1.Effects of RNAi targeting CDX2 gene expression on biological characters of human leukemia cells and its relevant mechanisms
Weimei SUN ; Jianchang LI ; Xiuhong JIA ; Youjie LI ; Shenhua TANG
Tianjin Medical Journal 2016;44(6):679-682,683
Objective To detect the effects of siRNA targeting CDX2 gene expression on of BCR-ABL, caspase and Bax expressions, and the mechanisms thereof. Methods According to the earlier experiments, siRNA specifically targeting CDX2 gene (CDX2-siRNA) and the negative control sequence (CDX2-siRNA-NC) were selected, and then were transfected into K562 cells by Roche X-tremeGENE HP DNA Transfection Reagent. The flow cytometry analysis was used to detect the effects of siRNA on cell apoptosis. The expressions of BCR-ABL, caspase-9, Bax mRNA and protein were tested by RT-PCR and Western blot assay. Results MTT and flow cytometry analysis showed that after the silence of CDX2 gene expression, the proliferation of K562 cells was prohibited and the apoptotic rate of K562 cells was distinctly increased compared with that of normal cell group, but the negative control group had no significant change. According to the RT-PCR and Western blot assay, in comparison with the normal cell group and the negative control group, the expression levels of BCR-ABL mRNA and protein were obviously decreased, and the difference was statistic significance. On the other hand, the expressions of caspase-9 and Bax mRNA and protein were significantly higher than those of other two groups (P<0.05). Conclusion CDX2-siRNA can promote apoptosis of K562 cells obviously, and the mechanism is related with the down-regulation of BCR-ABL and the up-regulation of caspase-9 and Bax.
2.Proliferation and Immunophenotypic Analysis of CD3AK Cells Derived from Lymph Node
Shenglin MA ; Jianguo FENG ; Shenhua XU ; Lirong TANG ; Yutian LIN
Chinese Journal of Cancer Biotherapy 1995;0(02):-
Objective: To study the propagation and phenotypes changes of killer cell (CD3AK cell activated by CD3 mAb in vitro. Methods: Lymph nodes taken from lung cancer patient is dissociated into single cell suspension by mechanical method and cultured in culture medium added CD3 mAb and a little dose IL-2. We analyze cell immunophenotype by flow cytometry and proliferation by trypan blue exclusion test per 2 days. Results: Immunophenotypic analysis showed that CD3AK expressing CD3, CD8, CD56, CD25 increased, and reached a peak value which is 2.33 times than before culturing in the 8 th day. Conclusion: CD3 mAb added to the culture medium can obviously activate CD3AK cell and stimulate proliferation and keep its killer activity.
3.Diagnostic value of N-terminal pro-brain natriuretic peptide and MB isoenzyme of creatine kinase for heart failure in pneumonia children
Shenhua TANG ; Xiuhong JIA ; Jianchang LI ; Xiaomei LI ; Yanyan ZHANG
Journal of Clinical Pediatrics 2015;(8):694-697
ObjectiveTo investigate the diagnostic value of N-terminal pro-brain natriuretic peptide (NT-proBNP) and MB isoenzyme of creatine kinase (CK-MB) for heart failure (HF) in pneumonia children.MethodsThe NT-proBNP and CK-MB were assayed in 132 pneumonia children with HF, 138 pneumonia children without HF and 62 healthy children were recruited into this study. A receiver operating characteristics (ROC) curve and a logistic regression model were employed to assess the diagnostic accuracy of NT-proBNP and CK-MB for HF in pneumonia children.ResultsPneumonia children with HF had higher blood NT-proBNP and CK-MB than those in pneumonia children without HF and healthy controls (P<0.01 for both). Pneumonia children with HF had higher blood NT-proBNP and CK-MB than the pneumonia children without HF. The area under curves (AUCs) of NT-proBNP and CK-MB for HF were 0.85 and 0.72, respectively. The AUC for their combinational usage was 0.87.ConclusionBoth NT-proBNP and CK-MB are effective markers as diagnostic adjuncts for HF in pneumonia children. Combination of NT-proBNP and CK-MB can improve the diagnostic accuracy for HF in pneumonia children.
