1.Analysis of risk factors and body composition in elderly patients with type 2 diabetes complicated nonalcoholic fatty liver disease
Pengju LIU ; Fang MA ; Huiping LOU ; Chunwei DU ; Xin TAO
Chinese Journal of Geriatrics 2010;29(2):111-114
Objective To investigate the probable risk factors for type 2 diabetic patients complicated nonalcoholic fatty liver disease (NFLD)in elderly, through comparing the body composition, serum lipid profile, incidences of abdominal obesity and metabolic syndrome (MS) between elderly type 2 diabetic patients with and without NFLD. Methods The enrolled elderly type 2 diabetic patients were divided into NFLD group (n=83) and non-NFLD group (n=85). Their clinical data including body composition, serum lipid profile, incidences of abdominal obesity and MS were analyzed retrospectively and compared. Results Compared with non-NFLD group, the BMI [(26.9±2.5) kg/m~2 vs. (24.1±2.5) kg/m~2, P=0.000], waist-hip ratios (WHR) ((0.92±0.07) vs. (0.87±0.06), P=0.000], total body fat percentage [(29.6%±6.6%) vs. (25.3%±5.5%),P=0.000], abdominal fat [(11.0±2.5) kg vs. (8.7±2.3) kg, P=0.000], visceral fat [(3.0±0.7) kg vs. (2.3±0.6)kg, P=0.000], visceral fat area [(97.6±22.2) cm~2 vs. (75.5±21.1) cm~2,P=0. 000], serum triglyceride [(1.98±0.94) mmol/L vs. (1.22±0.61) mmol/L, P=0.000]were all increased, while serum HDL [(1.23±0.32) mmol/L vs. (1.40±0.37) mmol/L, P=0.002]was decreased in NFLD group. The incidences of over-body fat (68.7% vs. 36.5%, P=0. 000),dyslipidemia (47.0% vs. 21.2%, P=0. 000), abdominal obesity (69.9% vs. 43.5%, P=0.001) and MS (49.4% vs. 9.6%, P=0.000) were obviously increased. But there were no statistical differences in serum TC [(4.93±0.94) mmol/L vs. (4. 73±1.07) mmol/L, P=0.219]and LDL [(3.23±0.80) mmol/L vs. (3. 07±0.89) mmol/L, P=0. 229]between the two groups. Logistic regression showed that high BMI (β=1.268, P=0.000, OR=3.56), over-total body fat percentage (β=0.902, P=0.023, OR=2.47)and the existence of MS (β=1. 664, P=0. 000, OR=5.28) were related to elderly type 2 diabetic patients complicated NFLD. Conclusions The high BMI, over-total body fat percentage are related to elderly type 2 diabetic patients complicated NFLD, and NFLD is probably one of components of metabolic syndrome.
2.Effects of EGB on hippocampus CA1 region Caspase-9P35 activity in dementia rats induced by Dgal
Li SHAO ; Pengju MA ; Deqin GENG ; Nuan WANG ; Quansheng HUANG ; Jing BAO ; Ailing JI
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(10):884-886
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.
3.Effects of three kinds of nanoparticles on proliferation and apoptosis of esophageal squamous carcinoma cells
Pengli HAN ; Lei SUN ; Pengju LYU ; Fenfen GONG ; Chao MA ; Guo CHEN ; Yiran ZHU ; Tian XIA ; Wei CAO
Chinese Pharmacological Bulletin 2016;32(6):789-794
Aim TostudytheeffectsofCuO,ZnOand TiO2 nanoparticles on the viability and metastatic po-tential of EC-9706 and EC-109 esophageal squamous carcinomacelllineinvitro.Methods Characteristics of CuO,ZnO and TiO2 nanoparticles were detected u-sing transmission electron microscope (TEM)and dy-namic light scattering (DLS ).EC-9706 and EC-109 cells were treated with different concentrations of CuO, ZnO and TiO2 (5 ~80 mg · L-1 ).The cell prolifera-tion was analyzed by MTT assay.The cell cycle and apoptotic rates were determined by flow cytometry (FCM).The cell invasion was assayed in Transwell chambers.The expression of Bcl-2 and caspase-3 pro-tein in cells was detected by Western blot method.Re-sults CuO,ZnOandTiO2nanoparticleswerespheri-cal with primary particle size 12,20. 6,12 nm.The particles were agglomerated in water and cell culture medium with negative charge.CuO and ZnO nanoparti-cles induced decreases in EC-9706 and EC-109 cell vi-ability dose-dependently.After exposed to increasing concentrations of CuO and ZnO nanoparticles,the cell cycle analysis revealed a decreasing proportion of cells in G2/Mand S phase,and up-regulation of the cells in G0/G1 phase.Apoptotic cells also increased along with decreased cell invasion upon CuO and ZnO treatment. Nanoparticles treatment after 48 h, the activated caspase-3 expression quantity increased significantly and the Bcl-2 expression quantity decreased obviously (P<0. 05 )compared with control group.TiO2 nanop-articles had no obvious effect on the EC-9706 and EC-109 cell proliferation,cell cycle,apoptosis and inva-sion.Conclusion ComparedwithTiO2,CuOand ZnO nanoparticles can inhibit EC-9706 and EC-109 cell viability and metastatic potential,the mechanism of action involves cell cycle arrest in G0/G1 phase and apoptosis.These findings can help the development of nanoparticles as anti-cancer therapeutics for esophageal cancer.
4.Effect of soluble amyloid precursor protein α on nerve cell apoptosis and neurological function in subarachnoid hemorrhage rats
Haigang CHANG ; Lei HUI ; Pengju MA ; Xiangsheng LI ; Ruihua LIU ; Baozhe JIN
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2018;20(6):639-641
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.
5.Sodium valproate reduces high-sensitivity C-reactive protein and attenuates cerebrovascular spasm damage in rats after subarachnoid hemorrhage
Haigang CHANG ; Baozhe JIN ; Pengju MA ; Guojun GAO ; Lei HUI
Chinese Journal of Neuromedicine 2016;15(2):136-139
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.
6.Complement C3a receptor 1 high-expression progresses malignancy in glioma through nuclear factor-kappa B signaling pathway
Xiansheng LI ; Yanjuan MA ; Pengju MA ; Zhijie YIN ; Fuguang LI ; Weilong YANG ; Lei HUI
Chinese Journal of Neuromedicine 2022;21(12):1195-1202
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.
7.Relationship between normal serum uric acid levels and nonalcoholic fatty liver disease in postmenopausal women.
Pengju LIU ; Fang MA ; Huiping LOU ; Yanning ZHU ; Yu CHEN
Chinese Journal of Hepatology 2014;22(1):53-57
OBJECTIVETo analyze the relationship between normal serum uric acid (SUA) levels and nonalcoholic fatty liver disease (NAFLD) among postmenopausal women, and determine the possible risk factors of NAFLD in this patient population.
METHODSChinese postmenopausal women who participated in the annual health check-up program from March 2009 to February 2010 were retrospectively assessed to identify individuals with SUA within normal range for study inclusion. For the total 1425 study participants, the recorded data of anthropometric parameters, metabolic factors, and serum biochemical parameters were collected. Results from abdominal ultrasonography examination were used to group participants according to presence of fatty liver. Women with fatty liver were divided into NAFLD and non-NAFLD groups. Further sub-grouping was performed according to SUA quartiles, as follows: Q1 group: less than 226.1 mumol/L); Q2 group: 226.1 mumol/L less than or equal to SUA less than 267.8 mumol/L; Q3 group: 267.8 mumol/Lless than or equal to SUA less than 303.5 mumol/L); Q4 group: 303.5 mumol/Lless than or equal toSUAless than or equal to357.0 mumol/L. The independent-sample t-test was used to compare normally distributed variables between groups, and the Mann-Whitney U test was used to analyze variables with skewed distribution. Categorical variables were examined by the R * C x2 test. Binary logistic analysis was used to determine the risk factors for fatty liver and to adjust for possible confounders. The multiple non-parameter independent-sample test (Kruskal-Wallis test) was used to compare the differences of SUA levels among NAFLD groups with different disease severity.
RESULTSThe prevalence of NAFLD among Chinese postmenopausal women with normal SUA was 32.8%, with NAFLD prevalences of 20.4% (70/343) in women with Q1 SUA, 26.3% (104/395) with Q2 SUA, 35.2% (128/364) with Q3 SUA, and 51.4% (166/323) with Q4 SUA. The prevalence of fatty liver showed a significant increasing trend according to the SUA quartile (x2 = 76.470, P-trend less than 0.01). Women in the SUA Q3 and Q4 groups had significantly higher risk of fatty liver presence than women in the Q1 group (P less than 0.01 for both, with or without adjustment of confounders). Disease severity did not appear to be related to disease severity, as the SUA levels in women with mild, moderate or severe fatty liver were not significantly different (286.8+/-48.2 mumol/L vs. 277.9+/-53.0 mumol/L vs. 281.4+/-48.2 mumol/L, respectively; x2 = 3.025, P more than 0.05).
CONCLUSIONSUA levels were independently correlated with NAFLD in Chinese postmenopausal women. SUA levels in the higher quartiles of the normal range may be an independent risk factor of NAFLD.
