2.Comparison of inhibitory effects between artemisinin and artemisunate on proliferation of MCF-7 cells in vitro
Fang LIN ; Zhiyu QIAN ; Hongwei XUE ; Jian DING ; Liping LIN
Chinese Traditional and Herbal Drugs 1994;0(04):-
Object To study the effects of artemisinin and its analogue artemisunate on the proliferation of human breast cancer MCF-7 cell line, as well as their mechanism comparatively. Methods The inhibition of cell proliferation was determined by SRB method. Cell cycle was determined by flow cytometry (FCM) analysis. Apoptosis was confirmed by sub-G 1 cells content and DAPI method. Results The cell cycle of MCF-7 was changed greatly when treated 24 h with either 10 ?mol/L artemisinin or 1 ?mol/L artemisunate, the distribution of MCF-7 cells among S phase was reduced greatly, while inereased during G 0+G 1. However, artemisinin had weaker effect on the proliferation of MCF-7 cell, while artemisunate effectively inhibited the proliferation of MCF-7, the IC 50 was 0.31 ?mol/L. Apoptosis induced by 1 ?mol/L artemisunate was stronger than that by 10 ?mol/L artemisinin, too. Conclusion The inhibitory effect of artemisunate on the proliferation of tumor cell is stronger than that of artemisinin in vitro.
3.Effect of Notch1 overexpression on proliferation of cancer cell lines
hai, YU ; sheng-fang, GE ; jian, LU ; guan-xiang, QIAN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(03):-
Objective To upregulate Notch signaling in cancer cells by overexpression of active part of Notch1 and to examine the proliferation of the cells. Methods Four cancer cell lines were infected with retrovirus recombined with sequence encoding active part of Notch1.CBF-1 reporter plasmid was used to detect Notch signaling and proliferation assay was carried out by MTS method.Cell cycle analysis was synchronously conducted. Results The overexpression of the active part of Notch1 induced upregulation of Notch signaling,led to growth inhibition in Hela and HepG2 cell lines and growth boost in BGC-823 cell lines,while had no effect on Chang cell lines. Conclusion The upregulation of Notch signaling can exert various effects on different cancer cell lines which is critical to the gene therapy for cancers.
4.Survioin promoter activity in tumor cell lines
rang, XU ; sheng-fang, GE ; jian, LU ; guan-xiang, QIAN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(08):-
Objective To study the activity of the survivin gene promoter in several tumor cell lines and evaluate the possible application of this promoter in tumor gene therapy. Methods ①The expressions of survivin gene in A549,MDA-MB231 and HepG2 cell lines were detected by RT-PCR and Western blotting.②Tumor cells(A549,MDA-MB231,HepG2) were transiently transfected by reporter plasmids containing different length of survivin promoter using lipofectamine.And 48 h later,the level of reporter gene expression was analyzed.Results There were different levels of survivin expression in A549,MDA-MB231 and HepG2 cell lines.Transient transfection assay approved that pLuc-surP-987,pLuc-surP-596,pLuc-surP-269 and pLuc-surP-158 showed high activity and 269 bp survivin promoter demonstrated the highest activity. Conclusion In transcriptional level,survivin promoter can activate the reporter gene in several tumor cell lines.It is a potential candidate promoter in tumor gene therapy.
5.The effects of endotoxin on the Th1/Th2 cells and immune modulation of Astragalus membranaceus.
Jian-hua LIU ; Ying-qian ZHANG ; Bo HUANG ; Qian FANG ; Ximin HUO ; Haofu HU
Chinese Journal of Pediatrics 2003;41(8):613-614
Animals
;
Astragalus membranaceus
;
Endotoxemia
;
drug therapy
;
metabolism
;
Interferon-gamma
;
analysis
;
Interleukin-4
;
analysis
;
Male
;
Mice
;
Phytotherapy
;
Plant Extracts
;
therapeutic use
;
Th1 Cells
;
drug effects
;
metabolism
;
Th2 Cells
;
drug effects
;
metabolism
6.Etiology and clinical features of hepatic dysfunction in patients after allogeneic hematopoietic stem cell transplantation
Qian WANG ; Yuewen FU ; Yanli ZHANG ; Baijun FANG ; Jian ZHOU ; Xudong WEI ; Yongping SONG
Journal of Leukemia & Lymphoma 2012;21(8):477-480
Objective To summarize and evaluate the incidence,etiology,diagnostic and therapeutic method of hepatic dysfunction after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods 83 blood disease patients who undergoing allo-HSCT from 2006 to 2010 in the affiliated cancer hospital of Zhengzhou university.Among those who suffered from Ⅱ-Ⅳ grade hepatic dysfunction,the incidence,the ratio of different causes,clinical feature and diagnostic method were evaluated.The difference of causes of hepatic dysfunction in different period,the therapeutic method and curative effect were also analysed.Results Among 83 patients undergoing allo-HSCT,45 patients suffered from Ⅱ-Ⅳ grade hepatic dysfunction,the ratio was 54.2 %.For etiology,7 were preconditioning,9 were cyclosporine (CsA),2 were hepatic venoocclusive disease (HVOD),24 were hepatic graft versus host disease (GVHD),2 was hepatic B virus (HBV)reactivation,1 was mutiple organ failure.20 cases (44.4 %) occurred in one month after allo-HSCT with the main etiology of drug hepatotoxicity.13 cases (28.9 %) occurred from one month to 100 days after allo-HSCT,while 12 cases (26.7 %) occurred from 101 days to one year with the main etiology of both hepatic GVHD.27 cases were cured and 10 were improved after treatment.2 cases were not cured and 6 cases died from relapse of the primary disease,or else from the complication of allo-HSCT.Conclusion Hepatic dysfunction is an common complication after allo-HSCT,drug hepatotoxicity and hepatic GVHD are the major causes.The relativity between hepatic dysfunction and period after allo-HSCT is a important reference for diagnosis.It will produce desired result to choose proper therapeutic method based on etiology.
