OBJECTIVES: Neuropathic pain (NP) is one of the most common forms of chronic pain, yet current treatment options are limited in effectiveness. Peripheral nerve injury activates spinal microglia, altering their inflammatory response and phagocytic functions, which contributes to the progression of NP. Most current research on NP focuses on microglial inflammation, with relatively little attention to their phagocytic function. Early growth response factor 2 (EGR2) has been shown to regulate microglial phagocytosis, but its specific role in NP remains unclear. This study aims to investigate how EGR2 modulates microglial phagocytosis and its involvement in NP, with the goal of identifying potential therapeutic targets. METHODS: Adult male Sprague-Dawley (SD) rats were used to establish a chronic constriction injury (CCI) model of the sciatic nerve. Pain behaviors were assessed on days 1, 3, 7, 10, and 14 post-surgery to confirm successful model induction. The temporal and spatial expression of EGR2 in the spinal cord was examined using real-time quantitative PCR (RT-qPCR), Western blotting, and immunofluorescence staining. Adeno-associated virus (AAV) was used to overexpress EGR2 in the spinal cord, and behavioral assessments were performed to evaluate the effects of EGR2 modulation of NP. CCI and lipopolysaccharide (LPS) models were established in animals and microglial cell lines, respectively, and changes in phagocytic activity were measured using RT-qPCR and fluorescent latex bead uptake assays. After confirming the involvement of microglial phagocytosis in NP, AAV was used to overexpress EGR2 in both in vivo and in vitro models, and phagocytic activity was further evaluated. Finally, eukaryotic transcriptome sequencing was conducted to screen differentially expressed mRNAs, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses to identify potential downstream effectors of EGR2. RESULTS: The CCI model successfully induced NP. Following CCI, EGR2 expression in the spinal cord was upregulated in parallel with NP development. Overexpression of EGR2 via spinal AAV injection enhanced microglial phagocytic activity and increased pain hypersensitivity in rats. Both animal and cellular models showed that CCI or LPS stimulation enhanced microglial phagocytosis, which was further amplified by EGR2 overexpression. Transcriptomic analysis of spinal cord tissues from CCI rats overexpressing EGR2 revealed upregulation of numerous genes associated with microglial phagocytosis and pain regulation. Among them, Lag3 emerged as a potential downstream target of EGR2. CONCLUSIONS EGR2 contributes to the maintenance of NP by enhancing microglial phagocytosis in the spinal dorsal horn.
Animals ; Microglia/metabolism* ; Phagocytosis/physiology* ; Rats, Sprague-Dawley ; Neuralgia/physiopathology* ; Early Growth Response Protein 2/metabolism* ; Male ; Rats ; Spinal Cord/metabolism* ; Sciatic Nerve/injuries*

BackgroundPrevious studies on subgroups of adolescent non-suicidal self-injury （NSSI） behaviors have predominantly focused on community and school-based samples， with limited research on clinical samples. The precise identification of clinically distinct NSSI subgroups in adolescents is critical for optimizing medical resource allocation. ObjectiveTo explore the subgroups of NSSI in adolescent outpatients， and to provide references for intervening of NSSI behaviors. MethodsFrom May 2021 to April 2022， 192 adolescents who met the NSSI diagnostic criteria as outlined in the Diagnostic and Statistical Manual of Mental Disorders， fifth edition （DSM-5） were recruited from psychiatry or psychology clinics at 6 hospitals in Fujian Province. Participants were assessed using the Adolescent Non-suicidal Self-injury Assessment Questionnaire （ANSAQ）， the Childhood Trauma Questionnaire （CTQ）， the Depression Anxiety Stress Scale-21 item（DASS-21） and the Difficulties in Emotion Regulation Scale （DERS）. Latent profile analysis was conducted using Mplus 8.0. ResultsA total of 192 adolescents with NSSI behaviors completed the study， and the analysis yielded three subgroups of self-injurers： the mild self-injury group （50 cases， 26.04%）， the emotional disorder group （81 cases， 42.19%）， and the severe self-injury group （61 cases， 31.77%）. There were significant differences in the total scores of CTQ， DASS-21 and DERS among the three groups （F=8.058， 51.414， 48.742， P<0.01）. The severe self-injury group exhibited significantly higher scores in CTQ， DASS-21 and DERS when compared to both the mild self-injury group and the emotional disorder group （P<0.05）. Moreover， the emotional disorder group also reported notably higher DASS-21 and DERS scores in comparison to the mild self-injury group （P<0.05）. ConclusionClinical studies on adolescent outpatients engaging in NSSI behaviors reveal three distinct typological profiles， mild self-injury， emotional disorder-related self-injury， and severe self-injury. ［Funded by Fujian Social Science Planning Project （number， FJ2019B173）］

