1.Regulatory fe fect of Hsf1 on PLC/PRF5 hepa toma cells proliferation
Qiying JIANG ; Zhi ZHANG ; Mingli WANG ; Yuanfang MA
Chinese Journal of Immunology 2014;(12):1601-1605
Objective:To explore the regulatory effect of Hsf 1 on PLC/PRF5 hepatoma cells proliferation.Methods: By shRNA gene silencing technology ,constructed PLC/PRF5 hepatoma cell line of Hsf 1 gene silencing.To detect the expression of Hsf 1, p53 and Rb proteins in PLC/PRF5 hepatoma cells by Western blot.The proliferation of PLC/PRF5 cell line was observed by methylthiazolyl tetrazolium assay ( MTT ) , plate clone formation assay ( PCFA ) and cell cycle assay.Results: shRNA-Hsf1 could significantly inhibit the expression of Hsf 1 in PLC/PRF5 cells.It could induce PLC/PRF5 cells stopping at G1 phase of cell cycle , inhibit cell proliferation and colonal formation;silencing Hsf1 caused up-regulation of p53 and Rb proteins expression in PLC/PRF5 cells.Conclusion: Silencing Hsf1 is involved in up-regulation of p53 and Rb proteins expression , which results in inhibiting proliferation of PLC/PRF5 hepatoma cells.
2.Effects of different dose of clopidogrel on serum CD62p level in patients with coronary heart disease
Botao ZHANG ; Xuehu ZHANG ; Lie MA ; Ping MA ; Qiaohong MA ; Mingli TAO ; Qingbin XU
Clinical Medicine of China 2014;30(4):363-366
Objective To investigate the change of serum CD62p of coronary heart disease (CHD) patients after different doses of clopidogrel administration.Methods One hundred and ninety-one patients with CHD were selected as our subjects.Of which,95 cases were with SAP and 66 cases were with non ST segment elevation acute coronary syndrome (NST-ACS).SAP patients were randomly given clopidogrel at dose of 75 mg/d or 150 mg/d and served as A and B groups.NST-ACS patients were randomly given 300 mg clopidogrel,then randomly divided into C and D groups with sequentially taking clopidogrel at dose of 75 mg/d or 150 mg/d respectively.Thirty healthy peoples were served as E group without drug intervention.Concentrations of serum CD62p were detected by Elisa before taking clopidogrel,24 h and the fifty day of after taking clopidogrel.Results (1) Before taking clopidogrel,the serum concentrations of CD62p in CHD patients were higher (A group:(7.62 ± 2.99) ng/L,B group:(8.48 ± 3.13) ng/L,C group:(9.50 ± 3.32) ng/L,D group:(10.22 ± 5.14) ng/L than that of healthy control group ((5.49 ± 1.99) ng/L,P < 0.05).The Serum CD62p levels in SAP patients were lower than that of NST-ACS patients (P < 0.05).(2)The serum concentrations of CD62p in A and B groups at before taking clopidogrel were (7.62 ±2.99) ng/L and (8.48 ±3.13) ng/L respectively,higher than that four days after taking clopidogrel ((6.79 ± 2.51) ng/L,(6.37 ± 1.80) ng/L;t =2.390,4.520;P <0.05 or P <0.01).There was no statistical significant difference between A and B groups(P >0.05).(3) In C and D groups,the serum CD62p at before taking clopidogrel were (9.50 ±3.32) ng/L and (10.22 ±5.14) ng/L,higher than that after taking clopidogrel for four days ((8.21 ± 2.62) ng/L,(8.17 ± 2.37) ng/L; t =2.084,2.157 ; P < 0.05).No significant difference was seen between C and D groups (P > 0.05).Conclusion The serum CD62p in patients with CHD was higher than that in the healthy control.Clopidogrel administration can decrease serum CD62p in CHD patients.
