1.Liver cells and/or spleen cells injection induce islets transplantation resistance
Tianhua TANG ; Guosheng JIANG ; Fengqin JIANG ; Al ET
Chinese Journal of Immunology 1985;0(06):-
Objective:To observe the effect of liver cells and/or spleen cells injection induce islets transplantation resistance.Methods:New born male pigs and BALB/C mice were selected as donors and recipients respectively.The islets transplantation were performed in recipients just after tail vein injection with donor liver cells and/or spleen cells for 3 times.NK cells activity,antibodies forming function in vitro of B lymphocytes and T lymphocytes subsets measurement were used as immunological markers of transplantation resistance besides observation of the variation of blood glucose and xenograft living time(days).Results:The pre injection of donor liver cells,spleen cells or their mixture through mice tail vein was effective in preventing donor islets transplantation from rejection,which was demonstrated by the above immunological markers.And each kind of the transplantation could decrease the blood glucose of recipients and prolong the function possessed days of xenografts,especially for the more effective of mixture of donor liver cells and spleen cells as compared with the donor islets transplantation alone.Conclusion:Tail vein injection with donor liver cells and/or spleen cells could induce successfully islets transplantation resistance in mice.
2.Study of IFN? effect on HL-60 cells or ATRA resistant HL-60 cells
Guosheng JIANG ; Tianhua TANG ; Yukun ZHANG ; Al ET
Chinese Journal of Immunology 2000;0(11):-
Objective:To detect in vitro the effect of IFN? on HL 60 cells or ATRA resistant HL 60 cells Methods:MTT was used to measure the proliferation of HL 60 cells or ATRA resistant HL 60 cells The differentiation of HL 60 cells or ATRA resistant HL 60 cells was detected by NBT To detect the apoptosis of HL 60 cells or ATRA resistant HL 60 cells with Flow Cytometric Analysis Otherwise,the ATRA resistant HL 60 cells was induced by way of ATRA density gradually increasement Results:IFN? could inhibit the proliferation of HL 60 cells,especially accompanied with the ATRA IFN? not only induce the differentiation but also promote the differentiation of HL 60 cells or ATRA resistant HL 60 cells ATRA resistant HL 60 cells post treatment with IFN? were sensitive to ATRA Conclusion:Effect on differentiation of HL 60 cells was enhanced by IFN? IFN? also reverse the resistance of ATRA resistant HL 60 cells to ATRA
3.An Ultrasensitive Simultaneous Immunoassay Based on Arsenic and Mercury Ions Labeled SiO2@Au Nanoparticle Probes
Tianhua LI ; Ning GAN ; Dazhen WU ; Haijuan JIN ; Yuting CAO ; Qianli JIANG
Chinese Journal of Analytical Chemistry 2014;(6):817-823
An ultrasensitive immunoassay was developed based on As3+ and Hg2+ labeled SiO2 @ Au nanoparticles signal tags and hydride generation-atomic fluorescence spectrometry (HG-AFS) for the detection of carcinoembryonic antigen(CEA) and carbohydrate antigen 19-9 (CA 19-9) respectively. Firstly, amino SiO2@ Au NPs were synthesized for selective absorption of As3+ and Hg2+ ions respectively. Subsequently,the secondary antibody (Ab2) of CEA and CA 19-9 was respectively labeled on As3+ or Hg2+-SiO2 @ Au NPs to prepare the corresponding signal tags for CEA and CA 19-9. Based on the sandwich immunoassay scheme, the tags, two antigen and corresponding first antibodies were bio-conjugated on the bottom of 96-well plate at room temperature to form the immunocomplex. After it was dissolved in alkali solution, As3+ and Hg2+ ions were released in solution and detected by HG-AFS, which concentration was proportional with logarithms of CEA and CA 19-9. The reaction conditions were optimized and the tags were characterized. This assay was based on determination of the concentration of As3+ and Hg2+ for quantization of the corresponding CEA and CA 19-9 antigen. The assay showed a wide linear range from 0. 001 to 100. 0 μg / L for CEA and 0. 01-80 U/ mL for CA 19-9, and a lower detection limit of 0. 5 ng / L and 0. 005 U/ mL respectively. This proposed method was used in real serums samples, the results were consistence with that by ELISA. The immunoassay showed three orders of magnitude of sensitivity lower than that of ELISA, which provides a promising simultaneous immunoassay for the early diagnosis of cancer .
