1.Effect of NF-κB inhibitor pyrrolidine dithiocarbonate on the proliferation and apoptosis in K562 cells
Chun ZHANG ; Guohui CUI ; Fang LIU
Chinese Journal of Pathophysiology 2009;25(12):2349-2352
AIM: To investigate the effect of pyrrolidine dithiocarbamate (PDTC), a specific inhibitor of NF-κB on the proliferation and apoptosis of K562 cells and to explore the anti-tumor mechanism of PDTC.METHODS: Trans AM~(TM) NF-κB p65 kit was used to detect the activity of p65 in K562 cells treated by PDTC. The effect of PDTC on the proliferation of K562 cells was measured by WST-1 method. DNA damage was detected by single cell gel electrophoresis (comet assay). The procaspase-3 and activated protein level of caspase-3 were detected by Western blotting.RESULTS: The activity of p65 in K562 cells was inhibited after treated by PDTC (P<0.01). Simultaneously the cell proliferation was significantly inhibited in a dose-and time-dependent manner (P<0.01). The degree of DNA damage in K562 cells treated with PDTC at concentrations of 25 μmol/L, 50 μmol/L or 100 μmol/L was more severe than that in control. The rates of comet cells in the PDTC-treated groups (43.50%, 84.00%, 95.63%) were significantly higher than those in control (9.75%, P<0.01), and it was also dose-dependent. The expression of procaspase-3 and activated caspase-3 protein were detected in the cytoplasm of the K562 cells treated by PDTC by Western blotting.CONCLUSION: NF-κB plays an important role in regulating cell proliferation and apoptosis in K562 cells. PDTC inhibits NF-κB activity and elevates the expression of caspase-3, which is related to increase in cell apoptosis.
2.Effect of NF-?B inhibitor pyrrolidine dithiocarbonate on the proliferation and apoptosis in K562 cells
Chun ZHANG ; Guohui CUI ; Fang LIU
Chinese Journal of Pathophysiology 2000;0(12):-
AIM:To investigate the effect of pyrrolidine dithiocarbamate (PDTC),a specific inhibitor of NF-?B on the proliferation and apoptosis of K562 cells and to explore the anti-tumor mechanism of PDTC.METHODS:Trans AMTM NF-?B p65 kit was used to detect the activity of p65 in K562 cells treated by PDTC. The effect of PDTC on the proliferation of K562 cells was measured by WST-1 method. DNA damage was detected by single cell gel electrophoresis (comet assay). The procaspase-3 and activated protein level of caspase-3 were detected by Western blotting.RESULTS:The activity of p65 in K562 cells was inhibited after treated by PDTC (P
3.ELECTRON MICROSCOPIC OBSERVATIONS ON SYNAPSES IN THE HYPOTHALAMIC VENTROMEDIAL NUCLEUS OF THE RAT
Mingxuan ZHANG ; Chun MA ; Veinan FANG
Acta Anatomica Sinica 1957;0(04):-
The types of synapses and some morphological features of the synaptic submicroscopic structures of rat hypothalamic ventromedial nucleus (HVM) have been described in this study. A total of 1 005 synapses is observed. The percentage of axo-dendritic synapse is 96.6%. It covers the majority of the total number of synapses. The percentage of axo-somatic synapse is 2.8% and axo-axonic synapse is 0.5% of the total number of the synapses. The crest synapse which is never reported in the HVM was found to be 0.1%. Its postsynaptic elements come from clubbed finger-like dendritic crest which is characterized by prominent postsynaptic membrane thickening and subsynaptic dense bodies. Two presynaptic terminals contact the wall of the crest process side by side and contain spheric vesicles. In the axo-dendritic and axo-somatic synapses, the parallel synapse (including synaptic complex), the tangent-like synapse and serial-like synapse have been observed.
