Insulin induced LPL mRNA expression in THP-1 macrophage and acceleration of transferring the macrophage to foam cell

AIM:To investigate the effect of insulin on ox-LDL transferring the THP-1 cells to foam cells and influencing the LPL mRNA expression in THP-1 cells.METHODS:THP-1 cells were incubated with 50 mg/Lox-LDL and insulin at concentrations of 10 mU/L, 100 mU/L, 1 000 mU/L and 10 000 mU/L, respectively. The expression of LPL mRNA in cells was detected by RT-PCR. Lipoprotein lipase of THP-1 cells was presented by no-specific lipase staining. THP-1 cells were stained with oil red O. Accumulation of total cholesterol (TC) in THP-1 cells was determined with oxidase assay.RESULTS:In 100 mU/L、1 000 mU/L、10 000 mU/L insulin groups, LPL mRNA expression increased 2 times, the average cell perilength was longer, the percentage of positive oil red O staining cells was significant higher, the content of cholesterol in THP-1 cells was higher than in ox-LDL control (P＜0.05).CONCLUSION:Insulin accelerates transferring of THP-1 cells to foam cell with exposed to ox-LDL because LPL mRNA expression increased in the cells.     

Bcl-2 antisense oligodeoxynucleotide increases the sensitivity of HL60 and K562 cells to daunorubicin

AIM:To investigate whether the bcl-2 antisense oligonucleotide increases the sensitivity of HL60 and K562 cell lines to daunorubicin.METHODS:IC50 for HL60 and K562 was determined with MTT method, the expression levels of Bcl-2 protein were assayed by immunofluorescence using fluoresce isothiocyanate labeling. In addition, apoptosis was detected by morphological observation and flow cytometric analysis of DNA fragmentation.RESULTS:It was found that the two oligonucleotides directed against the coding region and the translation initiation of bcl-2 mRNA, combined respectively with daunorubicin, inhibited expression of bcl-2 protein, increased apoptosis in HL60 and K562 cells, and decreased IC50 of daunorubicin significantly (P＜0.05). Compared to the antisense oligonucleotide directed against the translation initiation of bcl-2 mRNA, the antisense oligonucleotide directed against the coding region showed stronger effects in the aspects of increasing the sensitivity of HL60 cells to daunorubicin (P＜0.05).CONCLUSIONS:These two antisense sequences in the translation initiation and the coding region of bcl-2 mRNA increased the sensitivity of HL60 and K562 cell lines to daunorubicin in a sequence-specific manner.     

Biological characteristics of long passaging rat mesenchymal stem cells

AIM:To investigate multi-potential of rat bone marrow mesenchymal stem cells (rBMMSC) and mutation inclination, the rBMMSC were long passaged in vitro. METHODS:Cellular cycles of different passages were assayed by FACSan flow cytometry and karyotypes of passage 6, passage 25 and passage 45 were compared by G-binding analysis. RESULTS:The early passages and long-term passages all showed strong proliferation; passage 6, passage 25 and passage 45 all showed normal karyotype. CONCLUSION:Long-term culture and passage of rBMMSC still remains strong proliferation. With this capability, the mutation inclination is not enhanced.     

Effect of tea-polyphenols on the activation of NF-κB and the expression of TGF-β1 mRNA in THP-1 cells incubated with VLDL,LDL or ox-LDL

AIM:To investigate the effect of tea-polyphenols (TP) on the activation of NF-κB and the expression of TGF-β₁ mRNA in THP-1 cells (a human acute monocytic leukemia cell line). METHODS:THP-1 cells were incubated with the different concentrations of TP, VLDL, LDL or ox-LDL. In the THP-1 cellls, the nuclear malposition rate of NF-κB was detected with immunohistochemistry technique, the positive index of the TGF-β₁ mRNA expression was detected by hybridization in situ, and accumulation of total cholesterol (TC) in cells incubated with 0.4-40 μg/L TP was determined with oxidase assay. RESULTS:The nuclear malposition rate of NF-κB, the positive index of the TGF-β₁ mRNA expression and TC in THP-1 cells incubated with 0.4-40 μg/L of TP were lower than those with 0 μg/L of TP in TP-V group, TP-L group and TP-O (P＜0.05). The differences of these markers in THP-1 cells incubated with more than 40 μg/L TP in TP-V group, TP-L group and TP-O were not statistically significant, compared with TP-C group (P＞0.05). CONCLUSION:TP inhibited the activation of NF-κB, the expression of TGF-β₁ mRNA and the foam cell formation in the mono-macrophage.     

