1.ATP1A1 gene silencing inhibits the invasion of human glioma cells U251 in vitro
Hong CHEN ; Song CHEN ; Gang HUO
Basic & Clinical Medicine 2017;37(4):506-511
Objective To investigate the effect and mechanism of silencing ATP1A1 gene on invasion ability of human U251 glioma cells.Methods The human U251 glioma cells were infected with lentivirus expressing shRNA-ATP1A1.The mRNA and protein expression of ATP1A1 in U251 glioma cells was detected by RT-qPCR and Western blot,respectively.The proliferation of U251 glioma cells was determined by MTT assay.The migration and invasion ability were detected by cell scratch test and Transwell chamber.The protein expression of matrix metallo proteinase 2 (MMP-2) and MMP-9 in U251 glioma cells were detected by Western blot.Results The mRNA and protein expression of ATP1A1 in the silence group were significantly inhibited,The ability of proliferation, migration and invasion were also significantly inhibited (P<0.05),The expression of MMP-2 and MMP-9 were also significantly reduced,there was a significant difference(P<0.05).Conclusions RNA interference targeting ATP1A1 gene can significantly inhibit the proliferation, migration and invasion of glioma U251 cells.The mechanism might be related to the down-reguLation of MMP-9 and MMP-2.ATP1A1 can be used as a potential target for the treatment of glioma.
2.Effects of ischemic postconditioning on NF-κB and ICAM-1 expression during lung ischemia-reperfusion injury in a dog model of CPB
Lu YOU ; Song CHEN ; Hong ZHANG
Chinese Journal of Anesthesiology 2015;35(7):868-871
Objective To evaluate the effects of ischemic postconditioning on nuclear factor kappa B (NF-κB) and intercellular adhesion molecule-1 (ICAM-1) expression during lung ischemia-reperfusion (I/R) injury in a dog model of cardiopulmonary bypass (CPB), and further investigate the pulmonary protection induced by ischemic postconditioning and the underlying mechanism.Methods Twelve adult mongrel dogs of both sexes, weighing 12-15 kg, were randomly divided into either CPB group or ischemic postconditioning group (IPO group) using a random number table, with 6 dogs in each group.CPB was established after the chest was opened in dogs anesthetized with pentobarbital sodium.Ischemic postconditioning was induced by 2 cycles of 5 min reperfusion followed by 5 min ischemia immediately after occlusion of the left pulmonary artery was released in group IPO.Before CPB (T1), before occlusion of the artery was released (T2) , and at 2 h after termination of CPB, lung specimens were obtained for examination of pathological changes which were scored (with light microscope) and for determination of the expression of NFκB and ICAM-1 (using Western blot) and wet/dry lung weight ratio (W/D ratio) in left lung tissues.Blood samples were collected from femoral arteries at T1 and T3 for blood gas analysis, and oxygenation index (OI), respiration index (RI) and dynamic lung compliance (Cdyn) were calculated.Results OI and Cdyn were significantly decreased, and RI was increased at T3 , and W/D ratio, pathological scores, and expression of NF-κB and ICAM-1 were increased at T2,3 than at T1 in the two groups.W/D ratio, pathological scores, and expression of NF-κB and ICAM-1 were significantly higher at T3 than at T2 in the two groups.Compared with group CPB, OI and Cdyn were significantly increased, and R1, W/D ratio, pathological scores, and expression of NF-κB and ICAM-1 were decreased at T3 in group IPO.Conclusion Ischemic postconditioning up-regulates the expression of ICAM-1 through inhibiting NF-κB activity, thus reducing lung I/R injury induced by CPB and improving the lung function in dogs.
3.Mobile Vehicle Exhaust Exposure and Impacts of Cardiovascular System in Middle and Old Aged People:a panel Study in a Community of Guangzhou
Boning CHEN ; Hong SONG ; Ganjing WEN
Journal of Environment and Health 2007;0(09):-
Objective To explored the effects of vehicle exhaust on the cardiovascular system in a panel of 32 middle and old aged people,to discover the sensitive effect indicators of vehicle exhaust. Methods Thirty-two middle and old aged people living near a main road were followed-up,The health-related indicators and individual time-weighted outdoor exposure concentration of air pollutants were determined;The mixed model was used to evaluate association between vehicle exhaust and health indicators of cardiovascular system. Results In single-pollutant models,TNF-? was positively associated with PM10,SO2,NO2 and FVC,IL-6 was negative. The indicators of heart rate variability(HRV) in SDNN(?:-0.57,SE:0.18) ,SDNN index(?:-0.74,SE:0.20) had a strong association with PM10,and NO2 related indicators was SDNN index(?:-0.66,SE:0.17) . In the multi-pollutant model,after adjusting the effect of other pollutants,IL-6 had a positive correlation with PM10;And TNF-? was positively correlated to NO2,SO2;FVC had a negative correlation with PM10. Conclusion The air pollutants of this location show a vehicle exhausts characteristic,and the pollution is serious;TNF-?,IL-6,FVC,SDNN,SDNN index are closely related to exposure of the vehicle exhaust,and can be used to evaluate the effects of vehicle exhaust pollution on the cardiovascular system of the elderly.
