Objective:Glioblastoma (GBM) is a primary malignant tumor in the central nervous system, whose tissue heterogeneity and invasive growth characteristics lead to a very poor prognosis for patients. Hub genes in GBM are screened by bioinformatics analysis; expressions of Hub genes in human GBM tissue, and their effects on GBM biological behaviors and Notch signaling pathway proteins are explored, and their regulatory roles in GBM proliferation in xenograft tumor model mice are evaluated.Methods:(1) The gene expression profiles of GBM tissue and normal brain tissue from the Gene Expression Omnibus (GEO) database were obtained; differentially expressed genes (DEGs) were screened, and gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses were conducted on DEGs. A co-expression network was constructed using weighted gene co-expression network analysis (WGCNA) to identify modules significantly related to GBM. Gene set enrichment analysis (GSEA) on the related module genes was performed; STRING database (version 12.0) was used to construct protein-protein interaction (PPI) network and screen the hub genes. (2) Normal brain tissue samples from 8 patients with epilepsy and GBM tissue samples from 10 patients with GBM who underwent surgical resection in Department of Neurosurgery, He'nan Provincial People's Hospital from January 1, 2022 to December 1, 2023 were collected; expressions of synaptosomal-associated protein 25 (SNAP25) and vesicle-associated membrane protein 2 (VAMP2) were detected by immunohistochemical staining and Western blotting. (3) LN229 and U87 cells were routinely cultured in vitro and divided into shRNA-SNAP25/shRNA-VAMP2 group and empty vector group, and then, cells in each group were transfected with shRNA-SNAP25/shRNA-VAMP2 lentivirus or empty vector lentivirus, respectively; 24 hours after transfection, SNAP25 and VAMP2 mRNA and protein expressions in the 2 groups were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and Western blotting. LN229 and U87 cells were routinely cultured in vitro and divided into shRNA-SNAP25 group, shRNA-VAMP2 group and empty vector group; cells in each group were transfected with shRNA-SNAP25 lentivirus, shRNA-VAMP2 lentivirus or empty vector lentivirus, respectively; 48 hours after transfection, proliferation was detected by clone formation assay, proliferating cell nuclear antigen Ki-67 expression was detected by immunofluorescent staining, invasion was detected by Transwell invasion assay, and Notch1, HEY1 and HES1 protein expressions in Notch signaling pathway of LN229 cells were detected by Western blotting. (4) LN229 cells transfected with shRNA-SNAP25 lentivirus, shRNA-VAMP2 lentivirus or empty vector lentivirus for 48 hours were subcutaneously injected into the right axilla of 4-week-old BALB/c nude mice, respectively, as shRNA-SNAP25 transplantation group, shRNA-VAMP2 transplantation group and empty vector transplantation group (5 mice in each group); on 28 th day of injection, immunohistochemical staining was used to detect the Ki-67 expression in the LN229 cell-transplanted tumor tissues. Results:(1) A total of 1,473 DEGs were screened, of which 880 were upregulated and 593 were downregulated. WGCNA indicated that DEGs were divided into 5 modules (greenish-blue, blue, black, brown and gray ones), among which the greenish-blue module was significantly negatively correlated with GBM ( r=-0.700, P<0.001); GSEA analysis showed that the greenish-blue module mainly involved Notch signaling pathway, and PPI network analysis identified SNAP25 and VAMP2 as hub genes. (2) Immunohistochemical staining results showed that expressions of SNAP25 and VAMP2 in GBM tissue were significantly lower than those in normal brain tissue ( P<0.05). (3) Compared with those in the empty vector group, the SNAP25 mRNA and protein expressions in LN229 and U87 cells of the shRNA-SNAP25 group were statistically decreased ( P<0.05). Compared with those in the empty vector group, the VAMP2 mRNA and protein expressions in LN229 and U87 cells of the shRNA-VAMP2 group were significantly decreased ( P<0.05). Compared with the empty vector group, the LN229 and U87 cells in the shRNA-SNAP25 group and shRNA-VAMP2 group had significantly increased colony formation number, Ki-67 expression and invasive cell number ( P<0.05). Compared with the empty vector group, LN229 cells in the shRNA-SNAP25 group and shRNA-VAMP2 group had statistically increased Notch1, HEY1, and HES1 protein expressions ( P<0.05). Immunohistochemical staining results showed that compared with that in the empty vector group (1.00±0.00), the Ki-67 expression in LN229 cell-transplanted tumor tissues of the shRNA-SNAP25 group and shRNA-VAMP2 group was statistically increased (1.41±0.05, 1.40±0.09, P<0.05). Conclusion:Hub genes SNAP25 and VAMP2 may negatively regulate the malignant biological behavior of GBM through Notch pathway, which might be the new candidate targets for GBM precise treatment.

