3.Eexpression of Foxp3 and CD4~+CD_(25)~+ Regulation T Cells in Children with Acute Idiopathic Thrombocytopenic Purpura
xue-wen, LV ; fang, LIU ; chang-lin, WU
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To investigate the amount of CD4+CD_(25)~+ regulation T cells(Tr cells) and the expression of Foxp3 in peripheral blood in children with acute idiopathic thrombocytopenic purpura(AITP),and analyze the relationship between CD4+CD_(25)~+ Tr cells and the immunopathogenesis of AITP.Methods CD4+CD_(25)~+ Tr cells were detected by flow cytometry in peripheral blood from AITP children and health children.Besides,related cytokine in peripheral blood were assigned by enzyme linked immunosorbent assay(ELISA).The expression of Foxp3 were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Results CD4+CD_(25)~+ Tr cells decreased obviously in peripheral blood in children compared with that of control group [(2.83?1.05)% vs (5.07?0.59)%,P
4.Expression of glutathione S-transferase mRNA in peripheral blood of the population in coal-burning fluorosis area
Chang-xue, WU ; Ke-ren, SHAN ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2010;29(2):121-123
Objective To investigate expression of glutathione S-transferase(GST) mRNA in peripheral blood of the population in coal-burning fluorosis area and to evaluate the effect of comprehensive control intervention. Methods Fifty samples of peripheral blood from patients in the coal-buring fluorosis area in Bijie county of Guizhou province were selected as fluorasis group and 50 samples of peripheral blood from patients in area with comprehensive management were selected as intervention group, respectively. Fifty samples from non-endemic fluorosis area were selected as the control group. Total RNA from blood was extracted and purified by the Trizol- Phenol-Chloroform one-step method. Expression of GST mRNA was detected by using SYBR Green I real-time fluorescence quantitative PCR. Results The data of GST mRNA in fluorosis group, intervention group and control group was 38.28±27.22,70.56±37.23 and 103.46 ± 46.62, respectively. There was a significant difference between the groups(F = 3.75, P < 0.05). Decreased expression of GST mRNA in fluorosis group and intervention group as compare to control was detected(all P < 0.05), and the expression of GST mRNA in intervention group was higher than that in fluorosis group(P < 0.05). Conclusion Coal-burning fluorosis possibly led to the decreased expression of GST mRNA in peripheral blood, and comprehensive control maybe prevent the decreased expression of GST in mRNA level.
5.Significance of Protein Kinase C Activity Detection in Children with Chronic Idiopathic Thrombocytopenic Purpura
chang-lin, WU ; jian-cheng, XUE ; xue-min, ZHOU ; fang, LIU
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To investigate the changes of protein kinase C(PKC)activity in peripheral blood T lymphocytes in children with chronic idiopathic thrombocytopenic purpura(CITP)and the relationship between PKC activity,T lymphocytes activation and thrombocytes decrease.Methods Collecting sterile peripheral blood from CITP children(n=30)and healthy children(n=30),T lymphocytes were isolated and purified by the T cell segregation enrichment column,the PKC activity was detected by non-radioactive assay.Soluble interleukin-2 receptor(sIL-2R),which was T cell activated marker,was determined by enzyme-linked immunoabsorbent assay(ELISA),platelet was counted by cell counting meter.Results Compared with healthy children,PKC activity was significantly enhanced in CITP children[(0.94?0.23)mmol/(min?L)vs(0.50?0.17)mmol/(min?L),t= 8.42 P
6.Effects of α3 neuronal nicotinic acetylcholine receptor on cell apoptosis and p38 MAPK signal transduction pathway in SH-SY5Y cells.
Xue-ling ZHANG ; Xiao-lan QI ; Jia-mou REN ; Chang-xue WU ; Zhi-zhong GUAN
Chinese Journal of Pathology 2013;42(2):116-120
OBJECTIVETo investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD).
METHODSThe levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again.
RESULTSCompared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former.
CONCLUSIONThe siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.
Alzheimer Disease ; etiology ; Amyloid beta-Peptides ; metabolism ; Apoptosis ; Cell Line, Tumor ; Gene Silencing ; Humans ; Neuroblastoma ; metabolism ; pathology ; Peptide Fragments ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Nicotinic ; genetics ; metabolism ; Signal Transduction ; Transfection ; bcl-2-Associated X Protein ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism
8.Research on a tool for organizational culture assessment of public general hospitals in China
Ping ZHOU ; Jile CHANG ; Jinxing HUANG ; Lan LI ; Zhiliu TANG ; Jiayan HUANG ; Rong WU ; Di XUE
Chinese Journal of Hospital Administration 2011;27(6):433-436
Objective A scientific evaluation of hospital culture with the Dimension organizational culture model, in view of features of China's general public hospitals.Methods Based on Denison model, according to the characteristics of the public general hospitals in China, the authors developed a tool for organizational culture assessment (TOCA) by using the survey data from 87 hospitals in three provinces from the East, Central, and West areas in China.Results This tool, an evaluation scale, comprises the four cultural characteristics of direction, consistency, participation, and adaptability, as well as 13 cultural dimensions of social responsibility and competitive consciousness. The tool is tested as having good internal reliability and validity.Conclusion The TOCA provides hospital administrators with a tool for hospital culture evaluation, diagnosis and improvement.
