The ha72 Core Gene of Baculovirus Is Essential for Budded Virus Production and Occlusion-Derived Virus Embedding,and Amino Acid K22 Plays an Important Role in Its Function

ha72 of Helicoverpa armigera nucleopolyhedrovirus (a homologue of ac78) was identified as a conserved late baculovirus gene and characterized. HA72 localizes in the intranuclear ring zone. By generating mutants, we showed that HA72 is essential for budded virus (BD) production and occlusion-derived virus (ODV) embedding. HA72 also interacted with P33, a baculoviral sulfhydryl oxidase. A point mutation of amino acid 22 from lysine to glutamic acid curtailed BV production and precluded ODV occlusion as well as interaction with P33.     

分子标记技术的发展及应用

介绍了几种应用前景较广的分子标记,如基于DNA杂交技术的分子标记:限制性片段长度多态性(RFLP)和DNA可变串联重复数标记(VNRT);基于PCR技术的分子标记:随机扩增多态性 DNA(RAPD)、酶切扩增多态性(CAPS)、扩增片段长度多态性(AFLP)、微卫星DNA(SSR)和DNA单链构象多态性(SSCP);以及新兴的第3代分子标记,即基于DNA芯片技术的分子标记:单核苷酸多态性(SNP)等。分别阐述了它们的原理、方法步骤与优缺点、应用注意事项和适用范围,同时概述了它们在生物学研究中的应用和进展。     

A nanoprobe for nonprotein thiols based on assembling of QDs and 4-amino-2,2,6,6-tetramethylpiperidine oxide

A new fluorescent nanoprobe, 4-amino-2,2,6,6-tetramethylpiperidine oxide (AT)-functionalized CdTe quantum dots (QDs-AT), was synthesized, for selective detection of nonprotein thiols based on electron transfer (ET). In the presence of nonprotein thiols, the nitroxide radicals in QDs-AT were converted to hydroxylamines, resulting in the fluorescence recovery of the quenched QDs. The detection mechanism of the probe was investigated using Rh-Se-2 probe. The nanoprobe has high sensitivity toward glutathione (GSH) with a detection limit of 7.1 × 10?? M. The fluorescent imaging of living cells showed that QDs-AT could distinguish the concentration differences of GSH in HL-7702 and HepG2 cells.     

斗米虫蛋白体外抗肿瘤活性及其对小鼠巨噬细胞免疫调节作用

该文主要为了研究斗米虫蛋白的体外抗肿瘤活性及其免疫调节作用。提取斗米虫总蛋白,逐级盐析分为3个部分,透析除盐后得到不同蛋白部位,并采用SDS-PAGE检测斗米虫不同蛋白部位的分子量;利用MTT法、流式细胞术等方法研究斗米虫蛋白对人胃癌细胞MFC和小鼠乳腺癌细胞4T1增殖、迁移和凋亡的作用。MTT法研究斗米虫蛋白对小鼠单核巨噬细胞RAW264.7和人脐静脉内皮细胞HUEVC增殖的影响;荧光微球吞噬实验检测斗米虫蛋白对RAW264.7细胞吞噬能力的影响;Griess法检测对RAW264.7细胞释放NO能力的影响;ELISA法检测对RAW264.7细胞的IL-6、TNF-α和IL-1β分泌量;RT-PCR法检测不同浓度的斗米虫蛋白作用后RAW264.7细胞中TNF-α、IL-1、TLR4、MIR-7、IFN-γ、TRL-4、IL-6以及4T1细胞中MMP2、MMP9、STAT3、c-Myc和Sdf1 mRNA水平变化。结果显示,斗米虫蛋白主要分子量集中于63 kDa,斗米虫蛋白对人胃癌细胞MFC及小鼠乳腺癌细胞4T1的增殖表现出较好抑制作用,并呈现出一定剂量依赖关系(P<0.05),对HUEVC细胞没有细胞毒性,对RAW264.7细胞表现出较好的促进增殖的作用(P<0.05)。斗米虫蛋白实验组与正常组细胞相比可以抑制4T1细胞的迁移(P<0.01),可诱导MFC和4T1细胞凋亡(P<0.05);斗米虫蛋白能够提高RAW264.7细胞的吞噬活性、NO释放量、TNF-α、1L-1β和IL-6分泌量及TNF-α、IL-1、TLR4、MIR-7、IFN-γ和IL-6细胞因子的mRNA水平以及能显著下调4T1细胞中MMP2、MMP9、STAT3、c-Myc和Sdf1 mRNA水平(P<0.05,P<0.01)。由此推论,斗米虫蛋白具有较好的体外抗肿瘤活性并且具有潜在的免疫调节作用。     

1000 h Operational Lifetime Perovskite Solar Cells by Ambient Melting Encapsulation

Improving device lifetime is one of the critical challenges for the practical use of metal halide perovskite solar cells (PSCs), wherein a reliable encapsulation is indispensable. Herein, based on an in‐depth understanding of the degradation mechanism for the PSCs, a solvent‐free and low‐temperature melting encapsulation technique, by employing low‐cost paraffin as the encapsulant that is compatible with perovskite absorbers, is demonstrated. The encapsulation strategy enables the full encapsulating operations to be undertaken under an ambient environment. It is found that the strategy not only removes residual oxygen and moisture to prevent the perovskite from phase segregation, but also suppresses the species volatilization to impede absorber decomposition, enabling a PSC devices with good thermal and moisture stability. As a result, the as‐encapsulated PSCs achieve a 1000 h operational lifetime for the encapsulated device at continuous maximum power point output under an ambient environment. This work paves the way for scalable and robust encapsulation strategy feasible to hybrid perovskite optoelectronics in an economic manner.     

