栉孔扇贝和虾夷扇贝杂交子代的GISH鉴定及其免疫学特性

以栉孔扇贝[Chlamys farreri（Jones et Preston）]（♀）和虾夷扇贝（Patinopecten yessoensis）（♂）杂交子代担轮幼虫为材料，分别用栉孔扇贝和虾夷扇贝基因组作探针，采用基因组荧光原位杂交（GISH）的方法，对杂交后代杂交子的确切身份进行初步鉴定。结果表明，子代分别继承了双亲各一套染色体（n=19），为真正的杂交种。为了解杂交扇贝的免疫学特性，在自然海域栉孔扇贝大规模死亡的情况下，分别对杂交扇贝及其亲本3个扇贝群体血细胞的胞内活性氧含量（ROIs）、血清凝集素效价（HA）、溶菌酶活力（LSZ）、抑菌活力、酚氧化酶活性（P0）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）以及酸性磷酸酶（ACP）和碱性磷酸酶（ALP）等9种非特异性免疫学指标进行测定。结果表明，栉孔扇贝除ROIs、SOD、ACP等3个指标显著低于虾夷扇贝外（P〈0．01），其他6种指标均高于虾夷扇贝，且除血清凝集素效价外均达显著水平（P〈0．05）。在杂交子代中，上述免疫指标除SOD活性低于低值亲本外（P〉0．6），其余8种免疫指标均介于双亲之间。杂交子代在9种免疫指标中有8种与母本无显著性差异，而子代与父本之间9种免疫指标中有7种达显著差异（P〈0．05）。这些结果说明，杂交扇贝在非特异性免疫上存在明显的偏母性特征，这点与子代在外形特征上的偏母性相吻合。因此杂交扇贝相对于其母本在生产实践中表现出的一定程度的抗逆优势可能与非特异性免疫无明显关系。[中国水产科学，2006，13（4）：597—602]     

栉孔扇贝异精雌核发育四倍体早期胚胎发育的细胞学荧光显微观察

用1500μW/cm2的紫外线照射长牡蛎(Crassostrea gigas)精子60 s以进行灭活处理,并使之与栉孔扇贝(Chlamys farreri)卵子受精,在卵子受精后排出第一极体前用6-DMAP(50 mg/L)处理受精卵,持续处理35 min,抑制第一极体和第二极体的排放,诱导异精雌核发育四倍体。采用二脒基苯基吲哚(DAPI)荧光染色显微观察法,对灭活的长牡蛎精子诱导的栉孔扇贝雌核发育四倍体早期胚胎发育过程进行细胞学观察。结果表明:经紫外线灭活过的长牡蛎精子进入栉孔扇贝卵子后发生轻微膨胀;在第一次卵裂中期,精核形成一致密的染色质小体(DCB),位于两组分开的母本染色体之间,不参与核分裂;第一次卵裂结束时DCB滞留于两卵裂球的分裂沟上或进入其中一分裂球中;第二次卵裂过程中,DCB的去向与第一次卵裂时基本一致。6-DMAP处理有效地抑制了第一极体和第二极体的排出,从而使雌核四倍化。对担轮幼虫染色体倍性分析结果表明,通过本方法可以获得6.25%的四倍体幼虫。本研究还对灭活的异源长牡蛎精子诱导栉孔扇贝雌核发育四倍体过程中产生的复杂倍性、核物质分离紊乱及多精附卵现象进行了观察和分析。     

张河湾抽水蓄能电站1号机组转子安装出现的问题及解决方法

文章介绍了张河湾抽水蓄能电站转子结构特点及安装工艺,分析了转子安装过程中出现的磁轭冲片受损、拉紧度不够等原因及解决办法,介绍了如何解决安装过程中分块式压板的水平度调整。     

肉桂醛、柠檬醛对犬白色念珠菌黏附力及超微结构的影响

根据先前的研究,肉桂醛和柠檬醛都有较好的抗真菌活性。本研究对肉桂醛、柠檬醛在真菌黏附口腔黏膜上皮以及对犬白色念珠菌超微结构的影响进行探索。结果表明,药物处理组白色念珠菌黏附率显著下降。扫描电镜显示,经过药物处理的犬白色念珠菌细胞表面粗糙、皱缩、萎陷、外膜空洞、胞浆内容物流出。透射电镜下,细胞普遍发生细胞壁增厚,质壁分离,萎陷,胞浆内容物外流等损伤。     

Effects of notch1 gene on proliferation and cell cycle of human glioma U251 cells

AIM: To investigate the effect of notch1 gene on the change of proliferation and cell cycle in human glioma U251 cell line. METHODS: The lentiviral vectors, which express notch1 shRNA or notch1 intracellular domain (NICD), were constructed and transfected into U251 cells, respectively. RT-PCR and Western blotting were applied to monitor the validity of down-regulation of notch1 expression and over-expression of NICD. MTT assay was performed to examine the cell proliferation. Flow cytometric analysis was used to detect the cell cycle. RESULTS: The lentiviral vectors, which expressed notch1 shRNA and NICD, were efficient in silencing notch1 expression and over-expression of NICD. Down-regulation of notch1 gene by RNAi inhibited the cell proliferation remarkably (P<0.01), arrested cell cycle at G1 phase (P<0.01) and decreased the cell number of S phase (P<0.01). Over-expression of NICD enhanced the cell proliferation significantly (P<0.01), promoted the cell cycle at G1 phase (P<0.05) and increased the cell number of S phase (P<0.01). CONCLUSION: notch1 gene, which leads to change the proliferation and cell cycle in human glioma U251 cell line, is likely to be potential molecular target for glioma in gene therapy.     

