1.Analysis on value of combined detection of Hcy,hs-CRP and Lp-PLA2 in diagnosis of ischemic cerebral stroke
Meng DING ; Yingwei WU ; Shoujun XIE ; Xueli GENG ; Yaju WANG
International Journal of Laboratory Medicine 2017;38(14):1898-1900
Objective To analyze the significance of levels and combined detection of plasma homocysteine(Hcy),high-sensitivity C reactive protein(hs-CRP) and lipoprotein-associated phospholipase A2(Lp-PLA2) in patients with ischemic cerebral stroke(ICS).Methods A total of 140 patients with ICS were selected into ICS group,and 100 healthy individuals were recruited in the control group.The plasma levels of Hcy,hs-CRP,Lp-PLA2 were examined,then the positive rates of them were statistically analyzed.Analyzed the value of the levels and joint detection of Hcy,hs-CRP and Lp-PLA2.Results The plasma levels and the positive rates of Hcy,hs-CRP and Lp-PLA2 in ICS group were statistically higher than those in the control group(P<0.05).The area under the receiver-operating characteristic curve of combined detection of Hcy,hs-CRP and Lp-PLA2 in diagnosis of ICS was 0.949,the sensitivity was 0.850,and the specificity was 0.807,which were higher than Hcy,hs-CRP and Lp-PLA2 alone.Conclusion The plasma levels of Hcy,hs-CRP and Lp-PLA2 increase in ICS patients,and the combined detection of the three indicators might improve the early diagnostic value of ICS.
2.RNA interference affects the feak-headbox 3a gene expression in myoblast cell line L6
Jie DING ; Bingsheng LIANG ; Zhifeng DA ; Zhixiang ZHU ; Jian WEI ; Yingwei JIA ; Yong FENG
Chinese Journal of Tissue Engineering Research 2013;(33):5974-5980
BACKGROUND:Recent studies found that some factors play important role in the process of denervated muscle atrophy, especial y the feak-headbox transcription factor, is the key element to regulate the denervated muscle atrophy.
OBJECTIVE:To investigate the effect of RNA interference on inhibiting feak-headbox 3a gene expression in vitro.
METHODS:The myoblast cel line L6 were cultured in the 6-wel cel culture plates, then pEGFP-N1 and smal
interfering RNA recombinant plasmid with the same ratio was transfected under the Lipofectamine2000 mediation to optimize the transfection efficiency of the detection system;2μg smal interfering RNA recombinant plasmid of feak-headbox 3a gene were transfected with myoblast cel line L6 for 48 and 72 hours.
RESULTS AND CONCLUSION:At 48 hours after pEGFP-N1 and siRNA recombinant plasmid transfection, a large number of bright green fluorescent displayed under fluorescence microscope with higher transfection efficiency. Real-time quantitative PCR analysis showed that there were significant differences in the sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (P<0.05), and the inhibition effect was more significant at 72
hours after transfection when compared with that at 48 hours after transfection. Western Blot gray analysis showed that there were significant differences in sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ,
feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after
trasfection (P<0.05), and the inhibition effect was more significant at 72 hours after transfection when compared with that at 48 hours after transfection, which was same with the effect on mRNA level. RNA interference in vitro can
significantly inhibit the fork-head transcription factor feak-headbox 3a gene expression, and the inhibition effect of feak-headbox 3a gene smal interfering RNA recombinant plasmid transfected with the sequence on the mRNA and protein level of feak-headbox 3a is not clear, which can provide new idea for the gene therapy of RNA mediated denervated skeletal muscle atrophy.