4.Effect of Vehicle Exhaust Pollution on Lung Function and Inflammatory Factors Level in Pupils
Qiang TAN ; Hong SONG ; Suimei CHEN
Journal of Environment and Health 2007;0(09):-
Objective To explore whether pollutants exposure has a measurable impact on lung function and inflammatory factors level of healthy pupils in vehicle exhaust polluted region. Methods Primary school near the trunk road with a distance no more than 30 meters was selected,and 50 qualified pupils were chosen. A time-series panel study was conducted with these pupils,and the following consecutive five days'study of vehicle exhaust pollution level,3-days’personal exposure measurement and its health effects were carried out in May and November in 2008. Results The main pollutant in this region was motor vehicle exhaust. Compared to May,the value of FVC% declined significantly in November(P=0.02) . And the reduction of the value of FVC% was negatively associated with an increase in NO2 concentrations(P=0.04) . The levels of IL-2,IL-6,IL-8 became lower in November compared with May,and were negatively associated with personal exposure with IL-6 with statistically significant difference(?PM10=-0.62,?NO2=-0.62,?O3=-0.64,?SO2=-0.63) . The levels of TNF-? and IL-4 were positively associated with personal exposure,in which PM10 has the largest regression coefficient with TNF-?(?=0.65,P
5.Hemagglutinin A of Porphyromonas gingivalis function as adhesion and invasion in human gingival epithelial cells
Hong SONG ; Qin MA ; Yongjin CHEN
Journal of Practical Stomatology 2000;0(05):-
Objective: To determine if the hemagglutinin A (HagA) of Porphyromonas gingivalis could be involved in the adhesion and invasion in human gingival epithelial cells (HGEC). Methods:P. gingivalis 381 hagA mutant was constructed by conjugation method. The whole length of hagA gene was cloned into pYA292 in Salmonella typhimurium x4072 (S. typhimurium-hagA). The strains were used to test their ability of adhesion and invasion into HGEC using a standard antibiotic protection assay. S. typhimurium x4072 strains containing empty vectors were used as negative control. HagA expression in S. typhimurium-hagA was confirmed by Western blot. Results:Although there were no significant differences between P. gingivalis 381 hagA mutant and wild type in adhesion and invasion into HGEC, the adhesion values of S. typhimurium-hagA to HGEC were increased by 3 times compared to their respective controls, while the invasion ability of S. typhimurium-hagA was 4 times greater than that of the negative controls. Conclusion: These results suggest that HagA may participate in P. gingivalis adhesion and invasion into HGEC.
7.Expression and location of intracellular tissue factor in atherosclerosis stable plaque of ApoE(-/-) mice.
Jun, LI ; Tao, CHEN ; Dingmiao, WANG ; Yifeng, SONG ; Mei, HONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):457-61
In the ApoE(-/-) mouse model of atherosclerosis (AS) stable plaque, the expression and location of intracellular tissue factor (TF) in the cellular components of AS stable plaque were investigated in order to explore the cellular mechanism of AS thrombosis. Pathological changes of the stable plaque were observed under a microscope. The expression of TF protein was examined in aortic stable plaque of mice by using immunohistochemistry. Color image planimetric system was used to analyze the histological components of the stable plaque and the TF distribution. Under the confocal microscope, the intracellular TF location in the stable plaque of mice was observed. The results showed the cellular area was the major part of stable plaque (67.36%+/-6.52%, P<0.01). The percentage of total area occupied by cellular area was significantly larger than atheromatous gruel and acellular area (P<0.01). Macrophages and smooth muscle cells (SMC) were major cells in the cellular area. The percentage of total area occupied by SMC was significantly larger than by macrophages (P<0.01). Multiple linear regression analysis showed there was a positive correlation between TF area and SMC area (r=0.616, P=0.008), and no correlation was found between TF area and macrophage area (r=0.437, P=0.08). Pictures of color image planimetric analysis of TF and SMC were merged to highlight areas with co-localization (yellow), it was concluded that the process could be a cell-mediated TF expression in the stable plaque. SMC may be the major source of TF in AS without plaque rupture.
8.Effect of GABA on expression of VEGF and MMP-9 in monocrotaline-induced pulmonary hypertension rats
Gang CHEN ; Jun SONG ; Hong XIONG ; Lujie ZHANG ; Jinquan LV
Chongqing Medicine 2014;(30):4043-4045
Objective To study the therapeutical effect of γ-aminobutyric acid on monocrotaline(MCT )induced pulmonary hy-pertension rats ,and to elucidate the expression of VEGF and MMP-9 .Methods Thirty SD rats were randomly divided into 3 groups :a normal control group(control group) ,a MCT-induced pulmonary hypertension group(model control group) ,and anγ-ami-nobutyric acid treatment group(treatment group) .The mean right ventricular pressure(mRVP)were detected ,the right ventricular hypertrophy index(RVHI)were measured ,WT% and WA% were evaluated ,and the expression of VEGF mRNA in the lung tissue and MMP-9 were detected wtih FQ-PCR and immunohistochemical staining method respectively .Results mRVP ,RVHl ,WT% , WA% and the expression of MMP-9 and VEGF mRNA of treatment group were lower than those in the model control group(P<0 .05) ,but higher than the control group(P<0 .05) .Conclusion GABA has a therapeutic effect on pulmonary hypertension rats through regulating the expression of VEGF mRNA and MMP-9 protein .