Effects of Ginkgolide B on glutamate-induced apoptosis and calcium overload in cultured rat cortical neurons

AIM: To observe the effects of ginkgolide B on the neuron apoptosis induced by glutamate and explore whether this effects are related to the changes of calcium in neurons. METHODS: Primary neuron culture was prepared according to a previously reported procedure with slight modification. Neuron damage was induced by 0.8 mmol/L glutamate. The cell survival rate was examined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. The neuron morphological changes, Hoechst 33258 unclear-staining analysis and agarose gel electrophoresis of DNA were measured as the indexes of cell apoptosis. Intracellular free Ca2+ concentration in neurons was measured by using the fluorescent Ca2+ indicator Fura-2/AM. RESULTS: The cells, exposed to glutamate (0.8 mmol/L), showed characteristic change of apoptosis and calcium overload, which were relieved by the treatment of ginkgolide B (10-250 μmol/L), with survival increasing and cell morphology restoring and DNA fragment decreasing. CONCLUSIONS: Ginkgolide B prevents the neurons from glutamate neurotoxicity by inhibiting glutamate-induced apoptosis. The potential mechanism of its action may be related to the competitive PAF receptor binding of ginkgolide B and decreasing the intracellular calcium concentration in neurons.     

安哥拉山羊与建昌黑山羊及其杂种后代的DNA指纹分析

以 (CA/GATA/TCC) 5探针 ,HinfI酶切 ,研究安哥拉山羊和建昌黑山羊及安哥拉山羊与建昌黑山羊级进杂交的F2 、F3的DNA指纹图谱。结果 ,在供试山羊中 ,个体平均检出 1 8.2± 0 .4条谱带 ,安哥拉山羊、F2 、F3的鉴别机率分别为 1 .38× 1 0 - 12 ,1 .1 8× 1 0 - 13,0 .50× 1 0 - 10 ;群体内相似系数安哥拉山羊为0 .4 539,F2 为 0 .4 0 77,F3为 0 .61 1 1 ;亲子鉴定的父权概率W =0 .9889;安哥拉山羊与建昌黑山羊比较 ,2 .3kb和 8.6kb谱带为安哥拉山羊的特异谱带     

Effect of TLR4 activation on endothelium adhesion function and atorvastatin intervention

AIM：To investigate the modulation of LOX-1 and monocyte-endothelium adhesion by TLR4 activation in human umbilical vein endothelial cells (HUVECs),and explore LOX-1’s role in mediating monocyte-endothelium adhesion,and the effect of atorvastatin.METHODS：TLR4 and LOX-1 mRNA were measured by RT-PCR.The expression percentage of TLR4 and LOX-1 positive cells were detected by flow cytometry.The adhesive percentage between monocytes and HUVECs were determined by counting.RESULTS：Incubation by LPS (1 mg/L) for 24 hours upregulated TLR4,LOX-1 mRNA and protein expression in HUVECs，and increased the percentage of monocyte adhesion to endothelium.Pretreatment of cell with anti-LOX-1 partly abolished the increase of monocyte adhesion to endothelium.Atorvastatin (10 μmol/L) inhibited LPS-mediated effects above.CONCLUSION：TLR4 activation upregulates LOX-1 expression and increases monocyte-endothelium adhesion.LOX-1 partly involves in LPS-induced monocyte-endothelium adhesion,atorvastatin may have protective effects on endothelium by inhibiting TLR4 and TLR4-induced LOX-1 expression.     

Increased expression and possible roles of nicotinamide N-methyltransferase in pancreatic islets of STZ-induced diabetic monkeys

AIM: To verify and localize the expression of nicotinamide N-methyltransferase (NNMT) in pancreas of streptozotocin(STZ)-induced diabetic monkeys and understand its important role in β-cell destruction in the pathogenesis of diabetes. METHODS: Through an olig-microarray gene chip, NNMT was identified as the most obviously up-regulated genes in pancreas of STZ-induced diabetic monkeys versus controls. Semiquantitative RT-PCR and Western blotting were performed to verify the differential expression at mRNA and protein level respectively. Then the cellular localization of NNMT expression within pancreas was identified by immunohistochemical(IHC) staining.RESULTS: An obvious high expression of NNMT at both mRNA and protein levels was shown in pancreas of STZ-induced diabetic monkeys compared to that of controls. Further localization of the protein by IHC staining in pancreas specimens showed that its altered expression was restricted to central islets, most of which were β cells.CONCLUSION: Expression of NNMT is increased in islets of STZ- induced diabetic monkeys, which infers that NNMT might participate in the process of β cell damage in diabetes probably through the mechanism of energy metabolism disturbance.