3.Spatial expression pattern of vascular endothelial growth factor and its receptor mRNA in the early stage of acute focal cerebral ischemia
Kangding LIU ; Ping GONG ; Xueling MA ; Jiang WU ; Yulin LI ; Mingli RAO
Chinese Journal of Tissue Engineering Research 2005;9(25):201-203
BACKGROUND: Vascular endothelial growth factor (VEGF) can accelerate neovascularization and, as a multifunctional cytokine, performs critical functions via its receptors in angiogenesis.OBJECTIVE: To investigate the expression of VEGF and its receptor FLT-1 and FLK-1 mRNA during the early stage of acute focal cerebral brain ischemia, and examine the relationship between the timing and location of their expressions.DESIGN: Randomized controlled study.SETTING: Department of Neurology of the First Hospital Affiliated to Jilin University and Teaching and Research Section of Pathology, Bethune Medical College of Jilin University.MATERIALS: This study was carried out between June 2001 and April 2002. Totally 130 adult SD rats were selected, with male and female in half, and randomly divided into normal control group (n=10), sham operation group (n=10), and cerebral ischemia group (n=110). The rats in cerebral ischemia group were further divided equally into 11 subgroups and examined at 0, 1, 2, 3, 6, 24, 48 hours and 1, 2, 3, 4 weeks after cerebral ischemia model establishment, respectively.METHODS: Permanent middle cerebral artery occlusion (MCAO) model was established in rats in cerebral ischemia group by ligation of the left common carotid artery, while the rats in the sham operation group received no artery ligation but with identical other treatments. The rats in the control group did not have any treatment. Reverse transcriptional (RT) PCR was used to detect the expression of VEGF and its receptor mRNA at different time points after ischemic model establishment, and neovascularization in the rats'brain was observed.MAIN OUTCOME MEASURES: ① Expression of VEGF and its receptor mRNA and ② neovascularization in the brain tissue at different time points of cerebral ischemia.RESULTS: Data of the 130 mice were statistically analyzed without losses.At 3, 6, 24, and 48 hours of ischemia, the number of cells positive for VEGF expression was 31.13±2.21, 43.11±2.43, 85.41±2.75 and 98.66±1.76 in each vision filed in the surrounding ischemic region, greater than the numbers in the central ischemic region at the corresponding time points (13.32±1.31, 19.40±3.22, 47.63±2.45, 57.32±3.35 in each vision field, respectively, P < 0.05). VEGF mRNA expression gradually decreased since 48 hours after model establishment till recovering the control level by 2weeks. The expression of VEGF receptor FLT-1 mRNA, determined by the number of positive cells in each vision field at 3, 6, 24, and 48 hour after the ischemia in each vision field for FLK-1 mRNA at these time points in the peripheral ischemic regions, higher than those in the central ischemic regions (P < 0.05), which recovered the control level 3 weeks after the ischemia (P < 0.05). At 48 hours and 1 week after the ischemia, the number of microvessels in each vision field was 47.2±2.11 and 199.2±3.45 in the peripheral ischemic region, significantly higher than the number in the central ischemic region (29.4±2.37 and 76.6±4.62, P < 0.05).CONCLUSION: VEGF and its receptors FLT-1 and FLK-1 mRNA are expressed in the neurons, glial cells and endothelial cells during the early stage of acute focal cerebral ischemia, and the expressions are significantly enhanced in response to ischemia, exhibiting temporal and spatial expression patterns.
4.Spatial working memory in patients with deficit and nondeficit schizophrenia
Jian WANG ; Wei DENG ; Qiang WANG ; Xiaohong MA ; Mingli LI ; Yuanyuan HAN ; Yang LIU ; Tao LI
Chinese Journal of Nervous and Mental Diseases 2013;(11):651-657
Objective To investigate the difference of spatial working memory among first-episode patients with deficit and nondeficit schizophrenia. Methods The study recruited 116 first-episode treatment-naive patients with schizo-phrenia, and 60 normal controls. Positive and Negative Syndrome Scale (PANSS) was used to assess symptoms of patients and Schedule for the Deficit Syndrome (SDS) was used to divide schizophrenia patients into deficit group (28 patients) and nondeficit group (88 patients). Spatial Working Memory (SWM) test from the Cambridge Neuropsychological Test Automat-ed Battery (CANTAB) was used to test the spatial working memory function. Results Adjusted for age, gender and years of education, there were significant differences in the performance of between errors of 4 boxes (P=0.03), between errors of 6 boxes (P=0.01), between errors of 8 boxes (P=0.03), total errors (P=0.01) and strategy (P<0.01) between deficit and nondef-icit patients, and both groups were poorer than the control group. The SWM of deficit and nondeficit patients has no corre-lation with the negative symptoms (P>0.05). Conclusion The impairment of SWM was more severe in deficit schizophrenia patients than in nondeficit patients at early stage of the disease, suggesting they are different subtypes of schizophrenia.
