2.5. The protein expression of H3K4me3 was detected to evaluate improvements in microglial activation, changes in inflammatory markers such as iNOS and mannose receptor (CD206), and alterations in the cGAS-STING pathway. Results Compared with the control group, the total distance of offspring mice in the PM_2.5 group was significantly reduced, and both the distance traveled and the time spent in the central area of the open field were significantly decreased (P<0.01, P<0.001), indicating depressive-like behavior in the offspring mice. Compared with the control group, the offspring mice in the PM_2.5 group exhibited disorganized cortical structure and significantly activated microglia (P<0.01), with significantly increased mRNA and protein levels of cGAS and STING (P<0.05, P<0.01, or P<0.001). The in vitro experiments demonstrated that the PM_2.5 treatment induced BV2 cells to polarize toward the M1 phenotype, exhibiting a distinct amoeboid morphology, with upregulated expression of the pro-inflammatory factor iNOS (P<0.05, P<0.01, or P<0.001) and activation of the cGAS-STING pathway (P<0.05, P<0.01). The analysis of RNA-seq data from KDM5A knockout cells revealed significantly downregulated STING expression, suggesting that KDM5A may activate the STING signaling pathway. The in vitro experiments further confirmed that the PM_2.5-treated BV2 cells exhibited significantly elevated mRNA and protein levels of KDM5A (P<0.01), while the H3K4me3 protein levels were markedly reduced (P<0.05). After silencing KDM5A in BV2 cells exposed to PM_2.5, compared with the PM_2.5+siNC group, the PM_2.5+siKDM5A group showed no obvious microglial activation and polarized toward the M2 phenotype, with significantly decreased expression levels of iNOS, cluster of differentiation 16 (CD16), and interleukin-1β (P<0.05, P<0.01), and significantly increased expression levels of anti-inflammatory factors CD206, YM1, and interleukin-10 (P<0.01, P<0.001). Meanwhile, the expression levels of cGAS and STING were also reduced (P<0.05, P<0.01). Conclusion KDM5A activates microglia through the cGAS-STING pathway, thereby contributing to PM_2.5-induced cortical damage in offspring mice during pregnancy.

Objective:To analyze the relationship among serum levels of fibroblast growth factor 23 (FGF23), 25-hydroxyvitamin D (25-OH-VD) and osteoporosis in children.Methods:Eighty children with osteoporosis admitted to Affiliated Hosptial of Jining Medieal University from Jun. 2019 to Jun. 2024 were included as the observation group, and 60 healthy children who underwent physical examination during the same period were included as the control group. Serum FGF23 and 25-OH-VD levels were compared between the two groups, and bone mineral density and bone metabolism indexes [CTX-1, osteocalcin (OC) and N-terminal propeptide of type I precollagen (PINP) ] were detected. Pearson correlation was used to analyze the correlation among serum FGF23, 25-OH-VD, bone metabolism indexes and bone mineral density. Logistic regression model was used to analyze the risk factors of osteoporosis in children.Results:Serum FGF23 and CTX-1 levels in the observation group were significantly higher than those in the control group ( t=15.77, 7.56, P < 0.05), while serum 25-OH-VD, OC, PINP and BMD levels were significantly lower ( t=14.09, 2.70, 13.25, 3.63, P < 0.05) ; Pearson correlation analysis showed that BMD was not correlated with age, BMI and OC ( r=-0.07, -0.02, 0.01, P > 0.05), BMD was negatively correlated with FGF23 and CTX-1, and positively correlated with 25-OH-VD and PINP ( r=-0.35, -0.34, 0.41, 0.40, P < 0.05). The stepwise regression model showed that FGF23 and 25-OH-VD were the main factors affecting BMD ( t=-2.40, 9.02, P<0.05). Multifactor Logistic regression model showed that FGF23 and 25-OH-VD were related factors of osteoporosis in children ( OR=3.01,1.16, P<0.05) . Conclusion:Serum FGF23 level is higher and 25-OH-VD level is lower in children with osteoporosis, which is significantly correlated with bone mineral density and is a related factor for osteoporosis in children.

