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Objective:To investigate the complement C3a receptor 1 (C3AR1) expression in glioma and its mechanism in progressing malignancy.Methods:(1) The C3AR1 mRNA expression data and clinical information were obtained in 607 glioma patients from The Cancer Genome Atlas (TCGA) database and 656 glioma patients from Chinese Glioma Genome Atlas (CGGA) database; the differences in C3AR1 mRNA expression were analyzed among gliomas with different World Health Organization (WHO) grading. The overall survival and disease-free survival were compared between high and low C3AR1 mRNA expression patients obtained from TCGA database by Gene expression profiling interactive analysis (GEPIA). Gene body (GO) function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of C3AR1 related differentially expressed genes were performed by DAVID database. Correlation of C3AR1 mRNA expression with immune cell infiltration was analyzed using TIMER online website. (2) The brain tissues from 3 non-tumor patients and 9 glioma patients surgically resected in Department of Neurosurgery, First Affiliated Hospital of Xinxiang Medical University from January 2019 to September 2021 were collected; the C3AR1 protein expression was detected by Western blotting. (3) The in vitro cultured U87 and U251 cells were divided into negative control group and C3AR1 knockdown group ( C3AR1 being knocked down by lentivirus transfection); and CCK-8 assay, plate cloning assay and Transwell assay were used to detect the proliferation rate, number of colony formation and number of membrane penetrating cells. Western blotting was used to detect the nuclear factor-κB (NF-κB) signaling pathway protein expressions. Results:(1) In TCGA database, the C3AR1 mRNA expression in gliomas of WHO grading II, grading III and grading IV increased sequentially, with significant differences ( P<0.05). In CGGA database, the C3AR1 mRNA expression in glioma of WHO grading IV was statistically higher than that in gliomas of WHO grading II and grading III ( P<0.05). GEPIA showed that the overall survival and disease-free survival in the low C3AR1 mRNA expression group were statistically higher than those in the high C3AR1 mRNA expression group ( P<0.05). GO function analysis and KEGG pathway enrichment analysis revealed that C3AR1 related differentially expressed genes were more enriched in such biological processes and signaling pathways as calcium homeostasis, membrane structural valves, proton transmembrane transporter protein activity, chemokine signaling pathway and NF-κB signaling pathway. TIMER showed that C3AR1 mRNA expression in glioblastoma and low-grade glioma was positively correlated with infiltration degrees of B cells, CD4 + T cells, neutrophils, macrophages and dendritic cells, and C3AR1 mRNA expression in glioblastoma was negatively correlated with infiltration degree of CD8 + T cells ( P<0.05). (2) C3AR1 protein expression in glioma tissues was significantly higher than that in non-tumor tissues. (3) Compared with the negative control group, the C3AR1 knockdown group group had significantly lower proliferation rate, smaller numbers of colony formation and membrane penetrating cells, and lower expressions of NF-κB, phosphorylated (p)-NF-κB, p-NF-κB inhibitory protein (IκB)α, p-I-κB kinase (IKK)α and N-cadherin, and significantly higher E-cadherin expression. Conclusion:C3AR1 is highly expressed in glioma and progresses malignancy through NF-κB signaling pathway.
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Objective To identify the reasons and treatment strategies of epidural fluid collection (EFC) secondary to cranioplasty in patients with traumatic brain injury after decompressive craniectomy.Methods From June 2013 to July 2017,a retrospective analysis was performed on clinical data of 150 patients with traumatic brain injury after decompressive craniectomy in our hospital.A total of 47 patients experienced EFC following cranioplasty and 103 not.Risk factors of EFC after cranioplasty were analyzed by multiple factor Logistic regression.Results For the 47 EFC patients,32 patients had no obvious clinical symptoms and EFC was absorbed gradually through conservative therapy;15 patients had clinical symptoms,such as mental deterioration,headache,or limb weakness.EFC disappeared through vacuation in 4 patients and subcutaneous drainage in 11.The proportions of patients with skull defect＞80 cm2,dural defect and dural calcification in patients with EFC were significantly higher as compared with those without EFC (P＜0.05).Multiple factor Logistic regression analysis showed that skull defect＞80 cm2 and dural mater calcification were independent risk factors for EFC after cranioplasty.Conclusions Patients with large skull defect＞80 cm2 and dural calcification are prone to have EFC after cranioplasty.Careful evaluation of imaging data,good surgical skills and strengthening postoperative management can reduce incidence of EFC after cranioplasty.