5.Clinical Significance of Neutrophils/Lymphocyte and Platelet/Lymphocyte Ratio of Peripheral Blood in Patients with Myasthenia Gravis
Tingting LIU ; Hu LI ; Wei MA ; Yan CHEN ; Mingli GU ; Lei ZHANG ; Anmei DENG
Journal of Modern Laboratory Medicine 2016;(1):65-68
Objective To investigate the clinical significance of neutrophils/lymphocyte ratio (NLR)and platelet/lymphocyte ratio (PLR)of peripheral blood in patients with myasthenia gravis (MG).Methods 54 patients with MG treating from Jan-uary 2013 to December 2013 in Changhai hospital and 57 healthy adults who received check-up were enrolled the study as MG group and control group.WBC count,NLR and PLR were compared between two groups and among different clinical classifications in MG patients.The area under the receiver operating characteristic (ROC)curve was performed to evaluate these indicators’diagnostic value for MG.Results The WBC count,NLR and PLR in MG group were (6.85±0.37)×109/L,2.48±0.19 and 118.79±6.38 respectively,which were significantly higher than (5.87±0.12)×109/L,1.59±0.06 and 102.01±3.45 in control group (P <0.05).The area under the ROC curve of WBC count,NLR and PLR were 0.559,0.717 and 0.581 respectively.PLR of MG patients with type Ⅲ or Ⅳ was significantly higher than that of patients with typeⅠ,Ⅱ(P <0.05).Conclusion NLR and PLR of MG patients increased significantly.The diagnostic value of NLR is higher,and PLR may be associated with the severity of disease.
6.Foxp3 overexpression in lung cancer cells suppresses immune activities of activa-ted CD4+T cells
Ruimin LIU ; Aihong ZHANG ; Weixia CHAO ; Danni SUN ; Mingli WANG ; Yuanfang MA ; Huiling BAI
Chinese Journal of Immunology 2017;33(8):1141-1145
Objective:To determine the effects of Foxp3-overexpressing lung cancer cells on activated CD4+T lymphocyte.Methods: Stable Foxp3-overexpressing lung cancer cells NCIH-1299,NCIH-hFoxp3,was generated by transfection of NCIH-1299 cells with plasmid pcDNA3-hFoxp3 mediated by Lipofectamine 2000 and by selection with G418,and validated by quantitative PCR and Western blot.The expression levels of IL-8 and IL-10 secreted by NCIH-hFoxp3 and NCIH-control were measured by ELISA.IL-2 secrection by activated human CD4+T lymphocyte which was tested after stimulation with 20% conditioned medium of NCIH-hFoxp3 and NCIH-control cells.The proliferation of activated human CD4+ T lymphocytes was assessed by MTT after coculture with NCIH-hFoxp3 cells.The adhesive ability of activated human CD4+ T lymphocytes was probed with NCIH-hFoxp3 cells by immunocytochemistry.Results: Compared with NCIH-control cells,NCIH-hFoxp3 secreted high level of IL-10 and low level of IL-8.NCIH-hFoxp3 with Foxp3 overexpression significantly suppressed the proliferation,adhesive potential and IL-2 expression by activated CD4+ T cells.Conclusion: Suppression of immune activities of activated CD4+ T cells by Foxp3 overexpression in lung cancer cells may correlate with cytokine IL-8 and IL-10,which can contribute lung cancer progression.