Objective:Glioblastoma (GBM) is a primary malignant tumor in the central nervous system, whose tissue heterogeneity and invasive growth characteristics lead to a very poor prognosis for patients. Hub genes in GBM are screened by bioinformatics analysis; expressions of Hub genes in human GBM tissue, and their effects on GBM biological behaviors and Notch signaling pathway proteins are explored, and their regulatory roles in GBM proliferation in xenograft tumor model mice are evaluated.Methods:(1) The gene expression profiles of GBM tissue and normal brain tissue from the Gene Expression Omnibus (GEO) database were obtained; differentially expressed genes (DEGs) were screened, and gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses were conducted on DEGs. A co-expression network was constructed using weighted gene co-expression network analysis (WGCNA) to identify modules significantly related to GBM. Gene set enrichment analysis (GSEA) on the related module genes was performed; STRING database (version 12.0) was used to construct protein-protein interaction (PPI) network and screen the hub genes. (2) Normal brain tissue samples from 8 patients with epilepsy and GBM tissue samples from 10 patients with GBM who underwent surgical resection in Department of Neurosurgery, He'nan Provincial People's Hospital from January 1, 2022 to December 1, 2023 were collected; expressions of synaptosomal-associated protein 25 (SNAP25) and vesicle-associated membrane protein 2 (VAMP2) were detected by immunohistochemical staining and Western blotting. (3) LN229 and U87 cells were routinely cultured in vitro and divided into shRNA-SNAP25/shRNA-VAMP2 group and empty vector group, and then, cells in each group were transfected with shRNA-SNAP25/shRNA-VAMP2 lentivirus or empty vector lentivirus, respectively; 24 hours after transfection, SNAP25 and VAMP2 mRNA and protein expressions in the 2 groups were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and Western blotting. LN229 and U87 cells were routinely cultured in vitro and divided into shRNA-SNAP25 group, shRNA-VAMP2 group and empty vector group; cells in each group were transfected with shRNA-SNAP25 lentivirus, shRNA-VAMP2 lentivirus or empty vector lentivirus, respectively; 48 hours after transfection, proliferation was detected by clone formation assay, proliferating cell nuclear antigen Ki-67 expression was detected by immunofluorescent staining, invasion was detected by Transwell invasion assay, and Notch1, HEY1 and HES1 protein expressions in Notch signaling pathway of LN229 cells were detected by Western blotting. (4) LN229 cells transfected with shRNA-SNAP25 lentivirus, shRNA-VAMP2 lentivirus or empty vector lentivirus for 48 hours were subcutaneously injected into the right axilla of 4-week-old BALB/c nude mice, respectively, as shRNA-SNAP25 transplantation group, shRNA-VAMP2 transplantation group and empty vector transplantation group (5 mice in each group); on 28 th day of injection, immunohistochemical staining was used to detect the Ki-67 expression in the LN229 cell-transplanted tumor tissues. Results:(1) A total of 1,473 DEGs were screened, of which 880 were upregulated and 593 were downregulated. WGCNA indicated that DEGs were divided into 5 modules (greenish-blue, blue, black, brown and gray ones), among which the greenish-blue module was significantly negatively correlated with GBM ( r=-0.700, P<0.001); GSEA analysis showed that the greenish-blue module mainly involved Notch signaling pathway, and PPI network analysis identified SNAP25 and VAMP2 as hub genes. (2) Immunohistochemical staining results showed that expressions of SNAP25 and VAMP2 in GBM tissue were significantly lower than those in normal brain tissue ( P<0.05). (3) Compared with those in the empty vector group, the SNAP25 mRNA and protein expressions in LN229 and U87 cells of the shRNA-SNAP25 group were statistically decreased ( P<0.05). Compared with those in the empty vector group, the VAMP2 mRNA and protein expressions in LN229 and U87 cells of the shRNA-VAMP2 group were significantly decreased ( P<0.05). Compared with the empty vector group, the LN229 and U87 cells in the shRNA-SNAP25 group and shRNA-VAMP2 group had significantly increased colony formation number, Ki-67 expression and invasive cell number ( P<0.05). Compared with the empty vector group, LN229 cells in the shRNA-SNAP25 group and shRNA-VAMP2 group had statistically increased Notch1, HEY1, and HES1 protein expressions ( P<0.05). Immunohistochemical staining results showed that compared with that in the empty vector group (1.00±0.00), the Ki-67 expression in LN229 cell-transplanted tumor tissues of the shRNA-SNAP25 group and shRNA-VAMP2 group was statistically increased (1.41±0.05, 1.40±0.09, P<0.05). Conclusion:Hub genes SNAP25 and VAMP2 may negatively regulate the malignant biological behavior of GBM through Notch pathway, which might be the new candidate targets for GBM precise treatment.