4.An open-label trial on the safety of recombinant human tumor necrosis factor-Fc treatment in Chinese rheumatic diseases patients
Fang DU ; Chun-de BAO ; Feng-chun ZHANG
Chinese Journal of Rheumatology 2011;15(12):850-854
Objective To compare the safety of recombinant human tumor necrosis factor-Fc (rhTNFR:Fc) and other DMARDs,in patients with rheumatoid arthritis (RA),ankylosing spondylitis (AS),juvenile idiopathetic arthritis (JIA) or psoriatic arthritis (PsA).Methods Patients who received rhTNFR:Fc 25 mg twice weekly from May 2006 to March 2009 were involved in this open-lable study.Safety assessments were carried out at regular intervals.Results Of the 2014 patients enrolled in the open-label trial,1388,421 and 232 were RA,AS or other diseases,such as JIA and P.sA respectively.Frequent adverse events included injection-site reactions (2.67%),rash (1.87%) and hyperamino transferase (1.80%) in RA patients.Similarly,injection-site reactions (5.23%),hyperaminotransferase (2.38%) and rash (0.71%) were frequent in AS patients.Upper respiratory infection was most frequent among infectious adverse events.There were no reports of patients with serious adverse events,dead case,TB infection and malignancies.Conclusion rhTNFR:Fc has shown a favourable safety profile in Chinese rheumatic disease patients.
5.The relationship between serum tumor markers and the severity of silicosis
Shencun FANG ; Haitao ZHANG ; Bin XU ; Chun WANG ; Yingming ZHANG
Chinese Journal of Laboratory Medicine 2017;40(7):515-519
Objective To investigate the relationship between serum tumor markers and the severity of silicosis.Methods Retrospective study.Total of 160 patients with silicosis were included in the study, and 160 healthy volunteers were recruited as the control group.Tumor marker levels in both bronchoalveolar lavage fluid(BALF)and serum were detected by the immunochemiluminecence methods.The pulmonary function parameters, blood gas analysis and lactate dehydrogenase(LDH)were also analyzed.Lung tissue obtained by a patient with silicosis was stained by neuron specific enolase(NSE), carbohydrate antigen125(CA125) and carbohydrate antigen19-9(CA19-9).Results Serum NSE, CA125 and CA19-9 levels were significantly higher in cases than those in controls[(34.47±13.30)μg/L vs(10.24±7.20)μg/L,t=20.27, P<0.000 1;(33.96±17.80)kU/L vs(12.23±15.30)kU/L, t=11.71, P<0.000 1;(4.68±5.67)kU/L vs(2.78±3.45)kU/L,t=3.67,P<0.002].Significant negative correlations were found between values of tumor markers(CA125 and CA19-9) and spirometric parameters,such as forced expiratory volume in one second %(FVE1%), forced expiratory volume in one second/forced vital capacity(FVE1/FVC), carbon monoxide diffusion capacity (Dlco) and total lung capacity(TLC) (r=-0.423,P=0.001;r=-0.323,P=0.011;r=-0.479,P=0.001;r=-0.285,P=0.043) and (r=-0.324,P=0.022;r=-0.256,P=0.023;r=-0.354,P=0.013;r=-0.356,P=0.012).Significant positive correlations were also observed between values of these tumor markers and LDH(r=0.378,P=0.001 and r=0.347,P=0.21).Significant negative correlations were found between NSE and Dlco and TLC(r=-0.374,P=0.004 and r=-0.368,P=0.002).Significant positive correlations were also observed between NSE and LDH(r=0.404,P=0.001).The NSE and CA19-9 levels in BALF were significantly higher than those in serum[(39.32±29.30)μg/L vs(25.7±12.12)μg/L,t=2.15,P=0.036;(21.36±12.11)kU/L vs(11.28±10.78)kU/L, t=2.64,P=0.012].Patients experienced a decrease in NSE and CA19-9 concentrations following whole lung lavage[(39.20±10.24)μg/L vs(15.32±8.35)μg/L,t=8.02,P<0.05;(26.24±12.23)kU/L vs(18.84±5.64)kU/L,t=2.46,P<0.05].Immunohistochemical studies showed positive NSE and CA19-9 staining in lung biopsy specimen.Conclusion Elevated serum tumor markers including NSE, CA125 and CA19-9 would provide valuable clinical information to assess disease severity in silicosis.