Aged ; Female ; Humans ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; blood ; diagnosis ; Postmenopause ; Retrospective Studies ; Risk Factors ; Uric Acid ; blood
8.Role of human umbilical cord mesenchymal stem cells in the maturation and differentiation of dendritic cells in patients with severe acute pancreatitis and its mechanism in the inflammation modulation
Make LIANG ; Xuemin LI ; Lei SUN ; Baodong MA ; Pengju LYU ; Han YUE
Chinese Journal of Pancreatology 2021;21(3):201-207
Objective:To investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on the maturation and differentiation of dendritic cells (DCs) and the mechanism involved in the regulation of inflammation in patients with severe acute pancreatitis (SAP).Methods:The full-term fetal umbilical cords(about 4-5 cm) were collected from Zhengzhou Central Hospital Affiliated to Zhengzhou University after cesarean section. hUC-MSCs were isolated and cultured in primary culture. Flow cytometry was used for phenotype identification, adipogenic and osteogenic staining. 20 ml peripheral blood samples from 5 SAP patients were collected, and monocytes were isolated using lymphocyte separation solution and then induced by adding granulocyte macrophage colony-stimulating factor (GM-CSF), IL-4 and tumor necrosis factor (TNF)-α and cultured as DCs. According to different culture methods, DCs were divided into DCs group, hUC-MSCs+ DCs group and hUC-MSCs+ DCs+ NS398 group (NS398 was a specific inhibitor of COX-2, a downstream regulatory gene of NF-κB). The phenotype of DCs was detected by flow cytometry, and the levels of IL-1β, IL-lα, IL-2, IL-6 and IL-10 in the supernatant of cell culture for 24 hours were determined. The expression of toll like receptor (TLR)-4, IKKα and NF-κB-p65 were detected by Western blot.Results:The hUC-MSCs were successfully cultured, and their surface markers CD 90, CD 105 and CD 73 were positively expressed, and they could differentiate into adipocytes and bone cells. With the prolongation of culture time, DCs differentiated from immature to mature cells. Compared with the DCs group, the proportion of regulatory DCs (regDCs) was increased in the hUC-MSCs+ DCs group, and the marker CD 11b was significantly up regulated [(14.26±1.25)% vs (4. 87±0.58)%], CD 1a and CD 11c were significantly down regulated [(2.81±0.34)% vs (13.62±1.52)%, (3.88±0.5)% vs (11. 8±1.22)%]. All the difference were statistically significant ( P<0.05). The expression of IL-1β, INF-γ and IL-6 in culture supernatant were down regulated, but the difference was not statistically significant; The pro-inflammatory factor IL-1α was significantly decreased [(14.91±2.58)ng/L vs (30.19±7.75)ng/L], and the anti-inflammatory factor IL-10 was significantly increased [ (17.03±4.69)ng/L vs (1.83±0.14)ng/L]. The expression levels of NF-κB-p65 and TLR4 were significantly down regulated (0.74±0.02 vs 0.97±0.01, 0.89±0.01 vs 1.72±0.01), and the expression of IKKα protein was significantly up regulated (1.12±0.01 vs 0.21±0.01) in hUC-MSCs-DCs group. All the differences were statistically significant (all P value<0.05). Compared with DCs group and hUC-MSCs+ DCs group, the expression levels of NF-κB-p65 and TLR4 were significantly down regulated (0.34±0.01 vs 0.97±0.01, 0.74±0.02 and 0.14±0.01 vs 1.72±0.01, 0.89±0.01), while the expression of IKKα protein was significantly up regulated (1.68±0.01 vs 0.21±0.01, 1.12±0.01) in hUC-MSCs+ DCS+ NS398 group. All the differences were statistically significant (all P value<0.05). Conclusions:In SAP patients, hUC-MSCs can inhibit the maturation and differentiation of DCs, and induce CD 11bhigh CD 1alow CD 11clowrregDCs to participate in immune regulation, which may play an anti-inflammatory role by inhibiting the inflammatory cascade through TLR4/IKKα/NF-κB/COX-2 pathway.
9.Epidural fluid collection secondary to cranioplasty in 47 patients with traumatic brain injury after decompressive craniectomy
Haigang CHANG ; Yaxiao WANG ; Pengju MA ; Fazheng SHEN ; Zhijie YIN ; Ruihua LIU ; Yangyang WANG ; Xiang ZHOU ; Baozhe JIN
Chinese Journal of Neuromedicine 2018;17(3):248-253
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.
10.Down-regulation of miRNA-99b expression in mTOR/4E-BP1 signal pathway and invasion in glioma U251 cells
Qiankun JI ; Ke REN ; Jianhua ZHAO ; Zhijie YIN ; Pengju MA ; Ruihua LIU ; Baozhe JIN
Chinese Journal of Neuromedicine 2018;17(9):886-891
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.