9.Abnormal NT5C2 gene expression in acute lymphoblastic leukemia
Xia WU ; Lei HAO ; Bing XUE ; Dandan HAN ; Renliang CHANG ; Fang DENG
Chinese Journal of Laboratory Medicine 2016;39(7):501-505
Objective To investigate the expression and significance of mRNAand exon mutationof NT5C2 gene in acute lymphoblastic leukemia (ALL) bone marrow.Methods Case control study design was used in this study.Bone marrow samples were collected from ALL patients in Anhui Provincial Cancer Hospital in recent 4 years.The patientswere divided into the initial diagnosis group , the complete remission group and the recurrence group.And they could specifically be divided into 36 patients initially diagnosed, 36 patients who achievedcomplete remission and 16 patients who relapsed with children B -ALL,15 patients initially diagnosed,15 patients who achievedcomplete remission and 9 patients who relapsed with children T -ALL, 18 patients initially diagnosed,18 patients who achievedcomplete remission and 12 patients who relapsed with adult B-ALL, and 11 patients initially diagnosed,11 patients who achievedcomplete remission and 6 patients who relapsed with adult B -ALL.The initial diagnosis,complete remission and recurrence samples were matched.8 children and 8 adults without hematologic malignanciewere used as controls .Real-time PCR was performed to detect the level of NT5C2 mRNAin ALL patients.The exons of NT5C2 gene were cloned and sequenced for the common mutations in all cases .The results of NT5C2 mRNA levels in different groups were performed using non -parametric test by SPSS16.0 analytics software, and then non-parametric test together with correlation analysis was analyzed between NT 5C2 mRNA levels of different initial diagnosis groups and gender, age, leukocyte level and risk classification .Results (1)The expression of NT5C2 mRNA levels of recurrence group were higher than that of initial diagnosis group ,complete remission group and controls in children and adult B -ALL respectively(P <0.01).(2)NT5C2 mRNA expression in children and adult T-ALL showed no difference in initial diagnosis ,complete remission, recurrence group and controls (P >0.05).(3)NT5C2 mRNA expression of initial diagnosis group in children and adult B -ALL and T-ALL was not correlated with risk classification (P >0.05).(4)A newheterozygousmutation p.P414A of NT5C2 was discovered in a recurrencesample.Conclusions (1) High expression ofNT5C2 mRNA is associated with recurrence inchildren and adult B-ALL, and it may be an indicator of monitoring recurrence .(2)The incidence of exons mutation of NT5C2 gene in ALL is low in China.
10.Effect of yuejuganmaidazaotang in hippocampus in postpartum depression model mice on AKT/mTOR signaling pathway
Baomei XIA ; Chang CHEN ; Hailou ZHANG ; Wenda XUE ; Ruyan WU ; Li REN ; Weiwei TAO ; Gang CHEN
Chinese Pharmacological Bulletin 2016;(1):119-122
Aim Using chronic pre-pregnancy stress to establish a postpartum depression animal model, given a single YG,and acute ketamine was served as control, to explore the pathology of PPD and the anti-depressive mechanism of the YG on the PPD model on AKT/mTOR signaling pathway. Methods Thirty-two fe-male Balb / c were randomly assigned to two groups, the control group ( Control, Con) and the pre-pregnancy stressed group(Model,Mod) , which was subjected to 3 weeks chronic restraint stress. After the last stressor, the pre-pregnancy stressed group was housed with a male. After about 4 weeks later, the mice gave birth to pups. Then at 3 weeks postpartum, we tested the ma-ternal tail suspension test ( TST). Both YG and Ket-amine was single administered 24 hours before behavior test, with single saline for control group and PPD mod-el group. After TST,the mouse hippocampus were ex-tracted to detect the expression of AKT and mTOR. Results After 3 weeks postpartum, the model mice showed depression-like behaviors. Immobility in TST was significantly increased in vehicle groups(P <0. 01). Acute YG improved performance in the TST (P< 0. 01), which was similar to ketamine. And the PPD model mice group showed decreased phosphorylation of AKT and mTOR (P < 0. 01,P < 0. 01), compared to control group. A single dose of YG or ketamine normal-ized AKT/ mTOR signaling in the PPD model mice(P< 0. 01,P < 0. 01),( P < 0. 01,P < 0. 01). Conclu-sions Chronic pre-pregnancy stress can induce dams into postpartum depression and its mechanism maybe associated with down-regulating AKT/ mTOR signa-ling. Acute YG exerts fast antidepressant effect on this PPD model similar to ketamine, and its mechanism may be related to up-regulating AKT/ mTOR signaling in the hippocampus.