The Potential Role of the Methionine Aminopeptidase Gene PxMetAP1 in a Cosmopolitan Pest for Bacillus thuringiensis Toxin Tolerance

Methionine aminopeptidases (MetAPs) catalyze the cleavage of the N-terminal initiator methionine (iMet) in new peptide chains and arylamides, which is essential for protein and peptide synthesis. MetAP is differentially expressed in two diamondback moth (DBM; Plutella xylostella) strains: the G88 susceptible strain and the Cry1S1000 strain, which are resistant to the Bt toxin Cry1Ac, implicating that MetAP expression might be associated with Bt resistance. In this study, we identified and cloned a MetAP gene from DBMs, named PxMetAP1, which has a CDS of 1140 bp and encodes a 379 amino acid protein. The relative expression of PxMetAP1 was found to be ~2.2-fold lower in the Cry1S1000 strain compared to that in the G88 strain. PxMetAP1 presents a stage- and tissue-specific expression pattern, with higher levels in the eggs, adults, integument, and fatbody of DBMs. The linkage between PxMetAP1 and Cry1Ac resistance is verified by genetic linkage analysis. The knockout of PxMetAP1 in G88 by CRISPR/Cas9 leads to a ~5.6-fold decrease in sensitivity to the Cry1Ac toxin, further supporting the association between the PxMetAP1 gene and Bt tolerance. Our research sheds light on the role of MetAP genes in the development of Bt tolerance in P. xylostella and enriches the knowledge for the management of such a cosmopolitan pest.     

From 2D to 3D Co-Culture Systems: A Review of Co-Culture Models to Study the Neural Cells Interaction

The central nervous system (CNS) controls and regulates the functional activities of the organ systems and maintains the unity between the body and the external environment. The advent of co-culture systems has made it possible to elucidate the interactions between neural cells in vitro and to reproduce complex neural circuits. Here, we classified the co-culture system as a two-dimensional (2D) co-culture system, a cell-based three-dimensional (3D) co-culture system, a tissue slice-based 3D co-culture system, an organoid-based 3D co-culture system, and a microfluidic platform-based 3D co-culture system. We provide an overview of these different co-culture models and their applications in the study of neural cell interaction. The application of co-culture systems in virus-infected CNS disease models is also discussed here. Finally, the direction of the co-culture system in future research is prospected.     

Intelligent Drug Delivery by Peptide-Based Dual-Function Micelles

To endow the polymeric prodrug with smart properties through a safe and simple method, matrix metalloproteinase (MMPs) responsive peptide GPLGVRGDG was introduced into the block copolymer to prepare TPGS₃₃₅₀-GPLGVRGDG-DOX&DOX micelles, where TPGS₃₃₅₀ is D-α-tocopheryl polyethylene glycol 3350 succinate. During the doxorubicin delivery, the cleavage of the peptide chain triggers de-PEGylation, and the remaining VRGDG sequence was retained on the surface of the micelles, which can act as a ligand to facilitate cell uptake. Moreover, the cytotoxicity of TPGS₃₃₅₀-GPLGVRGDG-DOX&DOX micelles against 4T1 cells was significantly improved, compared with TPGS₃₃₅₀-GPLGVRG-DOX&DOX micelles and TPGS₃₃₅₀-DOX&DOX micelles. During in vivo studies, TPGS₃₃₅₀-GPLGVRGDG-DOX&DOX micelles exhibited good anticancer efficacy with long circulation in the body and more efficient accumulation at the tumor site. Therefore, TPGS₃₃₅₀-GPLGVRGDG-DOX&DOX micelles have improved antitumor activity and reduced toxic side effects. This work opens new potential for exploring the strategy of drug delivery in clinical applications.     

Precision Killing of Sinoporphyrin Sodium-Mediated Photodynamic Therapy against Malignant Tumor Cells

Photodynamic therapy (PDT) has significant advantages in the treatment of malignant tumors, such as high efficiency, minimal invasion and less side effects, and it can preserve the integrity and quality of the organs. The power density, irradiation time and photosensitizer (PS) concentration are three main parameters that play important roles in killing tumor cells. However, until now, the underlying relationships among them for PDT outcomes have been unclear. In this study, human malignant glioblastoma U-118MG and melanoma A375 cells were selected, and the product of the power density, irradiation time and PS concentration was defined as the total photodynamic parameter (TPP), in order to investigate the mechanisms of PS sinoporphyrin sodium (DVDMS)-mediated PDT (DVDMS-PDT). The results showed that the survival rates of the U-118MG and A375 cells were negatively correlated with the TPP value in the curve, and the correlation exactly filed an e-exponential function. Moreover, according to the formula, we realized controllable killing effects of the tumor cells by randomly adjusting the three parameters, and we finally verified the accuracy and repeatability of the formula. In conclusion, the establishment and implementation of a newly functional relationship among the PDT parameters are essential for predicting PDT outcomes and providing personalized precise treatment, and they are contributive to the development of PDT dosimetry.     