Objective:To explore the mechanism by which miRNA-4469 (miR-4469) regulates the proliferation and invasion of renal cancer cells in vitro.Methods:The survival differences of patients with different expression levels of miR-4469 were analyzed based on the OncomiR database. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) method was used to detect the expression of miR-4469 in renal cancer cell lines ACHN, OS-RC-2, SK-RC-20, 769-P, A498 and normal renal tubular epithelial cell line HK-2, and the renal cancer cells with the lowest expression level of miR-4469 were divided into miR-4469 group and control group, and were transfected with miR-4469 mimic and negative control sequence, respectively. The CCK-8 assay was used to detect the cell proliferation ability (expressed as absorbance value) in the two groups, and Transwell assay was used to analyze the number of invasive cells in the two groups. TargetScan Release 8.0 software was used to predict the binding site between miR-4469 and protein disulfide isomerase A4 (PDIA4) mRNA, and dual-luciferase reporter gene assay was used to verify the targeting relationship between miR-4469 and PDIA4 mRNA. qRT-PCR method was used to detect the expression of PDIA4 mRNA in cells of each group, and Western blotting method was used to detect the expression levels of PDIA4 protein and PI3K-AKT-m-TOR pathway proteins in cells of each group.Results:Analysis of relevant data from the OncomiR database showed that compared with patients with low miR-4469 expression, the overall survival of renal cancer patients with high miR-4469 expression was better ( P < 0.001). The relative expression of miR-4469 in each renal cancer cell line was lower than that in HK-2 cells (all P < 0.05), and the expression of miR-4469 in 769-P cells was the lowest, which were selected to perform the subsequent experiments. The proliferation ability of 769-P cells in the miR-4469 group was lower than that in the control group ( P < 0.01). The number of 769-P cell invasions in the miR-4469 group were less than that in the control group [(19±3) cells vs. (64±7) cells, t = 5.44, P = 0.002]. Compared with the co-transfection of wild-type PDIA4 and miR-4469 negative sequence group, the relative luciferase activity of cells in the co-transfection of wild-type PDIA4 and miR-4469 mimic sequence group was lower (0.42±0.07 vs. 1.01±0.08, t = 5.74, P = 0.001); there was no statistical difference in cell luciferase activity between the co-transfected mutant PDIA4 and miR-4469 negative sequence group and the co-transfected mutant PDIA4 and miR-4469 mimic sequence group (0.99±0.11 vs. 1.02±0.11, t = 0.19, P = 0.001). The relative expression levels of PDIA4 mRNA in 769-P cells in the miR-4469 group were lower than that in the control group (0.98±0.23 vs. 7.19±2.23, t = 2.77, P = 0.032). Compared with the control group, the expression of PDIA4 protein and PI3K-AKT-m-TOR pathway-related p-PI3K, p-AKT, p-mTOR, and p-SGK1 proteins in 769-P cells in the miR-4469 group were all lower (all P < 0.05). Conclusions:miR-4469 may be related to the survival of renal cancer patients, and its expression is down-regulated in various renal cancer cell lines. miR-4469 may inhibit the proliferation and invasion of renal cancer 769-P cells by regulating the PI3K-AKT-m-TOR pathway through PDIA4.

