Effects of oleanolic acid on apoptosis and cell cycle of HL-60 cells in vitro

AIM: To study the mechanism of oleanolic acid induced apoptosis and its influence on cell cycle in HL-60 cells in vitro. METHODS: The HL-60 cells were treated with different concentrations of oleanolic acid and then cultured for 12 h, 24 h, 48 h and 72 h, respectively. MTT assay was used to evaluate the inhibitory effect of oleanolic acid on HL-60 cells in vitro. The argarose gel electrophoresis was used to detect the chromatin DNA fragmentation. FACS was used to analyze the cell cycle of HL-60 cells. Western blotting was used to detect the activation of caspase-3 which has been confirmed the last execution of apoptosis pathway. RESULTS: MTT assay showed that oleanolic acid dramatically inhibited the growth of HL-60 cells in vitro, more than 50% HL-60 cells were inhibited when the cells were treated with 80 μmol/L oleanolic acid for 48 h; the apparent DNA ladder was detected after exposure of HL-60 cells to oleanolic acid for 48 h. FACS analysis showed that cell cycle of HL-60 cells was arrested in G₁ phase, the inhibition ratio of HL-60 cells achieved 63.24% and 67.90% after treated with oleanolic acid for 48 h and 72 h correspondingly. Western blotting detected the activation of caspase-3 after exposure of HL-60 cells to 80 μmol/L oleanolic acid for 48 h. CONCLUSION: These results suggest that oleanolic acid induces apoptosis and the cell cycle of HL-60 cells is arrested in G₁ phase.     

栽培条件对半夏叶片POD、SOD、CAT活性的影响

为了筛选能提高半夏叶片POD、SOD、CAT活性的栽培条件,用愈创木酚法、NBT光化还原抑制法和紫外分光光度法分别测定了不同基质不同施肥条件下半夏叶片中过氧化物酶（POD）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性。结果表明：半夏叶片POD酶活性受基质影响不明显,但不同施肥处理对其活性影响极显著,其中施用1/2MS+K+P（KH2PO4 100 mg/L）的处理半夏叶片POD酶的活性最强;半夏叶片中SOD和CAT酶的活性受不同基质的影响比较明显,其中以珍珠岩为基质栽培的半夏叶片中两种酶的活性均显著高于以土壤为基质栽培的半夏;不同施肥处理对这两种酶的活性也有影响,其中施用1/2MS+K+P（KH2PO4 100 mg/L）的处理半夏叶片SOD酶的活性最强。栽培半夏以珍珠岩为基质、施用1/2MS+K+P（KH2PO4 100 mg/L）的处理对于提高半夏叶片中POD、SOD、CAT 3种同工酶的活性为佳。     

铜绿假单胞菌生物膜肺部感染大鼠血清IgG抗体及肺部IFN-γ水平的变化

目的 了解人工铜绿假单胞菌(PA)生物膜肺部感染对机体免疫功能的影响.方法 由支气管内直接注入PA(菌种为PAO579)藻酸盐微粒1×109CFU/mL),建立慢性PA生物膜肺部感染大鼠模型,以支气管内注入生理盐水作为对照,2周后评估血清PA特异性抗体IgG水平、肺部IFN-γ反应的变化及其与肺组织病理学的相关性.结果...     

Response of common carp (Cyprinus carpio) larvae to different dietary levels and forms of supply of medium-chain fatty acids

The effects of dietary medium-chain fatty acids (MCFA) on survival, growth and fatty acid composition of first-feeding carp larvae were investigated in two 21-day feeding trials. In trial 1, one diet CO was supplemented with 10% coconut oil, two diets, M1 and M2 with 1.7 or 4.2% interesterified medium-chain triacylglycerols (MCT), and two other diets T1 and T2 with 1% tricaprylin and 0.7% tricaprin, or 2.5% tricaprylin and 1.7% tricaprin. The five diets were made isolipidic (18% of dry diet) with triolein and compared to a control diet without MCFA. With a low dietary level (1% of dry diet) of caprylic acid (C8:0), final survival and mean weight of larvae fed CO, M1 and T1 were not significantly different from those of control (68 ± 6% and 62 ± 8 mg, respectively). In contrast, with a high dietary C8:0 level (2.5% of dry diet), final survival and mean weight of larvae fed M2 and T2 were markedly decreased (23 ± 2% and 40 ± 8 mg, respectively), without effect of the form of supply. In trial 2, larvae were fed six diets with graded tricaprylin contents (from 0 to 10% in diet). Survival was not significantly affected but growth was decreased by C8:0 levels higher than 2% of dry diet. Fatty acid composition of larval total lipids revealed high levels of lauric and myristic acids in larvae fed coconut oil. Deposition of C8:0 and capric acid (C10:0) was low after MCT feeding, but depended of the form of supply, being higher with pure than with interesterified MCT. Relatively high amounts of C8:0 and C10:0 were recorded in total lipids of larvae fed 10% tricaprylin. It is concluded that high levels of caprylic acid decrease growth but that low levels are well utilised by carp larvae, irrespective of the form of supply.     