Identification and Functional Analysis of Key Autophosphorylation Residues of Arabidopsis Senescence Associated Receptor-like Kinase

Reversible protein phosphorylation mediated by protein kinases and phosphatases plays important roles in the regulation of leaf senescence. We previously reported that the senescence-associated leucine-rich repeat receptor-like kinase AtSARK autophosphorylates on both serine/threonine and tyrosine residues and functions as a positive regulator of Arabidopsis leaf senescence; the senescence-suppressed protein phosphatase SSPP interacts with and dephosphorylates the cytoplasmic domain of AtSARK, thereby negatively regulating leaf senescence. Here, 27 autophosphorylation residues of AtSARK were revealed by mass spectrometry analysis, and six of them, including two Ser, two Thr, and two Tyr residues, were further found to be important for the biological functions of AtSARK. All site-directed mutations of these six residues that resulted in decreased autophosphorylation level of AtSARK could significantly inhibit AtSARK-induced leaf senescence. In addition, mutations mimicking the dephosphorylation form of Ser384 (S384A) or the phosphorylation form of Tyr413 (Y413E) substantially reduced the interaction between AtSARK and SSPP. All results suggest that autophosphorylation of AtSARK is essential for its functions in promoting leaf senescence. The possible roles of S384 and Y413 residues in fine-tuning the interaction between AtSARK and SSPP are discussed herein.     

大孔树脂纯化血管紧张素酶抑制肽VLPVPR的工艺优化

考察大孔树脂(AB8,D101,DM130,HPD100B,HPD826)对血管紧张素酶抑制肽VLPVPR的吸附性能及纯化效果,确立纯化VLPVPR的较优工艺。结果表明,HPD100B最适于VLPVPR的纯化。最优纯化工艺为:温度20℃,pH9.8,上样流速为3mL/(cm2·min),上样量达到180mL,洗脱液乙醇溶液浓度为60%,洗脱流速为0.25mL/(cm2·min),洗脱体积为45mL。此条件下,VLPVPR的解吸率达93.1%,纯度为28.8%,血管紧张素酶抑制率IC50为4.1μmol/L。      

河北平泉农田土壤放线菌物种多样性筛查

针对河北平泉县典型的农田土壤类型,采用基于全长16S rDNA序列系统发育分析的免培养技术,并结合放线菌门特异引物的筛查,揭示了土壤中放线菌的物种多样性。构建的放线菌16S rRNA克隆文库中的596个放线菌序列归属于29个属,17个科,12个亚目和2个亚纲,并确定了其优势菌属,其中芽球菌属(Blastococcus)(占放线菌总克隆数的12.75%),类诺卡氏菌属(Nocardioides)(9.39%),节杆菌属(Arthrobacter)(8.72%),小月菌属(Microlunatus)(5.37%),酸土单胞菌属Aciditerrimonas(4.69%),沉积岩杆菌属Ilumatobacter(3.35%)。本文为深入理解放线菌的生态功能,及更深层次的放线菌资源的研究、开发和利用提供宝贵的科学资料和丰富的物种信息资源。      

大豆磷脂酰肌醇的提取工艺研究

为探究溶剂分提法提取较高纯度的磷脂酰肌醇(PI),以提取PC后的大豆粉末磷脂为原料,在单因素实验的基础上,进行响应面优化实验,以PI含量为指标,确定了以碱性乙醇为溶剂提取磷脂酰肌醇的最佳工艺条件:温度为40℃、液料比为20∶1、碱性乙醇氨水浓度为2.60mL 25%氨水/100mL无水乙醇、时间50min、提取两次。在此条件下,磷脂酰肌醇的纯度为58.86%,得率为70.14%。      

基于迭代线性约束最小方差的稳健自适应脉冲压缩方法

针对常规自适应脉冲压缩方法在目标散射点与采样中心失配时旁瓣抑制性能下降的问题,该文提出一种基于迭代线性约束最小方差(RLCMV)的自适应脉冲压缩方法。该方法首先将自适应波束形成器算法引入到自适应脉冲压缩滤波器设计中。其次对目标及干扰单元进行线性约束,并用对角加载技术避免矩阵出现病态。最后构造了迭代运算方法,依次抑制不同大小目标的距离旁瓣。仿真结果表明,该算法可以有效抑制散射点随机分布目标的距离旁瓣,对散射点与采样中心失配情况具有较好的稳健性,在多目标及距离扩展目标场景中达到较好的旁瓣抑制性能,并在一定程度上提高了多普勒容性。