ABSTRACT
Objective To investigated the effect of procyanidin (PC) on the expression of cysteine proteinase -3 (Caspase -3) in type 2 diabetes mellitus SD rats with focal cerebral ischemia. Methods Following the random principle, 40 healthy Sprague - Dawley (SD) rats were numbered sequentially and randomly divided to normal rats with focal cerebral ischemia group,type 2 diabetes mellitus SD rats with focal cerebral ischemia group,PC low/ middle/ high -dose groups,with 8 rats in each group. The type 2 diabetes mellitus - MCAO model was set up. The doses of PC for low,middle and high - dose groups were 50 mg/ kg,100 mg/ kg,200 mg/ kg. Immunohistochemistry method was used to measure the activity of Caspase - 3. Results Compared with that in the normal rats with focal cerebral ischemia group[(11. 42 ±2. 52)],the expression of Caspase -3 increased in the type 2 diabetes with ischemia group[(15. 00 ± 2. 38)](t = 2. 17,P < 0. 01). Compared with that in the type 2 diabetes with ischemia group,the expression of Caspase - 3 decreased in the PC groups[(9. 38 ± 2. 00),(7. 71 ± 1. 55),(6. 96 ± 1. 57)](t = 2. 86,3. 13,3. 36,all P < 0. 01),whereby the middle and high - dose groups showed more significant decrease (t = 1. 92,2. 03,all P <0. 01) and with no statistically significant difference between the two groups(t = 1. 13,P > 0. 05). Conclusion PC can decrease the expression of Caspase - 3 protein in type 2 diabetes mellitus SD rats with focal cerebral ischemia, finally may inhibit the apoptosis.
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Objective To examine expression levels of autophagy gene pULK and PI3KC3 and to explore their correla?tion with non-small cell lung cancer (NSCLC). Methods A total of 77 samples of surgical resection from NSCLC speci? mens (including 31 cases of squamous cell carcinoma, 31 cases of adenocarcinoma and 15 cases of large cell undifferentiated carcinoma) and 21 samples of same normal lung tissue were randomly selected. Expressions of pULK and PI3KC3 in lung tissues were assessed by immunohistochemistry and Western blot. All dates were analyzed using SPSS13.0 statistical pack?age. Results Immunohistochemistry indicated that pULK and PI3KC3 localized into the cytoplasm. The expression levels of pULK and PI3KC3 are significantly lower in patient with NSCLC than those in peri-tumor tissue (35.1%vs 81.0%, 40.3%vs 76.2%respectively, P<0.01) . Immunohistochemistry and Western bolt analysis confirmed that pULK and PI3KC3 ex?pressions were significantly down-regulated (P<0.01) in patients with low grade cellular differentiation, metastasis of lymph node, or stageⅢandⅣ. And expression levels of pULK and PI3KC3 in NSCLC did not differ significantly with ages, gen?der, tumor size and pathological type. Correlation analysis showed that the expression of PI3KC3 was positively correlated with pULK. Conclusion pULK and PI3KC3 expressions were lower in NSCLC than those in normal lung tissue group. The expression levels of pULK and PI3KC3 in NSCLC were correlated with patient's clinical stage, differentiation grade, lymph node metastasis but were unrelated with age, gender, histological type and size.
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Objective To compare different ways to trace bone marrow mesenchymal stem cells (BMSCs) after being transplanted in cerebral ischemia-reperfusion injury. Methods Male SD rats of SPF grade were randomly divided into sham group, model group (ischemia-reperfusion,IR), BrdU tracing group, PKH26 tracing group and GFP tracing group. Fo?cal cerebral ischemia-reperfusion model was established by blocking middle cerebral artery. 24 hours after cerebral isch?emia-reperfusion injury, 10μL BMSCs that were labeled respectively by BrdU, PKH26, GFP were added respectively into BrdU, PKH26 and GFP tracing group while equal volum of normal saline was added into sham group and model group. Mod?el and transplanting cells efficacy was determined by neural behavioral score, TTC staining and brain water content;Neurons were counted using tar violet staining;The number of transplant cells in the transplanting site was assessed by fluorescence microscopy. Results Before transplanting, there was no significant difference among BrdU, PKH26 and GFP group in cell labeled efficacy. By contrast, neural behavioral score, brain infarct volume and brain tissue water content were significantly lower in all three tracing groups than that in model group 4 weeks after transplantation while neuron counts were markedly higher. There was no significant difference of above parameters among the three tracing groups. However, the number of traced transplanting cells in damaging area in GFP group is significantly higher than that in BrdU group and PKH26 group. Conclusion In cerebral ischemia-reperfusion injury, the tracing effect of GFP last longer, therefore it is significantly more effective than BrdU and PKH26.
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Objective To investigate the effects of zinc fingers and homeoboxes 3 (ZHX3) silence on expressions of smad3, smad4 and RUNX2 in bone marrow mesenchymal stem cells (BMSCs). Methods ZHX3 low expression vector (ZHX3 silent group) was constructed and was transfected to rat BMSCs. Empty vector was transfected into BMSCs and was used as vehicle control group, and wild type BMSCs was used as the control group. The cell transfection rate was measured under a fluorescence microscope, and then the successful transfection was identified. The immunocytochemistry and immu?noblotting methods were used to detect the expression levels of smad3, smad4 and RUNX2. Results (1) Cells with BMSCs phenotype can be obtained by recovery culturing. (2) After transfection, the green fluorescent protein was found in ZHX3 si?lence group and vehicle control group. Blank control group showed no significant fluorescence. The expression level of ZHX 3 was significantly lower in ZHX3 silence group than that of vehicle control group. (3) Results of immunofluorescence asssay showed that the positive expressions of smad3 and smad4 were located in nucleus and cytoplasm, the positive expression of RUNX2 was mainly located in nucleus. Positive cells were observed in three groups. There was no significant difference in fluorescence intensity between the control group and the vehicle control group, but the fluorescence intensity was significant?ly lower in ZHX3 gene silence group than that of two control groups. (4) There were no significant differences in expressions of smad3, smad4 and RUNX2 betweem control group and the vehicle control group, but they were significantly higher than those of ZHX3 silence group(P < 0.05). Conclusion ZHX3 gene silence can delay vitro osteogenesis of BMSCs, which may play a role by the down-regulated expression levels of smad3, smad4 and RUNX2.