6.Thrombocytopenia with absent radii syndrome: a case report.
Wen-ying ZHANG ; Chun-xiao FANG ; Tie-zhen YE
Chinese Journal of Pediatrics 2010;48(8):633-634
Humans
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Infant
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Male
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Radius
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abnormalities
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Syndrome
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Thrombocytopenia
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complications
7.Inhibitory effects of triptolide on cell proliferation and metastasis in Raji cells in vitro
Chun ZHANG ; Guohui CUI ; Fang LIU ; Qiuling WU ; Yan CHEN
Chinese Journal of Pathophysiology 1986;0(02):-
AIM:To investigate the inhibitory effects of triptolide on cell proliferation and metastasis in Burkitt's lymphoma cell line Raji cells.METHODS: The effects of triptolide on the growth of Raji cells were studied by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium(MTT) assay.The effects of triptolide on the cell apoptosis of Raji cells were detected by using Annexin Ⅴ/PI double-labled cytometry.The effects of triptolide on CXCR4 expression on Raji cells were studied by flow cytometric analysis.Chemotaxis assays were performed to observe the effects of triptolide on migration of Raji cells towards recombinant human SDF-1?(rhSDF-1?)in vitro.RESULTS: Triptolide inhibited the proliferation of Raji cells in a dose-and time-dependent way with a 24 h IC50 value of 43.06 nmol/L and a 36 h IC50 value of 25.08 nmol/L.Following the treatment of triptolide,the cell apoptosis rate was increased as the treatment concentration increased and the culture time extended.The effects were dose-and time-dependent.Triptolide could downregulate the expression of CXCR4 on Raji cells in a dose-dependent manner.Moreover,chemotaxis assay suggested that triptolide could block the migration of Raji cells to rhSDF-1? in vitro,and the inhibition was dose-dependent.CONCLUSION: Triptolide could inhibit the cell proliferation and induce the cell apoptosis of Raji cells.Furthermore,it could block the cell metastasis of Raji cells in vitro and the underlying mechanism might be related to the inhibition of the SDF-1/CXCR4 axis.
8.Aldosterone modulates the production of matrix metalloproteinase-2 and -9 via transforming growth factor β1 signaling pathway in podocytes
Zhan FANG ; Xifeng SUN ; Fangfang HE ; Zhonghua ZHU ; Chun ZHANG
Chinese Journal of Nephrology 2009;25(7):503-508
objective To assess the effect of aldosterone on the production of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9)and collagen Ⅳ in culture supematants of podocytes and the possible molecular mechanisms involved in the influence of aldosterone on the synthesis and degradation of extracellular matrix produced by podocytes. Methods Podecytes were treated with aldosterone at the concentration of 10-11, 10-9, 10-7 mol/L respectively. Cultured podocytes were examined at 24, 48 and 72 hours respectively. Spironolactone, a receptor antagonist of aldosterone, was added to observe the blocking effect on aldosterone. An inhibitor of TGF-β1 receptor was used to determine whether the effect of aldosterone on podocytes were mediated through TGF-β1 system. The enzymatic activities of MMP-2 and MMP-9 were assayed by gehtin zymography. Collagen Ⅳ 0.5 chain and TGF-β1 proteins released into culture supematants were assessed by Western blot and ELISA analysis. The adhesion rate of podocytes was monitored by flow cytometry. Results Aldosterone increased the activities of MMP-2 and MMP-9 in a dose- and time-dependent manner (P<0.05). Aldosterone decreased the level of collagen Ⅳ or5 chain protein in culture supernatants (P<0.05). Meanwhile, the expression of TGF-β1 was also increased (P<0.05). Spironolactone completely abolished the above-mentioned changes(P< 0.05). Blockage of TGF-β1 signaling with SB431542 prevented the aldosterone-induced upregulation of MMP-2 and MMP-9 as well as the downregulation of the collagen Ⅳ α5 chain protein and the adhesion rate of podocytes (P<0.05). Conclusions Aldosterone increases the activities of MMP-2 and MMP-9 but decreases the expression of collagen Ⅳ α5 chain and the adhension rate of podocytes possibly via TGF-β1 signaling pathway. Such alterations may contribute to glomerular podocyte injury associated with the GBM abnormality caused by the imbalance between matrix synthesis and degradation.