FTIR and Solid State13C NMR Spectroscopy of Proteins of Wet Cooked and Popped Sorghum and Maize

Fourier transform infrared (FTIR) and solid state¹³C NMR spectroscopic methods were used to investigate changes in maize and sorghum proteins on wet cooking and popping. FTIR spectra indicated that wet cooking led to proteins in two normal sorghums, namely NK 283 (a red hybrid) and KAT 369 (a white variety), two sorghum mutants (P850029 and P851171) and a maize hybrid (PAN 6043) assuming more antiparallel intermolecular β-sheet character, possibly at the expense of some α-helical conformation. Solid state¹³C NMR, using the technique of Cross Polarisation Magic Angle Spinning showed shifts of the protein carbonyl carbon and α-carbon resonances upfield on wet cooking in all samples, also indicating a change in protein secondary structure from α-helical to β-sheet conformation. The extent of secondary structural change on wet cooking seemed to be greater in sorghum than in maize and may have a bearing on the inferior protein digestibility of wet cooked sorghum compared to maize. Popping produced the same secondary structural change as observed for wet cooking in both sorghum and maize. However, the extent of change on popping was less than on wet cooking in sorghum and maize.     

高校图书馆服务工作中的知识产权问题

就高校图书馆服务工作中存在的知识产权问题进行了探讨, 分析问题产生的原因, 并就如何解决及避免高校图书馆服务中的知识产权问题提出了相应的对策。     

硬铝合金半连续铸造的裂纹缺陷分析

对凝固过程中流场、应力场、温度场及微观组织形态进行数值模拟,能帮助工艺设计人员分析不同时刻凝固过程的温度分布、金属流态、结晶晶粒大小、应力分布等重要物理参数,从而预测疏松、偏析、夹杂及热裂纹等缺陷。     

小鼠B7-H4原核表达载体的构建、表达及多克隆抗体的制备

目的探讨小鼠B7-H4原核表达载体的构建、表达及多克隆抗体的制备。方法采用特异性引物扩增小鼠B7-H4（mB7-H4）胞外功能区DNA,经酶切、拼接构建原核表达载体pET28a-mB7-H4。将构建的重组表达质粒转化入E.coliBL21（DE3）菌株,采用IPTG诱导表达、Ni-NTA柱亲和层析纯化目的蛋白、SDS-PAGE分析蛋白纯度。将纯化的目的蛋白免疫家兔制备多克隆抗体,并对其进行纯化及鉴定。结果序列测定证实了构建的pET28a-mB7-H4重组表达载体含有mB7-H4编码序列,其序列分析与GenBank中公布序列对比一致,质粒在E.coli中诱导表达相对分子质量（Mr）为26.5 KD的目的蛋白,SDS-PAGE分析表明纯化后的目的蛋白达到电泳纯。双向琼脂扩散法检测抗体效价为1∶16,ELISA法检测抗体效价为1∶12 800,Western blot分析显示抗体能特异性结合mB7-H4。结论成功制备了高表达重组mB7-H4蛋白的原核表达载体及高效价抗mB7-H4多克隆抗体,为进一步研究B7-H4的功能奠定了基础。     

Schisandra total lignin attenuates apoptosis of endoplasmic reticulum pathway to delay mouse brain aging

AIM:To investigate the role of schisandra total lignin (SCL) in anti-aging of the mouse brain. METHODS:A D-galactose induced mouse aging model was established. The following groups in this study were set up: control group, model group, SCL low dose group ［SCL (L)］, SCL moderate dose group ［SCL (M)］ and SCL high dose group ［SCL (H)］. Learning and memory abilities were measured by mouse jumping experiments. The expression of ubiquitin (Ub), glucose-regulated protein 78 （GRP78）, protein disulfide isomerase (PDI), CCAAT /enhancer-binding protein homologous protein (CHOP), Bcl-2 and Bax proteins was detected by Western blotting. In addition, the protein expression of Bcl-2 and Bax in the aging cerebral cortex was also observed by immunohistochemistry. RESULTS:In learning test, compared with control group, the number of errors within 5 min increased in model group (P<0.05). Compared with model group, the number of errors within 5 min decreased in SCL (L) group, SCL(M) group and SCL(H) group (P<0.05). In memory test, compared with control group, incubation period of the first jumping off the platform was shorter and the number of errors within 5 min increased in model grou(P<0.05). Compared with model group, the incubation period of the first jumping off the platform was longer and the number of errors within 5 min decreased in SCL (L) group , SCL(M) group and SCL(H) group (P<0.05). Compared with control group, the protein expression of Ub, GRP78, PDI, CHOP and Bax was increased (P<0.05), while Bcl-2 protein level and Bcl-2/Bax ratio were decreased in model group(P<0.05). Compared with model group, the protein expression of Ub, GRP78, PDI, CHOP and Bax was decreased in SCL (L) group, SCL(M) group and SCL(H) group (P<0.05), while Bcl-2 protein level and Bcl-2/Bax ratio were increased (P<0.05). In control group, neuronal morphology was normal, the protein expression of Bcl-2 was positive and Bax was negative in the cytoplasm. In model group, the neurons were degeneration, the protein expression of Bcl-2 was negative and Bax was positive in the cytoplasm. In SCL (L) group, SCL (M) group and SCL (H) group, the number of degenerative neurons decreased, the protein expression of Bcl-2 was positive and Bax was negative in the cytoplasm. CONCLUSION: SCL inhibit D-galactose-induced brain aging in mice by attenuating apoptosis of endoplasmic reticulum pathway.