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Objective To detect the down-regulation ofmiRNA-99b expression in cell invasion and its mechanism in human glioma cell line U251.Methods Glioma cell line U251 were routinely cultured in vitro.(1) U251 cells were divided into blank control group,negative control group and miRNA-99b inhibitor group;cells in the latter two groups were transfected with negative control sequences and miRNA-99b inhibitors,respectively;and cells in the blank control group did not give any treatment;mRNA expressions of miRNA-99b and mammalian target of rapamycin (mTOR) in U251 cells were measured by reverse transcription (RT)-PCR;the changes of mTOR,eIF4E-binding protein 1 (4E-BP1) andphosphorylated (p)-4E-BPl protein expressions in U251 cells were detected by Westem blotting;cell invasion was evaluated by Transwell assay.(2) U251 cells were divided into negative control group Ⅰ and mTOR siRNA group,and cells in the two groups were transfected with negative control sequences and mTOR siRNA,respectively;the miRNA-99b and mTOR mRNA expressions in U251 cells were measured by RT-PCR;the mTOR and p-4E-BP1 protein expressions in U251 cells were measured by Western blotting.(3) U251 cells were divided into miRNA-99b inhibitor+negative control group and miRNA-99b inhibitor+mTOR siRNA group,and cells in the two groups were transfected with miRNA-99b inhibitor+negative control sequences and miRNA-99b inhibitor+mTOR siRNA,respectively;the p-4E-BP1 protein expression in U251 cells was measured by Western blotting;cell invasion was evaluated by Transwell assay.Results (1) As compared with those in the blank control group and negative control group,the miRNA-99b rnRNA expression was significantly decreased,the mTOR mRNA and protein expressions and p-4E-BP1 protein expression were significantly increased,and the number of transmembrane cells was significantly larger in U251 cells of miRNA-99b inhibitor group (P＜0.05);there were no significant differences in 4E-BP1 protein expression among the three groups (P＞0.05).(2) As compared with those in the negative control group Ⅰ,the mTOR mRNA and protein expressions and p-4E-BP1 protein expression were significantly decreased in U251 cells of mTOR siRNA group (P＜0.05);there was no significant difference in miRNA-99b mRNA expression between the two groups (P＞0.05).(3) As compared with those in the miRNA-99b inhibitor+negative control group,the p-4E-BP1 protein expression and number of transmembrane cells were significantly decreased/smaller in U251 cells ofmiRNA-99b inhibitor+mTOR siRNA group (P＜0.05).Conclusions Down-regulation ofmiRNA-99b expression promotes glioma cell invasion,and its mechanism is related to the regulation of mTOR/4E-BP1 signaling pathway.
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Objective To study the effect of soluble amyloid precursor protein α (sAPPα) on nerve cell apoptosis and neurological function in subarachnoid hemorrhage (SAH) rats.Methods Sixty male SD rats were randomly divided into control group (n=20),SAH+saline group (n=20) and SAH+sAPPα group (n=20).A SAH model was established by injecting autologous blood into cistern magna in rats.After a SAH model was established for SAH + saline group and SAH + sAPPα group by injecting saline and sAPPα respectively into the cistern magna of rats,the apoptotic cells were detected by immunofluorescene with TUNEL staining and the neurological function was scored in 10 rats from each group on day 3 after injection of sAPPα and saline.Results The number of apoptotic cells in brain tissue was significantly greater in SAH+saline group than in control group (P＜0.05) and was significantly smaller in SAH+sAPPα group than in SAH+ saline group (P＜0.05).The neurological function score was 26.7±0.5,13.9±0.7 and 23.0±0.8 respectively in control group,SAH + saline group and SAH + sAPPα group.Conclusion sAPPα alleviates secondary damage of neurological function by inhibiting the apoptosis of nerve cells in rats after SAH and can thus improve their neurological function.