7.Enforced expression Foxp3 enhances proliferation and tumorigeneity of lung cancer cell
Ruimin LIU ; Weixia CHAO ; Shaolong WANG ; Mingli WANG ; Caiyun JIA ; Huiling BAI ; Yuanfang MA
Chinese Journal of Immunology 2016;32(10):1481-1484
Objective:To determine the effects of enforced expression of Foxp3 in lung cancer cell with regards to proliferation and tumorgeneity. Methods: A stable subline NCIH-hFoxp3 was established by liopfectamin-mediated pcDNA plasmid transfection carrying exogenous hFoxp3. The growth curve and secrection of IL-8 and IL-10 of NCIH-hFoxp3 were evaluated using MTT and ELISA, respectively. The in vivo tumorigeneity was assessed as well by inoculation of NCIH-hFoxp3 subcutaneously in nude mice. Results:Lung cancer cell NCIH-hFoxp3 with enforced expression of Foxp3 proliferated slowly but exihited increased in vivo tumorgeneity compared with corresponding control subline. In addition,increased expression of hFoxp3 in NCIH-hFoxp3 augmented secretion and at-tenuated secretion of IL-8 and IL-10,respectively. Conclusion:Increased expression of Foxp3 may promote progression of lung cancer cell by change of cellular microenvironment and evasion of immune surveillance.
8.Study on Separation and Purification of Total Flavones from Striga asiatica (L.) O. Ktze. With Polyamide
Yan TAO ; Song HUANG ; Mingli LUO ; Lan MA ; Xiaoping LAI ; Jiannan CHEN
Traditional Chinese Drug Research & Clinical Pharmacology 2009;20(4):380-383
Objective To study the technique condition for separating and purifying total flavones from Striga asiatica (L.) O. Ktze. by Polyamide. Methods Total flavones content, in the sample solution of Striga asiatica (L.)O. Ktze. , detected by UV spectrophotometry, is used as the index. Some technological parameters are observed by single factor observation. Results Polyamide has good absorbing effect on total flavones of Striga asiatica (L.) O. Ktze. The liquid concentration of its absorbing and separating technological condition is 1.12~2. 24 mg/mL and at the absorbing speed of 2BV/h. The elution effect of 95 % alcohol of 250 mL is the best. Conclusion This method is simple and feasi-ble, fit for separating and purifying total falvones from Striga asiatica (L.) O. Ktze.
9.Fabrication of the anti-tuberculosis controlled drug delivery system with Ti-PDA-PEG-PLGA-INH and investigation of the biological characteristics
Yunlong MA ; Litao LI ; Dan LI ; Mingli PENG ; Guanren ZHAO ; Dawei LI ; Zhanpeng LUO ; Suxi GU ; Fei YANG ; Yuanzheng MA
Chinese Journal of Orthopaedics 2016;36(11):725-734
Objective To fabricate an anti?tuberculosis controlled drug release coating with Ti?PDA?PEG?PLGA?INH and to investigate its surface characteristics, in vivo and in vitro drug release behavior, and tissue biocompatibility. Methods 4?arm?polyethylene glycol (PEG) was synthesized first. Then cover the surface of titanium (Ti) with a layer of poly dopamine (PDA) by Michael addition reaction. Use porous starch and 4?arm?PEG as a carrier, load with isoniazid (INH), then attach to the surface of titanium by casting or sol?gel dip coating methods, and then cover with a layer of poly lactic?co?glycolic acid (PLGA) by the same method, to fabricate the Ti?PDA?PEG?PLGA?INH composite coating finally. The functional group of 4?arm?PEG was charac?terized by proton nuclear resonance spectroscopy (HNMR). The surface characteristics of Ti?PDA?PEG?PLGA?INH were evaluated by scanning electron microscope (SEM), while drug release behaviors were detected by high performance liquid chromatography (HPLC) and the cumulative release rate was calculated, and carry out the antibacterial performance in vitro. The animal model of femoral condyle bone defect was established in 25 New Zealand white rabbits. Titanium rods covered with PDA?PEG?PLGA?INH coating were implanted into defect area. INH concentrations were detected by HPLC in venous blood, muscle and bone tissue at each time point postoperatively. Another 12 rabbits were randomly divided into experimental