Objective:To explore the risk factors of pressure injury (PI) in postoperative coma patients with severe brain injury (SBI) .Methods:From June 2018 to April 2022, 300 postoperative coma patients with SBI admitted to Henan Provincial People's Hospital were selected as the study subject by convenience sampling. Patients were divided into PI group ( n=35) and non-PI group ( n=265) based on whether PI occurred 7 days after surgery, and the clinical data of the two groups of patients were compared. Logistic regression was used to analyze the risk factors for postoperative PI in coma patients with SBI. Results:Among 300 postoperative coma patients with SBI, the incidence of PI during hospitalization was 11.67% (35/300), with a total of 54 occurrences of PI. Logistic regression analysis showed that age, diabetes, malnutrition, urinary and fecal incontinence, respiratory mode and postoperative hyperthermia were all the influencing factors of PI in coma patients with SBI after surgery ( P<0.05) . Conclusions:Age, diabetes, malnutrition, urinary and fecal incontinence, mechanical ventilation and postoperative high fever are all risk factors for PI in coma patients with SBI. Nurses should actively evaluate the risk of PI and provide targeted preventive measures.

Traumatic cerebrospinal fluid leakage commonly presents in traumatic brain injury patients, and it may lead to complications such as meningitis, ventriculitis, brain abscess, subdural hematoma or tension pneumocephalus. When misdiagnosed or inappropriately treated, traumatic cerebrospinal fluid leakage may result in severe complications and may be life-threatening. Some traumatic cerebrospinal fluid leakage has concealed manifestations and is prone to misdiagnosis. Due to different sites and mechanisms of trauma and degree of cerebrospinal fluid leak, treatments for traumatic cerebrospinal fluid leakage varies greatly. Hence, the Craniocerebral Trauma Professional Group of Neurosurgery Branch of Chinese Medical Association and the Neurological Injury Professional Group of Trauma Branch of Chinese Medical Association organized relevant experts to formulate the " Chinese expert consensus on the diagnosis and treatment of traumatic cerebrospinal fluid leakage in adults ( version 2023)" based on existing clinical evidence and experience. The consensus consisted of 16 recommendations, covering the leakage diagnosis, localization, treatments, and intracranial infection prevention, so as to standardize the diagnosis and treatment of traumatic cerebrospinal fluid leakage and improve the overall prognosis of the patients.

Objective:To effectively express the receptor binding domain (RBD) of SARS-CoV-2 spike protein in Pichia pastoris and to evaluate its immunogenicity. Methods:The gene encoding the RBD protein was synthesized and cloned into the pPICZαA plasmid. After linearization, the plasmid was transferred and integrated into the genome of Pichia pastoris. The expressed RBD protein in culture supernatant was analyzed by Western blot and Biolayer interferometry. After screening, a single clone expressing the RBD protein was selected. The high-level expression of RBD protein was achieved by optimizing the fermentation process, including the salt concentration adjusting of the medium and induction condition optimization (pH, temperature and duration). The immunogenicity of the expressed RBD protein was evaluated in a mouse model. Results:A single clone with a high expression level of RBD protein was obtained and named RBD-X33. The expression level of RBD protein in the fermentation supernatant reached up to 240 mg/L after optimization of the induction condition (HBSM medium, pH=6.5±0.3, 22℃ and 120 h). In the mouse experiment, the recombinant RBD protein was formulated with Alum+ CpG dual adjuvant and injected into mice. The binding IgG antibody levels were up to 2.7×10 6 tested by ELISA and the neutralizing antibody levels were up to 726.8 tested by live virus neutralizing antibody assay (prototype). Conclusions:The RBD protein could be efficiently expressed in Pichia pastoris and induce stronger immune response in animals. This study suggested that the recombinant SARS-CoV-2 RBD protein expressed in Pichia pastoris could serve as a candidate antigen in the development of SARS-CoV-2 vaccine.