9.Changes of serum osteoprotegerin and bone mineral density in diabetic nephropathy patients
Defang HUANG ; Gaoxiang CHEN ; Song ZHANG ; Fang ZHANG ; Hong CAO
Chinese Journal of Tissue Engineering Research 2010;14(2):351-354
BACKGROUND: At present, the relationship between diabetic nephropathy and osteoprotagerin remains poorly understood. OBJECTIVE: To investigate the correlation between bone mineral density (BMD) and serum osteoprotagerin changes in patients with 2 type diabetic nephropathy. METHODS: Totally 104 patients with 2 type diabetes were divided following 5 groups according to glomerular filtration rate: simple diabetes, mild renal injury, moderate renal injury, severe renal injury, and renal failure groups. Additional 20 healthy people were selected as the control group. The level of serum osteoprotegerin was measured by ELISA. Meantime, levels of serum calcium, phosphonium, alkaline phosphatase, creatinine, urea nitrogen, and glycosylatad hemoglobin were measured by. the automatic biochemistry analyzer. The bone mineral density of entopic L_(2-4) was determined by dual X-ray bone density equipment. The whole data was analyzed by multiple regression correlation analysis. RESULTS AND CONCLUSION: The level of serum osteoprotegerin in patients with diabetic nephropathy was obviously greater than that of the healthy people (P < 0.05), but BMD of the mild renal injury, moderate renal injury, severe renal injury, and renal failure groups was obviously lower than that of the healthy people (P < 0.05). Generally, the worse renal function accompanied by higher osteoprotegerin level, and lower BMD. There was a negative correlation between ostaoprotegerin level and BMD in patients with diabetic nephropathy (r=-0.497, P < 0.01). However, the relationships between osteoprotegedn level and diabetic duration (r=0.566, P < 0.01), serum creatinine level (r=0.772, P < 0.01), serum urea nitrogen level (r=0.708, P < 0.01), serum phosphonium level (r=0.329, P < 0.01), or serum intact parathyroid hormone level (F=0.702, P < 0.01) were positive. Meantime, the serum phosphonium level had negative correlation to serum calcium level (r=-0.505, P < 0.01). it demonstrated that when the renal function got worse in diabetic nephropathy patients, the serum osteoprotegerin level was increasing accompanied by BMD decreasing. The osteoprotegerin level presents a negative correlation to BMD and serum calcium level, but positive to diabetic duration, serum creatinine, serum urea nitrogen, serum phosphonium and serum intact parathyroid hormone levels.
10.The association between snoRNA profiles and prognosis in patients with gastric cancer
Lu HAN ; Fengju SONG ; Yubei HUANG ; Hong ZHENG ; Kexin CHEN
Tianjin Medical Journal 2016;44(5):543-547
Objective To identify snoRNA, which may be related to prognosis of gastric cancer. Methods Ninetygastric cancer patients who diagnosed at Tianjin Medical University Cancer Institute and Hospital were randomly collected in this study, and their clinical data were followed up. A total of 405 snoRNA expression profiles were analyzed in 90 gastric cancer patients. Patients were classified aslow expressiongroup orhigh expressiongroup according to the median expression of each snoRNA expression, which was calculated by univariate and multivariate survival analysis. We also screened out the snoRNAs, in which patients were survived differently. Patients were classified as high, middle, or low risk groups based on the snoRNA risk score. Values of age, gender, smoking, drinking, histological differentiation (well, moderately-differentiated and poorly differentiated), clinical stage (Ⅰ+Ⅱstage andⅢ+Ⅳstage), tumor size (<5 cm and≥5 cm), tumor location (upper 1/3 and others) and snoRNA risk score (high, middle, and low risk group) were assessed by multivariate Cox analysis. Results There were significant differences in overall survival and (or) progression-free survival rates in 19 patients with high and low snoRNAs expressions (P<0.05). Results of multivariate Cox analysis showed that patients with high expression of ACA61,ACA27 and U36A showed a higher overall survival and progression-free survival rates, while patients with high expression of ENSG00000206898 showed a lower overall survival and progression-free survival survival rates (P<0.01). SnoRNA risk score is an independent prognostic factor for patients with gastric cancer. Compared with low risk group, patients in middle risk group and in high risk group showed a shorter overall survival and progression-free survival rates (P<0.001). Conclusion The expressions of ACA61, ACA27, U36A and ENSG00000206898 are independent prognostic factors of gastric cancer. Low expressions of the first three indexes and high expressions of the last one predict a bad prognosis.