从烟草悬浮细胞中分离液泡前体的方法研究

通过跟踪含有液泡前体特异标记物的组分, 利用细胞壁酶水解胞壁制造原生质体, 并用真针头挤压的方法裂解细胞, 用蔗糖密度梯度离心的方法分离液泡前体. 通过相差显微镜观察, 发现所分离的液泡前体保持完整的结构. 利用不同细胞器的标记物抗体进行免疫标记检测, 证明所纯化的液泡前体不含其他细胞器, 具有较高的纯度.     

锚索锚杆联合支护在破碎顶板中的应用

结合工程工作面概况，就锚索和锚杆补强加固支护参数的选择进行了介绍，并指出锚索、锚杆的联合支护，保证了顶板的完整性，有效地控制了巷道的变形。     

Altered mRNA and Protein Expression of Monocarboxylate Transporter MCT1 in the Cerebral Cortex and Cerebellum of Prion Protein Knockout Mice

The effect of a cellular prion protein (PrP^c) deficiency on neuroenergetics was primarily analyzed via surveying the expression of genes specifically involved in lactate/pyruvate metabolism, such as monocarboxylate transporters (MCT1, MCT2, MCT4). The aim of the present study was to elucidate a potential involvement of PrP^c in the regulation of energy metabolism in different brain regions. By using quantitative real-time polymerase chain reaction (qRT-PCR), we observed a marked reduction in MCT1 mRNA expression in the cortex of symptomatic Zürich I Prnp^−/− mice, as compared to their wild-type (WT) counterparts. MCT1 downregulation in the cortex was accompanied with significantly decreased expression of the MCT1 functional interplayer, the Na⁺/K⁺ ATPase α2 subunit. Conversely, the MCT1 mRNA level was significantly raised in the cerebellum of Prnp^−/− vs. WT control group, without a substantial change in the Na⁺/K⁺ ATPase α2 subunit expression. To validate the observed mRNA findings, we confirmed the observed change in MCT1 mRNA expression level in the cortex at the protein level. MCT4, highly expressed in tissues that rely on glycolysis as an energy source, exhibited a significant reduction in the hippocampus of Prnp^−/− vs. WT mice. The present study demonstrates that a lack of PrP^c leads to altered MCT1 and MCT4 mRNA/protein expression in different brain regions of Prnp^−/− vs. WT mice. Our findings provide evidence that PrP^c might affect the monocarboxylate intercellular transport, which needs to be confirmed in further studies.     

链霉亲和素-蛋白A融合蛋白基因在大肠杆菌中的表达

将合链霉亲和亲-蛋白A（Streptavidin－proteinA）融合蛋白基因的表达质粒pTSAPA导入大肠杆菌BL21（DE3），成功地表达了该融合蛋白，经SIJS－PAGE及免疫印迹等方法证实该蛋白既具有IgG的结合活性，又具生物素结合活性。     

Anti-inflammatory effects and mechanisms of usnic acid

The anti-inflammatory effect and mechanism of Usnic acid (UA) were explored on lipopoly-saccharide (LPS)-stimulated RAW264.7 cell line.The effects of UA on pro-inflammatory cytokines including tumor necrosis factor-alfa (TNF-α),interleukin-6 (IL-6) and interleukin-1 beta (IL-1β),pro-inflammatory mediators such as nitric oxide (NO),inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were studied by sandwich ELISA,real-time PCR and western blot analyses.Similarly,the effect of UA on anti-inflammatory cytokine interleukin-10 (IL-10) and anti-inflammatory mediator heme oxygenase-1 (HO-1) were also studied following the same methods.Furthermore,nuclear factor-κB (NF-κB) was assayed by immunocytochemistry.The results showed that UA has anti-inflammatory effect by down-regulatinng iNOS,COX-2,IL-1β,IL-6 and TNF-α,COX-2 gene expression through the suppression of NF-κB activation and increasing anti-inflammatory cytokine IL-10 and anti-inflammatory mediator HO-1 production.     

往复式压缩机连杆的失效原因

某企业乙二醇回收气往复式压缩机连杆大头盖及其紧固螺栓发生了断裂事故。对该连杆大头盖和紧固螺栓进行了断口宏观形貌、材料化学成分、扫描电镜、显微组织等的检验分析。结果表明:造成此次压缩机事故的主要原因是压缩机连杆螺栓预紧力不足导致连杆两瓦之间存在间隙,造成该部位螺栓表面产生微动磨损诱发裂纹源,产生疲劳断裂,进而导致连杆大头盖断裂、连杆发生弯曲变形。     

Calmodulin and Its Interactive Proteins Participate in Regulating the Explosive Growth of Alexandrium pacificum (Dinoflagellate)

Alexandrium pacificum is a typical dinoflagellate that can cause harmful algal blooms, resulting in negative impacts on ecology and human health. The calcium (Ca²⁺) signal transduction pathway plays an important role in cell proliferation. Calmodulin (CaM) and CaM-related proteins are the main cellular Ca²⁺ sensors, and can act as an intermediate in the Ca²⁺ signal transduction pathway. In this study, the proteins that interacted with CaM of A. pacificum were screened by two-dimensional electrophoresis analysis and far western blots under different growth conditions including lag phase and high phosphorus and manganese induced log phase (HPM). The interactive proteins were then identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Four proteins were identified, including Ca²⁺/CaM-dependent protein kinase, serine/threonine kinase, annexin, and inositol-3-phosphate synthase, which all showed high expression levels under HPM. The gene expression levels encoding these four proteins were also up-regulated under HPM, as revealed by quantitative polymerase chain reaction, suggesting that the identified proteins participate in the Ca²⁺ transport channel and cell cycle regulation to promote cell division. A network of proteins interacting with CaM and their target proteins involved in the regulation of cell proliferation was raised, which provided new insights into the mechanisms behind the explosive growth of A. pacificum.     

Auto-microfluidic thin-film chip for genetically modified maize detection

As a thin-film chip method, reverse dot blot hybridization (RDBH) has been employed to detect hazardous substances, but an automatic RDBH instrument with low workload, high accuracy and stability is still urgently needed. This paper presents our newly-developed auto-microfluidic thin-film chip (AMTC) method for multiplex screening of genetically modified (GM) maize. With specific DNA probes for genetically modified (GM) maize being immobilized on a square nylon thin-film, it was placed into a micro-reaction cell of the AMTC device. Then biotin-labeled PCR products with target DNA fragments for template amplification were added to the micro-reaction cell using a microfluidic system. When the PCR products passed the square nylon thin-film, the target DNA fragments were captured by the complementary action of DNA, where the signal was visualized with streptavidin link-coupled alkaline phosphatase color development kit. The sensitivity of GM maize detection reached 0.1% quality percentage and its stability and consistency could satisfy the requirements for practical applications. Performance advantages of the ATMC are manifold, being embodied in aspects such as easy and straightforward operation, low costs and less workload.