姬松茸与双孢蘑菇不同菌株的RAPD扩增研究

以4个姬松茸菌株和2个双孢蘑菇菌株为材料，用20个随机引物对它们进行RAPD扩增，通过对供试菌株遗传相似系数的估算和系统聚类分析，构建姬松茸和双孢蘑菇遗传相关树状聚类图谱，结果表明，进行RAPD扩增的20个随机引物中，有12个能产生三种带型，当遗传相似系数升至0.8713时，这6个菌株被分为3类，第一类为菌株18和26，第二类为菌株13和12，第三类为菌株33和45。     

A reactive pattern of ischemia-induced nestin protein in the rat brain

AIM:To explore the expressive profile of nestin protein in the focal ischemic brain and to study the recovery mechanism of brain focal infarct.METHODS:Cellular morphology,time-course and distribution pattern of nestin positive response were immunohistochemically examined in different brain regions of 36 adult male SD rats. RESULTS:Nestin positive response of different brain regions in sham operated rats was present in small- and micro-vasculartures and the third ventricle bottom and ependyma. A large number of nestin positive cells were detected in ischemic brain, and were more remarkable in the cortical areas of parietal lobe and preoptic area as well as ischemic caudoputamen. Stellate nestin positive cells were located in the deep layer of ischemic cortex, but fibrillary cells were located in the shallow layer. Nestin positive cells in the ischemic caudoputamen showed the same changes of morphology as those cells in the deep layer of ischemic cortex. Morphological and number alterations of nestin positive cells were the most remarkable at 1 weeks post-ischemia, which showed more hypertrophy and proliferation in morphology, and a marked increase in number was present in the ischemic cerebral cortex and the ischemic caudoputamen. These alterations of nestin positive cells persisted up to 6 weeks post-ischemia, and then, the nestin positive response in the ischemic brain decreased gradually.CONCLUSION:Focal cerebral ischemia induces nestin re-expression on reactive astrocytes, which may be very important to the self-recovery of cerebral infarct.     

Effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts

AIM: To investigate the effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts. METHODS: In the present experiment, the human vascular fibroblasts were cultured and effects of platelet-derived growth factor-BB on DNA and collagen protein synthesis in human vascular fibroblasts were observed by using [³H]-TdR incorporation and [³H]-proline incorporation in vitro. RESULTS: Platelet-derived growth factor-BB significantly promoted NDA synthesis and collagen protein synthesis of quiescent human vascular fibroblasts, with a maximal response at a concentration of 30μg·L^-1at 24 h and 36 h, respectively. CONCLUSION: Platelet-derived growth factor-BB promotes DNA and collagen protein synthesis in cultured human vascular fibroblasts.     

Effect of 4-chloro-2-methylphenoxyacetic acid and 2,4-dimethylphenol on human erythrocytes

The effects of exposure of human erythrocytes to different concentrations of 4-chloro-2-methylphenoxyacetic acid (MCPA) and its metabolite—2,4-dimethylphenol (2,4-DMP) were studied. The investigations concerned mainly the content of glutathione (GSH and GSSG), glutathione peroxidase (GSH-Px), glutathione transferase (GST), and the level of adenine energy charge (AEC). Reactive oxygen species (ROS) such as hydroxyl radical, superoxide anion, hydrogen peroxide, and nitric oxide are produced during normal processes in the cell. Under normal conditions, antioxidant systems of the cell minimize damage caused by ROS. When ROS generation increases to an extent that it overcomes the cellular antioxidant systems, the result is oxidative stress. We observed that MCPA and 2,4-DMP decreased the level of GSH in erythrocytes in comparison with control. MCPA did not affect glutathione peroxidase and glutathione transferase activity, while 2,4-DMP increased their activity. 2,4-DMP decreased the level of ATP and increased the content of ADP and AMP, leading to the fall of the level of AEC. MCPA and 2,4-DMP transform hemoglobin into methemoglobin, thus preventing oxygen transport. Comparison of the toxicity of MCPA and 2,4-DMP revealed that the most prominent changes occurred in human erythrocytes incubated with 2,4-DMP.     

黄花蒿粗提物对几种害虫拒食性的初步研究

以黄花镐为原料进行浸提，经生物活性测定，结果表明黄花蒿Artenisia annua L.粗提物对供试的6种害虫均具有拒食性。其中对黑翅土白蚁Odontotermes formosanus Shiraki、赤拟谷盗Thibolium castaneum Herbst、谷蠹Rhizopertha dominica Fabricius拒食性极强，对棉蚜Aphis gossypii Glover、棉红蜘蛛Tetranychus urticae Koch及豇豆荚螟Etiella zinckenella Treitschke 也具有较强的巨食性。使用黄花蒿粗提物时，以稀释500倍和800倍效果最好，处理与对照之间有极显著差异。     

南方灭鼠应用稻谷与大米毒饵的比较

生物测定法测得谷壳对复方88-1灭鼠剂的阻滞率为54．3％，光谱分析法测得其对敌鼠钠盐的阻滞率在47．43％—57．00％之间。相同条件下，褐家鼠Rattus norvegicus取食敌鼠钠盐稻谷毒饵量为大米毒饵的2倍多。在南方潮湿的环境下，大米毒饵更易松散、霉变，使其适口性更低。综合考虑认为南方灭鼠使用稻谷毒饵更为合适。