Objective To investigate the function of Ring finger protein 13(RNF13)in cerebral ischemia reperfusion injury(CIRI),and its mechanism.Methods The mouse middle cerebral artery embolization and primary neuron oxygen-glucose depri-vation and reoxygenation were used as disease models.The CRISPR/Cas9 gene knockout technique,immunohistochemical stai-ning,immunofluorescence staining,Western blot and lipid peroxidation detection were used to evaluate the regulatory effect and molecular mechanism of RNF13 on ferroptosis in CIRI.Student's t test was used for the comparison of two samples,and one-way analysis of variance was used for the comparison of multiple samples.Results The expression levels of RNF13 protein in mice and primary neurons were upregulated during CIRI.After knockout of RNF13,ferritin heavy chain 1(Fth1)and ferritin light chain(Ftl)were downregulated,and the content of free ferrous ions and the accumulation of lipid peroxides in mice brain tissues were promoted,leading to ferroptosis aggravation and neurological impairments.Overexpression of RNF13 protected a-gainst ferroptosis by reducing the production of free ferrous and lipid peroxides in neurons.Conclusion RNF13 alleviates fer-roptosis in neurons after CIRI,and the effect is induced by Fth1.

Objective:To analyze the relationship among serum levels of fibroblast growth factor 23 (FGF23), 25-hydroxyvitamin D (25-OH-VD) and osteoporosis in children.Methods:Eighty children with osteoporosis admitted to Affiliated Hosptial of Jining Medieal University from Jun. 2019 to Jun. 2024 were included as the observation group, and 60 healthy children who underwent physical examination during the same period were included as the control group. Serum FGF23 and 25-OH-VD levels were compared between the two groups, and bone mineral density and bone metabolism indexes [CTX-1, osteocalcin (OC) and N-terminal propeptide of type I precollagen (PINP) ] were detected. Pearson correlation was used to analyze the correlation among serum FGF23, 25-OH-VD, bone metabolism indexes and bone mineral density. Logistic regression model was used to analyze the risk factors of osteoporosis in children.Results:Serum FGF23 and CTX-1 levels in the observation group were significantly higher than those in the control group ( t=15.77, 7.56, P < 0.05), while serum 25-OH-VD, OC, PINP and BMD levels were significantly lower ( t=14.09, 2.70, 13.25, 3.63, P < 0.05) ; Pearson correlation analysis showed that BMD was not correlated with age, BMI and OC ( r=-0.07, -0.02, 0.01, P > 0.05), BMD was negatively correlated with FGF23 and CTX-1, and positively correlated with 25-OH-VD and PINP ( r=-0.35, -0.34, 0.41, 0.40, P < 0.05). The stepwise regression model showed that FGF23 and 25-OH-VD were the main factors affecting BMD ( t=-2.40, 9.02, P<0.05). Multifactor Logistic regression model showed that FGF23 and 25-OH-VD were related factors of osteoporosis in children ( OR=3.01,1.16, P<0.05) . Conclusion:Serum FGF23 level is higher and 25-OH-VD level is lower in children with osteoporosis, which is significantly correlated with bone mineral density and is a related factor for osteoporosis in children.

Objective To investigate the function of Ring finger protein 13(RNF13)in cerebral ischemia reperfusion injury(CIRI),and its mechanism.Methods The mouse middle cerebral artery embolization and primary neuron oxygen-glucose depri-vation and reoxygenation were used as disease models.The CRISPR/Cas9 gene knockout technique,immunohistochemical stai-ning,immunofluorescence staining,Western blot and lipid peroxidation detection were used to evaluate the regulatory effect and molecular mechanism of RNF13 on ferroptosis in CIRI.Student's t test was used for the comparison of two samples,and one-way analysis of variance was used for the comparison of multiple samples.Results The expression levels of RNF13 protein in mice and primary neurons were upregulated during CIRI.After knockout of RNF13,ferritin heavy chain 1(Fth1)and ferritin light chain(Ftl)were downregulated,and the content of free ferrous ions and the accumulation of lipid peroxides in mice brain tissues were promoted,leading to ferroptosis aggravation and neurological impairments.Overexpression of RNF13 protected a-gainst ferroptosis by reducing the production of free ferrous and lipid peroxides in neurons.Conclusion RNF13 alleviates fer-roptosis in neurons after CIRI,and the effect is induced by Fth1.

