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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 detect the expression of micro RNA (miR)-433 and histone deacetylase 6 (HDAC6) in glioma tissues and investigate the effect of miR-433 on cell proliferation and invasion of human glioma cell line U251. Methods (1) Forty-two glioma samples, collected from patients accepted surgical resection and conformed by pathology in our hospital from January 2010 to December 2014, and 13 healthy brain tissues, collected from patients accepted surgery for craniocerebral trauma at the same time period, were used in our study; reverse transcription (RT)-PCR was used to detect the mRNA expression levels of miRNA-433 and HDAC6 in the glioma samples and brain tissues. (2) Human glioma cell line was routinely cultured and divided into blank control group, nonsense sequence control group and miRNA-433 mimics group;cells in the later two groups were transfected with nonsense sequences or miRNA-433 mimics, and cells in the blank control group did not give any treatment;the mRNA expression levels of miRNA-433, P21 and HDA C6 in these 3 groups were detected by RT-PCR;the cellular viability was measured by CCK-8 assay;flow cytometry was used to monitor the changes of cell cycle and apoptosis; cell invasion was evaluated by Transwell assay; HDAC6 protein expression was detected by Western blotting. (3) Wide-type (WT)HDAC63'-UTR and mutant type (MUT)HDAC63'-UTR luciferase report vectors were established; miR-433 mimics+WT HDAC63′-UTR and nonsense sequences+WT HDAC63'-UTR were transfected into the U251 cells, and dual-luciferase experiment was used to detect the fluorescence intensity of the cells; miR-433 mimics+MUT HDAC63'-UTR and nonsense sequences+MUT HDAC63'-UTR were transfected into the U251 cells, and dual-luciferase experiment was used to detect the fluorescence intensity of the cells. (4) U251 cells were divided into nonsense sequence control group, HDAC6 expression plasmids group and HDAC6 siRNA group, and nonsense sequences, HDAC6 expression plasmids or HDAC6 siRNA were transfected respectively; RT-PCR was used to detect the P21 and HDAC6 mRNA expressions and miRNA-433 expression; U251 cells were divided into miR-433 mimics group and miR-433 mimics+HDAC6 expression plasmids group, and miR-433 mimics or miR-433 mimics+HDAC6 expression plasmids were transfected, respectively, and one-5 d after that, CCK-8 was used to detect the cellular viability. Results (1) The miRNA-433 expressions gradually increased and HDA C6 mRNA expressions gradually decreased in the high-grade gliomas, low-grade gliomas and normal brain tissues, and significant differences were noted among each two groups (P<0.05);the miRNA-433 expression was negatively correlated with HDA C6 mRNA expression in the glioma tissues (r=0.829, P=0.000). (2) As compared with blank control group and nonsense sequence control group, miRNA-433 mimics group had significantly higher miRNA-433 and P21 mRNA expressions, cell percentage at G0/G1 stage, and apoptotic rate (P<0.05), and had statistically lower HDAC6 mRNA expression, cellular viability on 2-5 d of culture, number of transmembrane cells and HDAC6 protein expression (P<0.05). (3) The luciferase activity in cells from miR-433 mimics+WT HDAC63'-UTR group was significantly lower as compared with that in the nonsense sequences+WT HDAC63'-UTR group (P<0.05);the luciferase activity in cells from miR-433 mimics+MUT HDAC63'-UTR group and nonsense sequences+MUT HDAC63'-UTR group showed no significant differences (P>0.05). (4) The HDA C6 mRNA expressions were gradually increased, and P21 mRNA expressions were gradually decreased in the HDAC6 siRNA group, nonsense sequence control group, and HDAC6 expression plasmids group, with significant differences (P<0.05);on 2-5 d of culture, the cellular viability in the miR-433 mimics+HDAC6 expression plasmids group was significantly higher than that in the miR-433 mimics group (P<0.05). Conclusions The miRNA-433 expression level is low in human glioma tissues;miRNA-433 over-expression may inhibit the cell activity and promote cell apoptosis of glioma cell line U251 in vitro via inhibiting the HDAC6 expression.