7.Ag85B regulates myeloid dendritic cell maturation and suppresses expres-sion of TSLPR and OX40L mediated by TSLP in vitro
Jiang QIAN ; Jian WU ; Hong AN ; Xiangfeng FANG ; Dongfeng LI ; Shifang YANG ; Jinxiu MENG ; Xinglin GAO
Chinese Journal of Pathophysiology 2015;(9):1680-1687
AIM:To investigate the maturation of mice immature myeloid dendritic cells (mDCs) induced by antigen(Ag)85B of mycobacterium tuberculosis, and the expression of TSLPR and OX40L mediated by TSLP in vitro. METHODS:Recombinant mouse GM-CSF ( rmGM-CSF) and rmIL-4 were used to induce bone marrow precursor cells of C57BL/6 mice to differentiate into immature mDCs in vitro.mDCs were identified followed by purification using CD 11c binding magnetic beads .The morphological characteristic of mDCs was observed under inverted phase-contrast microscope and scanning electron microscope .The surface phenotypes of mDCs were determined by flow cytometry .To obtain the opti-mal concentrations of Ag85B and TSLP, immature mDCs were cultured with different concentrations of Ag 85B or TSLP at 0 (control group), 50, 100 and 200 μg/L for 24 h, and the expression of cell surface molecules CD 80, CD86, TSLPR and OX40L was detected by flow cytometry.In addition, the expression of TSLPR and OX40L in Ag85B and TSLP-co-stimula-ted mDCs was determined by flow cytometry .RESULTS:After 7 d of culture in vitro, the cells showed irregular dendritic protrusions under the inverted-phase contrast microscope , and had wrinkles and dendritic splits under scanning electron mi-croscope , conformed to the morphological characteristics of immature mDCs .The mDCs cells expressed higher level of spe-cific marker CD11c, lower level of co-stimulatory molecules CD80 and CD86, which conformed to the phenotype of imma-ture mDCs.The CD80 +and CD86 +cell ratios of mDCs displayed significant increases in 50, 100 and 200μg/L Ag85B or TSLP groups compared with control group (P<0.05).The ratios of TSLPR +and OX40L+cells did not differ among dif-ferent concentrations of Ag 85B groups.The ratios of TSLPR +and OX40L+cells were significantly increased in 100 μg/L and 200μg/L TSLP groups compared with control group and 50μg/L TSLP group (P<0.05).Under the circumstance of optimal Ag85B or TSLP treatment concentration at 200 μg/L, there was significantly decreased in TSLPR and OX 40L cell ratio of mDCs in Ag85B group or Ag85B combined with TSLP group when compared with TSLP group (P<0.05), and no significant difference among Ag85B group, Ag85B combined with TSLP group and control group was observed .CONCLU-SION: Ag85B enhances mDCs maturation by up-regulating the expression of co-stimulatory molecules CD80 and CD86, and inhibit the expression of pro-inflammatory specific molecules TSLPR and OX40L on TSLP-activated mDCs, indicating that Ag85B modifies the development of asthmatic airway inflammation through the pathway of TSLP -activated mDCs.