Myasthenia gravis(MG)is an autoimmune disease, and local or systemic muscle weakness is the main symptom.The main auxiliary tests are serological, pharmacological and electrophysiological tests.Autoantibodies are the most important biomarkers to guide the diagnosis of myasthenia gravis.The clinical features and autoantibodies of myasthenia gravis in children were different from those in adults.Therefore, this paper summarizes the clinical significance of the main autoantibodies acetylcholine receptor(AChR)antibody, muscle specific kinase(MuSK)antibody, low density lipoprotein receptor associated protein 4(LRP-4)antibody and other related antibodies in children with myasthenia gravis, which has certain guiding significance for the diagnosis and individualized treatment of myasthenia gravis.

Objective:To investigate the efficacy and safety of neuroendoscopy in the treatment of non-acute traumatic intracranial hematoma.Methods:Thirty-six patients with non-acute traumatic intracranial hematoma, admitted to our hospital from June 2018 to December 2019, were chosen in our study. These patients accepted small-bone window craniotomy and straight incision, or removal of intracranial hematoma by neuroendoscopy. The clinical data of these patients were retrospectively analyzed. Pain numerical rating scale (NRS) was used to assess degrees of pain in 22 patients with headache one d before surgery and three d after surgery. The neurological functions after treatment were evaluated by activity of daily living (ADL) evaluation criteria one d before surgery and 7 d after surgery.Results:All 36 patients were cured and discharged from hospital, and no death was noted; length of hospital stays was (6.7±1.1) d. No secondary hemorrhage re-craniotomy was needed, no postoperative complications such as cerebrospinal fluid leakage were noted, and no re-injection of urokinase was needed to melt the hematoma. As compared with the preoperative NSR scores (7.82±1.097), the postoperative NSR scores of 22 headache patients were significantly decreased (1.05±0.653, P<0.05). In these 36 patients, preoperative ADL level I was noted in 8 patients, level II in 14 patients, level III in 12 patients, and level IV in 2 patients. Postoperative nerve function in 30 patients were fully recovered, with ADL level I; and 6 patients had mild symptom of dizziness, with ADL level II. Conclusion:Non-acute traumatic intracranial hematoma treated by neuroendoscopy enjoys good curative effect, less surgical trauma, short hospital stays and high safety.

Objective To discuss the strategies and efficacies of surgical treatment in medulla oblongata cavernous malformations.Methods The clinical and follow-up data of 9 patients with medulla oblongata cavernous malformations,admitted to our hospital from October 201 1 to February 2015,were retrospectively analyzed.Five patients underwent surgical treatment:posterior midline transcerebellomedullary fissure approach was used in 4,far-lateral transcondylar approach in one,and all cavernous malformations were excised completely.Conservative treatment and regular observation were done in 4 patients.Follow-up was performed for 10-40 months.Results Symptoms improved after surgery in 4 patients;postoperative hemiplegia appeared in one,and at the end of follow-up,the muscle strength of hemiplegic limbs recovered to level Ⅳ.Minor re-hemorrhage was noted in one of the 4 patients with conservative treatment at the end of the follow-up,and no changes were noted in the rest patients.Karnofsky Performance Scale scores were 82.2 ±6.7,which were significantly increased as compared with those before treatment (73.3±11.2,P＜0.05).Conclusion After strict preoperative evaluation,surgical treatment is a feasible way,enjoying good prognosis in medulla oblongata cavernous malformations;but,patients with deep site,small malformations or no obvious bleeding symptoms should not be operated.

The expression of paired immunoglobulin-like receptor B (PirB) in normal and injured spinal cord of rats was investigated. The SD rat hemi-sectioned spinal cord injury (SCI) model was established. Before and 1, 3, 7, 10 days after SCI, the spinal cord tissues were harvested, and Western blot and immunohistochemistry were used to examine the expression and location of PirB. The results showed that the expression level of PirB in the normal spinal cord of SD rats was low. At the first day after SCI, the expression of PirB was obviously increased, and that in the injured spinal cord from the first day to the 10th day was significantly higher than in the normal spinal cord. The positive expression of PirB in neurons from different regions of gray matter of the injured spinal cord was seen. It was concluded that the expression of PirB in the normal spinal cord of rats was low. The expression of PirB in SCI was significantly increased till at least the 10th day.