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Objective To analyze the changes of serum levels ofαII-spectrin breakdown products (SBDPs)in traumatic brain inj ury (TBI)patients,and further to investigate the clinical diagnosis value of SBDPs for patients with TBI,especially with mTBI.Methods The serum levels of SBDPs were examined in 43 severe TBI (sTBI)patients,43 mild TBI (mTBI)patients and 43 healthy controls using enzyme linked immunosorbent assay (ELISA).The diagnostic usefulness of SBDPs for TBI patients were assessed by Receiver Operating Characteristic (ROC)curves analysis.Results There was no significant difference of SBDP145 among the three groups (F=1.340,P>0.05).Serum levels of SBDP120 in controls,mTBI and con-trols were 7.06±2.23,11.67±9.14 and 12.64±11.44 ng/ml,respectively.Compared with controls,serum levels of SB-DP120 were significantly higher in patients with sTBI (F=9.873,P=0.001)and mTBI (F=9.873,P=0.008),while there was no significant difference of SBDP120 between sTBI patients and mTBI patients (F=9.873,P=0.515>0.05). The area under ROC curve (AUC)of SBDP120 for TBI patients was 0.781 (95% CI:0.690~0.872,P<0.001).For mTBI patients,the area under ROC curve was 0.736 (95% CI:0.624~0.848,P<0.001).And for discriminating TBI patients with CT negative or positive,the area under ROC curve was 0.709 (95% CI:0.582~0.837,P=0.007<0.01).Conclusion The serum levels of SBDP120 were significantly increased in TBI patients,especially mTBI patients.And the serum levels of SBDP120 can be used as potential non-invasive biomarker for mTBI patients.
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Objective To compare the differences of personality in patients with depression between Uighurs and Han Chinese.Method Hospitalized depressed patients were selected including 44 cases of Uygur people,73 cases of Han people and Han people with normal control 41 cases.Using Minnesota Multiphasic Per-sonality Inventory(MMPI), Eysenck Personality Questionnai(EPQ) and Cattell's 16 Personality Factor (16PF) to make the survey.Results In MMPI : Uighur depression group's factors F,Hs, D,Hy,Pt,Pa and Sc's T score were all higher than 70,and Han depression group' s factors Hs, D,Hy,and T score of Pa were all higher than 70.Only F (76.98±16.01 vs 67.16±13.51, P＜0.01), Pt(72.09± 14.22 vs 66.82± 11.12, P＜0.05) and Sc (73.43± 13.02 vs 68.62± 11.14, P＜0.05) had statistically significant differences between the two groups.Comparing Han depression group with Han normal control group,only Pd score was not significantly different,the other nine scales were statistically significant (P＜0.01).In EPQ: comparing Uighur depression group with Han depression group,the 4 kinds of scale (extroversion, psychoticism, neuroticism and conceal) differences were not statistically significant (P＞0.05).And Han depression group compared with the Han control group, four scales were statistically significant differences (P＜0.01).In 16 PF: comparing Uighur depression group with Han depression group, only the wisdom of B (P＜0.01) and the independence of the Q2 (P＜0.05) between the two groups were statistically siguificant,other personality dimensions had no significant difference (P＞0.05).Comparing Han depression group with Han normal control group, the factors of gregariousness A, stability C, excitability D,perseverance G, boldness H, sensitivity I,skeptical L,anxiety and O,self-discipline Q3,tension Q4 (P＜0.0l) and experimental Q 1 (P＜0.05) differences were statistically significant, and the factors of Wisdom of B, aggressiveness E, fantasy M, sophisticated sex N, independence Q2 were not statistically significant (P＞0.05).Conclusion The personality model of depression between the Uygur and Han nationality has the consistency of national culture,and differences with normal people.Prompt Uygur and han depression may have a common characteristic of pathological personality model.Uighur and han ethnic differences in national culture and personality is the character of diversity,is not a Uighur and Han the pathological basis of personality of depression.