Objective To explore the effect of ropivacaine combined with dexmedetomidine in transversus abdominis plane block(TAPB)on postoperative stress hormones and cognitive function in patients undergoing laparoscopic radical gastrectomy.Methods A total of 80 patients undergoing laparoscopic radical gastrectomy at the First Affiliated Hospital of Xinxiang Medical University from April to October 2023 were selected as the research subjects.According to different anesthesia methods,the patients were divided into the observation group and the control group,with 40 patients in each group.Patients in the observation group were injected bilaterally with 2.5 g·L-1 ropivacaine and 0.5 μg·kg-1 dexmedetomidine for TAPB,with 20 mL injection on each side.Patients in the control group were injected bilaterally with 2.5 g·L-1 ropivacaine for TAPB,with 20 mL injection on each side.Mean arterial pressure(MAP)and heart rate(HR)were recorded at the time of admission to the operating room(T1),immediately after endotracheal intubation(T2),40 minutes after pneumoperitoneum(T3),and 15 minutes after extubation(T4).Radioimmunoprecipitation was used to detect serum cortisol(COR)level,and enzyme-linked immunosorbent assay was used to measure serum norepinephrine(NE)and epinephrine(E)levels at 1,6,12,and 24 hours after surgery.Visual analog scale(VAS)was used to assess pain at rest,and Ramsay sedation scale(RSS)was used to evaluate sedation depth.The doses of propofol and sufentanil were compared between the two groups.Serum β-amyloid(Aβ)and S100β protein levels at 1 day before surgery,1 and 3 days after surgery were detected by using the enzyme-linked immunosorbent assay,and cognitive function was assessed at the same time points by using the mini-mental state examination(MMSE).Results At T,and T2,there was no significant difference in MAP and HR between the control group and the observation group(P＞0.05).At T3 and T4,MAP and HR in the observation group were significantly lower than those in the control group(P＜0.05).At 1,6,and 12 hours postoperatively,VAS score in the observation group was significantly lower than that in the control group(P＜0.05).At 24 hours postoperatively,there was no significant difference in VAS score between the control group and observation group(P＞0.05).At 1 and 6 hours postoperatively,RSS score in the observation group was significantly higher than that in the control group(P＜0.05).At 12 and 24 hours postoperatively,there was no significant difference in RSS score between the control group and observation group(P＞0.05).At 1,6,and 12 hours postoperatively,COR,NE,and E levels in the observation group were significantly lower than those in the control group(P＜0.05).At 24 hours postoperatively,there was no significant difference in COR,NE,and E levels between the control group and observation group(P＞0.05).The doses of propofol and sufentanil in the observation group were significantly lower than those in the control group(P＜0.05).One day before surgery,there was no significant difference in MMSE score between the control group and observation group(P＞0.05).At 1 and 3 days postoperatively,MMSE score in the observation group was significantly higher than that in the control group(P＜0.05).One day before surgery,there was no significant difference in serum Aβ and S100β protein levels between the control group and observation group(P＞0.05).At 1 and 3 days postoperatively,serum Aβ and S100β protein levels in the observation group were significantly lower than those in the control group(P＜0.05).Conclusion Ropivacaine combined with dexmedetomidine in TAPB in radical gastrectomy can significantly reduce postoperative pain,increase sedative effect,prolong the duration of TAPB,and benefit patients'postoperative recovery with reduced cognitive impairment.