group and control group, the experi?mental group was implanted with titanium tablets and titanium rods coated with PDA?PEG?PLGA?INH in the paraspinous muscle and left femoral condyles respectively, while the control group was implanted with a blank sheet of titanium tablets and titanium rods in the same place. Hematoxylin and Eosin Staining were used to observe the biocompatibility of the composite system in vivo at 28 and 56 days postoperatively. Results Ti?PDA?PEG?PLGA?INH controlled drug release coating uniformly distributed on the surface of plates and rods, with translucent form and smooth surface. In vitro INH release kinetics exhibited a short?burst release during the first 8h, and the cumulative release of the INH was about 65%. On the 9th day, the cumulative release of the INH was about 90%, and then the release tended to be flat, and the drug release behavior in vitro continued more than 20d. In vivo release test showed that the concentration of INH in vein blood, muscle and bone tissue around the composite system was increased steadi?ly postoperatively. On about the 28th day, the concentration reached the max. However, the INH concentrations in muscle and bone tissue around the composite system were still higher than the minimum inhibitory concentration (MIC) on the 56th day. The antibacterial test in vitro showed that the titanium tablets coated with PDA?PEG?PLGA?INH formed obvious bacterial inhibition zones. The pathological results indicated that mild inflammatory reaction was seen in the 4th week postoperatively, and the reac?tive capsule formed with loose connective tissue. In the 8th week postoperatively, there's no obvious inflammation occurred, and the reactive capsule became more dense and thicker. Conclusion The study successfully fabricated the Ti?PDA?PEG?PLGA?INH anti?tuberculosis controlled drug release coating, with reasonable release behavior both in vivo and in vitro, effective antibac?terial effect of Mycobacterium tuberculosis in vitro and good tissue biocompatibility, which is a potentially effective drug delivery system for spinal tuberculosis.
10.Preliminary application of the Cambridge Neuropsychological Test Automated Battery in patients with paranoid schizophrenia and bipolar miana in China
Liqian CUI ; Zhuangfei CHEN ; Lijun JIANG ; Wei DENG ; Chaohua HUANG ; Mingli LI ; Qiang WANG ; Xiaohong MA ; Tao LI
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(5):424-427
Objective To detect the patterns of cognitive impairment between patients with paranoid schizophrenia and patients with bipolar mania by using the Cambridge Neuropsychological Test Automated Battery (CANTAB) ,and to explore research clues for finding of cognitive endophenotype in patients with paranoid schizophrenia or bipolar mania. Methods Six CANTAB subtests and the seven subtests of the Wechsler Abbreviated Scale of Intelligence (WAIS short form) were administered to 35 patients with paranoid schizophrenia and 33 patients with bipolar mania who were drug naive experiencing an acute episode, as well as 30 healthy controls. Results Patients with paranoid schizophrenia and bipolar mania demonstrated impairments in 13 of the 15 cognitive indicators in CANTAB. After controlling IQ, both patient groups remained as significantly different from normal controls in terms of search strategy(36. 8 ±3.56,37.24 ±4. 21,30. 33 ±6.24) ,between-search errors(40. 86 ± 19.97,40.24 ± 18.92,15.4 ±17.22) on the SWM test,the proportion of hits(0.54 ±0. 18,0.56 ±0.15,0.78 ± 0.17) on the RVIP test,total errors(45.26 ±36.36,46.61 ±33.32,14 ± 11.7) and EDS errors (12.43 ±9.96, 13.18 ±8.98,4.97 ±6.09)on the IED test. Between search error in the SWM test was positively correlation with YMRS scores ( r=0.38, P=0.039) in bipolar patients. Conclusion Both patient groups demonstrated a comparable profile of cognitive impairments during active periods of their condition. The cognitive impairment index may be a discreet cognitive endophenotype overlapping the disorders.