8.Molecular characteristics of pyrazinamide resistance in Mycobacterium tuberculosis strains
Zhenhua LUO ; Xueqin QIAN ; Qiwen FAN ; Huanying FANG ; Jian GUO ; Wenjuan WU
Chinese Journal of Microbiology and Immunology 2015;(9):660-665
Objective To investigate the correlation between the mutation of pncA gene and the susceptibility to pyrazinamide ( PZA) in Mycobacterium tuberculosis complex ( MTBC) strains and to analyze the mutation of panD and rpsA genes in wild type isolates without pncA gene mutation.Methods The sus-ceptibilities of 108 MTBC strains to first-line drugs including PZA were detected by using the MGIT 960 TB system.PCR was performed to amplify the 16S rDNA and pncA, panD and rpsA genes.The PCR products were analyzed by DNA sequencing analysis .Results Among the 78 multidrug-resistant MTBC strains , 47 isolates (60%) were resistant to PZA.Four out of 30 (13%) strains that were sensitive to ethambutol , iso-niazid, rifampicin and streptomycin (EIRS) were resistant to PZA.The drug-resistant MTBC strains showed higher resistance rate to PZA than that of the EIRS sensitive strains .There were 49 ( 96%) PZA-resistant isolates and 4 (7%) PZA-sensitive isolates occurred pncA gene mutation.Most of the pncA gene mutations in the genomes of PZA-resistant strains were base substitution mutation , especially the His57Asp substitu-tion.The pncA gene mutations centralized in the regions of 160-169, 203-289, 309-396 and 413-467.Seven novel mutation sites of pncA gene were observed including T175C, C188A, G insertion at 68, AGC insertion at 235, C insertion at 339, CC insertion at 392 and GT deletion at 395.The mutation sites founded in the genomes of PZA-sensitive strains were different from those of the PZA-resistant strains .No mutation of the pncA gene and the upstream regulatory sequence was found in two PZA-resistant strains , NJ44 and NJ108 . The sequence analysis of panD and rpsA gene showed that the NJ 108 strain had panD gene mutation at G419A, but no mutation was detected in the NJ 44 strain.Conclusion The multidrug-resistant MTBC strains showed higher resistance rate to PZA .The pncA gene mutation was common in PZA-resistant MTB strains and the panD gene mutation was also worthy of attention .
9.The protective effect of aminophylline on myocardium during cardiopulmonary bypass
Jian-Fang QIAN ; Ai-Qiang DONG ; Wan-Jun LUO ; Guan-Yu JIANG ;
Chinese Journal of Emergency Medicine 2006;0(12):-
Objective To evaluate the protective effect of aminophylline on myocardium in the patients undergoing prothetic valve replacement operation of heart.Mothods Thirty patients undergoing prothetic valve replacement operation of heart were randomized to be treated either with aminophylline(n=15)or without aminophylline treatment(n=15). Aminophylline(5mg/kg)was injected intravenously at 15 minutes after induction of anesthesia.Cardiac troponin I(cTnI), cyclic adenosine monophosphate(cAMP),myeloperoxidase(MPO),ratio of aortic blood neutrophil count to coronary vein sinus blood neutrophil count,hemodynamics,time of aortic cress-clamping and other clinical data were recorded during the operation.Results There were no differences between the two groups in the major perioperative variables.Plasm cTnI concentration in both groups increased after off-clamping than that before CPB,however,it was lower in aminophylline group than that in control group.Concentration of cAMP in both groups after off-clamping was lower than that before CPB, however cAMP concentration in aminophylline group after off-clamping was higher than that in control group.Myocardial MPO activity and neutrophil count ratio after aortic off-clamping in aminophylline group was significantly lower than that in control group.Conclusion These results suggest that aminophylline is helpful to unprotection of myocardial and decreases the sequestration of neutrophil in myocardium.The mechanism of the protection may be related to the cAMP increased in myocardium.
10.Early application of non-invasive ventilation in treating elderly patients with acute left ventricular failure and respiratory failure
Bao-Min FANG ; Tie-Ying SUN ; Hui-Xing KE ; Jin SHEN ; Yi-Jian QIAN ;
Chinese Journal of Emergency Medicine 2006;0(06):-
Objective To study effects of early non-invasive ventilation (NIV) application in treating elderly patients with acute left ventricular failure induced respiratory failure. Method Totally 32 elderly patients with acute left ventrieular failure induced respiratory failure, admitted from August 1997 to February 2007, received NIV treatment, and were retrospectively studied. There were 22 male and 10 female, aged (81.5?8.6) yearsdd. The changes of rahs, respiration rate, heart rate, arterial blood gas, cardiac function before and after NIV application were compared. According to the application time of NIV, 32 patients were divided into two groups: group A (early NIV application group, n=17) and group B (non-early NIV application group, n= 15). The time to improve the symptoms, the application time of NIV, cure rates, tracheal intubation rates and mortality were compared between the two groups. Results Thirty of the 32 patients survived, cardiac function was improved from New York classⅣtoⅠ~Ⅱ, respiratory rate, heart rate and blood pressure significantly decreased, PaO_2 and SaO_2 significantly increased and PaCO_2 significantly decreased. The tracheal intubation was performed in 4 patients. The time needed to improve the symptoms and the application time of NIV were significantly different between group A and group B (P