Objective:To investigate the effects of long-term exposure to chrysotile and crocidolite on miRNAs and epithelial mesenchymal transformation (EMT) -related gene expression in human pleural mesothelial cells.Methods:In November 2020, fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of EMT-related genes in human pleural mesothelioma cells (NCl-H2052 cells, NCl-H2452 cells) and human normal mesothelial cells (Met-5A cells). MiRNAs with abnormal expression in human pleural mesothelioma cells were screened out from the previous miRNA chip data of research group, and target genes of differentially expressed miRNAs were predicted using miRWalk database (http: //mirwalk.umm.uni-heidelberg.de). RT-qPCR was used to verify the abnormal expression of EMT-related miRNAs in cell lines. Met-5A cells were treated with 5μg/cm 2 chrysotile and crocidolite respectively for 48 h a time, once a week and a total of 10 times. Chrysotile group, crocidolite group and control group were set up. And the control group was added with the same volume of PBS. The expression changes of EMT-related genes and abnormal expression miRNAs in each group were detected by RT-qPCR. The differences among the groups were compared by one-way ANOVA, and the differences between the control group and the experimental group were compared by dunnet- t test. Results:Compared with Met-5A cells, the expression levels of Vimentin and Twist genes were increased, and the expression level of E-cadherin genes was decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001). Target genes of miRNAs with abnormal expression in miRNA chip were predicted, and the results showed four abnormally expressed miRNAs associated with EMT and verified the expression of these four miRNAs in the cell lines. Compared with Met-5A cells, the expression level of hsa-miR-155-5p was increased in NCl-H2052 cells and NCl-H2452 cells, the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001), which was consistent with the results of chip analysis. After exposure of Met-5A cells, it was found that compared with the control group, the expression levels of Vimentin and Twist genes, hsa-miR-155-5p, hsa-miR-34b-5p and hsa-miR-34c-5p in the crocidolite group were increased, while the expression level of E-cadherin gene was decreased ( P<0.05). Compared with the control group, the expression levels of Vimentin, Twist and E-cadherin genes in chrysotile group were increased, while the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased ( P<0.05) . Conclusion:Long-term exposure to chrysotile and crocidolite could cause Met-5A cells to produce miRNAs and EMT-related gene expression changes similar to mesothelioma cells.

Objective:To demonstrate the three-dimensional structures of the limbic system and its fiber connections through fiber dissection,and to provide reference for relevant professionals to master the anatomy of the limbic system.Methods:Ten cerebral hemispheres were treated and dissected according to Kelinger method,and the limbic system and its fiber connections were displayed.Results:The limbic system was arranged around the thalamus and corpus cal-losum in a double-layer concentric circle structure.The outer layer structures mainly consisted of the cingulate gyrus and the parahippocampal gyrus,while the inner layer structures included the amygdala,hippocampus and fornix.The main association fiber of the outer layer is the cingulum,whose superior trunk is mainly located in the cingulate gyrus,and the inferior trunk is mainly located in the parahippocampal gyrus.The fiber structures of the inner layer includes the striae terminalis and ansa peduncularis emanating from the amygdala and the fornix of the hippocampus.Conclusion:Limbic system is an important connection structure between telencephalon and diencephalon,and its anatomical struc-ture is complex.Fiber dissection method can effectively demonstrate the complex spatial structure of limbic system,which is of great benefit to relevant professionals to understand its three-dimensional structure.