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Objective To explore the effect of sodium valproate (SV) on reducing high-sensitivity C-reactive protein (hsCRP) and attenuating cerebrovascular spasm damage in rats after subarachnoid hemorrhage (SAH).Methods Seventy-two male rats,weighting 300-400 g,were randomized to following experimental groups:sham-operated group,SAH group,SAH+saline treatment group,and SAH+SV treatment group (n=18).The SAH models in the later three groups were induced by injection of autologous blood into the cistern magna.Saline or SV (2 mg/100 g) was given to the rats in the SAH+saline treatment group and SAH+SV treatment group every day via intraperitoneal injection.Serum hsCRP level was measured on 1,3,5 and 10 day.Neurological deficit scale scores were assessed on 3 and 5 day.Results On 1,3,5 and 10 day,HsCRP level in the sham-operated group was (0.09± 0.02) mg/L,(0.09±0.02) mg/L,(0.09±0.02) mg/L and (0.09±0.01) mg/L;that in SAH group was (0.29± 0.01) mg/L,(0.32±0.02) mg/L,(0.35±0.02) mg/L and (0.32±0.02) mg/L;that in SAH+saline treatment group was (0.28±0.02) mg/L,(0.31 ±0.02) mg/L,(0.34±0.02) mg/L and (0.31 ±0.02) mg/L;that in SAH+SV treatment group was (0.15 ±0.02) mg/L,(0.21 ±0.02) mg/L,(0.24±0.02) mg/L and (0.15 ±0.03) mg/L;HsCRP level in the SAH group and SAH+saline treatment group was significantly higher than that in the sham-operated group (P＜0.05);HsCRP level in the SAH+SV treatment group was significantly increased as compared with that in the sham-operated group,but significantly decreased as compared with that in the SAH+saline treatment group and SAH group (P＜0.05).The neurobehavior scale scores on 3 and 5 day in SAH+SV treatment group (23.0±0.8 and 21.8±1.4) were significantly increased as compared with those in the SAH group (14.1±0.8 and 11.9±0.9) and SAH+saline treatment group (13.9± 0.7 and 11.1±1.4,P＜0.05);those in the SAH+SV treatment group (23.0±0.8 and 21.8±1.4) were significantly decreased as compared with that in the sham-operated group,but significantly increased as compared with that in the SAH+saline treatment group and SAH group (P＜0.05).Conclusion SV decreases the inflammatory injury by reducing the hsCRP level and improve the neurological outcome in SAH rat models.
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Objective To investigate the effect and mechanism of extract ginkgo biloba (EGb) on learning memory ability and hippocampus CA1 region Caspase-9P35 activity in dementia rats induced by D-gal.Methods 48 rats were randomly divided into six groups.The dementia model were established by injecting intraperitoneally with the D-gal and each EGb group was injected different doses of EGb simultaneously.TUNEL method was used to detect apoptosis of hippocampal neurons in the CA1 region.Immunohistochemistry and Western Blot were used to determine the expression of Caspase-9P35 in hippocampus CA1 region.Results Compared with the normal group,TUN EL-positive neurons ( (37.8 ± 1.3 ),(0.8 ± 0.2 ) ) and the activity of Caspase-9P35 ( (37.6 ±1.8 ),(6.2 ± 1.3 ) ) had significant increase in hippocampal CA1 subfield of D-gal group (P ＜ 0.01 ).Contrast to D-Gal group,TUNEL-positive neurons ( ( 17.6 ± 0.9),(9.8 ± 0.8 ),( 37.8 ± 1.3 ) ) and the activity of Caspase9P35( (28.6 ± 1.3),(25.0 ± 1.6),(37.6 ± 1.8) ) were significant decreased in EGb-M and EGb-H group (P＜0.01 ).While TUNEL-positive neurons and the activity of Caspase-9P35 had not significant difference in the therapy group than D-Gal group (P ＞ 0.05).Conclusion EGb can improve the cognitive level of the dementia rats.One of the therapeutic mechanisms may be to decrease the hippocampus CA1 region Caspase-9P35 activity.The results of the pretreatment group was more effective than the therapy group.