4.Synergetic effect of arsenic trioxide and Trichostatin A during inducing apoptosis of HL-60 cell and the molecular mechanisms
Weihua YANG ; Peie WEN ; Chao XIE ; Gaojuan QIAO ; Xia REN ; Haiquan REN ; Tianhua TANG ; Guosheng JIANG
Journal of International Oncology 2008;35(3):228-231
Objective To detect the synergetic effect and mechanism of arsenic trioxide(As2O3)and Trichostatin A(TSA)during inducing apoptosis of HL-60 cells.Methods MTT method was used to test the proliferation of HL-60 cells.Cell cycle and apoptosis were detected by FCM.Semi-quantitative RT-PCR was used to detect the mRNA expression of Bax and Bcl-2 in the cells treated by As2O3 and(or)TSA.Results As2O3 combined with TSA could inhibit proliferation and induce cell cycle arrest at G0 and G1.The percent of apoptosis induced by combination of As2O3 and TSA was obviously higher than that of either As2O3 or TSA.Bax gene expression was increased,while Bcl-2 gene expression was decreased,Bax/Bel-2 ratio was up-regulated.Conclusion Synergetic effect by As2O3 and TSA is remarkable in inducing apoptosis of HL-60 cells.Cell cycte arrest and Bax/Bcl-2 ratio play an important role in apoptosis of HL-60 cells induced by As2O3,TSA or their combination.
5.Effect of TGF-β1/SMAD signaling pathway on K562 cells growth inhibition caused by HMBA
Enyu SU ; Peie WEN ; Xia REN ; Xiaobai SUN ; Henglan ZHANG ; Tianhua TANG ; Haiquan REN ; Guosheng JIANG
Journal of International Oncology 2010;37(4):312-315
Objective To investigate the effect of TGF-β1/SMAD signaling pathway on K562 cells growth inhibition caused by HMBA. Methods After establishing the in vitro differentiation model with HMBA on K562 cells, the MTT assay was used to detect the proliferation of K562 cells, the cell cycle profile was detected by flow cytometry, and the mRNA expression of TGF-β1, SMAD3, SMAD4 and EVI1 was measured by RT-PCR assay. Results HMBA could inhibit the proliferation and promote the differentiation of K562 cells obviously, which was time and concentration-dependent, and the 72 h corresponding IC50, was about 2 mmol/L. Within 72 h, flow cytometry assay indicated that the ration of G0-G1 phase cells was up-regulated, and the results of RT-PCR showed that relative mRNA expression of TGF-β1, SMAD3 and SMAD4 at mRNA level was increased gradually while that of EVI1 was decreased gradually. Conclusion HMBA can inhibit K562 cells proliferation through TGF-β1/SMAD signaling pathway.
6.Screening of proteins interacting with RSK4 in breast cancer by affinity purification and liquid chromatography/mass spectrometry
Riqiang LIU ; Tianhua LIU ; Qinle ZHANG ; Kai JIANG ; Kun GUO ; Shu ZHANG ; Yinkun LIU ; Huawei YANG
China Oncology 2016;26(7):581-588
Background and purpose:As a tumor suppressor gene, ribosomal S6 kinase 4 (RSK4) plays important roles in inhibiting cell proliferation, migration and inducing cell apoptosis. However, the proteins interacting with RSK4 are still unknown. This study aimed to screen proteins interacting with RSK4 in breast cancer cell line MDA-MB-231 by lfag-tag affnity puriifcation and LC-MS/MS (liquid chromatography/mass spectrometry).Methods:The pcDNA3.1/EGFP-RSK4-Flag eukaryotic expression vector was constructed by inserting full lengthRSK4 gene into vector pcDNA3.1/EGFP-Flag. And then the recombinant plasmids were transferred into MDA-MB-231 cells. Real-timelfuorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot were used to detect the expression of RSK4 in MDA-MB-231 cells. Affnity puriifcation and LC-MS/MS were applied to screen proteins interacting with RSK4, and the related action mechanism of RSK4 with its interacted proteins was detected based on bioinformatics gene ontology (GO) and ingenuity pathway analysis (IPA).Results:Twenty-four proteins, such as serine/threonine-pro-tein kinase 38 (STK38)/serine/threonine-protein kinase 38-like (STK38L), MOB kinase activator 2 (MOB2) and protein arginineN-methyltransferase 5 (PRMT5), were successfully identiifed by Flag-tag affnity puriifcation followed by LC-MS/MS analysis, which probably interacted with RSK4. Bioinformatics analysis of the identiifed proteins suggested the proteins interacting with RSK4 were involved in diverse biological pathways, such as apoptosis and cell migration. Conclusion:According to bioinformatics results of proteins interacting with RSK4 identiifed by affnity puriifcation and LC-MS/MS, biological networks of RSK4 are involved in apoptosis and migration in breast cancer cells.