4.Influence on Migration of Vascular Endothelial Cells by Couplet Medicines of Reinforcing Qi and Activating Blood
Bingbing LI ; Shenhua YIN ; Decai TANG ; Wenhua ZANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1147-1152
This study was aimed to observe the influence on migration ability of human umbilical vein endothelial cell (HUVEC) in vitro by couplet medicines of reinforcing qi and activating blood, and initially screen medicines with relative obvious promoting or inhibiting effect on migration ability of HUVEC. The model of HUVEC cultured in vitro was established. The Paeonia, Ligusticum chuanxiong, Cortex moutan, Salvia, curcuma zedoaria, Sparganium, Astragalus, and ginseng were combined in pairs with the proportion of 1:2, 1:1, 2:1. There were 13 couplet medicines of reinforcing qi and activating blood. Serum pharmacological method was used to prepare medicated serum of the couplet medicines of reinforcing qi and activating blood. Scratch method was used to detect the effect of medicated serum of these couplet medicines of reinforcing qi and activating blood and activate blood herbs on the migration a-bility of HUVEC (with the density of 5í105/mL) after 24 hours. The results showed that compared with the blood serum of the blank group, the Cortex moutan group, Ligusticum chuanxiong group, Paeonia group, Salvia and Astra-galus (1:2) group, Salvia and Astragalus (1:1) group, Ligusticum chuanxiong and Astragalus (1:2) group had obvious promoting effect on the migration ability of VEC (P < 0.05 or P < 0.01). The Sparganium group, curcuma zedoaria group, Cortex moutan group, curcuma zedoaria and Astragalus (2:1) group, Sparganium and ginseng (2:1) group, Spar-ganium and Astragalus (1:1) group had obvious inhibiting effect on the migration ability of VEC (P< 0.05 or P<0.01). It was concluded that different compatibility of medicines of reinforcing qi and activating blood and activate blood herbs had different promoting or inhibiting effect on migration ability of HUVEC. It may be related to the mechanism of action of these drugs on promoting or inhibiting angiogenesis at the cellular level.
5.Influence of Qi-reinforcing and Blood-activating Herbs on Protein Expression and mRNA Expression of SDF-1 and CXCR4 in Infarcted Myocardium Edge Area of Acute Myocardial Infarction Rat Model
Wenhua ZANG ; Bingbing LI ; Decai TANG ; Shenhua YIN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(6):1377-1383
This study was aimed to observe the influence of qi-reinforcing and blood-activating(QRBA) herbs onan-giogenesis of the myocardial microvascular, SDF-1 and CXCR4 protein expression as well as mRNA expression in the infarcted myocardium edge area of acute myocardial infarction (AMI) ratmodel. The AMI rat model was estab-lished. The immunohistochemical staining method was used in the detection of vWF protein expression in the myocar-dial tissues. The MVC account was recorded. The SDF-1 factor and its specific receptor factor CXCR4 were detected by the western blot and realtime-PCR technique in the infarcted myocardium edge area of rats from each group. The results showed that in the new generated microvessels which were staining marked by the vWF factor can be seen in infarcted myocardium edge area of rats from the sham-operated group, model group, each medication group. The new generated microvessels in the myocardium of rats in the sham-operated group were not obvious. Small amount of new generated microvessels can be seen in rats from the model group. More new generated microvessels can be seen in rats from each medication group. The comparison between the model group and the sham-operated group showed sta-tistical difference (P<0.05). The comparison between each medication group and the model group showed statistical difference(P<0.05 or P<0.01). Compared with the sham-operated group, SDF-1, CXCR4 and mRNA expression were obviously increased in the myocardium of rats in the model group (P<0.05 or P<0.01). Compared with the model group, SDF-1, CXCR4 protein and mRNA expression were obviously increased in the myocardium of rats from each medication group (P<0.05 or P<0.01). It was concluded that herbs such as Salvia, couplet herbs of Salvia and Astra-galushad stimulation effectonangiogenesis. Mechanism of these drugs in angiogenesismay be through the promotion of SDF-1 and CXCR4 protein as well as mRNA express.