Objective:To explore the mechanism of Ma-Xing Shi-Gan decoction combined with Xiao-Chai-Hu decoction (hereinafter referred to as " Sanyang combined treatment" ) in alleviating colon injury in mice infected with influenza virus by transcriptome sequencing technique.Methods:The mouse model of colonic injury caused by influenza virus was induced by intranasal drip of influenza A virus H1N1 suspension. The mice were divided into Control group, Model group, and Sanyang combined treatment (SCT) group. Model group and SCT group were fed with PBS and Ma-Xing Shi-Gan decoction combined with Xiao-Chai-Hu decoction respectively. Seven days later, the colon tissues of each group were taken, the colon length and pathological damage were observed, and the transcriptome was sequenced to screen the significantly different genes between the SCT group and model group for Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Set Enrichment Analysis(GSEA).Results:After the therapy with SCT, the length of the colon of mice was significantly improved and the pathological injury of the colon was reduced. There are 92 differentially expressed genes between the SCT group and the model group. GO analysis indicated that the differential genes were enriched in biological processes such as regulation of cytokine and chemokine production, inflammatory response, defense response, immune response, regulation of NF-κB inducing kinase(NIK)/Nuclear factor-κB(NF-κB) signal and Mitogen-activated protein kinase(MAPK) cascade, as well as cell components related to intestinal barrier such as brush border membrane, brush border and microvilli. KEGG analysis indicated that the differential genes were enriched in Toll-like receptor signaling pathway, intestinal immune network for IgA production, complement and coagulation cascade, and Peroxisome proliferator-activated receptor(PPAR) signaling pathway. GSEA indicated that the intestinal immune network for IgA production, PPAR signaling pathway, propionic acid metabolism and butyrate metabolism were significantly up-regulated after the intervention with SCT, while apoptosis and MAPK signaling pathway were significantly down-regulated.Conclusions:Sanyang combined therapy can protect the intestinal tract of mice infected with influenza virus mainly through immunity, inflammation and metabolism pathways.

Humans ; Adult ; Serum Albumin, Human ; Consensus ; Cardiac Surgical Procedures ; Thoracic Surgery

Objective:To explore the relationship between the relative expression of miRNA-676-3p and the survival of renal cancer patients, and its effect on the proliferation and invasion of renal cancer by targeting and regulating prefoldin 1 (PFDN1).Methods:OncoRank online software was selected to analyze the relationship between the relative expression of miRNA-676-3p and the survival rate of renal cancer patients. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the relative expression of miRNA-676-3p in renal cancer cell lines. Renal carcinoma CAKI1 cells were resuscitated, and the transfected miRNA-NC was used as the control group, and the transfected precursor miRNA-676-3p was used as the overexpression group. The relative expression of miRNA-676-3p was detected by RT-qPCR. The cell absorbance and invasion number of the two groups were measured by CCK-8 and Transwell invasion assays, respectively. The target gene of miRNA-676-3p was predicted and verified by referring to the TargetScan Release 8.0 website and dual-luciferase reporter gene experiment. The expression of PFDN1 gene and Wnt/β-catenin molecular pathway protein in the two groups of cells were determined by RT-qPCR and Western blotting, respectively. Measurement data were expressed as mean ± standard deviation ( ± s), t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:The survival rate of renal cancer patients with high expression of miRNA-676-3p was significantly higher than that of renal cancer patients with low expression of miRNA-676-3p, the difference was statistically significant ( P<0.01). The relative expression of miRNA-676-3p in renal cancer cell lines was significantly lower than that in normal renal tubular epithelial cells, the difference was statistically significant ( P<0.01), and the relative expression of miRNA-676-3p in CAKI1 cells was the lowest, the difference was statistically significant ( P<0.01). The relative expression levels of miRNA-676-3p in the control and overexpression groups were 1.04±0.59 and 15.90±1.70, respectively, and the overexpression group was significantly higher than the control group, the difference was statistically significant ( P<0.01). After 24, 48, 60, and 72 h of culture, the absorbance of cells in the overexpression group was lower than that in the control group, the difference was statistically significant ( P<0.05). The number of invasion cells in the control group and the overexpression group were (115.90 ± 24.73) and (43.83 ± 21.94) cells, respectively, and the number of cell invasion in the overexpression group was significantly lower than that in the control group, the difference was statistically significant ( P<0.01). PFDN1 was the downstream target gene of miRNA-676-3p ( P<0.01). The relative expression of PFDN1 gene in the overexpression group was significantly lower than that in the control group, the difference was statistically significant ( P<0.01). The expression of Wnt/β-catenin molecular pathway proteins in the overexpression group was lower than that in the control group. Conclusions:Renal cancer patients with high expression of miRNA-676-3p had a higher survival rate. miRNA-676-3p inhibited the proliferation and invasion of renal cancer CAKI1 cells by significantly down-regulating the expression of PFDN1, thereby inhibiting the development of renal cancer.

Objective:The relative expression of lncRNA NPIPA9 in prostate cancer tissues was analyzed, and the relative expression of miR-210-3p and its effect on the growth and migration of prostate cancer cells were detected by overexpressing lncRNA NPIPA9.Methods:The relative expression of lncRNA NPIPA9 in prostate cancer tissues was analyzed by Oncomine database. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the relative expression of lncRNA NPIPA9 in prostate cancer cell lines DU-145, PC-3, C4-2B, 22Rv1, LNCaP and normal prostate epithelial cell RWPE-1. Prostate cancer PC-3 cells were cultured in vitro and divided into control group (transfected with control vector 100 nmol/L) and NPIPA9 group (transfected with lncRNA NPIPA9 vector 100 nmol/L). The proliferation activity of PC-3 cells was detected by CCK-8 method. The migration ability of PC-3 cells was detected by Transwell method. Potential target of lncRNA NPIPA9 were predicted using bioinformatics techniques. The dual-luciferase reporter gene assay determined the target binding relationship between lncRNA NPIPA9 and miR-210-3p. The effect of lncRNA NPIPA9 on the relative expression of miR-210-3p in prostate cancer cells was detected by RT-qPCR. The effect of lncRNA NPIPA9 on the expression of nuclear factor kappa-B (NF-κB) pathway proteins in prostate cancer cells was detected by Western blotting. Measurement data were expressed as mean±standard deviation ( ± s), and t-test was used for comparison between two groups, one-way analysis of variance was used for comparison between multiple groups. Results:The expression of lncRNA NPIPA9 in prostate cancer tissue was lower than that in adjacent tissue, the difference was statistically significant ( P<0.01). The relative expression of lncRNA NPIPA9 in prostate cancer cell lines was lower than that in RWPE-1 cells, the difference was statistically significant ( P<0.01), and the relative expression of lncRNA NPIPA9 in prostate cancer PC-3 cells was the lowest, the difference was statistically significant ( P<0.01). Compared with the control group, lncRNA NPIPA9 had an inhibitory effect on the viability of prostate cancer PC-3 cells, the difference was statistically significant ( P<0.05). The migration numbers of PC-3 cells in the control group and NPIPA9 group were 101.70±8.63 and 45.97±8.83, respectively, and lncRNA NPIPA9 had an inhibitory effect on PC-3 cell migration, the difference was statistically significant ( P<0.01). lncRNA NPIPA9 can directly target miR-210-3p, the difference was statistically significant ( P<0.01). The relative expression of miR-210-3p in PC-3 cells in control group and NPIPA9 group were 5.32 ± 0.79 and 1.11 ± 0.56, respectively, and lncRNA NPIPA9 could directly down-regulate the expression of miR-210-3p in PC-3 cells, the difference was statistically significant ( P<0.01). Compared with the control group, lncRNA NPIPA9 can reduce the expression of NF-κB pathway proteins c-Myc, MMP-9, VEGF, p65, p50 in PC-3 cells. Conclusion:The expression of lncRNA NPIPA9 is down-regulated in prostate cancer tissues, and it reduces the proliferation and migration ability of prostate cancer PC-3 cells by targeting and negatively regulating miR-210-3p.

Objective:To explore the clinical and genetic characteristics of two children with a clinical diagnosis of Treacher Collins syndrome (TCS).Methods:Whole-exome sequencing was used to screen potential variants in the two children. Confirmation of suspected variants was performed through Sanger sequencing , multiplex ligation dependent probe amplification and real-time PCR in probands and their parents.Results:A heterozygous deletion variant, c. 4357_4360delGAAA, was detected in case one, while was de novo and verified by Sanger sequencing. Thevariant was classified as pathogenic(PVS1 + PM2+ PM6)according to ACMG guideline. The heterozygous deletion of exon 1-7 was seen in the same gene in case 2, which MLPA verified as heterozygous deletion of exon 1-6. This deletion was inherited from the father with a normal phenotype, and the father’s TCOF1 gene was suspected to be chimeric heterozygous deletion of exon 1-6 verified by MLPA. Conclusion:The identified variants in the TCOF1 gene probably underlie the two cases of TCS. There was no apparent correlation between genotype and phenotype. In addition, it shows a high interfamilial variability ranging from normal to full presentation of TCS. Genetic detection provided clinical diagnosis and genetic counselling for TCS patients .

Objective:To investigate the therapeutic mechanism of high-frequency repetitive transcranial magnetic stimulation (rTMS) in treating mild amnestic cognitive impairment (aMCI).Methods:Twenty-five patients with aMCI were randomly divided into an observation group of 13 and a control group of 12. The observation group was given 10-Hz rTMS over the left dorsolateral prefrontal cortex at 80% of the motor threshold-400 pulses a day, 5 times a week for 4 consecutive weeks. The control group received sham stimulation on the same schedule. Before and after the experiment, both groups were evaluated using the Montreal cognitive assessment (MoCA) and received fMRI scans.Results:After the intervention, the average MoCA score of the observation group had improved significantly more compared with that of the control group and compared with before the intervention. According to the fMRI results, regional homogeneity in the right middle frontal gyrus of the observation group had increased significantly, while that of the control group both there and in the left precuneus had decreased significantly.Conclusions:High-frequency rTMS can effectively improve the cognitive function of patients with aMCI and synchronize neuron activity in cognition-related brain regions.

Objective To explore the therapeutic effect of mild hypothermia on the inflammatory response, organ function and outcome in perioperative patients with acute Stanford type A aortic dissection (AAAD). Methods From February 2017 to February 2018, 56 patients with AAAD admitted in our department were enrolled and randomly allocated into two groups including a control group and an experimental group. After deep hypothermia circulatory arrest during operation, in the control group (n=28), the patients were rewarmed to normal body temperatures (36 to 37 centigrade degree), and which would be maintained for 24 hours after operation. While in the experimental group (n=28), the patients were rewarmed to mild hypothermia (34 to 35 centigrade degree), and the rest steps were the same to the control group. The thoracic drainage volume and the incidence of shivering at the first 24 hours after operation, inflammatory indicators and organ function during perioperation, and outcomes were compared between the two groups. There were 20 males and 8 females at age of 51.5±8.7 years in the control group, 24 males and 4 females at age of 53.3±11.2 years in the experimental group. Results There was no obvious difference in the basic information and operation information in patients between the two groups. Compared to the control group, at the 24th hour after operation, the level of peripheral blood matrix metalloproteinases (MMPs) was lower than that in the experimental group (P=0.008). In the experimental group, after operation, the awakening time was much shorter (P=0.008), the incidence of bloodstream infection was much lower (P=0.019). While the incidence of delirium, acute kidney injury (AKI), hepatic insufficiency, mechanical ventilation duration, intensive care unit (ICU) stays, or hospital mortality rate showed no statistical difference. And at the first 24 hours after operation, there was no difference in the thoracic drainage volume between the two groups, and no patient suffered from shivering. Conclusion The mild hypothermia therapy is able to shorten the awakening time and reduce the incidence of bloodstream infection after operation in the patients with AAAD, and does not cause the increase of thoracic drainage volume or shivering.