Circ_FBLN1 promotes the proliferation and osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by regulating let-7i-5p/FZD4 axis and Wnt/β-catenin pathway

Background

Recently, more and more circular RNAs (circRNAs) have been identified in osteogenesis. In this study, we aimed to explore the effect of circ_FBLN1 on the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs).

Methods

The protein levels of osteogenesis-related genes, let-7i-5p, frizzled class receptor 4 (FZD4), Ki67, Wnt6 and β-catenin were measured by western blot assay. The levels of circ_FBLN1, FBLN1 mRNA and FZD4 mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR) assay. The feature of circ_FBLN1 was investigated by RNase R and Actinomycin D assays. Cell proliferation ability was evaluated by colony formation assay and 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The targeting relationship between let-7i-5p and circ_FBLN1 or FZD4 was verified by dual-luciferase reporter assay.

Results

Circ_FBLN1 level was enhanced during the osteogenic differentiation of hBMSCs. Silencing of circ_FBLN1 repressed cell proliferation and osteogenic differentiation in hBMSCs. For mechanism analysis, circ_FBLN1 was found to act as a sponge for let-7i-5p and FZD4 served as a direct target gene of let-7i-5p. Let-7i-5p was downregulated during the osteogenic differentiation of hBMSCs and let-7i-5p inhibition restored the effects of circ_FBLN1 knockdown on the proliferation and osteogenesis of hBMSCs. Moreover, let-7i-5p overexpression suppressed cell proliferation and osteogenesis in hBMSCs through targeting FZD4. In addition, circ_FBLN1 knockdown reduced the levels of Wnt6 and β-catenin in hBMSCs, indicating the inactivation of Wnt/β-catenin pathway.

Conclusion

Knockdown of circ_FBLN1 inhibited the proliferation and osteogenesis of hBMSCs by regulating let-7i-5p/FZD4 axis and repressing Wnt/β-catenin pathway.

     

六种湿地植物根际氧化还原电位的日变化

在野外条件下,研究人工湿地植物根际氧化还原电位(ORP)随时间的变化及其与主要环境因子的关系。研究了美人蕉(Canna indica Linn.)、风车草(Cyperus flabelliformis Rottb.)、芦苇(Phragmites australis Trin.ex Steud.)、水鬼蕉(Hymenocallis littoralis(Jack.)Salisb.)、紫芋(Colocasia tonoimo Nakai.)和鸢尾(Iris tectorum Maxim.)6种植物在潜流人工湿地中的根际ORP及其日变化。6种湿地植物的根际ORP日变化曲线相似,均为双峰型,双峰值出现在11:00—14:00之间,最大值出现在14:00。各植物的根际ORP日变化基本在130—350 m V之间,以水鬼蕉的变幅最大,风车草和芦苇的变幅较小。不同植物的根际ORP有较大差异,风车草和紫芋的日平均值最大,显著高于鸢尾、美人蕉和水鬼蕉(P0.05);芦苇显著高于鸢尾和美人蕉(P0.05)。ORP与光照强度和气温呈正相关,尤与气温的正相关最为显著。ORP日平均值与植物生物量有显著的正相关性,尤与地下部分生物量相关性最显著。结果表明,人工湿地植物根际ORP因不同植物、一天中不同的时间有较大差异,后者与光照和气温等环境因子密切相关。     

Arsenic Sulfide Promotes Apoptosis in Retinoid Acid Resistant Human Acute Promyelocytic Leukemic NB4-R1 Cells through Downregulation of SET Protein

Tetra-arsenic tetra-sulfide (As₄S₄) is an arsenic compound with anti-tumor activity, especially in acute promyelocytic leukemia (APL) that are resistant to retinoic acid (RA). Although recent studies revealed that the therapeutic action of As₄S₄ is closely associated with the induction of cellular apoptosis, the exact molecular mechanism of action of As₄S₄ in RA-resistant APL remains to be clarified. In this study, we found that As₄S₄-induced apoptosis was accompanied by reduced mRNA and protein expression of SET gene in RA-resistant NB4-R1 cells. Moreover, RNAi knockdown of SET gene further promoted As₄S₄-induced apoptosis, while SET over-expression inhibited it, suggesting that As₄S₄ induces apoptosis through the reduction of SET protein in NB4-R1 cells. We also demonstrated that the knockdown of SET gene resulted in the upregulation of protein phosphatase 2 (PP2A) expression and the downregulation of promyelocytic leukemia and retinoic acid receptor α fusion gene (PML-RARα) expression, which were enhanced by As₄S₄ treatments. By contrast, over-expression of SET gene resulted in PP2A downregulation and PML-RARα upregulation, which were abolished by As₄S₄ pretreatment. Since PP2A is a pro-apoptotic factor and PMLRARα is an anti-apoptotic factor, our results suggest that As₄S₄-induced apoptosis in NB4-R1 cells is through the downregulation of SET protein expression, which in turn increases PP2A and reduces PML-RARα expressions to lead to cell apoptosis.