Objective: To investigate the screening results of adolescent scoliosis in Chuzhou City, Anhui Province, and analyze the influencing factors for scoliosis, so as to provide insights into the prevention and control of scoliosis among adolescents. Methods: Students were selected from six primary and middle schools in Chuzhou City using the stratified random cluster sampling method from April to June 2023. Demographic information, daily behaviors and postures, and exercise status were collected through questionnaire surveys. Scoliosis was screened and diagnosed according to the Screening for Abnormal Spinal Curvature in Children and Adolescents. Influencing factors for scoliosis among primary and middle school students were identified using a multivariable logistic regression model. Results: A total of 1 823 questionnaires were allocated, and 1 768 effective questionnaires were recovered, with an effective response rate of 96.98%. There were 537 primary school students, 1 000 junior high school students and 231 senior high school students, with an average age of (13.40±1.92) years. There were 948 male students (53.62%) and 820 female students (46.38%). A total of 131 cases of scoliosis were screened positive, with a positive rate of 7.41%. The results of multivariable logistic regression analysis showed that gender (female, OR=1.759, 95%CI: 1.135-2.727), body mass index (OR=0.593, 95%CI: 0.538-0.654), sleeping position (side lying, OR=0.598, 95%CI: 0.377-0.951; prone lying, OR=2.336, 95%CI: 1.201-4.545), frequency of reading in bed (often, OR=2.099, 95%CI: 1.201-3.670), way of carrying backpack (shoulders, OR=0.580, 95%CI: 0.370-0.908), and exercise level (OR=0.428, 95%CI: 0.296-0.618) were influencing factors of scoliosis among primary and middle school students. Conclusion The positive rate of scoliosis screening among primary and middle school students in Chuzhou City was 7.41%, which was influenced by gender, age, body mass index, sleeping posture, reading in bed, way of carrying backpack and exercise level.

Objective To investigate the role of macrophages in the process of fine particulate matter (PM2.5)exposure induced damage to pulmonary blood-air barrier.Methods Eighteen male BALB/C mice (aged of 10 weeks,weighing 24～27 g)were randomly divided into control group and low-and high-dose PM2.5 exposure groups (receiving 1 .8 and 16.2 mg/kg,respectively),with 6 mice in each group.The control group received tracheal instillations of normal saline on days 1,4,and 7,whereas the exposure groups were administered corresponding dose of PM2.5 exposure at the same time points.In 24 h after last exposure,pathological changes in the lung tissues were observed,and the contents of total protein (TP ),lactate dehydrogenase (LDH ),and alkaline phosphatase (AKP ) in bronchoalveolar lavage fluid (BALF ),and F4/80 protein level in lung tissue were measured to evaluate the blood-air barrier damage and macrophage infiltration within the lung tissues.Additionally,an in vitro model of the blood-air barrier was established using A549 alveolar epithelial cells and EA.hy926 vascular endothelial cells.In combination with a THP-1 macrophage model,the supernatant PM2.5 supernatant,macrophage supernatant,and PM2.5-macrophage supernatant were incubated with the barrier model for 24 h,respectively.Transmembrane electrical resistance (TEER),sodium fluorescein permeability of the barrier model,and LDH release from the barrier cells were measured to ascertain the extent of macrophage-mediated enhancement in barrier damage induced by PM2.5 exposure.Furthermore,the expression of inflammatory cytokines,such as TNF-α,IL-1β,IL-6,and IL-8 in the macrophages after PM2.5 exposure was analyzed with quantitative real-time PCR (qPCR)and enzyme-linked immunosorbent assay (ELISA).Results PM2.5 exposure induced lung tissue damage in mice in a dose-dependent manner,significantly elevated the contents of TP,LDH and AKP in the BALF and caused marked infiltration of macrophages into the lung tissue,especially the high-dose exposure when compared with the mice from the control group (P<0.01 ).In vitro barrier model exposure experiments showed that in comparison with the treatment of 150 and 300 μg/mL PM2.5 and macrophage supernatant,the same doses of PM2.5-macrophage supernatant resulted in notably decreased TEER and significantly enhanced permeability in the barrier model (P<0.01 ),and markedly increased LDH release from epithelial and endothelial barrier cells (P<0.01 ).Additionally,the exposure of 150 and 300μg/mL PM2.5 led to a significant up-regulation of TNF-α,IL-1β,IL-6,and IL-8 in the macrophages (P<0.01 ).Conclusion Macrophages deteriorate PM2.5-induced functional impairment of the pulmonary blood-air barrier.