7.Analgesic effects of new triazole compounds Ⅱ_3 and their effects on the activity of cycloxygenase
Yongna ZHANG ; Tianhua YAN ; Qiujuan WANG ; Bo JIANG ; Jinyi XU ; Xiaoming WU
Journal of China Pharmaceutical University 2009;40(6):539-543
Aim: To investigate the analgesic effect of the new triazole compounds Ⅱ_3 and effects on cycloxygen-ase-1(COX-1) as well as cycloxygenase-2( COX-2). Methods: The hot plate and the stretching settings in mice were utilized to study the effects of compounds Ⅱ_3 on acute pain. Radioimmunologic kits were used to assay the contents of PGE_2 in macrophage and 6-keto-PGF_(1α) in endodermis, which represents the activities of COX-2 and COX-1, respectively. Results: CompoundsⅡ_3( 15,30,60 mg/kg) prolonged the pain liminal value and the writ-hing response time in the initial appearance, and reduced the frequency of the writhing response in 15 min after exposure of the mice to glacial acetic acid( P < 0. 05, P < 0. 01). CompoundsⅡ_3, at the concentrations of 1×10 ~(-5),1×10 ~(-6), and 1×10 ~(-7) mol/L, markedly inhibited the production of PGE_2 in macrophage, and also impeded the activity of COX-2 at 1×10 ~(-6) mol/L But the inhibition of 6-keto-PGFla in endodermis using the same settings of compounds Ⅱ_3 was found to be limited. Conclusion: CompoundsⅡ_3 has analgesic effects on the acute pain and selective inhibition on COX-2.
8.Study on mechanism of action of promoting blood circulation and dispersing phlegm method in improving insulin resistance of type 2 diabetes rats
Jianhua FENG ; Guosheng JIANG ; Yunsheng XU ; Baorong GUO ; Tianhua TANG ; Peie WEN ; Baohua DONG
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(08):-
0.05).Compared with control group,the GK activity of liver cell,the expression of PEPCK and the expression of GLUT4 in model group decreased signifi cantly(P
9.CT Diagnosis of Split Cord Malformation in Children
Bo LIU ; Zengjun ZHANG ; Tianhua CHEN ; Jufen JIANG ; Yuqin CAO ; Mingxia HUANG
Journal of Practical Radiology 1991;0(03):-
Objective To study the appearances of split cord malformation(SCM)and evaluate the diagnostic value of CT for SCM.Methods Clinical and CT data of 48 cases with SCM were analyzed retrospectively ,21 were males and 27 were females,ranged from 1 day to 8 years with a mean of 11.6 months. All cases evaluated by plain CT with coronal and sagittal reconstructions.Results Type I accounted 75%, consisted of two hemicords, each contained in its dural tube and separated by a rigid median septum .TypeⅡaccounted 25%, consisted two hemicords contained in a single dural sac separated by a non-rigid, fibrous median septum. Associated abnormalities: tethered cord syndrome(n=38), syringomyelia(n=9), intradural lipomas(n=10), meningocele(n=18).Conclusion CT can clearly demostrate the position, the septum and the shape of the SCM, as well as associated abnormalities.
10.Reversal of multidrug resistance of K562/ADM cells in vitro by the monomer of traditional Chinese medicine extract and investigation of its molecular mechanism
Gaojuan QIAO ; Guihai LI ; Weihua YANG ; Peie WEN ; Xia REN ; Hua FAN ; Tianhua TANG ; Haiquan REN ; Guosheng JIANG
Journal of International Oncology 2009;36(6):474-476
Objective To investigate the reversal effect of the monomer of traditional Chinese medicine on muhidrug resistance(MDR) and its possible mechanism in K562/ADM cell line in vitro. Methods With different concentrations of baicalin, geniposide administered to K562/ADM cells, the proliferation of K562/ ADM cells was detected by the MTY assay. Expression of mdr-1 mRNA, Topo Ⅱ mRNA was measured by semi-quantitive RT-PCR. Results Thatbaicalin and geniposide could increase the sensitivity of K562/ADM cells to adriamycin, multiples of reversion were 1.95 times and 1.46 times. The proliferation of K562/ADM cells was in-hibited obviously by baicalin and geniposide, the level of mdr-1 mRNA expression was down-regulated and the Topo Ⅱ mRNA was up-regulated(P<0.01 ). Conclusion Baicalin and geniposide may reverse the multi-drag-resistance of K562/ADM cells, which was related to the down-regulation of mdr-1 expression and up-reg-ulation of Topo Ⅱ beta expression.