CHO细胞表达系统研究新进展

中国仓鼠卵巢细胞(ChineseHamsterOvaryCell)是目前重组糖基蛋白生产的首选体系。本文综述近几年来该系统在生产基因工程药物方面的研究进展、存在问题和发展方向。     

Three RNA Binding Proteins Form a Complex to Promote Differentiation of Germline Stem Cell Lineage in Drosophila

In regenerative tissues, one of the strategies to protect stem cells from genetic aberrations, potentially caused by frequent cell division, is to transiently expand the stem cell daughters before further differentiation. However, failure to exit the transit amplification may lead to overgrowth, and the molecular mechanism governing this regulation remains vague. In a Drosophila mutagenesis screen for factors involved in the regulation of germline stem cell (GSC) lineage, we isolated a mutation in the gene CG32364, which encodes a putative RNA-binding protein (RBP) and is designated as tumorous testis (tut). In tut mutant, spermatogonia fail to differentiate and over-amplify, a phenotype similar to that in mei-P26 mutant. Mei-P26 is a TRIM-NHL tumor suppressor homolog required for the differentiation of GSC lineage. We found that Tut binds preferentially a long isoform of mei-P26 3′UTR, and is essential for the translational repression of mei-P26 reporter. Bam and Bgcn are both RBPs that have also been shown to repress mei-P26 expression. Our genetic analyses indicate that tut, bam, or bgcn is required to repress mei-P26 and to promote the differentiation of GSCs. Biochemically, we demonstrate that Tut, Bam, and Bgcn can form a physical complex in which Bam holds Tut on its N-terminus and Bgcn on its C-terminus. Our in vivo and in vitro evidence illustrate that Tut acts with Bam, Bgcn to accurately coordinate proliferation and differentiation in Drosophila germline stem cell lineage.     

灌浆期高温对冬小麦籽粒氨基酸含量和组成的影响

以强筋小麦品种郑麦366和中筋小麦品种洛旱2号为材料,采用盆栽和人工气候室模拟的方法,研究了花后不同时段(灌浆前期即花后5 d,灌浆中期即花后15 d)和持续时间(处理2 d和4 d)的高温胁迫对籽粒氨基酸含量和积累量(以单粒中氨基酸含量表示)的影响。结果显示:(1)高温胁迫显著增加了小麦籽粒中必需氨基酸(EAA)、非必需氨基酸(NAA)和总氨基酸(TAA)含量,但降低了其积累量和EAA/TAA,2品种表现一致;(2)灌浆前期高温胁迫对2品种籽粒氨基酸含量的影响大于灌浆中期,而对氨基酸积累量的影响则相反;(3)高温持续时间对籽粒氨基酸含量的影响在2品种间存在差异,洛旱2号籽粒赖氨酸、EAA、NAA和TAA含量均随高温持续时间的延长显著增加,而郑麦366籽粒中上述指标仅在高温胁迫4 d下较对照增加显著;(4)从受高温胁迫的影响看,籽粒EAA/TAA对高温时段更敏感,而氨基酸含量表现为更易受高温持续时间的影响;(5)高温时段与持续时间的互作效应体现在:灌浆前期籽粒氨基酸积累量2品种均以高温胁迫4 d的影响最大,而灌浆中期则均以高温2 d的降幅最大。上述结果表明,小麦籽粒氨基酸及其组分对高温胁迫的响应不仅在品种间存在差异,且受高温时段和持续时间的影响。     

Platelet activating factor induces transient blood‐brain barrier opening to facilitate edaravone penetration into the brain

The blood–brain barrier (BBB) greatly limits the efficacy of many neuroprotective drugs' delivery to the brain, so improving drug penetration through the BBB has been an important focus of research. Here we report that platelet activating factor (PAF) transiently opened BBB and facilitated neuroprotectant edaravone penetration into the brain. Intravenous infusion with PAF induced a transient BBB opening in rats, reflected by increased Evans blue leakage and mild edema formation, which ceased within 6 h. Furthermore, rat regional cerebral blood flow (rCBF) declined acutely during PAF infusion, but recovered slowly. More importantly, this transient BBB opening significantly increased the penetration of edaravone into the brain, evidenced by increased edaravone concentrations in tissue interstitial fluid collected by microdialysis and analyzed by Ultra‐performance liquid chromatograph combined with a hybrid quadrupole time‐of‐flight mass spectrometer (UPLC‐MS/MS). Similarly, incubation of rat brain microvessel endothelial cells monolayer with 1 μM PAF for 1 h significantly increased monolayer permeability to ¹²⁵I‐albumin, which recovered 1 h after PAF elimination. However, PAF incubation with rat brain microvessel endothelial cells for 1 h did not cause detectable cytotoxicity, and did not regulate intercellular adhesion molecule‐1, matrix‐metalloproteinase‐9 and P‐glycoprotein expression. In conclusion, PAF could induce transient and reversible BBB opening through abrupt rCBF decline, which significantly improved edaravone penetration into the brain.

     

人工红树林湿地系统净化污水研究进展

咸淡水交接处这种特殊的生境造就了红树植物“抗污”和“降污”的功能,但能否将自然红树林湿地作为污水处理场所目前仍有争议。按照构建的人工红树林湿地的类型,分为三大类：模拟潮汐湿地系统、人工红树林污水处理系统以及红树林种植-养殖生态耦合系统。第一类系统模拟了自然海滩上的潮涨潮落的情形,第二类系统与一般人工湿地相类似,而第三类系统是指在养殖塘种植红树植物以去除有机物和营养盐。不同的类型反映了该领域研究工作的不断深入,展现了红树林湿地净化污水从理论探索向实际应用不断靠近的过程。应用人工红树林湿地净化污水具有较大的可行性,前景可观。     

长期施肥对双季稻田甲烷排放和关键功能微生物的影响

研究不同施肥措施对双季稻田甲烷(CH_4)排放特征的影响及其微生物学机理,对合理利用及评价不同施肥模式对水稻生长的影响具有重要意义。以长期施肥定位试验田为平台,采用静态箱-气相色谱法对施用化肥(MF:mineral fertilizer alone)、秸秆还田配施化肥(RF:rice residues plus mineral fertilizer)、30%有机肥配施70%化肥(LOM:30%organic matter plus 70%mineral fertilizer)、60%有机肥配施40%化肥(HOM:60%organic matter plus 40%mineral fertilizer)和无肥(CK:without fertilizer)条件下双季稻田CH_4排放及其微生物学机理进行了分析。结果表明,早稻和晚稻生长期,不同施肥处理稻田CH_4排放通量均显著高于CK,表现为HOMLOMRFMFCK。各处理间CH_4总排放量差异达显著水平,其大小顺序与排放通量趋势一致,以HOM处理为最高,比CK处理增加105.56%,其次是LOM和RF处理,分别比CK处理增加72.97%和54.17%。关键功能土壤微生物测定结果表明,早稻和晚稻各个主要生育时期,各处理稻田土壤产甲烷古菌的数量变化范围为(3.18—81.07)×10~3cfu/g,土壤甲烷氧化细菌的数量变化范围为(24.82—379.72)×10~3cfu/g。稻田土壤产甲烷古菌和甲烷氧化细菌数量大小顺序为HOMLOMRFMFCK,各施肥处理均显著高于CK;HOM、LOM、RF处理显著高于MF、CK处理。双季稻田CH_4排放与稻田土壤产甲烷古菌、甲烷氧化细菌数量变化关系密切。采用有机无机肥配施促进了双季稻田生态系统CH_4的排放和关键功能微生物的数量。