9.Interference of alkaline phosphatase on clinical cardiac troponin Ⅰ assay
Zejin LIU ; Fang ZHENG ; Zhenlu ZHANG ; Qingkun FAN ; Chun WANG
Chinese Journal of Laboratory Medicine 2010;33(9):831-833
Objective To evaluate the interference of ALP on cTnI assays. Methods One normal mixed plasma sample and 2 abnormal mixed plasma samples with different cTnI levels were prepared, and then divided them into 8 groups respectively. One group was randomly chosen as control while different amounts of ALP were added into the other seven groups. The concentrations of cTnI and ALP in each plasma portion were detected by ACCESS2 (Beckman-Coulter, Inc ) and AXSYM (Abbott Laboratories )separately. The results of the seven tested groups were then compared with those of the control, so as to evaluate whether ALP could interfere with the cTnI assay. Results When the chemiluminescent Access cTnI assay was carried out for detection of normal plasma, the concentration of ALP was up to 3 716 U/L and did not interfere with the test results of cTnI [(0. 04 ±0.01) μg/L] compared with those of the control portion [(0. 04 ± 0. 01 ) μg/L] (t = 0. 40, P > 0. 05 ). Once the concentration of ALP went beyond 917 U/L, the AXSYM cTnI assay results [( 0.08 ± 0. 01 ) μg/L] were higher than those of the normal control ( t =-4. 89, P<0. 01 ); When the concentration of ALP was up to 3 534 U/L, the test results of abnormal plasma cTnI detected by the Access assay [( 13.41 ±0. 17) μg/L] did not show significant differences from those of the control [(13.48±0.16) μg/L] (t=0. 52,P>0.05).Conclusions High concentration ofALP did not interfere with the Access cTnI assay or lead to false positive results. However, the high level of ALP( > 917 U/L) could interfere with the AXSYM cTnI assay and cause a false positive result.
10.Inter-arterial thrombolysis using rt-PA for acute cerebral ischemic stroke
Bing ZHANG ; Shaojun ZHENG ; Chun FANG ; Minghua LI
Journal of Interventional Radiology 2006;0(09):-
Objective To evaluate the safety and efficacy of intra-arterial thrombolysis in patients with acute cerebral ischemic stroke.Methods Twenty-one patients with acute internal carotid circulation infraction(internal carotid 3,MCA 12,ACA 5,lenticulostriate in 1) were treated with intra-arterial thrombolysis of recombinant tissue plasminogen activator(rt-PA) which was performed within 2-6 hours of symptom onset.Recanalization was observed during the operation.Intracerebral hemorrhage(ICH) was monitored immediately and 24 h after the treatment by CT or MRI scanning.Chinese stroke scale was used to evaluate the recovery of neurological functions pre-operatively and 30 d after the treatment.Results All the 21 patients were 100% success in receiving intra-arterial thrombolysis technique and revealed 16 having the degree of recanalization of 2 to 3 grade as regards to TMI,16 patients' degree of recanalization reached 2 to 3 grade according to TMI;5 patients showed 1 to 2 grade.Symptomatic ICH was observed in 3 patients,with two dead.Arterial dissection was found in one patient.Thirty days after the operation,17 patients' cerebral function reduced over 50 percent;2 less than 50 percent;and 2 died.The patients achieved 2 to 3 grade of recanalization were obviously getting better than those achieving 0 to 1 grade.Conclusions It is adapt to have intra-arterial thrombolysis with six hours from onset;but still have the danger of severe ICH.The treatment should be started as early as possible.