3.Tim-3 promoted renal ischemia/reperfusion injury through regulating mononuclear phagocyte system function
Yunshan GUO ; Yaohai DING ; Yingwei ZHANG ; Xiaoyu JIANG ; Hongdong LI ; Zhen LI ; Moyan LIU ; Yi LIU
Chinese Journal of Organ Transplantation 2018;39(2):109-115
Objective To investigate the effects of Tim-3 on the renal ischemia-reperfusion injury (IRI),and explore the role of monocyte-macrophage cell system.Methods Totally 72 C57BL/ 6 mice were randomly divided into four groups (n =18 each).(1) IR + Tim-3 rnAb group (experimental group):Each mouse was intraperitoneally injected with 200μg of anti-Tim-3 mAb and the IR model of mouse kidney was established after 1 day;(2) IR + IgG monoclonal antibody group (negative control group):each mouse was intraperitoneally injected with anti-IgG mAb (200 μg) and the IR model of mouse kidney was established after 1 day;(3) IR group:mouse kidney IR model was established only;(4) Control group:mouse kidney IR model was not established.At 6,24 and 48 h after IR respectively,venous blood of 6 mice in each group was taken from the infrarenal vein.Scr and CystinC were detected and PAS staining was used to observe the pathological change of renal tissues.Cell apoptosis was detected by TUNEL staining.Pax,bcl-2 and caspase-3 expression in renal tissue was detected by Western blotting.Immunohistochemistry was used to detect the distribution of Tim-3 and activated macrophage cells.Flow cytometry and ELISA were used to evaluate the level of Tim-3 and inflammatory cytokines secretion respectively.Results Compared with control group,the Tim-3 expression was dramatically increased in IR group and I/R + Tim-3 mAb group.The serum Scr and CystinC levels were increased in IR group,and Tim-3 blocking decreased the levels of serum Scr and CystinC (P<0.05).PAS and TUNEL staining showed that renal injury score and apoptotic index were higher in IR group than those in control group.Tim-3mAb significantly decreased those markers,and ameliorated the renal tubulointerstitial injury induced by IRk The expression levels of Caspase-3 and Bax/bcl-2 was increased in IR group,but deceased by Tim-3mAb.IR induced F4/80 + distribution and inflammatory cytokines secretion in renal tubular interstitial tissues,while Tim-3mAb down-regulated F4/80 + activation and the levels of inflammatory cytokines.Conclusion The findings demonstrated Tim-3 may promoted renal IRI through regulating mononuclear phagocyte system function.
4.The protective effect of bone marrow mesenchymal stem cells carrying antioxidant gene superoxide dismutase on paraquat lung injury in mice.
Hong LIU ; Yingwei DING ; Yuehui HOU ; Guangju ZHAO ; Yang LU ; Xiao CHEN ; Qiqi CAI ; Guangliang HONG ; Qiaomeng QIU ; Zhongqiu LU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2016;34(1):1-7
OBJECTIVETo explore the possible mechanism and protective effect of BMSCs (bone mesenchymal stem cells) carrying superoxide dismutase (SOD) gene on mice with paraquat-induced acute lung injury.
METHODSTo establish the cell line of BMSCs bringing SOD gene, lentiviral vector bringing SOD gene was built and co-cultured with BMSCs. A total of 100 BALB/c mice were randomly divided into five groups, namely Control group, poisoning group (PQ group) , BMSCs therapy group (BMSC group) , BMSCs-Cherry therapy group (BMSC-Cherry group) , BMSCs-SOD therapy group (BMSC-SOD group) . PQ poisoning model was produced by stomach lavaged once with 1 ml of 25 mg/kg PQ solution, and the equal volume of normal saline (NS) was given to Control group mice instead of PQ. The corresponding BMSCs therapy cell lines were delivered to mice through the tail vein of mice 4h after PQ treatment.Five mice of each group were sacrificed 3 d, 7 d, 14 d and 21 days after corresponding BMSCs therapy cell lines administration, and lung tissues of mice were taken to make sections for histological analysis. The serum levels of glutathione (GSH) , malondialdehyde (MDA) , SOD, and the levels of transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α) in lung tissue were determined. The level of SOD was assayed by Westen-blot.
RESULTSCompared with Control group, the early (3 days) levels of SOD protein in lung tissue of PQ group obviously decreased, and the late (21 days) levels of SOD obviously increased, while in therapy groups, that was higher than that in PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) . Compared with Control group, the levels of plasma GSH and SOD of PQ group and each therapy group wae significantly lower than those in Control group, while in therapy groups, those were higher than those of PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) .Compared with Control group, the level of plasma MDA, TNF-α and TGF-β in PQ group and therapy groups were significantly higher, while in therapy groups, that was lower than that in PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) . Lung biopsy showed that, the degree of lung tissue damage in each therapy group obviously reduced.
CONCLUSIONSOD is the key factor of the removal of reactive oxygen species (ROS) in cells, that can obviously inhibit the oxidative stress damage and the apoptosis induced by PQ, thus significantly increasing alveolar epithelial cell ability to fight outside harmful environment.
Acute Lung Injury ; chemically induced ; therapy ; Animals ; Antioxidants ; metabolism ; Cell Line ; Glutathione ; blood ; Lung ; pathology ; Malondialdehyde ; blood ; Mesenchymal Stem Cell Transplantation ; Mice ; Mice, Inbred BALB C ; Oxidative Stress ; Paraquat ; poisoning ; Superoxide Dismutase ; blood ; genetics ; Transforming Growth Factor beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism